Objective To understand the psychological stress situation of the nursing staffs in a Class A Tertiary Hospital in Changchun City in Jilin Province and analyze its causes， and to provide the scientific basis for improving the psychological stress situation of the nursing staffs and promoting the quality of medical services. Methods The nursing staffs who worked in a Class-A Tertiary Hospital in Changchun City in Jilin province were regarded as the subjects， and the data were collected by Wenjuanxing Network Questionnaire，the Kessler（K10） Scale was used to evaluate the psychological stress levels of the nursing staffs，the Stressor Scale was used to analyze the main stressors， and multivariate Logistic regression analysis was used to analyze the influencing factors of psychological stress levels of the nursing staffs. Results A total of 691 valid questionnaires were collected； the mean K10 score of the nursing staffs was （23.31±8.16） points，and 56.87% of the nursing staffs were under high psychological stress level （K10 score ≥22 points）； there was significant difference in the detection rate of high psychological stress level among the nursing staffs with different ages（P<0.05）. The multivariate Logistic regression analysis results showed that the main risk factors of psychological stress level inccreasing of the nursing staffs were night shift （ OR=1.930， 95% CI： 1.299－2.869，P=0.001） and being attacked or verbally abused by the patients and their family members （OR=3.945， 95% CI： 2.829－5.502，P<0.001）； the total score and the scores on various dimensions of Stressor Scale of the nursing staffs with the high levels of psychological stress were higher than those with the low or average levels of psychological stress（P<0.05）. Conclusion The levels of psychological stress of the nursing staffs in our hospital and the wide range of stressors is wild； the corresponding measures should be put forward according to the workload and doctor-patient contradictions to reduce the psychological stress of the nursing staffs and increase their psychological health levels.

Abstract:Objective
 To study the protective effect of panax quinquefolium 20s-protopanaxtriolsaponins (PQTS) on acute myocardial infarction in rats,and to clarify its mechanism. Methods The acute myocardial infarct model was prepared by left anterior descending  coronary occulusion for 24 h in openchest anesthetized rats. The rats were divided randomly into sham group,ischemia-reperfusion model group and PQTS 12.5,25.0， and 50.0 mg•kg-1 groups. All animals were administered one time everyday with intraperitoneal injection(i.p) for 3 d. The myocardial infarct size (MIS) was calculated. The activities of serum creatine hosphokinase(CK),lactate dehydrogenase (LDH),aspartate aminotransferase (AST) and superoxide dismutase(SOD),and the contents of malondialdehyde(MDA),nitric oxide(NO) and free fatty acid(FFA) were determined. Blood was collected to observe low shearing specific viscosity,middle shearing specific viscosity and high shearing specific viscosity of whole blood and plasma viscosity. At the same time,the platelet aggregation rate was also determined. Results Compared with model group,the MIS  in the  rats treated by PQTS (in  dosages of 12.5,25.0 and 50 mg•kg-1•d-1,i.p×3 d) were significantly reduced(P<0.05);the activities of serum CK,LDH and AST,and the contents of serum FFA and MDA were declined(P<0.05 or P<0.01);the activitie of SOD,and the content of NO were increased markedly(P<0.05). In addition,the low shearing specific viscosity,middle shearing specific viscosity and high shearing specific viscosity of whole blood and plasma viscosity as well as platelet aggregation rates were also declined significantly(P<0.05 or P<0.01). Conclusion PQTS has a significant protective effect on acute myocardial infarction,which may be related to increasing the activity of antioxidase in body,scavenging the damage of peroxidation from oxygen free radicals,correcting metabolic disorder of FFA in myocardial ischemia,and decreasing the viscosity of blood and plasma and function of the platelet aggregation etc.

Abstract:Objective
To observe and compare the photochemotherapeutic effects of BCPD-17 (hematoporphyrin derivative) and PSD-007 on osteosarcoma LM-8 transplanted in mice and to explore a novel photosenitizer for photodynamic therapy of  osteosarcoma Methods The tumor model was made by subcutaneous injection of osteosarcoma LM-8 on C3H mice.When the tumors reached about 6-8 mm in diameter,the tested mice were randomly divided into control group,PSD-007 group (5 mg•kg^-1),BCPD-17 groups (5 and 10 mg•kg^-1).After received intravenous injection of 5 or 10 mg•kg^-1 of PSD-007 and BCPD-17,respectively,the tumors were irradiated vertically with 630 nm or 670 nm laser (power density 200 mW•cm-2,energy density 240 J•cm^-2) for 20 min.The size of osteosarcoma was measured 7 d  after treatment.And all the tumors were extirpated,weighted and taken for pathological examination.The inhibitory rate of tumor was calculated.Results The results of pathohistological observation showed that in control group the size of mouse osteosarcoma was big and hyperchromatic with obvious nuclear atypia;in PDT treatment group,the turmor cells presented apoptosis,vacuolar degeneration and karyopycnosis,especially in BCPD-17 group. Compared with control group,the volumes and the weights of the tumors in PDT treatment groups were obviously decreased,there were significant differences(P<0.01).Compared with PSD-007 group,the inhibitory rates of tumor in BCPD-17 groups irradiated by the laser with wavelength 670 nm were increased,there were significant differences(P<0.01).Conclusion Both of the photosenitizers BCPD-17 and PSD-007 could obviously inhibit the growth of the tumors.Photosenitizer BCPD-17 has more anti-tumor effect.

Abstract:Objective To investigate the inhibitory effects of Juglans Mandshurica Maxim stem-bark extract（JMME） on SMMC-7721，MCF-7 and A549 cells in vitro,and to confirm the antitumor medicative value of JMME, and to verify antitumor mechanisms of JMME by examining the apoptosis and telomerase activitiy change induced by JMME.Methods SMMC-7721， MCF-7 and A549 cells in culture medium were treated with 25,50,100,200,400 and 800 mg•L^-1 JMME respectively.The inhibitory rates of these cells were measured by MTT assay.After SMMC-7721,MCF-7 and A549 cells in culture medium were respectively treated with 200 mg•L^-1 JMME,the morphological changes of cells were observed under inverted microscope and the apoptosis was detected by agarose gel electrophoresis assay.Then PCR-enzyme-linked immunosorbent assay(ELISA) was used to detect telomerase activities of SMMC-7721,MCF-7 and A549 cells.Results The inhibitory rates (%) of JMME with different concentrations（25,50,100,200,400 and 800 mg•L^-1） on SMMC-7721 were 13.06，21.77，29.88，41.74，69.82 and 78.08，respectively； 24.90，35.61，45.42，65.65，69.69 and 81.39 on MCF-7； 12.32，26.21，41.65，55.38，62.87 and 80.13 on A549.The results showed that JMME had cytolytic action on tumor cells compared with control group（P<0.05）.The inhibitory rates increased with the increasing of the concentration of JMME(P<0.05）.IC₅₀ of SMMC-7721,MCF-7 and A549 after treated with JMME were 211.21,111.07 and 176.20 mg•L^-1,respectively.The IC₅₀ of MCF-7 cells were lower than those of SMMC-7721 and A549 cells.The typical apoptosis morphology was identified under inverted microscope.The typical ladder bar was observed.These results demonstrated that the apoptosis of tumor cells were induced by JMME.The telomerase activities of SMMC-7721,MCF-7 and A549 were stronger than that in negative control ，but the telomerase activities of the tumor cells treated with extracts were much weaker than that of the tumor cells untreated.The inhibitory rates were 42.86%,73.30% and 58.78%,respectively.Conclusion JMME has antitumor effect in vitro.It’ s mechanism is associated with inducing tumor cells apoptosis and inhibiting telomerase activity.

Objective To study the effect of Acanthopanax senticosus saponins (ASS) on organ blood flow and vascular resistance in anesthetized dogs. Methods The cerebral blood flow(CBF), cerebral vascular resistance(CVR),peripheral blood flow(PBF),peripheral vascular resistance(PVR), blood pressure(SDP and DBP) and heart rate(HR) were measured using the MF-27 electromagnetic flowmeter. Results ASS (15 and 30 mg•kg^-1 iv perfusion for 60min) augmented CBF and PBF, decre ased CVR and PVR within 180 min and slowed down HR at 60-180 min. In addition, ASS (30 mg•kg^-1 iv perfusion for 60 min) could obviously reduce SBP and DBP . Conclusion Protective effect of ASS on experimental cerebral ischemia may be rela ted to improving cerebral circulation and increasing cerebral blood supplyment.

Objective：Objective To investigate the genes related to lactic acid metabolism in lung adenocarcinoma （LUAD） tissue and establish the prognostic score model based on the genes related to lactic acid metabolism， and to clarify its ability to predict the prognosis of LUAD. Methods The LUAD related lactate metabolism genes were screened by The Cancer Genome Atlas（TCGA） database.Univariate and multivariate Cox regression analysis and LASSO regression analysis were used to obtain the key genes and the lactate metabolism score model for LUAD was constructed，and the predictive ability of the model was verified by Kaplan-Meier survival analysis and receiver operating characteristic（ROC） curve， and the relationships between the model and the clinical characteristics of the patient and the abundances of immune cell infiltration were evaluated by TIMER method. Results Sixteen lactate metabolism genes were successfully screened and the score model was constructed. The survival analysis results showed that the overall survival（OS） of the patients in low-risk group was significantly higher than that in high-risk group （P<0.01）， and the area under ROC curve （AUC） was higher than 0.7；the multivariate Cox regression analysis results showed that 23 lactic acid metabolism genes were the independent prognosis genes for the LUAD patients，which could accurately assess the survival rate of LUAD patients in combination with other clinical features of the patients（P<0.01），and the lactic acid score was negatively correlated with the percentages of B lymphocytes（r=－0.326，P<0.001），CD4 T lymphocytes（r=－0.196，P<0.001），CD8 T lymphocytes（r=－0.094，P=0.036），macrophages（r=－0.198，P<0.001），and dendritic cells（r=－0.119，P=0.008）. Conclusion The lactate metabolism score model can well evaluate the prognosis of LUAD and the state of tumor microenvironment （TIMER）， and can be used as a biomarker to predict the prognosis of LUAD.

Objective To induce and culture purity dendritic cells (DCs) of different maturation phase from human peripheral blood monocytes in vitro. Methods The monocytes were acquired from human peripheral blood mononuclear cells（PBMC）by adhesion method.The first step was to work on a 7-day culture of monocytes in medium supplement with GM-CSF and IL-4, and the cells were actually immature. In the second step,the cells were cultured in the medium supplement with GM-CSF and TNF-α,and cultured until the 14th day. Mature DCs were obtained. Cells harvested were identified for their morphology by microscope, properties of DCs detected by FCM, function with MTT method. Results Immature dendritic cells expressed CD1a molecules and costimulating molecules in middle level, HLA-DR in high level. In maturation phase DCs expressed costimulating molecules, HLA-DR molecules, CD83, and CD25 highly. These DCs could stimulate proliferation of allogenic T lymphocytes and increase killing ability of CTL activated by DCs. Conclusion The method to obtain DCs of different maturation phases from human peripheral blood monocytes is successfully set up. A number of DCs with high purity are obtained.

To analyze the etiology， clinical manifestations and treatment of one patient with type 1 diabetes mellitus（T1DM） complicated with periodontitis，and to clarify the importance of early prevention and treatment of periodontitis associated with T1DM， and to provide the basis for the diagnosis and treatment for the patients and clinicians. Methods The clinical data of a female patient with the history of T1DM for 8 years were collected and the patient visited houspital due to “gingival atrophy for 1 year”. During the treatment， the blood glucose level of patient was monitored for a long time， and the color， morphology and texture of gingiva， probing depth（PD） of periodontal pocket， positive rate of bleeding on probing（BOP），and morphology of the alveolar bone were recorded 6 weeks，3 months，6 months，and 1 year after the initial periodontal therapy. The related literatures were reviewed，and the etiology， clinical characteristics，and the methods of prevention， diagnosis and treatment of the T1DM patient complicated with periodontitis were summarized. Results Before and after initial periodontal therapy， the blood glucose level of the patient was stable without significant fluctuations. Six weeks after initial periodontal therapy， the color of the patients’ gingiva became lighter， the degree of swelling was decreased， the texture was tough， the PD of periodontal pocket was decreased， and the positive rate of BOP was decreased to 13%. One year after initial periodontal therapy， a small amount of tartar and soft dirt could be seen on some tooth surfaces， and the gingiva of the other teeth were light pink， thin and tough.The PD was 0－4 mm， and the BOP percentage was about 14%， and there was no loose teeth in the whole mouth. Conclusion The patients with T1DM complicated with periodontitis should undergo the regular periodontal examinations and strict plaque control. On the premise of stable blood glucose， the initial periodontal therapy can effectively reduce the occurrence and development of the periodontal disease.

Objective To discuss the effect of long non-coding RNA （lncRNA） gastric cancer related transcript 2（GACAT2） gene on the proliferation and stemness of the lung cancer A549 and H1299 cells， and to clarify the effect of lncRNA GACAT2 gene on the occurrence and development of lung cancer. Methods The full-length sequence of lncRNA GACAT2 gene was amplified by real-time fluorescence quantitative PCR （RT-qPCR） method， and was cloned into the pc3.1 （+） eukaryotic expression vector. The construction of GACAT2 over-expression vector was confirmed by bacterial liquid PCR method and gene sequencing. The pc3.1-GACAT2 over-expression plasmid and control plasmid pc3.1 were separately transfected into the human lung cancer A549 and H1299 cells. The A549 and H1299 cells with stable over-expression of GACAT2 were established by G418 screening， and the cells were divided into blank vector group （transfected with pc3.1 empty vector） and pc3.1-GACAT2 group （transfected with pc3.1-GACAT2 recombinant plasmid）. RT-qPCR method was used to detect the expression levels of GACAT2 mRNA in the A549 and H1299 cells in two groups；CCK-8 assay was used to detect the proliferation activities of the cells in two groups； cell clone formation assay was used to detect the clone formation numbers of the cells in two groups； stem cell sphere formation experiment was used to detect the sphere formation numbers of the A549 and H1299 cells in two groups. Results The eukaryotic over-expression vector of GACAT2 was successfully constructed. Compared with blank vector group， the expression levels of GACAT2 mRNA in the A549 and H1299 cells in pc3.1-GACAT2 group were increased （P<0.05 or P<0.01）， the cell proliferation activities were decreased （P<0.05 or P<0.01），the clone formation numbers were decreased （P<0.05 or P<0.01）， and the sphere formation numbers were decreased （P<0.05 or P<0.01）. Conclusion Over-expression of human lncRNA GACAT2 gene can inhibit the proliferation and stemness of the lung cancer A549 and H1299 cells.

Objective: To analyze the treatment of a patient with recurrent and refractory small cell lung cancer(SCLC) intolerable further chemotherapy who gained significant efficacy by anlotinib combined with thoracic radiotherapy and to clarify the efficacy and safety of anlotinib combined with radiotherapy in the SCLC patients, and to provide the reference for the selection of treatment program of these patients. Methods: The clinical data of one patient with advanced recurrent and refractory SCLC who could not tolerate further chemotherapy were collected, and the related literatures were reviewed to analyze the efficacy and safety of anlotinib combined with thoracic radiotherapy. Results: The patient was a 70-year-old woman who had a long histroy of hypertention and diabetes,and presented with cough and shortness of breath.Based on the chest CT,biopsy and pleural effusion cytology,the patient was definitely diagnosed as extensive stage of SCLC(pleural metastasis) and then underwent a series of treatments. After three lines of chemotherapy regimens, the patient was unable to tolerate the continued chemotherapy due to the repeated reduce of neutrophil and platelet, and the disease progressed rapidly. Dyspnea aggravated,and the patient was unable to lie. Anlotinib combined with thoracic radiotherapy was initiated.Before radiotherapy,the PET-CT results demonstrated the hypermetabolic lesions including primary tumor located in upper lobe of left lung and bilateral hilar and mediastinal lymph nodes,bilateral pleural effusion.The patient was administrated orally with anlotinib 12 mg once per day for 3 d after thoracic radiotherapy of 6 G/3 f, the patient's wheeze was improved obviously and she could lie flat.However,she complained of dizziness with an increased blood pressure of 168/80 mmHg.Considering that it was related to anlotinib,and the patient's dyspnea was obviously relieved at present to ensure the safe implementation of radiotherapy,so anlotinib was suspended.After irradiation with 38 Gy/19 f,the patient's dyspnea disappeared completely;the primary tumor and metastatic lymph nodes shrank significantly. The CT results showed the intrathoracic lesions were shrank 40 d after radiotherapy;then the patient continued to system therapy. Conclusion: For the patients with SCLC unable to tolerate chemotherapy or progressed after multiple lines of chemotherapy, anlotinib combined with thoracic radiotherapy is promising.In the future,large-scaled clinical trials should be initiated to testify the efficacy and safety of anlotinib combined with thoracic radiotherapy.

Objective To discuss the change of sensitivity of the non-small-cell lung cancer （NSCLC） A549 cells to doxorubicin （Dox） after silencing forkhead protein 3 （FOXP3） gene，and to clarify its mechanism involved in Dox resistance. Methods The human NSCLC A549 cells were transfected with FOXP3 small interfering RNA（siRNA ）by lipofectamine method. The cells were divided into blank control group （without transfection）， si-NC group （transfected with control-siRNA）， and si-FOXP3 group （transfected with FOXP3-siRNA）. Western blotting and immunofluorescence methods were used to detect the expression levels of FOXP3 protein in the A549 cells in various groups；the proliferation activities and half-maximal inhibitory concentration （IC₅₀） values of the A549 cells in various groups were detected by CCK-8 method. The A549 cells were treated with 0， 10， and 20 μmol·L^－1 DAPT， and regarded as 0， 10， and 20 μmol·L^－1 DAPT groups， respectively. Additionally， the A549 cells were treated with 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT， and regarded as 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups， respectively. Western blotting method was used to detect the expression levels of Notch1，Hes1， and FOXP3 proteins in the A549 cells in various groups； the IC₅₀ values and expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 0， 10， and 20 μmol·L^－1 DAPT groups were detected by CCK-8 and Western blotting methods；the expression levels of FOXP3， P-glycoprotein（P-gp）， Notch1， and Hes1 proteins in the A549 cells in 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups were detected by Western blotting method. Results Compared with blank control and si-NC groups， the expression level of FOXP3 protein in the A549 cells in si-FOXP3 group was significantly decreased （P<0.01）， the proliferative activity and IC₅₀ value were decreased （P<0.05 or P<0.01），and the expression levels of Notch1，Hes1 and FOXP3 proteins were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 10 and 20 μmol·L^－1 DAPT groups were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox group were significantly increased （P<0.01）. Compared with 1.0 mg·L^－1 Dox group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT group were significantly decreased （P<0.01）. Conclusion Silencing FOXP3 can enhance the sensitivity of the NSCLC cells to Dox， and its mechanism is related to the inhibition of the Notch1/Hes1 signaling pathway.

Objective To compare the toxicities between gentamicin and its metabolite, and study the effects of Ca²⁺and Mg²⁺ on the toxicity gentamicin. Methods ①The spiral organ of corti of 10 mice (20 ears) were taken, and divided into 2 groups randomly (n=10), cultured in 0.1 mol·L^-1 gentamicin and the metabolite of 0.1 mol·L^-1 gentamicin for 48h, then the number of hair cells remain was recorded;②The spiral organ of corti of 10 mice were taken (20 ears), and divided into 2 groups randomly (n=10), 3% CaCl₂ and 0.1 mol·L^-1 GM were mixed in the culture medium and the number of hair cells remain in the two medium mentioned above were recorded; ③The spiral organ of corti of 10 mice were taken (20 ears), divided into 2 groups randomly (n=10), 3% MgSO₄ to 0.1 mol·L^-1 GM were mixed in the culture medium, the number of hair cells remain in the two medium mentioned above were recorded. Results It was not significant difference between 0.1 mol·L^-1 GM and the metabolite of 0.1 mol·L^-1 gentamicin (P＞0.05) ; but when 3% CaCl₂ or MgSO₄ were mixed in the culture medium, the number of the survived hair cells was observably increased (P＜0.01). Conclusion The toxicity of GM to the cochlea hair cells has nothing to do with its metabolite; Ca²⁺and Mg²⁺ may act as antagonist to the toxicity of GM in cochlea hair cells.

Objective To study the CT, MRI performance and invasion paths outside paranasal sinus malignant tumor. Methods The data of CT and MRI of 31 patients with paranasal sinuses malignant tumor were analyzed, tumor′s invasion scope and invasion paths outside according to the CT, MRI signs and symptoms. Results Paranasal sinus malignant tumor root mainly in maxillary sinus 17　cases (54.8%), then ethmoid sinus 10 cases (32.3%), and frontal sinus 4 cases (12.9%). The CT and MRI results showed soft tissue disease focus was found and its boundary was not clear, enhancement CT and MRI scanning showed 8 and 11 cases had low and middle degrees enhancement,respectively. The pathological changes in 22 cases often be outside the nasal cavity and paranasal sinus but infringed upon the pterygopalatine fossa, orbit. Conclusion CT and MRI can display exactly tumor′s primary focus and its invasion outside the paranasal sinus and judge its invasion path.

To study the apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL) in non-small cell lung carcinoma H596 cells,and to explore the role and mechanism of apoptosis.Methods Non-small cell lung carcinoma H596 cells were cultivated,and acid phosphate assay was used to detect the apoptotic rate,and the expressions of caspase-8,Bcl-2 and Fas-associated death domain (FADD) proteins were detected by Western blotting in H596 cells.Results  The apoptotic rates of H596 cells were decreased after the cells were treated with 0.01-0.03 μg•L^-1 TRAIL,and from 0.10 μg•L^-1 TRAIL,the apoptotic rates were increased;the differences in apoptotic rates of H596 cells after treated  with 10.00-100.00 μg•L^-1 TRAIL were significant compared with control group(P<0.05).The Western blotting results showed that the expressions of caspase-8,Bcl-2,FADD proteins were normal.Conclusion High dose of TRAIL can induce apoptosis in H596 cells,and it may be  related to the normal expressions of   TRAIL related proteins.

Objective To investigate the effects of intrathecal injection with U0126 on the behavior of chronic constrictive injury (CCI) rats and the c-fos expression in spinal cord dorsal horn. Methods Thermal and mechanical nociceptive thresholds were measured before CCI and then on odd-numbered days, up to and including 15th day, to assess and compare the anti-nociceptive effects of adminstration with MEK inhibitor U0126. The time course of expression of c-fos were assessed by immunohistochemical analysis. Results Intrathecal adminstration with U0126 could significantly attenuate the activation of c-fos induced by CCI and simultaneously ameliorate mechanical and thermal hyperalgesia induced by CCI. Conclusion This study shows that the activation of extracellular signal-regulate kinase(ERK) contributes to the CCI-induced neuropathic pain.

Abstract:Objective To reconfirm the association between KPNB3 gene polymorphism with schizophrenia in Chinese population. Methods Two single nucleotide polymorphisms (SNPs), rs2588014 and rs626716 at the KPNB3 locus, were detected in 304 Chinese Han family trios consisting of fathers, mothers and affected offsprings with schizophrenia with PCR-based restriction fragment length polymorphism (RFLP) analysis. The Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness-of-fit test. The UNPHASED program was applied to perform the transmission disequilibrium test (TDT), the haplotype analysis and the pair-wise measure of linkage disequilibrium (LD) between these 2 SNPs. Results The genotypic distribution of both rs2588014 and rs626716 was not deviated from the Hardy-Weinberg equilibrium (P>0.05). The TDT showed significantly biased transmission from parents to affected offsprings for rs626716（χ²=9.31，P=0.002 3）.The allelic frequency transmitted from the heterozygote parents didn’〖KG-3〗t deviate for 50% at rs2588014 (χ²=3.44, P=0.064). The haplotype analysis also showed rs2588104(c)-rs626716(c) was transmitted by parents（χ²=9.8,P=0.006 8）. Conclusion There may be genetic association between KPNB3 gene polymorphism and schizophrenia in the Han descent population in the north of China.

Objective To observe the effect of panax quinquefolium fruit sa ponin (PQFS) on hemodynamics and correlative indexes in the coronary artery liga ted dogs. Methods The dogs were divided into five groups randomly (n= 6): control group,gingkgo leaf tablet group (24 mg·kg^-1), 24,12 and 6 mg ·kg^-1 PQFS groups. The myocardial infarction size (MIS )and the changes o f the serum enzymes were determined by using the acute myocardial infarction mo dels with the ligation of left anterior descending coronary artery of dogs. The parameters of hemodynamics and myocardial oxygen metabolism were measured in the anesthetic dogs with thoracotomy. Results Compared with control group, the myocardial blood f low was increased（P<005）,the coronary resistance and myocardial oxygen w ere decreased（P<005）in dogs treated with PQFS of 24, 12 and 6 mg·kg^-1. The MIS and the activities of serum glutamic oxaloacetict ransaminase enzy me　(AST), creatine kinse (CK) and lactic dehydrogenase (LDH-L) were decreased （P<001,P<005）in acute myocardial infarction dogs treated with PQFS of 24,12 and 6 mg·kg^-1. The activities of superoxide dismutase (SOD) wa s increased（P<001,P<005）, but the content of serum malonyl dialdeh yde (MDA) was decreased（P<001,P<005）. Conclusion PQFS has protective effect on myocardial ischemi a through decreasing myocardial oxygen consumption and increasing myocardial blo od flow in dogs.

Objective To explore the protective effects of oleanolic acid (OLA) derived from roots and leaves of Aralia Elata (Miq) Seem. on alcoholic liver disease (ALD) in rats and its mechanism. Methods Normal rats were intragastrically infused with ethanol and high fat diet. OLA was used from the beginning. Rats were sacrificed after 3 months interference. Hematoxylineosin (HE) method was used to examine the degree of hepatic injury. The concentrations of malandialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were estimated in liver tissue. Blood samples were collected to measure the concentrations of aminotransferase, choline lipase (ChE), triglyceride (TG), total protein (TP), albumin (ALB), bilirubin (BIL), glutathione peroxidase (GSH-Px) and MDA. Results OLA decreased the levels of ALT, AST, TG, ChE, MDA in serum and the level of MDA in liver tissue in model group. In the same time it increased the level of GSH-Px in serum and the levels of SOD and GSH in liver tissue. Hepatic injury was reversed with OLA. Conclusion OLA can protect liver from alcohol injury in rats via eliminating free radicals and decreasing enzymes and blood fat in liver. Injury of free radicals and lipid oxidation are the main pathogenesis of alcoholic liver disease.

Abstract:Objective  To explore the effect of Juglone on proliferation of liver cancer HepG2 cells and to clarify its possible mechanism. Methods The human hepatoma HepG2 cells were divided into different doses of Juglone（40，80，120，160 and 200 μmol/L) groups and control group.The proliferation inhibition of Juglone on HepG2 cells was measured by MTT assay,and IC50 was calculated based on the effective concentration.Optical microscope was used to observe the morphological changes of HepG2  cells．Meanwhile the expression of Caspase-3 protein was analyzed by immunocytochemistry.The cell cycle of HepG2 cells was detected by flow cytometry．Results The IC50 of Juglone was 139.888 1 μmol/L.Compared with control group, the inhibitory rates of proliferation of HepG2 cells in different doses of Juglone groups  were increased significantly(P<0.05 or P<0.01) in a  dose-dependent manner.The morphological observation showed that the cells in different does of Juglone groups were smaller and the  connection disappeared.In 120 μmol/L Juglone group the cell cycle of HepG2 cells  was changed and the cells were  markedly blocked in the G2/M phase.The immunocytochemistry staining results indicated that the Caspase-3 protein  expression in 120 μmol/L Juglone group was higher than that in control group.Conclusion  Juglone could induce the apoptosis of HepG2 cells,and the mechanism may be related to Caspase pathway.

Objective To investigate the possibility of constructing artificial epiphyseal plate using combined scaffold. Methods The jelly-like cartilaginous tissue was constructed by mixing epiphyseal plate chondrocyte, 0.3% collagen Ⅰ, 1 000 U·mL^-1 thrombin and 20 g·L^-1 cryodesiccant human fibrinogen. Paraffin sect ion, HE stain, toluidine blue stain and immunohistochemistry were performed regularly. Results Chondrocyte culture used collagen-fibrous protein combined scaffold was baby pink jelly-like, and the cells distributed uniformly. After 3 days, suspended matter took on milkiness, transmittancy depressed. It contracted to 7 mm while hardness increasing. By the method of HE staining, the chondrocyte was round with hyperchromatism of the nuclear, and cartilage matrix was stained powder blue within cartilage cavities in which homologue cell presen ted. The number of cells decreased gradually from surface layer in which the cel l density was biggest to deep zone. The cells were arranged by strata sequence, but didn′t lined as those of epiphyseal plate. There were brown yellow substanc e in cell periphery showed by immunohistochemistry result, which meaned collagen Ⅱ was positive. Conclusion The effect of constructing artificial epiphyseal plate using the combined scaffold is satisfied.

Objective To investigate the expressions of CA125 and CA19-9 in endometriosis and the relationship with the serum values of CA125 and CA19-9. Methods Serum CA125 and CA19-9 levels were measured by using the magnetic separated enzyme linked immunosorbent assay respectively in 20 patients with sendometriosis. Forty paraffin samples of endometriosis（20 ectopic endometria and 20 endometria）were examined by immunohistochemical staining method to detect the expressions of CA125 and CA19-9. Results CA125 and CA19-9 were mainly expressed in the cytoplasm of glandular epithelial cells. The positive expression rates of CA125 in ectopic endometrium and endometrium were 90.0% and 25.0%, respectively；The expression rates of CA19-9 in ectopic endometrium and endometrium were 85.0% and 10.0%, respectively. The mean serum values of CA19-9 and CA125（U•mL^-1）in 6 patients with stage Ⅰ and Ⅱ endometriosis were (29.64±10.24) and (25.85±9.45),respectively; the positive rates were 16.7% and 33.3%,respectively. The mean serum value of CA19-9 and CA125 （U•mL^-1） in 14 patients with stage Ⅲ and Ⅳ endometriosis were (46.54±14.29 ) and (58.02±20.11), respectively；the positive rates were 42.8% and 71.4%, respectively. The expression of CA19-9 in ectopic endometrium was correlated to the serum value of CA19-9 （r_s=0.36,P<0.05）, but not CA125 (r_s=0.12,P＞0.05). Conclusion The CA125 and CA19-9 proteins express highly in ectopic endometrium, and express lowerly in endometrium. The main resource of serum CA19-9 in patients with endometriosis is ectopic endometrium.

Objective To investigate the value of the expressions of estrogen receptor(ER) and progesterone receptor (PR) in the cast-off cells from the fluid of latex ductal lavage in the diagnosis and treatment of breast diseases. Methods A total of 58 female out-patients were examined by fiberoptic ductoscopy (FDS). All patients were divided into 6 groups by the complaints, physical examination, ultrasound and FDS:normal group (10 persons without symptoms and abnormalities in any examination),galactostasis group (n=19), latex fill up group (n=11), intraductal papilloma group (n=2), mammary ductal ectasia group (n=7) and contralateral examination after mastectomy for breast cancer group(n=9). A total of 168 ducts were successfully examined by FDS, and the expressions of ER and PR in the cast-off cells were detected among all of the samples. Results The positive rates of ER and PR expression in the cast-off cells from the fluid of latex ductal lavage in benign breast disease groups were much higher than those in normal group (P<0.05). The positive rates of ER and PR expression in the cast-off cells from the fluid of abnormal ducts detected by FDS were also much higher than those in normal ducts (P<0.05) in the same case of breast disease. The expression rates of ER and PR in the cast-off cells in normal group showed no significant difference as compared with normal ducts detected by FDS in the abhormal breast. Conclusion The local high expressions of ER and PR are associated with the high risk of breast diseases. The detection of ER and PR expressions plays an important role in guiding the treatment and judging the risk factors of breast diseases.

Objective  To observe the short-term efficacy of GP regimen in adjuvant chemotherapy in patients with non-small-cell lung cancer，and to study t
he factors influencing efficacy and analyze the adverse reactions.
Methods  Clinical data of 68 NSCLC patients received postoperative adjuvant chemotherapy of GP regimen was  retrospectively analyzed.Kaplan-Meier method was used to draw the disease-free survival curve.Log-rank time series analysis was used between different subgroups;C
OX proportional hazards model was used for the univariate and multivariate analyses.
Results  Of all the 68 cases，the median recurrence-free survival time was 33.7 months，56 cases were followed up more than one year，and one year disease-free survival rate was 83.9%;25 cases were followed up more than two years，and
two-year disease-free survival was 72.0%.Univariate analysis suggested that clinical stage(P<0.05)，pathological type(P<0.05),degree of differentiation(P<0.05) significantly influenced the short-term efficacy. Multivariate analysis suggested that clinical stage(P<0.05),degree of differentiation(P<0.05) were thei
ndependent factors of the short-term efficacy.The major adverse reactions were stage 3 to 4 myelo-suppression including neutropenia（33.9%），granulocytopenia（20.6%），anemia（4.4%），and thrombocytopenia（4.4%），and  stage 3 to 4 nausea and vomiting(10.3%).
Conclusion The GP regimen is feasible,well-tolerated with a high disease control rate in one or two years in the adjuvant chemotherapy;clinical stage
and degree of differentiation are the independent factors of the short-term efficacy.

Objective To study the expressions of p16, Bcl-2, Bax, VEGF and significance of microvessel density(MVD) in human pancreatic cancer. Methods Immunohistochemistry method and morphological semi-quantitative analysis were used to detect the quantity of MVD and the expressions of p16,Bcl-2,Bax, and VEGF in 33 cases of pancreatic cancer and 6 normal pancreas tissue specimen. Results The expression of P16 protein: it was higher in moderately or well differentiated pancreatic adenocarcinoma than poorly one (P<0.05), it was higher in patient with long survial time than those with short survial time (P<0.05);the expression of Bcl-2: it was higher in moderately or well differentiated pancreatic adenocarcinoma than poorly one; the expression of Bax:it was higher in normal pancreatic tissue than pancreatic cancer, as well as moderately or well differentiated than poorly one and survival time≥12 months than <12 months; the expression of VEGF:it was higher in patients with lymph node metastasis than without lymph node metastasis; MVD count: it was lower in normal pancreatic tissue than pancreatic cancer as well as with lymph node metastasis than without lymph node metastasis; the expression of MVD had positive correlation with VEGF expression （r=0.79，P<0.05）；p16 gene expression had negative correlation with VEGF expression and MVD （r=-0.69,P<0.05;r=-0.87,P<0.05），but Bcl-2expression had no correlation with other gene expressions. Bax expression had positive correlation with p16 gene expresion （r=0.54,P<0.05），and had negative correlation with VEGF expression and MVD（r=-0.66,P<0.05）. Conclusion The determination of p16, Bcl-2, Bax, VEGF expressions and MVD in human pancreatic cancer may be used to evaluate the prognosis and judge the differentiation of the adenocarcinoma.

Objective To observe cell fusion from without(FFWO) induced by human herpesvirus 6(HHV-6). Methods Cells were overlaid with 10⁵　HHV-6-infected CBMCs (nearly 80% positive for HHV-6 late antigen by IFA) or infected with cell-free HHV-6 at 10^3-4TCID₅₀ for 60 minutes and the cells were stained with HE 2-3 days after infection.To determine whether HHV-6A induced FFWO or not, Vero cells (5×10⁵ per well) were overlaid with 10⁵ HHV-6-infected CBMCs (nearly 90% positive for HHV-6 late antigen by IFA) or infected with cell-free HHV-6 at 10^3-4 TCID₅₀ for 60 minutes, and cultured with medium supplemented with PFA (300 mg•L^-1), fixed 24 hours after infection;Vero or MT4 cells were infected with cell-free virus (HHV-6A or HHV-6B) for 1 hour, and cycloheximide was added to a final concentration of 100 mg•L^-1. After incubation at 37℃ for 2 hours, the cells were fixed and stained with HE. Results The syncytia formation induced by HHV-6A was observed 2 hours after infection; moreover, it was found in the presence of cycloheximide, indicating that HHV-6A induced FFWO in the target cells. Furthermore, the fusion event was dependent on the expression of the HHV-6 entry receptor, CD46, on the target cells membrane. Conclusion HHV-6A can induce FFWO in a variety of human cells through its entry receptor, CD46.

Objective  To establish an indirect sandwich enzyme-linked immunesorbent assay (ELISA) kit for detecting serum level of mucin 1(MUC1) protein and to
explore its application in diagnosis of lung cancer,and to reveal the value of MUC1 protein  detection in clinic.Methods With rabbit anti-human MUC1 monoclonal antibodies and rat anti-MUC1 polyclonal antibodies as coating antibodies and testing antibodies,respectively,a new ELISA method was developed in this study,and the standard curve was gotten.Then,the serum MUC1 levels in the peripheral blood of 48 cases with lung cancer,7 cases with lung benign diseases,and 20 healthy subjects were detected with indirect ELISA.The optimal cut-off value,the specificity,the sensitivity and the Youden index were determined with ROC curve analysis.At the same time,the serum levels of MUC1 detected by CA15-3 commercial kits were analyzed.
Results Based on indirect sandwich ELISA,the cut-off value for serum MUC1 was 1.98  μg?L-1,the sensitivity was 62.5%,the specific
ity was 100% and the Youden index was 0.6250.While based on the CA15-3 kit,the sensitivity was 18.75%,the specificity was 100% and the Youden index was 0.187 5.Conclusion The indirect ELISA kit is superior to the CA15-3 kit in detecting MUC1 in lung cancer .

Abstract:Objective To identify the relationship between the CACNA1H gene and childhood absence epilepsy(CAE).Methods Exons 6 to 12 of CACNA1H gene were sequenced in 100 CAE trios of Han population in China.Variants in these regions were then analyzed.Results Fourteen variants were identified only in 32 CAE patients that were not present in 191 normal gender-matched controls.Twelve of them had not been previously reported.These 14 variants included 2 nonsynonymous variants,3 synonymous variants,9 intronic variants,including a single nucleotide polymorphism (SNP) 52037C>T in intron11 found in 17 CAE patients.The result of transmission disequilibrium test (TDT) indicated that 52037C>T in intron11 was in significant transmission disequilibrium in CAE trios（χ²＝9.783,P=0.001 76）.Conclusion CACNA1H is a susceptibility gene for CAE in the Chinese Han population.

Objective To explore the relations of recurrent spontaneous abortion (RSA) with anticardiolipin antibody (ACA) and antinuclear antibody (ANA). Methods The levels of ACA and ANA in serum of 98 RSA patients and 40 normal women were detected by ELISA. Results The positive rate of ACA in RSA patients (32.7%) was obviously higher than normal wowen (5.0%) (P<0.01);there was no difference of the positive rate of ACA in statistics between early and late RSA, as well as between 2 times and more than 2 times abortions of RSA patients(P>0.05); the positive rates of IgG and IgM were higher among IgM,IgG and IgA of ACA, only 4 RSA patients were positive in all, the positive rate of middle and high level of ACA was 37.5%(11/32), the rest were low levels. The positive rate of ANA in RSA patients (17.3%) was obviously higher than that in normal women (2.5%) (P<0.05). Conclusion ACA and ANA are related with RSA, and may be regarded as a immunological assistant diagnostic index.

To study the healing effect of recombinant human acidic fibroblast growth factor (rhaFGF) expressed in E.coli on dermal chronic ulcers in diabetic rats. Methods　Ten male Wistar rats were used to set up diabetic dermal chronic ulcers models. The wounds were sprinkled with rhaFGF and physiological saline, respectively. The wound area, wound cavity volume and healing time were recorded, granulation tissue growth and epithelization in wound were observed, and wound healing status was evaluated. Results　Compared with control group, the wound area and wound volume at different day in rhaFGF group were significantly diminuted(P＜0.05）, the healing rate was predominantly eleva Ted　(P＜0.05）, the healing time was significantly shortened(P＜0.05）. RhaFGF resulted in a remarkable enhancement of granulation tissue growth and epithelization in wound. Conclusion　The topical application of rhaFGF can significantly accelerate the healing of dermal chronic ulcers in diabetic rats.

Objective To discuss the therapeutic effect of Saposhnikoviae Radix on rheumatoid arthritis and its effect on the nuclear factor-κB （NF-κB） signaling pathway，and to provide the basis for the treatment of rheumatoid arthritis. Methods Fifty rats were randomly divided into control group， model group， dexamethasone group， low dose of Saposhnikoviae Radix group， and high dose of Saposhnikoviae Radix group，and there were 10 rats in each group； except for control group， the rats in the other groups were given intradermal injection of type Ⅱ collagen at the root of the tail and left posterior foot to prepare the rheumatoid arthritis models； after the models were prepared successfully， the rats in dexamethasone group were given dexamethasone （2 mg·kg^－1），the rats in low and high doses of Saposhnikoviae Radix groups were given Saposhnikoviae Radix wild product （0.45 and 0.90 g·kg^－1）， and equal volume of normal saline was given to the rats in control group and model group.The arthritis index（AI） and foot swelling degrees of the rats in various groups were recorded， the organ coefficients of the rats in various groups were calculated； enzyme-linked immunosorben assay （ELISA） method was used to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in serum of the rats in various groups， and the expression levels of NF-κB，NF-κB inhibitor α （IkB-α）， IκB-α kinase β （IKK-β）， cyclooxidase-2 （COX-2），and TNF-α proteins in synovial tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group， the AI score and swelling degree of foot of the rats in model group were significantly decreased （P<0.01）， while the thymus index was significantly increased （P<0.01）； compared with model group， the AI score and swelling degree of foot of the rats in dexamethasone group and low and high doses of Saposhnikoviae Radix groups were significantly decreased （P<0.05 or P<0.01）， and the thymus indexes were significantly decreased（P<0.01）.The ELISA results showed that compared with control group，the levels of TNF-α， IL-1β，and IL-6 in serum of the rats in model group were increased（P<0.01）； compared with model group，the levels of TNF-α， IL-1β，and IL-6 in serum of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were significantly decreased （P<0.01）.The HE staing results showed that the synovial tissue of the rats in control group was normal，and the synovial tissue of the rats in model group showed hyperplasia accompanied with extensive infiltration of inflammatory cells； compared with model group， the degrees of proliferation of synovial tissue and infiltration of inflammatory cells of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were decreased.The Western blotting results showed that compared with control group， the expression levels of NF-κB，IKK-β，COX-2， and TNF-α proteins in synovial tissue of the rats in model group were increased （P<0.01），the expression level of IκB-α protein was decreased （P<0.01）；compared with model group， the expression levels of NF-κB，IKK-β，COX-2， and TNF-α proteins in synovial tissue of the rats in dexamethasone group， low and high doses of Saposhnikoviae Radix groups were decreased （P<0.01），the expression level of IκB-α protein was increased （P<0.01）. Conclusion Saposhnikoviae Radix wild product can improve the joint injury by reducing the levels of proinflammatory cytokines and inhibiting the excessive activation of NF-κB signaling pathway in synovial tissue， thus playing a therapeutic role in the rheumatoid arthritis.

Objective To explore the pathogenesis of type 1 diabetes mellitus (T1DM) induced by different doses of streptozotocin (STZ) and its relations with autoimmune response. Methods Forty KM mice were divided into control group (group Ⅰ) and 20,40,80 mg·kg^-1 STZ treated groups(group Ⅱ,Ⅲ,Ⅳ). Animal models of diabetes mellitus were induced by different doses of STZ through intra peritoneal injection. Blood and urine glucose (Glu) levels were detected with glucose detecting kit and urine analyzing papers respectively. Pancreatic histological changes of islets were observed under light microscope. The level of insulin autoantibodies (IAA) in serum was detected by indirect ELISA. The functions of Tlymphocytes in mouse spleen and thymus were detected by Lymphocytes Proliferation Assay (MTT method). Results The blood glucose in group Ⅰ changed nothing. The blood Glu levels in group Ⅱ and Ⅲ increased as times went by. Four weeks after injection of STZ, the urine Glu in group Ⅰ were all negative, in group Ⅱ 5 negative and others >+, in group Ⅲ 2 negative and others >++, in group Ⅳ all >+++. The changes and degrees of pancreatic islet β cells injured by difference doses of STZ were different. The IAA levels in groupⅡand Ⅲ were higher than that in control group　(P<0.05), but there was no difference between group Ⅳ and control group. Spleenocytes (mature T lymphocytes) were highly proliferated by ConA and the thymocytes (immature T lymphocytes) were lowly proliferated by Con A in groupⅡand Ⅲ. However, there was no difference between group Ⅳ and control group. Conclusion Three different doses of STZ (20,40,80 mg·kg^-1) can induce different kinds of DM. High dose 80 mg·kg^-1) STZ injection can induce T2DM in mice, and low dose (20,40 mg·kg^-1) and multiple injection can induce T1DM. The better dose is 40 mg·kg^-1.

Objective To study the expressions of p16 and p15 in human endometrial carcinoma，atypical hyperplasia of endometrium, and normal endometrium and its significances. Methods The expressions of p16 and p15 was examined in 57 specimens of malignant, 8 specimens of borderline,7 specimens of benign uterine endometrial tissues and 7 specimens of normal endometrium by immunohistochemistry S-P methods. Results The positive rates of p16 expression in the malignant and non-malignant tissues were 40.4% and 77.3%, respectively, significant difference was observed (P<0.05). The expression of p16 was associated with pathological types, pathological grades and operation-pathology stages, but no association was found with muscle infiltrations. The positive rates of p15 expression in the malignant and non-malignant tissues were 47.4% and 86.3%, respectively, significant difference was observed (P<0.05). The expression of p15 was associated with pathological grades, but no association was found with pathological types, muscle infiltrations, and operation-pathology stages. There was positive correlation between p16 expression and p15 expression in uterine endometrial carcinoma（r=0.91，P<0.05）. Conclusion Low expressions of p16 and p15 may contribute to the development of uterine endometrial carcinoma.

Objective To observe the curative effects of deep partial thickness derma burn injury using bone marrow mesenchymal stem cells. Methods The bone marrow stem cells were obtained from the male guinea pig and cultured in vitro. Then the mesenchymal stem cells (MSCs) with 2×10⁶ cell•mL^-1 and 2×10⁷ cell•mL^-1 were prepared, and transplanted into the wounds of the female guinea pig models with deep partial thickness derma burn injury. The wound healing area was counted and the morphological changes were observed by using HE and Masson stain. Results The wound healing in therapy groups were significantly better than that in control groups. HE stain showed that dermal and epidermal structures were clear, and cell structures of every epidermal layers were distincted and integrated in therapy groups,but cell structures of every layers in control groups were unclear. Masson stain showed that collagen was regular and new-born epidermal covered the wound in therapy groups, collagen was disorder and thicker in control groups. Bone marrow MSCs could promote the healing of wound and the effects in therapy groups were better than that in the control groups. But there was no significantly difference between MSCs with 2×10⁶ cell•mL^-1 and 2×10⁷ cell•mL^-1 in therapy groups. Conclusion The bone marrow MSCs have significant curative effects on deep partial thickness derma burn injury.

Abstract:Objective
To construct the recombinant vectors that coexpress wild type p53(wt-p53) and siRNA-mdm2 for gene therapy in androgen independent prostate cancer .Methods The subcloning,T-Acloning,and PCR technique were used to construct the recombinant vectors,named pcDNA3.1-p53,siRNA-mdm2,p53/siRNA-mdm2, and siRNA-scramble .The recombinant expression vectors mentioned above were transfected into PC-3 cells,the expression levels of the mdm2 siRNA and p53 after transfection were detected by the semi-quantitative RT-PCR analysis and Western blotting analysis.MTT assay was used to detect the  proliferation inhibition  of PC-3 cells after transfection.Results The recombinant plasmids containing both wt-p53 and siRNA-mdm2 were successfully constructed,and confirmed by DNA sequence analysis.After transfected for 48 h, the expression of GFP was observed.The expression level of wt-p53 was increased,and the expression of mdm2 was decreased detected by RT-PCR and Western blotting.MTT assay showed that the inhibitory rate of  proliferation of PC-3 cells was 40.4% after transfection.Conclusion The recombinant plasmids containing both wt-p53 and siRNA-mdm2 are successfully constructed,they can remarkablely inhibit the proliferation of PC-3 cells.

Objective To screen the differentially expressed genes in type 2 diabetes mellitus model rats by transcriptomics sequencing， and to preliminarily explore the relationship between the differentially expressed genes and the syndrome manifestations of deficiency of both Qi and Yin. Methods Eighteen male SD rats were randomly selected as control group，and the other rats were fed with high-fat and high-sugar diet and injected with streptozotocin （STZ） in the abdomen to establish the type 2 diabetes models. On the 6th week， 31 successfully modeling rats were divided into model group （n=14） and drug evidence group （n=17） according to the principle of blood glucose and body mass balance. The rats in model group were fed with high sugar and high fat diet sequentially.The rats in drug evidence group were given Xiaokeling 2.2 g·kg^－1·d^－1 by gavage；the experiment last for 21 weeks.The body masses， food intakes， water intakes， mental state scores， sport scores， foot temperatures， grasping force， pulse amplitudes， respiratory rates， and tongue image integral of the rats were detected and the physiological signs of the rats during model preparation were evaluated.The fasting blood glucose（FBG） levels of the rats in various groups were detected by blood glucose meter. After the last administration， the rats were anesthetized， the blood samples were collected and the serum samples were separated.The levels of triglyceride （TG） and total cholesterol （TC） of the rats were detected by oxidase method.The levels of high density lipoprotein cholesterol （HDL-c） and low density lipoprotein cholesterol（LDL-c）of the rats were detected by direct method，and the levels of CD4， CD8，cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） of the rats were detected by enzyme-linked immunosorbent assay （ELISA） method.The liver tissue of the rats was obtained，and the differentially expressed genes between model group and control group and between model group and drug evidence group were screened by transcriptomic sequencing. Results Compared with blank group， the physiological signs of the rats in model group were basically consistent with the syndrome manifestations of Qi and Yin deficiency； compared with model group， the some physiological signs of Qi and Yin deficiency of the rats in drug evidence group were significantly improved. Compared with blank group， the FBG，serum TC， TG and LDL-c levels of the rats in model group were significantly increased （P<0.01）， and the serum HDL-c level had no significant difference（P>0.05）；compared with model group， there were no significant differences in the above indexes of the rats in drug evidence group （P>0.05）.Compared with blank group， the level of serum CD4 and the CD4/CD8 ratio of the rats in model group were significantly decreased （P<0.01）， while the level of serum CD8 had no significant difference（P>0.05）；the cAMP level in serum and the ratio of cAMP/cGMP were obviously increased （P<0.01）， while the cGMP level in serum was significantly decreased （P<0.01）；compared with model group， the level of serum CD4 and the CD4/CD8 ^{ratio of the rats in drug evidence group were remarkably increased （P<0.01）， the cGMP level in serum was increased（P<0.01） and the cAMP/cGMP ratio was significantly decreased（P<0.01）， the cAMP level in serum had no significant difference （P>0.05）. Compared with blank group， the up-regulated expressed genes of the rats in model group mainly included early growth response factor 2 （Egr2），insulin-like growth factor binding protein 1 （Igfbp1） and a disintegrin and metallopeptidase with thrombospondin motifs 4 （Adamts4）； the down-regulated expressed genes mainly included G₀/G₁ switch 2 （G0s2）， midline 1 interaction protein 1 （Mid1ip1）， and basic helix-loop-helix family e40 （Bhlhe40）；compared with model group， the levels of the above differentially expressed genes in drug evidence group showed the opposite trend. Conclusion The syndrome of type 2 diabetes mellitus combined with deficiency of Qi and Yin is associated with up-regulation of Egr2， Igfbp1 and Adamts4 and down-regulation of G0s2， Mid1ip1，and Bhlhe40.}

Objective To investigate the effects of Gross Saponin of Tribulus Terrestris (GSTT) on acute myocardial ischemia in rats. Methods The acute myocardial ischemia models were established by isoproternol subcutaneous injection and pituitary sublingual venous injection respectively. The effects of GSTT on the electrocardiogram T-wave of isoproternol inducing acute myocardial ischemia and the serum contents of LDH and CK of pituitary inducing acute myocardial ischemia were observed. Results GSTT with doses of 10.4,20.8,and 31.2 mg•kg^-1 could significantly improve the decreased heart rates induced by isoproternol (P<0.01, P<0.05);GSTT with dose of 31.2 mg•kg^-1 could remarkably increase the activities of serum LDH and CPK in acute myocardial ischemia rats (P<0.01).Conclusion GSTT can protect myocardium in rats with acute myocardial ischemia.

Objective To detect the association between the polymorphism of tumor necrosis factor promoter gene (TNF-308) and asthma. Methods The technique of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was used to examine the allele frequencies of TNF-308 in 50 asthma patients and 80 normal controls. Results The allele frequency of TNF-308 gene was not significantly different between asthma group and normal control group （χ²=0.047, P>0.05）. Meanwhile, the frequencies of three genotypes （GG,GA and AA）of TNF-308 gene were not significantly different in asthma group (45/50, 5/50 and 0/50) and normal control group (71/80, 9/80 and 0/90)（χ²=0.049,P>0.05）. Conclusion There is not association between the polymorphism of tumor necrosis factor promoter gene (TNF-308) and asthma.

To construct a CTL epitope vaccine against human papillomavirus and to express it in E. coli. Methods The gene sequences encoding CTL epitopes selected by computer-assisted epitope prediction were synthesized by PCR and sub-cloned into pET28a expression vector. The recombinant plasmid pET28a-Tat-HPVepi was transformed to E. coli for expression and the expressed protein was identified by Western blotting. Results The DNA sequences of selected CTL epitopes were obtained by 5 rounds of PCR, the constructed recombinant plasmid pET28a-Tat-HPVepi was effectively expressed in E. coli. The expressed products，identified by Western blotting，showed specific bands. Conclusion The CTL epitope vaccine against human papillomavirus is successfully constructed and express in E. coli.

Objective To investigate the expressions of C-erbB-2, Nm23 and MDR1 in primary breast cancer and their relations with metastasis and prognosis. Methods Immunohistochemical method (S-P method) was used to detect C-erbB-2, Nm23, MDR1 expressions in 49 patients with primary breast cancer. The correlations between the three genes and their relations with metastasis and prognosis were analysed. Results The metastasis rate was higher in patients with high-expressions of C-erbB-2, MDR1 and low-expression of Nm23（P<0.05）. The expressions of C-erbB-2 and MDR1 had correlation (r_s=1.00, P<0.05). The expression of C-erbB-2 and Nm23 had no correlation（r_s=0.80，P>0.05）. The expression of MDR1 and Nm23 had no correlation（r_s=0.80，P>0.05）. The metastasis rate in patients with both C-erbB-2(+) and MDR1(+) was higher than the others. Their prognosis was negative. The metastasis rate in patients with both C-erbB-2(+) and Nm23(-) was higher than the others. Conclusion There are correlation between C-erbB-2, Nm23, MDR1 expressions and metastasis of breast cancer. Multi-predictors are better guide to patients than single predictor.

Objective To study the relationships between prognosis and expressions of p53,c-erbB-2, p21 and nm23 genes in gastric cancer tissue specimens and evaluate their values in diagnosis and treatment of gastric cancer. Methods The expressions of p53, c-erbB-2, p21 and nm23 were detected by immunohistochemical method in 123 surgically resected specimens. Results The positive expression rates of p53,c-erbB-2,p21 and nm23 in well-differentiation gastric cancer group were 38.0%(27/71)， 59.2%(42/71)， 39.4%(28/711) and 71.8%(51/71)， respectively；the positive rates in poor-differentiation group were 46.2%(24/52),36.5%(19/52), 69.2%(24/52) and 34.6%(18/52)，respectively.Expressions of p53，c-erbB-2，p21 and nm23 were not observed in non-tumor endoscopic biopsy specimens. The expressions of p53，c-erbB-2 ，p21 were positively correlated with grade of tumor differentiation ，the depth of tumor invasion and clinical different stages (r=0.53, r=0.67, r=0.48, P<0.05), and the expression of nm23 was negatively correlated with grade of tumor differentiation，the depth of tumor invasion and clinical different stages　(r=-0.36, P<0.05). Conclusion Determination of p53, c-erbB-2, p21 and nm23 might be useful in diagnosis and treatment of gastric cancer, differentiating benign and malignant tumor, clinical stages and better than single gene in predicting prognosis of gastric cancer. Key words：stomch neoplasms；gene p53； recepto r， c-erbB-2；gene， c-erbB-2，p21， nm23； prognosis

Objective  To investigate the effects of different target concentrations of propofol on mRNA expressions of pro-inflammatory cytokines as interleuk
in-1（IL-1）,interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in patients underwent cardiopulmonary bypass (CPB) surgery,and to explore the ideal concentration of propofol in Chinese people for clinical use. Methods Sixty patients underwent CPB were randomly assigned into three groups (n=20) according to the different target concentrations:group A,2 mg?L-1; group B,3 mg?L-1;group C,4 mg.L^-1.The blood samples were taken at the time of induction(T1),right after CPB(T2),30 min after CPB(T3),60 min after CPB(T4) and 3 h after CPB(T5).The mRNA expressions of cytokines were detected by RT-PCR.Results  The levels of the serum cytokine expressions of the patients in three groups showed no difference before C
PB,while the levels of all cytokines rushed up after CPB and went up at peak between 30-60 min after CPB,which had great differences at T1,T2,T3 and
T4 compared with T0.Compared with group A,the levels of all cytokines in group B and group C  were significantly decreased(P<0.05).However,the expression levels of cytokines  had no difference between group B and group C. Conclusion Propofol can inhibit the expressions of serum pro-inflammatory cytokines,including IL-1,IL-6 and TNF-α,in patients underwent CPB surgery;therefore inhibit inflammatory reaction.The recom
mended target concentration of propofol would be 3 mg.L^-1 in CPB surgery. 

Objective To investigate the antimicrobial effect of silver carboxymethyl chitosan (CMCT-Ag⁺) on porphyromonas gingivalis (Pg), a critical bacteria in periodontitis and provide experimental basis for developing higher efficiency and lower side-effect drug to cure periodontitis. Methods CMCT was alkalified, modified with chloracetic acid and exchanged ion to obtain CMCT-Ag⁺ complexation. The agar diffusion and plate serial dilution test was used to detect the effect of CMCT-Ag⁺ against Pg by observing cingula diameter and bacteria plaque. Results The pink CMCT-Ag⁺ was obtained, yield ratio was 89.4 %, the degree of substitution of CMCT was 0.98 and the content of Ag⁺ was 10.21% in the obtained CMCT-Ag⁺. In the agar diffusion test, when concentration of CMCT-Ag⁺ exceed 0.3125 g•L^-1, the bacteriostasis ring diameter was increased, and at concen tration of 40 g•L^-1, the bacteriostasis ring diameter inhibited by CMCT-Ag⁺ was 9.5 mm，that was approximation to the result induced by 0.005 g•L^-1 tinidazole. But when concentration of CMCT-Ag⁺ exceed 40 g•L^-1, the bacteriostasis ring diameter was decreased. These results showed that the inhibitory concentration of CMCT-Ag⁺ was more than 0.3125 g•L^-1. In the plate serial dilution test, when concentration of CMCT-Ag⁺ exceed 0.3125 g•L^-1, the growth of Pg was obviously inhibited on medium, and when concentration of CMCT-Ag⁺ exceed 1.25 g•L^-1, the growth of Pg couldn’[KG-*3]t be found. These results indicated that the minimal inhibitory concentration (MIC) of CMCT-Ag⁺ was 1.25 g•L^-1. Conclusion CMCT-Ag⁺ could inhibit the growth of Pg, hence it can be used for periodontal treatment.

Objective To investigate the curative effect of osimertinib combined with savolitinib targeted therapy in the patients with non-small cell lung cancer（NSCLC） and central nervous system（CNS） metastasis with acquired resistance to epidermal growth factor receptor（EGFR）-tyrosine kinase inhibitor（TKI）， and to provide the reference for the targeted therapy for the disease. Methods The clinical data of one NSCLC complicated with CNS metastasis patient with non-frameshift deletion mutation in exon 19 of the EGFR gene complicated with MET amplification were collected， and the clinical characteristics， treatment methods， drug resistance mechanisms of targeted therapy， and treatment methods after drug resistance were analyzed combined with the literature review. Results The patient， female，47 years old， was diagnosed as lung adenocarcinoma in our hospital 3 years ago. The genetic test results showed that there was a non-frameshift deletion mutation in exon 19 of the EGFR gene， and the patient was given icotinib targeted therapy. After 18 months of treatment， the genetic test results showed the patient had the Exon20-T790M mutation， and the patient received oral osimertinib targeted therapy. Twelve months later， the disease progressed and CNS metastasis was found by the re-examination， and the genetic test showed the patient had the mesenchymal-epidermal transforming factor（MET） amplification，the patient received oral osimertinib combined with anlotinib targeted therapy. More than 2 months later， the re-examination results found that the disease had further progress， and the treatment plan was adjusted into osimertinib combined with savolitinib targeted therapy. After 1 month and 4 months， the re-examination results of chest CT and head MRI showed that the lung lesions and brain lesions were significantly smaller than before. Conclusion For the NSCLC brain metastases patients with EGFR-TKI resistance complicated with MET amplification after long-term targeted therapy， the combination of osimertinib and savolitinib targeted therapy can significantly control the progress of the lung and CNS disease in the short term.

Objective To obtain the E2 genes from Neimeng strain of classical swine fever virus and construct fusion genes. Methods E2 DNA fragment was acquired from spleen of infected pig and amplified through RT-PCR technique. After being sequenced, the E2 gene was ligated to Chaperone 10 to form the fusion gene Chap 10-E2, the fusion gene was ligated to expressing vector. Results There were differences in four bases between E2 gene from Neimeng strain of swine fever virus and E2 gene from HCLV strain. The fusion gene Chap 10-E2 was successfully constructed. Conclusion The difference of E2 gene of swine fever virus from different strains is not significant.

Objective To study the changes of serum leptin in patients with simple obesity and patients with type 2 diabetes mellitus complicated with obesity in order to explore the relationship of leptin and insulin resistance and the role of leptin in the occurrence of type 2 diabetes mellitus. Methods 60 cases of simple obesity, 60 cases of type 2 diabetes mellitus and 30 cases of normal control were included according to the diagnostic criteria of obesity and type 2 diabetes mellitus. The levels of fasting serum leptin, fasting serum insulin, fasting glucose, fasting blood lipid were measured in all cases.The body mass index (BMI) and insulin action index were calculated. Results The level of BMI, serum leptin, serum insulin, blood lipid were significantly higher in patients with simple obesity and with type 2 diabetes mellitus complicated with obesity than in normal control cases, whilel (IAI) was significantly lower. The levels of free serum leptin, serum insulin, free glucose, and blood lipid were significantly higher in patients with type 2 diabetes mellitus complicated with obesity than in patients with simple obesity, while IAI was significantly lower. The level of serum leptin was positively correlated with BMI(r=0.48,P<0.55) and fasting serum leptin(r=0.55,P<0.05) and negatively correlated with IAI(r=-0.47,P<0.05)in patients with type 2 diabetes complicated with obesity. Conclusion The overexpression of serum leptin may play an important role in the occurrence of in insulin resistance and type 2 diabetes mellitus in obesity patients.

Objective To study the inhibitory effect of 20(S)-ginsenoside Rg3 on tumor growth and its mechanism. Methods MTT assay and the model of B16 melanoma were used to observe the effect of 20(S)-ginsenoside Rg3 on B16 melanoma growth and its mechanism was analysed by assay of tumor-induced angiogenesis and flow cytometry. Results In vivo, the average weight of B16 melanoma in each Rg3 group was lower than that in control group (P<0.01) and the number of vesssels oriented toward the tumor mass was less than that in untreated control (P<0.01). In vitro, the results of MTT assay displayed that ginsenoside Rg3 inhibited the growth of B16 melanoma cells and the IC₅₀ value was （7.76±0.46）mg·L^-1. Rg3 blocked B16 melanoma cells in G₀/G₁ and S stages. Conclusion 20(S)-ginsenoside Rg3 can obviously inhibit the tumor growth and the effect may be related to its inhibiting angiogenesis and blocking cell mitosis.

Objective To discuss the protective effect of Modified Xiao-Xian-Xiong Decoction （MXD） on liver injury in the rats with type 2 diabetes mellitus（T2DM）， and to clarify its possible mechanism. Methods Fifty rats were fed with high sugar and high fat combined with intraperitoneal injection of streptozotocin （STZ） to establish the T2DM rat models.A total of 40 rats successful modeling were randomly divided into model group（given no drugs），metformin group（given 200 mg·kg^－1 metformin）， low dose of MXD group （given 630 mg·kg^－1 MXD），and high dose of MXD group（given 1 260 mg·kg^－1 MXD）， and there were 10 rats in each group； another 10 rats were selected as control group（given standard feed）. The body masses of the rats in various groups were detected；the fasting blood glucose （FBG） levels of the rats in various groups were detected； the activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum of the rats in various groups were detected by fully automated biochemical analyzer；the pathomorphology of liver tissue of the rats in various groups were detected by HE staining； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin 1β （IL-1β）， interleukin 6 （IL-6）， tumor necrosis factor α （TNF-α） in serum； the levels of superoxide dismutase （SOD）， reductive glutathione （GSH）， and malondialdehyde （MDA） in liver tissue of the rats in various groups were measured by spectrophotometer；the expression levels of nuclear factor E2 related factor 2 （Nrf2） and heme oxygenase （HO-1） proteins in liver tissue of the rats in various groups were detected by Western blotting method. Results The rats in control group had moderate feeding and water intake，and the above indexes of the rats had no sudden increasing and decreasing；the rats in model group had typical symptoms of diabetes；compared with model group，the diabetes symptoms of the rats in meftormin group，low dose of MXD group，and high dose of MXD group had been improved to various degrees.Compared with control group， the body masses of the rats in model group were significantly decreased at different time points（P<0.01）， while the FBG levels were increased （P<0.01）； compared wtih model group，the levels of FBG of the rats in meftormin group，low dose of MXD group，and high dose of MXD group were decreased in the 2nd，3rd and 4th weeks after administration，and the levels of FBG of the rats in low dose of MXD group were decreased in the 3rd and 4th weeks after administration（P<0.05 or P<0.01）.Compared with control group，the activities of AST and ALT in serum of the rats in model group were increased（P<0.01）；compared with model group，the activities of AST and ALT in serum of the rats in metformin group，low dose of MXD group，and high dose of MXD group were decreased（P<0.01）.The HE staining results showed that the morphology of liver cells of the rats in control group was normal，and there was no inflammatory cell infiltration；there was serious inflammatory cell infiltration in model group；there was a bit of inflammatory cell infiltration in meftormin group；there were almost no inflammatory cell infiltrations in low and high doses of MXD groups.The ELISA method results showed that the levels of serum IL-1β， IL-6， and TNF- α in serum of the rats in model group were significantly increased （P<0.01）.Compared with model group， the levels of serum IL-1β， IL-6， and TNF-α of the rats in metformin group， low dose of MXD and high dose of MXD groups were decreased（P<0.01）， while the activity of SOD， the level of GSH， and the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in model group were significantly decreased （P<0.01）.The spectrophotometer results showed that compared with control group， the activity of SOD， the level of GSH in liver tissue of the rats in model group were decreased（P<0.01）， and the level of MDA was significantly increased （P<0.01）；compared with model group，the activities of SOD and the levels of GSH in liver tissue of the rats in metformin group，low dose of MXD group，and high dose of MXD group were decreased（P<0.01），and the levels of MDA in liver tissue of the rats in low and high doses of MXD groups were decreased（P<0.05 or P<0.01）.The Western blotting results showed that compared with control group，the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in model group were decreased（P<0.01）；compared with model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the rats in metformin group，low dose of MXD group，and high dose of MXD group were increased（P<0.05 or P<0.01）. Conclusion MXD has protective effect on liver injury of the rats with type 2 diabetes， and its mechanism may be related to anti-inflammation and activation of Nrf2/HO-1 signaling pathway，thereby inhibition of oxidative stress.

To investigate the therapeutic effects of glucagon-like peptide-1 （GLP-1） analog on rats with experimental diabetes mellitus.Methods Eighty-four Wistar rats were divided randomly into normal control group，high fat forage group，low-dose GLP-1 analog group，middle-dose GLP-1 analog group，high-dose GLP-1 analog group,diabetes mellitus model group and positive control group.Diabetes mellitus animal models were induced with high fat forage and injection of 40 mg•kg^-1 STZ.After 7 and 14 d of GLP-1 analog therapy，the level of fasting plasma glucose(FPG) was detected with enzyme-chemical method.After 14 d of GLP-1 analog therapy，the levels of serum total cholesterols (TC)，triglyceride (TG)，low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were detected with enzyme-chemical method. The levels of serum insulin and C-peptide were measured with immunoradiometric assay method .Results After 14 d of GLP-1 analog injection，food intake in high-dose GLP-1 analog（100 μg•kg^-1）group was less than that in DM group（P<0.01）; Water intake in middle-dose GLP-1 analog（10 μg•kg^-1） group and high dose （100 μg•kg^-1）group were less than that in DM group（P<0.01）. There were no statistical differences of body weight between all therapeutic groups and DM group(P<0.05).The levels of FPG in middle-dose GLP-1 analog（10 μg•kg^-1 ） group and high dose （100 μg•kg^-1）group were lower than that in DM group（P<0.01）.The level of TG in high-dose GLP-1 analog（100 μg•kg^-1）group was lower than that in DM group（P<0.01）.The levels of LDL in middle-dose GLP-1 analog（10 μg•kg^-1） group and high dose （100 μg•kg^-1）group were lo wer than that in DM group（P<0.01）.The levels of serum insulin and C-peptide in middl e-dose GLP-1 analog（10 μg•kg^-1） group and high dose （100 μg•kg^-1）group were higher than that in DM group（P<0.05）. Conclusion GLP-1 analog can paly an important role in treatment of experimental diabetes mellitus through multiple ways.

Objective To discuss the effect of miR-93-5p on the proliferation， migration， and invasion of the rheumatoid arthritis fibroblasts-like synoviocytes（RA-FLSs），and to elucidate its possible mechanism. Methods The rheumatoid arthritis（RA） patients （RA group，n=37） and joint trauma patients who underwent joint replacement surgery （control group，n=30） were selected as the subjects.The RA-FLSs were isolated from synovial tissue of the RA patients， and identified by immunofluorescence and flow cytometry. The RA-FLSs were divided into blank group， mimics NC group （transfected with miR-93-5p mimics NC）， mimics group （transfected with miR-93-5p mimics）， OE-NC group （transfected with ROCK2 over-expression empty plasmid）， OE-Rho related spiral coil protein kinase （ROCK）2 group （transfected with ROCK2 over-expression plasmid）， mimics+OE-NC group （co-transfected with miR-93-5p mimics and ROCK2 over-expression empty plasmids），and mimics+OE-ROCK2 group （co-transfected with miR-93-5p mimics and ROCK2 over-expression plasmids）. Real time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-93-5p and ROCK2 mRNA in synovial tissue of the patients in two groups and cells in various groups；CCK-8 method was used to detect the proliferation activities of the cells in various groups； EdU staining was used to detect the EdU positive rates of the cells in various groups；Transwell champer assay was used to detect the migration and invasion numbers of RA-FLSs in various groups； Western blotting method was used to detect the expression levels of ROCK2， Ki-67， proliferating cell nuclear antigen （PCNA），matrix metalloproteinase 2 （MMP-2）， and matrix metalloproteinase 9 （MMP-9） proteins in the cells in various groups； the targeting relationship between miR-93-5p and ROCK2 was verified by double Luciferase reporter gene experiment. Results The immunofluorescence assay results showed that the expression of Vimentin protein was positive.The flow cytometry detection results showed that the expression of CD55 on surface of the third generation RA-FLSs was positive， while the expressions of CD14 and CD68 were negative， confirming that the isolated cells were the RA-FLSs. The RT-qPCR results showed that compared with control group， the expression level of miR-93-5p in synovial tissue of the patients in RA group was significantly decreased（P<0.01）， while the expression level of ROCK2 mRNA was significantly increased （P<0.01）. Compared with blank group and mimics NC group， the expression level of miR-93-5p of the cells in mimics group was significantly increased （P<0.01）， while the expression levels of ROCK2 mRNA and protein were significantly decreased （P<0.01）. The CCK-8 method and EdU staining results showed that compared with mimics NC group， the proliferation activitiy and EdU positive rate of the cells in mimics group were significantly decreased （P<0.01）， while the proliferation activitiy and EdU positive rate of the cells in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the proliferation activitiy and EdU positive rate of the cells in mimics+OE-ROCK2 group were increased （P<0.01）. The Transwell champer assay results showed that compared with mimics NC group， the migration and invasion numbers of RA-FLSs in mimics group were significantly decreased （P<0.01）， while the migration and invasion numbers of RA-FLSs in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the migration and invasion numbers of RA-FLSs in mimics+OE-ROCK2 group were increased （P<0.01）. The Western blotting method results showed that compared with mimics NC group， the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in mimics group were significantly decreased （P<0.01）， while the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in OE-ROCK2 group were significantly increased （P<0.01）； compared with mimics group， the expression levels of ROCK2， Ki-67， PCNA， MMP-2， and MMP-9 proteins in the cells in mimics+OE-ROCK2 group were significantly increased （P<0.01）.There was a targeted binding site between miR-93-5p and ROCK2-3'-UTR. The double luciferase report experiment results showed that transfection of miR-93-5p mimic could significantly decrease the luciferase activity of the cells in ROCK2 wild type（ROCK2-WT） group（P<0.01）. Conclusion Over-expression of miR-93-5p inhibits the proliferation， migration， and invasion of the RA-FLSs targeted by down-regulation of ROCK2 expression.

Objective To establish a method to isolate and identify human mensenchymal stem cells(HMSC). Methods The HMSC were isolated and cultured in vitro by applying methods of density-grad centrifugal and anchoring screening. The immunocytochemistry, cytochemistry, and electron microscopy were used to identify HMSC cultured in vitro. Results The HMSC have particular phenotype:CD29 and CD44 were positive,CD34 and CD45 were negative.PAS was positive,and SB was negative.HMSC were infantile under electron microscope. Conclusion The HMSC can be isolated with density-grad centrifugal and anchoring screening. The method to identify HMSC with immunocytochemistry, cytochemistry and electron microscopy is effective.

Objective To discuss the expression levels of long non-coding RNA paired box 8 antisense RNA 1 （PAX8-AS1） in the human colorectal cancer （CRC） tissue and its effects on the proliferation， apoptosis and invasion of the CRC cells，and to charify its mechanism. Methods The expression levels of PAX8-AS1 mRNA and miR-22-3p in cancer tissue and cancer cells of the CRC patients were detected by real-time fluorescence quantitative PCR （RT-qPCR） method.The PAX8-AS10 siRNA small molecule sequences and miR-22-3p inhibitors were transfected alone or co-transfected into the CRC cells， and the cells after transfection were divided into si-NC group （transfected with negative sequences）， si-PAX8-AS1 group （transfected with PAX8-AS1 siRNA）， si-PAX8-AS1+inhibitors NC group （co-transfected with PAX8-AS1 siRNA and inhibitors NC）， and si-PAX8-AS1+ inhibitors group （co-transfected with PAX8-AS1 siRNA and miR-22-3p inhibitors）， and the SW480 cells without any transfection were regarded as control group. The expression levels of PAX8-AS1 mRNA and miR-22-3p in the cells in various groups after transfection were measured by RT-qPCR method； the proliferation activities of the cells in various groups were detected by MTT method； the migration and invasion rates of the cells in various groups were measured by Transwell chamber assay； the apoptotic rates of cells in various groups were measured by flow cytometry； the expression levels of B-cell lymphoma-2（Bcl-2）， Bcl-2 associated X protein（Bax）， and cleaved Caspase-3 proteins in the cells in various groups were measured by Western blotting method； the target relationship between PAX8-AS1 and miR-22-3p was verified by dual luciferase reporter gene assay. Results The RT-qPCR results showed that compared with adjacent tissue，the expression level of PAX8-AS1 mRNA in cancer tissue was significantly increased（P<0.01）， and the expression level of miR-22-3p was significantly decreased（P<0.01）； compared with the human normal colonic epithelial cells NCM460，the PAX8-AS1 mRNA expression levels in the SW480， SW620， HT-29 and LoVo cells were all significantly increased （P<0.01）， while the miR-22-3p expression levels were all significantly decreased （P<0.05 or P<0.01）. Compared with control group and si-NC group， the expression level of PAX8-AS1 mRNA in the cells in si-PAX8-AS1 group was decreased（P<0.01）， and the expression level of miR-22-3p was significantly increased（P<0.01）； compared with si-NC group，the expression level of miR-22-3p in the cells in si-PAX8-AS1+inhibitors group was increased（P<0.01）.The MTT assay results showed that compared with control group，the proliferation activities of the cells in si-PAX8-AS1 group，si-PAX8-AS1+inhibitors NC group，and si-PAX8-AS1+inhibitors group were significantly decreased after culture for 48 and 72 h （P<0.01）； compared with si-NC group， the proliferation activities of the cells in si-PAX8-AS1+ inhibitors NC group were significantly decreased after culture for 48 and 72 h （P<0.01）.The flow cytometry results showed that compared with control group， the apoptotic rate of cells in si-PAX8-AS1 group was significantly increased （P<0.01）；compared with si-NC group，the apoptotic rate of cells in si-PAX8-AS1+ inhibitors NC group was significantly increased （P<0.01）.The Transwell chamber assay results showed that compared with control group， the rates of migration and invasion cells in si-PAX8-AS1 group were significantly decreased（P<0.01）； compared with si-NC group，the rates of migration and invasion cells in si-PAX8-AS1 group and si-PAX8-AS1+ inhibitors NC group were significantly decreased（P<0.01）.The Western blotting results showed that compared with control group and NC group， the expression levels of Bax and cleaved Caspase-3 proteins in the cells in si-PAX8-AS1 group were significantly increased（P<0.01）， and expression level of the Bcl-2 protein was significantly decreased（P<0.05）.The TargetScan predicted that there were target binding sites between PAX8-AS1 and miR22-3p.The dual luciferase reporter gene assay results showed that compared with the cells co-transfected with mimics NC， the luciferase activity in the PAX8-AS1-WT cells was significantly decreased after co-transfected with miR-22-3p mimics （P<0.01）. Conclusion The PAX8-AS1 is highly expressed in the human CRC tissue， and silencing the PAX8-AS1 expression can inhibit the proliferation， migration and invasion of the CRC cells and induce the apoptosis，and its mechanism may be ralated to up-regulation the miR-22-3p expression.

Objective To discuss the effect of hypoxia inducible factor-1α （HIF-1α）/ reactive oxygen species （ROS） on the apoptosis and invasion of the non-small cell lung cancer A549 cells，and to clarify the related mechanism. Methods The A549 cells were cultured under hypoxia condition for 12， 24， 48 and 72 h to construct the hypoxia cell model. The expression levels of HIF-1α mRNA in the cells at different treatment time were detected by real-time fluorescence quantitative PCR（RT-qPCR） method， and the proliferation abilities of the cells were detected by CCK-8 assay；the hypoxia cell model was verified， and the optimal induction time was determined.The A549 cells were divided into control group（21%O₂）， model group（hypoxia induction）， NAC group（20 mmol·L^－1 NAC）， NAC+YC-1 group（20 mmol·L^－1 NAC+100 μmol·L^－1 YC-1） and YC-1 （100 μmol·L^－1 YC-1）group. The expression levels of HIF-1α and FPR1 mRNA and proteins were detected by RT-qPCR and Western blotting methods； the ROS levels were detected by flow cytometry； the autophagosome structure in the cells was observed by transmission electron microscope； the expressions of microtubule-associated protein light china 3 Ⅱ（LC3-Ⅱ） in the cells in various groups were detected by immunofluorescence； the apoptotic rates of the cells in various groups were detected by flow cytometry； and the number of invasion cells was detected by Transwell chamber assay. Results With the prolongation of hypoxia induction time， the HIF-1α mRNA expression levels in the A549 cells and cell proliferation abilities were increased（P<0.05 or P<0.01）， and the optimal hypoxia induction time was 24 h. The results of RT-qPCR and Western blotting methods showed that compared with control group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in model group were significantly increased（P<0.01）； compared with model group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased（P<0.01）.Compared with control group，the level of ROS in the cells in model group was significantly increased （P<0.01）； compared with model group，the ROS levels in the cells in NAC group， NAC+YC-1 group， and YC-1 group were significantly decreased（P<0.01）；compared with NAC group，the ROS level in the cells in NAC+YC-1 group was significantly decreased（P<0.01）.The transmission electron microscope results showed that the organelle structure of the cells in control group was intact and no autophagosome structure was seen； a large number of autophagosomes were seen in the cells in model group， obvious vacuoles were visible， and intracellular organelles were degraded； compared with model group， the number of autophagosomes in the cells in NAC group， NAC+YC-1 group and YC-1 group were decreased，among them the autophagosomes in the cells in NAC+YC-1 group were the least. The immunofluorescence assay results showed that the expression intensity of LC3-Ⅱ in the cells in model group was significantly increased compared with control group， and the expression intensities of LC3-Ⅱ in the cells in NAC group， NAC+YC-1 and YC-1 group were signficantly decreased compared with model group.Compared with control group， the apoptotic rate of the cells in model group was significantly decreased（P<0.01）； compared with model group， the apoptotic rates of the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly increased（P<0.01）； compared with NAC group，the apoptotic rate of the cells in NAC+YC-1 group was increased（P<0.01）.The results of Transwell chamber assay showed that compared with control group the number of the invasion cells in model group was significantly increased（P<0.01）； compared with model group， the numbers of invasion cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased （P<0.01）；compared with NAC group，the number of the in vasion cells in NAC+YC-1 group was decreased（P<0.01）. Conclusion Inhibition of HIF-1α/ROS under hypoxia condition can promote the apoptosis and inhibit cell invasion of cancer cells，and its mechanism may be realted to inhibiting the autophagy of lung cancer cells.

Objective To construct a cDNA library by using mRNA from Gloydius ussuriensis(G. ussuriensis) venom gland, to clone and analyze serine proteinase gene from the cDNA library. Methods Total RNA was isolated from venom gland of G. ussuriensis, mRNA was purified by using mRNA isolation Kit. The whole length cDNA was synthesized by means of smart cDNA synthesis strategy, and amplified by long distance PCR procedure, lately cDAN was cloned into vector pBluescript-sk. The recombinant cDNA was transformed into E.coli DH5α. The cDNA of serine proteinase gene in the venom gland of G. ussuriensis was detected and amplified using the in situ hybridization. The cDNA fragment was inserted into pGEMT vector, cloned and its nucleotide sequence was determined. Results The capacity of cDNA library of venom gland was above 2.3×10⁶. Its open reading frame was composed of 702 nucleotides and coded a protein pre-zymogen of 234 amino acids. It contained 12 cysteine residues. The sequence analysis indicated that the deduced amino acid sequence of the cDNA fragment shared high identity with the thrombin-like enzyme genes of other snakes in the GenBank. The query sequence exhibited strong amino acid sequence homology of 85% to the serine proteas of T. gramineus, thrombin-like serine proteinase Ⅰ of D. acutus and serine protease catroxase Ⅱ of C. atrox respectively. Based on the amino acid sequences of other thrombin-like enzymes, the catalytic residues and disulfide bridges of this thrombin-like enzyme were deduced as follows: catalytic residues, His⁴¹,Asp⁸⁶, Ser¹⁸⁰; and six disulfide bridges Cys⁷ -Cys¹³⁹, Cys¹²⁶ -Cys⁴², Cys⁷⁴ -Cys²³², Cys¹¹⁸ -Cys¹⁸⁶, Cys¹⁵⁰-Cys¹⁶⁵, Cys¹⁷⁶-Cys²⁰¹. Conclusion The capacity of cDNA library of venom gland is above 2.3×10⁶, overtop the level of 10⁵ capicity, The constructed cDNA library of G. ussuriensis venom gland would be helpful platform to detect new target genes and further gene manipulate. The cloned serine proteinase gene exhibits strong amino acid sequence homology of 85% to the serine proteas of T. gramineus, thrombin-like serine proteinase Ⅰ of D. acutus and serine protease catroxase Ⅱ of C. atrox respectively.

Abstract:Objective To construct the vector for β-galactosidase (β-gal) gene expression regulation with tetracycline-regulated gene expression system,and to control β-galactosidase gene expression in hepG2 cells with the vector.So that offer a experimental base for regulating gene expression for liver cancer gene therpy with this system.Methods The primers for explanding two tetracycline operator (TetO2) gene according to the gene nucleotide sequences of TetO2 gene and pcDNA3.1 vector were designed.TetO2 gene was amplified by PCR with pcDNA3.1 plasmid as template.The TetO2 PCR products were cloned into pcDNA3.1 and the vector was named as pcDNA3.1-TetO2.The pcDNA3.1-Teto2 with TetO2 PCR products sequence was analyzed by ABI3130 sequencing analysis.Then β-gal gene was cloned into pcDNA3.1-TetO2,the vector was named as pcDNA3.1-TetO2-β-gal.The pcDNA6/TR plasmid vector with tetracycline repressor(TR) was transfected into HepG2 cells by Lipotap,and the stable transfection cells were screened by blasticidin.TR gene expression was detected by RT-PCR in HepG-2 cells with and without pcDNA6/TR plasmid transfection.The pcDNA3.1-TetO2-β-gal plasmid vector was transfected into HepG2 cells with and without pcDNA6/TR plasmid transfection,and 3 to 4 d later, these transfected gene cells were treated with doxycline（4 mg•L^-1）.After 48 h,β-gal gene expression was detected with β-gal cell staining.Results The pcDNA3.1-TetO2 sequencing analysis results showed that a cassette was made for a cytomegalovirus-type 2 tetracycline operator (TetO2)-TetO2 promoter in pcDNA3.1-Teto2.The double digestion reslut of pcDNA3.1-TetO2-β-gal plasmid vector demonstrated that β-gal gene was successfully cloned into pcDNA3.1-TetO2 vector .The RT-PCR result of TR gene showed TR gene could be expressed in HepG2 cells with pcDNA6/TR plasmid transfection and TR gene didn’t express in HepG2 cells without pcDNA6/TR plasmid transfection.β-gal gene expressed in HepG2 cells without pcDNA6/TR plasmid transfection,but didn’t express in HepG2 cells with pcDNA6/TR plasmid transfection.However,β-gal gene expression could be induced with doxycline in HepG2 cells with pcDNA6/TR plasmid transfection. Conclusion The tetracycline-regulated gene expression system vector for regulating β-gal gene expression is successfully constructed and the vector system can control β-gal gene expression in HepG2 cells.

Abstract:Objective To observe the migration,colonization and repairing effects of marrow mesenchymal stem cells (MSCs) on thymus tissue injury induced by ionizing radiation in mice.Methods MSCs of C57BL/6 mice were isolated,purified and cultivated in vitro.Their migration and colorization were observed with laser confocal microscopy 1,5 and 10 d after DAPI labeled. MSCs were injected into the thymus tissue of mice through tail vein.The model of thymus tissue injury induced by whole-body X-irradiation was established.The mice were divided into four groups: normal,irradiation,irradiation+saline,and irradiation+MSCs groups.The apoptosis was detected by flow cytometry and the repairing effect of MSCs on thymus tissue injury was observed by histological method 3 months later.Results The occurrence of MSCs in the thymus was observed 1 d after MSCs injection,the diffusion of MSCs in the thymus appeared 5 d later,and widely dispersed 10 d later.The apoptotic rate of thymocytes in irradiation group was higher than that in normal(P<0.05) and was lower than that in MSCs group(P<0.05).The structures of cortex and medulla of thymus were clear in mice in normal group,there were a large number of lymphocytes in the cortex and small number of lymphocytes in the medulla.The structures of cortex and medulla of thymus were unclear in mice in both irradiation,irradiation and saline groups.The lymphocytes in thymus showed extensive coagulation necrosis.There were remnants or newborn lymphoid tissue in the cortex and medulla in mice in irradiation+MSCs groups. Conclusion MSCs can be rapidly enriched in thymus tissue and promote regeneration and repair of damaged thymus.

Abstract:Objective To investigate the expression and distribution of cyclooxygenase-2 (COX-2) in periodontal tissues during experimental tooth movement in rats,and  study  the relationship between  COX-2 and vesscular reconstruction in experimental tooth movement process.Methods Thirty-five healthy Wistar rats were divided randomly into 7 groups on average:normal group and experimental groups for 1,3,5,7,14 and 21 d.A NiTi coil spring with 0.294 N mesial force was connected between first molars of maxillary and the upper incisors.The histological sections were stained with goat anti rat COX-2 antibody,and computer image analysis was used to study the expression of COX-2 in the periodontal tissues of rats.Results Pressure area:compared with normal group(134.75±5.25) the COX-2 expression in 1 d group (147.73±3.27)increased (P<0.05) after tooth movement and reached the maximum at 5 d(154.32±9.54).Tension area:compared with normal group,the COX-2 expression in 3 d group (139.16±5.01)increased( P<0.05) after tooth movement and reached the maximum at  7 d(159.46±7.48).There was no significant difference of COX-2 expression in pressure area and tension area bewteen 21 d group and normal group (P>0.05).
Conclusion The expression of COX-2 in  periodontal tissues during experimental  tooth movement increase,suggesting that COX-2 can promote the vascular reconstruction in periodontal tissues during orthodontic tooth movement.

Objective To study the effects of ginseng fruit saponins（GFS）on the coronary circulation and cardial oxygen metabolism in dogs with acute myocardial infarction. Methods The acute myocardial infarction model was induced by ligating the left anterior descending coronary of dogs for 6 h. 90 min after treatment with GFS, the blood flow of coronary artery was determined by using electromagnetism flowmeter with MP-27 type. At the same time，the blood oxygen co ntents of artery and vein were determined by using analysis instrument of blood gas with CORNIHG 178 type. Results After treated by GFS (in a dosage of 5 and 10 mg·kg^-1 iv drip for 90 min)，the myocardial blood flow (MBF) was increased and coronary vascular resistance (CVR) was decreased significantly. Meanwhile the cardiac oxygen consumption and myocardial oxygen utilization rate were also decreased, but cardiac oxyg en consumption index had not changed significantly. Conclusion GFS has protective effects on acute myocardial ischemia through improving the coronary circulation, increasing myocardial blood supply and decreasing cardiac oxygen consumption and myocardial oxygen utilization rate. 

Objective To observe the protective effects of Shengmai inject on on ischemic myocardium in patients with acute myocardial infarction (AMI). Methods Forty-eight AMI patients were randomly divided into Shengmai injection treatment group (n=26) and non-Shengmai injection treatment group (n=22), and 24 normal healthy subjects were used as controls. The levels of MDA and SOD in serum were assayed with colorimetric methods. Results The levels of SOD in all AMI patients were obviously lower than that in normal healthy sujects before therapy, and the levels of MDA were significantly increased (P<0.01); as the time went on, the level of SOD was increased gradually in Shengmai injection treatment group, and MDA level was decreased gradually. Although the levels of MDA and SOD in non-Shengmai injection treatment group had the same trends with the Shengmai injection treatment group, but there was marked difference of the serum content between the two groups (P<0.01). Conclusion With the effects of anti-oxygen-derived free radicals and anti-lipid peroxidation, Shengmai injection has protective effects on the ischemic myocardium in patients with AMI.

Objective To analyze the effect of rheumatoid arthritis （RA） on the phenotypes of plasma cells of gingiva and expression of receptor activator of nuclear factor-κB ligand（RANKL） in the patients with periodontitis，and to clarify its possible mechanism. Methods Sixty subjects who came to our hospital were selected and divided into healthy group， periodontitis group and periodontitis+RA group according to inclusion criteria， and there were 20 subjects in each group.The gingival index （GI）， probing depth （PD）， bleeding on probing （BOP） and clinical attachment loss （CAL） of the subjects were recorded. The expression levels of proliferation inducing ligand（APRIL） and B lymphocyte stimulator（BLyS） in gingiva tissue of the subjects in various groups were detected by real-time fluoresence quantatitive PCR（RT-qPCR） method；the percentages of CD38+ cells， CD138+ cells in gingiva tissue of the subjects in various groups were analyzed by flow cytometry；the percentages of RANKL+ cells in the gingival plasma cells and the RANKL levels in plasma cells of gingiva of the subjects in various groups were detected by flow cytometry and enzyme-linked immunosorbent assay（ELISA） methods； the growth of osteoclasts in the subjects in various groups was observed by anti-tartrate acid phosphatase （TRAP） staining； the expression levels of RANKL， receptor activator of nuclear factor-κB（RANK） and tumor necrosis factor associated factor 6（TRAF6） mRNA in plasma cells of gingiva of the subjects in various groups were detected by RT-qPCR method. Results Compared with healthy group， GI， PD， BOP and CAL of the patients in periodontitis group and periodontitis+RA group were significantly increased （P<0.05）； compared with periodontitis group，the PD and CAL of the patients in periodontitis+RA group were increased（P<0.05）.Compared with healthy group， the expression levels of APRIL and BLyS mRNA in gingiva tissue of the patients in periodontitis group and periodontitic+PA group were increased（P<0.05）；compared with periodontitis group，the expression level of APRIL mRNA of the patients in periodontitis+RA group was increased（P<0.05）.Compared with healthy group， the percentages of CD38+ cells and CD138+ cells in gingiva tissue of the patients in periodontitis group and periodontitis+RA group were significantly increased（P<0.05）；compared with periodontitis group， the percentage of CD138+ cells in gingiva tissue of the patients in periodontitis+RA group was increased （P<0.01）.Compared with healthy group， the percentages of RANKL+ cells in plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）；compared with periodontitis group， the percentage of RANKL+ cells in the plasma cells of gingiva the patients in periodontitis+RA group was increased（P<0.05）. Compared with healthy group， the percentages of RANKL in plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）. Compared with healthy group， the number of normal induced TRAP+osteoblast-like cells of the patients in periodontitis group and periodontitis+RA group were increased（P<0.05）； compared with periodontitis group， the number of normal induced TRAP+ osteoblast-like cells of the patients in periodontitis+RA group was increased （P<0.05）； compared with the normal induced cells， the numbers of anti-RANKL induced TRAP+osteoblast-like cells in various groups were decreased（P<0.05）.Compared with healthy group，the expression levels of RANKL， RANK， and TRAF6 mRNA in the plasma cells of gingiva of the patients in periodontitis group and periodontitis+RA group were significantly increased （P<0.05）；compared with periodontitis group， the expression levels of RANKL， RANK， and TRAF6 mRNA in the plasma cells of gingiva of the patients in periodontitis+RA group were increased （P<0.05）. Conclusion The differentiation level and the ability on promoting osteoclastogenesis of plasma cells of gingival of the patients with periodontitis complicated with RA are higher than those in the patients with single periodontitis， which may be a potential cause for RA aggravating periodontal tissue destruction in the patients with periodontitis.

Objective   To explore the  relationship between glycemic control and implant healing
 in the patients with type 2 diabetes mellitus by studying the influence of diff
erent blood sugar levels in  the inflammatory indexes of implants and implant st
ability.Methods 21 patients with type 2 diabetes mellitus(diabetes group) and 10 healthy individuals (control
 group) were selected.40 implants were placed under local anesthesia, blood routine exam
ination was performed before and 24 h after operation,and the amount of inflammatory cells was counted.The glycated hemoglobin (HbAlc) levels
were assayed 8  and 12 weeks after implantation,the mean values were recorded as blood sugar indexes.The implant stability indexes were detected by means of
resonance frequency analysis on weeks 0,2,4,6,8 and 12 after implantation.The maximum decrease in
 stability from baseline and the time for stability level returned to baseline were calculated.
Results The inflammatory cell levels in diabetes group were higher than those in non-diabetic after implantation（P<0.05）,while no difference was found
before implantation（P>0.05）.The implant stabilities were compared according to HbAlc
levels;the patients with  higher HbA1c level had a greater maximum decrease in stability from baseline,while the difference between various groups had statistical significance（P<0.01）.The patients with HbA1c ≥7.6% required a longer time for stability level to returne
 to baseline than the patients with HbA1c ≤7.5%（P<0.01）.Conclusion The inflammatory reaction and implant stability in the patients with type 2 di
abetes mellitus are related to glycemic control.Better control of blood glucose will improve implant healing.

Abstract:Objective To investigate the degeneration of dopaminergic neurons induced by MPP⁺ in mouse mesencephalic dissociated culture and establish the cell model for parkinson’[KG-*3]s disease( PD) research in vitro.Methods OF1/SPF mouse embroys of 14 d were used to make mesencephalic dissociated culture,and the culture were divided into control and MPP⁺-treated groups.MPP⁺ with different final concentrations (0.1,1,10 and 15 μmol•L^-1)　 were added into the cultures on the 10th day in vitro,and incubated for 48 h.Then the dopaminergic neurons were identified by TH immunostaining.Hoechst 33342 was used to identify the apoptotic cells.Results The number of dopaminergic neurons was 875±23/well in control group,while there were 612 ±25,586±32,459±16 and 435±19 dopaminergic neurons per well in the 0.1,1,10 and 15 μmol•L^-1 MPP⁺-treated groups respectively,the numbers of dopaminergic neuron decreased significantly in MPP⁺ groups compared with control group（P<0.05）. On the 12th day in vitro,(5.45±0.29 )% cells in control group appeared as typical apoptotic appearance,a higher apoptotic rate was found in MPP⁺ group (26.97%±1.36% )(P<0.05).Conclusion All the concentrations of MPP⁺（0.1,１,10 and 15 μmol•L^-1） could induce the damage and dead of dopaminergic neurons. The degeneration and death are both dose- and time-dependent.Apoptosis may play an important role in the damage of dopaminergc neurons induced by MPP⁺.

Objective： To discuss the effect of Huaier aqueous extract （HAE） on the proliferation， migration， cell cycle，and apoptosis of tamoxifen （TAM） resistant breast cancer LCC2 cells， and to clarify its mechanism. Methods The cells were divided into control group （given DMEM basal medium） and TAM group （given 2 μmol·L^－1 TAM）， TAM+HAE group （given 2 μmol·L^－1 TAM+4 g·L^－1 HAE） and 0， 2， 4， 8， and 16 g·L^－1 HAE groups. MTT method was used to detect the survival rates and the proliferation rates of the cells in various groups； cell scratch test was used to detect the migration rates of the cells in various groups；flow cytometry was used to detect the percentages of the cells at different cell cycles and apoptotic rates of the cells in various groups；Transwell chamber test was used to detect the migration number of the cells in various groups； Western blotting method was used to detect the expression levels of estrogen receptor α （ER α） in each group，amplified in breast cancer 1（AIB1），and survivin proteins in the cells in various groups. Results The MTT assay showed that after treated for 24 h， compared with 0 g·L^－1 HAE group， the survival rates of the HAE cells in 4，8，and 16 g·L^－1 HAE groups were significantly decreased（P<0.01）. At 48 h after treatment， compared with 0 g·L^－1 HAE group， the survival rates of the cells in 2，4，8， and 16 g·L^－1 HAE groups were significantly decreased （P<0.01）；compared with 4 g·L^－1 HAE group，the survival rate of the cells in 8 g·L^－1 HAE groups was decreased（P<0.05）；compared with 24 h after treatment， the surival rates of the cells in different concentrations of HAE groups were significantly decreased （P<0.01） after treated for 48 h. At 48 treatment，compared with TAM and 4 g·L^－1 HAE group，the proliferation rate of the cells in TAM+HAE group was significantly decreased（P<0.01）.The cell scratch test results showed that the cell migration rate of the cells in 4 g·L^－1 HAE group was lower than that in 0 g·L^－1 HAE group after treated for 48 h （P<0.05）. The flow cytometry results showed that after treated for 48 h， compared with 0 g·L^－1 HAE group， the percentage of the cells at S phase and the apoptotic rate of the cells in 4 g·L^－1HAE group were significantly increased （P<0.05 or P<0.01）. The Transwell chamber experiment results showed that after treated for 48 h， compared with TAM group and 4 g·L^－1 HAE group， the migration number of the cells in TAM+HAE group was significantly decreased （P<0.01）. The Western blotting results showed that compared with MCF-7 cells，the expression levels of ERα，AIB1， and survivin proteins in the LCC2 cells were increased （P<0.05 or P<0.01）；at 48 h after treatment， compared with 0 g·L^－1 HAE group，the expression levels of ERα，AIB1， and survivin proteins in the LCC2 cells in 2，4，8， and 16 g·L^－1 HAE groups were significantly decreased （P<0.01）. Conclusion HAE can inhibit the proliferation and migration， arrest the cell cycle at S phase， and promote the apoptosis of the LCC2 cells，and it can also restore the sensitivity of the LCC2 cells to TAM， and its mechanism may be related to down-regulation of the expression levels of ER α， AIB1，and survivin proteins in the LCC2 cells.

Objective To study the absorpion and utilization of calcium gluconate and calcium lactate in rats in vivo. Methods Weaning male Wistar rats were randomly divided into 5 groups and fed on CaCO₃, skimmed milk powder, calcium gluconate, calcium lactate， and basic low calcium meal in semi-purified diets, respectively, for 4 weeks. Meanwhile，the experiments of 3 days′ feed efficiency and 4 days′ calcium metabolism were carried out on the 20th and 25th day, respectively. Results In calcium-gluconate group, the incremental rate of body weight,the absorption and retention rate of calcium,the femur index, and retention rate of calcium in femur were 92.1%,68.2%,85.1%,4.96, and 4.6%, respectively, all indexes were lower than those in milk-powder group and higher than those in CaCO₃ group (P<0.05 or P<0.01). The items mentioned above in calcium-lactate group were 75.1%,65.5%,82.0%,4.88 and 4.1%, respectively,also higher than those in CaCO₃ group (P<0.05 or P<0.01),no differences were found between calcium-gluconate group and calcium-lactate group except that the incremental rate of body weight in calcium-lactate group was lower than that in calcium-gluconate group.Conclusion Both calcium gluconate and calcium lactate can be served as the good sources for dietary calcium supplement.

Objective To discuss the feasibility and effect of labeled antibody on targeting therapy of systemic lupus erythematosus (SLE) associated thrombocytopenia. Methods MTX-IVIG conjugate was prepared by indirect and direct cross linking,and the binding ability of it to Fc fragment was detected by indirect immunofluor-escence.The killing activity of the conjugate was detected by MTT method using murine macrophage with Fc receptor and strain U937 of human monocytic leukemia cell as targets. Results Conjugation showed stronger cytotoxicity upon target cells than free MTX,and it showed less cytotoxic effect on Fc receptor negative cells compared with the positive ones. IVIG-HSA-MTX restrained the phagocyte of macrophage. The killing effect of IVIG-HSA-MTX was significantly stronger than that of IVIG-MTX. Conclusion The conjugation can show a highly specific cytotoxicity upon mononuclear-macrophage in vitro.

Objective To discuss the clinical efficacy of minimally invasive circumcision， traditional open surgery， and combination surgery in the treatment of multiple breast tumors complicated with larger lesions， and to provide the clinical the basis for the treatment of the patients with multiple breast tumors. Methods A total of 167 patients with multiple breast tumors were divided into minimally invasive surgery group （n=83， treated with minimally invasive circumcision）， traditional surgery group （n=42， treated with traditional open surgery）， and combination surgery group （n=42， treated with minimally invasive circumcision and traditional open surgery）. The surgical time， intraoperative bleeding vomule， healing time of the incision， ratios of incision number to tumor number， and postoperative incidences of ecchymosis， infection， hematoma， obvious scar， and residual lesion，and the satisfaction rates of the patients in three groups were observed. Results Compared with traditional surgery group， the ages of the patients in minimally invasive surgery group and combination surgery group were younger （P<0.01）. Compared with traditional surgery group and minimally invasive surgery group，the surgical time of the patients in combination surgery group were significantly increased （P<0.01）；compared with traditional surgery group， the intraoperative bleeding vomule of the patients in combination surgery group was decreased （P<0.01）；compared with minimally invasive group， and the intraoperative bleeding vomule of the patients in combination surgery group was increased （P<0.01）.Compared with traditional surgery group and minimally invasive surgery group， the ratio of incision number to tumor number of the patients in combination surgery group were decreased significantly （P<0.01）； compared with minimally invasive surgery group，the wound healing time of the patients in combination surgery group was decreased significantly （P<0.01）. There were no statistically significant differences in the incidences of total postoperative complications， hematoma， ecchymosis， obvious scars， infection， and residual complications of the patients among three groups （P>0.05）. Compared with traditional surgery group， the psychological satisfaction rate of the patients in combination surgery group was increased （P<0.05）. Conclusion The minimally invasive breast circumcision combined with traditional open surgery have a good therapeutic effect in the treatment of the patients with multiple breast tumors complicated with large lesions，with fewer surgical incisions and high satisfaction rates；it is suitable for the promotion and application in the specific populations.

Abstract:Objective To establish quantifiable primary and experimental lung metastasis  mouse models by using GFP labeled MDA-MB-231 cell lines and provide basis for the study on metastasis mechanism of breast cancer.  Methods The MDA-MB-231 cells were infected by GFP labeling lentiviral vector-mediated packaging system and inoculated subcutaneously into Balb/C nude mice or through tail vain into SCID mice to set up primary and experimental lung metastasis models.Then the mice were executed at 8 th and 5 th weeks,and the  lungs were  extracted and analyzed by stereomicroscope.  Results 8 weeks after the cells were inoculated in primary lung metastasis mice,the tumor in situ was observed with the diameter of 15 mm. The metastasis foci in the lung were invisible to naked eye,but were visible at 488 nm excitation wavelength with green fluorescence. 5 weeks after the cells were injected in experimental lung metastasis mice,the metastasis foci were still invisible to naked eye,but were clearly visible at 488 nm excitation wavelength with green fluorescence,which were easy to be observed and calculated.  Conclusion The mouse model for lung metastasis using MDA-MB-231 cells with countable green fluorescence foci is successfully established.

Abstract:Objective To explore the feasibility of orthotopic transplantion tumor model of hepatoma in mice and evaluate the biological features.Methods 5×10⁶ H22 cells were inoculated into the liver of mice after anesthetizing.The liver,spleen,lung and kidney were taken from the mice 13 d after inoculation(n=8) or from the died mice(n=9) for morphological evaluation.Proliferating cell nuclear antigen(PCNA) and factor Ⅷ related antigen(FⅧRag) were analyzed with immunohistochemical staining.Results The average survival time of the mice was 18.8 d.Until the 13th day after inoculation,the successful rate of orthotopic transplantation tumor (model of liver cancer) was 100%(8/8),spontaneous lung metastatic rate was 37.5%(3/8).Fifty percent of mice(4/8) produced ascites,and the average amount of ascites was （6.25±3.67）mL.The average size of the tumor was （0.35±0.14） cm³.In the natural died mice,spontaneous lung metastatic rate was 100%（9/9）,100% of mice (9/9)produced ascites. The average volume of acites was （10.88±1.92）mL,which was evidently more than that of the 13th day（P<0.01）.The average size of the tumor was (0.61±0.31) cm³, which was significantly bigger than that of the 13th day（P<0.05）.When the mice died,most of tumor nodules were single and the cellular 〖JP2〗atypia was apparent.The proliferation index and microvessel density of tumor tissue were 71.39%±〖JP〗20.74% and (39.25±13.71)/400 high magnification respectively.Conclusion The successful rate of orthotopic transplantation tumor model of liver cancer is high.The tumor possess high lung metastatic rate and plenty of microvessels and strong proliferative ability.

Objective To evaluate the efficacy and safety of a combination use of clobetasol propionate and retin-A cream in the treatment of psoriasis vulgaris. Methods A randomized controlled and double blind trial was used. 72 patients were divided into trial group（combination of clobetasol propionate and retin-A cream）,control group 1（Diwei cream）,and control group 2（Enfu cream）. The efficacy and safety were observed. Results The cure rate and efficacy rate were 58.3% (14/24) and 100.0% (24/24),respectively, for trial group whereas cure rate and efficacy rate were 41.7% (10/24) and 75.0% (18/24), respectively, for control group 1 and 16.7% (14/24) and 75.0% (18/24) for control group 2. The difference was significant between various groups (P<0.05). There were no local and general adverse effects of drugs in all groups. Conclusion A combination use of clobetasol propionate and retin-A cream is effective and safe in treatment of psoriasis vulgaris.

Objective To investigate the effect of propofol on the expression of TGF-β₁ in cerebral cortex during focal cerebral ischemia-reperfusion in rats. Methods The focal cerebral ischemia-reperfusion model was established by thread embolism of middle cerebral artery. Forty-five male Wistar rats were randomly allocated to sham-operation group (n=5), ischemia-reperfusion group (n=20) and propofol group (n=20), rats were subjected to 2 h of focal ischemia by left middle cerebral artery occlusion and then reperfused. At 3, 6, 24, and 72 h after reperfusion the rats were decapitated, the expressions of TGF-β₁ in ischemic cortex were tested by immunohistochemical method. At 24 h after reperfusion, scores of neurological deficit were estimated and the change of histomorphology in ischemic cortex was observed under the light microscope by using HE staining. Results The expression of TGF-β₁ increased significantly after focal cerebral ischemia-reperfusion, reached to maximal expression at 24 h after reperfusion (P<0.01), and decreased to normal at 72 h after reperfusion. The scores of neurological deficit in rats increased significant ly. The followings were observed in histomorphology: congestion and edema in ischemic cort ex, degeneration and necrosis of neuron. Expression of TGF-β₁ in pro pofol gr oup increased significantly than that in ischemia-reperfusion group (P<0.05). The scores of neurological deficit and the damage of histomorphology in ischemic cortex lowered than that in propofol group. Conclusion The results show that propofol can stimulate the expression of TGF-β₁ to produce neuroprotection. Key words：propofol/pharmacology; cerebral ischemia; reperfusi on injury; transforming growth factor beta; disease models,animal

Abstract:Objective To explore the molecular mechanism of TGFβ in the progress of breast cancer by examining the expressions of TGFβ₁,TGFβ receptor Ⅱ and Smad transcriptional factors in normal human breast epithelium cells and MCF7 cells. Methods RT-PCR was used to measure the expression levels of TGFβ₁,TGFβ receptor Ⅱ,Smad4,Smad6 and Smad7 in normal human breast epithelium cells and MCF7 cells. Results There were no any significant differences of the ratios of TGFβ₁,TGFβ receptor Ⅱ,Smad6,Smad7 to GAPDH between normal human breast epithelium cells （0.721±0.214,1.001±0.312,0.689±0.12,0.221±0.124）and MCF7 cells （0.698±0.324,0.998±0.217,0.718±0.257,0.246±0.138）while the expression of Smad4 in MCF7 cells was significantly lower than that in normal human breast epithelium cells (P<0.05). Conclusion The low expression of Smad4 in MCF7 cells may be associated with the promotion effect of TGFβ on breast cancer.

Objective To study the action of corn active polysaccharide (CAP) on blood sugar levels in diabetic mice.Methods Fifty mice were randomly divided into control group, alloxan model group, CAP 3.50,1.75, 0.88 g·kg^-1 groups (n=10). Mice in each group were exhibited with CAP once a day for 15 d，on the sixth day,mice in every group were injected with alloxan physiological salt solution 80 mg·kg^-1 in caudal vena except control group, mice in control group were injected with physiological salt solution at the same volume. 16 h before the last exhibition, mice in every group were starved. 1 h before the last exhibition, blood was extracted in eyeballs, sera were separated and the contents of blood sugar were determined by auto-biochemistry analyzer, mice were killed at once. The liver samples were obtained and the contents of liver glycogen were determined. Another 60 mice were divided into 5 groups with the method men tioned above. Each group was exhibited once a day for 10 d continuously. 1 h after the last exhibition, except control group, the mice were injected with adrenalin hydrochloride 250 μg·kg^-1 through abdominal cavity, after30 min, blood was extracted by decapitation, and the contents of blood sugar and liver glycogen were determined by auto-biochemistry analyzer.Results CAP 3.50　g·kg^-1 had obvious inhibitory effect on the raise of blood sugar in diabetic mice （model group）（P＜0.05）, it had obvious action on reducing the contents of liver glycogen in diabetes mice（model group）（P＜0.05）,and 3.50　g·kg^-1 was superior to 1.75 g·kg^-1. CAP with different doses had obvious actions on inhibiting the raise of blood sugar in adrenine mice（model group）（P＜0.001, P＜0.01, P＜0.05）,and had obvious action on increasing the reduction of liver glycogen on adrenine mice（model group）,and 3.50 g·kg^-1 was superior to 1.75 g·kg^-1. Conclusion CAP has the function of lowering blood sugar levels in alloxan diabetic mice and hyperglycemia mice induced by adrenine.

Objective To investigate the effects of ginsenoside Rg₂ on hemodynamics and oxygen metabolism in narco-canine with acute cardiogenic shock. Methods Model of acute cardiogenic shock was made by ligating left anterior descending branch of coronary artery(LAD). Hemodynamics and oxygen metabolism in narco-canine were observed after administrated with 2 and 1 mg•kg^-1 ginsenoside Rg₂. Results Ginsenoside Rg₂ increased mean blood pressure (MBP),left ventricular systolic pressure (LVSP),±dp/dt_max ,and cardiac output (CO), remarkably, and reduced the exponent of oxygen consuming of myocardium and the rate of oxygen intake of myocardium compared with control group. Conclusion Ginsenoside Rg₂ has therapeutical effect on acute cardiogenic shock in canine.

Objective To investigate the anti-inflammatory effects of Hushigutong Granule. Methods Fifty Wistar rats were randomly divided into control group,aspirin group, and three groups of Hushigutong Granule at different doses (15,30,and 60 mg•kg^-1). Anti-inflammatory effects of Hushigutong Granule were carried through inflammatory models of auris swell induced by dimethylbenzene in mice, of foot swell induced by egg white and carageen glue in rats, of gasbag swell induced by croton oil in rats and penetration of capillary vessel. Results Hushigutong Granule (30 and 60 mg•kg^-1) had significant inhibitory action on auris swell induced by dimethylbenzene in mice, foot swell induced by egg white and carageen glue, gasbag swell induced by croton oil and penetration of capillary vessel in rats. There were significant differences compared with control group (P<0.05 or P<0.01). Conclusion Hushigutong Granule has significant inhibitory effects on many experimental inflammatory models.

To screen one kind of antitumor immunoadjuvant that could enhance the anti-tumor activity of heat shock fusion protein(HSP-MUC1).Methods MUC1-positive tumor bearing mice were subcutaneously injected by HSP-MUC1 combined with a novel CpG ODN or hIFNα2b,four groups were set up respectively as PBS,HSP-MUC1,CpG ODN(IFNα2b) and HSP-MUC1+CpG ODN(HSP-MUC1+IFNα2b).The tumor incidence and tumor volume in various groups were observed and calculated. Results The tumor incidence and tumor weight of mice in HSP-MUC1+CpG ODN group were lower than those in PBS group and HSP-MUC1 group(P<0.05); CpG ODN alone also displayed the similar anti-tumor effect as HSP-MUC1+CpG ODN,it could decrease the tumor incidence and tumor weight of mice compared with PBS and HSP-MUC1(P<0.05); And the tumor incidence and tumor weight of mice in HSP-MUC1+hIFNα2b group had no difference with those in PBS group and HSP-MUC1 group(P> 0.05). Conclusion CpG ODN could enhance the anti-tumor activity of HSP-MUC1 but need to be further determined for its optimal injection time；and human IFNα2b could not enhance the anti-tumor activity of HSP-MUC1.

Objective To investigate the preliminary effects and mechanism of oxymatrine(OMT) on analgesia. Methods Mice were randomly divided into 6 groups:normal saline group, OMT groups with different doses (50,100,and 200 mg·kg^-1), pethidine hydrochloride (10 mg·kg^-1)group, and N^G-nitro-L-arginine methyl ester (L-NAME) 20 mg·kg^-1+OMT 100 mg·kg^-1 group. The number of writhing within 15 min was observed in pain mouse models caused by acetic acid. Femal e mice were randomly divided into 6 groups as the methods mentioned above. Micewere placed on the hot plate maintained at (55.0±0.5)°C and the latency of licking of the hind paws was observed. Results The number of writhing was decreased and the latency of licking of the hind paws was prolonged in a dose-pendent manner after intraperitoneal injection of OMT 50,100, and 200 mg·kg^-1 in mice(P<0.05，P<0.01，P<0.001). L-NAME injected in advance enhanced the effect of OMT on antagonism of the writhing response caused by acetic acid.Conclusion These results suggest that OMT can antagonize the acute pain caused by acetic acid and hot plate in a dose-dependent manner in mice. The analgesic action of OMT may be not mediated completely by NO-cGMP pathway.

Objective To investigate the expression of tight junctions protein claudin-6 in breast cancer cell lines and tissues and its relationship with metastasis.Methods RT-PCR and immunohistochemistry analysis were used to detect claudin-6 mRNA and protein expressions in human mammary cancer tissues and cell lines. Results claudin-6 gene was expressed in mammary gland epithelial cell HBL-100，but undetectable in other cell lines；compared with benign mammary gland,fibroadenoma，claudin-6 was down-regulated in breast cancer（P<0.05）.Furthermore，the down -regulation had significantly negative correlation with mammary tumor pathological grades and metastasis（P<0.05）,but not related to age and clinical stages （P＞0.05）.Conclusion claudin-6 may play a negative regulatory effect on mammary carcinogenesis and metastasis,and may be a potential indicator for mammary cancer metastasis and prognosis.

Abstract:Objective To investigate the protective effect of Schizandrae Lignanoid(SCL）on the apoptosis of PC12 cells induced by  H₂O₂ and its relative mechanisms.  Methods PC12 cells were divided into four groups： control group,model group,high dose SCL (SCL₁) group， and low dose SCL(SCL₂) group.Apoptosis of PC12 cells was induced by  H₂O₂.The cell activity was determined by MTT,the cell apoptotic rate was determined by Annexin V-FITC/PI and ΔΨm was detected by flow cytometry.The expressions of bcl-2 and bax were detected by immunohistochemistry. Results Compared with model group，SCL increased  the survival rate of PC12 cells (P<0.01),decreased the apoptotic rate of PC12 cells  (P<0.01) and protein expression of bax (P<0.05),and increased the △Ψm  (P<0.01) and protein expression of bcl-2 significantly (P<0.05)  in a dose-dependent manner.Conclusion SCL can inhibit the apoptosis of PC12  cells induced by H₂O₂ ,which might be correlated with the inhibition of apoptosis in the path way of mitochondrion.Co

Objective To investigate the effects of oxymatrine (OMT) on blood pressure in rats. Methods Fourty-eight Wistar rats were randomly divided into 6 groups (n=8):normal saline group,OMT groups with different doses(30,60,and 90 mg•kg^-1),propranolol+OMT 60 mg•kg^-1 group and doxazosin+OMT 60 mg•kg^-1 group. Cannula was cannulated commonly into carotid artery and connected with BL-420 BioLab System to record the blood pressure and heart rate(HR) in rats. Results Mean arterial pressure(MAP) were significantly lower than control in dose-dependent manner after intraventous adminstration of OMT(30，60，and 90 mg•kg^-1). Conclusion OMT can decrease the MAP in dose-dependent manner. OMT and doxazosin have synergetic effects on MAP.

Objective To study the relationships between plasma adrenomedullin (ADM) and normal pregnancy and pathogenesis of pregnancy-induced hypertension(PIH) . Methods ADM concentrations in the plasma from 10 normal non-pregnant women,36 normal pregnant women (12 first,12 second,12 third trimester, respectively) and 30 cases of PIH (10 mild,10 moderate,10 severe, respectively) were determined by radioimmunoassay, and data were analyzed statistically. Results ADM concentrations in the first, second and third trimester of normal pregnancy increased significantly than that of normal non-pregnant women (P<0.05). ADM concentration in the plasma of patierts with PIH was higher than that of third trimester pregnancy (P<0.01). There were significant differences between mild, moderate, and severe PIH groups (P<0.05). In the PIH groups, significant positive correlation was found between plasma ADM concentration and mean arterial pressure(r=0.822,P<0.05). Incidence of low birth weight infants was related to serious degree of PIH. Conclusion ADM may involve in maintaining normal human pregnancy. ADM may increase compensatorily in the pathogenesis of PIH.

Objective: To discuss the diagnosis and treatment process of abnormally elevated blood pressure in the patient treated with rifampicin and antihypertensive drugs, to analyze the interaction between rifampicin and antihypertensive drugs, and to improve the clinicians' understanding of the administration in the patients.Methods: The clinical materials of a patient treated with rifampicin and antihypertensive drugs were collected,and the relationship between the blood pressure change and drug was analyzed.The changes of concentrations of dihydropyridine-based calcium antagonists after administration of rifampin were observed and the relative literatures were reviewed.Results: A 58-year-old man with coughing and coughing for 1 month was admitted to hospital.The patient was definitely diagnosed as tuberculosis and hypertension before admission;the patient was treated with rifampicin, isoniazid, ethambutol, pyrazinamide, and felodipine together.The original treatment plan was continued and the blood pressure of the patient was monitored.On the 9th day of anti-tuberculosis treatment, the patient developed dizziness, chest tightness, and severe fluctuations in blood pressure.Then rifampicin was stopped and antihypertensive drugs were adjusted.At the beginning of blood pressure fluctuation of the patient, the combination of angiotensin-converting enzyme inhibitors and the increasing dose of dihydropyridine-based calcium antagonists did not control the blood pressure.The blood pressure began to decrease significantly at 36 h after rifampin was stopped.On the 18th day of anti-tuberculosis treatment, the original antihypertensive plan was restored and the blood pressure remained stable.Conclusion: Rifampicin can sometimes significantly reduce the effect iveness of antihypertensive drugs (such as dihydropyridine calcium antagonists), and the clinicians should pay attention to it.

Abstract:Objective To study the changes of hematopoietic growth factors secreted by human mesenchymal stem cells from bone marrow(BM-MSC) pretreated with low dose radiation （LDR）.Methods The cultured P4 and P5 BM- MSCs were exposed to X rays atthe doses of 50 ，75 and 100 mGy (dose rate 12.5 mGy•min^-1).The changes of levels of stem cell factor (SCF), IL-6, macrophage colony-stimulating factor( M-CSF) secreted by BM-MSCs pretreated with LDR were determined by ELISA method. Results As compared with control group at the same time, the levels of SCF in experimental group had a tendency of increasing after 24 h and 48 h radiation,but only in 75 mGy group the SCF level was obviously increased (P<0.05). The levels of IL-6 in 50 and 75 mGy groups at 24 h and 48 h ,in 100 mGy group at 24 h were obviously increased compared with control group(P<0.05) .The levels of M-CSF in all the groups at 24 h, 48 h and 72 h except for the 50 mGy dose at 72 h were also increased(P<0.05),it increased markedly in 75 mGy dose group at 72 h.Conclusion LDR has hormesis effect on BM-MSCs .After LDR,the BM-MSCs grow faster and in a certain phase the expression levels of hematopoietic growth factors are increased.

Objective To study the changes of serum vascular endothelial growth factor (VEGF) in patients with breast cancer and its significance. Methods The preoperative fasting serum VEGF levels were determined in 36 patients with breast cancer, 22 patients with breast fibroadenoma and breast cystic hyperplasia， and 35 healthy female controls by using ELISA. The relationships between VEGF and clinical indexes were analyzed. Results The serum VEGF level (ng•L^-1) in breast cancer group （145.8±13.6） was significantly higher than those in benign breast disease group （80.47±3.58） and control group （58.38±7.04） （P<0.05）. The VEGF level (ng•L^-1) in patients with lymph node metastasis （188.40±19.80） was higher than that in patients without lymph node metastasis （94.02±12.04）（P<0.01）. Conclusion The serum VEGF level has certain value to make differential diagnosis between malignant and benign breast diseases and predict the possibilities of prognosis and lymph node metastasis.

Objective To investigate the relationship between the protein expression and the methylation in promoter region of death-associated protein kinase DAPK gene and oral squamous cell carcinoma (OSCC) and study the mechanism of the DAPK protein expression changes and the gene therapy method in OSCC.Methods The methylation rates in promoter region of DAPK gene in 23 cases of normal oral mucous membrane and 53 cases of OSCC tissues were detected by methylation specific PCR,and the expression rates of DAPK protein were analyzed by immunohistochemistry.Results There was no methylation in promoter region of DAPK gene in normal oral mucous membrane.The exhaustive methylation in promoter region of DAPK gene in 30 cases（56.60%）and partial methylation in 8 cases （15.09%）in OSCC were detected.The total methylation rate was 71.69% in OSCC,there was significant difference compared with normal oral mucous membrane（P<0.01）.The expression of DAPK protein was positive in 23 cases (100%) of normal oral mucous membrane and was down regulated in 36 cases (67.92%) of OSCC tissues,there was significant difference（P<0.01）.There was a significant difference in the methylation rate between down-regulated expression group in OSCC tissues（32/36,88.89%） and normal expression group（6/17,35.29%）.（P<0.01）.Conclusion The DNA methylation in promoter region might be one of the mechanisms that contribute to down-regulation of the expression of DAPK protein.The loss of DAPK gene’s function caused by the methylation in promoter region may relate to the tumorigenesis of OSCC.

To study the inhibitory effects of Fe³⁺-CMC on the expressions of transforming growth factor β(TGF-β) and fibroblast growth factor (FGF) of injured parts of postoperative peritoneal. Methods Sixty Wistar rat models with abdominal adhesion were randomly divided into control group and expriment group.At the 1st,3rd,5th,7th,14th day and 2 months later,5 rats were selected respectively in two groups and killed.The adhesion tissues were gained and the expressions of TGF-β and FGF were observed using immunohistochemistry technique. Results The expressions of TGF-β and FGF in expriment group at the 3rd,5th,7th and 14th day after operation were lower than those in control group at the same time (P<0.05). ConclusionFe³⁺-CMC can inhibit the expressions of TGF-β and FGF of injured parts of postoperative peritoneal,then decrease the postoperation adhesion.

Objective To analyze mitochondrial DNA polymorphism of district of Lubunour at the Bronze Age in Xinjiang. Methods MtDNA was extracted from 10 ancient individual bones at the tombs of Lubunour. Through four overlapping primers, mtDNA HVS-1 was amplified and sequenced. Results 10 nucleotide sequences of 363 bp length were obtained. Phylogenetic analysis indicated that the nucleotide diversity and mean pairwise differences of ancient people in Lubunour was close to European population. Conclusion An ancient European population had existed prior to 3800 years ago.

Abstract:Objective To investigate the protective effect of Schizandrae Lignanoid (SCL) on myocardial ischemia reperfusion injury in hyperlipidemic rats and its mechanisms and provide theoretical basis and experiment foundation for treatment of myocardial ischemia complicated with hyperlipidemic disease with SCL in the future. Methods After hyperlipidemic rat models were set upby oral administration of high lipid emulsion,the rats were divided into seven groups: control group，sham group，model group,SCL 60，20，5 mg•kg^-1 groups and 200 mg•kg^-1 CDT group.After reperfusion,the blood samples taken from the ventricles were assayed for blood lipid;MPO activities of ischemic myocardium were measured;Infarct area of left ventricles was measured by Evens blue-TTC staining,and myocardial pathological changes were also observed.Results Compared with model group, the myocardial infarct area/ventricle area ratios and MPO activities in SCL 60，20，5 mg•kg^-1 groups decreased (P<0.01) with the raise of SCL dose;In SCL 60 and 20 mg•kg^-1 groups myocardial fiber rupture and necrosis in ischemic area with less focal hemorrhage and inflammatory cell infiltration decreased;In SCL 60 and 20 mg•kg^-1 groups the levels of serum TC,TG,LDL-c,ApoB decreased and HDL-c,ApoA1 increased (P<0.05 or P<0.01）.Conclusion SCL has protective effects on myocardial ischemia reperfusion injury in hyperlipidemic rats and the mechanisms involved may be related to its regulation of blood lipid and inhibition of neutrophil infiltration.

Objective To discuss the effect of Cav 3.2 gene in the treatment of stress urinary incontinence （SUI） of the mice by pelvic floor electrical stimulation （PES）， and to clarify its related mechanism. Methods Thirty C57BL/6 mice were randomly divided into control group ［without vaginal dilation （VD） modeling］， VD group （VD modeling）， and VD+PES group （VD modeling combined with PES） according to the intervention measures， and there were 10 mice in each group. The mice were divided into WT-VD group （wild-type VD modeling）， WT-VD+PES group （wild-type VD modeling combined with PES）， KO-VD group （Cav 3.2 knockout type VD modeling）， and KO-VD+PES group （Cav 3.2 knockout type VD modeling combined with PES） according to the genotype and intervention mesures. Masson staining was used to observe the deposition and expression levels of collagen type Ⅰ（Col Ⅰ） and collagen type Ⅲ （Col Ⅲ） collagen fibers in urethral and anterior vaginal wall tissue of the mice in various groups. The maximum bladder capacity（MBC） and leakage point pressure （LPP） of the mice in various groups were detected， and the expression levels of integrins β1， calpain 2， Col Ⅰ， and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in various groups were detected by Western blotting method. Results Compared with VD group， the expression levels of Col Ⅰ and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in VD+PES group mice were significantly increased （P<0.01）； compared with WT-VD group， the expression levels of Col Ⅰ and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were significantly increased （P<0.01）； compared with WT-VD+PES group， the expression levels of Col I and Col Ⅲ collagen fibers in urethral and anterior vaginal wall tissue of the mice in KO-VD and KO-VD+PES groups were significantly decreased （P<0.01）. Compared with control group， the MBC and LPP of the mice in VD group were decreased （P<0.05）； compared with VD group， the MBC and LPP of the mice in VD+PES group were increased （P<0.05）. The Western blotting results showed that compared with VD group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in VD+PES group were significantly increased （P<0.01）； compared with WT-VD group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were increased （P<0.01）； compared with WT-VD+PES group， the expression levels of Col Ⅰ and Col Ⅲ proteins in urethral and anterior vaginal wall tissues of the mice in KO-VD and KO-VD+PES groups were significantly decreased（P<0.01）. Compared with WT-VD group， the expression levels of integrin β1 and calpain 2 proteins in urethral and anterior vaginal wall tissue of the mice in WT-VD+PES group were increased significantly （P<0.01）. Conclusion PES can improve the urodynamic function of the mice and promote the generation of pelvic floor collagen and promote the repairment of pelvic floor through the Cav 3.2 gene and its downstream integrin β 1 and calpain 2.

Objective To study the effects and mechanisms of Sanshen Ziwei Yin(SZY) on D cells and G cells of gastric mucosa in rats with chronic atrophic gastritis (CAG). Methods Rat models with CAG were established with synthesis method (active immunity,cold and cool gall and hot water irrigation). High and low-dose of SZY and Weimeisu (WMS,as control) were used for 50 days constantly. The pathological changes of gastric mucosa,morphological and number changes were observed before and after treatment. Results The number of G cells and D cells of rats with CAG decreased significantly (P<0.01), and degranulation presented in both two kinds of cells. The decrease of G/D ratio indicated that G cells descented more fast than D cells, that is to say, G cells were more sensitive to CAG. After treatment,the number of D cells increased obviously in treatment group, and apporached to normal levels at last,there was significant difference compared with WMS group. Conclusion SZY may have therapeutic effects on CAG through improving the inflammatory reaction of gastric mucosa to promote the regeneration of gland, endocrine and paracrine cells,and improving the regulation effects of gastrointestinal hormones on gastric function.

Objective To observe the effects of paclitaxel, mitomycin, 5-FU and cisplatin on the proliferation of tongue carcinoma cells and to guide the reasonable using of chemotherapeutic drugs and avoids the side-effects. Methods The fresh tongue carcinoma tissues from surgery were collected and unicellular suspension were prepared. Then paclitaxel, mitomycin, 5-FU and cisplatin were used to investigate the drug sensitivity on primary tongue carcinoma cells and a blank control group was set up. The inhibitory effects of chemotherapeutic drugs on the growth of tongue carcinoma cells and the cell cycle phases were observed by MTT colorimetry and flow cytometry. Results The inhibitory rates of paclitaxel, mitomycin and 5-FU on tongue carcinoma cell proliferation were 45.3%, 37.3%, and 36.2%, respectively, and in control group it was 2.1%, the difference was significant（P< 0.01）. But there was no significant different of inhibitory rate between cisplatin and control group. The results of flow cytometry showed that the amount of cells increased in G₀/G₁ phase, decreased in S phase and relatively increased in G₂/M phase after using paclitaxel, mitomycin and 5-FU compared with control group（P<0.01）. 　 Conclusion Paclitaxel, mitomycin and 5-FU can inhibit the growth of tongue carcinoma cells distinctively and should be considered prior to other drugs.

Objective To construct an eukaryotic expression plasmid containing the chimeric gene coding for the hemagglutinin-neuraminidase(HN) of newcastle disease virus(NDV) and human IL-18. Methods The enzymolysis and ligation were used to joint HN cDNA to IL-18 cDNA of pVAX1 IL-18 plasmid and at the same time the terminator of IL-18 cDNA was replaced, an eukaryotic expression vector of chimeric gene for HN and IL-18 was constructed. The recombinant plasmid was transfected into HeLa cells with liposome and the expression products were examined by hemagglutination test and Western blotting. Results The expression plasmid of pVAX1 IL-18 HN was contructed successfully confirmed by enzyme digestion identification. The expressions of IL-18 and HN chimeric genes were confirmed, and the expressed HN protein had higher hemagglutinative titer.Conclusion The constructed eukaryotic expression plasmid can express in vitro,and the expression products of the chimeric gene have good reactivity and specificity.

Objective To explore the effect of apoptosis on chondrocyte differentiation. Methods The apoptosis of costal chondrocytes of 5-week-old and 12-week-old rabbits was detected by methods of transmission electron microscopy and TUNEL. Results The proliferation and differentiation of chondrocytes was a gradual procedure. The apoptotic chondrocytes occurred at boundary from proliferation zone to hypertrophy zone by using transmission electron microscopy. The apoptosis existed in the whole cartilage zone, but the apoptotic percentage was highest in hypertrophy zone and increased with the age by using TUNEL method. Conclusion The proliferation and differentiation of costal chondrocytes from silence zone to bone zone of rabbits is a gradual procedure, in which apoptosis may play an important regulatory effect.

Objective To discuss the effect of CD147 on the glycolysis of the prostate cancer （PCa） LNCaP cells， and to provide new molecular markers and targets for the clinical diagnosis and treatment of PCa. Methods The lentivirus infection system was used to establish the cells silencing CD147 as LNCaP/shCD147 group， and the negative control group （LNCaP/scramble group） was set up at the same time. The expression levels of CD147 mRNA in the LNCaP cells in two groups were detected by real-time fluorescent quantitative PCR （RT-qPCR）method； the concentrations of lactic acid （LA）， pyruvate （PA），and ATP in the LNCaP cells in two groups were detected by the kits； the activities of pyruvate kinase （PK），6-phosphofructokinase 1 （PFK1） and hexokinase （HK） were detected by the kits，and the expression levels of HK2， phosphofructokinase of muscle（PFKM），and pyruvate kinase M2（PKM2） proteins in the LNCaP cells in two groups were detected by Western blotting method. Results Compared with LNCaP/scramble group， the expression levels of CD147 mRNA and protein in the cells in LNCaP/shCD147 group were significantly decreased（P<0.01）；indicating that the PCa LNCaP cell model with silencing CD147 was successfully constructed.Compared with LNCaP/scramble group， the concentrations of LA， PA， and ATP in the cells in LNCaP/shCD147 group were significantly decreased （P<0.05 or P<0.01）. Compared with LNCaP/scramble group， the activities of PK， PFK1， and HK enzymes in the cells in LNCaP/shCD147 group were decreased （P<0.05 or P<0.01）.The Western blotting results showed that compared with LNCaP/scramble group， the expression level of HK2 protein in the cells in LNCaP/shCD147 group was decreased （P<0.01）. Conclusion Silencing CD147 inhibits the glycolysis of LNCaP cells by regulating the glycolytic metabolites of glycolysis， rate-limiting enzyme activity， and related protein expression levels.

To explore the therapeutic effects of metallothionein (MT) on the H-22 liver neoplasm of BALB/c mice after administration with different concentrations of egg-milk with MT. Methods Seventy BALB/c mice were randomly divided into four groups:normal control (n=10)，tumor control (n=20)，low dose MT egg-milk group (n=20) and high dose MT egg-milk group (n=20).The mice excluding the normal control were injected with H-22 cells subcutaneously to build liver neoplasm models before they were administrated with egg-milk with MT for continuous two weeks.Lymphocyte proliferation ability and IL-2 activity in their spleen were measured with MTT method，the percentages of CD4⁺，CD8⁺ and CD25⁺ T cells were measured with flow cytometry，the killing activities of CTL and NK cells were detected with ³H-TdR method，the activities of IFN-γ and TNF-α were measured with cytopathic effect inhibition (CPEI) and L929 killed in vitro methods，respectively.The tumor volume of mice was detected at the 21th to 31th day. Results As compared with those in the tumor group，the tumor volume of the mice with H-22 cell hepatoma after administration with egg-milk with low dose and high dose MT decreased significantly at the 27th，29th and 31th day (P<0.05).The lymphocyte stimulating indexes in the low dose and high dose MT groups were significantly higher than that in the tumor control (P<0.01).Compared with the tumor control，the positive percentage of CD4⁺ T cells and the ratio of CD4⁺/CD8⁺ T cells increased significantly (P<0.01)，and the positive percentage of CD25⁺ T cells decreased significantly (P<0.05).The percentage of G₀/G₁ cells and the percentage of apoptotic cells in the high dose MT group were obviously lower than those in the tumor control (P<0.05)， and the percentage of G₂/M cells in the high dose MT group was obviously higher than that in the tumor control (P<0.05).The killing percentages of NK cells and CTL cells and the IL-2 and TNF-α activities in the high dose MT group were significantly higher than those in the tumor control (P<0.01 or P<0.05).While MT egg-milk had no effect on the IFN-γ activity. Conclusion Egg-milk with MT could build up the immune function of the mice with liver neoplasm and play a therapeutic effect on the tumor to some extent.

Objective To investigate genetic association between PCYT1A gene and schizophrenia in Chinese Han population from the Northeast area of China. Methods PCR-RFLP analysis was applied to　detect the genotype of PCYT1A gene in 115 family trios consisting of fathers, mothers and affected offspring with schizophrenia. The haplotype-based haplotype relative risk (HHRR) analysis and the transmission disequilibrium test (TDT) were used to process the genotype data. Results The genotypic frequency of the PCYT1A gene did not deviate from Hardy-Weinberg equilibrium in both the patient group （P>0.05） and the parent group （P>0.05）; The HHRR analysis did not show allelic association for the PCYT1A gene (χ²=0.011, P>0.05); the TDT analysis did not show preferential transmission of the two alleles (χ²=0.010, P>0.05). Conclusion There is no association between schizophrenia and the genetic polymorphism of rs3772108 locus of PCYT1A gene on chromosome 3.

Objective To study the effects of Panaxadiol Saponins (PDS) on glycemia, lipid and the improvement of insulin resistance (IR) in type 2 diabetes mellitus rats (T2DM). Methods Rats were randomly divided into normal group, model group, PDS 35,70 and 140 mg•kg^-1 groups and metformin group. Rats were fed by high fat diet for 10 d and injected with small dose of streptozotocin(STZ) 55 mg•kg^-1. Rats with plasma glucose over 11.1 mmol•L^-1we re assigned to T2DM model group,continually fed with PDS 35,70,140 mg•kg^-1•d-1 for 4 weeks and the values of fasting blood glucose (FBG) were assayed every week . Then FBG, Ins, C-peptide, FFA, TC, TG, HDL-c, LDL-c, MDA content and SOD activity were measured after 4 weeks. Results After treated with PDS for 4 weeks, the FBG in PDS 70, 140 mg•kg^-1 and metformin groups decreased significantly compared with normal group from the third week to the forth week (P<0.01); the liver glycogen　content was higher than that in model control (P<0.05 or P<0.01), the leve ls of TC, TG, FFA, TC/HDL-c, LDL-c/HDL-c and MDA were lower than those in model control (P<0.05 or P<0.01) . SOD activity, C-peptide content and ISI increased significantly (P<0.05 or P<0.01), while LDL-c, HDL-c changed little (P>0.05). Conclusion PDS could decrease the blood glucose, improve the disorder of the serum lipid and insulin sensitivity in T2DM rats.

Objective To investigate the relationship between the expressions of aquaporin 7 (AQP7) and aquaporin 8(AQP8) and the spermatogenesis of rat testis, and the effects of panaxadiol saponins(PDS) on expressions of AQP7 and AQP8. Methods Wistar rats of 4 and 12 weeks old were divided into saline control (Con group) and PDS groups. The rats in PDS group were injected with PDS in abdominal cavity (25 mg•kg^-1•d^-1，1 mL) every day, and saline was used in Con group with equal volume for 2 weeks. Semi-quantitative RT-PCR and Western blotting were employed to detect the mRNA and protein expression levels of AQP7 and AQP8. Results Expression levels of AQP7 and AQP8 were elevated when the age of rats increased; two weeks after injection of PDS, the expressions of AQP7 and AQP8 were upregulated in 4 weeks old rats, but the expressions of AQP7 and AQP8 in 12 weeks old rats were inhibited. Conclusion The expression levels of AQP7 and AQP8 are increased significantly with the sexual maturation; PDS can regulate the expressions of AQPs in testis.

Objective To discuss the effect of metformin on the hypertrophy of the H9C2 cardiomyocytes induced by isoproterenol （ISO）， and to clarify its possible molecular mechanism. Methods The H9C2 cells were cultured； 50 μmol·L^－1 ISO was used to establish the cardiomyocyte hypertrophy model，and they were used as ISO group；the cells without any treatment were used as control group. The expression levels of brain natriuretic peptide （BNP） in the H9C2 cardiomyocytes in two groups were detected by Western blotting method.The cells were divided into control group， ISO group， ISO+metformin group， and metformin group.The viabilities of the H9C2 cardiomyocytes in two groups were detected by CCK-8 method； the optimal drug intervention concentration and time of metformin were selected；crystal violet staining was used to observe the morphology and cross-sectional area of the H9C2 cardiomyocytes；real-time fluorescence qualitative PCR（RT-qPCR） method was used to detect the expression levels of atrial natriuretic peptide （ANP） in the cells in various groups；the morphology of mitochondriums in the H9C2 cardiomyocytes was observed by fluorescence probes； Western blotting method was used to detect the expression levels of optic atrophy 1 （OPA1） and mitofusin（MFN） proteins in the cells in various groups. Results Compared with control group，the cross-sectional area of the H9C2 cardiomyocytes in ISO group was increased significantly （P<0.05）， and the expression level of BNP protein was increased significantly （P<0.05）， suggesting that the cardiomyocyte hypertrophy model was successfully modeled. The RT-qPCR results showed that compared with control group， the expression level of ANP mRNA in the cells in ISO group was increased（P<0.05）； compared with ISO group， the expression levels of ANP mRNA in the cells in metformin+ISO and metformin groups were decreased（P<0.05）. The mitochondrium fluorescence probe observing results showed that the punctate mitochondrum in control group showed green fluorescence， mostly short rod-shaped， and a few mitochondrum were filamentously connected；compared with control group， the number of punctate mitochondrum in the H9C2 cells in ISO group was significantly increased，and the numbers of punctate mitochondrium in ISO+meformin and meformin groups were significantly decreased.The Western blotting results showed that compared with control group，the expression levels of OPA1 and MFN2 proteins in the cells in ISO group were decreased （P<0.05）；compared with ISO group， the expression levels of OPA1 and MFN2 proteins in the cells in ISO+metformin and metformin groups were increased（P<0.05）. Conclusion Metformin can alleviate the ISO-induced hypertrophy of the H9C2 cardiomyocytes，and its mechanism may be related to improving the mitochondrium fusion by up-regulating the expression levels of OPA1 and MFN2 proteins.

Objective To discuss the protective effectof follistatin-like 1 （FSTL1） on doxorubicin （DOX）-induced acute myocardial injury of the mice，and to clarify its related mechanism. Methods A total of 80 C57BL/6J mice were used to establish the acute myocardial injury models by intraperitoneal injection of DOX for one time.The mice were divided into 5 groups （n=8） according to the different doses （0，5，10，15， and 20 mg·kg^－1） of DOX，and didvided into 5 groups（n=8） according to the different intervention time（0，0.5，1.0，2.0，and 3.0 d）.The other 32 mice were randonly divided into normal saline group，FSTL1 group，DOX group， and DOX-FSTL1 group.The echocardiographic and hemodynamic indexes of the mice in various groups were detected；enzyme-linked immunosorbent assay（ELISA） method was used to detect the levels of tumor necrosis factor-α（TNF-α），N-terminal pro-brain natriuretic peptide（NT-proBNP） and cardiae troponin-T（cTn-T） in serum of the mice in various groups；the activities of superoxide dismutase （SOD）， the levels of malondialdehyde （MDA）， 4-hydroxynonenal （4-HNE） and 15-F_2t isoprostane in myocardium tissue of the mice in various groups were detected by oxidative stress kit；the expression levels of FSTL1 mRNA in myocardium tissue of the mice in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method； the expression levels of nuclear factor E2 related factor 2 （Nrf2） and FSTL1 proteins in myocardium tissue of the mice in various groups were detected by Western blotting method. Results Compared with normal saline group， the left ventricular ejection fraction（LVEF），left ventricular fraction shortening（ LVFS）， left ventricular systolic pressure（LVSP）， maximum rise rate of left ventricular pressure（+dP/dt_max），and maximum drop rate of left ventricular pressure（－dp /dt_max） of the mice in DOX group and DOX+FSTL1 group were significantly decreased（P<0.05）， and the heart rate（HR），left ventricular end-diastolic volume（LVEDV），and left ventricular diastolic pressure were increased （P<0.05）； compared with DOX group， the LVEF，+dP/dt_max， and －dp /dt_max of the mice in DOX+FSTL1 group were significantly increased（P<0.05），and the LVEDV was significantly decreased （P<0.05）. Compared with normal saline group， the serum levels of TNF-α， NT-proBNP and cTn-T in serum of the mice in DOX group and DOX+FSTL1 group were significantly increased （P<0.05）； compared with DOX group， the serum levels of NT-proBNP and cTn-T in serum of the mice in DOX+FSTL1 group were significantly decreased （P<0.05）. Compared with normal saline group， the activity of SOD in myocardium tissue of the mice in DOX group was significantly decreased （P<0.05）， and the levels of MDA， 4-HNE， and 15-F_2t-isoprostane were significantly increased （P<0.05），the level of 15-F_2t-isoprostane in serum of the mice in DOX+FSTL1 group was increased （P<0.05）； compared with DOX group， the SOD activity in myocardium tissue of the mice in DOX+FSTL1 group was increased （P<0.05）， and the levels of MDA， 4-HNE， and 15-F_2t-isoprostane were decreased （P<0.05）.Compared with 0 mg·kg^－1 DOX group， the levels of FSTL1 mRNA in myocardium tissue of the mice in the other groups were significantly decreased with the increasing of the doxorubicin dose （P<0.05）. Compared with DOX 0 d group， the levels of FSTL1 mRNA in myocardium tissue of the mice in the other groups were significantly decreased with the prolongation of the doxorubicin intervention time （P<0.05）. Compared with normal saline group，the expresison levels of FSTL1 and Nrf2 proteins in myocardium tissue of the mice in DOX group was decreased（P<0.05）；compared with DOX group， the expression levels of FSTL1 and Nrf2 proteins in myocardium tissue of the mice in DOX+FSTL1 group were increased （P<0.05）. Conclusion FSTL1 can alleviate the DOX-induced acute myocardial injury and improve the cardiac function by regulating the oxidative stress， and its mechanism may be reated to up-regulating the Nrf2 expression.

Objective To analyze the clinical and pathological characteristics of the mixed renal cell carcinoma （RCC） patient with co-existence of unilateral renal eosinophilic papillary renal cell carcinoma （OPRCC）and renal clear cell carcinoma （CCRCC）， and to improve the understandings of this disease. Methods The clinical data of one RCC patient with co-existence of OPRCC and CCRCC were retrospectively analyzed， and the diagnosis， treatment， and prognosis of the patient were analyzed in combination with the literature review. A 52-year-old male patient was admitted to the hospital due to a right renal mass detected by physical examination. The results of plain scan and enhancement CT of both kidneys showed space-occupying lesions of the right kidney， which was highly likely considered to be malignant. The laparoscopic partial nephrectomy was performed. Results The postoperative pathological results showed mixed RCC（lower pole of right kidney）.The results of immunohistochemical staining showed CK （AE/AE3） （partial +）， Vimentin （partial +）， EMA （partial +）， CK7 （+）， CD10 （+）， CAIX （partial +）， P504s （+）， PAX-8 （weak +）， TFE3 （－）， HMB45 （－）， MelanA （－）， SDHB （+）and Ki-67 （the positive rate was 2%）.The mixed RCC of the right kidney was diagnosed. The patient recovered quickly after operation and did not receive any adjuvant therapy. The CT examination results showed no local tumor recurrence and metastasis 3 months after operation， and no discomfort symptoms were found during the follow-up 6 months after operation. Conclusion The RCC patient with co-existence of OPRCC and CCRCC has no specific clinical manifestations， and the diagnosis mainly depends on the histopathological examination. Surgery is the first choice for the treatment；the malignant degree of the tumor is low， the progression is slow， the prognosis is good，and the long-term and close follow-up is still needed after operation.

Objective To investigate the promotion effect of Toll-like receptor 4（TLR4） adapters， myeloid differentiation factor 88（MyD88） and Toll/interleukin-1（IL-1） receptor（TIR） domain containing adaptor protein inducing interferon-β（TRIF） after gene knowout in M2 polarization of macrophages induced by lactate， and to elucidate the regulation mechanism of immune responses by lactate. Methods The murine macrophage cell line Raw264.7 were cultured in vitro， CRISPR/Cas9 gene editing system was constructed by lentivirus method， MyD88 and TRIF genes were knocked out（Myd88-KO group and TRIF-KO group），and the untransfection Raw264.7 cells were used as control group. The knockout effects of gene knowout were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. The experiment was divided into untreated Raw264.7 cell group， Raw264.7 + lactate group， MyD88-KO group， MyD88-KO + lactate group， TRIF-KO group and TRIF-KO+lactate group.The indexes were determined after 15 mmol·L^－1 lactate were induction for 24 h.The expression levels of M2 polarization phenotype molecules macrophage mannose receptor CD206 and arginase 1 （Arg1） mRNA in the cells in various groups were detected by RT-qPCR method， the morphology of cells in various groups was observed under light microscope， and the levels of tumor necrosis factor-α （TNF-α）， interferon-γ （INF-γ） and interleukin-10 （IL-10） in the cell supernatant in various groups were detected by enzyme linked immunosorbent assay（ELISA） method. The expression levels of nuclear factor-κB （NF-κB） signaling pathway-related proteins were detected by Western blotting method. The molecular docking simulation of lactate with TLR4， MyD88 and TRIF were carried out by Autodock software，and the binding was observed. Results After targeted gene was knocked out using CRISPR/Cas9 technique，compared with control group， the expression levels of MyD88 and TRIF mRNA and proteins in the cells in MyD88-KO group and TRIF-KO group were significantly reduced （P<0.05 or P<0.01）. After 24 h of lactate treatment， compared with untreated Raw264.7 cell group， the expression levels of markers of M2 polarization CD206 and Arg1 mRNA in the cells in Raw264.7+lactate group were significantly increased （P<0.05）； compared with MyD88-KO group， the expression levels of CD206 and Arg1 mRNA in the cells in MyD88-KO+lactate group were increased（P<0.05）； compared with TRIF-KO group， the expression levels of CD206 and Arg1 mRNA in the cells in TRIF-KO+lactate group were decreased （P<0.05）. The cells in Raw264.7+lactate group showed M2-polarized morphology， such as increased volume， obvious protruding pseudopods and increased proportion of conical cells； the cells in MyD88-KO+lactate group also showed M2-polarized morphological changes；the cells in TRIF-KO+lactate group showed no significant morphological changes. The ELISA results showed that compared with untreated Raw264.7 cell group，the TNF-α level in the cells in Raw264.7+lactate group was significantly decreased （P<0.05）， and the differences in INF-γ and IL-10 levels were not statistically significant （P>0.05）； compared with MyD88-KO group，the level of TNF-α in the cells in MyD88-KO+lactate group was significantly decreased （P<0.05）， the difference in INF-γ level was not statistically significant （P>0.05）， and the IL-10 level was significantly increased （P<0.05）；compared with TRIF-KO group，the TNF-α level in the cells in TRIF-KO+lactate group was significantly increased （P<0.05）， and the differences in the INF-γ and IL-10 levels were not statistically significant （P>0.05）.The Western blotting results showed that compared with untreated Raw264.7 cell group，the expression levels of TLR4 protein in the cells in Raw264.7+lactate group， MyD88-KO group， MyD88-KO+lactate group， TRIF-KO group and TRIF-KO+lactate group had no statistically significant differences（P>0.05）； the expression levels of nuclear factor-κB inhibitor protein-α（IκB-α） and p65 protein in the cells in Raw264.7+lactate group were significantly decreased （P<0.05）； compared with MyD88-KO group， the expression levels of IκB-α and p65 proteins in the cells in MyD88-KO+lactate group were decreased （P<0.05）； compared with TRIF-KO group， the expression levels of IκB-α and p65 proteins in the cells in TRIF-KO+lactate group had no statistically significant differences （P>0.05）.The number of hydrogen bonds formed by molecular docking of lactate with TLR4， Myd88 and TRIF was 2， 4 and 4， and the binding energies were －2.72， －3.24 and－3.66. Conclusion Lactate can inhibit the activity of IκB-α，and further inhibit NF-κB and thus promote M2 polarization through regulation of TRIF after treatment in the macrophages.

Objective： To investigate the clinicopathological features and prognosis-related factors of the patients with solitary fibrous tumor （SFT）， and to provide the evidence for its pathological diagnosis， clinical treatment and prognosis judgment. Methods The clinicopathological data of 86 patients with SFT who had undergone surgical resection from different systems were collected and divided into low （n=65）， medium （n=14） and high risk groups （n=7） according to the risk classification criteria. The general morphology of tumor was observed，HE staining and immunohistochemical staining were used to detect the morphology of SFT，and the follow-up data of the patients were obtained for the prognosis-related correlation analysis. Kaplan-Meier survival curve method was used to analyze the relationship between single clinicopathological factor in different parts and progression free survival of the patients， and Cox regression analysis was used to analyze the relationships between multiple factors and progression free survival of the patients. Results Of the 86 cases of SFT， 37 were male and 49 were female. There was no significant difference in the gender constituent ratio of the patients with SFT at different sites （P>0.05）. There was no significant difference in the age of patients with SFT at different sites （P>0.05）. There was no significant difference in the distribution of SFT at different sites among low，medium， and high risk groups （P>0.05）. There was significant statistical difference in the tumor diameter at different sites（P<0.01）. The microscope results showed that the shape and arrangement of tumor cells were diverse， and the spindle or oval cells were not arranged structurally in varying density； the characteristic antler like branching vessels and collagen fibers of varying thickness were common；most of the tumor cells were mild in shape and heterotypic， and the mitotic image was not obvious.The immunohistochemiscal staining results showed that the STAT-6 nucleus was diffusely and strongly positive； CD34， Bcl-2 and CD99 were positive in different degrees. A total of 61 cases were followed up for 2－139 months. Among them， 10 cases recurred， and the recurrence rates were 9% in low risk group， 14% in medium risk group， and 28% in high risk group， respectively. The univariate analysis results showed that there was a significant difference in the progression free survival between the patients with mitotic images<4/10 HPF and those with mitotic images ≥ 4/10 HPF （P<0.05）； there were significant differences in the progression free survival between high risk group and low，medium risk groups（P<0.05）. The multivariate analysis results showed that gender， age， tumor diameter and mitotic count were not the independent predictors of progression free survival of the patients （P>0.05）. Conclusion SFT can occur in many organs and systems of human body， and its morphology is diverse. The diameters of tumors in the central nervous system， upper respiratory tract and orbit are significantly smaller than those in female genital tract， abdominal cavity， subcutaneous soft tissue， lung and pleura. STAT-6 is a specific and sensitive index for SFT diagnosis； mitotic images ≥ 4/10 HPF and high risk classification are the risk factors for the progression free survival shortening； multivariate comprehensive analysis of tumor risk classification can not fully reflect the prognosis of the patients.

Abstract:Objective To  establish the animal model of arteriogenic erectile dysfunction(ED) and explore the pathogenesis of ED. Methods Thirty male Wistar rats were divided into two groups:experimental group (n=15),treated with ligating rat bilateral internal arteries;control group (n=15),treated with sham operation only. 4 weeks later,the erectile function of the rats was evaluated by injecting with apomorphine (APO) and  electrostimulation of cavernous nerves and recording of the　intracavernous pressure (ICP). The ultrastructures of the smooth muscle cells of cavernous of penis were observed with transmission electron microscope;the NOS level in the penis tissue was examined by NADPH diaphorase staining. Results In APO experiment,the penile erection times in experimental  group were decreased significantly compared with control group(P<0.01);the number of yawns was showed no obvious difference between  two groups（P>0.05）. In ICP experiment,the mean amplitude of ICP in experimental group was decreased significantly compared with control group (P<0.01). Electron microscopy showed that in the smooth muscle cells in experimental group,there were a significantly less number of mitochondria in the cytoplasm,less number of caveolae with fingerlike processes in the plasma membrane,increased number of vacuoles in the cytoplasm compared with control group. The NOS staining in experimental group was less intense compared with control group. Conclusion It is easy,practicable and reliable to establish the arteriogenic ED rat animal model by ligating rat bilateral internal arteries. Both the changes of ultrastructures and the low NOS level in penis tissue play roles in the occurrence of ED.

Objective To observe the protective effects of Fosinopril on myocardial ischemia-reperfusion injury in rats. Methods The myocardial ischemia-reperfusion model was induced by 30 min coronary occulusion and 24 h reperfusion in opened-chest anesthetized rats. The changes of aspartate aminotransferase(AST), lactate dehydrogenase(LDH), MB isoenzyme of creatine kinase(CK-MB), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and malondialdehyde (MDA) content in ser um were determined. Morphology of myocardial tissue was observed with optics and electron microscopes. Results Fosinopril decreased significantly the serum MDA content and AST, LDH and CK-MB activities as well as increased the serum SOD and GSH-Px activities. It also reduced edema and injury of ultrastructure in myocardial tissue. Conclusion Fosinopril has protective effects on myocardial ischemia-reperfusion injury, which may be related to scavenging the oxygen free radicals formed during reperfusion. Key words：Fosinopril/pharmacology; myocardial reperfusion inj ury; myocardial enzymes; free radicals; myocardium/pathology

Methods The pmiRFP670-N1 plasmid and pET-28a plasmid were doubly digested with restriction endonucleases EcoR Ⅰ and Not Ⅰ to construct the pET-miRFP670 prokaryotic expression vector. The LyP-1 DNA sequence was introduced through point mutation to construct the recombinant expression vector pET-miRFP670-LyP1； the recombinant expression vector with correct sequence was transformed into the E.coli BL21 cells and the prokaryotic expression amounts of fusion proteins induced under different temperatures （16 ℃ and 37 ℃） and different concentrations of isopropyl-β-D-thiogalactopyranoside （IPTG）（0.1，0.5，and 1.0 mmol·L^－1） were detected by SDS-PAG electrophoresis； the fusion protein was purified by Ni-NTA resin affinity， and the prokaryotic expression amount of the miRFP670-LyP1 protein was detected； the endocytosis morphology of the miRFP670-LyP1 fusion protein in breast cancer 4T1 cells was observed under fluorescence microscope. Results The double digestion of recombinant plasmid showed that two DNA bands of about 5 343 and 973 bp were obtained， which was consistent with the sizes of the pET-28a vector and miRFP670 gene fragment. The DNA sequencing results showed that the LyP1 sequence was successfully inserted into the pET-miRFP670 expression vector. The soluble protein expression amount of miRFP670-LyP1 fusion protein was higher at 16 ℃ than that at 37 ℃. The miRFP670-LyP1 fusion protein with high purity was obtained by purification with Ni-NTA resin. The fluorescence imaging results showed that the miRFP670-LyP1 fusion protein could be efficiently endocytosed by the breast cancer 4T1 cells. Conclusion The prokaryotic expression vector pET-miRFP670-LyP1 is successfully constructed，the soluble protein expression amount of the fusion protein is higher at low temperature （16 ℃） than that at normal temperature （37 ℃）， and the fusion protein with high purity is obtained by affinity chromatography，and the fusion protein is efficiently endocytosed by the breast cancer 4T1 cells and displays near-infrared fluorescence. Objective To construct the prokaryotic expression vector of tumor homing peptide（THPs）-near-infrared fluorescent protein（NIRFP）-miRFP670 LyP1 fusion protein， and to purified fusion protein， and to investigate the near infrared fluorescence characteristics of the fusion protein.

Objective To discuss the clinical and pathological characteristics of IgA nephropathy （IgAN） in the children and adults， and to clarify their clinical significances. Methods The clinical and pathological data of the patients diagnosed with primary IgAN were collected. According to their ages， the patients were divided into children group （n=160） and adult group （n=240）. The ages，genders，incidences of hypertension，time from onset to renal biopsy，estimated glomerular filtration rates，initial onset manifestations，clinical types，Lee’s grades， MEST-C scores，immunofluorescence types and ultrastructures of glomeruli of the patients in two groups were compared. Results The ratio of male to female in children group was 1.5∶1 and it was 1.1∶1 in adults group； compared with adult group， the incidence of hypertension and the time from onset to renal biopsy of the patients in child group were decreased （P<0.05）， while the eGFR was increased（P<0.05）. Compared with adult group， the percentages of gross hematuria and edema of the patients in child group were increased（P<0.01）， and the percentages of abnormal urine test， foam urine， and other manifestations were decreased（P<0.01）.Compared with adult group， the percentages of the patients with isolated hematuria and nephrotic syndrome in child group were increased （P<0.01）， while the percentages of the patients with isolated proteinuria and chronic nephritis were decreased （P<0.01）；compared with adult group， the percentage of the patients with grade Ⅱ in the Lee’s grades in child group was increased（P<0.01） and the percentage of the patients with grade Ⅴ in the Lee’s grades was decreased （P<0.01）.The light microscope obervation results showed that there were only focal mesangial cells and mesangial matrix hyperplasia in grade Ⅱ renal tissue of the patients in child group， rarely accompanied by glomerulosclerosis. The renal tubular and interstitial lesions were not significant； there were significant proliferations of the mesangial cells and matrix in grade Ⅴ renal tissue of the patients in adult group， with more glomerulosclerosis， and relatively severe renal tubular and interstitial lesions. Compared with adult group， the percentages of the patients with M1 and E1 in the MEST-C scores of the patients in child group were increased （P<0.05 or P<0.01）， and the percentages of the patients with S1 and T1/T2 were decreased （P<0.01）.The urinary protein grade of the patients in child group was positively correlated with M（r_s=0.462），E（r_s=0.342），and C（r_s=0.250）scores （P<0.01）；the urinary protein grade of the patients in adult group was positively correlated with M（r_s=0.217），E（r_s=0.145），S（r_s=0.187），T（r_s=0.269），and C（r_s=0.256）scores （P<0.01）；compared with adult group， the IgA+IgG+IgM deposition of the patients in child group was increased（P<0.05）， deposition rate of C3 was increased（P<0.01）， and the deposition rate of fibrinogen （Fib） was increased（P<0.01）. Conclusion There are significant differences in the clinical and pathological characteristics between the children and the adults with primary IgAN， which should be treated differently in the clinical diagnosis， treatment， and research.

Objective To observe the histological changes of nerve repaired by CO₂ laser welding and silk suture and investigate the advantages of laser welding. Methods Thirty Wistar rats were divided into 2 groups randomly. The sciatic nerves of right legs were cut in all rats, then in one group the nerves were repaired by laser welding, in the other group the nerves were repaired by silk suture as control. The histological changes of nerves were observed in both groups at 7, 14, and 21 days after operation. Results Compared with silk suture, laser welding had the advantages of fast healing and less inflammatory reaction. Conclusion Laser welding can improve the quality of nerve repairing, it is beneficial for the function recovery of nerve, therefore it is a ideal nerve repairing technique.

Objective To investigate the genetic association between the polymophism of progesterone receptor(PR) gene and susceptibility of uterine leiomyomas.Methods The method of PCR-RFLP was conducted to examine the genotypes of rs1145460 site of PR gene in 165 patients with uterine leiomyomas and 157 healthy women of Han descent. Hardy-Weinberg equilibrium for genotypic distribution was tested using the Chi-square (χ²) goodness-of-fit test.SPSS12.0 was applied to analyze the association between allelic gene and genotypic distribution and uterine leiomyomas.Results The genotypic frequency distribution of rs1145460 was not deviated from Hardy-Weinberg equilibrium in both patient group and control group (P>0.05).The genotypic（C/C,C/T and T/T）and allelic（C and T）frequencies of rs1145460 site had no significant difference between patient and control groups (P>0.05). Conclusion The polymorphism of rs1145460 site of PR gene may be not associated with the pathogenesis of uterine leiomyoma in Han women from north china.

Abstract:Objective To determine the plasma DNA level of patients with ovarian can cer and evaluate its potential clinical value.Methods Blood samples were collected from 30 ovarian cancer patients before and after surgery,and from 20 patients with ovarian benign tumor and 20 healthy women.The plasma DNA was extracted by commercial kit and detected by fluorescntmeter.Results The mean plasma DNA level in the cancer patients (25.33 μg•L^-1±17.69 μg•L^-1) was significantly higher than those in the benign tumor patients (10.28 g•L^-1±4.80 μg•L^-1) and normal control(7.60 μg•L^-1±3.87 μg•L^-1) (P<0.001).There was no significant difference between the benign and normal groups.The plasma DNA level of the patients in stage Ⅰ-Ⅱ (15.83 μg•L^-1±5.30 μg•L^-1) was significantly higher than that in the normal control(P<0.01) and was significantly lower than that in stage Ⅲ-Ⅳ(30.83 μg•L^-1±20.04 μg•L^-1) (p=0.005). The plasma DNA level of patients with metastasis(35.43 μg•L^-1±21.08 μg•L^-1) was significantly higher than that of patients with no metastasis(16.49 μg•L^-1±6.44 μg•L^-1) (P=0.006). The DNA level in the cancer patients one month after surgery (10.30±2.45 μg•L^-1) was much lower than before surgery(P<0.001),near that in the normal control.The overall survival of the patients with high plasma DNA level was much shorter than that of the patients with low DNA level(P=0.027).When the cut-off for diagnosis of ovarian cancer was 15.70 μg•L^-1,the sensitivity and specificity were 63.30% and 90.00%,respectively. Conclusion The plasma DNA level may have a potential use in early diagnosis and serve as a predict marker for metastasis potential and prognosis of ovarian cancer.

To investigate the regulatory effect of curcumin on proliferation inhibition and apoptosis in human prostate carcinoma PC-3M cells.Methods The PC-3M cells were divided into 4 treatment groups (10,20,30 and 40 μmol•L^-1 curcumin) and control group. The morphological alterations of PC-3M cells after treated for different time were observed by using inverse microscopy,the inhibition of cell proliferation was detected by MTT assay, the cell cycle phase was analyzed by flow cytometry,the apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining method,the protein level of caspase-3 in carcinoma cells was observed by SP immunohistochemistry. Results After being treated with curcumin, PC-3M cells grew round and small obviously and were against the wall under the inverse microscope. with the increasing of the concentration of cucurmin and the elongation of the treated time,the PC-3M cell growth was inhibited,the inhibition of proliferation of PC-3M cells increased in a time-and dose- dependent manner（P<0.01）;partial cells presented the morphological changes of apoptosis under the fluorescent microscope,the apoptotic rates of the control group and 4 treatment groups were（9.83±1.53）%,（19.50±1.00）%,（24.83±2.52）%,（30.17±2.08）% and（38.50±2.65）%, respectively;the cell number in S and G₂/M phase were increased, the cell number in G₀/G₁ phase was decreased;with the increasing of concentration of curcumin,the caspase-3 expression was increased in a dose- dependent manner（P<0.01）. Conclusion Curcumin could significantly inhibit the growth of PC-3M cells, the induction of apoptosis through up-regulating caspase-3 is probably one of its molecular mechanisms.

Objective To establish cell model of Parkinson’〖KG-3〗s disease (PD) and to approach the toxic effect of rotenone on dopaminergic neurons and its mechanism . Methods PC12 cells were differentiated into dopaminergic neurons and treated with various concentrations of rotenone. The morphological changes of PC12 cells were observed after treated with rotenone（0,10,25,50,75, and 100 nmol•L^-1）for 24,48 and 72 h , the cell viability was measured by MTT assay . Immunohistochemistry and immunofluorescence were used to observe the accumulation of α-synuclein in cytoplasm. AO/EB double staining was also adopted to test apoptosis. Results After differentiation PC12 cells shaped irregularly with big and long ecptomas and multiple cell conjunctions. After the treatment of rotenone cell ecptomas vanished gradually and cell bodies became smaller and smoother. The cell viability began to decline significantly at a concentration of 50 nmol•L^-1 for 24 h (P<0.05) ，and as concentration increased, the cell viability declined in a dose-dependent manner (P<0.01).As time prolonged, the cell viability behaved the same way. There was a significant difference between each time groups (P<0.01) .Immunohistochemistry demonstrated a brown round α-synuclein immunoactive mass in cytoplasm; cell immunofluorescence showed a strong greenα-synuclein immunoactive aggregation in cytoplasm. Viable apoptotic cells，non-viable apoptotic cells and necrosis cells were observed gradually as cell concentration increased and time prolonged. Conclusion Rotenone should be toxic to dopaminergic neurons，causing inclusion of α-synuclein aggregation and inducing apoptosis. The malmetabolism of α-synuclein and apoptosis induced by rotenone might play an important role in the pathogenisis of Parkinson’〖KG-3〗s disease .

Abstract:Objective To investigate the therapeutic effect of glycyrrhiza active substance on guinea pig model of vitiligo,and to clarify its mechanism on promoting synthesis of melanin,and to provide the basis for its clinical application.Methods Guinea pigs decolorized by H₂O₂ were used as experimental models,and normal control group was set up.After treatment of glycyrrhiza active substance in three doses groups(20,40,and 80mg·kg^-1),the therapeutic effect was observed by visual observation,macrography,histological examination as well as immunohistochemical study.Results The significant differences of skin color among control group,model group and treatment groups were observed by macrography.The total effective rates in 40 or 80 mg·kg^-1 groups were higher than that in control group (P<0.05 or P<0.01),and increased with the dose of glycyrrhiza active substance.Histological examination showed an evidently higher of melanin in basal cells,stratum spinosum and hair follicles in 40 or 80 mg·kg^-1 treatment groups compared with model group(P<0.01).Immunohistochemical results showed the expressions of HMB45 were higher in 40 or 80 mg·kg^-1 groups than that in model group (P<0.05 or P<0.01),and had dose-dependent relationship with glycyrrhiza active substance.Conclusion Glycyrrhiza active substance could increase the expression of HMB45 and promote synthesis of melanin,it shows certain efficacy in treatment of vitiligo,and it  can be used in treating vitiligo in clinic.

Objective To study the effects of formaldehyde on lung histomorphology and the level of lipid peroxide in mice after inhaled different amount of formaldehyde. Methods Forty male mice were randomly divided into 4 groups: one control group and three formaldehyde dose groups(20，40, and 80 mg•m-3). The mice were poisoned 2 hours one day for 5 weeks continuously, then were killed. Lung tissues of mice inhaled formaldehyde were observed under light microscopy (LM). The activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were detected biochemically in lung homogenate. Results It was manifestated as becoming bloodshots of pulmonary alveoli, expansion of pulmonary alveolus segment and inflammatory cells invasion under LM. The activities of SOD in 20,40, and 80 mg•m-3 dose groups were significantly lower than that in the control group, and the concentration of MDA was remarkably higher (P<0.01). The activity of SOD and the concentration of MDA had no remarkable differences between the different dose groups(P>0.05),but the former showed an increase activity in different dose groups, and the latter showed a reduced concentration in different dose groups. Conclusion Formaldehyde can injury free radical eliminated system to cause lung injury.

Objective To explore the linearic regularity of airways′ spasm and spasmolysis in hyperreactive guinea pigs. Methods Asthmatic guinea pig models were established with acetylcholine(Ach). Then the animals inhaled salbutamol. At 0, 15, 30 min and 1 h after inhalation of salbutamol, the animals were sacrificed, and the small and large airways were fixed with formaldehyde and embedded in paraffin wax to acquire the consecutive slides. The pathologic changes of both the large and small airway s were observed and the caliber changes of the airways were determined. Results The diameters of small and large airways of the control guinea pig models (0.41±0.10 and 1.78±0.41 mm,respectively) were significantly short than that in normal(0.55±0.20 and 2.41±0.50 mm, respectively)(P<0.05). Ach aerosol aspiration contracted the small and large airways similarly. After given salbutamol, the large airways began to contract at 15 min (1.99±0.55 mm) and lasted till 1 h (2.25±0.80 mm)（P<0.05）. However, no small airways dilated at 15 min or 30 min and began to dilate until 1 h(0.50±0.14 mm)（P<0.05）. Conclusion In asthmatic guinea pigs, the changes of spasmolysant in large airways and small airways are different, the large airways dilate first and small airways dilate later.

Abstract:Objective To establish the culture methods of submandibular gland cells from Wistar rats in vitro and lay a foundation for amplification of submandibular gland cells and gene therapy of salivary diseases.Methods The submandibular gland was obtained from Wistar rats which were one day old,after digested with LB3 and pancreatin，the submandibular gland cells were cultivated in DMEM/F12 culture fluid containing 2% fetal calf serum,epidermal gowth factor and insulin.The morphology of the culture and subculture cells was observed by phase contract microscope.The cell origin was identified by immunohistostaining.PAS staining was used to observe cell function of synthesis and secretion of polysaccharides.Results The morphous of primary cultured cells was round,triangle or polygon.At passage 2,the cells grew well,there was no change of morphous.The cells were positively stained with cytokeratin,E- cadherin and PAS,it indicated that the cells were epithelium origin and had the function of synthesis and secretion of polysaccharides.Conclusion The Wistar rat submandibular gland cells are successfully isolated and cultivated with enzyme digestion.The cultured submandibular gland cells at passage 2 still maintain their biocharacteristics.

Objective To discuss the protective effect of Yiyi Fuzisan on the myocardium ischemia and vascular endothelial dysfunction of the mice， and to clarify its possible mechanism. Methods Sixty male SPF grade C57BL/6J mice were randomly divided into blank group， sham operation group， acute myocardial ischemia （AMI） group， and low，medium and high doses of Yiyi Fuzisan groups， and there were 10 mice in each group；another forty C57BL/6J mice were randomly divided into normal saline group and low， medium， and high doses of Yiyi Fuzisan groups， and there were 10 mice in each group. The left anterior descending coronary artery （LAD） ligation method was used to establish the AMI mouse model， and corresponding drug intervention treatment was performed for 28 d. Western blotting method was used to detect the expression levels of endothelial nitric oxide synthase （eNOS） protein in myocardium tissue of the mice in various groups；arterial tension detection method was used to detect the tension of isolated aortic and relaxation rates of the mice in various groups；total nitric oxide（NO） detection kit was used to detect the levels of NO in serum of the mice in various groups；triphenyltetra zolium chloride（TTC） staining was used to observe the ischemia areas of myocardium tissue of the mice in various groups； HE staining was used to observe the pathomorphology of myocardium tissue of the mice in various groups；the postoperative survival rates of the mice in various groups were observed. The human coronary endothelial cells （HCAECs） were randomly divided into blank group， hypoxia group，hypoxia+ low dose of Yiyi Fuzisan group， hypoxia+medium dose of Yiyi Fuzisan group， and hypoxia+high dose of Yiyi Fuzisan group. Except for blank group， the cells in the other groups were cultured in the three gas incubator for 24 h to establish the hypoxia models. Griess test was used to detect the levels of NO in the HCAECs in various groups；fluorescence staining was used to detect the levels of mitochondrial membrane potential （MMP） and reactive oxygen species （ROS） in the HCAECs in various groups. Results Compared with before establishing the AMI model， the ST segment of electrocardiogram of the mice was significantly elevated after establishing the AMI model. The Western blotting results showed that compared with blank group， the expression levels of eNOS protein in myocardium tissue of the mice in sham operation group， AMI group，and low， medium， and high doses of Yiyi Fuzisan groups were decreased after injection of N-nitro-L-arginine-methylesterhydro chloride（L-NAME）（P<0.05）. Compared with saline group， the relaxation rates of thoracic aorta of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased （P<0.05）. Compared with blank group and sham operation group， the serum NO level of the mice in AMI group was decreased （P<0.05）， the serum NO level of the mice in high dose of Yiyi Fuzisan group was increased （P<0.05）； compared with AMI group， the serum NO levels of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased （P<0.05）. The TTC staining observation showed that compared with blank group and sham operation group， there were various degrees of myocardium ischemia of the mice in myocardium tissue of the mice in AMI group and low， medium， and high doses of Yiyi Fuzisan groups； compared with AMI group，the ischemia areas of myocardium tissue of the mice in low， medium， and high doses of Yiyi Fuzisan groups were decreased （P<0.05），and the percentages of ischemia areas of myocardium tissue were decreased.The HE staining results showed that the cardiomyocytes of the mice in blank group and sham operation group were neatly arranged， with clear and complete nuclei and uniform sizes，and no inflammatory cell infiltration was observed；the cardiomyocytes of the mice in AMI group showed significant myocardial tissue damage， with disordered arrangement，rupture and necrosis of cardiomyocytes， and infiltration of inflammatory cells；the myocardium tissue of the mice in low， medium， and high doses of Yiyi Fuzisan groups showed pathological recovery of myocardial tissue damage. On the 28th day after drug intervention， the survival rates of the mice in blank group and sham operation group were 100%； compared with AMI model group， the survival rates of the mice in low， medium， and high doses of Yiyi Fuzisan groups were increased（χ²=15.03，P=0.0102）.The Griess experiment results showed that compared with blank group， the levels of NO in the HEAEs in hypoxia group and hypoxia+low dose of Yiyi Fuzisan group were decreased （P<0.05）； compared with hypoxia group， the levels of NO in the HEAECs in hypoxia+medium dose of Yiyi Fuzisan and hypoxia+high dose of Yiyi Fuzisan groups were increased （P<0.05）. The fluorescence staining results showed that compared with blank group， the levels of MMP in the cells in hypoxia group， hypoxia group+low dose of Yiyi Fuzisan group， and hypoxia +medium dose of Yiyi Fuzisan group were decreased （P<0.05），while the levels of ROS were increased （P<0.05）； compared with hypoxia group， the levels of MMP in the HEAECs in hypoxia+low dose of Yiyi Fuzisan，hypoxia+medium dose of Yiyi Fuzisan，and hypoxia+high dose of Yiyi Fuzisan groups were increased （P<0.05）， while the levels of ROS in the HCAECs in hypoxia+low dose of Yiyi Fuzisan，hypoxia+medium dose of Yiyi Fuzisan，and hypoxia+high dose of Yiyi fuzisan groups were decreased （P<0.05）. Conclusion Yiyi Fuzisan can improve the pathological state of myocardium ischemia， restore the MMP， reduce the production of ROS， and improve the dysfunction of mitochondria and vascular endothelial cells by enhancing the biological activity of NO and increasing the vascular activity of aorta.

Objective To investigate the effects of Tripterium Regelii on urine protein and serum MMP-2, MMP9, and TGF-β₁ in nephritic rats. Methods The levels of serum MMP-2, MMP-9,and TGF-β₁ were detected by ELISA. Results The level of urine protein (mg/24 h) in nephritic rats treated with Tripterium Regelii was significantly decreased(pre-treatment: 327.38±48.67;post-treatment:170.38±32.62，P<0.01). The level of serum TGF-β₁ (ng•L^-1) was remarkably decreased too (model control: 287.60±43.37；model treatment:210.67±39.36，P<0.05)；the serum level of MMP-2 (μg•L^-1) was increased (model control: 240.63±38.34; model treatment:382.26±40.38，P<0.01)；the serum level of MMP-9 had a tendency to increase, but no significant difference was found (P>0.05). Conclusion Tripterium Regelii may have the effect of postponing glomerulosclerosis in rats.

Objective To investigate the protective effects of oral aprotinin on acute hepatic injury in rats. Methods Sixty Wistar rats were divided into 6 groups:normal control group (Ⅰ) , model control group (Ⅱ), low,middle, high doses of aprotinin groups (Ⅲ，Ⅵ，Ⅴ) and diammonium glycyrrhizinate control gro up (Ⅵ). The changes of activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), glutathione (GSH) and the liver weight coefficent were measured.The pathologic changes of liver tissue were measured. Results The serum ALT, AST, NO concentrations in rats and the liver weight coefficient in Group Ⅱ were increased significantly compared with Group Ⅰ (P<0.001, P<0.001, P<0.01, P<0.001), the serum GSH concentration in rats in Group Ⅱ was decreased significantly compared with Group Ⅰ (P<001). The serum ALT, AST, NO concentrations in rats in Group Ⅲ , Group Ⅳ and Group Ⅴ were decreased significantly compared with Group Ⅱ (P<0.001, P<0.001,P<0.05). The serum GSH concentrations in rats in Group Ⅲ , Group Ⅳ and Group Ⅴ were increased significantly compared with Group Ⅱ (P<0.01). The liver weight coefficients in Group Ⅲ , Group Ⅳ and Group Ⅴ were decreased significantly compared with Group Ⅱ(P<0.01,P<0.05,P<0.001). The order of pathologic change levels of liver tissue of acute hepatic injury was Group Ⅴ，Group Ⅳ，Group Ⅲ， Group Ⅱ. Conclusion The oral aprotinin has significantly protective fun ction on rats with acute liver injury.

Objective To investigate the effects of advanced glycation end products (AGEs) and Ginkgo biloba L. on proliferation, cell cycle and transforming growth factor β (TGF-β) expression of bovine retinal pericytes, and observe the protection of Ginkgo biloba L. on pericytes. Methods MTT colorimetric assay, flow cytometry, immunofluorescence were used for analysis. Results Ginkgo biloba L. could promote proliferation of pericytes undergoing AGEs. The Ginkgo biloba L. with concentration of 125~400 mg•L^-1 could promote proliferation of pericytes obviously (P<0.01) in a dose-dependent manner. Ginkgo biloba L. could inhibit the effects of AGEs on cell cycle of pericytes and induce apoptosis. Ginkgo biloba L. decreased the number of S-phase cells, increased G₂-M phase cells, decreased the number of apoptotic pericytes obviously compared with AGEs (P<0.01). Ginkgo biloba L. also down-regulated TGF-β protein expression induced by AGEs. Conclusion AGEs can damage bovine retinal pericytes through one of the pathways of oxidative stress, and antioxidant Ginkgo biloba L. can reduce the toxic effects of AGEs.

Abstract:ObjectiveTo assess the genetic affinities of the population from the Zhukaigou archaeological site in Inner Mongolia. MethodsThe mitochondrial DNA hypervariable region Ⅰ (HVRⅠ) in teeth of 9 individuals were amplified and sequenced. Different phylogenetic analyses were performed using mtDNA sequences among the ancient population，the Yinniugou ancient population and several modern Asian populations. The sequences were primarily assigned into each mtDNA haplogroups, and compared with modern populations. Results Fragments of mtDNA HVRⅠ, 364 bp in length, were recovered from 7 individuals from the Zhukaigou archaeological site. 7 sequence types and 15 polymorphism sites were found. Haplogroup distributions of the ancient individuals were investigated according to variations in the HVRⅠ sequence. ConclusionThe matrilineal genetic structure of the Zhukaigou site population is close to that of the later Yinniugou population and modern populations in this area. This aDNA study shows the continuity of the matrilineal genetic structure in the populations of the south central Inner Mongolia.

To study the assembly and release of adriamycin in vitro using magnetic thermosensitive liposomes treated with magnetic field and pyrogenation. Methods ll samples were divided into six groups : 21, 33, 39, 42, 45℃ and control groups according to the temperature of water bath. Lucifer yellow iodoacetamide (LY) was used as a fluorescence marker to adriamycin which was encapsulated by magntic thermosensitive liposomes. Liposomes was localized under the action of magnetic field, and adriamycin was released through the use of water baths with LY used as a fluorescence marker to observe the assembly and release. Temperature-treated liposomes were mixed with CT-26 cells to measure the binding of the relea sed LY-marked adriamycin to cell surface. Results he magnetic thermosensitive adriamycin liposomes was assembled directionally under magnetic field. The onset of LY release occurred near 33℃, and reached plateau above 42℃ when 90% of the LY was released. The binding with the HT-29 had positive correlation with the release of the LY-marked adriamycin. Conclusion agnetic field can make the mag netic thermosensitive liposomes drug to assemble directionally, hyperthermia can control the release of drug from magnetic thermosensitive liposomes.

Objective To investigate the effect of lncRNA-MIAT on M2-type polarization of tumor-associated macrophages， and to elucidate its possible mechanism. Methods The THP-1 cells were cultured in vitro. The lentivirus particles， which over expressed （LV-MIAT） and down regulated expression of lncRNA-MIAT（LV-shMIAT），and empty vector（LV-Vector） were transfected into the THP-1 cells. The THP-1 cells were activated into the macrophages， and the activated macrophages were co-cultured with the osteosarcoma （OS） MG63 cells by using Transwell co-culture system.The above co-culture systems were divided into MG63+LV-Vector group （positive control group），MG63+LV-shMIAT group， MG63+LV-MIAT group，and IL-4+LV-Vector group. The expression levels of lncRNA-MIAT in the THP-1 cells in various groups were detected；the percentages of M2-type macrophages in various groups were detected by flow cytometry； the levels of vascular endothelial growth factor （VEGF）， interleukin-10（IL-10） ，and transforming growth factor-β1（TGF-β1） in supernatant of the THP-1 cells in various groups were detected by ELISA method；the macrophages in each culture system were co-cultured with the human umbilical vein endothelial cell （HUVEC）； EDU staining was used to detect the proliferation activities of the HUVEC in various groups；the numbers of angiogenesis in the HUVEC in various groups were detected by tube formation assay；the expression levels of Janus kinase 1 （JAK1），signal transducer and activator of transcription 6 （STAT6），and phosphorylated STAT6 proteins in the THP1 cells and the expression levels of vascular endothelial growth factor receptor 2（VEGFR2），Notch1，and delta like protein 4（DLL4）in the HUVEC in various groups were detected by Western blotting method；the p-STAT6/STAT6 ratio was calculated.The OS tumor-bearing mouse model was constructed， and 36 nude mice were divided into LV-Vector group， LV-shMIAT group，and LV-MIAT group（n=12）.The volumes and weights of the tumor and the positive expression rates of CD163 and CD31 in tumor tissue of the mice in various groups were detected after interfering lncRNA-MIAT expression. Results The THP-1 cells with stable over-expression or down-regulation of lncRNA-MIAT were successfully established by lentivirus infection. Compared with LV-Vector group， the expression level of lncRNA MIAT in the cells in LV-shMIAT group was significantly decreased （P<0.05）， and the expression level of lncRNA MIAT in the cells in LV-MIAT group was significantly increased （P<0.05）. Compared with MG63+LV-Vector group， the levels of VEGF， IL-10，and TGF- β1 in the THP-1 cells in MG63+LV-shMIAT group were significantly decreased （P<0.05），and the expression levels of VEGFR2， Notch1， and DLL4 proteins in the HUVEC were significantly decreased （P<0.05），the levels of VEGF，IL-10，and TNF-β1 in the THP-1 cells in MG63+LV-MIAT and IL-4+LV-Vector groups were increased significantly （P<0.05 or P<0.01）；the expression levels of VEGFR2， Notch1，and DLL4 proteins in HUVEC were increased（P<0.05）， the percentage of M2-type macrophages，and p-STAT6/STAT6 ratio and expression level of JAK1 protein in the THP-1 cells were increased（P<0.05）；compared with MG63+sh-MIAT group， the VEGF level in the THP-1 cells in MG63+LV MIAT group was decreased significantly （P<0.05）， the levels of IL-10 and TGF-β were decreased significantly （P<0.05），the expression levels of VEGFR2， Notch1，and DLL4 proteins in the HUVEC were increased significantly （P<0.05）；compared with MG63+LV-Vector group， the proliferation activity and number of angiogenesis in the HUVEC in MG63+LV shMIAT group were significantly decreased（P<0.05），while the proliferation activities and number of angiogenesis in the HUVEC in MG63+LV-MIAT group and IL-4+LV Vector group were significantly increased （P<0.05）.The results of in vivo tumorigenesis experiment of the nude mice showed that compared with LV-Vector group， the volume and weight of transplanted tumor of the mice in LV-MIAT group were significantly increased （P<0.05），and the positive expression rates of CD163 and CD31 in tumor tissue were significantly increased （P<0.05）； the volume and mass of transplanted tumor and the positive expression rates of CD163 and CD31 in tumor tissue of the mice in LV-shMIAT group were significantly decreased （P<0.05）. Compared with LV-shMIAT group， the volume and weight of transplanted tumor of the mice in LV-MIAT group were significantly increased （P<0.05）， and the positive expression rates of CD163 and CD31 in tumor tissue were significantly increased （P<0.05）. Conclusion LncRNA MIAT may regulate the progression of OS by promoting the M2 polarization of macrophages and up-regulating the angiogenesis in tumor tissue.

Objective To investigate the correlations between the expression of　β-catenin and clinicopathologic parameters in bile duct carcinoma (BDC), and evaluate the potential prognostic value. Methods The expressions of β-catenin in 8 samples of normal bile ducts, 4 samples of congenital choledochal cyst and 25 samples of BDC were detected by immunohistochemistry staining　(S-P method). Results The β-catenin positive expression rate in BDC samples (100%) was obviously higher than those in samples of congenital choledochalcyst (75%) and normal bile ducts(62.5%) (Z=－3.03，P<0.01). The expressions of β-catenin in BDC were correlated with histological grade(Hc=9.35，P<0.01), infiltration grade (Hc=12.62，P<0.01)，perineural invasion (Hc=9.28，P<0.01), clinical stage(Hc=15.49，P<0.01), and were incorrelated with pathologic type(Hc=2.13，P>0.05), location(Hc=5.25，P>0.05), and severity of jaundice (Hc=6.06，P>0.05). Conclusion β-catenin is correlated with some clinicopathologic parameters of BDC and can be used as a valuable prognostic marker of BDC.

Objective To study the regulatory effects of polysaccharide Fb from Huangmo (Fb) on immunological function in mice. Methods 40 healthy adult mice were randomly divided into control group (C), CTX immune-suppressed group (CTX), Fb treated group (Fb), Fb combined with CTX treated group (M). The influences of Fb in NK cell activity were determined by the method of NK mediated cytotoxicity test；the multiplicative capacity of lymphocyte was assayed by lymphocyte transformation rate (LTT) of T cell，and the mice spleen T lymphoblast multiplication analytical method was used to measure the IL-2 level. The expressions of MMPs in related immune system were measured by gelatin zymogram electrophoresis. Results Fb increased the NK cell activity, LTT and captivity of IL-2 in immune-suppressed mice（P<0.05），and also promoted the LTT in normal mice （P<0.01）.The weights of spleen and thymus in Fb and M groups were increased, and the body weights of mice in Fb and M groups were increased remarkably than that in CTX group（P<0.01）. The activities of MMP-2（P<0.05）and MMP-9（P>0.05）indicated a decreased tendency in CTX group, the activity of MMP-9 were increased significantly in Fb group and M group（P<0.01）. Conclusion Fb has an activating effect on immune system and may protect and promote the proliferating and repairing of immune system in immune-suppressed mice. Fb can promote the LTT of T cell significantly and may up-regulate some other immune indexes slightly.

Objective To study the effects of growth hormone (GH) on morphology of cardiac muscle and skeletal muscle and hormone levels in Wistar rats. Methods The GH was given with subcutaneous injection for 15 days, the level of serum GH was determined by radiation-immune method; the body weight and the ratio of organ weight to body weight were determined; the cell appearances of cardiac muscle and skeletal muscle were observed under microscope. The control group was set up. Results The level of serum GH and rat body weight in experimental group were obviously higher than that in the control group, but the ratio of organ weight to body weight was not obviously different in two groups; musculature hypertrophy and cell nucleolus increasing were observed under microscope, there were no capillary vessel hyperplasia and inflammatory soakage. Conclusion GH can induce hypertrophy of cardiac muscle cells and skeletal muscle cells but not interstitial proliferation.

To detect the expression of TrkA in the brain of the Wistar rats during the embryonic phase. MethodsAdult Wistar rats were fed in one cage.It was E0 day when sepermia were found in vaginal suppository and vaginal smear.The expressions of TrkA in the brain of Wistar rats during various brain regions were observed by immunohistochemistry on E13,E15,E17,E19 and E21 day. Results On E13 day no obvious expression of TrkA was observed in the embryonic brain. The expression of TrkA had an obviously increasing tendency in the cortex of telencephalon from E15 day to E17 day. And on E17 day, the positive rate of the expression of TrkA reached the second peak. While on E19 day, the expression of TrkA had a temporary decreasing. Then on E21 day the expression of TrkA reached the highest peak. The striatum and hippocampus developed later than the telencephalon till E17 day, and the expressions of TrkA during these stages were same as the cerebrum. Conclusion From E15 to E17 day, obvious expressions of TrkA are observed in the embryonic telencephalon, and it accords with embryonic induction.

Objective To compare the microvessel density (MVD) and the expressions of vascular endothelial growth factor (VEGF), CD44V⁶ and CD34 in ductal breast carcinoma between primary tumor and lymph node metastasis for researching the mechanism of metastasis and the corresponding reaction of body. Methods The expressions of CD34, VEGF, and CD44V⁶ were detected and analyzed in 30 cases of breast cancer with lymph node involvement (LN⁺) and 8 cases of breast cancer without lymph node involvement (LN^-) by immunohistochemical technique. Results Higher MVD was observed in the primary lesions than in the metastatic foci (P<0.05), and the expressions of VEGF were not significantly different between these two positions. There was a positive correlation between the expressions of VEGF and MVD in the primary tumor, and there was not a correlation between the expressions of VEGF and MVD in the metastatic foci in the lymph nodes. The expressions of CD44V⁶ were not significantly different between primary lesions and metastatic foci. The expressions of VEGF, CD34 and CD44V⁶ in the LN⁺ primary lesions were higher than those in the LN^- lesions (P<0.05). Conclusion The ability of angiogenesis between primary lesions of ductal breast carcinoma and its corresponding metastatic foci in the lymph nodes are different. Some elements in the metastatic foci can suppress the ability of angiogenesis.

Abstract:Objective To study the influence of anti-sense oligonucleotides(ASODN) of  Survivin on carcinoma cell proliferation by the technique of ASODN.Methods  Survivin-ASODN was synthesized according to Survivin gene sequence.The SMMC-7721 cells were transfected with fluorescence labled Survivin-ASODN.At the same time,blank control,lipofectamine control and SODN control groups were set up.The Survivin gene expression levels were measured by PCR.The proliferation inhibitory rates of Survivin-ASODN transfected cells were analyzed with MTT method.Results The transfection rate reached 60% of fluorescence labled Survivin-ASODN in SMMC-7721 cells.After transfected with ASODN at 100,200,300,400 and 600 nmol?L-1,the Survivin gene expression levels were decreased.[JP2]The inhibitory rates were (3.87±1.67)%，(9.21±2.21)%，(15.21±3.18)%，(21.32±[JP]3.43)%，and (32.62±3.74)%,respectively,at 24 h after transfection. In 300 nmol?L-1 ASODN group,the inhibitory rates were (29.21±4.12)% at 48 h and (44.21±3.32)% at 72 h.Compared with blank control group,lipofectamine  group and SODN group,the inhibitory rates in ASODN group showed significant difference (P＜0.05).Conclusion Survivin-ASODN has inhibitory effect on cell proliferation in dose-dependent and time-dependent manner.

Objective To study the effects of Modified Papanicolaou stain,Wright-Giemisa stain and Coomassie brilliant blue stain on the rate of morphogically normal sperm and the rate of acrosome intactness. ethods Semen smears from 55 male infertility patients were stained according to Wright-Giemisa, Modified Papanicolaou and Coomassie brilliant blue methods. The rate of morphologically normal sperm and the rate of acrosome intactness were evaluated. Results ①No significant differences were found in the percentage of morphologically normal sperm between the three staining methods (P>0.05). Comparing with Modified Papanicolaou stain and Coomassie brilliant blue stain, the rate of Neck/Midpiece defect sperm ,as well as tail defect sperm , significantly decreased after Wright-Giemisa stain（P<0.05）. ②No significant difference was observed in the rate of sperm with type I acrosome and the rate of sperm with type Ⅳ acrosome between the three staining methods (P>0.05). However, the rate of sperm with type Ⅱ or Ⅲ acrosme was higher using Modified Papanicolaou stain than using the other two stains（P<0.05）. ③The significantly positive correlationship was found in the rate of normal morphologically sperm (r=0.535 P<0.01，r=0.601,P<0.01 , r=0.329,P<0.05), as well as in the rate of acrosome intactness （r=0.604, P<0.01，r=0.682,P<0.01, r=0.446,P<0.01）,among the three staining methods. Conclusion The three staining methods stain would be reliable and applicable to detect the percentage of human morphologically normal sperm and the rate of acrosome intactness. Among the three staining methods, Modified Papanicolaou stain would be better to be used in examing the percentage of human morphologically normal sperm, and Wright-Giemisa stain would be more suitable for detecting the rate of acrosome intactness.

Objective Using a recombinant adenoviral vector carrying the human BMP-2 gene (Ad-BMP-2) to transfect rabbit bone marrow stromal cells (BMSC) to study the effects of Ad-BMP-2 transfection on cell differentiation. Methods The rabbit BMSC cultured in vitro were transfected by Ad-BMP-2. Seven days later, the expressions of BMP-2 and collagen type Ⅱ in these cells were determined by in situ hybridization and immunohistochemical analysis, and the alkaline phosphatase (AKP) activity of the BMSC was observed. Results There were positive results in cytoplasm in experimental group: the brown-yellow hybridization signals were observed and that indicated the expressions of BMP-2 and collagen type Ⅱ of Ad-BMP-2 infected BMSC by in situ hybridization, the yellow pigmenting were observed by immunohistochemical analysis but not in nuclei, the brown-black particles were seen by AKP staining. But in control group, these were negative. Conclusion Ad-BMP-2 could infect BMSC with high efficiency and promote the conversion to osteoblast and chondrocyte.

Objective To determine the progesterone receptor (PR), estrogen receptor (ER), and Bcl-2 protein expressions in endometrial carcinoma and their clinical significances. Methods The expressions of PR, ER, and Bcl-2 in a total of 104 frozen tissues of endometrial carcinoma and 30 cases of paracarcinoma tissues were examined by immunohistochemistry. Results ①The expressions rates of PR, ER, and Bcal-2 protein in endometrial carcinoma were 65.38%, 46.15%, and 50.84%, respectively. They were 66.66%, 60.00%,and 26.66%, respectively, in parcarcinoma tissues. Statistically difference was found in the expressions of Bcl-2 between two groups (P<0.05), while it was not found in the expressions of ER and PR. ②Expression of PR was positively correlated with that of Bcl-2 (r=0.216，P<0.05). There was no relation between ER and Bcl-2. ③Positive expression rate of ER in 16 lymph node metastasis cases was 50.00%, those of PR and Bcl-2 were 12.50% and 25.00%,respectively. Positive expression rates of ER, PR, and Bcl-2 in 88 non lymph node metastasis cases were 45.45%, 75.00%,and 59.09%,respectively. Expressions of PR and Bcl-2 had statistically differences in the cases of lymph node metastasis or non-metastasis　(P<0.05), while ER had not.　④Expression of PR was higher in highly differentiated endometrial adenocarcinoma than that in poorly differentiated one. Conclusion Positive expression rate of Bcl-2 in endometrial carcinoma is high, it is positively correlated with PR expression. PR and Bcl-2 positive expression rates in lymph node metastases cases are low. PR and Bcl-2 might be available prognostic factors.

Abstract:Objective To investigate the relationship between two excision repair cross-complementation group 1(ERCC1) polymorphisms, 8092C>A and 19007G>A, and susceptibility to acute lymphoblastic leukemia (ALL) in children . Methods A case-control study consisting of 183 children ALL patients and 190 controls in a Chinese population was performed to examine the ERCC1 genotype. Results For the ERCC1 8092C> A polymorphism, individuals carrying the CC genotype had a significant high risk when compared with carrying one A allele gene (AA /AC) patients (P = 0.030). For the ERCC1 19007G >A polymorphism, no association was found between ERCC1 polymorphism and ALL risk. Stratification for sex with regard the ERCC1 8092C > A and 19007G >A polymorphism form showed a highly significant risk (OR=1.94, 95% CI 1.09-3.41, P=0.020;OR=2.36, 95%CI 1.05-5.27, P=0.037) of ALL among males whereas that of females did not show any significant risk. For the ERCC1 8092C> A polymorphism, individuals under 10 years old carrying CC genotype had significantly risk (OR=1.76，95% CI 1.04-2.99; P=0.04). However, the ERCC1 19007G >A polymorphism was not significantly associated with ALL in age. Conclusion These results suggest that the ERCC1 8092CC gene polymorphism is related to the occurrence of ALL in Chinese children.

To explore the effect of lead acetate on the growth of murine mesenchymal stem cells in vitro. Methods 40.00,60.00 and 100.00 μmol•L^-1 of lead acetate were used in the culture of colony-forming unit-fibroblast(CFU-F),the effect on the rate of colony-forming and the rule of variation were observed. Results The rates of colony-forming were(3.30±0.20), (2.40±0.10) and(1.57±0.21)/10⁵ ,when the doses of lead acetate were 40.00 ,60.00 and 100.00 μmol•L^-1 and there were significant differences compared with control group(4.20±0.20)/10⁵, P<0.05). The rates of colony-forming had significant differences between lead acetate with different doses groups (P<0.05). Dose-effect relationship was analyzed with linear correlation and correlation coefficient（r=-0.960,P<0.01）. Colony cells of control group presented fusiform shape while cells of 40.0 μmol•L^-1 of lead acetate group presented irregularity or round shape cultivated in vitro after 6 d. Conclusion The lead acetate suppresses the rate of colony-forming of CFU-F significantly in vitro and the inhibitory effect increases with the increasing of doses of lead acetate.

Objective To construct an eukaryotic expression vector of human ERβ (hERβ) full length gene which named pEGFP-C1-hERβ and transfect it into hormone-independent prostate cancer cell line PC-3M. Methods The complete gene of hERβ（1 593 bp） was obtained with the technique of RT-PCR by using human ovary tissue mRNA as template which obstained from the ovarian cyst patients. The PCR product was connected with pMD18-T vector and sequenced automatically，and then the connected product and the expression vector pEGFP-C1 were digested by restrictive enzyme Hind Ⅲ and BamH Ⅰ， respectively，and the pEGFP-C1-hERβ vector was constructed by using gene recombinant technique. The plasmid was detected by endonuclease digestion and PCR，and then was transfected to PC-3M cells by lipid reagent. Results The clone of the complete hERβ gene and the construction of expression vector were finished successfully by endonuclease digestion and sequenced automatically. The product of the endomuclease digestion was as long as the human complete hERβ gene (1 612 bp)，and sequence analysis suggested that the hERβ sequence detected by PCR was identical to that published in GenBank (NM_001437). The expression of green fluorescence protein (GFP) was observed under the fluorescence microscope. Conclusion The eukaryotic expression vector pEGFP-C1-hERβ is constructed and it expresses in PC-3M cells successfully.

Abstract:Objective To study the inhibitory effect of hammerhead ribozyme targeting connective tissue growth factor (CTGF) on collagen I synthesis and cell cycle progression of human hepatic stellate cell line (LX-2) cells.Methods Hammerhead ribozyme cDNA targeting CTGF mRNA plus two self-cleaving sequences were inserted into pTriEx2 vector to construct a recombinant vector pTriCTGF-Rz.LX-2 cells were transfected with either pTriEx2 or pTriCTGF-Rz and further stimulated with or without TGF-1. There were five groups in the experiment:control group,pTriEx2 group,pTriCTGF-Rz group,pTriEx2 plus TGF-β1 group,and pTrCTGF-Rz plus TGF-β1 group.Semi-quantitative RT-PCR was used to detect the levels of CTGF mRNA and collagen Ⅰ mRNA.ELISA and flow cytometry were used to detect the levels of collagen Ⅰ secretion and cell cycle.Results Transfection of pTriCTGF-Rz into LX-2 cells reduced the CTGF mRNA and collagen Ⅰ mRNA levels as well as collagen 　Ⅰ protein level compared with pTriEx2 group(P<0.05).TGF- 1-induced CTGF mRNA and collagen Ⅰ mRNA transcription were decreased in LX-2 cells transfected with pTriCTGF-Rz compared with control groups (P<0.05).Furthermore,in the LX-2 cells transfected with pTriCTGF-Rz,the proportion of cells at G₀-G₁ phase increased (P<0.05)　 and that of cells at S phase decreased(P<0.01) compared with control group.Conclusion The hammerhead ribozyme targeting CTGF has inhibitory effects on human hepatic stellate cell-mediated fibrosis and it may become another candidate for gene therapy in the future.

Objective Abstract：Objective To explore the effects of arsernic trioxide (As₂O₃) on telomerase activity and cellular apoptosis of human colon cancer cell line CCL-187.Methods CCL-187 cells were divided into control group，1.0 μmol•L^-1As₂O₃ group and 2.5 μmol•L^-1As₂O₃ group，all groups were cultivated for 1 to 6 d.The inhitory rate of cell growth was detected by MTT assay.The morphologic changes of CCL-187 cells were observed under electron microscope.The expression of hTERT at mRNA level was analyzed by RT-PCR.Telomerase activity was determined by TRAP-PCR-ELISA.Results The differentiation of cells was found in 1.0　μmol•L^-1 As₂O₃ group，but apoptosis in 2.5　μmol•L^-1 As₂O₃ group，and at the same time ，the telomerase activity was reduced and the hTERT-mRNA expression was downregulate in 1.0 μmol•L^-1 As₂O₃ group and 2.5 μmol•L^-1 As₂O₃ group in a time-dependent manner.There were significant differences compated with control group（P<0.01 or P<0.01）;there were also significant differences between 2.5 μmol•L^-1 As₂O₃ group and 1.0 μmol•L^-1 As₂O₃ group（P<0.01）.The inhibitory rate of cell growth was closely negatively correlated with the downregulation of hTERT-mRNA（1.0 μmol•L^-1 As₂O₃ group,r=－0.803，P<0.01；2.5 μmol•L^-1 As₂O₃ group,r=－0.697，P<0.01） and the reduction of telomerase activity（1.0 μmol•L^-1 As₂O₃ group,r=－0.836， P<0.01；2.5 μmol•L^-1As₂O₃ group,r=－0.792，P<0.01）.The reduction of telomerase activity was closely correlated with the downregulation of the hTERT-mRNA（1.0 μmol•L^-1 As₂O₃ group,r=0.956，P<0.01； 2.5 μmol•L^-1 As₂O₃ group,r=0.988，P<0.01）.Conclusion As₂O₃ can induce differentiation and apoptosis of CCL-187 cells.Its mechnism may be ralated to reducting the telomerase activity through downregulation of the hTERT-mRNA expression.

Objective To investigates the influences of focal cerebral ischemia on CRH，NGF and HPA in hypothalamus of rats. Methods Thirty-two senile Wistar rats including fourteen female and fourteen male rats were divided into seven groups at random and there were four rats per group. One group was control group, the other six groups were operation groups. Operation groups consisted of ischemia 15 min group and ischemia 15 min groups in which rats were reperfused for 1，6 h and 1，2，4，9 d. The expressions of CRH and NGF in hypothalamus of focal cerebral ischemia senile rats were observed dynamically with the immunohistochemical method. Results There was a little expressions of CRH and NGF in hypothalamus in control group, the expressions of CRH and NGF in hypothalamus didn′t change during 1 h after ischemia reperfusion in operation groups, then the expressions of CRH and NGF in hypothalamus disappeared transiently at 6 h after ischemia reperfusion, but after that the expressions of CRH and NGF in hypothalamus increased obviously at 2 d after ischemia reperfusion. Conclusion The activity of CRH secretory cells in hypothalamus is not immediately in the strength phase after cerebral ischemia; the activity strength effect of CRH secretory cells in hypothalamus exists in the later stage after cerebral ischemia; NGF is possibly important to the activity of HPA.

Objective To observe the effect of enzymes-hydrolyzed wheat bran (EHWB) on antioxidant system in diabetic rats.Methods Diabetes mellitus models were induced by alloxan inject ion and EHWB was used to test its effect on anti-oxidative capacity; the EHWB was given intragastrically (i.g.), sodium ferulate (SF) as a positive group. The level of blood glucose, total anti-oxidative capacity (T-AOC), the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and xanthine oxidase (XOD), and the content of malondiadehyde (MDA) were detected respectivel y in serum and liver of rats. Results EHWB significantly decreased the level of blood glucose and increased the level of T-AOC, as well as the activities of GSH-Px and SOD , and decreased the activity of XOD and the content of MDA. However, there was no difference between the groups feeding SF and EHWB. Conclusion EHWB can be used to decrease blood glucose and improve antioxidant ability in diabetes mellitus rats，the effect of EHWB is similar to SF. The effect of decreasing blood glucose of EHWB is related to protecting liver.

Objective To study the clinical efficacy of differential diagnosis between malignant and benign solitary pulmonary nodules (SPNs) with single-location dynamic enhanced computed tomography (CT). Methods Thirty-nine patients were studied with single-location dynamic enhanced CT. Patterns of time-density curve were drawn. Peak height and increment of CT value were calculated. 〖WTFZ〗Results Malignant and benign SPNs showed quite different patterns in time-density curve. Enhanced degree of pulmonary carcinoma ［(41.9±2.8）Hu］was higher than that of the tuberculosis［（11.7±7.85)Hu］, which had significant difference (P<0.01). However, here was no significant difference of enhanced degree between pulmonary inflammatory［（43.6±7.7)Hu］ and pulmonary carcinoma SPNs (P>0.10).But the time-density curve of malignant SPNs tend to achieve rapid enhancement- to-peak attenuation and eventually reached a plateau. Inflammatory SPNs showed gradual enhancement without obvious attenuation. The curve of tuberculosis was placid. Conclusion Single-location dynamic contrast enhanced CT is a non-invasive option and can provide quantitative information about blood flow patterns of SPNs.

Objective To study the protective effects of Sibelium on neuron injury in rats and its mechanism. Methods The damage model of cultured cortical neuron induced by H₂O₂ was established. The malondialdehyde (MDA) was detected by TBA method, and the breaking rate of DNA was determined by diphenylamine method. The change of transudatory rate of lactate dehydrogenase (LDH), the effects of Sibelium on those indexes described above and its interaction with •OH in vitro were observed. Results The intoxication of H₂O₂ was decreased by Sibelium. Sibelium singnificantly reduced the transudatory rate of LDH, the breaking rate of DNA. The content of MDA in culture and the clearance rate of •OH was increased with the increasing of the dosage of Sibelium. Conclusion Sibelium can obviously protect the cultured cortical neuron against intoxincation of H₂O₂ in rats.

To explore the methods for isolating and culturing dendritic cells derived from rat bone marrow in vitro，and provide reference for immunological research and clinical application. Methods The bone marrow mononuclear cells(BMMNC) were obtained from bone marrow of Fischer344 rat’s limbs through density gradient centrifugation;and the mature DC was gained by BMMNC cultured in medium with recombinant rat granulocyte-macrophage colony stimulating factor (rrGM-CSF),recombinant rat interleukin-4 (rrIL-4) and recombinant rat tumor necrosis factor -α(rrTNF-α).In group 1,the rat’[KG-*3]s ovarian cancer cell line NuTu-19 freeze-melt antigens were added on the third day and the tumor cells lysates pulsed DC was acquired;in group 2,as control,nothing was added.Then the shape of DC was observed,the cell-surface phenotype and the proliferation rate were detected.ResultsThe mature DC could be induced by BMMNC co-culturing with relative cytokines,and the expressions of cell-surface phenotypes such as OX62,CD86 and MHC-Ⅱ in group 1 were higther than those in control group (P<0.01).When the ratios of stimulating cells(SC) to reaction cells(RC) were 1∶3,1∶10,1∶30  and 1∶100,the stimulating indexes(SI) in group 1 were higher than those in control group(P<0.01). Conclusion The culture method of DC from bone marrow of Fischer344 rats is established,which means the mature DC can be acquired from BMMNC.

Objective To study the effects of Gross Saponins of Tribulus Terrestris (GSTT) on thrombosis in vivo and in vitro. Methods ①The effects of GSTT on thrombus wet weight and thrombosis time of rats in vivo and the thrombus length, thrombus wet and dry weight in rats in vitro were observed by using the methods of neck artery-vein side-way thrombosis and artery thrombosis induced by electricity stimulation.②The effects of GSTT on the blood viscosity in blood stasis rats were observed. Results ①GSTT with doses of 20.8 and 31.2 mg•kg^-1 could significantly decrease thrombus wet weight (P<0.05 or P<0.01), and prolong thrombosis time (P<0.01), and shorten the thrombus length in vitro(P<0.05), and decrease thrombus wet and dry weight in vitro (P<0.01 or P<0.05). ②GSTT with doses of 20.8 and 31.2 mg•kg^-1 can significantly decrease blood viscosity (P<0.01). Conclusion GSTT has the effects of antithrombosis, and its mechanism is related to lowering blood viscosity and improving blood hemorheology.

Objective To analyze the effect of Shenqi Dihuang Decoction in the treatment of membranous nephropathy （MN） based on network pharmacology method and molecular docking technique，and to clarify its mechanism. Methods The effective components and corresponding target proteins of 7 herbs of Shenqi Dihuang Decoction were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP）；the target genes of MN were collected from the GeneCards database，OMIM database， and Drugbank database； the network of “components-compounds-targes” of Shenqi Dihuang Decoction was constructed by Cytoscape software； the protein-protein interaction network was constructed through STRING database and Cytoscape software， and the topological analysis was performed to identify the core targets； the molecular docking for the core targets with the active ingredients acting on them was performed to screen the core components of Shenqi Dihuang Decoction for the treatment of MN.Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Gene and Genomes （KEGG） pathway enrichment analysis were performed by Bioconductor R software. Results A total of 55 effective components，214 drug action targets，3 655 MN targets， and 144 intersection targets were obtained，including 5 core targets，which were protein kinase B1 （AKT1）， epidermal growth factor （EGF）， vascular endothelial growth factor A （VEGFA）， tumor protein P53 （TP53）， and epidermal growth factor receptor （EGFR）； three potential core components were screened based on the molecular docking technology， including quercetin， luteolin， and diosgenin；the GO enrichment analysis results revealed 2 268 biological processes， 53 cell compositions，and 175 molecular functions； the KEGG pathway enrichment analysis results revealed 169 related signal pathways. Conclusion Shenqi Dihuang Decoction may play a role in treatment of MN by reducing the level of inflammation， inhibiting the renal fibrosis and maintaining the integrity of glomerular filtration barrier.

Objective To investigate the pregnancy outcomes of fresh embryo transfer in the same infertile patients receiving the GnRH antagonist （GnRH-ant） program and the GnRH agonist （GnRH-a） program for ovulation， and to provide basis for the selection of ovulation induction program. Methods Using retrospectively research method， the clinical data and laboratory examination data of 74 patients （148 cycles in total） received GnRH-a program and GnRH-ant program treatment for ovulation within 2 years were collected. The effects of ovulation promotion， embryo development and clinical outcome of fresh cycle transplantation were analyzed. Results The ovulation induction detection results showed that compared with GnRH-a group， the total dose and time of gonadotropin （Gn） use， and endometrial thickness of the patients in GnRH-ant group were decreased （P<0.05 or P<0.01）； the level of luteinizing hormone （LH） on the human chorionic gonadotropin （HCG） day was significantly increased （P<0.01）；there were no significant differences in the number of oocytes retrieved and the levels of progesterone（P） and estradiol（E2） on the HCG day （P>0.05）. The embryo status detection results showed that compared with GnRH-a group，the 2 pronucleus（2PN） rate， abnormal fertilization rate，2PN cleavage rate， available blastocyst formation rate and high quality blastocyst formation rate in GnRH-ant group were increased， but there were no significant differences （P>0.05），while the good embryo rate and available embryo rate in GnRH-ant group were significantly increased （P<0.05）.In clinical pregnancy outcome， there were no significant differences in the number of embryos transferred， ectopic pregnancy rate， abortion rate， implantation rate and clinical pregnancy rate in two groups （P>0.05）. Conclusion GnRH-ant program could be a more suitable plan for ovulation due to the higher good embryo rate， avaliable embryo rate， lower Gn dose and shorter Gn using time compared with GnRH-a program.

To study the regulation of the huangmo alkali-dissolved polysaccharide (Fb) on expression of MMPs and its anti-hepatocarcinama effect in tumor-bearing mice. Methods Transplanted liver cancer models of mice were set up,the mice were equally divided into control group, CTX group, and three treatment groups of low,middle,high doses of Fb (Fb₂₀，Fb₄₀ and Fb₈₀). The inhibitory rate (IR) was determined to observe the anti-tumor effect in vivo; Gelatin zymography and quantified analytical system were used to detect gray-scale value of straps to observe the expressions of MMP-2 and MMP-9; the regulation of Fb on the protein expressions of MMP-2，9 and TIMP-1 was analyzed,the positive rate was detected by immunohistochemistry. Results IR of Fb₂₀,Fb₄₀ and Fb₈₀ groups (38.7%， 51.2% and 58.3%) were higher than that in control group （P<0.05 or P<0.01）; the expression rates of TIMP-1 in Fb₄₀ and Fb₈₀ groups (70.0% and 84.5%) were higher than those in control group and CTX group(41.7% and 44.4%)(P<0.05), the expressions of MMP-9 were down-regulated in Fb₄₀（47.7%）and Fb₈₀　(44.4%) 　 groups，but there was no difference,compared with control group (60.0%) (P>0.05).The expression of MMP-2 in Fb₄₀ group（20.0%）was lower than that in control group（58.3%）（P<0.05）. Conclusion Fb has strong activity to restrain the tumor growth and metastasis in transplanted liver cancer H₂₂，the anti-tumor mechanism of Fb probably correlates with its effect on regulating the activities of protein expressions of MMPs and TIMPs.

Objective To explore the regulative mechanism of apoptosis-associated gene proteins on the adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays. Methods Kunming male mice were irradiated with the inductive doses (D1: 25, 50, 75, 100 and 200 mGy; dose rate: 12.5 mGy•min^-1) and the challenging dose (D2: 1.5 Gy; dose rate: 287 mGy•min^-1). The time interval between D1 and D2 was 6 h. The expressive levels of thymocyte apoptosis-associated gene proteins were measured with flow cytometry. Results As compared with the sham-irradiation, the positive percentage of thymocyte Bcl-2 protein expression decreased significantly in D2 group (P<0.05), Bax increased significantly (P<0.05), and Bcl-2/Bax decreased significantly (P<0.001); p53 increased significantly (P<0.001). As compared with D2 group, the positive percentage of thymocyte Bcl-2 protein expression increased in varying degree in D1+D2 group of 25~75 mGy D1, Bax decreased in varying degree, and Bcl-2/Bax increased significantly (P<0.01); p53 decreased significantly (P<0.001 or P<0.05). Conclusion The apoptotic thymocytes in the adaptive response of thymocyte apoptosis in mice induced by irradiation with 25~75 mGy decrease significantly due to the increase of apoptosis-associated gene Bcl-2 protein expression and Bcl-2/Bax, the decrease of Bax and p53 protein expressions.

Objective To explore the mechanism of proconditioning to hypoxia. Methods The changes of NOS neuron of mouse brain in proconditioning to hypoxia were detected by histochemistry method. Results Compared with control mice, the number, colour, shape, dendrites and axons number and length of NOS positive neurons in cerebral cortex was not changed after mice were exposed to hypoxia for one time, but the dendrites and axons of NOS neuron became thicker. Compared with control mice, NOS neurons in cerebral cortex was not changed after mice were exposed to hypoxia for four times. There were little NOS neurons in hippocampus of control mice and NOS neurons were light positive. Compared with control mice, the number of NOS positive neurons in hippocampus increased and NOS neuron became strong positive after mice were exposed to hypoxia for one time. Compared with mice exposed to hypoxia for one time, the number of NOS positive neurons in hippocampus decreased and their colour became light after mice were exposed to hypoxia for four times. Conclusion The changes of NOS neurons can enhance generation of proconditioning to hypoxia.

Objective To investigate the preliminary effect of oxymatrine　(OMT) on inflammation and its mechanism. Methods Fifty mice were randomly divided into 5 groups: normal saline group, dexamethasone group and OMT groups with different doses (70,140,and 210 mg·kg^-1). The extent of ear swelling was observed in inflammation model caused by dimethyl benzene in mice. Forty-eight Wistar rats were randomly divided into 6 groups:normal saline group, dexamethasone group and OMT groups with different doses (50，100,and 150 mg·kg^-1),L-NAME+OMT 100 mg·kg^-1 group. Paw edema volume induced by dextran were observed in rats. Results The extents of ear swelling were decreased in dose-dependent manner after intraperitoneal injection of OMT 140 and 210 mg·kg^-1 in mice compared with control (P<0.05 or P<0.01). Paw edema volumes were reduced after intraperitoneal injection of OMT 100 and 150 mg·kg^-1 in rats compared with control (P<0.05). L-NAME enhanced the effect of anti-inflammation of OMT. Conclusion OMT can reduce the extent of ear swelling induced by dimethylbenzene in mice and the paw edema volume caused by dextran in rats.Its anti-inflammatory effect may be not fully mediated by NO-cGMP pathway.

Objective To clone the sequence of the cDNA of mouse endostatin (mEndostatin) coding area and construct an expression vector containing Egr-1 promoter, IFNγ and endostatin genes. Methods Full length mEndostatin was obtained with the technique of RT-PCR and using mouse hepatocyte mRNA as template, pMD18T- mEndostatin was sequenced automatically and an expression vector containing Egr-1 promoter, IFNγ, and endostatin genes was constructed with gene recombinant technique. Results It was proved that the cloned mEndostatin cDNA to be completely identical with that reported in the literature and the recombinant plasmid containing Egr-1 promoter, IFNγ, and endostatin genes was constructed successfully. Conclusion mEndostatin cDNA was cloned and the expression vector pEgr-IFNγ-mEndostatin was constructed successfully.

Abstract:Objective To explore the relation of the expressions of MDM2 and VEGF in osteosarcoma with the pathological parameters and prognosis of the tumor.Methods The expressions of MDM2 and VEGF were detected with immunohistochemical(SP) method in specimens from 56 cases of osteosarcoma.The correlation between the expressions of MDM2,VEGF and pathological grade,metastasis and prognosis was analyzed statistically.while 8 cases of fibrous dysplasia of bone were used as negative control group.Results The positive rates of MDM2 and VEGF in osteosarcoma were 64.3%(36/56) and 67.9%(38/56)， respectively.MDM2 and VEGF didn’[KG-*3]t express in negative control group.The expression of MDM2 and VEGF were not significantly correlated to the pathological grades of the osteosarcoma,but which were significantly correlated with tumor metastasis and prognosis (P<0.05).The five-year survial rate in patients with positive expression of MDM2 and (or) positive expression of VEGF was significantly lower than that in negative control group(P<0.05).The patients with positive reaction for both MDM2 and VEGF had the lowest survival rate, but the patients with negative reaction for both MDM2 and VEGF had the highest survival rate,the difference was significant (P<0.01).Conclusion There is an intimate correlation between the overexpression of MDM2 and VEGF in osteosarcoma and its unfavorable prognosis.MDM2 cooperating with VEGF probably plays a role in angiogenesis of osteosarcoma and is an useful parameter to evaluate prognosis of osteosarcoma.

To investigate the contribution of transplantation of ex vivo expanded endothelial outgrowth cells （EOCs）from peripheral blood on repair of balloon injured vessels in rabbits. Methods Animals were divided into two groups(n=8): EOCs transplantation group and control group. The mononuclear cells（MNCs）were isolated from rabbit peripheral blood by density-gradient centrifugation and cultivated in EGM-2, yielding EOCs. A rabbit model of balloon catotid injury was used to evaluate the effects of EOCs transplantation on endothelial regeneration and inhibition of neointimal formation. Immediately after denudation, EOCs（the second passage）in 100 μL saline or 100 μL saline alone (control) were administered into the lumen of injured artery. Meanwhile the cells were labeled by CM-DiI for cells tracking. Four weeks after transplantation, rabbits were killed. The endothelial regeneration rate (RA/TA) and IA/MA ratio in fluorescence-labeled EOCs were detected. Results Four weeks after transplantation, fluorescence-labeled cells were found in the media, neointima and on the luminal surface of injured segments. Evans blue staining result demonstrated that RA/TA ratio in EOCs transplantation group (89.1%±6.3％) was much higher than that in control group (62.1%±7.5%, P<0.01). Meanwhile, compared with control group, there was a significant decrease of neointimal formation in EOCs transplantation group. The IA/MA ratios were 0.88±0.14 and 0.44±0.06, respectively(P<0.01）. Conclusion Transplantation of ex vivo expanded EOCs to balloon-injured arteries can accelerate reendothelialization significantly and reduce neointimal formation.

Objective To assess the effects of tissue inhibitor of metalloproteinase-3 (TIMP-3) on proliferation and invasion of PC-3M cell line in vitro. Methods The adhesion assay was used to detect the effects of TIMP-3 on cancer cell adhesion abilities. MTT colorimetry was used to detect the proliferation abilities of PC-3M cells. Monolayer and Boyden chamber invasion assay were performed to assess cell invasion abilities. Results The adhesion rate and proliferation rate of transfected cells containing TIMP-3 gene were significantly lower than those of cells transfected with empty plasmid and untransfected cells (P<0.05). The cell invasion number and index of the former were significantly lower than the others (P<0.05). The differences of adhesion rate, proliferation rate and invasion index between transfected empty plasmid cells and untransfected cells weren′t statistically significant (P>0.05). Conclusion TIMP-3 can significantly inhibit the proliferation and invasion of human prostate carcinoma PC-3M cell line.

Objective To investigate the effect of paired box transcription factor 3 （PAX3） gene silencing on the differentiation of the P19 cells into the cardiomyocyte-like cells，and to clarify its possible mechanism. Methods Dimethylene maple （DMSO） was used to induce the differentiation of the P19 cells into cardiomyocyte-like cells， and the morphology of the cells during the induction process was observed. The cells were collected on the 0th day （0 d group） and 10th day （10 d group） after induction and differentiation.The P19 cells were infected with sh-PAX3 Lentivirus.The cells were divided into control group，sh-NC group（negative control group）and sh-PAX3 （PAX3 interference）.Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to verify the positive expression of cTnI in P19 cells silenced with PAX3 gene. The P19 cells infected with lentivirus were treated with Notch signal agonist Jagged1 and were induced and differentiated with DMSO for 10 d. The cells were divided into DMSO group， DMSO+sh-NC group， DMSO+sh-PAX3 group， DMSO+Jagged1 group and DMSO+sh-PAX3+Jagged1 group. RT-qPCR method was used to detect the expression levels of GATA binding protein 4 （GATA4）， atrial natriuretic peptide （ANP）， cardiac troponin （cTn）T，PAX3，Notch1， Notch intracellular domain （NICD）， and Hes family bHLH transcription factor 1 （Hes1） mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of PAX3，Notch1， NICD， and Hes1 proteins in the cells in various groups； immunofluorescence assay was used to detect the positive expression of cTnI in the cells the differentiation rates of the cardiomyocyte-like cells in various groups. Results On the 10th day， after induction and differentiation，the cluster of cardiomyocyte-like cells with spontaneous rhythmic contraction could be observed.The RT-qPCR and Western blotting results showed that after lentivirus infection， compared with control group and sh-NC group， the expression levels of PAX3 mRNA and protein in the P19 cells in sh-PAX3 group were decreased （P<0.05）， indicating that the P19 cells silenced with PAX3 gene were successfully constructed. The RT-qPCR results showed that compared with 0 d group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in 10 d group were increased （P<0.05）， while the expression levels of PAX3，Notch1， NICD， and Hes1 mRNA were increased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+sh-PAX3 group were decreased （P<0.05）， while the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+Jagged1 group were increased （P<0.05）； compared with DMSO+sh-PAX3 group， the expression levels of GATA4， ANP， and cTnT mRNA in the cells in DMSO+sh-PAX3+Jagged1 group were increased （P<0.05）. The Western blotting results showed that compared with 0 d group， the expression levels of PAX3， Notch1， NICD， and Hes1 proteins in the cells in 10 d group were increased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+sh-PAX3 group were decreased （P<0.05），while the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+Jagged1 group were increased （P<0.05）； compared with DMSO+sh-PAX3 group， the expression levels of Notch1， NICD， and Hes1 proteins in the cells in DMSO+sh-PAX3+Jagged1 group were increased （P<0.05）.The immunofluorescence results showed that after lentivirus infection， the positive expression of cTnI protein was found in the cells in various groups. Compared with DMSO group and DMSO+sh-NC group， the positive expression of cTnI protein in the cells in DMSO+sh-PAX3 group was decreased， and the differentiation rate of the cardiomyocyte-like cells was decreased （P<0.05）； compared with DMSO group and DMSO+sh-NC group， the positive expression of cTnI protein in the cells in DMSO+Jagged1 group was increased，and the differentiation rate of the cardiomyocyte-like cells was increased （P<0.05）； compared with DMSO+sh-PAX3 group， the positive expression of cTnI protein in the cells in DMSO+sh-PAX3+Jagged1 group was increased，and the differentiation rate of the cardiomyocyte-like cells was increased （P<0.05）. Conclusion PAX3 gene silencing may inhibit the differentiation of the P19 cells into the cardiomyocyte-like，and its mechanism may be realized by decreasing the PAX3 gene expression and regulating the Notch signaling pathway.

Objective To analyze the changes of cholesterol levels of the patients with acute coronary syndrome （ACS）， and to explore the predictive value of residual cholesterol （RC） levels in the occurrence of ACS. Methods A total of 220 ACS patients were selected as ACS group； 220 healthy people underwent physical examination at the same time were selected as control group. The general informations including serum levels of total cholesterol （TC）， low density lipoprotein cholesterol （LDL-c） and high density lipoprotein cholesterol （HDL-c） of the subjects in two groups were collected， and the levels of serum non-high density lipoprotein cholesterol （non HDL-c） and serum residual cholesterol （RC） were calculated. Logistic regression analysis was used to analyze the risk factors of the ACS patients and their correlations with the levels of various lipid components. The receiver operating characteristic curve（ROC） and the area under （AUC） were used to evalue the predictive values of various blood lipid components in the ACS. Results The single factor analysis results showed that compared with control group，the percentages of the patients with hypertension and smoking in ACS group were increased（P<0.01），the levels of TC， RC， LDL-c，non HDL-c of the patients in ACS group were increased（P<0.01），and the level of HDL-c was decreased（P<0.01）.The multivariate Logistic regression analysis results showed that hypertension （OR=0.496， P=0.005）， smoking （OR=0.458， P=0.002），RC level （OR=24.051，P<0.01）， and non HDL-c level（OR=1.711， P<0.01） were the independent risk factors for the occurrence of ACS. The ROC curve analysis results show that compared with non-HDL-c level， the AUC of RC level was larger，which had the higher predictive value for the occurrence of the ACS patients. Conclusion RC level is an independent risk factor for the occurrence of ACS， and has certain predictive value for the occurrence of ACS.

Abstract:Objective To study the role of NF-kappaB( NF-κB) on islet cell apoptosis in high concentration glucose in vitro.Methods Pancreatic islet cells wereisolated from Kunming mice and these cells were cultivated in medium with different concerntrations of glucose: G1 (5.6 mmol•L^-1),G2(7.8 mmol•L^-1),G3(11.1 mmol•L^-1),G4(16.7 mmol•L^-1),G5(22.2 mmol•L^-1),G6(27.6 mmol•L^-1).After the slet cells were cultivated for 72 h,insulin secretion was evaluated by radio-immunity technique.The NF-κB expression was detected by immunocytochemistry and cytochrome C(Cyt.C) release was detected by immunofluorescence technique.Apoptosis of islet cells was determined by Hoechst33342 assay. Results Insulin secretion was enhanced and NF-κB activity was increased（P<0.05）when the concentrations of glucose were 5.6-11.1 mmol•L^-1,but Cyt.C release and apoptosis didn’t change. When the concentration of glucose exceeded 16.7 mmol•L^-1,the apoptotic percentages of islet cells were increased signficantly accompanying with the glucose concentration elevation compared with G₁,G₂ and G₃ groups(P<0.05),the NF-κB activity and Cyt.C relesae were significantly increased （P<0.05）.Conclusion High glucose can induce the apoptosis of islet cells and decrease insulin secretion,enhance the activity of NF-κB and Cyt C release.It suggests that the activation of NF-κB may be related to apoptosis of islet cells,and the inhibition of NF-κB may protect islet cells.

Methods The GES-1 cells from normal gastric mucosa，gastric cancer MGC-803 cells，MKN-45 cells，and HGC-27 cells were collected. The hsa_ circ_ 0009735 small interfering RNA and negative control were transfected into the MGC-803 cells， and the gastric cancer MGC-803 cells were divided into si-hsa_ circ_ 0009735 group and negative control group；the control plasmid and hsa_circ_0009735 over-expression plasmid were transfected into the HGC-27 cells， and the gastric cancer HGC-27 cells were divided into OE-hsa_circ_0009735 group and control plasmid group， and the MGC-803 cells transfected with autophagic plasmid pcDNA-eGFP-LC3 were divided into blank group and different concentrations （0.25， 0.50， 1.00 and 2.00 mg·L^－1） of rapamycin groups.The expression levels of hsa _ circ_ 0009735 in the gastric cancer tissue and cells were detected by real-time fluorescence quantitative PCR （RT-qPCR）method；laser confocal microscope was used to observe the morphology of the MGC-803 cells and the number of autophagosomes in the MGC-803 cells in various groups； Transwell chamber experiment was used to detect the number of migration cells in various groups； flow cytometry was used to detect the percentages of the cells at different cell cycles in various groups； Western blotting method was used to detect the expression levels of microtuble-associated protein light chain 3Ⅱ（LC3 Ⅱ），microtuble-associated protein light chain 3Ⅰ（LC3 Ⅰ），E-cadherin， Cyclin D1， Vimentin，and N-cadherin proteins in the cells in various groups. Results The PCR products were sequenced and showed cyclization sites，which was matched with the sequences of circ RNA hsa_circ_0009735. Compared with the sequences of adjacent tissue， the expression level of hsa_ circ_ 0009735 in gastric cancer tissue was increased （P<0.05）； compared with GES-1 cells， the expression level of hsa_ circ_ 0009735 in the MGC-803 cells was increased （P<0.05）.There was no significant change of the morphology of the MGC-803 cells among 0.25 and 0.50 mg·L^－1 rapamycin groups.Compared with blank group，the number of granules in the MGC-803 cells in 1.00 mg·L^－1 rapamycin group was increased and the boundary was unclear； in 2.00 mg·L^－1 rapamycin group， more MGC-803 cells died. The laser confocal microscope observation results showed that the number of autophagosomes in the MGC-803 cells in 1.00 mg·L^－1rapamycin group had no significant difference compared with 2.00 mg·L^－1 rapamycin group （P>0.05）.Compared with blank group， the ratio of LC3 Ⅱ/LC3 Ⅰ in the MGC-803 cells in different concentrations of rapamycin groups were increased （P<0.05）， the expression levels of hsa_circ_0009735 were decreased （P<0.01）. Compared with negative control group， the expression level of hsa_circ_0009735 in the MGC-803 cells in si hsa_ circ_ 0009735 group was decreased（P<0.01）， the ratio of LC3 Ⅱ/LC3 Ⅰ was increased （P<0.01），the number of migration cells was decreased （P<0.01）， the percentage of cells at G₁ phase was increased （P<0.05）， the percentage of cells at S phase was decreased （P<0.01）， the percentage of cells at G₂ phase was decreased （P<0.05）， and the expression levels of Cyclin D1， Vimentin，and N-cadherin proteins in the cells were decreased （P<0.05）； compared with control plasmid group， the expression level of hsa _ circ -0009735 in the HGC-27 cells in OE-hsa_ circ_0009735 group was increased significantly （P<0.01）， the ratio of LC3 Ⅱ/LC3 Ⅰ was decreased （P<0.01）， the number of migration cells was decreased （P<0.01）， the percentage of the cells at G₁ phase was decreased （P<0.01）， the percentage of the cells at S phase was increased （P<0.05）， the percentage of cells at G₂ phase was increased （P<0.05）， the expression level of E-cadherin protein in the cells was decreased （P<0.05）， and the expression levels of Cyclin D1， Vimentin，and N-cadherin proteins in the cells were increased （P<0.05）. Conclusion High expression of hsa_circ_0009735 in the GC cells may promote the EMT process and affect the migration and the cell cycle， and inhibit the autophagy. Objective To investigate the expressions of circular RNA hsa_circ_0009735 in the gastric cancer tissue and cells， and to explore its effects on the epithelial mesenchymal transformation（EMT）， migration， cell cycle，and autophagy of the gastric cancer cells.

To investigative the changes in renal function and histopathological in caulis aristolochiac manshuriensis(CAM)-induced injury in rats. MethodsMale Wistar rats were randomly divided into control group(n=10) and administration group(n=30).After CAM were administered by gavage to rats for 3,7,15 d,the blood and 24 h-urine samples were collected,24 h-urinary protein excretion,urinary retinol-binding protein (RBP),serum creatinine (Scr) and blood urea nitrgen (BUN) were examined.The pathological changes in kidney were observed at various time and compared with control group.Results① At various time after administration of CAM,the Scr,BUN and 24 h-urinary protein excretion levels were not increased compared with control group.Compared with control group,after administration of CAM for 3 d,urinary RBP content was significantly increased( P<0.01),and the values increased with the elongation of time.② The degeneration and necrosis in tubules were major pathological changes in rats after administration of CAM for 3 d，and the pathological changes were aggravation as treatment time increased. Conclusion The degeneration and necrosis in tubules are major pathological changes in rats after administration of CAM.

Objective To discuss the regulatory effect of long chain non-coding RNA （lncRNA） metastasis associated transcript 1 （MALAT1） on the activation of the hepatic stellate cells （HSC） during the progression of liver fibrosis， and to clarify its mechanism. Methods The serum samples were collected from 25 healthy volunteers （n=25，healthy group） and 25 liver fibrosis patients［ mild liver fibrosis group （n=12）and severe liver fibrosis group （n=13）］. The mouse HSC were divided into control group，transforming growth factor-β1（TGF-β1） group， TGF-β1+si-NC group， TGF-β1+si-MALAT1 group，TGF- β 1+si-MALAT1+anti-miR-150-5p group，and TGF-β1 group+si-MALAT1+CXCL14 group. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of MALAT1 mRNA， microRNA-150-5p （miR-150-5p）， and CXC chemokine ligand 14 （CXCL14） mRNA in serum and HSC of the subjects in various groups；Western blotting method was used to detect the expression levels of CXCL14， α-smooth muscle actin（α-SMA），and type Ⅰ collagen α 1 （COL1A1） proteins in serum of the subjects in various groups；CCK-8 method was used to detect the proliferation activities of the HSC in various groups； the expression levels of α- SMA，COL1A1 proteins in the HSC in various groups were detected by immunofluorescence；the targeting relationship between miR-150-5p and MALAT1 and CXCL14 3′-UTR genes was detected by dual luciferase reporting system. Results .The RT-qPCR results showed that compared with healthy group， the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in mild and severe liver fibrosis groups were increased（P<0.05）， while the expression levels of miR-150-5p were decreased （P<0.05）； compared with mild liver fibrosis group， the serum expression levels of MALAT1 mRNA and CXCL14 mRNA of the subjects in severe liver fibrosis group were increased （P<0.05），while the expression level of miR-150-5p was decreased （P<0.05）； compared with control group， the expression levels of MALAT1 mRNA and CXCL14 mRNA in the HSC in TGF- β1 group were increased （P<0.05）， while the expression level of miR-150-5p was decreased （P<0.05）； compared with TGF-β1+si-NC group， the expression levels of MALAT1 mRNA and CXCL14 mRNA in the HSC in TGF-β1+si MALAT1 group were decreased （P<0.05）， while the expression level of miR-150-5p was increased （P<0.05）； compared with TGF-β1+si MALAT1 group， the expression levels of miR-150-5p in the HSC in TGF- β1+si-MALAT1 and anti-miR-150-5p groups were decreased （P<0.05）， while the expression levels of CXCL14 mRNA were increased （P<0.05）； compared with TGF-β1+si-MALAT1 group， the expression level of CXCL14 mRNA in the HSC in TGF-β1+si-MALAT1+CXCL14 group was increased （P<0.05）.The Western blotting results showed that compared with healthy group， the serum expression levels of CXCL14 protein of the subjects in mild and severe liver fibrosis groups were increased （P<0.05）； compared with mild liver fibrosis group， the serum expression level of CXCL14 protein of the subjects in severe liver fibrosis group was increased （P<0.05）； compared with control group， the expression levels of α-SMA and COL1A1 proteins in the HSC in TGF-β1 group were increased （P<0.05）； compared with TGF-β1+si-NC group， the expression levels of α- SMA and COL1A1 proteins in the HSC in TGF-β1+si-MALAT1 group were decreased （P<0.05）； compared with TGF-β1+si-MALAT1 group， the expression levels of CXCL14，α-SMA and COL1A1 proteins of the HSC in TGF-β1+si-MALAT1+anti-miR-150-5p group and TGF-β1+siMALAT1+CXCL14 group were increased （P<0.05）.The CCK-8 method results showed that compared with control group， the proliferation activity of the HSC in TGF-β1 group was increased （P<0.05）； compared with TGF-β1+si-NC group， the proliferation activity of the HSC in TGF-β1+si-MALAT1 group was decreased （P<0.05）； compared with TGF-β1+si-MALAT1 group，the proliferation activities of the HSC in TGF-β1+si-MALAT1+anti-miR-150-5p group and TGF-β1+si-MALAT1+CXCL14 group were increased （P<0.05）. The immunofluorescence results showed that the expressions of α-SMA and COL1A1 proteins in the HSC in various groups was consistent with the results detected by Western blotting method. There was a targeting relationship between MALAT1 and CXCL14 3 '-UTR and miR-150-5p. The double luciferase reporter gene assay results showed that compared with miR-NC group， the luciferase activity of the HSC in miR-150-5p group co-transfected with MALAT1 WT or CXCL14 WT was decreased （P<0.05）. Conclusion Knocking down of MALAT1 can inhibit the activation of the HSC induced by TGF- β 1，and the mechanism may be related to the miR-150-5p/CXCL14 signaling pathway.

Objective To study the establishment and stability of hepatic cirrhosis animal model(CM). Methods The methods of compound factor, carbon tetrachloride(CCl₄) plus alcohol, corn flour and cholesterol plus alcohol were used to construct the hepatic cirrhosis rat models. 80 Wistar rats were selected, male and female were ,respectively. There were 40 Wistar rats treated with compound factor method.The fodder feed was 80% corn flour,20% animal oil, and 0.5% cholesterol, the only beverage was 5% alcohol .At the first time,the Wistar rats were administered with 40% CCl₄-olive oil mixture by subcutaneous injection (5 mL·kg_-1). Then they were hypodermically injected with 40% CCl₄-olive oil mixture (3 mL·kg^-1) every 3 days for six weeks. At the end of the 6th week,six Wistar rats were executed randomly and made into hepatic smears which were accorded with hepatic cirrhosis (The forming of pseudolobuli was the only standard to decide the successful establishment of CM). The rest rats were randomly divided into two groups. There were 13 Wistar rats treated with normal fodder feeding in each group. Meanwhile the rats in one group were hypodermically injected with 40% CCl₄-olive oil mixture (3 mL·kg^-1) every 7 days. Then the pathological changes at the end of 7th and 8th week were observed. Twenty Wistar rats treated with CCl₄ plus alcohol were fed with normal fodder food. Remain ing conditions were as same as those were used in the first 6 weeks of compound factor method. Twenty Wistar rats treated with corn flour and cholesterol plus alcohol weren′t hypodermically injected with CCl₄ and remaining conditions were same with those were used in the first 6 weeks of compound factor method.Results The rat models with hepatic cirrhosis treated with compound factor were established completely at the end of 6th week. Then the rats were fed with normal fodder food. Pseudolobuli were all disappeared after two weeks. And the rats were hypodermically injected with CCl₄ for 2 weeks after establishment of CM,pseudolobuli were mainly existed. Typical CM were not constructed with other two methods. Conclusion The forming rate of hepatic cirrhosis model constructed with compound factor is high. After establishment of the model, CCl₄ can keep the relative stability of the models.

Objective To observe the preventive effect of combination fish oil on pathological lesions in inner ear of diabetic rats and its mechanism. Methods Fifty male Wistar rats were randomly divided into three groups randomly :group A (normal control, n=16), group B (diabetic control, n=18), and group C (treatment group,n=16). Rats in group B and group C were injected intravenously with streptozocin(STZ) of 55 mg•kg^-1. Three weeks after injection,the rats in group C were continuously fed on combination fish oil everyday for 9 weeks. The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C) and very low density lipoprotein-cholesterol (VLDL-C) in blood of rats were detected.The animal auditory brain stem evoked potentials were recorded. Results The levels of TC,TG,LDL, and VLDL in group C descented obviously compared with group B (P<0.05), auditory brain stem evoked potentials wave Ⅲ was obviously shortened in latent period compared with group B (P<0.05) and the threshold of wave Ⅲ decreased (P<0.05). Mass of lipid droplet in hairy cells in inner ear, vacuolar degeneration of mitochondrion and Golgi′s apparatus, hypertrophy and hyperplasia of endothelial cells in micrangium, and thick basement membrane were observed under transmission electron microscopy in group B. But these pathological lesions did not appear in the rats in group C. Conclusion Combination fish oil can prevent the lesions of inner ear in diabetic rats.

Objective To establish a retroviral vector of small hairpin RNA (shRNA) expression in which the sense and antisense sequences targeting wild type human p53 were linked together with a 9-nucleotide loop to detect the interfering role of p53 gene in mammalian cells. Methods The human H₁ promoter was inserted into the upstream of the cytomegalovirus (CMV) promoter by using Xhol I and EcoR I sites. The resistanting hygromycin gene was replaced with the human CD₄ gene. These oligonucleotides were annealed and ligated the downstream of the H₁ promoter. All oligonucleotides were synthesized with Qiagen. HEK293 was infected with the shRNA vector and the expression of p53 was detected 72 h after infection by flow cytometry. Results The retroviral vector was successfully constructed and the expression of P53 protein was definitely suppressed in the HEK293 72 h after infection. Conclusion The applying of shRNA expression vector is possible to provide a prompt and promising method for evaluating the gene function of mammalian cells.

Objective To discuss the effects and mechanism of Rutongning capsule on hyperplasia of mammary glands in rats. Methods 50 female Wistar rats without pregnancy were randomly divided into normal group，model group，low dosage group，middle dosage group and high dosage group（n=10）. Estradiol and progesterone were used to induce hyperplasia models of mammary glands in rats. The rats with hyperplasia of mammary glands were administered by gavage with Rutongning. The diameter of the second twin nipples of the rats was measured and the structure of the mammary gland tissue was observed. The level of hormone in blood was detected by radioimmunity and posit ive cells of estrogen receptors（ER） and progesterone receptors(PR) were determ ined by immunohistochemical technique. Results Compared with model group, estradiol, prolactin, follicle stimulating hormone in middle and high dosage groups were decreased (P<0.05),and luteinizing hormone , progesterone were increased (P<0.05) ,and the diameter of the second twin nipples of the rats was reduced (P<0.05). Rutongning made a cini atrophy and make gland ducts, and their volumes became small significantly (P<0.01). Rutongning decreased the number of ER and PR positive cells significantly. Conclusion Rutongning could rectify the secretion of hormone in rats and weaken the biological effects of estrogen and progesterone on mammary glands，so the hyperplasia mammary glands could get recovery.

Objective To evaluate the efficacy of neoadjuvant chemotherapy (NAC) with nedaplatin, ifosfamide and peplomycin (SIP) for patients with locally advanced squamous cell carcinoma of cervix, and to determine whether or not the mitotic index and the Ki-67 labeling index (LI) obtained from biopsy specimens were useful as predictors of NAC response. Methods Twelve cases (stage Ib:3 and II:9) of cervical carcinoma with more than 4 cm in diameter were treated with 1 to 2 courses of SIP therapy followed by surgery. The response to SIP was measured by magnetic resonance imaging (MRI).　The relationship of response and serum SCC, and the changes of mitotic index (MI) and the Ki-67 LI in the biopsy specimens were investiged before and after SIP therapy. Results Seven cases resulted in partial response (PR) and 5 cases did not change (NC), the effective rate was 58.8%. In cases with PR, tumor size reduced markedly after the first course of SIP and serum SCC decreased. MI values in PR cases decreased, and Ki-67 LI was unchanged. On the other hand, in NC cases, MI values did not decrease and Ki-67 LI tended to increase. Conclusion NAC with nedaplatin, ifosfamide and peplomycin was effective for patients with locally advanced squamous cell carcinoma of cervix. Alterations of MRI, serum SCC, MI and Ki-67 LI can be predictors for NAC.

Abstract:Objective To observe the therapeutic effect of Simiaoyong’an Decoction (SMYA) on the rat models with thromboangiitis obliterans (TAO),and to explore the action mechanism.Methods The rats were randomly divided into 6 groups:sham operation group,model group,mailuoning group(MLN 2.7 g/kg),high,medium and low doses of SMYA groups (24.3,16.2,8.1 g/kg). The  TAO model was prepared by injecting sodium laurate into the femoral artery of rats.The rats were administered by intragastric administration after prepared model.After 15 d administration,the pathological changes of blood vessel and surrounding tissue were observed with HE staining,the contents of IL-6,TNF-α,CRP,TXB2 and 6-K-PGF1α in plasma were measured by enzyme-linked immunosorbent assay (ELISA),and the content of NO in plasma was measurd by ultraviolet spectrophotometer.Results Compared with model group,the contents of plasma IL-6,TNF-α,TXB2 and CRP in high dose of SMYA(24.3 g/kg) group were significantly decreased,and the contents of NO and 6-K-PGF1α were markedly increased (P<0.05 or P<0.01);the contents of plasma TNF-α in medium and low doses of SMYA (16.2 and 8.1 g/kg) groups were significantly decreased (P<0.01).The  vascular endothelial cells in model group were badly damaged with a large number of  inflammatory cells infiltration,and the  vascular endothelial cells in high dose of SMYA group showed integrity with  little inflammatory cells infiltration.Conclusion SMYA has a good therapeutic effect on rat model of TAO,and the mechanism may be related to anti-inflammatory and  maintaining the balance function of TXA2 and PGI2 in plasma.

Objective To explore the protective effects of egg-milk with metallothionein (MT) on mice damaged by irradiation. Methods After the mice were irradiated with 2.5 Gy (12.5 mGy·min^-1) X-rays, they were fed with egg-milk with MT for 14 days. The level of WBC in peripheral blood, lymphocyte proliferation rate, DNA content in marrow cells, malondiadehyde (MDA) levels and the activities of antioxidant enzymes (SOD, CAT and GSH-Px) in liver and kidney were measured. Results After the irradiated mice were fed with egg-milk with MT, especially in the egg-milk with MT of mild and high doses plus irradiation groups, the WBC number, lymphocyte proliferation rate and DNA content of marrow cells in the mice were significantly higher than those in the irradiation group (P<0.01), at the same time, the percentage of thymocyte G0/G₁ phase of these mice were significantly lower than that of the irradiation group (P<0.01), and the percentage of thymocyte S and G₂/M phase were signifi cantly higher than tho se in the irradiation group　(P<0.01 or P<0.05). Compared with the no rmal control, MDA levels in liver and kidney of the irradiation group increased significantly (P<0.01 or P<0.05), and the antioxidant enzyme activit ies of SOD, CAT and GSH-Px decreased significantly (P<0.01 or P<0.05). While in the egg-milk with MT plus irradiation groups, the MDA contents were significantly lower than those in the irradiation group (P<0.01) and the activities of SOD, CAT and GSH-Px were significantly higher than those in the irra diation group (P<0.01 or P<0.05). Conclusion The egg-milk with MT has protective effects on mice injuried by irradiation. Key words：metallothionein/pharmacology; radiation injuries, e xperimental;malondialdehyde; DNA

Objective To study the effects of muskrat and moschus of anti-inflammation and annalgesia. Methods The effects of muskrat and moschus of anti-inflammation were observed by using torsive-body method. The effects of muskrat and moschus of analgesia were observed by mice inflammation models caused by dimethylbenzene and mice models with reinforce permeability of abdominal cavity capillary caused by acetic acid. Results Both muskrat and moschus could significantly reduce the times of torsive-body in mice caused by glacial acetic acid (P<0.05 or P<0.01). And they obviously inhibited ear swelling of mice caused by dimethylbenzene (P<0.05 or P<0.01). The effects of moschus were more obvious than muskrat when they were given with the same dosage. Muskrat and moschus had obvious inhibitory effects on mice inflammation of reinforce permeability of abdominal cavity capillary caused by acetic acid (P<0.05 or P<0.01).They had no difference of effects when they were given with the same dosage（P>0.05）. Conclusion Muskrat and moschus have the same effects of anti-inflammation and analgesia.

To investigate the influence of genistein(Gen) on the growth  and apoptosis regulation of hepatoma carcinoma cells SMMC-7721.Methods SMMC-7721 cells were divided into three Gen groups treated with different concentrations(5.0,10.0,20.0 mg·L^-1）and control group(without Gen).After SMMC-7721 cells were treated for 24 and 48 h,the changes of ultrastructure of cells were observed under electron microscope, MTT was used to detect the inhibitory rate of proliferation of SMMC-7721 cells, flow cytometry was performed to analyze cell phase;immunohistochemistry was used to detect the expression of Caspases-3 protein.Results Compared　 with control group,chromatin in cellular nucleus became conglomeration or lumping,with chondriosome swelling in Gen groups. With the increasing of Gen concentration,the cellular nucleus became condensed,heterochromatin agglutinating,ground cytoplasm obviously cavitating;with the increase of Gen concentration and prolongation of  time,from 24 to 48 h, MTT showed the inhibitory rate of  proliferation  of SMMC-7721 cells  increased  in a time- and dose-dependent  manner.Flow cytometry analysis showed that with the increase of Gen concentration,the number of cells at G₂/M  phase increased,the number of cells at S phase decreased and the ratio of G₀/G₁ also decreased.Immunohistochemistry results showed the expression of protein Caspases-3 increased with the Gen concentration in a dose
-dependent manner（P<0.01）.
Conclusion Gen has significantly  inhibitory effect on the growth of hepatoma carcinoma cells SMMC-7721,up-regulating the expression of Caspases-3 protein and inducing  apoptosis may be one of the mechanisms.

Objective To amplify the enamel matrix serine proteins 1(EMSP1) in vitro and to establish gene targeting vector and transgenic mouse models to study the physiological and pathological function of EMSP1.Methods Two pairs of primers were d esigned by Oligo 6.0 software and synthesized according to EMSP1 gene sequence. Two gene fragments of 129 strain mouse EMSP1 were amplified from mouse genomic DNA by PCR-5 000 bp 5′ arm and 1 700 bp 3′arm. The two arms were cloned into a universal gene knockout vector-pSSC-9 respectively on either side of the positive selective marker neo. The targeting vector was identified by PCR， restriction endonuclease and sequence analysis. Results The restriction endonuclease identifiation result indicated that 5 000 bp and 1 700 bp fragments were obtained and homogeneous long and short arms were cloned into vector.DNA squence analysis showed that EMSP1 gene targeting vector pSSC-9-EMSP1 was successfully constructed. Conclusion The mouse EMSP1 gene targeting vector is constructed successfully.

Objective To study the effect of fluoride on the expressions of collagen typeⅠ(ColⅠ) and collagen type Ⅲ (Col Ⅲ) in the myocardium of rats.Methods The rat model was established by administration of different concentration o f sodium fluoride. The myocardium of rats were stained by picrosirium red，observed by polarization microscopy，analyzed by image analysis system. The expressions of ColⅠand Col Ⅲ in myocardium of rats were determined by immunohistochemistry method.Results The percentages of ColⅠand Col Ⅲ area in the myocardium of rats treated with different concentration of sodium fluoride were a l ittle higher than those of control group.No significant difference was seen among different groups of the expressions of ColⅠand Col Ⅲ（P＞0.05）.The expressions of ColⅠand Col Ⅲ protein were ranged between 0-10%.There was no significant difference among different groups either（P＞0.05）. Conclusion Chronic fluorosis has no obvious effect on myocardial collagen metabolism of rats and myocardial collagen is not likely the main target of fluoride.

Objective To study the effect of plyrhachis vicina rogher (PVR) on immune function in mice with tumor and the mechanism of tumor inhibition. Methods The BALB/c mice were randomly divided into normal control (NC) group and experimental groups including high, middle and low doses of alcohol extract of PVR. The mice were inoculated with S₁₈₀ cell of ascites sarcoma in mice. The tumor model of animal was established. NC group was fed on distillative water. Experimental groups were fed on 0.5,1.0, and 1.5 g•kg^-1 alcohol extract of PVR for 10 days. The tumor weight,inhibitory rate of tumor,indexes of spleen, IL-2,activities of tumor killer LAK and NK,proliferation of T,B lymphocytes were used as evaluated indexes. Results Spleen weight, killer activities of the LAK and NK, proliferous reaction of the spleen cell and the level of IL-2 secretion in mice treated with alcohol extract of PVR were significantly higher than those in NC group. Conclusion PVR may promote specific and nonspecific immune function of organism and inhibit tumor cells indirectly.

Objective To discuss the diagnosis and treatment of one patient with ruptured intracranial aneurysm complicated with acute myocardial infarction（AMI）， and to provide the reference for the clinical diagnosis， treatment， and anesthesia of the disease. Methods The clinical data， imaging findings， and anesthesia methods of one patient with ruptured intracranial aneurysm complicated with AMI were retrospectively analyzed，and the analysis was performed combined with the relevant literatures. Results The patient was admitted to hospital due to sudden severe headache with nausea and vomiting for 4 h. A total of 1 h and 10 min after admission，the multi-slice CT results showed subarachnoid hemorrhage（SAH）.There was little bilateral ventricular effusion， and the intracranial angiography results showed a tumor in the posterior communicating segment of the right internal carotid artery.A total of 2 h 52 min after admission，the myoglobin was 483.6 μg·L^－1，the troponin I was 4.990 μg·L^－1，the creatine kinase isoenzyme-MB（CK-MB） was 45.70 μg·L^－1.A total of 16 h 31 min after admission， the ECG results showed sinus bradycardia， left ventricular hypertrophy， and ST-T segment changes. The initial diagnosis of the patient was SAH， AMI， and hypertension grade 3 （very high risk）. After early comprehensive treatment， the patient underwent emergency clipping of cerebral aneurysm after 3 d.The anesthesia method was tracheal intubation ansthesia，and the anesthetic drugs were carefully selected to achieve the best blood flow and anesthesia effect.The vital signs of the patient were stable during the operation， and the condition of the patient was improved and discharged after 7 d. Conclusion For the patients with intracranial aneurysm rupture complicated with AMI，CT， intracranial angiography， and myocardial markers are the important examinations for the diagnosis and differential diagnosis； controlling the blood pressure is the key point for the treatment and anesthesia.

Abstract:Objective To investigate the genetic assaciation between the estrogen receptor-alpha gene polymorphism and uterine leiomyoma in Northern Han Chinese.Methods The method of PCR-RFLP was conducted to examine the genotypes of rs722209 and rs9322346 of estrogen receptor-alpha gene in 337 subjects,including 167 uterine leiomyoma and 170 controls.The association between the polymorphisms and uterine leiomyoma was analyzed with SPSS 12.0.Results The genotypic frequencies of the 2 SNPs of estrogen receptor-alpha gene did not deviate from Hardy-Weinberg equilibrium in both patient and control groups.The allelic and genotypic frequencies of the 2 SNPs had no remarkable difference between patient and control groups(P>0.05).Conclusion The gene polymorphisms of rs722209 and rs9322346 of estrogen receptor-alpha may be not associated with uterine leiomyoma.

Objective To study the effects of Gross Saponin of Tribulus Terrestris (GSTT) on acute myocardial infarction induced by ligating coronary artery in anaesthetic thoraco-opening dogs by injection. Methods Thirty domestic dogs were randomly divided into 5 groups (6 in each group): blanking control group, positive control group(Xuesaitong), GSTT groups with doses of 6.26 , 12.52, and 25.0 mg•kg^-1, respectively. The descending branch of left coronary artery was ligated to construct the model of acute myocardial infarction. The effects of GSTT on epicardium electrocardiogram(EECC), myocardial infarction range(MIS) , and serum myocardium enzyme were observed. Results The ischemia degree(Σ-ST, P<0.05) and the ischemia area(N-ST, P<0.05) were reduced in GSTT groups with various doses compared with controls. In addition, GSTT reduced the serum myocardium enzyme levels　(P<0.01) and myocardial infarction ranges(P<0.001) compared with controls. Conclusion GSTT can significantly protect ischemic myocardium of the anesthetic thoraco-opening dogs mediated by ligating coronary artery.

Objective  To establish stable and reliable rat C6 glioma model,and to perform MRI dynamic observation and pathomorphological observation in model animal brain,and to provide experimental basis for pharmaceutical research on　anti-glioma drugs.Methods The C6 glioma cells were cultured and 20 μL cultural fluid containing 1×10⁶ C6 cells was sterotactically implanted into the left caudate nuclei in 10 male Wistar rats,respectively.The changes in the behavior of the rats after implantation were observed and recorded.MRI dynamic scanning was performed in 10 rats  2,3 and 4 weeks after implantation and the brain tissues were taken for general and pathological examination when the 10 rats were naturally dead.The survival period of tumor-bearing rats was calculated.Results 2 weeks after implantation the rats showed decreased activities and food intake,fur lackluster,and conjunctival congestion a
nd so on;3 weeks later,some rats appeared nerve symptoms such as body twitch,body hemiplegy,body distortion,rotation and so on.All the 10 rats died in 8 -
30 d.The median surrival period of the tumor-bearing rats was 18 d,the average survival period was (18.3±7.3) d.The pathological examination showed that
the tumor cells were arranged irregularly and closely and karyokinesis was easy to see;tumor vascular tissue proliferation and tumor invasive growth into
surrounding noral tissues were found.The expression of glial fibrillary acidic protein(GFAP) was positive in the tumors.
Conclusion  A stable animal model of  intracranial glioma is sucessfully established by  stereotactic implantation of C6 cells into the rat caudate nucleus.The results of  MRI 〖JP2〗dynamic observation and pathohistological observation on the model animal brain tissue.can provide experimental
basis for selecting the appropriate time window to perform the pharmaceutical research on anti-glioma drugs.

Methods The clinical manifestations of one patient with ITTC were observed.The general morphology of the tumor was observed with naked eyes， laboratory examination and imaging examination were performed， the fine needle aspiration cytology of the thyroid tumor was observed， the morphology of the tumor tissue was observed under light microscope， immunohistochemistry staining and in situ hybridization were performed in the tumor tissue， and the patient was followed up 3 and 11 months after operation；the relevant literatures were reviewed. Results The main clinical manifestation of the ITTC patient was a slightly hard mass in the thyroid.The fine needle aspiration cytology results showed that the tumor cells showed a three-dimensional structure，and the prominent nucleoli could be seen in some cells. The histology results showed that the cancer cells were nest like or island like and differentiated into the squamous cells； the fibrous tissue proliferated accompanying with lymphocyte infiltration around the cancer cells and the right cervical lymph node metastasis. The immunophenotype detection results showed that the cancer cells expressed CD117， CD5 and P63， but didn’t express TdT and TTF-1，EBER-ISH （－）. The pathological diagnosis was ITTC accompanying with cervical lymph node metastasis. No recurrence or metastasis was found on CT 3 and 11 months after operation. Conclusion ITTC has low malignancy degree，which can relapse and metastasize，and combined with surgery and radiotherapy， the prognosis of the patients is good. Objective To study the clinicopathological features， treatment method and prognosis of intrathyroid thymic carcinoma （ITTC），and to strengthen the clinicans’ understandings of the disease.

Objective To investigate the clinical diagnostic value of bone marrow prostate specific antigen (PSA) in bone metastasis of prostatic cancer. Methods Twenty cases of prostatic cancer were divided into positive bone scanning group (n=12) and negative bone scanning group (n=8) according to the results of isotope bone scanning,and 20 cases of benign prostatic hyperplasia were used as control.The levels of PSA in bone marrow and serum were measured in all patients to study the correlationship of bone marrow PSA and bone metastasis, and the ratio of bone marrow PSA to serum PSA was calculated. Results The level of PSA in bone marrow in positive bone scanning group were statistically higher than those in negative bone scanning group and control group (P<0.05)). The diagnostic ratio of bone marrow PSA to serum PSA had been set to 0.70, the sensitivity to predict bone metastasis was 78%, the specificity was 86%, the accuracy was 73%. Conclusion The level of PSA in bone marrow has close correlation with bone metastasis of prostatic cancer and it is a marker of bone metastasis of prostate cancer. The ratio of 0.70 of bone marrow PSA to serum PSA can be used as a index to diagnose bone metastasis.

Methods The levels of D-Dimer and tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) were detected in 92 middle-aged patients with nonvalvular atrial fibrillation (AF group) and 60 patients with sinus rhythm (control group) by immune turbidimetry and enzyme linked immunoadsorbent assay (ELISA). Univariate analysis was used to determine the ifferences between two groups, and covariance analysis was used to determine the factors which might affect coagulation and fibrinolysis indexes. Results ①The plasma levels of D-Dimer ［(0.16±0.10）mg·L^-1］ and t-PA ［(42.58±30.28）μg·L^-1］ and PAI-1 ［(86. 03±21.43)μg·L^-1］ in AF group were significantly higher than those in the control group ［(0.10±0.08)mg·L^-1，(26.02±13.84)μg·L^-1， (64.94±24.35)μg·L^-1］(P<0.05 or　P<0.001). The ratio of PAI-1 /t-PA in AF group was higher than that in control group slightly. ②After adjustment of the factors which included sex, age and plasma reatinine, uric acid, blood sugar, triglyceride and cholesterol, the levels of D-Dimer（P=0.047）, t-PA（P=0.264）and PAI-1（P=0.001）in AF group were higher than those in the control group. Conclusion The middle-old aged patients with nonvalvular atria l fibrillation lose their balance of coagulation and fibrinolysis in the state of hypercoagulated and hypofibrinolysis.

Objective: To investigate the effect of dapagliflozin on the liver function of the patients with type 2 diabetes mellitus (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD), and to analyze its improvement effect on liver injury of the T2DM patients. Methods: The clinical data of 68 T2DM patients complicated with NAFLDS who did not receive any treatment were retrospectively analyzed, including 30 patients in acarbose group and 38 patients in dapagliflozin group, and the patients in two groups received acarbose 150 mg·d^-1+ metformin 2 000 mg·d^-1 and dapagliflozin 10 mg·d^-1 + metformin 2 000 mg·d^-1 treatment for 24 weeks, respectively. The general data of the patients before and after treatment were collected.The fasting venous blood of the patients in two groups was collected and the serum biochemical indexes and liver function indexes were detected.The serum biochemical indexes included fasting blood glucose(FBG) and glycosylated hemoglobin (HbA1c),and the homeostasis model assessment 2-insulin resistance index(HMOA2-IR) was calculated.The liver function indexes included aspartic transaminase (AST), alanine aminotransferase (ALT), glutamyltranspeptidase (GGT), alkaline phosphatase (ALP), total bilirubin (TBIL),and direct bilirubin (DBIL);the non-alcoholic fatty liver disease fibrosis score (NAFLDFS) was calculated.The each detection index of the patients in two groups was compared and analyzed. Results: Compared with before treatment, the levels of FBG, HbA1c, AST, GGT, and ALP and HOMA2-IR in acarbose group after treatment were significantly decreased(P<0.01) and the levels of TBIL was significantly increased (P<0.01). Compared with before treatment, the levels of FBG, HbA1c, AST, ALT, GGT, and ALP and HOMA2-IR,NAFLDFS in dapagliflozin group were significantly decreased after treatment (P<0.01) and the levels of TBIL and DBIL were significantly increased (P<0.01). The levels of FBG, AST, ALT, GGT, and ALP and NAFLDF in dapagliflozin group after treatment were significantly decreased compared with acarbose group (P<0.01). Conclusion: Dagliflozin can improve the liver function of the patients with T2DM complicated with NAFLD, and its mechanism may be related to the effect of dapagliflozin decreasing the blood glucose and body weight.

Objective To elucidate the effects of Diemailing injection (DMLI) on organ blood flow and vascular resistance in anesthetized dogs. Methods Cerebral blood flow，cerebral vascular resistance，peripheral blood flow,peripheral vascular resistance,blood pressure, and heart rate (HR) were measured using the MF-27 electromagnetic flowmeter. Results DMLI(4 and 8 mL•kg^-1 iv drip) increased cerebral blood flow and decreased cerebrovascular resistance within 180 min，its effect was more obvious than Mailuoning injection (MLNI).DMLI also elevated peripheral blood flow and lowered peripheral vascular resistance,its effect was less than MLNI.HR was slowed down only at 60 min whereas at other time not.DMLI obviously reduced diastolic pressure but exerted no effect on systolic pressure. Conclusion Protective effect of DMLI on experimental cerebral ischemia may be related to improving cerebral circulation and increasing cerebral blood supplyment.

Objective To study the effects of IL-24 gene combined with ionizing radiation on apoptosis in PC-3 cell line in order to prepare the ground for combined therapy of IL-24 gene and ionzing raditaion for tumor.Methods The 　experiment was divided into sham irradiation group and irradiation groups with different irradiation doses,which were 2,4,6,8,12 and 18 Gy, respectively.To detect the expression of IL-24 gene,three groups were included,which were control group (1×PBS),vector group and IL-24 gene group.To detect the apoptotic effect of IL-24 gene combined with ionization radiation on PC-3 cell line,the experiment was divided into control group,vector group,IL-24 gene group,irradiation group (6 Gy),vector combined with irradiation group and IL-24 gene combined with irradiation group.Alkaline lysis assay was used to extract and purify the plasmid.Plasmids were transfected into PC-3 cell line by polyethyleneimine（PEI）in vitro.The expression of the interest gene was detected by RT-PCR.The changes of apoptosis in PC-3 cell line were detected by flow cytometry（FCM）using the staining of Annexin-V and PI.Results Compared with sham-irradiation group,the apoptotic percentage of PC-3 cell line did not show marked change after 2 and 4 Gy X-rays irradiation for 48 h（P>0.05）.The apoptotic percentage was increased significantly after X-rays irradiation with the dose of 6 Gy（P<0.01），and the mean apoptotic percentage of PC-3 cell line was increased by a factor of 1.6 to 3.0 compared with sham-irradiation group.The expression of IL-24 gene could be observed in PC-3 cell line transfected by IL-24 gene except in control group and vector group，and all of them showed the expression of GAPDH gene.As compared with the other groups,the number of early apoptotic cells of PC-3 cell line in the IL-24 gene combined with irradiation group was increased significantly （P<0.05） except in irradiation roup.The number of late apoptotic and necrotic cells of PC-3 cell line in the IL-24 gene combined with irradiation group was increased significantly compared with control group,irradiation group and vector combined with irradiation group（P<0.05,P<0.01）.As compared with the other groups,the total number of apoptotic and necrotic cells in the IL-24 gene combined with irradiation group was increased significantly （P<0.05，P<0.01） except in IL-24 gene group.Conclusion IL-24 gene combined with X-rays irradiation could induce effective apoptosis in PC-3 cell line.

Abstract:Objective To study the effect of low dose cyclophosphamide on function of CD4⁺CD25⁺Treg of mice with Lewis lung cancer and investigate the relationship between CD4⁺CD25⁺Treg and the genesis and development of tumor.Methods The models were divided into three groups:CTX group,tumor group and control group.In CTX group, models were established by injection CTX (25 mg•kg^-1) and subcultivated Lewis lung cancer cells were injected subcutaneously to the right axilla of C57BL/6 mice 7 d later.The dynamic changes of tumor volume were observed.The changes of the number of CD4⁺CD25⁺Treg were detected by flow cytometer and the expression of Foxp3 mRNA in spleen was analyzed by semi-quantitative RT-PCR.The changes of T lymphocyte proliferation in spleen were detected by MTT test.The changes of T lymphocyte killing function in spleen were detected by LDH releasing test.Results Compared with tumor group,CTX treatment might delay the development of tumor in 19 d-mice with Lewis lung cancer.The number of CD4⁺CD25⁺ Treg in spleen of mice in CTX group was lower than that in tumor group (P<0.05).The expression of Foxp3 mRNA in spleen lymphocyte in CTX group was significantly lower than that in tumor group (P<0.05).The changes of T lymphocyte proliferation in spleen in CTX group were significantly higher than that in tumor group (P<0.05).The changes of T lymphocyte killing function in spleen in tumor group was not significantly lower than that in CTX group (P>0.05). Conclusion After CTX injection,the number of CD4⁺CD25⁺Treg and the expression of Foxp3 mRNA in spleen of mice with Lewis lung cancer decrease,the immunological function of mice with Lewis lung cancer increase.It can provid experiment evidence for the tumor immunotherapy aimed directly at CD4⁺CD25⁺Treg.

Objective To investigate the effects of pancreatic kininogenase on pressure,myocardical fibrosis and serum nitric oxide (NO) level in spontaneously hypertensive rats (SHR). Methods Twenty-four (fifteen weeks) male SHR were randomly divided into three groups:SHR group,pancreatic kininogenase group and captopril group（n=8 ），8 male Wistar kyoto rats with normal blood pressure were considered as control group. Pancreatic kininogenase was given by peritoneal injection (7.2 U•kg^-1•d^-1), captopril was given by intragast ric administration（10 mg•kg^-1•d^-1）, the rats in SHR group and rats with normol blood pressure in control group were treated with 0.9% NaCl（2 mL•kg^-1•d^-1）administered through peritoneal injection. After four-week experiment, the pressure was measured in rats througth carotid artery,then the rats were sacrificed , and LVMI,CVF,PVCA and serum NO level were measured. Results In SHR group, SBP,LVMI,CVF and PVCA were higher, serum NO level was decreased obviously than those in control group (P<0.05). After treatment of pancreatic kininogenase,the serum NO level raised obviously,other indexes obviously reduced than those in SHR group (P<0.05),but except SBP,there were no significant difference compared with captopril group (P>0.05). Conclusion Pancreatic kininogenase can obviously reduce the blood pressure and reverse myocardial fibrosis,its mechanism may be concerned with the increasing of NO level in SHR.

Objective To explore the inhibitory effects of extracts from Trapa manshurica Fler. on liver cancer cells in vivo and in vitro. Methods The sensitivity of extracts from Trapa manshurica Fler. to liver cancer cells in vitro was determined with ³H-TdR incorporation method. The inhibitory rate and growth of the tumor and the weight change of the mice were observed. Results When the dosage was 225.00 g•L^-1, the inhibitory rate of tumor was 75.49% in vitro and the inhibitory rate of tumor was 60% in vivo. Conclusion The extracts from Trapa manshurica Fler. has the inhibitory effects on tumor.

Objective To explore the effects of glutamine on hepatocytes apoptosis in rats with obstructive jaundice. Methods 50 male Wistar rats were randomly divided into three groups:control(SL，n=10),obstructive jaundice group (OJ，n=20) and glutamine treatment group (TG，n=20).The hepatocytes apoptosis and the expressions of Bcl-2 and Bax were detected by using TUNEL and immunohistochemistry method. Results The expression of Bcl-2 in rat hepatocytes was increased 2 weeks after treatment in TG group. It expressed positively (+++). The expression of Bax was also decreased compared with the SL and OJ groups in two weeks. It expressed positively (++). Apoptotic index was 15.75±3.68. The difference was significant (P<0.05). Conclusion The glutamine can decrease hepatocytes apoptosis,the expression of Bax, and increase the expression of Bcl-2.

Objective To detect the expressions of hypoxia inducible factor - 1α (HIF-1α) and microvesseldensity (MVD)in ovarian epithelial tumor tissues ,and investigate the effect of HIF-1α and MVD in development of ovarian epithelial tumor.Methods 237 patients with ovarian epithelial tumor were selected,including 79 cases of serous cystadenoma, 12 cases of borderline serous cystadenoma,51 cases of serous cystadenocarcinoma, 67 cases of mucinous cystadenoma,9 cases of borderline mucinous cystadenoma and 19 cases of mucous cystoadenocarcinoma.Serous cystadenocarcinoma and mucous cystoadenocarcinoma were divided into different stages according to FIGO operation-pathology,stage Ⅰ was 13 cases,stage Ⅱ 26 cases,stage Ⅲ 19 cases and stage Ⅵ 12 cases.The expressions of of HIF-1α and MVD in ovarian epithelial tumor tissues were detected with immunohistochemistry.The expressions of HIF-1α and MVD were compared in benign tumor,borderline tumor and malignant tumor,as well as in different stages of malignant tumor.The relationship between the expression of HIF-1α and MVD value was also analyzed.Results The positive expression rate of HIF-1α in borderline serous cystadenoma group was significantly increased,compared with ovarian benign tumors (serous cystadenoma and mucinous cystadenoma groups)(P<0.05).The positive expression rates of HIF-1α in ovarian serous cystadenocarcinoma and mucous cystoadenocarcinoma were significantly superior to the benign tumors(P<0.05).The expressions of MVD in borderline serous cystadenoma and borderline mucinous cystadenoma groups were significantly higher than that in ovarian benign tumors(P<0.05).The expressions of MVD in serous cystadenocarcinoma and mucous cystoadenocarcinoma groups were sinigicantly higher than those in benign tumor group and borderline tumor group(P<0.05).The positive expression rates of HIF-1α and MVD were raised up with the process of the stage of FIGO operation-pathology.And there was positive correlation between HIF- 1α positive expression and MVD (r=0.540，P<0.01).Conclusion The overexpression of HIF-1α in ovarian epithelial tumor is closely related with angiogenesis,which may play an important role in the occurrence and development of ovarian epithelial tumor.

Objective To study the condition of pheochromocytoma cells (PC12 cells) differentiation into neurons induced by nerve growth factor (NGF). Methods There were five groups :10,20,50, and 80 μg•L^-1 NGF groups and control group which contained 10% fetal calf serum. PC12 cells were exposed to NGF with different concentrations for 72 h and the lengh of neurite and max diameter of PC12 cells were observed at 24,48,and 72 h, respectively. The expression of MAP2 was detected by immunocytochemistry after PC12 cells were exposed to NGF for 72 h . Results The number of MAP2 positive cells was significantly increased in 20, 50, and 80 μg•L^-1 groups compared with control group and the most significant concentration was 50 μg•L^-1（P<0.05）. The length of total primary neurite and max diameter of PC12 cells in 20,50, and 80 μg•L^-1 NGF groups were obviously longer and bigger than those in control group and the most significant concentration also was 50 μg•L^-1（P<0.01）. Conclusion PC12 cells could differentiate into neurons induced by NGF, and 50 μg•L^-1 is the optimal concentration in which NGF could induce PC12 cells to differentiate into neurons.

Abstract:Objective To investigate the methods of isolation，culture，identification and labeling of mesenchymal stem cells(MSCs) in vitro　and lay a foundation for further study on intervention of MSCs on immunologic rejection of organ transplantation. Methods MSCs were isolated and cultivated by adherent methods . The expressions of CD90 and CD45 of cells were analyzed by using flow cytometry in order to identify MSCs．The third generation of MSCs were labeled by DAPI,the labeling efficiency was detected．Results Primary cultured MSCs adhered to plastic surface within 48 h and reached 90％ confluence within 7－10 d ．Flow cytometry showed that the positive rates of CD90 and CD45 of MSCs at third generation were 99.8% and 6.8%. MSCs expressed CD90 but no CD45．All of the MSCs after labeling by DAPI showed blue fluorescence by immunofluoroscope. DAPI labeling was sensitive and highly efficient to MSCs．Conclusion Adherent method is simple and easy to isolate and cultivate MSCs and it can serve as a routine method．DAPI labeling can be used as a efficient method to label MSCs．

Abstract:Objective To explore the association between the C46T gene polymorphism in the exon-1 region of the coagulation factorⅫ(FⅫ) and venous thrombosis. Methods PCR-RFLP method was used to investigate the distribution frequency of the C46T gene polymorphism in the 87 venous thrombosis patients and 129 healthy controls.The relation of the C46T gene polymorphism with venous thrombosis was analyzed by Logistic un-conditional regression.Results There were three kinds of genotypes including 46C/C,46C/T,46T/T in the study population.The distribution frequency of 46C/C genotype in the venous thrombosis patients (6.9%) was higher than that in the healthy controls(4.65%).There was no relativity between 46C/C genotype and venous thrombosis.（χ²=1.036，P=0.309，OR=1.86，95%CI：0.57—6.12）.Conclusion It is not deemed that the C46T gene polymorphism of FⅫ is an independent risk factor in the venous thrombosis and the relativity between the deep venous thrombosis and C46T gene polymorphism is going on research.

Abstract:Objective To explore the expression of beta 2 adrenergic receptors(β₂-AR) in bone marrow cells and its significance and provide a scientific basis for the theory of hematopoietic regulation by nerve factors. Methods After　 bone marrow mononuclear cells were isolated from normal mouse bone marrow by the Ficoll density gradient centrifugation，the expressions of β₂-AR were studied by immunohistochemistry in bone marrow mononuclear cells,cultured bone marrow stromal cells,GM-CFUs,paraffin sections and smears of bone marrow.Mouse models of immune-mediated aplastic anemia(AA) were made by lymphocyte infusion.Mice were divided into 3 groups including animal model group,β₂-AR antagonist (ICI 118551) group,and control group,and the expression of β₂-AR in bone marrow mononuclear cells at the 8 th day was dete cted by Western blotting.Results The　 positive expression of β₂-AR in cytoplasm or membrane of normal mouse bone marrow cells showed different brown by immunohistochemical staining.There were intensive positive expressions of β₂-AR in mononuclear cells(48%) and bone marrow stromal cells (68%),while progenitor cells only showed middle-positive(55%)or weak positive(45%),and other hematopoietic accessory cells were positive in varying degrees.The relative values of β₂-AR of bone marrow mononuclear cells in 3 groups were 1.03±0.31,1.72±0.29 and 1.41±0.35,respectively;the expression of β₂-AR in animal model group was more lower than that in control group（P<0.05）,and the expression of β₂-AR in β₂-AR antagonist group was more higher than that in control group（P<0.05）. Conclusion β₂-AR can express in bone marrow cells of normal mice, and the expression level of β₂-AR is related to hematopoietic function of bone marrow,it may be involved in hematopoietic regulation by neuro-endocrine-immunity net work.

Objective To discuss the effect of ghrelin on the differentiation of the adipose mesenchymal stem cells （ADSCs） into the neurons，and to clarify its possible mechanism. Methods The ADSCs were randomly divided into blank group， neural differentiation inducer group， and ghrelin group （given 600 μg·L^－1 Ghrelin）， U0126 group （given 40 ng·L^－1 U0126）， Ghrelin+U0126 group （given 600 μg·L^－1 Ghrelin+40 ng·L^－1 U0126），and Ghrelin receptor blocker D-Lys3-GHRP-6 （D-Lys3-GHRP-6） group （given 10^－10 g·L^－1 D-Lys3-GHRP-6）. The pathomorphology of the cells in various groups was observed under inverted microscope；the positive expression rates of neurofilament protein （NF）-200 and tubulin （Tuj-1） in the cells in various groups were detected by immunofluorescence method； the expression levels of the mitogen activated protein kinase （MAPK）/extracellular signal-regulated kinase （ERK） pathway proteins，neuron specific nuclear protein （NeuN）， Tuj-1， neuron specific enolase （NSE），NF，growth hormone secretagogne receptor（GHSR），fetal liver kinase 1（Flk1）， and CD29 proteins in the cells in various groups were detected by Western blotting method. Results On the 10th day after cell culture， the ADSCs in blank group showed the fusions of long spindle and spiral shapes； the body of the cells in neural differentiation inducer group contracted，and the long spindle like cell body was transformed into the round-like shape， the protrusion length extended， and the number of neuron-like cells was increased； in Ghrelin group， the number of body protrusions of the cells was increased and the number of round-like like cells was increased； there was a small amount of cell body contraction and circular transformation in U0126 group and D-Lys3-GHRP-6 group. The immunofluorescence assay results showed that compared with blank group， the positive expression rates of NF-200 and Tuj-1 in the cells in neural differentiation inducer group were increased （P<0.05）； compared with neural differentiation inducer group， the positive expression rates of NF-200 and Tuj-1 in the cells in Ghrelin group were increased （P<0.05）， while the positive expression rates of NF-200 and Tuj-1 in the cells in U0126 group and D-Lys3-GHRP-6 group were decreased （P<0.05）； compared with Ghrelin group， the positive expression rates of NF-200 and Tuj-1 in the cells in U0126 group， D-Lys3-GHRP-6 group， and Ghrelin+U0126 group were decreased （P<0.05）. The Western blotting results showed that compared with blank group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins，and the ratios of phosphorylated MAPK（p-MAPK）/MAPK and phosphorylated ERK（p-ERK）/ERK in the cells in neural differentiation inducer group were increased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were decreased （P<0.05）； compared with neural differentiation inducer group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in Ghrelin group were increased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were decreased （P<0.05）； the expression levels of NSE， NeuN， Tuj-1， NF， and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in U0126 and D-Lys3-GHRP-6 groups were decreased （P<0.05），and the expression levels of Flk1 and CD29 proteins were increased （P<0.05）； compared with Ghrelin group， the expression levels of NSE， NeuN， Tuj-1， NF，and GHSR proteins， and the ratios of p-MAPK/MAPK and p-ERK/ERK in the cells in U0126 group， D-Lys3-GHRP-6 group， and Ghrelin+U0126 group were decreased （P<0.05）， while the expression levels of Flk1 and CD29 proteins were increased （P<0.05）. Conclusion Ghrelin can induce the differention of the ASCs into the neurons，and it mechanism is related to activating the MAPK/ERK pathway.

Abstract:Objective To study the effects of chloroquine on proliferation and cell cycle of hepatocellular carcinoma cell line HepG2 cultured in vitro　and provide basis for research on chloroquine in therapy for liver cancer.Methods HepG2 cells were divided into six groups：control group and chloroquine (8.00，16.00，32.00，64.00 and 128.00 mmol•L^-1 )groups.The cell viability was determined by MTT,the cell cycle and apoptosis were detected by flow cytometry.Results Compared with control group，the cell viabilities were inhibited significantly 24 h after treated with chloroquine (32.00-128.00 mmol•L^-1) or 48-72 h after treated with chloroquine (8.00-128.00 mmol•L^-1) 　( P< 0.01) and the most obvious inhibitory effect occurred when HepG2 cells were treated with 128.00 mmol•L^-1　 chloroquine for 72 h(P< 0.01); HepG2 cells treated with chloroquine (32.00-128.00 mmol•L^-1) for 24 h were obviously arrested at G₂/M phase,compared with control group the percentage of cells at G₂/M phase and the apoptotic rate increased with the dose of chloroquine(P<0.01). Conclusion Chloroquine can suppress the activity of hepatoma HepG2 cells by inhibiting cell proliferation,inducing apoptosis and arresting cell cycle in vitro.So chloroquine may be used as the drug for treatment of hepatocellular carcinoma.

To study the anti-HBV activity and toxicity of a novel MCC-478 devivative 030705 with specific structure in vitro. Methods Anti-HBV activities in HepG2.2.15 cells were analyzed by Southern blotting in experimental groups with different doses of 030705 （0.01, 0.03, 0.10, 0.30 and 1.0 μmol•L^-1）and Adefovir Dipioxil positive control groups(0.10, 0.30, 1.0, 3.0 and 10.0 μmol•L^-1). Concentration of 50% inhibitation (IC₅₀)and concentration of 90% inhibitation (IC₉₀) were obtained. The inhibitory effect of HBeAg was analyzed in HepG2.2.15 cells by ELISA method. The cytotoxicities were tested by MTT method in the experimental groups（10, 30, 100, 300 and 1 000 μmol•L^-1） in HepG2 cells. Concentration of 50% extinction (CC₅₀) was obtained. The inhibitory effects of mitochondrial DNA contents in experimental groups（0.10, 1.0 and 10 μmol•L^-1）were tested by Dot blotting in HepG2 cells. While didenoxycytidine (ddC) groups were as positive control groups and untreated groups were as negative groups. Results Compared with Adefovir Dipioxil positive control groups, the inhibitory effects of the test compound 030705 on HBV DNA had no obvious difference（P>0.05）. Anti-HBV activity of the test compound 030705 was similar to that of Adefovir Dipioxil in HepG2. 2.15 cells. The inhibitory rates of HBeAg in experimental groups （0.01, 0.03, 0.10, 0.30 and 1.0 μmol•L^-1 030705）were 5.94%, 6.08%, 6.32%, 10.31% and 12.49%, respectively in HepG2.2.15 cells; compared with negative control group, there was no obvious difference（P>0.05）. CC₅₀ value of the test compound was 2 014 μmol•L^-1 (>1 000 μmol•L^-1) in HepG2 cells. The inhibitory rates of mitochondrial DNA content from positive drug ddC （0.10, 1.0 and 10 μmol•L^-1）were 38.43%,46.51%,56.51%, respectively in HepG2 cells; compared with negative control group, there was obvious difference（P＜0.05）. The inhibitory rates of mitochondrial DNA content from the test compound 030705 with the same concentrations were 7.00%,5.81%,5.78%, respectively in HepG2 cells; compared with negative control group, there was no obvious difference（P＞0.05）. Conclusion Anti-HBV activity of the test compound 030705 was similar to that of Adefovir Dipioxil in HepG2.2.15 cells. CC₅₀ value of the test compound is 2014 μmol•L^-1 (>1 000 μmol•L^-1) in HepG2 cells. No apparent reductions of mitochondrial DNA content are observed in HepG2 cells. The test compound 030705 with specific structure may be a new promising anti-HBV agent.

Objective To discovery the stress distribution in bone around the implant of middle implant-teeth fixed bridge under different loading. Methods The model was set up by 3-D finite element means. The stress in bone around the implant under loading was computed by finite element analysis software Super-SAP 93. Results The biggest stress in bone around the implant lied in the neck cortex of bone. The biggest stress in bone around the implant under concentrated oblique loading was 2.5 times as big as that under concentrated vertical loading. The peak value of tensile stress lied in the boundary of lingual cortex of bone under concentrated vertical loading. The biggest compressive stress was higher than the biggest tensile stress under concentrated loading. Conclusion It is important not only to absent abnormally high occlusal points, but also to reduce side direction force for arrangement of natural teeth, middle implant, and fixed bridge into balanced occlusion.

Objective To select materials with high quality and safety and evaluate the clinical value of multi-point forming technique in skull repairing. Methods 161 patients suffered from skull defect had been cured in our hospital, within them 43 patients were treated with multi-point forming technique in titanium mesh shaping, 19 patients with traditional handwork shaping, 99 patients with bone-like concrete (acrylate). The following aspects were analyzed and compared：neurosurgeons′ shaping workload before operations,operative time, approving scale on shaping and complications after operation. Results Repairing skull by titanium mesh with the technique of multi-point shaping significantly shortened the average operative time to （30±6） min, compared with acrylate group (70±18 min) and titanium mesh trad itional handwork shaping group (50±11 min) (P<0.01); and decreased the neurosurgeons′　working intensity of shaping time to （5±1 min）, compared with acrylate group (30±5 min) and traditional handwork shaping group (20±3 min) (P<0.05); and improved the patients′ approving scale to 100%, compared with acrylate group (91%) and traditional handwork shaping group (95%) (P<0.05).Conclusion The quality of titanium mesh is obviously better tha n acrylate, the effect of repairing skull with multi-point forming technique is superior to handwork shaping.

Objective To study the effects of whole-body irradiation（WBI）with different doses of X-rays on apoptosis in mouse splenocytes and peritoneal macrophages. Methods The apoptosis percentages of mouse splenocytes and peritoneal macrophaes were detected with flow cytometry（FCM）at different time after the whole-body X-irradiation using the staining of Annexin-V and PI. Results As com pared with the control，the percentage of apoptosis in mouse splenocytes began to increase gradually 24 h after WBI with 2 Gy X-rays（Ｐ<0.05），and the percentage of apoptosis in mouse peritoneal macrophages aros e 4 h after 2 Gy irradiation,and kept higher level till 48 h（Ｐ<0.05，Ｐ<0.01，Ｐ<0.001）. After WBI with 0.075 Gy X-rays，the percentage of apoptosis in mouse splenocytes reduced at 24 h （Ｐ<0.05），and the percentage of apoptosis in peritoneal macrophages significantly reduced from 2 h to 16 h（Ｐ<0.05）as compared with the control. Conclusion High dose irradiation can stimulate apoptos is in mouse splenocytes and peritoneal macrophages， while low dose irradiation can cause an opposite effect and even suppress immunocyte apoptosis.

Objective To study the micrometastasis in lymph nodes of the patients with gastric cancer and its clinical significances. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect CK18 mRNA expression in 298 lymph nodes from 35 patients with gastric cancer and 20 lymph nodes from control group. Results There was no CK18 mRNA expression in control group. In experimental group, the positive rates of CK18 mRNA expression detected with routine histology and RT-PCR were 33.2% (99/298) and 44.6% (133/298),respectively,there was significant difference (P<0.05). Of lymph nodes in which routine histology failed to find metaslasis, 17.1% lymph nodes were found the micrometastasis by CK18 mRNA RT-PCR. Conclusion CK18 mRNA RT-PCR is more sensitive than routine histology for detecting the micrometastasis in lymph nodes of gastric cancer, and has important clinical significances in evaluating clinical staging, predicting prognosis and directing postoperative treatment.

Abstract:Objective To study the effect of Shuganjieyuyin (SGJYY) on monoamine transmitters and their metabolites of brain tissue in depressive mice.Methods The lonely feeding and chronic moderate intensive unpredictable stimulation were used to build depression model and SGJYY was administered. HPLC was used to test the contents of monoamine transmitters and their metabolites of brain tissue in depressive mice and observe the changes of weight and consumption of sugar water and behaivor indexes in mice.Results Compared with normal group,the weight increased slowly(P<0.01), the score of Open-field and sugar water consumption decreased(P<0.01)，the time of no-moving increased(P<0.01) in model group.Compared with normal group,the contents of 5-HT,NE,DA and DOPAC of brain tissue decreased(P<0.01),5-HIAA and MHPG increased(P<0.01) in model group.The 5-HT contents in BYJ and large dosage SGJYY groups were near normal level,the 5-HIAA content in middle dosage SGJYY group was near normal level.The contents of NE in BYY and large dosage SGJYY groups were near normal level,the contents of MHPG in BYJ and anti-depression of SGJYY groups were near normal level.The contents of DA in BYJ and large SGJYY group were near normal level,the DOPAC content in BYJ group was near normal level.Conclusion The mechanism of anti-depression of SGJYY is related to regulating the contents of monoamine transmitters and their metabolites.