Abstract:Objective To investigate the effects of gliadin on development and function of central nervous system in mice,and elucidate that gliadin is very likely to be an environmental factor for mental disorder.MethodsNewly-weaning BALB/c mice were intragastrically administered with gliadin,and stimulated with stress.The mice were divided into positive anti-gliadin IgA aitibody group and negative anti-gliadin IgA antibody group(n=10).The systemic-toxic effects of animal models induced by gliadin were detected with basic observation,ethological evaluation (Hang test,Balance and coordination test,Open-field test and  Forced swimming test) and the cerebral index was calculated.Results The mice in positive anti-gliadin IgA antibody group had less activity and depression,and the body weight  was lower than that of the mice in negative anti-gliadin IgA antibody group (P<0.05).The results of Balance and coordination test,Open-field test and Forced swimming test showed that the mental status was affected more seriously in positive anti-gliadin IgA antibody  group than that in negative group (P<0.05).The ratio of the cerebellum to whole brain in positive anti-gliadin IgA antibody  group was significantly decreased as compared with negative group (P<0.05).Conclusion The immunological reaction induced by gliadin maybe influence the development and function of central nervous system in mice.

Objective To explore the apoptosis inducing effect of vincristine on human osteosarcoma MG63 cells and clarify the molecular mechanisms.Methods The  apoptosis and intracellular reactive oxygen species(ROS) level of MG63 cells were determined by flow cytometry,after incubation with Annexin V-PI and DCFH-DA which were explored on different concerntrations of vincristine (0,5,10,20,50,100 μg/L).Real-time quantitative RT-PCR was used to analyze gene expression.Results 10 μg/L vincristine could significantly increase the apoptotic rate of MG63 cells to 17.6%(P<0.05),the apoptotic rate was increased with the increasing of concentration of vincristine in a dose-dependent manner;5 μg/L vincristine increased the intracellular ROS level and decreased the expression of CAT gene (P<0.05).Conclusion Vincristine can promote the apoptosis of MG63 cells with low concentration,its mechanism is to down-regulate the expresion of CAT gene and induce the increase of ROS to promote apoptosis

Objective
To observe the neuoprotective effect of  lentiviral vector expressing and secreting neuroprotective peptide (NAP) on closed head injury in mice via nose-brain pathway and provide theoretical basis for  treatment of  neurodegeneration diseases by lentiviral vector secreting NAP. Methods 54 adult male  Kunming mice were divided into four groups：control group(n=10),closed head jnjury group (CHI,n=20),rLent/NT4-NAP group(rLent,n=20), and rLent/GFP group  (GFP,n=4).The  mortality,NSS and brain edema in various groups were observed.The pathological changes in brain injury region of various  groups stained by HE were observed.The expressions of GFP in nasal epithelium olfactory nerve and brain tissue in GF
P group were detected under laser confocal microscope.Results The  mortality of mice in rLent group (15%) was significantly lower than that in CHI group（55%）. The NSS in  rLent group  were lower than those in CHI groups at 1,3,5 and 7 d after brain injury（P<0.01）.The percentage of H2O in rLent group  was  lower than that in CHI group at 24 h after brain injury（P<0.01）.HE staining result showed that the  pathological changes in rLent group were  fewer  than those in  CHI group.There was significant GFP in the nasal epithelium,but no fluorescenc
e in olfactory nerve and brain tissue.Conclusion The recombinant rLent/NT4-NAP could efficietnly infect nasal epithelium in vivo.NAP provides significant neuroprotective effect from the complex array of injuries elicited by head trauma.The recombinant rLent/GFP could only infect nasal epithelium,but not go through blood-brain barrier (BBB) to central nervous system (CNS).

Objective
To explore the combined effect of methylprednisolone (MP),electro-acupuncture and  amniotic epithelial cells  (AECs)  transplantation  on axon-plasma transporting function in rats with spinal cord injury(SCI),and provide a more effective method to clinical therapy for SCI.Methods Sixty female Wistar rats were randomly divided into  to 5 groups (n=12).SCI control group: no treatment after SCI was created.MP group: MP pulse treatment after SCI,for 3 d.MP and electro-acupuncture group: electro-acupuncture treatment on Hua Tuo Jiaji acupoint 4 h after SCI,based on MP group.MP,Electro-acupuncture and AECs group: rat AECs transplantation treatment in SCI region 7 d after SCI based on MP and electro-acupuncture group.Sham group: neural scute was opened and spinal cord was exposed without SCI.Ethological observation (BBB scores) was performed every 6 d.Fluorescein red (FR) anterograde tagging and GFAP immunofluorescence histochemistry observation were performed on the 30th day after operation.Results BBB scores revealed that functional recovery diversity of the hind limb was seen in every therapy group after SCI,and the MP,electro-acupuncture and AECs group was the best and the score was 15.23±1.01 on the 30th day.As revealed by FR anterograde tagging,MP,electro-acupuncture and AECs group contained numerous ordinary FR-positive axons,and neural labelled compound could be transported to the far distant area of  SCI region distal end,the number of FR-positive axons of  SCI region were 312.67±34.06 under fluorescence microscope（×100）,the differences between MP,electro-acupuncture and AECs group and other therapy groups were statistically significant（P＜0.01）.GFAP revealed that the quantity of GFAP positive astrocytes in each injured group was increased obviously than sham operation group and MP,electro-acupuncture and AECs group listed the least,the number of GFAP positive astrocytes of  SCI region was 633.61±54.4 under  fluorescence microscope（×200）,there were significant differences between MP,electro-acupuncture and AECs group and other therapy groups（P＜0.01）.Conclusion A combination of methylprednisolone,electro-acupuncture and AECs transplantation can restrain hyperplasy of horizontal cells,promote the recovery of axon-plasma transporting function and the regeneration of axons in SCI rats.

To study the therapeutic effect of topically applied keratinocyte growth factor-2 (KGF-2) in vivo on corneal epithelial wound in rabbit model of alkali burn and explore new methods of treatment in the early of corneal epithelial wound. Methods A 6 mm diameter epithelial defect was made in the center of rabbit corneas by the alkali-soaked filter paper.A total of 60 Japanese white rabbits were randomly divided into two groups(n=30): KGF-2 group (25  mg/L) and control group. A 50 μL aliquot of 25 mg/L recombinant human KGF-2 was then applied topically three times a day. The same volume of phosphate-buffered saline (PBS) vehicle was applied to the control.Each corneal epithelial defect was subsequently photographed with a slit lamp on day 7 and 14 after burn and was measured by a computer-assisted digitizer.Corneal neovascularization in two groups was observed every day.The viability of corneal  epithelial cells   was assessed by MTT assay;meanwhile,the corneal epithelium was examined by morphometric and histological analysis.Results There were statistically significant differences in the epithelial defet area on day 7 and 14 between KGF-2 and control groups(p<0.01).The differences in corneal neovascularization (CNV) between KGF-2 and control  groups were also evident on day 7 and 14, the average percentages of CNV in KGF-2- group were 7.32% and 32.50%,whereas the rates in control group were 17.50% and 41.67%,respectively.On day 7 and 14 after wounding,there were statistically significant differences in the mean 570 nm absorbances between KGF-2 and control groups(P<0.05 or P<0.01).Histological examination showed that the corneal lesions consisted of stromal edema,keratocyte loss,inflammatory cell infiltration,and CNV.Corneal epithelium cell desquamation,stromal edema and numerous neutrophil infiltrations were present in the corneal stroma in control group.In contrast,the KGF-2 group exhibited no corneal stroma inflammation,only mild stromal edema remained in KGF-2 corneas on day 14 and the significant changes of the re-epithelialization in KGF-2 group were observed.Conclusion  Topically applied 25 mg/L KGF-2 can accelerate corneal epithelial wound healing,in contrast to the control,and reduce inflammation,stromal edema,and fibrosis.In addition,it also exhibites significant inhibition on CNV.KGF-2 maybe become another important growth factor in the regulation of corneal epithelial wound healing.

Objective To construct the genetic engineering bacteria highly expressing  23 amino acids human keratinocyte growth factor -1 (rhKGF1_dest23) missing N terminal,and provide experimental data for development of new drug for  treatment of oral mucositis after radiotherapy and chemotherapy.Methods PCR was used to synthese 23 amino acids rhKGF1_dest23 missing N terminal and sumo gene fragments,and construct four kinds of recombinant prokaryotic expression vectors:  pET22b-rhKGF1_dest23,pET22b-sumo-rhKGF1_dest23,pET3c-rhKGF1_dest23 and pET3c-sumo-rhKGF1_dest23,then they were transformed into prokaryotic expression host bacteria: Rosetta(DE3) plysS,BL21(DE3),BL21(DE3)Star plysS,origima(DE3) and BL21AI,the best expression combination of plasmid and host strain of rhKGF1_dest23 protein was screened and purified by CM ion-exchange and heparin affinity chromatography and identified with Western blotting.Results pET22b-rhKGF1_dest23 plasmid and the BL21AI host bacteria was the best combination of expression,after induced by IPTG and arabinose,the majority of recombinant protein was expressed in soluble form,accounting for about 12% of the total bacterial proteins.Its purity reached to more than 95% of the protein after two steps chromatography,then conformed with Western blotting.Conclusion Human genetic engineering bacteria of KGF1_dest23 is successfully constructed and induced by IPTG and arabinose,then after CM weak cation exchange and heparin affinity chromatography,the purified rhKGF1_dest23 protein is obtained

Objective To investigate the protective effects of Flavone mixture of crataegus leave (FMCL) on the rat myocardial ischemia reperfusion models and their mechanisms.Methods Fifty-six Wistar rats were randomly divided into control group,sham group,ischemia reperfusion model group,Xue-Sai-Tong(20 mg/kg) group and FMCL (30.0，15.0，7.5  mg/kg) groups(n=8).The myocardial ischemia reperfusion model was induced by 30 min left anterior descending coronary occulusion and 120 min reperfusion in openchest anesthetized rats to study the effects of FMCL on elegated ST-T of electrocardiogram ( ECG),the size of myocardial infarction,the morphological changes of cardiocytes and the contents of SOD,MDA,CK,AST,LDH and NO in serum were detected. Results Compared with model group,elegated ST-T of  ECG in FMCL（30.0,15.0 mg/kg）and Xue-Sai-Tong groups were decreased（P<0.01）,the sizes of myocardial infarction in FMCL（30.0,15.0,7.5 mg/kg）　and Xue-Sai-Tong groups were shrinked（P<0.05 or P<0.01）,FMCL could ameliorate the pathological lesion. Compared with model group,the contents of AST,CK,LDH and MDA in FMCL（30.0,15.0 mg/kg）and Xue-Sai-Tong groups were decreased（P<0.05 or P<0.01）,and the contents of SOD and NO were increased（P<0.01）.Conclusion FMCL  has the protective effects on myocardial ischemia reperfusion injury in rats,which may be related to inhibition of lipid oxidation.

Objective To study the inhibitory effect of recombinant plasmid pSilencer1.0-U6-siRNA-STAT3  on ovarian cancer in nude mice and clarify the the role of RNA interference(RNAi) technique in the field of biological therapy of ovarian cancer.Methods The subcutaneous transplantated tumor models in 15 nude mice were established with  cell line SKOV3 of human ovarian cancer. The model mice were randomly divided into 3 groups: control group(physiologic saline)，control group of vacancy plasmid,and  recombinant plasmid group (pSilencer1.0-U6-siRNA-STAT3).The recombinant plasmid was transferred into transplanted tumor in nude mice by electron piercing instrument.The growth status of subcutaneous transplanted tumor in nude mice after injection was observed.The protein expression of STAT3 was detected by Western blotting.The morphological changes and apoptosis of tumor cells were observed by HE and TUNEL staining.Results The growth of transplanted tumor had an obvious suppression after transferred.Compared with two control groups,the growth velocity of transplanted tumor in  recombinant plasmid group was decreased(P<0.01),the protein levels of Stat3,CyclinD1,VEGF,Survivin,c-myc in recombinant plasmid group were reduced(P<0.01).HE staining showed the tumor cells in recombinant plasmid group had nuclei pyknosis,fragmentation and other signs of apoptosis.The TUNEL test results confirmed that the tumor tissues in recombinant plasmid group had a large amount of apoptotic cells.Conclusion The recombinant plasmid pSilencer1.0-U6-siRNA-STAT3 can obviously inhibit the growth of ovarian cancer in nude mice.

Abstract:Objective To express the human fibroblast growth factor-21 (hFGF21) in Picha pastoris (P.pastoris) GS115 and SMD1168,and futher provide a basis for treatment of diabetes.Methods According to P.pastoris biased codon and the sequence of hFGF21,the gene full-length coding sequence,which gained by seven rounds of PCR from fourteen 55-59nt oligoes,was cloned into pPIC9K to obtain a recombinant vector pPIC9K-FGF21.And the plasmid pPIC9K-FGF21 was transformed into P.pastoris GS115 and SMD168 by electroporation.The recombinant P.pastoris strains ( His⁺ Mut⁺⁾were obtained by means of MD and MM plates,and high-copy transformants were further screened by increasing G418 concentration and identified by PCR.The positive transfomants were induced by methanol and the highest expressive strain was screend under the optimal conditions of pH6.5 and 0.8% methanol.The expression product was purified by the protocols including ammonium sulfate precipitation,Sephadex G-50 gel filtration,DEAE-Sepharose ion-exchange chromatography. Results The specific fragment of 569 bp was amplified by PCR,sequence analysis was   correct.The results of  SDS-PAGE and Western blotting demonstrated that the culture supernatant of GS115 strains and the negative control strains had no specific band,on the contrary,a specific molecular weight band at about 20 000  appeared in the culture supernatant of SMD1168 strains.Western blotting analysis also showed that the expressed protein could specifically react with anti-hFGF21 antibody,and the hFGF21 was successfully secreted in SMD1168. Conclusion The gene FGF21 has been cloned successfully,it is  expressed and purified in P.pastoris SMD1168.

Objective  To explore the influence of Kangganxian Chongji on the expression of the hepatic collagenⅠ(Col Ⅰ) in the mice with hepatic fibrosis and clarify its theraputical function. Methods 75 female Balb/c mice  were randomly divided into 5 groups (n=15):normal control group,hepatic fibrosis model group,Colchicine group, low dosage therapeutic group and high dosage therapeutic group of Kangganxian Chongji. The hepatic fibrosis model mice were induced by intraperitoneal injection of pig serum respectively (0.5 mL/once,once a week,total 7 weeks) except for normal control group. At the same time the mice in therapeutic group were treated with Colchicine or Kangganxian Chongji once a day,lasted for 7 weeks.The mice were killed and the livers were took from the mice at the end of the 7 th week. The liver tissue slices were made and dyed with HE,and then the pathological changes of the mouse liver were observed  under microscope.The mRNA expression of Col Ⅰa2 and protein expression of  Col Ⅰa1 in the liver tissues were detected with real time PCR and Western blotting respectively.Results The pathological result indicated that the inflammation of liver tissue in Kangganxian Chongji group was alleviated  compared with model group.In low dosage Kangganxian Chongji group,the expression of Col Ⅰα2　 mRNA in liver tissue was markedly reduced compared with model group (P<0.01);at the same time,the expression of ColⅠα1 protein in liver tissue was markedly reduced according to Western blotting analysis compared with model group.Conclusion Kangganxian Chongji can obviously decrease the expression of ColⅠ protein of the liver tissue in the mice with hepatic fibrosis,and the results suggest that Kangganxian Chongji has obvious therapeutic function on hepatic fibrosis.

Abstract:Objective By observing the expressions of vascular endothelial growth factor (VEGF),metallothionein(MT),and matrix metalloprteinase-9 （MMP-9） in the  kidneys of normal and diabetic rats,the influences of exogenous Zn    in kindey and the expessions of VEGF,MT and MMP-9 in kidney tissues of diabetic rats were explored. Methods   Forty-eight Wistar rats were randomly divided into four groups(n=12) :   normal control  (NC)　group,negative control (STZ)group,external Zn intervention diabetic (STZ+Zn) group　and external Zn intervention normal (Zn)　group. The rat diabetic models were made through intraperitoneal injection of streptozotocin(STZ) in STZ and STZ+Zn groups.The rats  in STZ+Zn and Zn groups were orally injected with 5 mg?kg-1of Zn sulphate twice a week. After fed for 16 weeks,the kidneys of  rats were obtained and  organ coefficient of kidney was calculated.The histopathological  changes of  kindeys  and the expressions of MMP-9,VEGF and MT in kidney tissues were detected with immunohistochemistry method. Results Compared with NC group,the organ coefficients  in STZ group and STZ+Zn group were increased (P< 0.05). Compared with STZ group,the organ coefficient in STZ+Zn group was increased(P<0.05). There was no difference of organ coefficient between STZ group and Zn group (P＞0.05).The MMP-9,VEGF, and MT expressed in kidneys of rats.MMP-9 and VEGF mainly expressed in the distal renal tubule;and MT mainly expressed in the proximal renal tubulei. there was no difference  of the expressions of MMP-9,VEGF, and MT in kidney tissues between various groups(P>0.05).Conclusion External Zn intervention can  increase the organ cofficient of kidney in diabetic rats,but dosen’t affect the organ coefficient of kindey in normal rats.  The expressions of  MMP-9,VEGF, and MT  in kidney tissues of  rats have no changes after external Zn intervention.

Abstract:Objective To study the  effects of benserazide on extracellular dopamine level in the striatum of 6-hydroxydopamine- lesioned Parkinson’s disease rat models. Methods The rat models of nigrostriatal denervation of Parkinson's disease were set up by using 6-hydroxydopamine (6-OHDA)，and the control rats were treated with isotonic Na chloride. Then the extracellular dopamine(DA) level in the striatum of sham-lesioned rats treated with benserazide alone and in the striatum of 6-OHDA-lesioned rats treated with benserazide in combination with L-DOPA were detected by using in vivo microdialysis technique. Results In sham-lesioned rats, the extracellular DA level was significantly decreased by  50  mg?kg-1benserazide（P<0.01）.  In 6-OHDA-lesioned rats,the  extracellular DA level showed a marked increase by the combined use of benserazide and L-DOPA, but the time to reach the peak level was significantly prolonged on a dose-dependent manner（P<0.05）. Conclusion Benserazide can reduce the central AADC activity in the striatum of rats with nigrostriatal denervation, resulting in changes in metabolism of exogenous L-DOPA. And  higher dose of benserazide might be a useful agent to prevent adverse effects of long-term L-DOPA therapy.

Abstract:Objective To study the intervention of fluticasone combined with ventolin and montelukast sodium on rat asthmatic airway remodeling and clarify the mechanisms of asthma and airway remodeling. Methods Forty Wistar rats were randomly divided into negative control group, positive control group(model), drug 1 group(inhaled with fluticasone and ventolin) and drug 2 group(administered with montelukast sodium),10 rats in each group. OVA was used to set up rat asthma airway remodeling.The number of inflammatory cells in bronchoalveolar lavage fluid(BALF) was observed, the serum IL-12 levels were detected in various groups, and the changes of ciliated epithelial cells, fibroblasts, and smooth muscle cells were also observed. Results Compared with negative control group, the number of inflammatory cells in rat BALF in positive control group was significantly increased(P<0.05); compared with positive  control group, the number of inflammatory cells in rat BALF in drug 1 group and drug 2 group was significantly decreased (P<0.05); but there was no significant difference between drug 1 group and drug 2 group(P>0.05). Compared with negative control group, the serum IL-12 level of rats in positive control group was decreased(P<0.05); compared with positive control group, the serum IL-12 levels of rats in drug 1 group and drug 2 group were significantly increased (P<0.05 or P<0.01);and there was significant difference between drug 1 group and drug 2 group (P<0.01). Compared with drug 2 group, the airway cilia arranged regularly, the elastic fibers were basically normal, the karyon and cytoplasm had no changes, the smooth muscle mildly thickened, the lymphocytes moderately infiltrated indrug 1 group. Conclusion Fluticasone combined with ventolin can inhibit the infiltration of inflammatory cells, alleviate the airway inflammation, weaken asthma  airway remodeling, and decrease the high airway reactivity to protect the morphology and function of airway.

Abstract:Objective To observe the electroencephalogram (EEG) changes of asphyxial cardiac arrest (ACA) after hypoxic preconditioning (HP) in rats and investigate the activity of EEG in brain ischemic tolerance. Methods Sixty male Wistar rats were randomly assigned to three groups: ①ACA group (n=20),the rats  were subjected to 1 min of ACA and then resuscitated; ② HP+ACA group (n=20),the rats were pretreated with 4 times of 1 min HP 24 h before ACA;③HP group (n=20),the rats  were treated with 4 times of 1 min HP. The mortality, EEG, time from initiation of asphyxia to CA (TCA) and time from resuscitation to restoration of spontaneous circulation (TROSC) were recorded; neuro-functional outcome was assessed by neurodeficit score (NDS). Results The mortality was 5% in HP+ACA group and 30% in ACA group (P<0.01); compared with ACA group, the rats in HP+ACA group showed much shorter TCA and significant longer TROSC（P<0.01）. NDS in HP+ACA group was greatly better than in ACA group (P<0.01); the time to EEG silence and restoration of EEG were significantly shorter in HP+ACA group (P<0.05). Conclusion  Hypoxic preconditioning can induce brain ischemic tolerance through EEG activity modulation.

Abstract:Objective To studey the acute,accumulative and genetic toxicities of carboxymethyl chitosan-Ag+（CMCT-Ag+）in mice,and provide safety guarantee for its clinical application. Methods According to the requirements of The Procedures and Methods of Food Safety Evaluation on Toxicology,ICR mice (SPF) were randomly divided into control group(H₂O),CMCT-Ag+ and SCMC (or CTX) groups.Gastric perfusion (GF) was applied once in ICR mice with the dosage of 5 g?kg-1.14 d later,the LD50 of CMTC-Ag+ in mice was measured,and the organic impairments were evaluated by mice status as well as liver,spleen,and thymus indexes.For the other 14 d,GF was applied twice a day,with 4 h interval,in mice with the dosage of 10 g/kg.The maximum tolerance dosage of CMTC-Ag+ was measured.Then,GF was applied once a day with the LD50 dosage of 5 g/kg.20 d later,the fatality of mice was counted,the accumulation coefficient was calculated,and the organic impairments were evaluated by mice status as well as liver,spleen,and thymus indexes.Finally,GF was applied once in mice for two consecutive days with the CMCT-Ag+ concentrations of 1.25,2.50 and 5.00 g?kg-1.6 h later,the number of micronucleus in  polychromatic erythrocytes (PCE) of bone marrow of mice was counted.Results The LD50 of CMCT-Ag+ in mice was over 5 g/kg;the maximum tolerance dosage was 10.00 g/kg and the accumulation coefficient was more than 5,hence,CMCT-Ag+ could not impair the visceral organs of mice.The micronucleus rates  in 1.25,2.50 and 5.00 g/kg CMCT-Ag+ groups were 3.3‰,1.8‰,and 1.3‰,respectively;there was no significant difference between experimental groups and control group (P＞0.05).Conclusion CMTC-Ag+ exhibits neither toxicity nor mutagenic reaction.It can be used in clinic.

Abstract:Objective To investigate the effect of Gusuibu on the rapid tooth movement in dogs treated by resistance reduction and distraction osteogenesis.Methods A total of 8 local hybrid dogs were selected in this study.The second pre-molar extracted in both sides,four posterior regions in each dog were assigned as regular tooth movement (A),rapid tooth movement (B),regular tooth movement combined with Gusuibu injection (C),rapid tooth movement combined with Gusuibu injection (D).The distance of tooth movement,pulp test and radiography examination were performed for each side on the 2nd,3rd,6th week.Results The average distances of tooth movement on each side were 0.81(A),3.71(B),1.38(C),and 3.67(D)mm,respectively.The distance of tooth movement in side C was significantly bigger than that in side A (P<0.05),and there was no significant difference between sides B and D.By means of X-ray examination,no significant apical root resorption was detected in each side.The stability of moving tooth in side D occurred early than side B.Conclusion During the regular tooth movement,Gusuibu could increase the speed of tooth movement.However,during rapid tooth movement,Gusuibu could not increase the speed of tooth movement,but increase the stability of moving teeth.

echocardiography;acute pulmonary embolism;disease models,animalAbstract:Objective To establish acute pulmonary embolism(APE) model in rabbit under echocardiography,and compare with the pathological results,and explore the feasibility of establishment of APE model monitored with echocardiography.Methods APE models were  established in 25 healthy Japanese white rabbits.The rabbit models of APE  were created by right external jugular vena catheter using gelatin sponge monitored with echocardiography.Gelatin sponge emboli,2 mm×2 mm×10 mm each,following with 5 mL physiologic saline were injected separately to right atrium via the right external jugular vein,which could make these emboli embolize pulmonary artery following blood stream.And the pulmonary artery systolic pressure was detected.Then  the lung tissues slices near embolism place were detected by pathology after the model rabbits were dissected.Results Twenty-three rabbit models with APE were successfully established from twenty-five healthy rabbits.However,one rabbit was unexpectedly dead because of anesthesia,another rabbit was dead owing to acute congestive heart failure of cor dextrum by emboli stagnation in cor dextrum.The echocardiogram of rabbits before and after model establishment showed that the  pulmonary artery systolic pressure was significantly increased after APE,the main pulmonary artery,the left pulmonary artery and the right pulmonary artery were passively expanded.The right ventricle was increased and left ventricle was decreased oppositely,interventricular septum expanded toward left ventricle.there was  significant difference compared with pre-embolism(P< 0.01).Gelatin sponge emboli in the pulmonary artery were detected by pathological detection.Conclusion The method to establish APE model monitored with echocardiography is simple and feasible.It could be used as one of methods to establish APE model,animal.

Objective
To investigate the genetic association between TLR4 gene in chromosome 9q33 and obesity and lung function in Han population from North China.Methods Genomic DNA was isolated from the saliva samples.A single nucleotide polymorphism,rs2149356 (TspEⅠsite) present in the TLR4 gene,was detected using PCR-based restriction fragment length polymorphism(PCR-RFLP) analysis among 506 Chinese Han population.The Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness-of-fit χ2 test.The analysis of forced vital capacity (FVC) and the frequencies of alleles and genotypes of TLR4 gene and the relationship between allelic frequencies and obesity and lung function were statistically computed.Results The result of  t test did not show significant difference of FVC between obesity group and control(P>0.05).The  result of goodness-of-fit χ2 test showed that the genotypic distributions of rs2149356 were not deviated from Hardy-Weinberg equilibrium in various groups (P>0.05).The relevance analysis on allelic gene showed that there were no significant differences in the distributions of the frequencies  of allelic A and allelic C between control and obesity groups (P>0.05).The result of χ2 test did not show allelic association and genotypic association for rs2149356 between obesity and FVC (P>0.05).The  result of goodness-of-fit χ2 test showed that the genotypic distributions of rs2149356 were not deviated from Hardy-Weinberg equilibrium in FVC variant groups(P>0.05),too.Conclusion rs2149356 locus may not be associated with obesity and lung function,but the correlation between the other SNPs locus at TLR4 gene and obesity and lung function could not be ruled out.

Objective
To study the dynamic changes of the levels of Th1/Th2 cytokines in  serum of the patients with chronic hepatitis B(CHB) treated with adefovir dipivoxil,　and  explore  the immune response mechanism of adefovir dipivoxil against hepatitis B virus (HBV). Methods 22 CHB patients were enrolled and given adefovir dipivoxil 10 mg?d-1.The cytokine level，HBV-DNA level and liver function were assessed by three-color flow cytometry before treatment  and 12,24 weeks after treatment.Results  After treatment of adefovir dipivoxil,the average levels of the cytokines secreted by Th1 and Th2 were higher than before trentment (P<0.05).Before treatment，the level of TNF secreted by Th1 was significantly higher than those of  IL-4,IL-6,and  IL-10 secreted by Th2 (P<0.05).12 and 24 weeks after treatment,the levels of the cytokines secreted by Th1 were still higher than those of  the cytokines secreted by Th2，but there was no significant difference (P>0.05).12 weeks after treatment,compared with  control group,the levels of cytokines secreted by Th1/Th2 had no significant change; 24 weeks after treatment,the levels of IFN,IL-4 and IL-6 were higher than those in control group (P<0.05).Conclusion Adefovir dipivoxil is effective in inhibiting HBV replication,promoting the immunologic mechanism of the body.Adefovir dipivoxil can break down the immune tolerance and help to recover T cell function.

Objective
Through the study on HBV P gene sequences of RT resistance mutation patterns and drug resistence of telbivudine in treating with the chronic HBV infeceted patients to discuss the main resistance mutation patterns and drug resistence of telbivudine.Methods The recombinant clone sequencing assay was used to detect HBV P gene sequence  in 47 patients who received telbivudine treatment for 1 year,2 years,and 5 years.  Results In the patients received telbivudine treatment for one year,2 cases(4.26%)had common variable sites,P gene region of mutation patterns was rtM204I;for 2 years,10 cases(21.28%) had variable sites,9 cases of mutation model rtM204I, 1 case mutation model rtL180M + rtM204I;for 5 years, 24 cases(51.06%) had common variable sites variation patterns, 16 cases(66.7%) of rtM204I,4 cases(16.7%)of rtL180M + rtM204I. There were also rtL181V + rtM204I,rtM204V,rtM204I + rtN238H, rtL180M + rtM204I + rtD263E each 1 case. Conclusion The main telbivudine resistance mutation models  are rtM204I and rtL180M + rtM204I.The resistance rates of telbivudine treatment for 1,2,5 years are 4.26%,21.28%,and 51.06%,respectively.And the 5-year resistance rate is significantly lower than that of the 4-year resistance rate of lamivudine,but the resistance rate of telbivudine is higher than that of adefovir.

Objective
To explore the CLDN6 expressions in ovarian cancer,liver cancer and meningioma tissues and clinical significances,and clarify its effects in the occurrence and development of cancer.Methods The expressions of CLDN6 in 36 ovarian cancer tissues,30 liver cancer tissues and 29 invasion meningioma tissues were detected by SP immunohistochemical staining,at the same time,26 benign neoplasm of ovarian tissues,19 normal hepatic tissues and 29 non-invasion meningioma tissues were used as control.Results The positive expression rates of CLDN6 in ovarian cancer tissues and  benign neoplasm of ovarian tissues were  69.44% and 34.62%,there was significant difference between them（P<0.05）；the expression positive rates of CLDN6   in liver cancer tissues and normal hepatic tissues were 73.33% and 26.31%,there was significant difference between then(P<0.05);the positive expression rates of CLDN6 in  invasion meningioma tissues and  non-invasion meningioma tissues were  10.34% and 65.51%，there was also significant difference（P<0.05）.Conclusion The expression of CLDN6 is up-regulated in ovarian cancer and liver cancer tissues,and down-regulated in invasion meningioma tissues. It may have positive or negative regulatory effect on different cancers by destroying compose of tight junctions.

Objective
To detect the changes of FOXD1 expression in invasive breast cancer tissues and tissues surrounding breast cancer,and provide basis for further study on the functions of FOXD1 in the occurrence and development of breast cancer. Methods 113 samples were classified into invasive breast cancer tissue and tissues surrounding breast cancer by frozen section and HE staining.Then the expression levels in four samples of invasive breast cancer tissues and another four samples of tissues surrounding breast cancer were examined by qRT-PCR and Western blotting respectively.Results 113 samples were classified into 36 cases of invasive breast cancer tissues and 77 cases of tissues surrounding breast cancer according to standards of pathohistology.The qRT-PCR result showed that the FOXD1 expression in invasive breast cancer tissues was increased significantly compared with tissues surrounding breast cancer (P<0.01).The result of Western blotting also demonstrated that the FOXD1 expression in invasive breast cancer tissues was higher than that in tissues surrounding breast cancer (P<0.05).Conclusion The expression of FOXD1 in breast cancer tissue is significantly higher than that in  tissues surrounding breast cancer. The result indicates that the expression level of FOXD1 is related to the occurrence and development of breast cancer.

To investigate the expressions of Stat3 and Stat5  in differentiated thyroid carcinoma(DTC) tissues  and their relationships with clininopathological factors,and study the roles of Stat3 and Stat5 in the occurrence and development of DTC.Methods The expressions of Stat3,Stat5,P53,P21 in  42  patients with DTC  including 38 papillary thyroid carcinoma(PTC) and 4 follicular thyroid carcinoma(FTC),30 cases of thyroid adenomas（TA） and 15 normal thyroid tissues (NTT) adjacent to adenomas   were detected by immunohistochemical (SABC) staining method.The positive expression rates in different tissues were compared,and their corelations with clinicopathological factors of DTC   were analyzed.Results The positive expression rates of Stat3 and Stat5 in DTC were significantly increased compared with TA and NTT respectively (P<0.05),their expression rates were closely correlated with pathological typing of DTC(r=0.76，P<0.05;r=0.80，P<0.05);the expression rates of Stats and Stat5 in PTC were lower than those in FTC(P<0.05).The expression rate of Stat5 was significantly higher than that of Stat3 and they had positive correlation(r=0.78，P<0.05).The positive expression rate of P53 in DTC tissue （30.9%） was higher than those in TA(0%) and NTT(0%)(P<0.05).The positive expression rate of P21 protein in DTC tissue was significantly decreased. The expression rates of  Stat3（ 54.8%） and Stat5 （66.7%）were significantly higher than that of P53(30.9%)and there were positive correlations  with  P53 (r=0.77,P<0.05；r=0.72,P<0.05)and negative correlation with  P21（r=－0.87,P<0.05；r=－0.67,P<0.05）.Conclusion The  abnormal expressions of Stat3 and Stat5 may play  important roles in the carcinogenesis of thyroid carcinoma.Perhaps the abnormal expressions of Stat3 and Stat5 may be caused by mutation of p53 and deactivation of p21.The abnormal increase of the expression rates of  Stat5 and Stat3   may lead to the occurrence and development of DTC.

Objective
To investigate the expressions of macrophage migration inhibitory factor (MIF) and vascular endothelial growth factor (VEGF) in ovarian cancer tissues and detect microvascular density (MVD) and elucidate the correlations between MIF,VEGF and the occurrence of ovarian cancer and explore their roles in tumor angiogenesis.Methods The expressions of MIF,VEGF and factor Ⅷ related antigen in normal ovarian tissues and ovarian cancer tissues were detected by immunohistochemical staining.The results were analyzed by Image Pro Plus（IPP）6.0 software.Results There were no visible expressions of MIF and VEGF in normal ovarian tissues,but positive expressions in ovarian cancer.The expressions of MIF and VEGF were increased significantly with the decreasing grade of the ovarian cancer.There were significant differences of the IOD values between well-differentiated,moderately differentiated and poorly differentiated ovarian cancer（P<0.05 or P<0.01）.The MVD in poorly differentiated ovarian cancer tissues (50.83±11.59) was significantly higher than those in normal ovarian tissues(18.16±2.26),well differentiated(19.88±3.92),moderately differentiated(31.1±3.08)  ovarian cancer tissues(all P<0.01).There was positive correlation between the expressions of MIF and VEGF in ovarian cancer tissues (r=0.579　7，P<0.01).Moreover,the expression of MIF was positively correlated with MVD in ovarian cancer tissues (r=0.962　3，P<0.01).Conclusion MIF plays an important role in the oncogenesis and development of ovarian cancer,and it may promote the angiogenesis of ovarian cancer tissues by up-regulating the expression of VEGF.

Objective
To investigate the effect of Compound Shencao Granules(CSCG) on biochemistry indicators in serum and the pathologic changes of liver tissue in rats suffered from alcoholic fatty liver.Methods Sixty rats were divided into six groups (n=10):control group,model group,Zhikeqing Capsule (ZKQC) group,CSCG(high,middle and low doses) groups.The rats in control group were fed with full forage,the rats in  the other groups  were fed with high fat forage plus 380 mL?L-1 of alcohol for 15 d.After successing for the model preparation,the rats in each group were fed on corresponding drugs for 40 d.12 h after the last administration，the blood samples were collected from abdomen-artery and the contents of total cholesterol (TC),triglyeride (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C),alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected,TC/HDL-C,LDL-C/HDL-C and atherogenie index (AI) were calculated.The rats were put to death by cutting through the head,and their anatomized livers were dyed with HE and examined by light microscope.Results Compared with model group,the levels of TC,TG,LDL-C,TC/HDL-C,LDL-C/HDL-C and AI in high dose CSCG group were significantly decreased（P<0.05 or P<0.01),HDL-C was increased (P<0.01) and AI was decreased (P<0.01);under hight microscope, the ingredients of lipid droplet in CSCG groups（high and middle doses）were significantly reduced and the hepatic cells were relieved.Conclusion CSCG has antagonistic effect on alcoholic fatty liver in rats.

Abstract:Objective  To explore  the relationships  between the expressions of Ghrelin,adiponectin,TNF-α in the  peripheral blood of the patients with non-alcoholic fatty liver disease (NAFLD) and occurrence and development of NEFLD and  clinical significances. Methods 50 patients with NAFLD(including 30 cases of simple fatty liver and 20 cases of fatty hepatitis),10 patients with fatty liver and impaired glucose tolerance(IGT),20 cases of fatty liver and dyslipidemia were selected,and 20 healthy subjects were used as control group.The expressions of Ghrelin,adiponectin,insulin and TNF-α in the peripheral blood of the patients and healthy subjects  were detected with ELASI.Results The expressions of Ghrelin and adiponectin in all experimental groups were significantly lower than those in  control group (P<0.01);The expression of TNF-α in all experimental groups were higher than that in control group (P<0.01);adiponectin was negatively correlated with TNF-α,BMI and HOMA- IR(r=－0.811，P<0.01；r=－0.678．P<0.01；r=－0.727．P<0.01)，but  positively correlated with Ghrelin（r= 0.836，P<0.01）. Conclusion Ghrelin and adiponectin play  important roles in the occurrence and development of NAFLD.

Abstract:Objective Through C7 vertebrae channel to treat contralateral C7 brachial plexus root avulsions  to provide a new method to treat brachial plexus root avulsions.Methods 20 corpses were used to compare the lengths of the prespinal and retropharyngeal way and  C7 vertebrae channel way.30 CT scan pictures of adult human neck were selected to measure the maximum angel of cervical vetebrae and lateral maximum angel of cervical vetebrae.36 isolated fresh vertebrae of C5-C7 were chosen as test pieces,and were divided into control group and experimental group which was perforated.　A axial ultimate load compression was performed in all subjects.A comparison was made to figure out the biomechanical difference between the intact and perforated vertebrae.Results The length through vertebrae channel to contralateral inferior trunk was 69.19 mm,25.14 mm shorter than the average length of C7 nerve.T1 and C7 angle of vertical was 9.4°±0.3°.T1 and C7 drag amplitude of vertical was 2.44 mm.T1 and C7 drag amplitude of lateriflection was 0.156 mm.The maximum load of  cervical vetebrae construction test in experimental group was decreased by 14.89% compared with control group,but there was no significant difference(P> 0.05).Conclusion Through C7 vertebrae channel   enough length of brachial plexus is obtained to treat brachial plexus root avulsions of contralateral.

Abstract:Objective To determine the location of the supraorbital foramen(SOF) and infraorbital foramen(IOF) relative to paropia and the medial corner of the eye,and  provide more accurate data for dental and facial surgery.Methods Twenty embalmed adult cadavers (40 sides;16 men,4 women) were dissected to expose the SOFs and IOFs.The location of the SOFs and IOFs was evaluated with direct and photographic measurement.The data gained was analyzed by statistical method.Results The horizontal distances between the IOFs/SOFs and the medial corner of the eye to the distance between the medial corner of the eye and paropia ratios were measured,and their 95% confidence intervals were（0.22，0.31） and （0.34，0.49）.Their linear regression equations were EF=0.58CF+25.02（unit：mm） and  EF=0.51DG+24.20（unit：mm）,respectively.The vertical distances between IOFs/SOFs and the medial/lateral corner of the eye were (25.09±3.36)mm,(23.91±3.31)mm,(25.75±3.34)mm and (26.93±3.88)mm.The horizontal angles between IOFs/SOFs and the medial/lateral corner of the eye were 72.54°±7.13°,66.77°±5.17°,47.45°±6.57°, and 54.69°±8.38°. The vertical angle between the SOFs and the vertical line of pupil was 12.78°±6.82°.Conclusion The horizontal distances between the IOFs/SOFs and the medial corner of the eye and the distance between the medial corner of the eye and paropia have linear correlation;according to the result,the  different data in different subjects  can be calculated.

Abstract:Objective To observe the change of ultrastructure and pathology of desmoplastic small round cell tumor (DSRCT) and recognize the characteristics of DSRCT and improve the standard of diagnosis.Methods One case of primary DSRCT in right leg was observed by light microscope,immunohistochemical method and electron microscope and analyzed with review of the literatures.Results The size of tumor was 3.2 cm×2.4 cm×1.3 cm with gray-yellow on cross-section.Foci of hemorrhage and necrosis were noted.Under light microscope,the tumor was composed of sharply demarcated nests of small rounded or oval cells.The cellular aggregates were surrounded and separated by abundant fibrous connective tissue.The tumor cells were uniform in size and shape,and showed small to moderate amounts of pale cytoplasm with indistinct cell borders.The nuclei were round to oval,with clumped chromatin and marked hyperchromasia.Some cells had one or two indistinct nucleoli.Numerous mitotic figures and areas of necrosis were dentified.The immunohistochemical results showed that the tumor cells were strongly positive for CK,EMA and NSE.There was focal positive staining for desmin with a perinuclear dot-like pattern.However,the tumor cells were negative for CgA,Myogenin,Syn,LCA,SMA,S-100,NF,GFAP,HMB45,HHF-35,CD3,CD10,Actin,CD99, and CD20.Under electron microscope,the tumor cells showed paranuclear cytoplasmic intermediate filaments arranging in globular or whorl array.Conclusion DSRCT occurs both in the abdomen and at other sites.The patients with DSRCT range widely in age.DSRCT has distinctive histopathologic and ultrastructural features.This tumor shows immunohistochemical feature of epithelial,mesenchymal as well as neural multidirectional differentiation.RT-PCR may be served as an important diagnostic adjunct for DSRAT.The prognosis of the patients with DSRCT is very poor.

Abstract:Objective To explore the correlation between the HLA-DRB1 alleles and the chronic hepatitis B of the Han population in Jilin province，and provide a new way for treatment and prevention of chronic hepatitis B．Methods HLA-DRB1 alleles in 398 patients with chronic hepatitis B and 838 healthy controls were analyzed by using the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) technique.Results The frequencies of HLA-DRB1*03，09 alleles in chronic hepatitis B group were higher than those in healthy control group(6.28％ vs  3.58％，P=0.031，OR=1.805； 19.60% vs  12.17%，P=0.001，OR=1.759)．The frequencies of HLA-DRB1*04，10 and 13 alleles in chronic hepatitis B group were lower  than those in healthy control group (6.28％  vs  11.34％，P=0.005，OR=0.524；0.50%  vs  2.39%，P=0.019，OR=0.207；3.27％  vs  5.97％，P=0.044，OR=0.523)．There were no significant differences in the frequencies of the other HLA-DRB1 alleles between   chronic hepatitis B group and healthy control group(P>0.05).Conclusion The HLA-DRB1*03，09 alleles may be susceptible genes for chronic hepatitis B，and HLA-DRB1*04，10，13 alleles may be protecting gene for chronic hepatitis B of Han population in Jilin province．

Abstract:Objective To evaluate the biomechanical effect of rapid maxillary expansion (RME) with different anchorages at the same load conditions in a  three-dimensional finite element model,and provide a theoretical basis for clinical application.Methods Three-dimensional finite element model of maxilla was made.In the model 1,natural teeth were used as the anchorage; in the model 2,both the teeth and the implant were used as the anchorage. Force 20 N in the Horizontal direction was loaded on model 1 and model 2,respectively,and the displacement on the X-axis,Y-axis and Z-axis were calculated.Then the stress distribution on the bone and the teeth were evaluated,and the displacement trend was depicted.Results Both the two models showed that on the cross-section the median palatine suture opened larger at the front,and on the coronal section the median palatine suture opened like a triangle with its base downward.The cown of the tooth shifted buccally while its root shifted  lingually.The displacement in  model 2 showed less than that in  model 1,and the stress in model 2 was also lower than that in  model 1.Conclusion Compared with the natural teeth,using an implant as an anchorage can increase the bone effect while reduce the tooth effect.

Abstract:Objective To analyze the reason of dyeing and inflammation of gum caused by the non- precious metal porcelain crown,make improvements on the methods of gold-plated and provide a reference for the clinical application of gold-plated porcelain crown.Methods 240 patients who needed porcelain-fused-to-metal crown(PFM) repair from June,2005 to December,2006 were selected.The differences of nickel-chromium alloy,gilded nickel-chromium alloy,and aurum alloy  (PFM) were introduced to patientsThe patients were divided into three groups according to their voluntary choice of PFM.Gilded Ni-Cr alloy PFM group: 80 cases of gilded Ni-Cr alloy PFM; Ni-Cr alloy PFM group: 80 cases of Ni-Cr alloy PFM; Aurum alloy PFM group: 80 cases of aurum alloy PFM.Then  the gum discoloration and gingival health in these groups were observed and compared.Results After the first wearing of PFM,three cases of gum discoloration occurred in Ni-Cr alloy group while no cases occurred in other two groups;but there was no significant difference between Ni-Cr alloy PFM and gilded Ni-Cr alloy PFM and aurum alloy PFM groups(P>0.05)；and there was no gingival inflammation in the three groups.One year later,there were twelve cases of gum discoloration in Ni-Cr alloy PFM group,while no cases occurred in gilded Ni-Cr alloy PFM and  aurum alloy PFM groups,there was  significant difference between them(P<0.01).　Thirteen cases of gingival redness and swelling symptoms appeared in Ni-Cr alloy group and three cases occurred in gilded Ni-Cr alloy PFM group;by means of rank sum test,the difference between them was significant (P<0.01);there was no cases occurred in in aurum alloy PFM group;the difference between Ni-Cr alloy group and aurum alloy PFM group was significant,too(P<0.01),but the  difference between gilded Ni-Cr alloy PFM group and aurum alloy PFM group was not significant(P>0.05).Conclusion  In view of the esthetic effects of gingival margin and moderate price,highly gilded Ni-Cr alloy PFM is an ideal choice in clinical rehabilitation.

Abstract:Objective To study the clinical value of cotreatment with rectal wall repair and procedure for prolapse and haemorrhoids（PPH) on outlet obstruction constipation（OOC）induced by rectocele in woman.Methods The woman rectocele patients diagnosed and meeting the surgical indications in our hospital from 2004 to 2008 were randomly divided into monotreatment with rectal wall repair group and cotreatment with rectal wall repair and PPH group,and the efficiencies of the procedures and Longo’s ODS score were observed.Results The total efficiency in monotreatment with rectal wall repair group was 87.71%,the total efficiency in cotreatment with rectal wall repair and PPH group was 97.5%,there was significant difference between two groups (P<0.01); The Longo’s ODS score in monotreatment with rectal wall repair group was decreased from preoperative 17.34±2.77 to postoperative 6.31±1.31,the Longo’s ODS score in cotreatment with rectal wall repair and PPH group was  decreased  from preoperative 17.25±2.81 to postoperative 5.39±1.62. The  life qualities were improved significantly in both groups. But the effect of combined treatment with rectal wall repair and PPH in patients with  OOC caused by rectocele was more effective than that of monotreatment with rectal wall repair (P<0.01).No major postoperative complications occurred.Conclusion Cotreatment with rectal wall repair and PPH seems to be an effective and safe procedure to treat OOC caused by rectocele; and the procedure is more efficient than monotreatment with rectal wall repair.

Abstract:Objective To discuss the application and effectiveness of video-assisted thoracoscopic (VATS) extended thymectomy for myasthenia gravis (MG).  Methods A retrospective review of 34 MG patients who received VATS extended thymectomy from September 2007 to September 2009 (VATS group) was undertaken,and compared with 27 MG patients who received trans-sternal (TS) thymectomy from November 2005 to May 2008(TS group). The operation time,blood loss quantity,analgesic pethidine dose,hospitalization time,complication,incidence of MG crisis   of the two groups were analyzed and the climical improvement was compared.  Results 34 cases of VATS group successfully received thymectomy and anterior mediastinal fat dissection. There were no operative complications and death cases. One case of MG crisis was reported (2.94%). When VATS group was compared with TS group,the blood loss quantity(102.64 mL±65.51 mL vs 187.40 mL±68.08 mL),the probability of postoperative complications (0.00% vs 11.76%),the postoperative analgesic pethidine dose (19.11 mg±30.19 mg vs 62.96 mg±77.94 mg) and the postoperative hospitalization time (9.64 d±5.53 d vs 13.89　d±6.75　d) were decreased,there were significant differences between two groups(P<0.05). The patients in VATS group were followed up 6 to 28 months(mean 12.7 months). Overall clinical improvement at follow-up was observed in 28 of 33 (84.85%) patients,with 9 of 33 (14.0%) patients in complete stable remission(CSR),compared with TS group(84.64%,23.08%) there was no statistically significant difference(P>0.05). Conclusion VATS extended thymectomy is safe and effective,which is more advantageous for MG patients because of less tissue injury,lighter pain,lower incidence of complication, shorter hospitalization time,better cosmetic result and equ
ivalent CSR rate.

Abstract:Objective To discuss the relationship between coagulation function and gastrointestinal hemorrhage in premature infants.Methods The prothrombin time（PT）,part-time activation thromboplastin(APTT）,thrombin time (TT),fibrinogen(Fib),D-dimer(D-D) in  132 hospitalized   premature infants with gastrointestinal hemorrhage (observation group)and 156 non-hemorrhage premature infants (control group) were detected.The difference of coagulation function between two groups was compared.Results Compared with control group,the PT,APTT,and TT in observation group were significantly increased（P<0.01）,Fib was decreased（P<0.01） and  D-D was obviously increased（P<0.01）.Conclusion The changes of coagulation function are closely related to gastrointestinal hemorrhage in premature infants and may be involved in the pathogenesis of gastrointestinal hemorrhage in premature infants.

Objective To assess the agreement of central corneal thickness (CCT) measured by anterior chamber-optical coherence tomography(AC-OCT) and ultrasonic pachymeter and provide an objective basis for clinical application of  AC-OCT. Methods CCT of 150 college student volunteers (300 eyes) measured by two devices were obtained. The data was analyzed by paired t test and Pearson correlation analysis. Bland-Altman plot and Mountain plot were used to assess the agreement. Results The mean CCT values were（530.05±33.611） μm measured by AC-OCT and （543.68±35.088）μm measured by ultrasonic pachymeter. Regression analysis showed a high correlation between the values obtained by both devices (r=0.960, P<0.001). Compared with AC-OCT, ultrasonic pachymeter overestimated the CCT by a mean of 13.62 μm. The two modalities had incomparable results. Conclusion It is important to be noted in clinical practice that the measurements acquired by these two modalities are not directly interchangeable. However, the CCT measurements by the AC-OCT and ultrasonic pachymeter are highly correlated. AC-OCT is an effective method to observe the changes of the corneal thickness in the long term.

Objective Using magnetic resonance imaging(MRI) technique as control standard,to explore the feasibility and accuracy of true real-time three-dimensional volume quantitative techniques (RT-3DE) in evaluation of  the left ventricular volume (LVV) and left ventricular ejection fraction (LVEF) of the maintained hemodialysis (MHD) patients.Methods 48  MHD patients were enrolled in this study.RT-3DE was used to detect the LVV and LVEF,while  Simpson’s method and M-Teichholz method were used at the same time.The results of the three methods were compared with the data measured by MRI.Results ①The data  measured by  RT-3DE method   showed a highly positive correlation  with  the   LVV detected by MRI  (r=0.90,P<0.001),but there was no significant  difference between the two methods (P>0.05).②When heart cavity deformed,the results of left ventricular end-diastolic volume(LVEDV) and left ventricular end-systolic volume (LVESV) measured by M-Teichholz method and Simpson’[KG-*3]s method were higher or lower than the data measured by RT-3DE method,and the differences were statistically significant (all P<0.05).Conclusion RT-3DE can accurately measure the volume of normal cardiac chambers and the left ventricular cavity deformation volume. The result of RT-3DE has higher accuracy compared with Simpson’s method and M-Teichholz method.
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Objective To identify the diagnosis value of ^99mTcO^－₄/^99mTc-MIBI combined with imaging in thyroid carcinoma by comparing the diagnostic results of ^99mTcO^－₄/^99mTc-MIBI combined with imaging and neck color Doppler ultrasonography (CDU) in thyroid carcinoma,postoperative recurrence or cervical part metastasis. Methods 150 patients with thyroid carcinoma treated by operation and 21 patients who were carried out the second operation because of the suspicion of postoperative recurrence or cervical part metastasis were selected. The pathological results were used as the gold standard and  the diagnostic values of ^99mTcO^－₄/^99mTc-MIBI  combined with imaging and neck CDU in analyzing 437 cases of thyroid carcinoma，215 cases of thyroid carcinoma which diameter was greater than 1 cm and 87 cases of suspected postoperative recurrence or cervical part metastasis were compared respectively. Results In the diagnosis of thyroid carcinoma,the sensitivity of CDU(89.10%) was higher than that of ^99mTcO^－₄/^99mTc-MIBI combined with imaging (81.99%);the patho-coincidences of CDU and ^99mTcO^－₄/^99mTc-MIBI combined with imaging were 84.67% and 80.78%, respectively,there was no significant difference（P>0.05).In the diagnosis of thyroid carcinoma which diameter was great
er than 1 cm,the sensitivity of ^99mTcO^－₄/^99mTc-MIBI combined with imaging（97.75%）was higher than that of CDU（89.89%）；the patho-coincidences of CDU and ^99mTcO^－₄/^99mTc-MIBI combined with imaging were 84.65% and 87.44%,respectively,there was no significant difference（P>0.05）.In the diagnosis of suspected postoperative recurrence or cervical part metastasis,the sensitivity and specificity of CDU（44.78%,40.00%）were lower than those of  ^99mTcO^－₄/^99mTc-MIBI combined with imaging（64.18%,80.00%）,the patho-coincidences of CDU and ^99mTcO^－₄/^99mTc-MIBI combined with imaging were 43.68% and 67.82%，respectively,there was significant difference（P<0.05）. Conclusion ^99mTcO^－₄/^99mTc-MIBI combined with imaging is more sensitive in large diameter thyroid carcinoma,especially has obvious advantage in diagnosing postoperative recurrence or cervical part metastasis.

Objective
To assess the  sleep quality of residents in Changchun city,investigate and analyze its relevant influencing factors,and  provide basis data of the sleep quality of  general population in Changchun city and scientific evidence for prevention and improvement of sleep quality.  Methods From September to November in 2008,3 285 residents in Changchun city  were surveyed as subjects with questionnaires through convenient cluster sampling method,including Chaoyang County,Luyuan County,Kuancheng County,Nanguan County,and Nong’an County; Pittsburgh Sleep Quality Index (PSQI) questionnaire was administered to evaluate the sleep quality.Those whose global score of PSQI over 8 were thought to have poor sleep quality and the prevalence was calculated; Self-designed questionnaire was used to investigate the sleep influencing factors,including demographic characteristics,physical and psychological health conditions.Univariate and multivariate analysis were carried out to explore the correlation between sleep quality and its relavant factors.Results The  mean global score of PSQI was 4.43±2.70 and 12.6 percent residents in Changchun city had poor sleep quality.Univariate factor analysis showed that sex,source(City/Town),occupation type,education level,marriage,abdomen fat,smoking habit were related with sleep quality（P<0.05）.Multivariate factor analysis found out that age,coronary heart disease,body pain,fatigue,low self-satisfaction,family conflict and bad mood were risk factors of sleep quality.BMI,drinking alcohol, hypertension,and diabetes had no significant correlation with sleep quality yet.Conclusion Poor sleep quality exists widely in Changchun residents.Female,rural,manual labor,and poor marriage population have high risk and should arouse genereal concern.Health physical and psychological conditions are advocated for good sl
eep quality.

Objective To study the changes of bone mineral density and incidence of osteoporosis in vegetarians.Methods Dual energy X-ray absorptiometry was used to measure the bone mineral densities of spine,neck of femur and greater trochanter in 62 vegetarians(vegetarian group) and  60 normal age-matched men(control group).Results Compared with control group，the bone mineral densities(tms?cm-2) of spine,neck of femur and greater trochanter  in vegetarians were evidently decreased （0.752±0.075  vs  1.014±0.096，0.697±0.071  vs  1.003±0.111，0.713±0.083  vs  1.011±0.097，P<0.001）and the incidences of osteoporosis and osteopenia were increased（40.3%  vs  13.3%，19.3%  vs  5.0%，P<0.001）.Conclusion Vegetarians have  lower bone mineral density and higher incidences of osteoporosis and osteopenia than omnivores.

Abstract:Objective To establish  a TaqMan PCR-based method for the detection of Mycoplasma in cell culture through designing  a pair of primers and Taq- Man  probe  according to conservative sequence of Mycoplasma 16s ribosomal RNA gene.Methods 16srRNA Mycoplasma species-specific region was selected as amplified regions,and the Mycoplasma universal primers were designed.The expected size of PCR amplified fragments was 288 bp.Recovery of PCR amplified products,connection of pMD-18T vector,transformation of  E.coli JM109,coating of Amp+ agar plate were made,and the positive clones were picked for  PCR identification,and then the  plasmids were  extracted.The  fluorescence quantitative PCR absolute quantification of standard was constructed, the fluorescence quantitative PCR primers and probe were designed,matrix method was used to optimize the reaction system,An absolute quantitative standard curve was established.Ten-fold dilution method was used to detect the sensitivity and compared with the general PCR method and culture method.The specificities of  5 kinds of gram-positive bacteria and Mycoplasma were  tested.The sensitivities of the samples were compared between  the culture method,general-PCR and fluorescence quantitative PCR assay.Results The size of PCR product  of Mycoplasma  was about 288 bp,the plasmid concentration was 13.9 mg/L.A260/A280 (Ratio) was 1.780. The plasmid concentration was 4.25 × 109copy/ μL.10 μmol/L primer concentration and 10 μmol/L probe concentration were the best response to the concentration.Two-temperature cycles and 60℃ annealing temperature were the best cycling conditions.The detection sensitivity was 4.25 ×101 copies.The expression of the linear relationship between copy number and Ct value: Ct =－2.916 × log copy number of +40.02.In  specificity  test,the   five kinds of gram-positive bacteria samples were negative,Mycoplasma sample was positive,indicating good specificity. CV of the accuracy of detection was small,indicating good stability.The same concentration samples got the same Ct value,indicating good reproducibility.The positive samples obtained by fluorescence PCR were  higher than those  ordinary PCR and culture method in sample test.Conclusion The detection system based on real-time RT-PCR is rapid,consistently sensitive and steady,which could be used to detect the laboratory samples and clinical samples.

Objective
To establish the high performance liquid chromatography  (HPLC) method for determination of  2,3,5,4′-tetrahydroxy-stilbene-2-O-β-D-glucoside in Taohong Qingxue Capsule,and provide  basis for   the quality control of industrial production.Methods The HPLC method   was developed to determine 2,3,5,4 ′-tetrahydroxy-stilbene-2-O-β-D-glucoside in Taohong Qingxue Capsule. The developed conditions of HPLC were as follows: the shim-pack VP-ODS C18(150.0 mm×4.6 mm,5 μm) was used,the mobile phase was Acetonitrile - water（17∶83）,the detection wavelength was at 322nm,and the column temperature was 30℃.Results The linear range of 2,3,5,4′-tetrahydroxy-stilbene-2-O-β-D-glucoside was 0.284-2.272  μg.The regression equation was: Y=－25 678+1 204 880X，r=0.999 9.The average recovery was 99.4%,and the (RSD) was 2.29% (n=6). The RSD in accuracy,stability and repeatability tests  were 0.46%,2.89%, and 2.39%,respectively.The content of 2,3,5,4′-tetrahydroxy-stilbene-2-O-β-D-glucoside in Taohong Qingxue Capsule was limited not less than 0.8 mg per one capsule under the developed condition. Conclusion This method is accurate，simple and reliable,which provides the basis for  quality control of Taohong Qingxue Capsule.

Objective To explore the fermentation conditions of ergosterol yeast engineering strain and illustrate that  the yeast strain (YDH/pGAD-ScERGI) can synthesize ergosterol by using corn hydrolysates as fermentation substrates.Methods The different substrates,nitrogen sources,corn syrup contents,inorganic salts,pH were used to measured the cell biomass of engineered strain and ergosterol content.Results When the fermentation parameters of corn hydrolysates 10o Bx, corn steep 6.5%, magnesium sulfate 0.1%, initial pH of 6.5,inoculum 10%,28℃ shaking cultured 16 h,the yeast（YDH/pGAD -ScERG1）cell biomass reached 14.34 g?L-1,the  ergosterol content was  up to 3.05%,ergosterol yield was 435.93 mg?L-1. Conclusion The ERG1 gene expression of yeast strains（YDH/pGAD -ScERG1）is conducive to the synthesis of ergosterol.