Objective: To investigate the damage effect of bisphenol A (BPA) on the testis tissue of adult mice, and to reveal the reproductive toxicity of BPA in the body of animal and mechanism.Methods: 40 KM mice aged 8 weeks were randomly divided into control group (according to the weight ratio of corn oil gavage), low dose of BPA group (100 mg·kg^-1 BPA), moderate dose of BPA group (200 mg·kg^-1 BPA), and high dose of BPA group (400 mg·kg^-1 BPA).4 weeks laster, the testis tissue was taken.The apoptotic rates in the testis tissue were detected by flow cytometry; the distribution and expression of Fas and FADD were measured by immunohistochemistry.Results: Compared with control group, the apoptotic rate, the expression rates of Fas and FADD in testis tissue of the mice in low dose of BAP group had no changes (P>0.05), while the apoptotic rates in the testis tissue and the positive expressions rates of Fas and FADD in moderate and high doses of BPA groups were increased (P<0.05). Compared with low dose of BPA group, the apoptotic rates and the positive expression rates of FAS and FADD in tests tissue of the mice in moderate and high doses of BPA groups were significantly increased(P<0.05). Compared with moderate dose of BPA group, the apoptotic rate and the positive expression rates of FAS and FADD in testis tissue of the mice in high dose of BPA group was significantly increased (P<0.05).The overexpression of Fas and FADD was positively correlated to the apoptotic rate in testis tissue in moderate dose of BPA group (r=0.430, P<0.05; r=0.238, P<0.01) and high dose of BPA group (r=0.637, P<0.01; r=0.359, P<0.01).Conclusion: BPA with content dose can increase the apoptotic rates of cells in testis tissue and the expressions of Fas and FADD.BPA's reproductive toxicity may be closely associated with the activation of Fas signal pathway and resulting in massive apoptosis.

Objective: To study the influence of "Warming Yang and Nourishing Qi" Acupuncture in the expression levels of acetylcholine receptor protein(AchR) in gastrocnemius neuromuscular junction (NMJ) of the rats with experimental autoimmune myasthenia gravis(EAMG), and to clarify the mechanism of "Warming Yang and Nourishing Qi" Acupucture in the treatment myasthenia gravis(MG).Methods: The female Lewis rats were used to establish the EAMG rat models immuned with AchRα1 129-145 polypeptide fragments.30 EAMG rats were selected randomly and divided into model control group, drug control group, and acupuncture treatment group(n=10), and another 10 rats purchased at the same period were used as blank control group.The rats in drug control group were treated with 18.5 mg·kg^-1·d^-1 pyridostigmine by gavage.And the rats in acupuncture treatment group were treated with irritating by "Warming Yang and Nourishing Qi" Acupuncture.The rats in blank control group and model control group were not treated.The Lennon symptom scores of the rats in various groups were recorded.After treatment, the tissues of gastrocnemius NMJ of the rats were taken, and the AchR protein expression levels in NMJ of the rats in various groups were detected by Western blotting method.Results: Compared with blank control group, after successfully modelling, the Lennon symptom scores of the rats in model control group, drug control group and acupuncture treatment group before treatment were significantly increased(P<0.01).Compared with model control group, the Lennon symptom scores of the rats in drug control group and acupuncture treatment group after two treatment course were significantly increased(P<0.01).Compared with blank control group, the AchR protein expression level in gastronemus NMJ of the rats in model control group was significantly decreased(P<0.01).Compared with model control group, the AchR protein expression levels in gastronemus NMJ of the rats in acupuncture treatment group and drug control group after treatment were significantly increased(P<0.01).Conclusion: "Warming Yang and Nourishing Qi" Acupuncture has good therapeutic effecton the EAMG through increasing the NMJ AchR protein expression level of the EAMG rats.

Objective: To investigate the stress distribution surrounding an implant under different thickness of cancellous bone and cortical bone, and to analyze the influence of thickness ratio and total thickness of bone tissues on the reliability of an implant.Methods: By using the commercial finite element method software Abaqus, a simplified three-dimensional model of a jawbone consisting of a cancellous bone, a cortical bone, an implant, and a ceramic crown was constructed, and then the computation was performed.Under the condition that the system was loaded by lateral and normal stresses, the influence of thickness ratio and total thickness of cancellous bone and cortical bone in the stress distribution surrounding the implant was studied, where the thickness ratios were 3:1, 2:1, 1:1, 1:2, and 1:3; the total thickness were 0.5, 1.0, 2.0, 3.0 and 4.0 mm, respectively.Results: The maximum stresses on the cortical bone, the cancellous bone as well as the implant were all found to decrease with the increasing of the total thickness of cortical and cancellous bones, with a higher decreasing rate in the range between 0.5-2.0 mm and a lower decreasing rate between 2.0-4.0 mm.More importantly, the maximum value of stress in the cortical bone within the neck region of the implant was observed to increase dramatically via reducing the total thickness below 2 mm, while it was increased insignificantly when the total thickness was above 2.0 mm.Conclusion: The thickness ratio and the total thickness of cancellous bone and cortical bone have strong influence in the stress distribution surrounding the implant. In dental implantation surgery, the total thickness of cancellous bone and cortical bone should be at least 2 mm, and therefore 2 mm is an optimal value.

Objective: To study the construction of matrix metalloproteinase 3(MMP3) biosensor vector, and to illuminate the activated process of MMP3 in the living cells.Methods: The ECFP-MMP3-YPet biosensor vector anchored on cellular surface was constructed and identified.The MMP3 biosensor was transfected into the 293T cells.The transfection efficiency was observed 24h after transfection.The flurorescence resonance energy transfer(FRET)-based MMP3 biosensor was observed by inversion fluorescence microscope.Results: The MMP3 biosensor vector was successfully constructed.The length of MMP3-YPet identified by double enzyme digestion and PCR was about 780bp.The transfection efficiency of MMP3 biosensor was about 40%, and which was evenly presented in cytoplasm of 293T cells.And the FRET ratio of MMP3 biosensor was decreased after stimulation with uPA on the 293T cells.The FRET ratio reached its minimum about 30 min later. Conclusion: The MMP3 biosensor can sensitively and reliably monitor the MMP3 activation in living cells.

Objective: To study the role of 14-3-3εand Cdc25B in germinal vesicle (GV)-stage arrest of mouse oocytes, and to pay foundation for further study on the molecular mechanism of PKA/Cdc25B/14-3-3εpathway in GV-stage arrest of mouse oocytes.Methods: The eukaryotic expression vectors of pcDNA3.1-ZEO-HA-14-3-3ε, pcDNA3.1-MYC-Cdc25B-WT, pcDNA3.1-MYC-Cdc25B-S321A, and pcDNA3.1-MYC-Cdc25B-S321D were transcribed into mRNA in vitro.The mouse GV-stage oocytes were collected after superovulation and divided into no injection group, TE buffer microinjection group, 14-3-3εmRNA injection group, 14-3-3εmRNAs+Cdc25B-WT mRNA injection group, and 14-3-3εmRNA+Cdc25B-S321A mRNA injection group, 14-3-3εmRNA+Cdc25B-S321D mRNA injection group.The protein expression levels of HA-14-3-3ε and MYC-Cdc25B and the phosphorylation status of Cdc2-pTyr15 were observed by Western blotting method.The morphological changes and germinal vesicle breakdown(GVDB) rates of mouse oocytes were observed under phase-contrast microscope.Results: None of the oocytes in no injection group, TE buffer microinjection group, 14-3-3εmRNA injection group, 14-3-3εmRNA+Cdc25B-WT mRNAs injection group and 14-3-3εmRNA+Cdc25B-S321D mRNA were able to undergo GVBD until at least 20 h after injection(P>0.05);the GVBD rates of oocytes in 14-3-3εmRNA+Cdc25B-S321A mRNA group at 1 h(5.00%±0.68%), 2 h (62.00%±3.56%) and 3 h(100.00%±0.00%) after injection were significantly higher than those in no injection group and TE buffer injection group(P<0.01);the oocytes in 14-3-3ε mRNA+Cdc25B-Ser321A mRNA group at 20 h(79.00%±2.80%) after injection progressed to MII(P<0.01).Conclusion: 14-3-3ε can regulate the transition from GV to GVBD of mouse oocytes by means of phosphorylation and dephosphorylation of S321-Cdc25B.

Objective: To investigate the inhibitory effect of 18β-glycyrrhetinic acid (18β-GA) on the occurrence and development of gastric tumor in the K19-C2mE transgenic mice, and to elucidate the inhibition mechanisms of 18β-GA on inflammation and tumorigenesis.Methods: K19-C2mE transgenic mice with gastric tumor were randomly divided into control group (n=36) and experimental group (n=34).The mice in experimental group were given the water with 0.1%18β-GA, while the mice in control group were given normal water.All animals were sacrificed at 52 weeks, the morphology and size of tumors were observed and measured respectively; the pathomorphology of stomach tissue was observed by hematoxylin and eosin (HE) staining; immunohistochemistry method and the H-score were used to detect the expression levels of cyclooxygenase-2 (COX-2) and interleukin-1β (IL-1β) in stomach tissue.Results: The incidence of gastric tumor in experimental group was significantly decreased compared with control group (41.18% vs 77.78%, P=0.002).The morphology of gastric mucosa was regular in experimental group, the size of gastric tumor was smaller, and the surface of tissue was more smooth compared with control group;little hyperemia and ulcer were observed.The response of inflammation was mild in experimental group, and there were little atypia in cellular morphology and organizational structure.Compared with control group, the expression levels of COX-2 and IL-1β in stomach tissue of the mice in experimental group were decreased (P<0.001).Conclusion: 18β-GA could reduce the incidence of gastric tumor in the K19-C2mE transgenic mice.18β-GA may inhibit the occurrence and development of gastric tumor by regulating the expression levels of COX-2 and IL-1β.

Objective: To construct the Escherichia coli (E. coli) expression system for preparation of the bone morphogenetic protein-2(BMP2) with collagen-binding domain(CBD), and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods: CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropyl β-D-thiogalactopyranoside (IPTG) at 37℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22 μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation.The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results: The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21, and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L^-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14000.Two bands (14000 and 28000) were seen in the SDS-PAGE picture, which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg·L^-1, respectively, and the recovery rate was about 73%.Conclusion: The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully, and the recombinant CBD-BMP2 is expressed and refolded efficiently.The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

Objective: To investgate the ease effects of local injection of ursolic acid both on orthodontic tooth movement distance and tooth root resorption in the rats, and to clarify its mechanism.Methods: Ninety-six male Wistar rats were used to establish models of orthodontic tooth movement. After the successful establishment of models, all the model rats were randomly divided into 0 (control group), 0.5, 1.0 and 2.0 mmol·L^-1 ursolic acid groups(n=24).The rats were locally injected with different doses of ursolic acid into the palatal submucosal area adjacent to the right upper first molar, once per 3 d, each time 50 μL.The rats were respectively sacrificed on the days 1, 3, 5, 7, 10, 14, 21 and 28, then the distances of tooth movement were measured.HE staining was used to observe the morphologic changes of the root tissue of rats.Results: The tooth movement distances of rats in control, 0.5, 1.0, and 2.0 mmol·L^-1 ursolic acid groups were increased with the prolongation of time(P<0.01).Compared with control group, the tooth movement distances of rats in 0.5 mmol·L^-1 ursolic acid group 3, 7, 14, 21 and 28 d after forcing had statistical significance(P<0.05 or P<0.01); the tooth movement distances of rats in 1.0 and 2.0 mmol·L^-1 ursolic acid groups 1 d after treatment had no statistical significance(P>0.05), but at the other time points, the tooth movement distances of rats had statistical significance(P<0.01). The tooth movement distances of rats had statistical significance between 0.5, 1.0 and 2.0 mmol·L^-1 ursolic acid groups 5, 7, 10, 14, 21 and 28 d after forcing (P<0.05 or P<0.01). The morphological results showed that the resorption pits were found on the surface of tooth root with the prolongation of loading time;in addition, with the increasing of the doses of ursolic acid, the absorption of root surface was relieved. Conclusion: Local injection of ursolic acid at the doses of 0.5, 1.0 and 2.0 mmol·L^-1 can reduce the distance of orthodontic tooth movement in the rats, and the distance of tooth movement in the rats is reduced with the increasing of its doses, and ursolic acid has the ease effect on the orthodoutic tooth root resorption.

Objective: To study the effects of Hedgehog signal transduction pathway on the cell proliferation, apoptosis and connexin 32(Cx32) and connexin 43(Cx43) membranous distribution of breast cancer cells, and to explore its mechanism in the cell proliferation and metastasis of breast cancer.Methods: The breast cancer MCF-7 cells at logarithmic growth period were divided into cyclopamine groups and blank control groups.The MCF-7 in cyclopamine groups were treated with 5, 10, 20, 30 and 40 μmol·L^-1 for 24, 48 and 72 h;MTT assay was applied to detect the inhibitory rate of proliferation of MCF-7 cells.After the MCF-7 cells were treated with 0(negative control group) and 25 μmol·L^-1 cyclopamine for 48 h, flow cytometry was employed to determine the apoptotic rate and to analyze membranous distribution of Cx32 and Cx43 in the MCF-7 cells.Results: Compared with blank control group, the inhibitory rates of proliferation in cyclopamine groups were increased(P<0.05), and the inhibitory effect of proliferation was increased with the increasing of cyclopamine doses and prolongation of treatment time. After treated with 25 μmol·L^-1 cyclopamine, the apoptotic rate of MCF-7 cells was higher than that in blank control group(P<0.05).The positive expression rates of Cx32 and Cx43 48 h after treatment were higher than those in negative control group(P<0.05).Conclusion: Hedgehog signal transduction pathway can inhibit the apoptosis and mediate membranous distribution of Cx32 and Cx43 in breast cancer cells.

Objective: To explore the influence of rosuvastatin in the lipid levels, atherosclerosis plaque and apoptosis in the plaque of atherosclerotic apolipoprotein E(ApoE)-knockout mouse models, and to clarify the mechanism of rosuvastatin in inhibiting atheromatous plaque and apoptosis in the plaque.Methods: Thirty healthy six-week old male mice were randomly divided into high fat diet group(n=10), rosuvastatin group(n=10) and general diet control group(n=10).The mice in first two groups were fed with high fat diet, and the mice in general diet control group were fed with general diet;4 weeks later the mice in three groups were respectively given 0.9% NaCl solution, rosuvastatin(10 mg·kg·d^-1) and 0.9% NaCl solution by gavage for 8 weeks.And then ELISA was used to detect the serum lipid levels, the area of atherosclerotic plaque was measured by HE staining, and the apoptosis in plaques was detected by TUNEL method;the expression of apoptosis-assoicated gene Bax protein was measured by immunohistochemical staining.Results: Compared with high fat diet group, the levels of total cholesterol(TC) and low density lipoprotein cholesterol(LDL-C) of the mice in rosuvastatin group were significantly decreased(P<0.05 or P<0.01), and the 1evel of high density lipoprotein cholesterol(HDL-C) was significantly increased(P<0.01).The aortic atherosclerotic plaque of the mice in high fat diet group was massive with a great quantity of foam cells, cholesterol crystal, necrotic cells and inflammatory cells in the plaque;the aortic atherosclerotic plaque of the mice in rosuvastatin group was smaller with less foam cells, necrotic cells and inflammatory cells; the aortic atherosclerotic plaque area of the mice in rosuvastatin group was significantly smaller than that in high fat diet group(P<0.01);the apoptotic index in the aortic atherosclerotic plaque of the mice in rosuvastatin group was significantly lower than that in high fat diet group(P<0.01).Compared with high fat diet group, the expression of Bax protein in rosuvastatin group was significantly decreased.Conclusion: Rosuvastatin may inhibit the progression of atherosclerotic plaque and the apoptosis in plaques through mitochondrial pathway.

Objective: To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs) for regeneration of tissue injuries and defects.Methods: Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently.Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2, B27, basic fibroblast growth factor(bFGF), and epidermal growth factor(EGF).The skin dermis derived spheroids(SDDSs) were collected and monolayer cultured in serum-containing medium.Finally, the cells were characterized by immunofluorescence staining and differentiation assays.Results: The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium.After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number.The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin.Under osteogenic, chondrogenic and adipogenic differentiation conditions, these cells were differentiated into the alizarin red, safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs.However, safranin O staining was weaker in the dermis derived cells than BMSCs.Conclusion: A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials, demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.

Objective: To investigate the effects of lipopolysaccharide (LPS) on the acute lung injury (ALI) and expressions of aquaporin 1 (AQP1) and aquaporin 5 (AQP5) in lung tissue of the rats. Methods: Forty-eight SPF grade male Wistar rats were randomly divided into control group and LPS group (n=24).The rats in LPS group were intravenously injected with LPS to induce ALI models, and the rats in control group were injected with saline.The rats were sacrificed at 2, 6, 12 and 24 h, and the samples were collected after the successful modeling.The pathological changes of lung tissue were observed with HE staining; the lung wet/dry weight (W/D) ratio and lung permeability index were detected; ELISA was used to detect the levels of TNF-α and MIP-1α.The expression levels of AQP1 and AQP5 protein and mRNA were measured by Western blotting, immunohistochemistry and Real-Time PCR methods.Results: Compared with control group, the TNF-α and MIP-1α levels in LPS group were significantly elevated at 2, 6 and 12 h (P<0.05), and at 24 h they were gradually reduced to the normal level.The HE staining results showed the alveolar and interstitial edema at 2 h after LPS injection, obviously in 12 h.The lung W/D ratios and pulmonary permeability indexes at different time points in LPS group were significantly higher than those in control group (P<0.05), and they reached the peak at 12 h.The expression levels of AQP1 and AQP5 mRNA and protein in lung tissue of the rats at different time points in LPS group were significantly lower than those in control group (P<0.01).Conclusion: LPS can induce ALI in the rats and down-regulate the expressions of AQP1 and AQP5; LPS is involved in the formation of pulmonary edema.

Objective: To observe the effects of ischemic postconditioning (I-postC) on the lung injury after limb ischemia reperfusion(LIR) in the rats, and to investigate the protective effect and the possible mechanisms.Methods: 24 Wistar rats were randomly divided into control group, ischemia-reperfusion group(IR group) and I-postC group(n=8).Referring to routine method in our department, the model rats which underwent 4 h ischemia and 4 h reperfusion of hind limbs were made.In control group, the rubber band around the limb was loose and the blood flow was not blocked.In I-postC group, before reperfusion, ischemia 5 min and reperfusion 5 min were performed in the rats, repeated for 3 times and then perfusion 4 h was taken, The blood and lung tissue from every rat were taken accurately.The percentages of CD18 positive cells in peripheral blood, the levels of soluble intercellular adhesion molecule-1(sICAM-1)and P-selectin in plasma, the myeloperoxidase (MPO) activities in lung tissue, the levels of intercellular adhesion molecule-1 (ICAM-1) and P-selectin in lung tissue of the rats in various groups were detected.The partial pressure of oxygen (PaO₂) and partial pressure of carbon dioxide (PaCO₂) were measured.The morphological changes of lung tissue under light and electron microscopes were observed.Results: Compared with control group, the percentage of CD18 positive cells and the levels of sICAM-1 and P-selectin of the rats in IR groups were increased(P<0.01);PaO₂ and PaCO₂ were decreased significantly;the MPO activity in lung tissue was also significantly increased(P<0.01).The HE staining results showed lung interstitial vascular dilation, congestion, PMN infiltration, the increased gap blood vessel, alveolar septal thickening, alveolar exudation, bronchial epithelial cell shedding and necrosis of the rats in IR group.Compared with IR group, the values of biochemical indicators mentioned above were decreased obviously(P<0.01);PaO₂ and PaCO₂ were increased significantly(P<0.01); the activities of inflammatory factors in plasma and lung tissue were decreased (P<0.01); the pathological changes of lung damage were improved significantly.Conclusion: I-postC can reduce the lung injury after LIR in the rats, and the mechanism may be related to inhibiting the inflammatory reaction.

Objective: To investigate the silencing of P27RF-Rho gene with lenvirus targeting mediated technique, and to clarify its influence in the invasion of liver cancer cells.Methods: The P27RF-Rho RNAi lentivirus was constructed.The liver cancer BEL7402 cells were infected with lentivirus.The experiment was divided into P27RF Rho-siRNA group, scramble-siRNA group and BEL7402 group. The effect of silencing P27RF-Rho gene and the expression levels of hepatocellular carcinoma (HCC) associated proteins RhoA, RhoC, VEGF, P53 and PTEN were detected;the activities of matrix metalloproteinase (MMPs)associated with tumor invasion were analyzed by Gelatin zymography; the variation of transfer ability and invasion abilities were compared by Wound healing assay experiment and Transwell experiment.Results: The Western blotting results showed the expression levels of P27RF-Rho, RhoA, RhoC, and VEGF proteins in the BEL7402 cells in experiment group were significantly lower than those in two control groups(P<0.05), and the expression levels of P53 and PTEN were higher than those in two control groups(P<0.05).The results of Gelatin zymography showed the activities of MMPs in experiment group were significantly lower than those in two control groups(P<0.01);Wound healing assay showed that the migration ability of the BEL7402 cells in experiment group was significantly inhibited (P<0.01);the number of cells passed through the Transwell Chambers in experiment group was significantly less than those in two control groups (P<0.01).Conclusion: Silenceing P27RF-Rho can weaken the invasion ability and migration ability of human HCC BEL7402 cells.

Objective: To explore the inhibitory effect of dihydroartemisinin (DHA) on the growth of neuroblastoma cells, and to clarify the anti-tumor mechanism of DHA.Methods: The experiment was divided into blank control group and DHA groups (the final concentrations of DHA were 0.05, 0.50, 5.00 and 50.00 μmol·L^-1).The proliferation rates of neuroblastoma SH-SY5Y cells after treated with DHA were examined by MTT assay; the changes of cell cycle of SH-SY5Y cells after treated with DHA were examined by flow cytometry;the expression levels of cyclin D₁ and caspase-3 proteins were detected by ELISA and Western blotting methods.Results: The proliferation of SH-SY5Y cells 24, 48, and 72 h after treated with different concentrations of DHA were inhibited.Compared with blank control group, the proliferation rates of SH-SY5Y cells in 0.50, 5.00 and 50.00 μmol·L^-1DHA groups were significantly decreased(P<0.05 or P<0.01).The density of cells was decreased with the increasing of DHA concentration.Compared with blank control group, the percentage of SH-SY5Y cells at SubG₁ phase in 50.00 μmol·L^-1 DHA group was increased(P<0.05), and the percentage of cells at G₀/G₁ phase was increased first then was decreased; otherwise, the percentages of cells at S and G₂/M phase were decreased.Compared with blank control group, the expression level of cyclin D1 protein in 50.00 μmol·L^-1 DHA group was decreased(P<0.05), but the expression level of caspase-3 protein in 50.00 μmol·L^-1 DHA group was increased(P<0.05).Conclusion: DHA could inhibit the proliferation through arresting the cell cycle and inducing the apoptosis of neuroblastoma cells.

Objective: To explore the influence of different cell culture methods in the genome-wide DNA methylation status of breast cancer MDA-MB-231 cells, and to clarify the relationship between genome-wide DNA methylation status and cell growth environment and the role of genome-wide DNA methylation status in the occurrence and development of tumor.Methods: The MDA-MB-231 cells were cultured with 2D and 3D cell culture models and mouse orthotopic transplantation model (Timodel) and collected, then DNA was extracted by DNA extraction kit and the genome-wide DNA methylation status of MDA-MB-231 cells after cultured with three different culture methods was detected by DNA methylation chip, then the value of beta, DiffScore and Delta_Beta of the CpG loci of each gene were calculated by applying Genomestudio software, and the differential methylation genes were screened by Genomestudio software and GO and Pathway analysis of these genes were performed in DAVID on-line analysis tool.Results: All 480 genes of the MDA-MB-231 cells showed significant differences in the degree of methylation in 3D and 2D models(P<0.05);86 448 genes in 3D and Ti models(P<0.05); 90 005 genes in Ti and 2D models(P<0.05).The differential methylation genes in 3D and 2D, 3D and Ti, and Ti and 2D models were enriched on the multicellular organismal development term and cell differentiation term(P<0.05);also on MAPK signaling pathway, cell adhesion molecules (CAMs), and regulation of actin cytoskeleton(P<0.05).Conclusion: There are differences in genome-wide DNA methylation status of MDA-MB-231 cells cultured in 2D, 3D cell culture and Ti models.

Objective: To explore the expression of pressure-sensitive transient receptor potential channel (TRPC1) in bronchial epithelium cells of the patients with chronic obstructive pulmonary disease(COPD), and to explain the molecular mechanism about the participation of TRPC1 in airway remodeling.Methods: Sixty-four patients who needed fiberoptic bronchoscope for diagnosis were selected.All the patients were used as non-operation group and divided into COPD group(40 cases of mild to moderate grade) and control group(24 cases without chronic airway inflammatory disease) according to Respiratory Disease Guideline.The level of the forced expiatory volume/forced vital capacity (FEV₁/FVC) in the first second and forced expiatory volume in the first second of the expected value (FEV₁% pred) were detected by lung function test.The expression levels of TRPC1, matrix metalloproteinase(MMP-9) and transforming growth factor-β₁(TGF-β₁) in bronchoalveolar liquid(BALF) were measured by ELISA.The relevance among expression level of TRPC1 and levels of FEV₁/FVC, FEV₁% pred, MMP-9, TGF-β1 was analyzed.Seventeen lung samples from the patients underwent pulmonary lobectomy were selected. All the patients were used as operation group and divided into COPD group(8 cases of mild to moderate grade) and control group(9 cases without chronic airway inflammatory disease). Then indictator relevance to airway remodeling, such as thickness-diameter ratio(TDR%), percentage of wall thickness(WA%) were measured.The immunohistochemistry experiment was used to detect the distribution characteristics of TRPC1 protein in human bronchial epithelial cells.The Imagepro-plus 6.0 software was used to analyze the expression difference of TRPC1 protein between COPD and control groups.Western blotting method was used to detect the expression level of TRPC1.The correlation between the expression level of TRPC1 and TDR%, WA% was analyzed.Results: The lung function detection results showed that in non-operation group, the levels of FEV₁% pred and FEV₁/FVC in COPD group were decreased compared with control group(P<0.05).The expression levels of TRPC1, MMP-9 and TGF-β1 in COPD group were increased compared with control group(P<0.05). The expression level of TRPC1 was negatively correlated with FEV₁%pred(r=-0.34, P=0.002) and FEV1/FVC(r=-0.38, P=0.004).The expression level of TRPC1 was positively correlated with MMP-9(r=0.36, P=0.004) and TGF-β₁ (r=0.61, P=0.002).The immunohistochemistry results in operation group displayed that the TRPC1 protein was mainly distributed in the epithelial columnar cell nucleus and the cell membrane near lumen within the bronchial epithelium.The ratio of integrated optical density(IOD) to area within the bronchial epithelium in COPD group was significantly higher than that in control group(P<0.05).The expression level of TRPC1 protein detected by Western blotting method in COPD group was higher than that in control group(P<0.05).The expression level of TRPC1 was positively correlated with TDR%(r=0.59, P=0.002) and WA%(r=0.60, P=0.002).Conclusion: The TRPC1 channel in human bronchial epithelial cells of COPD patients is actived and up-regulated.The expression level of TRPC1 channel is positively correlated with impared extent of lung function and the expression levels of cytokines relevant to airway remodeling.All above imply that pressure-sensitive TRPC1 channel could promote the development of airway remodeling.

Objective: To investigate the expression of CD56 antigen in leukemia cells of the patients with acute myeloid leukemia (AML) and its relationship with the prognosis of AML, and to clarify the role of CD56 antigen expression in predicting the prognosis of the AML patients.Methods: 171 AML (non-M3) patients aged from 14 to 60 years old, who received a IA Regimen as the first time inducing chemotherapy were chosen.Flow cytometric analysis was used to evaluate the CD56 expression in leukemia cells.COX proportional regression analysis was used to select the prognostic factors, and bivariable analysis was used to study the relationship between the positive rate of CD56 and overall survival (OS).The CD56⁺ group(n=52), including CD56≥50% expression group(n=39) and CD56<50% expression group(n=13), and CD56^- group(n=119) were identified by the expression of CD56 antigen.The complete remission rate (CRR), the relapse rate, the median OS, the median disease-free survival (DFS) and the survival rate of patients were compared.Results: The medium OS of the patients in CD56⁺ group (14.2 months) was shorter than that in CD56^- group (39.4 months) (P<0.05).Moreover, the medium OS in CD56≥50% group was shorter than that in CD56<50% group (11.7 months vs 20.3 months, P<0.05).The 1-year and 2-year survival rates of the patients in CD56⁺ group(61.5%, 46.2%) were lower than those in CD56^- group(75.6%, 63.9%)(P<0.05).The 1-year survival rate had no significant difference between CD56≥50% group and CD56<50% group (53.8% vs 84.6%, P>0.05), while the 2-year survival rate in CD56≥50% group was lower than that in CD56<50% group (41.0% vs 61.5%, P<0.05).There were no significant differences of the CRR between CD56⁺ group(76.9%) and CD56^- group (68.9%) as well as CD56≥50% group(58.9%) and CD56<50% group(63.5%) (P>0.05). The relapse rate and first year relapse rate of patients in CD56⁺ group (64.3% and 37.5%) were significantly higher than those in CD56^- group (34.3% and 17.9%) (P<0.05).However, there were no significant differences of the relapse rate and first year relapse rate between CD56≥50% group(75.0% and 42.9%) and CD56<50% group(37.5% and 16.7%) (P>0.05).The DFS in CD56⁺ group was shorter than that in CD56^- group (P<0.05).The same DFS result was also found between CD56≥50% group and CD56<50% group(P<0.05).Conclusion: The expression of CD56 antigen in leukemia cells predicts a bad prognosis in the AML patients, and the higher expression of CD56 indicates the worse prognosis.

Objective: To detect the expression levels of neuropilin1(NRP1) mRNA and miR-9 in non-small cell lung cancer (NSCLC) tissue samples, and to explore the correlations between the expressions of NRP1 mRNA, miR-9 and the clinicopathological characteristics of the patients with NSCLC. Methods: Informed consent was obtained from each patient before surgery.The tissue samples including 45 NSCLC tissue, 45 adjacent carcinoma tissue and 45 normal lung tissue were collected from China-Japan Union Hospital of Jilin University from 2010 to 2011. qRT-PCR was used to detect the expression levels of NRP1 mRNA and miR-9 in three kinds of lung tissue, and the correlation between the expressions of NRP1 mRNA, miR-9 and clinicopathological characteristics of the patients with NSCLC was analyzed.Results: Compared with normal tissue, the expression level of NRP1 mRNA in adjacent carcinoma tissue had no change(P>0.05), but the expression level of NRP1 mRNA in non-small cell lung cancer tissue was significantly decreased (P<0.05).Compared with normal tissue, the expression level of miR-9 in adjacent carcinoma tissue had no change(P>0.05), but the expression level of miR-9 in non-small cell lung cancer tissue was significantly increased (P<0.05).Furthermore, in adjacent carcinoma tissue, the expression level of miR-9 in the males was lower than that in the females(P<0.05).In NSCLC tissue, the expression level of NRP1 mRNA had no relationship with sex, age, differentiation degree, TNM stage and clinical stage, but was significantly correlated to the histological subtype and lymph node metastasis (P<0.05). In NSCLC tissue, the expression level of miR-9 had no relationship with age, pathological type, lymph node metastasis, differentiation degree, TNM stage, and clinical stage (P>0.05), but was correlated to the sex(P<0.05).Conclusion: The expression level of miR-9 is up-regulated and the expression level of NRP1 mRNA is down-regulated significantly in non-small cell lung cancer tissue.The detection of the expression level of NRP1 mRNA contributes to judge the histological subtype and lymph node metastasis of NSCLC.

Objective: To evaluate the diagnostic values of single and combined detection of serum CA199, complement 3 (C3), complement 4 (C4), total cholesterol (TC), triglyceride (TG), and lipid metabolism levels in the patients with pancreatic cancer, and to explore their correlations with TNM stage and pathological stage of pancreatic cancer.Methods: Total 185 subjects were enrolled into the study by three groups:pancreatic cancer patients group(Pc group, n=77), non-digestive system cancer patients group (Ndc group, n=58) and healthy control group (Hc group, n=50).The levels of serum CA199, C3, C4, and high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol(LDL-C), apolipoprotein A (ApoA), apolipoprotein B (ApoB), apolipoprotein E (ApoE), and lipoprotein a (Lpa) levels were detected.Results: ① The serum level of CA199, C3, C4, and ApoE of the patients in Pc group were higher than those in Ndc and Hc groups (P<0.01).No statistical difference was observed in the serum CA199 between Ndc and Hc groups(P>0.05). The levels of C3, C4, and ApoE in Pc group and Ndc group was higher than those in Hc group (P<0.01), and the levels of the biomarkers in Pc group were also higher than those in Ndc group (P<0.01).The levels of HDL-C, ApoA and Lp(a) of the patients in Pc group were significantly lower than those in Ndc and Hc groups (P<0.05).② The area under ROC curve (AUC) of serum CA199, C3, C4, ApoE, HDL-C, and ApoA were 0.916, 0.841, 0.788, 0.785, 0.834, and 0.810, respectively.Furthermore, multiple factor analysis showed that the combined detection of CA199, C3, and HDL-C (AUC=0.968) improved the diagnosis compared with detecting CA199 alone (P<0.05).① The CA199 level of the patients in Ⅲ-Ⅳ stage of TNM stage was higher than that in the patients in Ⅰ-Ⅱ stage (P<0.01).For the pathological stage, the ApoA level in low differentiation group was higher than that in moderate and high differentiation group (P<0.05).There was no statistical difference in other biomarkers between the different TNM stages and pathological stages.Conclusion: The levels of CA199, C3, C4 and ApoE of pancreatic cancer patients are significantly increased, while the levels of HDL-C, ApoA, and Lp (a) are significantly reduced.Combined detection of CA199, C3, and HDL-C can improve the early diagnosis of pancreatic cancer compared with the single assessment of each biomarker.

Objective: To determine the derivative chromosome 9 by the method of detecting the ASS gene, and to explore the relationship between the deletion of derivative chromosome 9 and the efficacy and prognosis of the chronic myeloid leukemia (CML) patients.Methods: The materials of 34 CML patients with positive BCR-ABL fusion gene whose ASS genes were detected were retrospectively analyzed. All patients were treated with Extra-signal (ES) probe to detect the derivative chromosome 9.All patients were divided into two groups according to whether they carried the derivative chromosome 9.The blastic phase or theaccelerated phase rates in two groups were compared by using Fisher exact probability.Results: All patients were detected by FISH (BCR-ABL ES probe), and all the BCR-ABL fusion signals were positive.6 of 34 patients were found the deletion of ASS gene, among them 1 case blonged to chronic phase, and 5 cases developed into blastic phase or accelerated phase.In the patients without ASS gene deletion, there were 22 cases in chronic phase, and 6 cases in plastic phase or accelerate phase, there was significant difference of blastic phase rate/accelated phase rate between them(P<0.05).A total of 26 patients were treated with tyrosine kinase inhibitors (TKI). 5 of 26 patients belonged to the ASS gene deletion group, 1 of 5 patients treated with TKI got molecular remission, 4 of 5 patients developed into blastic phase or accelerated phase. 21 of 26 patients belonged to the group without ASS gene deletion, and among them, 19 cases got molecular remission, 2 cases developed into plastic phase or accelerated phase after treatment of TKI, there was significant difference between them(P<0.05).6 patients were treated with traditional chemotherapy (hydroxyurea, interferon);1 of 6 patients belonged to the ASS gene deletion group, finally developed into the blastic phase or accelerated phase; 5 of 6 patients belonged to the group without ASS gene deletion, 2 cases got the hematological remission, and 3 patients developed into blastic phase or accelerated phase after treatment, and there was no significant difference of blastic phase rate/accelerated phase rate between them (P>0.05).Conclusion: The CML patients with derivative chromosome 9 (ASS gene deletion) prone to get disease progression, and have a higher proportion in the blastic phase or accelerated phasepatients.Derivative chromosome 9 is related to the bad treatment efficacy of TKI and the poor prognosis of CML.

Objective: To combine the detection of serum levels of adenosine deaminase(ADA)and T-cell spot test(T-spot.TB), and to explore their significances in diagnosis of pulmonary tuberculosis.Methods: 159 patients suspected with pulmonary tuberculosis were selected and divided into pulmonary tuberculosis group and non-tuberculosis group(n=68); 80 healthy people were randonly selected as healthy control group. The serum ADA levels and number of T-spot of the subjects in three groups were detected. Ther serum ADA levels and the positive rates of T-spot.TB in various groups and their sensitivities and specifities were compared.Results: The serum ADA level of the patients in pulmonary tuberculosis gruop was (22.10±6.60)U·L^-1;those in non-tuberculosis group and healthy control group were (16.90±6.35) and (8.70±5.98)U·L^-1;the serum ADA level in pulmonary tuberculosis group was significantly higher than those in non-tuberculosis group and heathy control group(P<0.05).The positive rate of serum ADA level in diagnosis of pulmonary tuberculosis was 56% and the T-spot.TB positive rate in diagnosis of pulmonary tuberculosis was 87.9%.Combined use of parallel test, the detection sensitivity was 91.2%; using the series of joint tests, the specificity was 94.6%.Conclusion: Combined detection of serum level of ADA and T-spot.TB can significantly improve the clinical diagnosis efficacy of pulmonary tuberculosis.

Objective: To investigate the correlation between the CTLA-4 expression in the peripheral blood CD4⁺ and CD8⁺ T cells in the patients with glioma and glioma WHO classification and operation, and to clarify the clinical significancesof CTLA-4.Methods: 60 patients with glioma from our hospital were selected as glioma group, and 46 healthy volunteers were used as control group.Then the CTLA-4 expression levels in the peripheral blood CD4⁺ and CD8⁺ T cells of the subjects in glioma group and control group were detected by fluorescence-activated cell sorting (FACS) and the relationship between CTLA-4 and glioma WHO classification and surgery was analyzed.Results: The CTLA-4 expression levels in CD4⁺ and CD8⁺ T cells in peripheral blood of the patients in glioma group were higher than those in control group(P<0.01).The CTLA-4 expression level in the patients with grade Ⅳ glioma was the highest, the expression level of the patients with grade Ⅳ glioma was higher than that of the patients with grade Ⅲ glioma(P<0.01), and the expression level of grade Ⅲ the patients with was higher than that of the patients with grade Ⅱ(P<0.01).The expression level of CTLA 4 in CD4⁺ and CD8⁺ T cells of the patients with glioma after operation was lower than beforeoperation(P<0.01).Conclusion: The CTLA-4 expression levels in the peripheral blood CD4⁺ and CD8⁺ T cells are increased with the increasing of malignancy degree of the patients with glioma, and it has a promotion role in the occurrence of development of glioma.

Objective: To investigate the influence of single nucleotide polymorphisms(SNP) of cytidine deaminase(CDA) in the curative effect of gemcitabine, and to clarify the correlation of CDA SNP with the clinical prognosis of the advanced non-small cell lung cancer(NSCLC) patients.Methods: CDA 79A>C gene was detected by Sequenom Massarray Genotype System among 93 patients with advanced NSCLC who received chemotherapy of gemcitabine.The associations between CDA 79A>C polymorphisms and progression free survival(PFS) and overall survival(OS) were estimated using Kaplan-meier methods and Log-rank test for univariate analysis, and COX proportional hazards model was used for multivariate analysis.Results: There was no significant correlation between the CDA 79A>C SNPs with the clinical and pathological characteristics of NSCLC patients.The median PFS in the patients with wild genotype A/A was 9.3 months, while the median PFS in the patients with mutant genotype A/C and C/C was 11 months;there was no significant difference between them (P=0.061). The median OS in the patients with wild genotype A/A was 21.7 months, and the median OS of the patients with mutant genotype was 22.1 months;there was no significant difference between them (P=0.513).Conclusion: CDA 79A>C SNPs can prolong the PFS of the NSCLC patients, but it does not affect the OS of the patients.

Objective: To determine the inflammatory factor levels and inflammatory cell percentages in the patients with cough variant asthma (CVA), and to clarify their potential role in the pathogenesis of CVA unresponsive to bronchodilator treatment.Methods: 60 patients with CVA were randomly selected and divided into CVA unresponsive to bronchodilator treatment group (n=30) and CVA responsive to bronchodilator treatment group (n=30).As the same time 30 cases of healthy persons were used as normal control group.The levels of interluekin-8(IL-8) and esoinophil cationic protein (ECP) in their induced sputum were detected, the classification of inflammatory cells in their induced sputum were observed, and their scores of cough symptom were recorded.Results: The IL-8 level in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group was higher than that in CVA responsive to bronchodilator treatment group and normal control group (P<0.05).The ECP level in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group was lower than that in CVA responsive to bronchodilator treatment group (P<0.05), but it was similar to the level in normal control group (P>0.05).The neutrophil percentages in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group were higher than those in CVA responsive to bronchodilator treatment group and normal control group (P<0.05).The scores of cough symptom of the patients in CVA unresponsive to bronchodilator treatment group was positively correlated with IL-8 level(r=0.764, P<0.01), and the scores of cough symptom of the patients in CVA unresponsive to bronchodilator treatment group was positively correlated with the neutrophil percentage in induced sputum(r=0.889, P<0.01).Conclusion: IL-8 and neutrophil may be associated with the incidence of CVA unresponsive to bronchodilator treatment.They can aggravate the inflammation and hypersensitivity of airway and cough symptom.The determination of IL-8 and neutrophil can be used as an accessory method in the diagnosis and judgement of severity degree and curative effect of CVA in clinic.

Objective: To compare the differences of lung function parameters of the patients with congenital scoliosis(CS) or idiopathic scoliosis (IS), and to clarify their influence in respiratory function.Methods: Sixty-one patients of scoliosis undergoing operation were selected.According to the cause they were divided into 25 patients with CS and 36 patients with IS.The changes of lung function parameters and the differences in the patients with scoliosis were analyzed. Results: The ages, body weights, Cobb angles of the patients in two groups had no significant differences.There were 6 cases and 12 cases of ventilation disturbance in CS and IS groups, and 15 cases and 25 cases of ventilation dysfunction.The levels of residual volume(RV), ratio of residual volume to total lung capacity(RV/TLC), RV/TLC percent predicted(RV/TLC% pred) in two groups were higher than the normal. The percentage of forced vital capacity predicted(FVC%pred) and FEV₁/FVC in CS group were both lower than those in IS group (P<0.05).Some pulmonary function parameters such as vital capacity(VC%pred), FVC%pred, forced expiratory volume in one second(FEV₁%pred), maximal mid expiratory flow(MMEF%pred), maximal voluntary ventilation(MVV%pred), TLC%pred and diffusion capacity for carbon monoxide of the lung(DLCO%pred) were negtively correlated with Cobb angle(r=-0.54--0.35, P<0.05) in IS group and the TLC%pred was negtively correlated with Cobb angle(r=-0.047, P<0.05) in CS group.Conclusion: Different types of scoliosis have influence in the pulmonary function of the patients.The degree of pulmonary function injury of the patients with IS is worse than that of the CS patients.The IS patients need treatment in time.

Objective: To explore the efficacy and safety of aprepitant combined with tropisetron and dexamethasone in the prevention of emetogenic chemotherapy-induced nausea and vomiting and clarify its influence in the life guality of patients, and to evaluate its safety in treatment.Methods: One hundred and fifty six patients with malignant tumor were randomly assigned to observation group (n=75)(day 1, aprepitant 125 mg, tropisetron 4 mg and dexamethasone 6 mg 1 h before chemotherapy, day 2-4, aprepitant 80 mg and dexamethasone 3.75 mg, qd) and control group (n=81) (day 1, tropisetron 4 mg and dexamethasone 6 mg before chemotherapy, day 2-4, dexamethasone 3.75 mg, qd). The primary study endpoints were the comparisons of acute and delayed complete response rates of nausea and vomiting, and the second study endpoints were the evaluations on the improvement of the life quality of patients.Results: General evaluation on the primary study endpoints was performed, The complete remission rate in observation group was 69.3%(52/75), and the complete remission rate in control group was 53.1%(43/81), there was signficant difference between two groups(P=0.049). For the acute nausea and vomiting(0-24 h), the complete remission rate in observation group was 78.7%(59/75), and the complete remission rate in control group was 75.3%(61/81), and there was no significant difference between two groups(P=0.705). The complete remission rate of delayed nausea and vomiting(25-120 h) in observation group was significantly higher than that in control group (76.0% vs 54.3%)(P=0.007). For the second study endpoints, the average scores of FLIE in observation and control group were 120.8±12.4 and 84.0±8.7, and there was significant difference between two groups(P<0.05).Conclusion: Aprepitant can significantly prevent the emetogenic chemotherapy-induced nausea and vomitting and improve the life quality of patients.

Objective: To systematically review the effects of statins in the treatment of the patients with pneumonia or sepsis, and to provide evidence-based basis for the using of statin in treatment of pneumonia or sepsis.Methods: PubMed, Embase, The Cochrane Library, CNKI, WanFang Data and VIP were searched for randomized placebo-controlled trials of statins published from the establishment time of datebases to January 31, 2015.The qualities of the included studies were independently assessed by two reviewers and the relevant data was extracted for analysis using Review Manager 5.3.Results: Nine trails involving a total of 1227 patients were included.①A total of 6 studies about the mortality of hospitalized patients were included in the Meta-analysis;compared with placebo group, statins didn't improve the mortality of hospitalized patients with pneumonia or sepsis; risk ratio (RR)=0.83, 95%CI(0.63, 1.08), P=0.17.②A total of 7 studies about the rates of mechanical ventilation usage or ICU admission of the patients were included in the Meta-analysis.Compared with placebo group, statins didn't improve the rates of mechanical ventilation usage or ICU admission of the patients with bacteria infection; RR=0.99, 95% CI(0.96, 1.03), P=0.62.③A total of 4 studies about the levels of C reaction protein (CRP) were included in the Meta-analysis.Compared with placebo group, statins significantly reduced the levels of CRP; mean differences(MD)=-8.30mg·L^-1, 95%CI(-12.13, -4.47), P<0.0001.Conclusion: Statins can not significantly reduce both the in-hospital mortality and the rates of mechanical ventilation usage or ICU admission, but can significantly reduce the plasma CRP levels, and reduce the inflammation.

Objective: To explore the regular variation pattern of tumor volumes of the patients with non-small cell lung cancer(NSCLC)before and after targeting treatment of epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI), and to clarify its clinical value.Methods: The materials of 39 NSCLC patients with EGFR-TKI targeting treatment were retrospectively analyzed. The tumor volumes were detected by volume measurement software of TPS and Image J image processing software, then the absolute and relative tumor volume changes of the NSCLC patients before and after targeting treatment were analyzed by paired sample comparison symbol Wilcoxon rank test. Results: The absolute tumor volumes(mm³) of the patients with NSCLC before and 1 month after targeting treatment were 14822.11(7524.73, 54999.41)and 7954.42(3499.73, 29396.83), respectively, and there was statistically significant difference(Z=-3.257, P=0.001);the absolute tumor volumes of the patients with NSCLC 1 and 2 months after targeting treatment were 8 358.47(4394.36, 24 430.05)and 7028.76(3634.98, 21056.71), respectively, and there also was statisticaliy significant difference(Z=-2.213, P=0.027).When the original tumor volume before targeting treatment was regarded as 1, the relative tumor volume of 1 month after targeting theatment was 0.612 6(0.313 8, 0.853 7), and there was significant difference(Z=-3.855, P<0.001);the relative tumor volumes of 1 month and 2 months after targeting treatment were 0.6084(0.3643, 1.0443) and 0.423 0(0.248 8, 0.877 7), respectively, and there also was statistically significant differernce(Z=-2.173, P=0.030);but the differences between other consecutive months(from 3 months to 6 months) had no statistically significant differences(P>0.05);the changes of tumor relative volume presented platform stage after 3 months.The tumor relative volumes of 7-9 months after EGFR-TKI treatment reached the bottom. Conclusion: The average primary tumor volume of the NSCLC patients is obviously reduced 1 and 2 months after TKI targeting treatment.It may be optimal to carry out radiotherapy in 3-9 months after EGFR-TKI targeting treatment.

Objective: To assess the efficacy of trastuzumab combined with neoadjuvant chemotherapy in the treatment of human epidermal growth factor receptor-2 (Her-2) positive breast cancer patients, and to explore its influencing factors of prognosisof breast cancer.Methods: The clinical materials of 112 Her-2 positive breast cancer patients were collected and they were divided into combined treatment group (trastuzumab+neoadjuvant chemotherapy) and single chemotherapy group (without trastuzumab).SPSS 19.0 software was employed to calculate and analyze their clinical characteristics.The survival rate and prognosis were analyzed by Kaplan-Meier method, Log-rank test, and Cox regression.Results: Among the 112 Her-2 positive breast cancer patients, 23 cases were treated with trastuzumab, 89 cases were treated without trastuzumab.The disease-free survival (DFS) in combined treatment group and single chemotherapy group had significant difference (P=0.012).And there was no significant difference of overall survival (OS) between two groups (P=0.064).The HR negative group had 18(32.7%) patients with 5-level of Miller and Payne (MP) classification which was higher than HR positive group(5/46, 10.9%) (P=0.009).In univariate analysis, the tumor size, node status at diagnosis and node status after operation were the influencing factors of DFS in the Her-2 positive breast cancer patients; the tumor size, node status at diagnosis, MP classification and node status after operation were the influencing factors of OS in the Her-2 positive breast cancer patients.The result of multivariate analysis indicated that the tumor size, node status at diagnosis, and with or without radiotherapy were the independent influencing factors of DFS and OS in the Her-2 positive breast cancer patients.Conclusion: The breast lesions in the Her-2 positive breast cancer patients with negative HR are relieved after treated with trastuzumab combined with neoadjuvant chemotherapy.The combination of trastuzumab and neoadjuvant chemotherapy can significantly improve the DFS and has a positive influence in the prognosis of Her-2 positive breast cancer patients.

Objective: To investigate the clinlic therapeutic effect of sigmoid orthotopic neobladder after radical cystectomy in the treatment of invasive bladder cancer.Methods: 16 patients with invasive bladder cancer(all males;age range from 53 to 75 years)were admitted and underwent radical cystectomy and sigmoid orthotopic neobladder.Results: Sixteen patients were hospitalized for 37 to 62 d, with an average of 51 d.All patients were followed up for a mean of 26 months(range from 5 months to 6 years).The serum creatinine(Cr) and blood urea nitrogen(BUN) levels were both in the normal range and there was no acidosis in all the cases.No case had unilateral ureteral urine reflux during cystography.Of the 16 patients 14 cases(87.5%)achieved continence during daytime;6 cases(37.5%)were incontinent at night, but all of them could control urination by being woken up at night.The capacity of the neobladder was 245 to 380 mL with an average of 316 mL and the maximal pressure of the neobladder during filling was 28 to 57 cm H₂O (1 cm H₂O=0.098 kPa) with an average of 39 cm H₂O.Conclusion: As an operation the sigmoid orthotopic neobladder can be performed easily without serious postoperative complications and has more reliable results.This operation may be generally applied in clinical practice.

Objective: To explore the risk factors of delayed traumatic intracranial hematoma (DTICH) followed by unilateral large decompressive craniectomy (LDC) and its influence in the prognosis of the severe traumatic brain injury (sTBI) patients, and to improve the successful rate of the rescue.Methods: The clinical data of 130 sTBI patients underwent unilateral LDC were retrospectively analyzed. The patients were divided into DTICH group(n=42) and non-DTICH group(n=88) according to whether the DTICH occurred after operation. The risk factors of DTICH and its influence were contrastively analyzed.Results: The analysis results of the clinical data of patients in two groups showed that preoperative GCS, time from trauma to operation, skull fracture, midline shift > 1 cm, basal cistern disappear, activated partial prothrombin time (APTT), fibrinogen (FIB), and thrombin time (TT) were significantly correlated with the appearance of DTICH (P<0.05). Multivariable Logistic regression analysis showed that the time from trauma to operation, skull fracture, basal cistern disappearing and FIB were the independent risk factors of DTICH (P<0.05). The analysis results of GOS 3 months after operation of the patients in two groups showed that the prognosis of the patients in DTICH group was significantly worse than that in non-DTICH group (P<0.01).Conclusion: For those patients who accompanied with shorter time from trauma to operation, skull fracture, basal cistern disappearing and FIB decrease, the appearance of DTICH should be paid attention. DTICH can affect the prognosis of patients; prevention and early diagnosis are crucial to improve the prognosis of patients.

Objective: To report one case of hilar hepatic tumor treated by ultrasound-guided percutaneous puncture ¹²⁵I radioactive seed implantation, and to review the associated literatures.Methods: One patient's CT and ultrasonic examinations were taken, and the plan of TPS was taken. The patient was treated by ultrasound-guided percutaneous puncture ¹²⁵I radioactive seed implantation according to the plan of TPS. The needles were parallel, and the distance between the seeds were 1.0 to 1.5 cm.Results: The operation was successfully completed. No postoperative complications were encountered during 1 month follow up. The volume of tumor was obviously reduced 1 month after operation detected by ultrasound and CT.The local control effect of tumor was good.Conclusion: Ultrasound-guided percutaneous puncture ¹²⁵I radioactive seed implantation is a safe and effective method in treating hilar hepatic tumor.

Objective: To evaluate the value of adenosine triphophate (ATP) stress/rest nuclide myocardial perfusion imaging(MPI) in the diagnosis of female patients with coronary heart disease(CHD).Methods: The clinical materials of 47 female suspected CHD patients were retrospectively analyzed, aged from 39 to 74 years, and the average age was (53.7±6.3) years old.All patients were hospitalized and underwent two-day ATP stress and rest nuclide MPI and coronary angiography(CAG) in two weeks.The results and images of MPI and CAG were evaluated by more than 2 attending physicians.Using CAG as the "gold standard", the diagnostic efficiency (sensitivity, specificity and accuracy) of MPI for CHD was evaluated.Results: Compared with CAG, the sensitivity, specificity and accuracy of ATP stress MPI in diagnosing the female CHD patients were 81.3%(13/16), 77.4%(24/31)and 78.7%(37/47) individually;the positive predictive value and negative predictive value were 65.0%(13/20)and 88.9%(24/27).There were no severe adverse effects in the ATP stress test and the incidence of adverse effects was 85.1%.Conclusion: There is a highly diagnostic efficiency of ATP stress MPI in the CHD patients.It can be the first choice of examination methods for screening without injury and diagnosing the myocardial ischemia in the female patients.

Objective: To investigate the optimal time of coronary computed tomography angiography(CCTA)after taking nitroglycerine, and to provide basis for improving the image quantity of CCTA.Methods: 43 patients underwent CCTA were scanned with coronary artery calcium score (CACS) after taking nitroglycerin 0, 3, 5 and 10 min.Then the diameters of the same coronary artery from the same anatomic location and the expanding rates were measured, and the change regular was analyzed with single factor analysis of variance of SPSS 17.0 software.Results: The average coronary artery expanding rate was 8% 3 min after taking nitroglycerin, and the difference was significant compared with 0 min (P<0.05); but there were 5 patients whose coronary artery didn't expand.The coronary artery expanded obviously 5 and 10 min after taking nitroglycerin compared with 0 and 3 min (P<0.05); the maximum expanding rates at 5 and 10 min were both 38%.There were 13 and 28 patients whose coronary artery reached the maximum expanding rates (17% and 19%) 5 and 10 min after taking nitroglycerin, respectively; the coronary artery of 2 patients expanded at a same level at 5 and 10 min.The average expanding rates of coronary artery had no significant differences between 5 and 10 min (P>0.05).Conclusion: Taking nitroglycerin can significantly expand the diameter of coronary arteries.It is necessary to perform CCTA during 5-10 min after taking nitroglycerine.

Objective: To explore the clinical application values of contrast-enhanced ultrasound in the diagnosis and classification of portal vein tumor thrombosis(PVTT)by comparing with enhanced CT.Methods: 43 patients with PVTT confirmed by clinic and pathology were selected, and the accuracy rates of the diagnosis and clinic classification of contrast-enhanced ultrasound and enhanced CT were compared.Results: The characteristic of PVTT in contrast-enhanced ultrasound was "quick in fast out";88.4% (38/43) of PVTT were hyper-enhancement in the arterial phase, 81.4% (35/43) of PVTT were hypo-enhancement in the vein phase, and all tumor thrombosis showed hypo-enhancement in the delay phase.The diagnostic accuracy rate of contrast-enhanced ultrasound in PVTT was 100%, and the accuracy rate of enhanced CT was 97.7% (42/43).The classification accuracy rate of contrast-enhanced ultrasonic was 95.3% (41/43), and the accuracy rate of enhanced CT was 93.0% (40/43);there was no statistically significant difference between two methods(P>0.05).Conclusion: Dynamic contrast-enhanced ultrasonography can display the blood perfusion characteristics of PVTT, and displays the actual infiltrating tumor thrombosis.Ultrasonic imaging and enhanced CT in the qualitative diagnosis of PVTT and clinical classification have a good consistency.Contrast-enhanced ultrasound can be used as an important imaging method to evaluate the PVTT before treatment.

Objective: To explore the value of MR-cine for assessment of the duodenum peristalsis in the patients with functional dyspepsia. Methods: 25 patients with functional dyspepsia were selected according to the diagnostic criteria of Rome Ⅲ of functional gastrointestinal disorders(FGIDs) as case group and 25 cases of healthy volunteers who had been screened out in clinic were used as normal control group.After fasting for 8 h, supine position was performed.1.5TGEHDxMR was used to scan the coronal, axial and oblique coronal fast steady state precession (FIESTA) sequence of gastric and duodenal descending part.The images of 5 and 15 min after drinking 600mL mannitol solution (concentration 2.5%) were collected and sent to MR AW4.4 workstation.The duodenal motility in the patients with functional dyspepsia was measured by recording the times of duodenum and measuring and calculating the percentage occlusion of duodenum contractions (PDC).Results: After drinking mannitol, the oblique coronal scan of all subjects clearly displayed the anatomical structure and peristaltic wave of descending part of duodenum. After drinking 5 and 15min of mannitol, the peristaltic wave frequencies in case group were lower than those in normal control group. The descending duodenum PDC of the patients in case group was significantly lower than that in normal control group(P<0.01) 5 min after drinking. The PDC in normal control group 15 min after drinking was lower than 5 min (P<0.05); the duodenal PDC 15 min after drinking of mannitol had no statistical difference between case group and normal control group (P>0.05).Conclusion: MR-cine can evaluate preliminarly the duodenum peristalsis of the patients with functional dyspepsia.The diagnosis of FGIDs can be further studied by using the noninvasive MR-cine examination technique.

Objective: To analyze the levels of arachidonic acid(AA) and docosahexaenoic acid (DHA) in the breast milk of lactating mothers in Changchun City of Jilin Province, and to explore their influence factors.Methods: The lactating mother's basic information was collected with questionnaire, and the breast milk of lactating mothers on postpartum 22-25 d was obtained and the 3-day 24-hour dietary recall method was used to investigate the dietary intake information of 514 healthy lactating mothers.The Food Composition Table of China 2009 was used to calculate the intakes of five kinds of polyunsaturated fatty acids (PUFAs)in diet of lactating mothers and the gold key nutrition expert system software for corresponding nutrient analysis was used to calculate the amount of various kinds of foods in the lactating mothers' daily diet.The levels of AA and DHA in breast milk were determined with gas chromatography and the linear regression was used to analyze the related factors of AA and DHA levels in breast milk.Results: ①The average concentration of AA in breast milk of 514 lactating mothers was (0.08±0.04)g·100 g^-1, and the average concentration of DHA was(0.05±0.04)g·100 g^-1.②The single factor correlation analysis results showed that the oil intake was both positively correlated with AA and DHA levels in milk of lactating mothers(r=0.360, r=0.354, P<0.001), while the intakes of linoleic acid(LA), alpha linolenic acid(ALA), eicosapentaenoic acid(EPA), DHA, dairy and meats and seafood in diet were both negatively correlated with AA (r=-0.321, r=-0.280, r=-0.255, r=-0.299, r=-0.196, r=-0.306, P<0.05)and DHA(r=-0.315, r=-0.279, r=-0.175, r=-0.189, r=-0.248, r=-0.142, P<0.05).③The linear regression analysis results showed regression equation that dairy intake(β=-0.265) and EPA intake(β=-0.144) were both negatively correlated with the level of AA(P=0.009), and dairy intake was also negatively correlated with the level of DHA(β=-0.233, P<0.001).Conclusion: The AA and DHA levels in breast milk of lactating mothers didn't increase with the increasing of intake of milk or dairy products in the study.Moreover there is a competitive relationship between n-6 and n-3 series polyunsaturated fatty acids in the process of metabolism.

Objective: To investigate the influence of type A behavior in the occurrence and development of chromic periodontitis and compare the differences among people with different behavior types in the occurrence and development of chronic periodontitis, and to explore the influence of psychological factors in the process of periodontitis.Methods: According the same standard, 132 patients with periodontitis were selected as periodontitis group, and 126 patients without periodontitis were used as control group. Oral Surveys Basic Methods proposed by WHO in 1987 and Reference Standard of Diagnosis and Treatment of Periodontitis made by AAP in 2000 were used to dignose the periodontitis.All subjects finished type A behavior and SCL-90 questionnaire(ZHANG Boyuan 1983, revision in 1985), their scores were recorded and the results were analyzed by t test.Results: The proportion of type A behavior in periodontits group(68.94%) was higher than that in control group(27.78%) (P<0.05) compared with type M and type B behavior.The scores of hostility(1.54±0.38), anxiety(1.47±0.39), depression(1.41±0.37), interpersonal sensitivity (1.23±0.39), compulsive (1.72±0.46), and somatizition(1.47±0.38) were significantly higher than those in control group (1.32±0.30, 1.29±0.24, 1.25±0.23, 1.04±0.17, 1.41±0.35, 1.25±0.24). The calculus index(CI) of the people with type A behavior was higher than those of the people with other types of behavior(P<0.05).Conclusion: The people with tyep A behawior is easier to get periodontitis than other people.

Objective: To study the influence of different dietary patterns in hypertension, and to provide references for establishing a healthy dietary pattern, improving the abnormal blood pressure, preventing the early hypertension and controlling the high blood pressure.Methods: Random cluster sampling was adopted to analyze the dietary, examine the physical situations, and measure the blood pressures among 1 394 residents of Han nationality and Korean nationality aged from 40 to 80 years old from 8 villages in Jinlin Province from 2010 to 2011.Factor analysis was used to establish the dietary patterns and to analyze the effects of different dietary patterns on the prevalence of hypertension.Results: The prevalence of hypertension of middle aged and elderly people in the rural areas of Jilin Province was 35.3%.4 dietary patterns were obtained through factor analysis, including salted product dietary pattern, alcohol dietary pattern, condiments dietary pattern and high protein dietary pattern.By multivariate Logistic regression analysis and adjustment of the age, the gender, the nationality, the education and the career background, alcohol dietary pattern was positively correlated with the occurrence of the hypertension (OR=1.43, 95%CI 1.07-1.90). Conclusion: Alcohol dietary pattern is closely related to the hypertension.Long-time large amount of alcohol is a risk cause for high blood pressure.Reducing alcohol consumption and developing healthy diet habits can help to prevent hypertension.