Objective To analyze the polymorphism of thymidylate synthase(TS)in colorectal cancer patients,further more,to provide guidance for personalized therapy of colorectal cancer.Methods PCR direct sequencing was used to detect the polymor-phism of TS in 252 patients with colorectal cancer.Results TS genotypes of 252 patients with colorectal cancer were detected total-ly,including 137 male and 115 female.3RG/3RC accounted for the largest proportion in both male and female(36.50% and 36.52%respectively).In female,2RC/3RC and 3RG/3RG both accounted for the second largest proportion(both 18.25%).While in fe-male,3RG/3RC accounted for the second largest proportion(26.09%).If patients were divided according to age groups,in youth patients(n=28),3RG/3RC accounted for the largest proportion(42.86%),and the second was 2RC/3RG(21.43%).In the middle aged patients(n=84),3RG/3RC(45.24%)and 2RC/3RC(16.67%)were the major genotypes.For old patients(n=115),the ma-jor genotypes were 3RG/3RC(36.52%)and 2RC/3RG(16.52%).Conclusion The polymorphism of TS are mainly 3RG/3RC in colorectal cancer patients.

Objective:To prepare and purify recombinant human zona pellucida 3 protein,and to study its immunologic activity.Methods:The E.coli BL21 containing recombinant plasmid pGEX4T-1/ huZP3 was induced to express GST-fusion protein by IPTG.After a series of purification procedure,the purified protein was analyzed by SDS-PAGE.After the mice immunized with rhuZP3,the antibody responses against rhuZP3 were detected by ELISA.Results:The soluble fusion protein was expressed,and purity of rhuZP3 was 95%.Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA.Conclusion:The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.

Objective To investigate the inhibitory effect of 5-FU on human intrahepatic biliary epithelial cells with TGF-β1-induced mesenchymal transition and potential mechanism.Methods The primary epithelial cells from human intrahepatic bile ducts were cultured,the cells were identified by immunofluorescence microscopy.The cells were randomly divided into 3 groups: normal control group,TGF-β1 group and TGF-β1+5-FU group.The expression of CK-19,E-cadherin,vimentin and α-SMA protein were measured by immunofluorescence microscopy and Western blot assay.Western blot was used to detect the expression levels of TGF-β1.Real-time PCR to detect mRNA expression of TGF-β1,Ⅰand collagen type Ⅲ.Results The primary epithelial cells of intrahepatic bile ducts were successfully cultured.In TGF-β1 group,the protein expression of vimentin,α-SMA and the mRNA expression levels ofⅠand collagen type Ⅲ were significantly higher than that of normal control group(P<0.05),the protein expression of CK-19 and E-cadherin protein were lower than that of normal control group(P<0.05),TGF-β1protein and mRNA expressions were significantly higher compared with that of normal control group(P<0.05).In TGF-β1+5-FU group,the protein expression levels of CK-19 and E-cadherin protein were higher than that in normal control group(P<0.05),the protein expression level of vimentin,α-SMA and the mRNA expression level of collagen type Ⅲ were significantly lower than that of TGF-β1 group(P<0.05),while there was no significantly difference in collagen typeⅠmRNA between the two groups(P>0.05),TGF-β1 protein and mRNA expressions were significantly lower as compared with that of normal TGF-β1 group(P<0.05).Conclusions 5-FU could inhibit TGF-β1-induced biliary epithelial-mesenchymal transition with down-regulated expression of TGF-β1.

Objective To study genital toxicity and didymus lipid peroxidation level on rats after sub-acute exposure to di-2-ethylhexyl phthalate(DEHP).Methods The weaning Wistar andro-rats were dividied into four group randomly,and there were 10 rats in each group.Compared with the corn oil control,the experiment groups were gavaged consecutively for 30 days with the DEHP dose of 10,100,1 000 mg/(kg.d),The change of body weight was observed dynamically.All rats were executed by decapitation four weeks later and the didymium was removed immediately,at the same time viscera coefficient were measured.The level of MDA were detected by TBA method,meanwhile GSH level and GSH-Px activity were analyzed by modified DTNB method,the activity of SOD were measured by xanthine oxidase method.Results With the increasement of DEHP concentration,the body weight growth of rats in 1 000 mg/(kg.d) DEHP group is restricted compared with the control(P

Objective To research the protective effects of alkaline electrolytic water on lipid peroxidation in blood,liver,kidney and brain of mice. Methods Total forty-five Kunming mice were randomly divided into three groups,control group:tap water;peroxidation model group:100 mg/kg D-galactose subcutaneous injection (once a day) + tap water;experimental group:100 mg/kg D-galactose subcutaneous injection (once a day) + alkaline electrolytic water. After 18 weeks of exposure,the mice were sacrificed,and the blood,liver,kidney and brain of mice were collected for further study. The MDA levels were measured by TBA method; The GSH levels and GSH-Px activities were measured by modified DNTB method; The SOD activities were measured by Xanthine oxidase method; The lipofuscin levels were detrmined by Sohal method. Results Compared with the control group,MDA and lipofuscin level increased,GSH content decreased,the activities of GSH-Px and SOD decreased in all detected organs and blood in the peroxidation model group,and compared with the peroxidation model group,MDA and lipofuscin level decreased,GSH content increased,the activities of GSH-Px and SOD increased significantly in D-galactose +alkaline electrolytic water group. Conclusion Alkaline electrolytic water has some antagonistic effects on the lipid peroxidation induced by D-galactose.

Objective To investigate the role of indoleamine 2,3-dioxygenase(IDO)in asthma.Methods The mouse asthma model was induced by ovalbumin(OVA).IDO expression was detected on the level of protein and mRNA respectively.Distribution and maturation of dendritic cells were detected by the immunofluorescence method.Results (1)The symptoms and lung inflammation in the model group were more serious than control group.The serum total IgE was significantly higher in the model group than that in the control group,165.50 ± 30.13 ng/ml vs.94.45 ± 28.30 ng/ml(P ＜ 0.05).(2)IDO expression in the model group was lower than that in control group.On the level of protein,mean intergrated optical density was 11.38 ± 6.05 in the model group vs.23.62 ± 8.92 in the control group(P ＜ 0.05);on the level of mRNA,IDO expression of the model group was 33% of the control group(P ＜ 0.05).(3)CD11c+CD86+ cells were distributed in alveolar wall and around small vessels.The quantity of CD11c+CD86+ cells in lungs of the model group were significantly smaller than that in the control group.The median intergrated fluorescence intensity was 9961.86(range,7 406.52 ～ 12 724.98)in the model group vs.15974.60(range,10 006.39 ～ 16 171.46)in the control group(P ＜ 0.05).Conclusions IDO expression is low and matured dendritic cells are less in situ in the asthma model.These suggest that less matured DCs may produce less IDO,which may play an important role in asthma.

Objective The purpose of this study was to discuss the therapies for hemorrage caused by the fissuration of pancreatojejunal stoma and pancreatic leakage after pancreatoduodenectomy.Methods After three cases of pancreatoduodenectomy,the disruptions of pancreatojejunal stoma resulted in serious pancreatic leakage and the hemorrage in abdominal cavity.During all the second operations,the drainage-tube insertions into the main pancreatic ducts were used to lead the pancreatic juice into the neighboring loop of jejunum.Results Afer the operations,the supportive treatment,continuous irrigation of peritoneal cavity and pancreatic enzyme inhabition were given to the patients of these cases and all of the patients were successfully cured.Conclusions The bridge-crossing internal drainage which inserts drainage-tube into the main pancreatic duct was a convenient and effective therapy and method to rescue the hemorrage caused by the fissuration of pancreatojejunal stoma and pancreatic leakage after pancreatoduodenectomy.While the patients' lives were saved,their functions of pancreas were preserved and the qualities of life were improved after the operations.

Objective To evaluate the reliability of a rat model of acute cerebral ischemia and reperfusion by using cerebral perfusion functional CT.Methods A stable and reversible focal ischemia model with unilateral middle cerebral artery occlusion was established and evaluated by CT perfusion imaging and TTC staining.Results Artificial Occlusion of the MCA resulted in ipsilateral cerebral infarcts in all study animals.Hypoperfusion was definitely recorded in all CT perfusion images obtained after MCA occlusion and was significantly correlated with the final lesion size.Blood flow was restored after pulling the thread out of the artery.Conclusions The method of establishing an acute focal cerebral ischemia and reperfusion model by thread insertion in our study is simple and stable.If we can screen the stroke model with CT perfusion examination,the error caused by variance of model can be reduced.Thereby it provides a platform for researchers to investigate acute cerebral ischemia and recirculation.