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 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 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 the causes and management of the reoperation after cholangiointestiostomy(CIS). Methods　A retrospective analysis was made on the clinical data of 28 cases of reoperation after CIS from June 1995 to June 1999. Results　Among the 28 cases, 26 cases(92%) had CIS anastomotic stenosis. Of the 26 cases, 9 cases accompanied with left hepatobiliary duct stenosis, 3 cases with right hepatobiliary duct stenosis, 5 cases with left and right hepatobiliary ducts stenosis. 9 cases with biliary reflux comfirmed by barium meal radiography, all of the 9 cases were subjected to a choleduodenostomy. Of the 28 patients, 3 underwent reanastomose after excision the primary anstomosis, 8 operated with hilar bile duct reform and left lateral hepatolobectomy, 2 with left hepatic duct jejunostomy and hilar bile duct jejunostomy. 15 cases with intrahepatic bile duct jejunal Roux-en-Y anastomosis, after resolved the intrahepatic bile duct stenosis. Conclusions　The basic cause of reoperation after CIS is anastomotic stenosis, the other causes are as follows: the selected operation is unsuitable, the intrahepatic bile duct stenosis is not resolved, and the stonedoes not clean out completly. When reoperation is performed on these cases, the following principles must be abided by: romoving all the stones, resolving the stenosis, making a clear drainage; and performing hepatic lobectomy, anastomotic sustaining and drainage, and cholefibroscopic management must be done if needed.

Objective To study the regulatory role of chemerin in infant with respiratory syncytial virus (RSV) pneumonia by investigating the level of serum chemerin,pro-inlfammatory cytokine (TNF-α, IL-17), and anti-inlfammatory cytokine (IL-10, TGF-β). Methods The serum level of chemerin,TNF-α,IL-1,IL-10,TGF-βwere tested in 82 RSV pneumonia inpatients (17 severe RSV pneumonia cases,65 mild cases) and 40 controls by ELISA and the severity of the RSV pneumonia was evaluated using a scoring system. Results The serum level of chemerin of RSV pneumonia cases were (610.45±106.63pg/ml) which were signiifcantly higher than the control(337.24±43.37 pg/ml). Chemerin level of severe RSV pneumonia group is signiifcantly higher than mild cases as well [(786.62±82.59 pg/ml)vs (539.98±65.86 pg/ml)P<0.01 ]. Signiifcant positive correlations were found be-tween serum chemerin level and TNF-α,IL-17 level (r=0.81,r=0.61;P<0.01) while the serum level of chemerin is negatively correlated with IL-10, TGF-β(r=-0.80,r=-0.75;P<0.01). Conclusions The level of chemerin increased in RSV pneumonia patients,and related to clinical severity after RSV infection. These results indicate that chemerin may play an important role in the pathogenesis of RSV pneumonia and to the severity of the infection.

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.

Objective:To explore the influencing factors of false-positive serological reaction of syphilis.Methods:A total of 166 patients with false-positive serological reaction of syphilis (false-positive group), 145 patients diagnosed with early syphilis without treatment (positive control group) and 124 persons undergoing entry physical examination (negative control group) were included from January 2017 to February 2020 in Beijing Tongren Hospital, Capital Medical University. The gender, age and underlying diseases of the three groups were compared. Logistic regression was used to analyze the influencing factors of false-positive serological reaction of syphilis. The efficacies of chemiluminescence immunoassay (CLIA) and toluidine red unheated serum test (TRUST) were compared. Paired t test or chi-square test was used for statistical analysis. Results:In the false-positive group (166 cases), the age of 117 cases were more than 50 years old and 49 cases <50 years old. There were significant differences in age ((53.1±13.8) vs (24.7±2.8), t=22.56, P<0.01), autoimmune disease (36.7%(61/166) vs 6.5%(8/124), χ2=35.93, P<0.01), hepatitis (9.6%(16/166) vs 3.2%(4/124), χ2=4.92, P=0.026) and tumor (6.6%(11/166) vs 0.8%(1/124), χ2=4.68, P=0.030) between the false-positive group and the negative control group. There were significant differences in gender (there were 91(54.8%) males and 75(45.2%) females in the false-positive group, and 103(71.0%) males and 42(29.0%) females in the positive control group, χ2=8.67, P=0.003), age ((53.1±13.8) vs (34.4±12.9), t=20.13, P<0.01) and autoimmune disease (36.7%(61/166) vs 6.9%(10/145), χ2=39.14, P<0.01) between the false-positive group and the positive control group. Multivariate logistic regression analysis showed that gender (odds ratio ( OR)=2.692, 95% confidence interval ( CI) 1.504-4.816, P=0.001), age ≥50 years old ( OR=30.512, 95% CI 15.959-58.335, P<0.01), autoimmune disease ( OR=2.677, 95% CI 1.258-5.695, P=0.011) and hepatitis ( OR=4.408, 95% CI 1.799-10.799, P=0.001) were the influencing factors of false-positive serological reaction of syphilis. In the false-positive group, the positive rate of TRUST was 84.9% (141/166), which was higher than that of CLIA (23.5%(39/166)). The difference was statistically significant ( χ2=126.25, P<0.01). CLIA was 1.0-10.0 cut off index (COI) in 36 patients, and >10.0 COI in three patients.The difference was statistically significant ( χ2=52.51, P<0.01). The titers were ≤1∶4 in 139 patients and≥1∶8 in two patients with TRUST positive.The difference was statistically significant ( χ2=262.35, P<0.01). The sensitivity and specificity of CLIA were 95.2% and 96.0%, respectively, and those of TRUST were 77.2% and 91.1%, respectively. Conclusions:The influencing factors of false-positive serological reaction of syphilis include patients age ≥50 years and with autoimmune disease or hepatitis. The false-positive rate of TRUST is significantly higher than CLIA.