In the present study, the expression, purification, toxicity detection and the adjuvant effect of the heat-labile enterotoxin in non-toxic mutant mLT63 of Escherichia coli were investigated, in which the inductive expression was performed under optimal condition for inductive expression and the toxicity of the products obtained from inductive expression were tested for toxicity after being purified and concentrated with affinity chromatography. BALB/c mice were immunized orally with the mutant mLT63 associated with Helicobacter pylori (Hp) subunit vaccine UreB, Omp11. After immunization, the specific antibody levels in serum, extract from gastric tissues and fecal extract were determind by means of ELISA assay and the results were subjected to statistical analysis. It was demonstrated that the mutant mLT63 of heat-labile entrotoxin of E.coli constructed in our laboratory devoided of any toxic effect as revealed by the rabbit ileal loop assay, but its adjuvant effect could be demonstrated in the associated immunization of mice with Hp subunit vaccine UreB and Omp11.

Objective To investigate the effect of hypoxia-inducible factor-1α expression (HIF-1α) on toll-like receptor 4 (TLR4) signaling pathway-mediated rat lung ischemia-reperfusion injury (LIRI).Method Forty-five S-D rats were randomly divided into Sham group,LIRI group,LIRI+ TLR4-activated group,LIRI+ TLR4-inhibited group,LIRI + ASK1-inhibited group,LIRI + p38-inhibited group,and LIRI + HIF-1α-inhibited group.The interaction between TLR4 signaling pathway [including TLR4,myeloid differentiation factor 88 (MyD88),TIR-domain-containing adapter-inducing interferon-βTIR-domain-containing adapter-inducing interferon-β (TRIF),Apoptosis signal-regulating kinase 1 (ASK1) and p38] and HIF-1α and the role of TLR4-dependent HIF-1α in LIRI in vivo were analyzed.Result In LIRI,HIF-1α accumulation was induced in a TLR4-dependent fashion,and MyD88,but not TRIF,and activation of ASK1 and P38 were found to be critical for TLR4-mediated HIF-1α accumulation.HIF-1α protein played a critical role in TLR4-mediated lung injury of LIRI.HIF-1α up-regulated TLR4 expression in LIRI in a positive feedback manner.Conclusion We identify that HIF-1α has a damaging effect on TLR4 signaling pathway-mediated LIRI and TLR4-HIF-1 may synergistically involved in the development of LIRI.Therefore we suggest that the interaction between them may represent a novel therapeutic target for the development of novel target-based therapies of LIRI.

Objective To analyze the flavonoid content of common fruits consumed in China.Method Thirty-eight fruits were sampled from Beijing,Tianjin,Shanghai and Hangzhou.The contents of quercetin,kaempferol,luteolin,myricetin,apigenin were determined with high-performance liquid chromatography.Results Pomegranate and hawthorn were the highest in contents of 5 flavonoids among 38 fruits.Quercetin distributed more widely and was more abundant than other flavonoids in fruits.The daily intake of 5 flavonoids from fruits by Chinese people was estimated to be 2.80 mg.Conclusion The content of flavonoids varies significantly among different fruits and quercetin is the most common flavonoid contained in fruits.

Objective To analyze the expression and clinical significance of tumor suppressor phosphatase -tension protein gene(PTEN)in differentiated thyroid carcinoma and the relationship with the V600E gene site muta-tion of murine sarcoma viral oncogene homolog B1(BRAF).Methods The expression of PTEN was assayed by im-munohistochemical staining in differentiated thyroid carcinoma and adjacent tissues.The differences were compared between clinical pathological features.The BRAF V600E gene mutation was detected by fluorescence quantitative pol-ymerase chain reaction(PCR)and to investigate the correlation between the expression of PTEN and BRAF V600E mutation.Results The negative rate of PTEN was 78.75%(63 /80)in differentiated thyroid carcinoma,13.75%(11 /80)in adjacent tissues(χ2 =27.236,P =0.000).There were significant differences in the expression of PTEN between I -II and III -IV in TNMstage,grade I and II,and lymph node metastasis(χ2 =10.395,6.948,9.263,P =0.000,0.006,0.000).The differences were not significant in the expression of PTEN between different gender,≤45 years and >45 years different pathological types and tumor diameter <2cm and ≥2cm(χ2 =1.113,0.941, 2.301,1.567,P =0.185,0.213,0.087,0.181).There was correlation between the expression of PTEN and BRAF V600E mutations(r =0.301,P =0.004).Conclusion The expression of PTEN is involved in the occurrence and development of differentiated thyroid cancer and is associated with BRAF V600E.The PTEN has the predictive value of prognosis in tumors.

Objective:To construct an E.coli expressing system of human interferon-?(IFN-?).Methods:Extracted DNA from human blood and PCR human IFN-?, cloned the human IFN-? gene into plasmid T-easy and pBV-220. Expressed human IFN-? in E.coli DH5?, the expressing product was analysed by SDS-PAGE, Western blot and anti-virus capacity test.Results:DNA sequence analysis showed the recombinant plasmid pBV- IFN-? contained human IFN-?. SDS-PAGE and Western blot proved that there were hIFN-? in E.coli DH5? after temperature inducing and the expressing product has anti-virus activity.Conclusion:A human IFN-? E.coli expressing system was constructed successfully, and the recombination human IFN-? has anti-virus activity.

Objective To analyze the flavonoid content of common vegetables consumed in China.Method Forty-six vegetables were sampled from Beijing,Tianjin,Shanghai and Hangzhou.The contents of quercetin,kaempferol,luteolin,myricetin,apigenin were determined with high-performance liquid chromatography.Results Mei celery sampled from Hangzhou was the highest in content of 5 flavonoids among 46 vegetables.Quercetin distributed more widely and was more abundant than other flavonoids in vegetables.The daily intake of 5 flavonoids from vegetables by Chinese people was estimated to be 13.90 mg,in which quercetin accounted for 51.1%.Conclusion The content of flavonoids varies significantly among different vegetables and quercetin is the most common flavonoid contained in vegetables.The dietary intake of flavonoids was more from vegetables than fruits in Chinese people.

AIM: To investigate the antagonistic effect of zinc on hepatic ischemia- reperfusion (HIR) injury and the effect of zinc on the expression of adhesion molecules in rat liver to clarify the mechanisms involved. METHODS: After zinc supplementation (5 ?mol/kg bw,po) , the changes in hepatocellular morphology,ALT and AST activities in serum,MDA levels in serum,and expression of ICAM-1,VCAM-1 in liver of the animals subjected to HIR were examined. RESULTS: The results showed that HIR (30 minutes of ischemia followed by 90 minutes of reperfusion) significantly increased ALT, AST activities and MDA levels in serum. The destruction of hepatic structure was observed in HIR rats. In the mean time, the expression of adhesion molecule was enhanced.After zinc administration, ALT and AST activities in serum and MDA levels in serum were all decreased. The structure of hepatocyte was nearly normal.The further experiment showed that adhesion molecule expression was suppressed.CONCLUSION: These results indicate that zinc may protect liver against ischemia-reperfusion injury by inhibiting the production of free radicals and the expression of adhesion molecule.

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome. Methods PCR-RFLP was used to detect lipoprotein lipase Hind Ⅲ and S447X genotypes in 401 subjects(including 201 controls, 200 metabolic syndrome patients). Results ( 1 ) The levels of waist circumference ( WC ) , hip circumference ( HC ) , waist-to-hip ratio ( WHR ) , body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure ( DBP) , total cholesterol ( TC) , triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and fasting plasma glucose (FPG) were significantly different between metabolic syndrome group and control group (all P< 0.05). (2)The frequencies of H+H+ genotype,H+allele,SS genotype, and S allele for metabolic syndrome were all significantly higher than those for controls( H+H+ genotype:66. 5% vs 54.2% ,P=0.012; H+ allele:78.0% vs 71.4%, P=0.031;SS genotype:89.5% vs 77. 1% , P = 0.001; Sallel:94.5% vs 87. 56% , P = 0.001). (3) The levels of WC, HC, WHR, BMI, SBP, DBP, TG, LDL-C, and FPG in H + H-/H-H- genotype were significantly lower than those in H+H+ genotype, HDL-C was significantly higher than that in H+H+ genotype ( all P<0. 05). The levels of WC, HC, WHR, BMI, SBP, DBP, TC, TG, and FPG in SX/XX genotype were significantly lower than those in SS genotype, HDL-C was significantly higher than that in SS genotype ( all P< 0.05). (4)Multi-way logistic regression analysis suggested that risk factors for metabolic syndrome were smoking, drinking, and SS genotype (OR value was 4.289,2.268, and 2. 597, respectively ). (5) Result of interaction analysis among different factors indicated that the risk for metabolic syndrome in smoker with SS genotype was 3. 996 times of non-smokers with SX/XX genotype. Conclusions The lipoprotein lipase gene S447X polymorphism is associated with metabolic syndrome risk in Kazakh, and SS genotype and S allele may serve as genetic risk factors of metabolic syndrome, H + H-/H-H- and SX/XX genotypes yield beneficial effect for lipid and blood pressure. SS genotype and smoking may exist additive effect.

Questionnaire-based survey, physical examination, and blood testing were conducted according to cluster random samplings in Kazakh residents in Xinjiang.2 760 samples were collected to analyze the association of different strata of waist circumference and clustering of metabolic syndrome (MS) components.Accoding to International Diabetes Federation standard, the prevalence of ≥1and ≥2 components of MS showed increasing trend with the increase of waist circunference, and odds ratio of clustering of MS components also increased significantly.The distance of receiver operating characteristic curve was the shortest and the prevalence of MS was 22.1% ;22.4% in men, and 21.9% in women;when the waist circumference was ≥91 cm for men, and ≥88 cm for women.