Objective To discuss Foxp3 expression in peripheral blood from the patients with various stages of gastric cancer by SYBR Green I real-time PCR assay.Methods Foxp3 standard was prepared by conventional RT-PCR and employed to generate quantitative PCR standard curve.The precision,sensitivity and specificity of the assay were measured.SYBR Green I real-time PCR assay was adopted to detect and quantify Foxp3 expression in peripheral blood from 40 gastric cancer patients and 8 normal controls.Results The coding sequence (CDS) of full length Foxp3 was obtained and used as standards.The dynamic range of standard curve of real-time PCR assay was 102-108 copies.The mean coefficient of variation (CV) of inter-and intra-assay was less than 5%.The minimum detectable limit was 280 copies of Foxp3 mRNA.Foxp3 expression level in gastric cancer was significantly higher than that in normal control (P

Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.

Objective To investigate the populations of CD+4CD25high Foxp3+ regulatory T cells(Treg cells)and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer,and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages.Methods PBMC,Ascites,tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer(TNM I11,Ⅱ18,Ⅲ10,Ⅳ8)and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured witll a real-time quantitative PCR Results The percentage of CD4+CD25high Foxp3+ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthv donors(P<0.05).The percentage of Treg cells were more abundant in ascites(P< O.05),1rIL(P<0.01)and tumor-draining lymph nodes(P<0.01)of individuals with gastric cancer than in their blood.And the Foxp3 mean fluorescent intensity (MFI) of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites(P<0.05).The proportion 0f Treg cell in patients of gastric cancers with stage I,II,Ⅲ,Ⅳ were(5.72±1.95)%,(6.45± 2.23)%,(9.58±3.13)%,(11.70±2.30)%,respectively.There were significant correlation between Drevalenee of Treg cells and disease stages in gastric cancer(r=0.784,P<0.01).Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors.Foxp3 mRNA levels were correlated with TNM stages(r=0.623,P<0.05).Conclusions Our results suggest that the populations of CD4+CDhigh25 Foxp3+Treg cells and Foxp3 mRNA levels in patients with gastric cancer are signifieantlv higher in comparison to those in control donors.In addition,the expression of Fox：p3 mRNA increased with tumor stage.The increased prevalence of Treg cells may be one of the explanations for impaired cell-mediated immunity in cancer bearing hosts.

AIM: To investigate the relationship between up-regulation of plasminogen activator inhibitor-1 (PAI-1) expression and activation of p38 mitogen-activated protein kinase (p38 MAPK) and extracellular signal-regulated kinase ( ERK) pathways by TGF-β1 in human ovarian cancer cells .METHODS: PAI-1 expression in human ovarian cancer cells treated with TGF-β1 (10 μg/L)was assayed by real-time PCR and Western blotting.The activation of p38 MAPK and ERK was determined by Western blotting using phosphorylated p 38 MAPK and phosphorylated ERK antibodies . Specific p38 MAPK inhibitor (SB203580) or ERK inhibitor (PD98059) was used to inhibit their activation .RESULTS:TGF-β1 up-regulated the expression of PAI-1, and activated p38 MAPK and ERK pathways in the ovarian cancer cells .In-hibition of p38 MAPK activation by SB203580 resulted in significant inhibition of the mRNA expression of PAI-1 induced by TGF-β1.However, inhibition of ERK activation did not significantly alter TGF-β1-induced increase in PAI-1 mRNA level.CONCLUSION: TGF-β1-activated p38 MAPK pathway contributes to the up-regulation of PAI-1 expression by TGF-β1 in ovarian cancer cells .

Objective · To design and synthesize a series of benzenesulfonamide derivatives, test their inhibitory activity to CDK2-cyclinA2 kinase, and investigate the structure-activity relationship. Methods · Virtual screening was executed via computer-aided drug design according to the ATP binding site in CDK2-cyclinA2 protein crystal. A series of benzenesulfonamide derivatives were designed and synthesized on the basis of the interaction modes between the lead compound and the CDK2-cyclinA2. The biological evaluation of compounds was made through the CDK2-cyclinA2 in-vitro kinase activity detection system. Results · Twenty-nine new benzenesulfonamide compounds were prepared, and their inhibitory activity to CDK2-cyclinA2 was elicited. WZ-026 had the highest inhibitory parameter, which half maximal inhibitory concentration (IC50) was 3.81 μmol/L. Conclusion · By multipurpose utilization of virtual screening, chemical synthesis, and biological activity test, a benzenesulfonamide compound WZ-026 was found, which has great inhibitory activity towards CDK2-cyclinA2. Preliminary structure-activity relationship of compounds was obtained.

Objective To investigate the protective effect of ischemic preconditioning(IP) on liver ischemia/reperfusion(I/R)injury in rats with cirrhosis.Methods Thirty-two SD rats with liver cirrhosis induced by carbon tetrachloride were randomly divided into IP group and Non-IP(NIP) group,according to whether an IP was performed before liver I/R injury or not.The function of liver, superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),malonyldialdehyde(MDA),adenine nucleotide(ATP,ADP,AMP),energy charge(EC),nitric oxide(NO)in liver tissues,the changes in liver histology,and one-week survival rate after the test in both groups were compared.Results Compared with NIP group,the serum AST,ALT ,and LDH and the MDA contents significantly decreased(P

OBJECTIVE:To provide reference information for the adoptation of fast drug dispensing model in hospital pharmacy of China.METHODS:The procedure of fast drug dispensing system(FDDS)was analysed and compared with tradi?tional dispensing system in advantages and disadvantages.RESULTS:FDDS could raise the speed,accuracy of drug dispensing and improve the quality of pharmaceutical service.CONCLUSION:FDDS is suitable for drug dispensing in large-scale hospi?tals.

Objective To evaluate the distribution and drug resistance of bacteria causing neonatal infectious pneumonia in Jiaxing,and to provide a therapy for clinical doctor to make a correct diagnosis,choose reasonable anti-biotics and avoid abuse of antibiotics.Methods Took expectoration from trachea in condition of asepsis to conduct culture and perform drug-sensitive test from 3025 cases.Results Totally 1 156 strains of aerobic bacteria were iso-lated.875 strains were gram negative bacilli(75.7%),269 strains were gram positive cocci(23.3%),and 12 strains were fungi(1.0%).Klebsiella pneumoiae,Escherichia coil,Acinetobacter baumanni,Enterobacter cloacae were com-mon in gram negative bacilli( respectively 178 cases,151 cases,87 cases,113 cases) .The proportion of the Staphylo-coccus aureus was the largest in gram positive cocci(245 cases) .The results showed that gram-negative bacilli were resistant to cefazolin, ampicillin, piperacillin and sensitive to meropenem, imipenem, piperacillin-tazuobatanna and cefoperazone-sulbactam.The drug resistance was severe of ESBL-positive.Staphylococcus aureus was resistant to penicillin, ampicillin, erythrocin, clindamycin and sensitive to linezolid, vancomycin, nitrofurantoin.Conclusion Gram-negative bacilli are the main bacteria in neonatals with infectious pneumonia.The drug resistance is severe.It is important to make a standard management and isolation.

Objective To formulate the index framework of medium developed health level in accordance with Zhejiang Provinces development goal of primarily realizing modernization by 2020. Methods Such research methods as information retrieval, cluster analysis, and Delphi expert consultation were used to select index items of medium developed health level. Results 14 indexes were selected, falling respectively into 4 major categories, viz. health level of the residents, allocation of health resources, provision of and access to health services, and environment and form of the residents health and life. Conclusion The index framework is capable of reflecting in a comprehensive way the overall health level of a certain region.

Objective:To observe the effect of acupoint massage plus acupoint sticking therapy for the stress reaction during postoperative anesthesia recovery period in patients undergoing nasal endoscopic surgery.Methods:A total of 120 patients undergoing nasal endoscopic surgery were included,and all patients were under trachea intubation and general anesthesia.The patients were randomized into an observation group and a control group,with 60 patients in each group.Patients in the control group received conventional anesthesia resuscitation,while patients in the observation group received acupoint massage plus acupoint sticking therapy on the basis of conventional anesthesia resuscitation.Changes in the heart rate (HR),systolic blood pressure (SBP) and diastolic blood pressure (DBP) were observed at three time points including the end of the surgery (TO),the removal of the tracheal tube (T1) and 10 min after the removal of the tracheal tube (T2).The awakening and tube removal time,choking cough and restlessness,and adverse reactions (dizziness,nausea and vomiting) in 24 h post-surgery period were compared.Results:At T1 and T2,the comparisons of HR,SBP and DBP between the two groups showed statistical significance (all P＜0.05).Intra-group comparisons showed that the HR,SBP and DBP of the control group at T1 and T2 were significant different from those at TO (all P＜0.05).There were significant differences in the awakening time and tube removal time between the two groups (both P＜0.05).The incidences of choking cough and restlessness were 8.3％ and 3.3％ respectively in the observation group,versus 53.3％ and 30.0％ in the control group,and the between-group comparisons showed statistical significance (both P＜0.05).The incidences of dizziness,nausea and vomiting in 24 h post-surgery period were 3.3％,5.0％ and 0.0％ respectively in the observation group,versus 43.3％,33.3％ and 25.0％ in the control group,and the between-group comparisons showed statistical significance (all P＜0.05).Conclusion:Acupoint massage plus acupoint sticking therapy can effectively regulate the stress reaction during postoperative anesthesia recovery period in patients undergoing nasal endoscopic surgery,and maintain a stable internal environment.