A vector for a mutant Chinese Helicobacter pylori (Hp) strains knock out pathogenicity island (PAI) was constructed, which was the basis to establish a Chinese Hp mutant deleting pathogenicity island. Genetic engineering techniques such as polymerase chain reaction, plasmid extraction, agarose gel electrophoresis, restriction analysis, ligation, preparation of competence cell and transformation were used to make two cloning fragments containing two end regions of PAI and a selectable chloramphenicol resistance marker between them, and then engineering the recombined fragments into pBluescript plasmid. The result showed that restriction analysis demonstrated that the engineering mutant vector had been recombined. It suggested that we constructed an engineering mutant vector which targeted to delete the PAI in Chinese Hp strains. It will be useful for addressing the role of PAI in the pathogenesis of Hp infection.

0.05). The products of conjunction of cagⅠ and cagⅡ were found only in 5 strains. The detectable rate of continuous cag PAI was much higher in duodenal ulcer than in chronic gastritis ( P

Objective To explore the role of intercellular adhesion molecule-1(ICAM-1) in the metastasis of lung cancer.Methods The expression of ICAM-1 in the tissue and the serum ICAM-1(sICAM-1) level in the 52 cases of primary lung cancer were detected with immunohistochemical staining method(SP) and enzyme linked immunosorbent assay(ELISA),with 20 cases of adjacent non-cancerous tissue as the control.Results The positive rate of ICAM-1 expression in lung cancer was significantly higher than that of adjacent non-cancerous tissue.The expression of ICAM-1 in the cases with lymph node metastasis was higher than that of no lymph node metastasis.The expression of ICAM-1 in stage III+IV was higher than that of stage I+II,and that in adenocarcinoma was higher than that of squamous cell carcinoma.The level of sICAM-1 in patients with lymph node metastasis was higher than that without lymph node metastasis.The sICAM-1 level in patients of adenocarcinoma was higher than that of squamous cell carcinoma,and that in patients with positive expression of ICAM-1 in tissue was higher than that of negative expression of ICAM-1.Conclusion The expression of ICAM-1 may be associated with the metastasis in lung cancer.The sICAM-1 level reflects the expression of ICAM-1 in tissue and may be used as a tumor marker of metastasis and prognosis in lung cancer.

Objective To study the effect of postoperative ileus of laparoscopic surgery versus open surgery for colorectal canc-er .Methods 220 patients of colorectal cancer were selected from October 2009 to June 2012 .The patients were randomly divided into the observation group(n=110 cases) and the control group(n=110 cases) .The patients of the observation group were treated with laparoscopic surgery ,and the patients of the control group were treated with conventional open surgery .The postoperative anal exhaust time and hospital stay time ,the incidence of postoperative ileus of the two groups were observed and compared .Results The postoperative anal exhaust time and hospital stay time of the patients after radical resection of left colon ,right colon and rectal cancer of the observation group were significantly less than that of the control group (P<0 .05) .The incidence of postoperative ileus of the observation group was 5 .45% (6/110 ) ,significantly lower than 11 .82% (13/110 ) of the control group ( P< 0 .05 ) . Conclusion Laparoscopic colorectal surgery is a minimally invasive surgery ,the patient′s trauma is smaller ,the postoperative re-covery is rapidly .The incidence of intestinal obstruction is smaller than that of the conventional open surgery .

Objective To explore the effectiveness of 125I seed implantation for gastric cancer and to determine whether the therapy could increase the survival rate.Methods Seventy-six gastric cancer patients in stage Ⅱ or Ⅲ were involved and randomly divided into treatment group (n =42) and control group (n =34)by simple random sampling method.The patients in the control group underwent D2 or D3 surgery and the patients in treatment group underwent D2 or D3 surgery plus interstitial implantation of 125I seeds.All patients signed the informed consents.Treatment results were evaluated as CR,PR,NC and PD.CR and PR were considered as effective and the effective rate was calculated.All patients were followed up and the three-or five-year survival rate was calculated,the complications were examined.x2 test was used to compare the significant difference between the two groups.Results The total effective rate in control group was 50.00％ (17/34),lower than that of treatment group (73.81％,31/42; x2 =4.578,P＜0.05).In the treatment group,the three-year and five-year survival rates were 61.90％(26/42) and 42.86％(18/42) respectively,and the corresponding rates in the control group were 11.76％(4/34) and 0(0/34) respectively (x2=19.771,19.094,both P＜0.001).Both of the two groups had few severe side effects.Conclusion Radical surgery plus 1~Iseed implantation is effective and safe for the treatment of stage Ⅱ or Ⅲ gastric cancer and can further improvelong-term survival.

Objective To screen the specific antigen mimotopes of influenza A(H1N1)virus by phage display technology,in order to pave a way to develop novel influenza vaccine.Methods Using human convalescent serum of pandemic influenza A(H1N1)in 2009 as solid-phase selective molecule,an artificial synthesized phage randomly displaying cyclic 7-mer peptide was screened with three rounds of "absorption-elution-amplification" selection.At the end of the third round selection,32 clones were randomly chosen from the top-agar phage plaques and placed onto E.coli ER2378 in logarithmic growth phase.After culturing for 5 hours,the positive clones were identified by enzyme linked immunosorbent assay(ELISA),cross reaction test and competitive inhibition assay.The identified positive clones were analyzed by DNA sequencing.The decoded amino acids sequences,which displayed on the surface of phage,were aligned with the hemagglutinin(HA)gene of influenza virus by homology comparison for definition of the mimotopes of influenza A(H1N1)virus.Results After enriching the specific antibody-binding phages from phage displaying peptide library,12 positive clones were chosen from 32 randomly selected clones.DNA sequencing and homology comparison showed that one epitope PLHARLP was confirmed as mimotope of swine influenza A(H1N1)virus antigen,which was composed of the 52nd,53rd,59th,60th,61st,83rd and 271st amino acid.Conclusions Swine influenza A(H1N1)mimotope has been obtained by cyclic 7-mer peptide phage library screening.This result provides a basis for developing new influenza virus vaccine from virus antigen mimotopes.

Objective To investigate the relationship between serum ferritin and p-cell function,insulin resistance in elderly patients(age>60 years)with type 2 diabetes and impaired glucose tolerance.Methods Total 1143 patients with type 2 diabetes,448 patients with impaired glucose tolerance(IGT),and 2 950 subjects with normal glucose tolerance(NGT)were recruited for the measurments of height,weight,serum triglyceride(TG),total cholesterol,fasting plasma glucose(FPG),fasting insulin(FINS),postprandial 2 h plasma glucose(2hPG),and serum ferritin.Homeostasis model assessment of insulin resistance(HOMA-IR)and QUICK index were used to estimate insulin resistance,and homeostasis model assessment of β cell function(HOMA-p)and disposition index(DI)to evaluate p-cell function.Results The results showed that the level of serum ferritin was significantly higher in type2 diabetes mellitus group than NGT group(P<0.01).HOMA-IR was gradually increased,and HOMA-β,QUICK,and DI decreased from NGT to IGT,type 2 diabetes mellitus groups(all P < 0.05).Spearman and partial correlation analysis showed that partial adjustment for age and sex,serum ferritin was positively associated with FPG,2hPG,TG,FINS,and HOMA-IR,and negatively associated with HOMA-β,QUICK,and DI.Stepwise regression showed that serum ferritin was associated with QUICK,BMI,and TG.Conclusion The results suggest that iron overload exists in elderly patients with type 2 diabetes.With increasing serum ferritin level,insulin resistance increases and p-cell function decreases in the elderly patients with abnormal glucose metabolism.

Background and purpose: TFPI-2 is a new serine proteinase inhibitor,it is related to many tumors.In this study,the effect of TFPI-2 gene on cell proliferation and apoptosis of human pancreatic cancer cell line Panc-1 were investigated.Methods:The growth curve was drawn for the 3 groups,Panc-1-TFPI-2、Panc-1-V and Panc-1.DNA strand breaks were used to detect the apoptosis of the 3 groups,and the change of cell cycle and apoptosis index was evaluated by flow cytometry.Results:Compared with nontransfected cells,the growth of Panc-1 cell transfected with TFPI-2 was inhibited significantly.The transfected cells showed a significant increase in G1/G0 phase.The apoptosis of Panc-1-TFPI-2 cell could be identified by DNA strand breaks and flow cytometry in comparison with the control group.The apoptosis index of the Panc-1-TFPI-2 cell(6.9?0.5)% was higher than the group Panc-1-V and group Panc-1[(3.0?0.4)% and(3.5?0.4)%]P

Objective To investigate the invasion ability of Panc-1 cells in vivo and in vitro after being transfected with tissue factor pathway inhibitor 2 gene(TFPI-2).Methods The expression vector pEGFP-C1-TFPI-2 was transfected into human pancreatic cancer line Panc-1 cells by using liposome.TFPI-2 mRNA and protein of transfected and nontransfected cells were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot respectively.The tumor cells invasive behavior of transfected(Panc-1-TFPI-2)and nontransfected(Panc-1-V and Panc-1-P)cells were assessed in vitro through Boyden Chamber method.The transfected and nontransfected cells were implanted into nude mice to observe its growth and metastasis in vivo.Results Expressions of mRNA and protein of TFPI-2 were confirmed in transfected cells.After TFPI-2 transfection,the number of Panc-1-TFPI-2,Panc-1-V and Panc-1-P cells passing through membrane of Boyden Chamber were 24.4?3.5,61.3?4.1 and 60.2?3.9,respectively.The number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviously decreased compared with that of non-expressing cells,the invasion ability was lower than that before transfection in vitro.The subcutaneous tumor volume of the Panc-1-TFPI-2 group was(438.0?69.8)mm3,the Panc-1-V group was(852.0?102.9)mm3 and the Panc-1-P group was(831.0?78.1)mm3,P

Objective To explore the effect of long-term depleted uranium (DU)ingestion on testosterone production in rats, and its involvement mechanism. Methods Male and female rats (F0 and F1 respectively) for 160 days, respectively. The contents of testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) in serum were detected in 20 months of F0 generations, and 15 months of F1 generations. RT-PCR was used to analyze the levels of StAR mRNA and P450scc mRNA. Results Compared with the normal control group, the testosterone contents in exposed F0 and F1 generations increased, the lowest was 51.73 U/L, but those of LH and FSH decreased. The expression of StAR mRNA in the low-doze group of F1 generation (StAR/β-actin = 1.35) was up-regulated, down-regulated for other groups.compared with the normal control group (P450scc/β-actin = 0. 313), the expression of P450scc mRNA in the low- and high-dose groups of F0 generation were decreased (P450scc/β-actin = 0.21), and those in the low- and high-dose groups of F1generation were increased (P450scc/β-actin = 0.623) (P ≤ 0.01). Conclusion Long-term DU exposure inhibit the male reproduction by intervening the sexual hormone production through down-regulated the expression of StAR mRNA and P450scc roRNA.