Objective To analyze the seroprevalence of Helicobacter pylori (H. pylori) infection and its specific genes in liver tissues of chronic hepatitis B virus (HBV) infected patients, and to investigate the effect of H. pylori on development of chronic HBV infected liver diseases. Methods Five hundred and two patients infected with HBV and 429 sex-and age matched healthy controls were enrolled in the case-control study. All subjects were tested for presence of antibodies against H. pylori using ELISA. Fifty-six liver biopsy samples were amplified by polymerase chain reaction (PCR) using Helicobacter genus-specific 16S rRNA primers. The positive samples were further amplified using specific primers of H. pylori cagA, vacA and glmM genes. Results H. pylori infection was accounted for 63.9% in HBV infected patients, which was higher than that in healthy controls (43.4%,P<0.05). Moreover, the seroprevalence of H. pylori in patients with hepatocellular carcinoma (HCC, 29/36,80.6%) or cirrhosis (64/83,77.1%) was higher than that in patients with chronic hepatitis (228/383,59.5%, P<0.01). Helicobacter genus-specific 16S rRNA was found in 17,7 or 11 of patients with cirrhosis, HCC or chronic hepatitis. Twenty-one samples were confirmed as H. pylori DNA by PCR. Conclusions The seroprevalence of antibody against H. pylori was higherHelicobacter can be detected in liver tissues of HBV infected patients. H. pylori might play the role in the development in HBV infected patients compared with healthy controls. Besides H. pylori, other of chronic hepatitis to cirrhosis and HCC.

Objective To research the role of Helicobacter pylori(Hp) on chronic hepatitis B.Methods The seroprevalence of Hp infection and the quantity and genotyping of HBV DNA in 376 patients with chronic hepatitis B,including chronichepatitis group,cirrhosis group and HBV-related hepatocellular carcinoma(HCC) group,were detected,and compared with control and gastritis groups.Results Hp seropositivities in chronic hepatitis B group(56.2%),cirrhosis group(69.9%),HCC group(75.0%) were higher than that in control group(43.4%)(P0.05),the Hp seropositivities in cirrhosis and HCC groups were higher than that in chronic hepatitis group(P0.05).Conclusion Seroprevalence of antibodies to Hp in patients with chronic hepatitis B increases significantly,and Hp seropositivity increases with the pathological changes of chronic hepatitis B.

0.05).Conclusion Among the DM patients,the incidence rate of PC is obviously increasing,and with the course of DM elongating,the risk of PC is increasing.There is no obvious difference between sexes.Compared with common people,the incidence rate of PC in DM patients is higher.Type 2 diabetes mellitus might be initial symptom of PC.

Objective To compare the Helicobacter pylori (Hp) eradication rate of different therapies and to explore the effects of Hp eradication on the clinical characteristics of chronic obstructive pulmonary disease (COPD).Methods From December 2006 to December 2009,at China-Japan Union Hospital of Jilin University 89 stable COPD patients with Hp infection were divided into eradication group and non-eradication group.The eradication group was divided into clarithromycin sub group and moxifloxacin sub group.The patients of these three groups all received regular COPD treatment.Esomeprazole,amoxicillin,clarithromycin and colloidal bismuth citrate were used in clarithromycin group.Esomeprazole,amoxicillin,moxifloxacin and colloidal bismuth citrate were used in moxifloxacin sub group.Patients received pulmonary function test,exercise tolerance evaluation,dyspnea scoring and health-related quality of life scoring at recruitment and 12 months after recruitment.The onset frequenly of acute exacerbation of COPD in one year was counted.The data were analyzed by x2 test and t test.Results The Hp eradication rate of clarithromycin sub group (48.4 ％,15/31) was lower than that of moxifloxacin sub group (87.1％,27/31),and the difference was statistically significant (x2 =4.22,P=0.032).There was no significant difference percentage of forced expiratory volume in first second to forced vital capacity in (FEV1％) predicted value between 27 cases in non-eradication group and 53 patients with successful Hp eradication (t=0.677,P=0.265).Of 53 patients with successful Hp eradication,the 6-min walking distance,Borg dyspnea score and saint George's respiratory questionnaire (SGRQ) score were improved significantly (t =1.884,1.877 and 1.773 respectively; P=0.032,0.025 and 0.034 respectively),and there was no improvement in 27 non-eradication patients.There was significant difference in the frequency of COPD acute attack between 53 patients with successful Hp eradication (1.2 times) and non-eradication group (1.9 times) (t=1.812,P =0.034).Conclusions Hp eradication therapy with moxifloxacin in COPD patients reached higher Hp eradication rate.Hp eradication in COPD patients with Hp infection can improve the exercise tolerance of patients,relieve dyspnea,improve quality of life and reduce the frenquency of acute attacks.

Objective To investigate the prevalence of Helicobacter hepaticus (H. hepaticus)infection in various species of mice from different regions of China in order to find the role of H. hepaticus in development of hepatitis, liver cancer and tumors in lower digestive tract in mice.Methods One hundred and fourteen mice, including C57BL/6 mice (n= 39), BABL/C mice (n=45),SCID mice (n=14) and C3H mice (n=18), were collected from different regions of China. The serum anti-H, hepaticus-IgG and fecal H. hepaticus antigen were determined by using ELISA. Polymerase chain reaction analysis (PCR) was used to screen Helicobacter genus-specific 16SrRNA and H. hepaticus species-specific 16SrRNA. The feces were cultured and identitied for Helicobacter infection in 114 mice. The H. he paticus infection was identified as one of above tests being positive.Results Of 114 mice, 25 (21.9%) mice were infected with Helicobacter species. The mice infected with H. hepaticus accounted for 44. 0% (11/25) with SCID and C3H mice in high prevelence.Meanwhile, the PCR examination revealed that the rest 56.0% (14/25) mice were infected with other Helicobacter species. Conclusion Besides H. hepaticus infection, the other Helicobacter species infections are also existed in China.

BACKGROUND: The cryopreservation of human ovarian tissue has become an attractive method to preserve female fertility. Human ovarian tissue experiences neovasculadzation after transplantation to recover blood supply, cryopreservation and resuscitation technique is a key for the neovascularization of human ovarian tissue following transplantation. OBJECTIVE: To investigate vascular endothelial growth factor (VEGF) expression and microvessel density in human ovadan tissue following novel needle immersed vitrification (NIV) and slow-freezing, to explore the influence of two cryopreservation methods play in the neovascularization of human ovarian tissue after transplantation. METHODS: Eight normal human ovarian tissues from patients with carcinoma of endometrium were cut into 12 fragments in the size of 1.5 mm × 1.5 mm × 1.0 mm, then randomly assigned to 3 groups: fresh control group, NIV group and slow-freezing group. In the NIV group, pieces of ovarian tissue stdps were dehydrated in an equilibration solution consisting of 7.5% ethylene glycol and 7.5% dimethyl sulfoxide in TCM-199 supplement with 20% fetal bovine serum and a vitrification solution consisting of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 mol/Lsucrose, then were plunged in liquid nitrogen directly and sealed in liquid nitrogen-filled cryovials. For thawing, the needles holding ovadan tissues were serially transferred into 1.0, 0.5, 0.25 mol/L sucrose solution and TCM-199 supplemented with 20% fetal bovine serum. In the slow-freezing group, pieces of human ovadan cortex fragments were placed in a 1.8-mL cryovial containing 1 mL of TCM-199 medium supplemented with 0.1 mol/L sucrose, 20% fetal bovine serum and 1.5 mol/L dimethyl sulfoxide, the cryovials were placed in the programmable freezer and cryopreserved by preset slow-cooling protocol. For thawing, the ovarian tissue stdps were washed in a stepwise manner: 1.0 mol/L dimethyl sulfoxida + 0.1 mol/L sucrose, 0.5 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose, 0.25 mol/L dimethyl sulfoxJde + 0.1 mol/L sucrose and 0.1 mol/L sucrose. The frozen-thawed and fresh controlled human ovarian tissues were cultured in vitro. The expression of VEGF and CD34, as well as microvessel density, was analyzed by immunohistochemistry. RESULTS AND CONCLUSION: There was patchy and mild expression of VEGF in the stromal cells of all the three groups before and after culture. The expression of VEGF increased and reached peak value after culture for 2 days, began to decrease after culture for 4 days and further attenuated after culture for 6 days in all the three groups. Compared with slow-freezing group, the expression of VEGF in NIV group was closer to that in fresh control group. Microvessel density of all the three groups increased and reached peak value after culture for 2 days, and the microvessel density of fresh control group and NIV group was significantly higher than that of slow-freezing group (P < 0.05). The microvessel density of slow-freezing group after culture for 4 days and that of all the three groups after culture for 6 days significantly decreased compared with after culture for 2 days (P <0.05). NIV is superior to slow-freezing to preserve stromal cells and extracellular matrix of human ovarian tissue, and plays less influence in VEGF expression and angiogenesis in human ovarian tissue.

Objective To study the transfeetion efficiency and safety of liposome microbubble(LM)on red fluorescent protein(RFP)in vitro and in vivo under ultrasound mediated gene transfection(USMGT)conditions.Methods Plasmids containing RFP were added to cultured Hela cells followed by ultrasound (US)exposure with LM.Different concentration of LM,US intensity and exposure time were optimized.Transfection efficiency was evaluated by fluorescent microscopy and FACS.Cell viability was verified by propidium iodide assay.In transplanted tumors in vivo study,LM and plasmid(P)were injected into the nude mice followed by US exposure(P+LM+US group).Nude mice undergoing plasmid injection alone(P group),plasmid injection and US exposure(P+US group)and plasmid and LM injection(P+LM group)were used as controls.Frozen section and histological examination were conducted and RFP expression was evaluated.Results LM and US exposure significantly increased transfeetion efficiency in cultured Hela cells (P＜ 0.01).Transfection efficiency was the most prominent under the condition of US intensity of 1.0 W/cm2 with 6%LM,duration 3 min.No apparent cell damage was found in the all groups.In transplanted tumors,strong RFP was seen in P+LM+US group.It was significantly higher than in any other groups(P＜0.0 1).No tissue damage was seen histologically.Conclusions LM could enhance USMGT effectively without causing any apparently adverse effect in vitro and in vivo.This method would be a novel,effective,safe non-viral gene transfection method and provide an alternative to current clinical gene therapy.

Objective To investigate the prevalence of Helicobacter pylori (H .pylori)infection in type 2 diabetic patients and its effects on diabetic gastroparesis.Methods Prospective clinical case-control study was applied.From January 2011 to December 2013,125 hospitalized patients with type 2 diabetes and 142 healthy controls without dyspeptic symptoms were enrolled.The prevalence of H .pylori infection and the incidence of gastroparesis in 125 patients with diabetes were investigated in both two groups.The patients with type 2 diabetes were divided into groups according to the course of the disease,and the prevalence of gastroparesis and H .pylori infection of each group were analyzed.The patients with type 2 diabetes and healthy controls confirmed with H .pylori infection were treated with eradication therapy,the rate of eradication of two groups was compared.The improved symptoms of gastroparesis before and after eradication therapy of patients with type 2 diabetes were compared.The chi-square test was performed for statistical analysis.Results The prevalence of H .pylori infection in type 2 diabetic patients was 66.4%(83/125),which was significantly higher than that of healthy control group (51 .4%,73/142 )(χ2 =5 .549,P <0.05).The prevalence of gastroparesis in diabetic patients with the disease course less than 10 years,10 to 20 years and more than 20 years was 33.8% (27/80 ),47.1 % (16/34 )and 8/11 , respectively.The difference was statistically significant (χ2 = 6.554,P < 0.05).The prevalence of H .pylori infection in patients with gastroparesis was 78.4% (40/51 ),which was significantly higher than that of patients without gastroparesis (58.1 %,43/74)(χ2 =4.716,P <0.05).The eradication rate of H .pylori infection in patients with type 2 diabetes was 68.7% (57/83),which was lower than that of healthy control group (87.8%,36/41),and the difference was statistically significant (χ2 =4.385 ,P <0.05).The incidence of epigastric pain and distension,early satiety and apocleisis before H .pylori eradication in type 2 diabetes patients was 75 .9% (63/83 ),66.3% (55/83 )and 67.5 % (56/83 ), respectively,while after eradication which was 44.6%(37/83),37.3%(31/83)and 39.8%(33/83)after eradication,respectively.The differences were statistically significant (χ2 =15 .720,12.764 and 11 .724;all P <0.01).Conclusions The prevalence of H .pylori infection is significantly higher in type 2 diabetic patients,and gastroparesis in type 2 diabetic patients may be correlated with H .pylori infection.The eradication rate in type 2 diabetic patients was lower,and H .pylori eradication therapy can efficiently improve the symptoms of dyspepsia in diabetic patients with gastroparesis.

Objective To establish Helicobacter pylori (Hp) infection induced chronic obstructive pulmonary disease (COPD) rat model,and to explore the role of Hp in the pathogenesis of COPD.Methods40 Wistar rats were randomly divided into double modeling group (Hp infection,smoked and intratracheal instillation of lipopolysaccharide),COPD group (smoked and intratracheal instillation of lipopolysaccharide),Hp infected group and control group.The lung function,cytokines level in serum and bronchial alveolar lavage fluid (BALF),Hp related genes expression in bronchial and lung tissue were detected.And Hp in bronchial and lung tissue was isolated and cultured.Results The lung tissue of both COPD group and double modeling group accorded with COPD pathological characteristics,and the latter was more apparent.The lung function of COPD group and double modeling group decreased more significantly than that of control group and Hp infected group (all P＜0.05),and which was more obvious in double modeling group than that of COPD group (P＜0.05).Along with the Hp colonization density increased,Ri and Re value of double modeling group increased (r=0.785 and 0.905),the value of Gdyn,PEF and FEV0.3/FVC decreased (r=-0.975,-0.959and -0.976).Compared with control group,IL-6,IL-8 and TNF-a cytokines levels in serum and bronchoalveolar lavage fluid of other groups increased significantly (all P＜0.05),and within the groups,double modeling group increased most significantly (all P＜0.05).Hp UreC gene was only amplified in part of bronchi and lung tissue of double modeling group,no Hp and suspicious bacteria colonies were isolated and cultured.ConclusionsHp not directly colonized in bronchi and lung tissue,which aggravated inflammation through increasing the serum and bronchoalveolar cytokines level of COPD rat model.Which caused the deterioration in lung function of COPD group.

Objective To investigate the value of time parameters(the transit time in coronary and the half filling time in myocardium)in assessing the blood flow of ischemia myocardium through establishing canine model of acute myocardial infarction.Methods Eighteen healthy anesthetized open-chest dogs were ligated the left anterior descending(LAD),3 hours later the contrast agent(C3F8)was injected into femoral vein in a fixed velocity to perform myocardial contrast echocardiography(MCE)examination.The first perfusion and the replenishment processes were recorded by real-time tri-plane MCE(RT-TP-MCE)and real-time single-plane MCE(RT-SP-MCE)respectively.both the two different processes were analyzed by the function Y=A ×(1-e-βt)+B.RT-SP-MCE used A(plat of peak intensity)and β(slope rate of the curve)values to quantitatively calculate the myocardial blood flow(MBF=A×β).The time point of the first microbubbles coming into the left ventricle was normalized to O in RT-TP-MCE,the time point of the first microbubbles coming into the myocardium was defined as the transit time in coronary(Tc),and the time point when microbubbles in myocardium achieved the 50% plat video intensity was defined as the half filling time in myocardium(Tm).Evans blue dye and TTC staining were performed to identify normal.ischemic and infarct myocardium.Results The time parameters of the two groups-normal myocardium and ischemic myocardium from RT-TP-MCE were:Tc(10.1±1.3)s and(20.4±7.1)s(P＜0.01),Tm(17.1±2.2)s and(39.7±8.8)s(P＜0.01).There were significantly negative correlation between the time parameters(Tc,Tm)and MBF from RT-SP-MCE in ischemic myocardium(r=-0.876,P＜0.01;r2=-0.894,P＜0.01).Conclusions The first perfusion imaging with RT-TP-MCE could be used to simultaneously,quantitatively evaluate the myocardial perfusion.The lower flow,the longer transit time in coronary and half filling time in ischemic myocardium.