Objective To analyze the influence of Helicobacter pylori (H.pylori) and smoking on serum pepsinogen (PG) levels in gastric adenocarcinoma (GAC).Methods Serum PGⅠand PGⅡ levels were measured by using radioimmunoassay method;H.pylori infection was determined by Hp-IgG antibodies in stored serum samples by using enzyme-linked immunosorbent assay and 14C urea breath test.Results In H.pylori negative cases,the serum PGⅠmean level in current smoking GACs was significantly increased compared with those non-smoking or ever-smoking GACs (the former:t=2.709,P

Objective To investigate the effect of recombinant human endostatin combined with chemotherapy on the expression of vascular endothelial growth factor (VEGF) in patients with advanced cancer.Methods250 patients with advanced malignant tumors were randomly selected from February 2012 to February 2016 the Third Hospital of Jiaxing, with unit control method experiment principle divided into two groups: experimental group (125 cases) and control group (125 cases).The experimental group treated by recombinant human endostatin combined chemotherapy, control group received routine chemotherapy for a week during the period of 21 days, after two cycles of treatment, clinical efficacy and adverse reactions of the two groups of patients were compared, and the expression of VEGF in tumor blood vessels weredetected.ResultsAfter two cycles of treatment, objective response rate, disease control rate in the experimental group patients was significantly higher than that in control group (P<0.05);after treatment, the expression level of VEGF in serum of the experimental group and the control groupwere decreased significantly, but the experimental group was lower than that of control group was more obvious (P<0.05).ConclusionRecombinant human endostatin combined with chemotherapy in the treatment of advanced malignant tumors can enhance the effect of chemotherapy, regulate the level of VEGF.

Objective To observe the influence of calcitriol combined with prolonged hemodialysis on parathyroid hormone (PTH),phosphorus and calcium in maintenance hemodialysis patients with refractory secondary hyperparathyroidism.Methods Twenty-two maintenance hemodialysis patients received prolonged hemodialysis (3 times per week and 6 hours each session) and therapy with calcitriol (3 times per week and 1 μ g each time) for 3 months.The levels of serum PTH,calcium and phosphorus of these patients were recorded before and after treatment.Results Compared with that before treatment,the level of serum PTH and phosphorus was decreased significantly[(484.21 ± 230.18) nmol/L vs.(750.53 ± 327.41) nmol/L,(1.49 ±0.27) mmol/L vs.(2.37 ±0.76) mmol/L],but calcium was increased after treatment [(2.35 ±0.32) mmol/L vs.(1.81 ±0.53) mmol/L],and there was significant difference (P ＜0.05).Conclusions The treatment of calcitriol combined with prolonged hemodialysis not only effectively clear phosphorus and PTH but also increase calcium in maintenance hemodialysis patients.The therapeutic schedule is good to maintenance hemodialysis patients with refractory secondary hyperparathyroidism.

Objective To investigate the best experimental scheme of Mycoplasma solid culture combined with liquid culture.Methods A total of 961 samples of urogenital tract excretion were collected from March 2016 to June 2016.Both solid culture and liquid culture were performed for detection of Mycoplasma,false positive broths were picked out after 48 h,20 μl of each one was transformed to solid media for subculture,final results were read after 48 h.The experimental data was analyzed to find an optimal combination culture scheme.Results The positive rate of solid culture was 38.7％ (372/961) for UU and 7.8％ (75/961) for MH.The positive rate of liquid culture was 45.27％ (435/961) for UU and 7.08％ (68/961) for MH.The different rate between both methods was 9.47％ (91/961) for UU (x2 =43.61,P＜0.005),and 3.02％ (29/ 961) for MH (x2 =1.24,P＞ 0.05).The different rate was 53.5 ％ (46/86) between primary solid culture and subculture.The positive rate of total (primary solid culture and subculture) solid culture was 41.9％ (403/961) for UU and 8.6％ (83/961) for M H,which produced higher sensitivity than single (primary solid culture or subculture) solid culture.The different rate between the total solid culture and the liquid method was 6.24 ％ (60/961) for UU (x2 =17.067,P＜0.05),and 3.43 ％ (33/961) for MH (x2=5.94,P＜0.05).Conclusion Perform both solid culture and liquid culture for Mycoplasma,pick out those false positive broths for subculture.Total solid culture could be more reliable as final result,for which could combine specificity of solid culture with sensitivity of liquid culture.

Objective To explore the influences of plumbagin on phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (Akt) signaling pathway in rats with carbon tetrachloride induced liver fibrosis.Methods Forty male SD rats were assigned to control group,model group,2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group,with 10 rats in each group.Rats of the control group were injected with 0.9％ NaCl solution (2 mL/kg) intraperitoneally,while rats of the other three groups were injected with 60％ carbon tetrachloride/peanut oil (2 mL/kg,3 times a week for 6 weeks) intraperitoneally.Since the third week after modeling,rats of plumbagin treated groups were treated with intraperitoneal injection of plumbagin at a dose of 2 mg/kg and 3 mg/kg (twice a week for 4 weeks),respectively.Serum levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),albumin (Alb) were monitored routinely.Hyaluronic acid (HA) and laminin (LN) were measured by radioimmunoassay after 6 weeks; protein expression of liver PI3K,Akt and phosphorylated Akt (p-Akt) were evaluated by Western blotting and immunohistochemistry,respectively.Comparison of means among groups was performed by univariate analysis of variance.Results The hepatic fibrosis model was successfully established after 6 weeks.Serum levels of ALT,AST,HA and LN of model group were significantly higher than those of control group (all P＜0.05).Serum levels of ALT,AST,HA and LN of 2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group were significantly lower than those of model group,and the differences were statistically significant (all P＜0.05).The protein expressions of PI3K and Akt in each group were comparable (all P＞0.05).The p-Akt protein was mainly expressed in nucleus of hepatocytes.The levels of p-Akt protein in control group,model group,2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group were 0.0821±0.0003,0.7374±0.0037,0.3679 ±0.0332 and 0.1327±0.0561,respectively,and that in model group was significantly higher than control group (t =851.302,P＜0.05),but those in plumbagin treated groups were both lower than model group (t=71.858 and 28.363,all P＜0.05).Conclusions Plumbagin presents anti-fibrotic effects in the liver,by down-regulating the expression of p-Akt during the development of fibrosis,which might be one of the antifibrotic mechanisms.

Objective To evaluate the therapeutic effects of recombinant expression plasmid containing hepatocyte growth factor (HGF) and augmenter of liver regeneration (ALR) on rats with hepatic fibrosis. Methods Ninety Sprague-Dawley rats, which had been established into hepatic fibrosis models, were equally divided into 6 groups: blank group, pcDNA3.1 therapy group,pcDNA3.1-HGF therapy group, pcDNA3. 1-ALR therapy group, pcDNA3.1-HGF and pcDNA3. 1-ALR combined therapy group, and pcDNA3. 1-HGF-ALR therapy group. Zero point one μmol of blank or plasmid was injected into model rats in each group by tail vein once a day for 3 days. Model rats in blank group didn't receive any treatment. Additional 10 rats were chosen as control group, which were not given any interference during the experiment. All rats were sacrificed 4 days after end of treatment. Liver tissues were reserved for observing pathologic changes after HE staining and detecting proliferating cell nuclear antigen (PCNA) and c-jun by immunohistochemistry. Measurement data were compared by single-factor analysis of variance. Comparison between groups was done by SNK test. Enumeration data were analyzed by Fisher's exact test. Results In blank group and pcDNA3.1 therapy group, hyperplasia of fibrous connective tissue was very obvious, false lobules were formed. There was no significant difference between these two groups (x2 =0. 317,P= 1. 000).In the 4 remaining groups, hepatic fibrosis all achieved different degree of amelioration, and the therapeutic effect of pcDNA3.1-HGF-ALR was optimal. In control group, the expressions of PCNA and c-jun in liver tissues were low, with absorbance value of 8.6±1.9 and 3.2 ± 1.2, respectively. In blank group and pcDNA3. 1 therapy group, the expressions of PCNA and c-jun were obviously increased, with absorbance value of 24. 1±3.0, 24.5±4.3 and 23.8±3.1, 24.9±4.2, respectively,which were significant different from control group (all P＜0.01). In the 4 remaining groups, the expressions of PCNA were all obviously increased, and expressions of c-jun were all obviously decreased. The maximum change scope was observed in pcDNA3. 1-HGF-ALR therapy group.Conclusions The recombinant expression plasmid pcDNA3. 1-HGF-ALR can effectively ameliorate experimental hepatic fibrosis of rats. The anti-fibrosis effects are achieved probably by up-regulating PCNA expression and down-regulating c-jun expression.

Objective To construct and identify an eukaryotic expression plasmid containing rat hepatocyte growth factor(rHGF)gene and rat augmenter of liver regeneration(rALR)gene,so that to provide experimental basis for developing new treatments of hepatic fibrosis.Methods The gene fragments of rHGF and rALR were amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR by polymerase chain reaction(PCR),respectively,then were spliced by overlap extension PCR with a linker,and the fusion gene rHGF-linker-rALR was constructed.The fusion gene was directionally inserted into the eukaryotic expression plasmid pcDNA3.1 between restriction sites of Kpn Ⅰ and Xba Ⅰ to construct the recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR,and the new constructed recombinant plasmid was identified by double restriction digestion and DNA sequencing.Results DNA fragments of 2200 bp and 400 bp were observed after the electrophoresis of products amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR,respectively,which was consistent with the theoretical value.The electrophoresis of fusion gene rHGF-linker-rALR obtained by overlap extension PCR technique showed only a 2 600 bp DNA fragment,which was in accordance with the expected value.Electrophoresis of products of pcDNA3.1-rHGF-linker-rALR digested with Kpn Ⅰ and Xba Ⅰ showed two DNA fragments with 2600 bp and 5400 bp,which were both consistent with the expected value.The sequences were confirmed correctly by DNA sequencing.Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR is successfully constructed,which provides experimental basis for developing gene therapy of hepatic fibrosis.

Objective To evaluate the effects of a single preoperative dose of gabapentin on buprenorphine patient controlled intravenous analgesia after modified radical mastectorny. Methods Sixty female patients,ASA physical starus Ⅰ and Ⅱ undergoing modified radical mastectomy under general anesthesia were divided into two groups of 30 each group by random digits table to receive either gabapentin 1200 mg (gabapentin group) or a matching placebo (control group), administered orally 2 h before the induction of anesthesia. Subjects received patient controlled intravenous buprenorphi(n)e analgesia during thepostoperative period. Fifty-three patients finished this study, 28 cases in control group,25 cases in gabapentin group. Postoperative pain (static and dynamic),postoperative nausea and vomiting,anxiety,sedation were assessed by pain visual analogue scale(VAS), four-point ordinal scale, anxiety visual analogue scale, Ramsay sedation scale respectively. Postoperative buprenorphine consumption and time to first patient controlled analgesia were observed. Results Postoperative VAS (static and dynamic) was lower in gabapentin group than that in control group (P ＜0.05). Postoperative buprenorphine consumption was (506.1 ±37.9)μg , time to first patient controlled analgesia was (21.1 ±2.3)min,incidence rate of postoperative nausea and vomiting was 40.0% (10/25), antemetic rate was 12.0% (3/25), grade of anxiety was (28.5 ± 12.1) scores in gabapentin group, (699.8 ± 87.8)μ g, (4.3 ±0.8) min,64.3% (18/28),32.1%(9/28) and (66.3±15.7) scores in control group respectively. There were significant differences between two groups (P ＜ 0.05). Conclusion A single preoperative oral dose of gabapentin 1200 mg can effectively attenuate postoperative pain,reduce the consumption of buprenorphine,decrease the incidence rate of postoperative nausea and vomiting,improve patients' anxiety in patients undergoing modified radical mastectomy under general anesthesia.

Objective To discuss the value of leukocyte-targeted myocardial contrast echocardiography (MCE) as a tool in observing the degree of acute rejection after heart transplantation. Methods Abdominal heterotopic cardiac transplantation was performed on 32 rats successfully, among which 8 isografts served as group A, and groups B, C and D involved 8 allografts respectively. The rats in groups B and C were treated with cyclosporine A (CsA) at a high dose (10mg· kg-1 · day-1 ), a low dose (3 mg · kg-1 · day-1 ) from 3rd day before transplantation respectively.The rats in groups A and D were untreated with CsA. MCE was performed during continuous intravenous SonoVue injection postoperatively on the third day after operation. We performed 2 types of MCE: perfusion imaging and leukocyte-targeted imaging. The images were obtained at 20 s and 5 min after injection of contrast agent. The value of the contrast image grayscale (GS) was measured by image analyzer (GS20s, GS5 min). GStarget was calculated as the GS5min minus the GS20s in the same rat.Postmortem histology was performed after observation. The degree of myocardial rejection was determined by HE-stained graft myocardium. Immunohistochemistry was performed to quantify the CD3-positive cells, and correlation analysis was performed between CD3-positive cell count and GS20s,GS5min, GStarget. Results Perfusion imaging showed no significant difference in myocardial GS20s of each group. Leukocyte-Targeted imaging exhibited a clear gradient in these groups (P＜0. 05). There was significant difference in GStarget of each group (P＜0. 001). Postmortem histology showed 0- Ⅰ grade rejection in group A, Ⅰ -Ⅱ grade rejection in group B, Ⅱ-Ⅲ grade rejection in group C, Ⅲ-Ⅳ grade rejection in group D. Immunohistochemistry revealed the CD3-positive cell infiltration was increased in turn from the group A to the group D. There was a significantly positive correlation between the CD3-positive cell count and GStarget ( r = 0. 86, P ＜ 0. 001 ). Conclusion Leukocyte-targeted contrast echocardiography may thus offer a noninvasive and effective ultrasound imaging technique for detecting the degree of acute cardiac transplant rejection.

Objective To explore the feasibility and potential application value of establishing the neonatal pig models of islet transplantation under the renal capsule. Methods Nine wild-type neonatal Duroc pigs were selected, including 1 animal as the control (p6307), 6 as islet transplant donors and 2 as islet transplant recipients (p6210, p6207). After islet isolation and differentiation in vitro, islet transplantation under the renal capsule of the pig was performed. Immunosuppressive therapy of tacrolimus (Tac) combined with sirolimus was given after operation. Postoperative body weight, blood glucose and serum creatinine levels of the recipients were monitored. The p6210 recipient neonatal pig was sacrificed at postoperative 4 weeks, while the p6207 recipient and the control neonatal pig were sacrificed at postoperative 8 weeks. The islet grafts under the renal capsule were collected for pathological staining and insulin immunofluorescent staining. Results After islet transplantation under the renal capsule of the pigs, the growth rate of body weight of the recipients was significantly slower than that of the control neonatal pig, accompanied with intermittent symptoms, such as anorexia and diarrhea, etc. However, the blood glucose and serum creatinine levels of the recipients did not significantly differ from preoperative levels and those of the control neonatal pig. Evident islet mass was observed under the renal capsule of the p6210 recipient. Pathological staining and insulin immunofluorescent staining confirmed that the islet mass had the function of secreting insulin, whereas no obvious islet mass could be seen under the renal capsule of the p6207 recipient. Pathological staining detected no evident islet mass, suggesting the possibility of islet transplantation failure caused by rejection in the p6207 recipient. Conclusions The establishment of neonatal pig models of islet transplantation under the renal capsule is a feasible technique, which provides preliminary evidence for the establishment of composite islet-kidney donor graft in pig models for xenotransplantation in the treatment of end-stage diabetic nephropathy.