Objective:To investigate the relationship between NOD-like receptor protein 3 (NLRP3) gene promoter region-30 single nucleotide polymorphism (SNP) (rs3738448) G/T and dilated cardiomyopathy (DCM) and its related risk factors.Methods:A case-control study method was used to collect 137 patients and 140 healthy controls; polymerase chain reaction-restriction endonuclease fragment length polymorphism technology combined with sequence alignment after DNA sequencing was used for data statistics; After Hardy-Weinberg balance test, the χ 2 test was used for correlation analysis; logistic regression was used to analyze the correlation between multiple risk factors and the SNP site and the incidence of DCM; SNPinfo database was used to predict and analyze the transcription factors affected by the SNP. Results:A total of GG and GT genotypes were detected at this SNP locus, and their genotype distributions were in line with Hardy-Weinberg equilibrium ( P>0.05). At the same time, the difference between the DCM group and the control group was significant ( P<0.05). Multivariate logistic regression analysis indicated that mean arterial pressure, C-reactive protein and B-type brain natriuretic peptide were independent risk factors for the onset of DCM (all P<0.05). The incidence of DCM in -30(RS3738448)G/T genotype GT group was 2.243 times higher than that in GG group (95% CI: 1.043-4.827, P<0.05). Logistic regression analysis under dominant, recessive and additive genetic models showed that there was a correlation between the dominant inheritance of SNP and the occurrence of DCM ( OR=0.44, AIC=370.4, BIC=381.3, P<0.05). Conclusions:The -30 (rs3738448) G/T SNP in the promoter region of the NLRP3 gene is associated with the pathogenesis of DCM, and provides population genetic data for the study of polymorphisms in the promoter region of NLRP3 gene.

ObjectiveTo investigate the clinical efficacy of 96-week entecavir (ETV) treatment for HBeAg-negative chronic hepatitis B patients with compensated cirrhosis, as well as its effect on patients with different HBV DNA loads. MethodsA total of 118 chronic hepatitis B patients with compensated cirrhosis who visited Xiangyang Hospital of Traditional Chinese Medicine from January 2009 to June 2013 were enrolled and all had Child-Pugh class A cirrhosis. According to HBV DNA load, these patients were divided into high-load group (group A, HBV DNA≥105 copies/ml) and low-load group (group B, HBV DNA＜105 copies/ml). All the patients were treated with ETV 0.5 mg/d for 96 weeks. Child-Pugh score was used to evaluate liver function before and after treatment, and the changes in alanine aminotransferase (ALT), albumin (Alb), and total bilirubin (TBil) after treatment were observed. Hyaluronic acid (HA), α2-macroglobulin, and liver stiffness measurement (LSM) were used to evaluate liver fibrosis. The t-test was used for comparison between groups, and a repeated-measures analysis of variance was used for comparison within one group and between groups at different time points. Chisquare test was applied for comparison of categorical data between the two groups. ResultsThe two groups showed significant reductions in ALT, Alb, and TBil at weeks 12 and 24 of treatment (all P＜0.05), and the ALT normalization rate and HBV DNA clearance rate at weeks 24 and 48 of treatment showed significantly differences between the two groups (χ2=9241，6428，11134，5139，all P＜0.05). Both groups showed significant reductions in HA, α2-macroglobulin, and LSM after treatment (t=2648，1921，4018，3166，2136，3461，all P＜0.05). The incidence rates of complications such as variceal bleeding, ascites, and hepatocellular carcinoma showed no significant differences between the two groups (all P＞0.05). ConclusionIn HBeAg-negative chronic hepatitis B patients with compensated cirrhosis, ETV can significantly improve liver function, slow down the process of liver fibrosis and cirrhosis, and effectively reduce long-term complications.

Objective To characterize the baseline status of Chinese diabetic patients based on data derived from Chinese cohort from SOLVETM study.Methods Patients with type 2 diabetes initiating basal insulin detemir at the decision of the physician were eligible for the study.Data on demographics,medical history,glycemic profile and treatment regimen at baseline were collected by physicians.Results A total of 3272 patients [female 42％,male 58％,mean age (56.2 ± 10.8) years] were included in the study.Their BMI was (25.3 ± 3.3) kg/m2.The duration of diabetes was 4.0 (0.1-27.0) years,and the duration of treatment with oral antidiabetic drugs (OADs) was 3.0(0.0-20.2) years.The proportions of subjects with diabetic macro-and micro-vascular complications were 15.8％ (515 cases) and 27.1％ (866 cases),respectively.The hemoglobin Al c (HbAl c) at baseline was (8.33 ± 1.70) ％,and the fasting blood glucose (FPG) was (9.5 ± 2.6) mmol/L.Conclusions A large proportion of patients with type 2 diabetes remain in poor glycemic control,and the prevalence of diabetic complications is high,which requires optimal therapeutic strategy for the patients with suboptimal glycemic control.

BACKGROUND: Dendritic cells play an important role in antigen present in vivo, and the mechanism of tumor cells in escaping the antigen presentation of dendritic cells existed in the patients with tumor.OBJECTIVE: To sensitize dendritic cells from human peripheral blood with apoptotic hepatoma cells induced by mitomycin.DESIGN: A randomized control trial by taking apoptotic hepatoma cell sensitized dendritic cells as the observed subjects.SETTINGS: Institute of Field Surgery, Daping Hospital, the Third Military Medical University of Chinese PLA; Institute of Radiation Medicine,Chinese PLA Academy of Military Medical Sciences.MATERIALS: The experiments were carried out in the Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences from April 1998 to May 1999. The cell strain was the QBC939 bile duct cancer cell strain, and mitomycin was used as the antitumor drug.METHODS: Mononuclear cells were isolated from the peripheral blood of normal people, 50μg/L granulocyte-macrophage colony stimulating factor(GM-CSF) and 1 000 U/mL interleukin-4 (IL-4) were added, once every other day for 4 times. On the 3rd day of culture, the apoptotic bile duct cancer cells induced by mitomycin was added, and then cultured in vitro for 4 days, finally the dendritic cells were collected.MAIN OUTCOME MEASURES: ① The identification of the cultured dendritic cells was observed; ② The dendritic cells were co-cultured with necrotic and normally cultured bile duct cancer cells respectively, and the phagocytized apoptotic body loaded antigens were observed; ③ The immunostimulatory activity of dendritic cells (1×103, 5×103 and 1×104/well)and that after loaded by antigen were detected, and the mononuclear cells were taken as controls.RESULTS: ① The cultured and amplified dendritic cells expressed high levels of costimulatory molecules of CD1a and B7, and there were typical irregular processes on the surface. ② The tumor cells formed apoptotic bodies when they were induced by mitomycin, which were arrested and phagocytized by dendritic cells. ③ The ability of the antigen loaded dendritic cells in stimulating the proliferation of allogenic lymphocyte T was further enhanced.CONCLUSION: The apoptotic tumor cells induced by mitomycin can induce the mononuclear cells from human peripheral blood differentiating into the dendritic cells with the concomitance of GM-CSF and recombinant IL-4 and amplify dendritic cells. Meanwhile, the dendritic cells can effectively present the antigens of apoptotic bile duct cancer cells, and it probably becomes a new effective approach for tumor antigen to sensitize dendritic cells.

OBJECTIVETo explore the survival time, pathological change and liver regeneration in different kinds of reduced-size liver transplantation in rats using steatotic grafts.

METHODSMacrovesicular and microvesicular steatotic rat liver models were established by feeding rats with a diet consisting of 79% standard chow, 20% lard and 1% cholesterol for different time periods. With modified two cuff vascular anastomoses and end-to-end sutures on the bile duct, reduced-size orthotopic rat liver transplantations were performed in an attempt to explore the ratio of graft weight to recipient body weight, recipient original liver weight and histological and electron-microscopic findings in comparison with whole rat liver transplantations.

RESULTSA one-week survival rates for the rats undergoing whole liver transplantation, and those in the 70% reduced-size orthotopic liver transplantation (ROLT) group, the 60%ROLT group and the 50%ROLT group (grade I macrosteatotic grafts) were 91.67%, 75%, 75% and 25%. A 2-week survival rate was 83.33%, 75%, 58.33% and 0 respectively. And their graft recipient body weight (GRBW) values SD were 3.56% +/- 0.36%, 2.53% +/- 0.15%, 2.28% +/- 0.12% and 1.83% +/- 0.16%, respectively. In grade II macrosteatotic grafts, the one-week survival rate for those undergoing whole liver transplantation and those in the 70% ROLT group was 83.33% and 25%. In the microsteatosis grafts for whole liver transplantation, 70% ROLT, 60% ROLT and 50% ROLT, the one-week survival rate was 83.33%, 75%, 75% and 33.33%; and the 2-week survival rate was 75%, 66.67%, 66.67% and 0, respectively. The survival rate of the 50% ROLT group using grade I macrosteatotic grafts or grafts mainly with microsteatosis was significantly different from that of other groups. While using macrosteatotic grafts in grade II, the 1-week survival rate of the 70% ROLT group was very poor. Pathological findings after operation included liver regeneration and portal space with mild lymphocyte infiltration. Improvement in steatosis and dilation of the central vein and sinusoids was observed in some rats.

CONCLUSIONSIn the successful and long-term survival of rat reduced-size liver transplantation using grade Imacrosteatotic grafts or grafts with microsteatosis, the GRBW values should be over 2.28% +/- 0.12%, and the value of graft-recipient liver weight should be over 60%. Steatotic livers in grade II should not be used as grafts in ROLT. Steatosis was improved and even totally cured in some long-term survival rats.

Animals ; Fatty Liver ; complications ; Graft Survival ; Liver Transplantation ; methods ; Male ; Rats ; Rats, Wistar

Objective To explore the effect of fatty liver on graft survival, especially with reference to macrovesicular and microvesicular steatosis and to evaluate the relationship between histological grading and inflammation activity. Methods Different degrees of rat fatty liver model were established by feeding rats a diet consisting of 79% standard diet, 20% lard and 1% cholesterol. By modified two cuff vascular anastomoses and end to end suture for bile duct, rat orthotopic liver transplantation was performed to evaluate the relationship between donor histological grading and survival rate. Results Low survival rate of macrosteatosis (grade Ⅲ) was found. Most rats died of liver failure in early days after transplantation. Pathological findings showed frequent hepatic necrosis. There was no significant difference between macrosteatosia(gradeⅠ) and the normal group. After transplantation, almost all of the fat was cleared by the end of the fourth week. Diminished steatosis and liver regeneration were found in macrosteatosis (gradeⅡ), while microsteatotic donors had higher survival rate than the other groups except the normal group. Conclusion Macrovesicular steatosis(grade Ⅲ) affects graft survival and these steatotic livers should not be used as donors. However, steatotic livers with mild macrovesicular steatosis (grade Ⅰ) and microvesicular steatosis(grade Ⅲ) do not influence recipient survival, so these livers can be used safely for liver transplantation. The ischemic damage should be considered when using livers of macrovesicular steatosis(gradeⅡ). Donors with numbering score more than 2.7 are correlated with the poor survival.

Objective To determine the effect of diffusion chamber for immune isolation and the proliferation of allogenic hybridoma within diffusion chamber in mice. Methods The hybridoma cell strain was established from SP2/0 cells and the splenocytes of BALB/c mouse by PEG mediate technique. The cells were injected peritoneally into BALB/c ( n =5) and C57 BL/6 (B6, n =5) mice. Diffusion chambers with microporous membrane containing hybridoma cells were implanted peritoneally into 5 BALB/c and 5 B6 mice. Results Within two months after peritoneal injection of the cell strain, four BALB/c mice developed bloody ascites and one developed abdominal tumor, but the B6 mice developed neither ascites nor tumor. After 30 and 101 days of the chamber implantation, the chambers were coated by omentum or mesenterium in all the animals of both strain groups and freshly formed blood vessels to the chamber membrane could be observed. Within the chambers, tumor developed, but the tumor could not fully fill the inner space of the chamber during the whole observation period. Conclusion The hybridoma cells established from SP2/0 cells and the splenocytes of BALB/c mouse may carry BALB/c MHC antigen. The diffusion chambers can effectively isolate immune rejection. Freshly formed blood vessels to the chamber membrane can support the cells within the chambers.

Objective To investigate the effects of antisense TIMP 1 on the interstitial collagen synthesis of fibrotic liver in rats. Methods Fibrosis was induced in the liver of rats with the injection of 60% CCl 4 and 5% alcohol. Recombinant antisense TIMP 1 gene mammalian expression vectors were construct and infused into the fibrotic liver of rats. The expressions of TIMP 1 mRNA, ?1(Ⅲ) mRNA and protein were determined by RT PCR, ISH and immunohistochemistry. The serum levels of PCⅠ, PCⅢ in rats were determined by RIA. Results Antisense TIMP 1 gene could markedly suppress the expression of endogenous TIMP 1 mRNA in the transfected liver of rats( P

Objective To observe the survival time, pathological change and liver regeneration in different kinds of reduced size liver transplantation in steatotic rats. Methods Rat models of different kinds of reduced size orthotopic liver transplantation were performed by modified two cuff vascular anastomoses and end to end suture for bile duct to observe the recipient body weight, graft weight, recipient original liver weight, histological and pathological and electron microscopic findings in comparison with those of the whole rat liver transplantation. Results One week survival rate of the whole liver transplantation, 70% reduced size liver transplantation(ROLT), 60% ROLT and 50% ROLT group (grade Ⅰsteatotic donor) was 91.67%, 75%, 75% and 25%, respectively, and 2 week survival rate was 83.33%, 75%, 58.33% and 0, respectively. In grade Ⅱ steatotic donor, 1 week survival rate of the whole liver transplantation and 70% ROLT was 83.33% and 25%. As to donor livers with microvesicular steatosis, 1 week survival rate of the whole liver, 70% ROLT, 60%ROLT and 50% ROLT was 83.33%, 75%, 75% and 33.33% and the 2 week survival rate was 75%, 66.67% 66.67% and 0. The survival rates of 50% ROLT in grade Ⅰ steatotic donor and livers mainly with microvesicular steatosis were significantly different from those in other groups. The 1 week survival rate of 70% ROLT was very poor in steatotic donors in grade Ⅱ. Pathological findings after operation included liver regeneration and mild lymphocyte infiltration in portal space, the amelioration of the steatosis in some cases and dilation of the central vein and sinusoids. Conclusion To obtain long term survival of reduced size liver transplantation using steatotic donors, the GRBW should be over (2.28?0.12)(the ratio of graft to recipient liver weight over 60%). Steatotic livers in grade Ⅱ should not be used as donors in ROLT. The steatosis can be ameliorated after operation.

OBJECTIVETo investigate the effect of ursodeoxycholic acid (UDCA) on liver regeneration after 70% partial hepatectomy (PH) in bile duct obstructive (BDO) rats.

METHODSWistar rats were randomly divided into N-PH group in which normal rats were operated with 70% PH, BDO-PH group in which 70% PH were operated after two week's BDO, and BDO-PH UDCA or sterile saline treatment group in which UDCA (15mg kg(-1) d(-1)) or saline was administrated during BDO and after 70% PH. The hepatic pathological changes were observed. BrdU labeling of hepatocytes, the mRNA expression of intrahepatic hepatocyte growth factor (HGF) and its receptor (Met gene) after 70% PH were measured by immunohistochemical analysis and RT-PCR, respectively.

RESULTSImprovements of hepatic function and pathological changes were induced by UDCA administration after BDO. The expression of hepatic HGF/Met mRNA after 70% PH in BDO-PH UDCA treatment group rats was significantly increased compared with N-PH group rats (P<0.05), BrdU peak labelling of hepatocytes (59.39% +/- 10.82%) in BDO-PH UDCA treatment group rats was significantly higher than that (36.22% +/- 8.37%) in BDO-PH group rats (t=4.149, P<0.01) and without significance compared with N-PH group rats (68.64% +/- 11.26%, t=1.451, P >0.05).

CONCLUSIONSUDCA promotes liver regeneration after 70% PH in BDO rats by remission of hepatic pathological changes and elevating hepatic mRNA expression of HGF and Met.

Animals ; Cholestasis ; genetics ; physiopathology ; surgery ; Gene Expression Regulation ; drug effects ; Hepatectomy ; Hepatocyte Growth Factor ; genetics ; Liver ; physiology ; surgery ; Liver Regeneration ; drug effects ; Male ; Proto-Oncogene Proteins c-met ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Wistar ; Ursodeoxycholic Acid ; pharmacology