Objective:To investigate the effect and mechanism of Fuzheng Jiedu Tongluofang on the expression of MMP-2,MMP-9 and invasion of human hepatoma HepG2 cell.Methods:Hepatioma HepG2 cell was treated with Fuzheng Jiedu Tongluofang.The cell viability was measured by CCK8.The invasion of HepG2 was detected by Transwell assay.The expression of MMP-2 and MMP-9 was analyzed by ELISA.Results:In the CCK8 assay,Fuzheng Jiedu Tongluofang could inhibit the proliferation of HepG2 cells at a density and time measure.In the Transwell assay,the inhibitory rate of invasion was 52.45% when treated with Fuzheng Jiedu Tongluofang at a concentration of 2.65 mg/ml. The ELISA assay indicated that the expression of MMP-2 and MMP-9 decreased significantly after treated with Fuzheng Jiedu Tongluofang(P<0.05). Conclusion:Fuzheng Jiedu Tongluofang suppressed the invasion of HepG2 with the possible mechanism of down-regulating the expression of MMP-2 and MMP-9 in HepG2.

AIM:To construct the adenovirus vector with adiponectin (Acrp30) siRNA, and to observe its effect on the Acrp30 expression and glucose transport in 3T3-L1 adipocytes. METHODS: Mouse Acrp30 siRNA fragment was designed, synthesized and cloned into the adenovirus vector. 3T3-L1 cells were infected with the two recombinant adenoviruses, respectively. The mRNA expression and protein levels of Acrp30 in these cells were evaluated by RT-PCR and ELISA. Glucose transport was measured by 2-Deoxy-[3H]-D-glucose incorporation method. RESULTS: The recombinant adenoviruses were successfully constructed. They remarkably downregulated the expression of Acrp30 at both mRNA and protein levels in 3T3-L1 cells, and decreased the glucose transport in 3T3-L1 adipocytes (P

Objective To investigate the clinicopathological characteristics of HBV recurrence after liver transplantation. Methods The retrospective analysis of the clinicopathological changes was performed on 17 patients who had HBV recurrence after liver transplantation in our medical department. Results HBV recurrence happened from 4 to 48 months. Twelve of them which were identified to be YMDD mutation switched to entecavir or added adefovir. Three of them receiving chemotherapy when liver cancer recurred switched to entecavir. Two of them with withdrawal of lamivudine were given lamivudine continuously. Liver function returned to the normal level and HBVDNA was ＜ 102 U/ml after anti-hepatitis B virus. The histological changes in the transplanted livers included hepatocellular degeneration, necrosis and apoptosis, portal infiltrations and fibrosis.With time after recurrence, it was easier to see hepatitis B virus replication in liver cells, incidence of acute rejection, increases of liver fibrosis and the formation of fibrous septa, even pseudolobule.Conclusion In native HBV infection livers, fibrosis occurs more early and develops rapidly. The number of virus is closely related to liver necrosis and inflammation. Early discovery and change to quick and effective treatment of anti-hepatitis B virus in time can improve greatly the prognosis of the patients.

Chitosan is a polymer with good biocompatibility, unique pH-responsive solubility, convenient modification and easier film-formability. Chitosan could serve as an active mediator between biological components and microfabricated devices to prepare biological micro electro mechanical systems (BioMEMS) with high selectivity and sensitivity. Recently, there has been a growing interest in BioMEMS based on biofabrication of chitosan. We reviewed the mechanisms and processes of three biofabrication methods based on chitosan, including directed assembly, enzymatic assembly and self-assembly. Current applications and research progress in biological, medical and environmental fields are also discussed. Finally, future research directions are prospected.
Biocompatible Materials ; chemistry ; Biomimetics ; Biosensing Techniques ; Catalysis ; Chitosan ; chemistry ; Micro-Electrical-Mechanical Systems ; methods ; Nanotechnology ; Nucleic Acid Hybridization

Objective:To further explore the relationship between Type Ⅰ hypersensitivity reaction and the mechanism of the viral hepatitis B through the research of soluble intercellular adhesion molecular-1(sICAM-1) and soluble vascular cell adhesion molecular-1(sVCAM-1) levels in serum in patients with viral hepatitis B related to Type Ⅰ hypersensitivity reaction and the damage of liver cells, and therefore, provide new theory for the perfection of the immunological mechanism of hepatitis B, especially acute hepatitis B.Methods:Serum sICAM-1 and sVCAM-1 levels were measured in 45 patients with viral hepatitis B and 15 normal cases using double antibody sandwich ELISA method. ALT and AST levels were studied using omni automatic biochemistry analyzer and its correlation with sICAM-1 and sVCAM-1 were observed.Results:The sICAM-1 and sVCAM-1 levels of 7 acute hepatitis B were significantly higher than that of the normal cases. ②The sICAM-1 and sVCAM-1 levels of 38 chronic hepatitis B were also significantly higher than that of the normal cases. ③The sICAM-1 and sVCAM-1 levels of 13 cases moderate chronic hepatitis B were significantly higher than that of 16 cases mild chronic hepatitis B. ④The sICAM-1 and sVCAM-1 levels of 9 serious chronic hepatitis B were significantly higher than that of the group of mild chronic hepatitis B. ⑤Levels of sICAM-1 were significantly positively correlated with serum ALT and AST. ⑥sVACM-1 levels were significantly positively correlated with serum ALT and AST. In conclusion, sICAM-1 and sVCAM-1 levels in patients with acute hepatitis B were most remarkably increased, and then serious chronic hepatitis B, moderate chronic hepatitis B, light chronic hepatitis B in sequence.Conclusion:Serum sICAM-1 and sVCAM-1 levels in patients with viral hepatitis B may reflect the damage of liver. ②Examination of serum sICAM-1 and sVCAM-1 levels in patients with hepatitis B can be used to judge the patient’s condition and to diagnose. ③As an important indication of the inflammatory in Type Ⅰ hypersensitivity reaction, sICAM-1 and sVCAM-1 may interfere the appearence of hepatitis B and the procedure of immunological damage of liver cells.

Objective:To present evidence for the pathogenetic role of allergic factor,histamine,in type I allergy for induction of liver damage.Methods:Three groups of rabbits were fed normally and injected (iv) daily with 0, 0.04 or 0.08 ?g/kg phosphohistamine, respectively, for days. The serum level of ALT and AST in each group rabbits was assayed dynamically during the treatment. After treatment for days, the tested rabbits were sacrificed for pathological examination of the liver tissues.Results:The serum level of both ALT and AST in rabbits treated with phosphohistamine increased significantly during the tested periods, compared to that of the control group. However, both ALT and AST levels showed no significant difference between 0.04 ?g/kg and 0.08 ?g/kg groups. Liver microscopic examination, pathological damage could be observed in the tested groups in a time-and dose-dependent manner under microscopic examination. No evident pathological change appeared in the control group.Conclusion:Liver damage could be induced by histamine dosage-and time-dependently. This pathological action of histamine, a type I allergic factor, presents further evidence for a direct role of type I allergy in the pathogenesis hepatic injury.

Objective:To construct a fusion protein of extracellular domain peptide fragment of muscle specific kinase ( MuSK) and fluorescent protein mCherry ,and used as antigen in the detection of antibodies against MuSK ( MuSKAb ) in the sera of patients with myasthenia gravis ( MG).Methods:The mCherry gene was amplified by PCR from vector pRSET-B and cloned into pGEM-T Easy Vector,and furthermore, cloned into Eukaryotic expression vector pMT /BiP/V5-His ( MuSK), which contains MuSK extracellular domain 22-452 amino acid peptide fragment gene to construct the fluorescent fusion protein gene MuSK -mCherry.The recombinant vector was subsequently transfected into drosophila S 2 cells for expression.The expressed fusion proteins were verified in confocal mi-croscope ,and used as antigen in the detection of MuSKAb in sera of MG patents in fluorescence immunoprecipitation test .Results:The fluorescent fusion protein MuSK-mCherry was successfully constructed and expressed.The MuSKAb in sera of patents with MG could be detected in fluorescence immunoprecipitation test using the constructed MuSK-mCherry fusion protein as antigen.Conclusion: It is available to use the constructed fluorescent fusion protein MuSK-mCherry as antigen in fluorescence immunoprecipitation test for the detection of MuSKAb in sera of patents with MG.

ObjectiveTo investigate the association between different hepatitis C virus (HCV) genotypes and serum interleukin-17 (IL-17), interleukin-6 (IL-6), and vitamin D in patients with HCV-related cirrhosis. MethodsSeventy-six patients with HCV-related cirrhosis, who were admitted to 302 Hospital of PLA from January to December, 2012, were enrolled in the study, and they were divided into type 1b group (n=47) and type 2a group (n=29) according to the genotypes. The levels of serum IL-17, IL-6, and vitamin D were determined by enzyme-linked immunosorbent assay. Comparison between the two groups was made by t test, and Spearman correlation analysis was used to investigate the association between serum IL-17, IL-6, and vitamin D and genotypes. ResultsThe findings in improved Child-Pugh classification showed that there were significant differences in the percentage of grade A patients between type 1b group and type 2a group (χ2=4.97, P＜0.05); when compared with type 2a group, type 1b group showed significantly higher lentiviral titers of alpha-fetoprotein (AFP) and HCV RNA and levels of IL-17, IL-6, and tumor necrosis factor-α (TNFα) (t=21.56, 16.51, 1231, 10.71, and 7.23, respectively, all P＜0.05), but significantly lower levels of interferon-γ (IFNγ), 25(OH)D, and 1,25(OH)2D (t=3.98, 6.32, and 4.88, respectively; all P＜0.05); Spearman correlation analysis showed that the lentiviral titers of AFP and HCV RNA and the levels of IL-17 and IL-6 were positively correlated with genotype 1b and genotype 2a (all P＜0.05), while the levels of IFNγ, TNFα, and 25-OH-D were negatively correlated with genotype 1b and genotype 2a (all P＜0.05). ConclusionCompared with those with genotype 2a, HCV-related cirrhosis patients with genotype 1b have higher serum levels of IL-17, IL-6, AFP, and HCV RNA, but a lower level of vitamin D; the results suggested that there are correlations between serum IL-17, IL-6, 25-OH-D, AFP, and HCV RNA and genotype, but no significant correlation between 1,25(OH)2D and genotype.

Objective:To investigate the association of single nucleotide polymorphisms (SNPs) of receptor-associated protein at the synapse ( rapsyn ) with myasthenia gravis ( MG ).Methods: The genomic DNA was extracted from peripheral blood cells , sampled from 132 patients with MG and 153 control individuals.The 8 exons of rapsyn gene were amplified by PCR ,then the products of PCR sequenced directly.Each sequence was compared with wild-type rapsyn gene , and the association between mutation and clinical symptoms of MG analysed.Results:No mutation was found in the exons 1,2,4,5,6,7,and 8 of rapsyn gene both in MG patients and control group compared with the wild-type rapsyn gene.However,a new SNP,L222R[CTG>CGG(2)] or T665G,was found in exon-3.The allele and genotype frequencies of SNP L 222R met Hardy-Weinberg genetic equilibrium (P>0.05),indicating the group repre-sentativeness.The allele frequencies of G were not statistically different between patient and control groups ( P>0.05 ).There were differences in the 3 genotypes TT , TG and GG between patient ( 42.4% vs 48.5% vs 9.1%) and control ( 49.0% vs 33.3% vs 17.6%) groups ( P<0.05 ).The genotype frequencies of GG were statistically higher in control group than that in patient group , showing a recessive model of inheritance.Conclusion: The SNPs in the rapsyn gene are associated with MG in this study.L222R ( T665 G) is a new SNP found and allele G might be a protective factor for MG.

OBJECTIVETo develop an oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer using attenuated Salmonella typhimurium and evaluate its efficacy and protective effect.

METHODSThe eukaryotic expression vector including the MG(7)-Ag mimotope and a Th epitope was constructed, and then transduced into an attenuated Salmonella typhimurium to get the oral DNA vaccine. C57BL/6 J mice were orally immunized with 1 x 10(8) cfu Salmonella transfectants, with Salmonella harboring empty plasmid, with phophate buffered saline (PBS) as control. At the 6th week, serum titer of MG(7) antibody was detected by ELISA. In the 8th week, a [(3)H]-thymidine incorporation assay was performed to test the proliferation of murine spleen cells to the stimulant of MG(7)-Ag mimicry peptide. At the same time, Ehrlich ascites carcinoma cells expressing MG(7)-Ag were used in tumor challenge assay to evaluate the protective effect of the immunization.

RESULTSThe oral DNA vaccine induced MG(7) antibody in mice, while in vivo unprimed proliferation assay of the spleenocytes showed no difference among the three groups. Two weeks after tumor challenge, 2 in 7 immunized mice were tumor free, while none in the control group was protected.

CONCLUSIONOral DNA vaccine based on the MG(7)-Ag momitope is immunogenic. It is able to induce specific immunity response against tumor in mice, and the vaccine is partially protective.

Administration, Oral ; Amino Acid Sequence ; Animals ; Antigens, Neoplasm ; blood ; genetics ; immunology ; Base Sequence ; Cancer Vaccines ; genetics ; immunology ; therapeutic use ; Epitopes ; genetics ; immunology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Mimicry ; genetics ; immunology ; Molecular Sequence Data ; Plasmids ; genetics ; Polymerase Chain Reaction ; Stomach Neoplasms ; drug therapy ; immunology ; Treatment Outcome ; Vaccines, DNA ; genetics ; immunology ; therapeutic use