ABSTRACT
Acute liver failure (ALF) is a syndrome characterized by rapid onset, rapid progression, and poor prognosis. Although much progress has been achieved in the etiology and pathogenesis of ALF and emergency liver transplantation and comprehensive treatment have significantly increased the treatment success rate, there are still many difficult issues in clinical practice. This article reviews the research advances in the classification, etiology, main clinical types, prognostic scoring system, and treatment of ALF in recent years.
ABSTRACT
Objective To observe the nuclear translocation of transcription factor NF-κB and IRF-3 in TLR4 silenced EVC304 cells infected by HTNV and to provide new information for anti-HTNV innate immunity and its signal transduction. Methods TLR4~- cells and TLR4~+ cells were infected by HTNV 76-118, respectively. The cells stimulated by LPS were selected as positive control groups, and the cells without stimulation were selected as negative control groups. After 6 hours, indirect immunofluorescence assay(IFA) was used to detect the nuclear translocation of NF-κB and IRF-3. Results The transcription factor NF-κB and IRF-3 transfered into nuclear 6 hours after stimulated by HTNV 76-118. Conclusion TLR4 may mediate the nuclear translocation of transcription factor NF-κB and IRF-3 in HTNV infected human umbilical vein endothelial cells.
ABSTRACT
Objective To develop a reverse genetics system for Hantaan virus (HTNV) 84FLi strain by using RNA polymerase [ (pol Ⅰ)-mediated transcription. Methods Complementary DNA (cDNA) containing the coding sequence for chloramphenicol acetyhransferase (CAT) was inserted into the 5'-and 3'-terminal untranslated regions of HTNV 84FLi L segment. These chimeric cDNAs (pol Ⅰ expression cassette) were cloned into plasmids and between the human pol Ⅰ promoter and terminator to generate sense and anti-sense RNA pol Ⅰ transcription reporter plasmids. The reporter plasmids were transfeeted into 293T cells or the 1:1 combination of 293T and HTNV infected Vero cells. These cells were cotransfected with expression plasmids encoding Ⅰ. (RNA dependent RNA polymerase) and N (nucleoprotein) viral proteins, Cells were harvested 48 h post-transfection and the CAT activity was detected. The 293T cells were infected with the supernatant to explore the passage ability of CAT activity. ResultsThe reporter plasmids pLvRNA-CAT and pLcRNA-CAT were constructed successfully. CAT activity was detected in transfected cells and could also be serially passaged in the rescued virus minigenomes. Conclusion The RNA polymerase ]-driven reverse genetics system successfully rescues HTNV 84FLi minigenomes.
ABSTRACT
Objective To detect circulating CD4 + CD25 + regulatory T cells (Treg) and Toll-like receptor(TLR)2 and TLR4 expression on the peripheral blood mononuclear cells (PBMCs) of patients with HBV-related liver cirrhosis (LC), and to explore the correlation between them. Methods PBMCs isolated from 30 LC patients, 21 chronic hepatitis B (CHB) patients and 16 normal controls(NC) were stained with fluorescent labeling anti-TLR2-PE, anti-TLR4-APC, anti-CD14-FITC monoclonal antibodies and anti-CD4-PerCP, anti-CD25-FITC, anti-CD127-PE. Samples were detected by flow cytometry. Statistic analysis be-tween groups was performed by Kruskal-Wallis H test. Spearman rank correlation was used to analyze the correlation of Treg and TLR2, TLR4. Results The expression of TLR2 and TLR4 were significantly up-reg-ulated in patients with LC than those in the controls (TLR2 : 200.3 ± 96.8 vs 94.1 ± 17.6, P < 0.05 ; TLR4:32.1 ±7.2 vs 17.8 ±3.9, P<0.05). The expression of TLR4 was significantly increased in pa-tients with LC than those in patients with CHB (TLR4 : 32. 1 ± 7.2 vs 25.2 ± 8.3, P < 0.05), but there were no differences of TLR2 expression between LC and CHB(200.3 ± 96.8 vs 214.0 ± 72.6, P > 0.05). Treg/CD4+ T cells were 5.07% ±1.43%, 5.88% ±1.66%, 4.21% ±1.24% in patients with LC, CHB and NC, respectively. Treg/CD4+ T cells were significantly increased in patients with CHB than those in pa-tients with NC(P<0. 05) and LC(P <0.05), but there were no differences between LC and NC(P > 0.05). TLR4 expression and Treg were positive correlation (r = 0. 469, P = 0. 032) and TLB2 expression were negative correlation in patients with LC (r = -0.428, P = 0.021). Conclusion The expression of TLR2 and TLR4 were up-regulated on PBMCs in patients with LC. It seems to be expression of TLR2 and TLR4 in-volved in the pathogenesis of LC.
ABSTRACT
Objective To observe the protective effects of Toll-like receptor(TLR)-4 siRNA against acute liver injury in mice induced by lipopolysaccharide(LPS)and D-galactosamine(D-GalN).Methods One hundred and fifty C57BL/6 male mice were divided into 5 groups: phosphate buffered solution(PBS)pretreatment group,negative control plasmid pretreatment group,TS4 pretreatment group,TS6 pretreatment group and TS7 pretreatment group.Acute liver injury was induced in mice by intraperitoneal coinjection of LPS(10 ng/g)and D-GalN(1 mg/g).In vivo delivery of siRNA was performed via the tail vein by hydrodynamic injections(50 μg siRNA dissolved in 1 mL PBS)24 h and 48 h before coinjection of LPS and D-GalN. Expression of TLR-4 in liver tissues was measured by immunohistochemistry.The changes of TLR-4,tumor necrosis factor(TNF)-α and macrophage nflammatory protein(MIP)-2 mRNA levels in liver tissues were determined by reverse transcriptasepolymerase chain reaction(RT-PCR)analysis.MIP-2 and TNF-α concentrations in the sera of mice were determined by enzyme-linked immunosorbent assay(ELISA). Levels of alanine transaminase (ALT) and aspartate transaminase(AST) in serum were measured by standard autoanalyzer techniques. Liver pathological changes were observed by haematoxylin-eosin staining, while cell apoptosis levels in liver were determined by terminal deoxynucleotidyl-mediated-dUTP nick end labeling (TUNEL)assay. The difference of survival rates in 5 groups was analyzed by Fisher's exact probability test.ResultsPretreatment with TLR-4 siRNA down-regulated the TLR-4 mRNA and protein expressions,and significantly decreased the mortality and liver injury caused by coinjection of LPS and D-GalN in C57BL/6 mice.TLR-4 siRNA significantly down-regulated the TNF-α and MIP-2 mRNA expression and cytokine levels as determined by RT-PCR and ELISA,respectively. TLR-4 siRNA abrogated hepatocyte necrosis and inflammatory infiltration and also remarkably reduced serum concentrations of transaminases. The percentage of TUNEL-positive hepatocytes was significantly reduced in TLR-4 siRNA pretreatment group(TS4 pretreatment group: 0.065±0.015 vs PBS pretreatment group; 0.346±0.062,P＜0.05).ConclusionIt suggest that inhibition of TLR-4 expression by TLR-4 siRNA may provide potential application value for preventing liver injury.
ABSTRACT
<p><b>BACKGROUND</b>To investigate the biological effects of anti VEGF₁₆₅ ribozyme on human lung adenocarcinoma cell.</p><p><b>METHODS</b>Hammerhead ribozyme (VRz) against VEGF₁₆₅ gene transcripts (site 212) and its paired mutant ribozyme (mVRz) were designed and synthesized, and the cleavage activity of the ribozymes on target RNA in a cell-free system was observed. The replication-incompetent adenovirus-mediated eukaryotic expression vectors (rpAdVRz) containing VRz and mVRz gene were constructed and identified. Then the human lung adenocarcinoma cells (A549) were infected with recombinant adenovirus. The biological characteristics of A549 cell before and after infection in vitro were inspected by Northern blot, laser confocal imaging system analysis, flow cytometry and transmission electron microscopy.</p><p><b>RESULTS</b>VRz specifically and efficiently cleaved the VEGF₁₆₅ mRNA. The rpAdVRz was successfully constructed and infected A549 cell. The level of VEGF₁₆₅ expression decreased 87% in rpAdVRz infected cells compared with the other groups, but their biological characteristics were not influenced by the expression of the exogenous gene.</p><p><b>CONCLUSIONS</b>The adenovirus mediated hammerhead ribozyme against VEGF₁₆₅ can significantly decrease the expression of VEGF₁₆₅. This provides an experimental basis for human lung cancer gene therapy with antiangiogenesis method.</p>
ABSTRACT
Nucleoside analogues are currently the main drug therapy against HBV infection. However, this type of therapy can cause gene mutations in HBV that result in drug resistance. The types of mutations caused by nucleoside analogues and the molecular mechanism of in vitro drug-resistance were reveiwed.
ABSTRACT
Objective:To observe the therapeutic effects of plasma exchange in severe hepatitis. Methods:53 cases served as treatment group and 49 cases as control group. Both groups were similar in basic medical treatment, and an additional treatment of plasma exchange was carried in the treatment group. 186 times of plasmas exchange were performed in the treatment group, to observe and compare the changes in patients of the 2 groups in symptoms、liver functions and survival rates. Results:The clinical symptoms of patients in the treatment group were obviously improved, the liver functions were also obviously improved in the treatment group as compared with the control group, and the survival rate of treatment group was higher than that of control group(73.58% vs 46.94%, P
ABSTRACT
Current therapies for chronic hepatitis caused by hepatitis B virus (HBV) are limited. RNA interference (RNAi) may constitute a new therapeutic strategy for various hepatitis virus infections. In this review we present the advances in inhibition of HBV by RNAi, the unresolved questions and therapeutic potential of RNAi.
ABSTRACT
The study on bioartificial liver support system was widely accepted in recent years. It's mainly containing material-hepatocytes. This summary is concerning about the research and application of hepatocytes in this field.
ABSTRACT
Hantavirus is the main cause of hemorrhagic fever with renal syndrome (HFRS). It is an acute infectious diseases characterized by fever, hemorrhage, nephritis or thrombocytopenia, and hantavirus pulmonary syndrome(HPS). The main clinical manifestations are fever, hemorrhagic lesion, acute respiratory distress and capillary leakeage syndrome. These are four different serotypes of the hantavirus species: Hantan virus(HTNV),Seoul virus(SEOV),Dobrava/Belgrade virus(DOBV),and Puumala virus(PUUV). They are known to cause HFRS, while Sin Nombre virus(SNV) causes HPS. In China, these are two serotypes of hantavirus: HTNV and SEOV found. The severity of infection depends on the viral serotype. To find a safe, rapid and specific serotyping diagnosis of the causative virus is important. The results not only can be beneficial for rodent control, but also for prevention and therapy. The current research of Hantavirus nucleocapsid protein used as serotyping antigen are summarized.
ABSTRACT
<p><b>OBJECTIVE</b>To determine the frequency and characteristics of reassortment among Hantaan and Seoul viruses causing hemorrhagic fever with renal syndrome (HFRS).</p><p><b>METHODS</b>Mixed infections were initiated in tissue culture, using Hantaan virus strain 76 - 118 and Seoul virus strain SR-11. Potential reassortant virus plaques were picked out by multiplex RT-PCR, using primers specific for individual genome segments (L, M, S) of each strain.</p><p><b>RESULTS</b>Most of the progeny virus plaques (68.19% of 44) had parental genotype of 76 - 118 strain or SR-11 strain while 2 of 44 plaques had mixed genotypes that yielded RT-PCR bands for the same segment of both parental strains. Reassortant viruses were detected in 68.19% of 44 progeny plaques tested, involving the M and S segments. In addition, approximately 4.55% of the progeny virus plaques appeared to contain S or M segments originating from both parental virus strains, showing that they were diploid.</p><p><b>CONCLUSION</b>Genetic reassortment can occur between Hantaan virus and Seoul virus strains.</p>
Subject(s)
Animals , Chlorocebus aethiops , Genome, Viral , Genotype , Hantaan virus , Genetics , RNA, Viral , Genetics , Reassortant Viruses , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Seoul virus , Genetics , Vero CellsABSTRACT
Objective:To study the different effect of fused gene IL-2/Fc administrated at different time on immune responses induced by HBV preS_2S genetic vaccine.Methods:BALB/c mice were immunized with purified recombinant plasmids at 0, 2, 4 weeks. Two groups were designed. One group were injected with pcDNA3.1S_2S at first, then pcDNA3.1IL-2/Fc was administrated after three days. While the other group were injected two plasmids at the same time. We compared the immune responses through detecting the anti-HBs titers, CTL cytotoxicity level, proliferation of lymphocytes and cytokines levels secreted by cultural splenocytes.Results:The data demonstrated that the humoral and cellular immune responses induced by pcDNA3.1IL-2/Fc as adjuvant and administrated after HBV preS_2S DNA vaccine 3 days mice were dominant higher than other groups.Conclusion:Genetic adjuvant administrated after Ag priming can enhance its effect.
ABSTRACT
The coding region of S genome segment of Hantaan virus (76/118 strain) was inserted into the eukarytic expression plasmidpVR1012. The recombinant expression plasmid pVRS22 was constructed. Vero-E6 cells were transiently transfected in vitro with pVRS22 plasmid. The transient expression of Hantaan virus nucleocapsid proteins in Vero-E6 cells was detected by indirect immunofluorescence assay (IFA) with monoclonal antibody 5H5 against Hantaan virus.