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Objective To observe the clinical efficacy of Yuganlong capsule combined with entecavir in treatment of liver fibrosis in patients with chronic hepatitis B cirrhosis. Methods A total of 57 cases of patients with chronic hepatitis B cirrhosis were randomly divided into treatment group(30 cases given entecavir) and control group(27 cases treated with Yuganlong capsule combined with entecavir).Fibroscan values,hepatic fibrosis indexes(HA,LN,C-Ⅳ,PC-Ⅲ),liver function index(ALT,AST,GGT,T-BiL),and ultrasonic imaging index(inner diameters of portal vein,thickness of spleen) were detected. Results Before treatment, all indicatorshad no significant differences between the two groups(P>0.05).After 96 weeks of treatment,the Fibroscan value,HA, LN,C-Ⅳ,ALT,AST and portal vein diameter were significantly reduced in the treatment group as compared with those before treatment(all P<0.05),and there were significant differences between the treatment group and the control group after the treatment (all P<0.05). Conclusion The effect of Yuganlong capsule combined with entecavir in the treatment of chronic hepatitis B liver cirrhosis is better than that of using entecavir alone to improve liver function and liver fibrosis.
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Objective To investigate the dynamic expressions of interleukin-22(IL-22),Interleukin-22 receptor 1(IL-22R1),and hepatic stellate cells(HSC)senescence in mice with Schistosoma japonicum infection. Methods A murine model of S. japonicum infection was established and the serum samples and liver tissues were collected 4,6,8,12 weeks post-infection. The serum samples were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST). The pathological changes and proliferation of hepatic collagen fibers in the liver tissue were observed after HE staining and Masson staining. The HSC senescence in fibrotic livers was determined by the detection of senescence-associatedβ-galactosidase(SA-β-Gal). Sandwich ELISA was used to measure the expressions of IL-22,and Real-time PCR was used to test the mRNA levels of IL-22 and IL-22R1. The control group without S. japonicum infection was set up. Results The serum levels of ALT and AST signifi-cantly increased 8 weeks and 12 weeks after the infection(vs. 0 week,all P<0.05). The level of IL-22 increased 4 weeks and 6 weeks after the infection(vs. 0 week,both P<0.05),but reduced 8 weeks post-infection,and was even lower 12 weeks post-in-fection(vs. 4 weeks and 6 weeks,both P<0.01). Being consistent with the dynamic expression of IL-22 protein,the mRNA ex-pression of IL-22 began to increase 4 weeks and reached the peak 6 weeks after the infection(vs. 0 week,both P<0.05),and continuously declined 8 weeks and 12 weeks post-infections(vs. 6 weeks,both P<0.05). The increase of the expression of IL-22R1 mRNA was correlated with the progression of fibrosis,and the peak was in 12 weeks post-infections(vs. 0 week and 6 weeks,both P<0.05). The number of senescence-associated beta-galactosidase-positive HSCs was reduced with the decreasing expression of IL-22 in the advanced liver fibrosis. Conclusion IL-22 and IL-22R1 are involved in the pathogenesis of schistoso-miasis liver fibrosis. As an inflammation factor,IL-22 significantly increases in the early stage of fibrosis. The expression of IL-22 decreases in the late stage of fibrosis,which may contribute to HSC senescence and restrict liver fibrosis.
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Objective To investigate the anti-fibrogenesis property and mechanism of short hairpin RNA (shRNA ) targeting heat shock protein 47 (HSP47) in schistosomiasis liver fibrosis mice model . Methods Sixty female BALB/c mice (SPF level) were infected percutaneously with (16 ± 1) Schistosoma japonicum cercariae . Twelve mice which were not infected with Schistosoma japonicum were set as uninfected control .The 60 mice were randomly divided into five groups including 1 mg/kg HSP47-shRNA intervention group ,2 mg/kg HSP47-shRNA intervention group ,4 mg/kg HSP47-shRNA intervention group ,unrelated shRNA intervention group and infected control group ,with 12 mice each group .From 6 to 14 weeks post infection ,mice in HSP47-shRNA intervention groups were intravenously injected with HSP47-shRNA weekly and those in unrelated shRNA intervention group were intravenously injected with 2 mg/kg of unrelated shRNA vector weekly .Survival rate ,liver and spleen in general and liver histology of mice in each group were observed at week 14 which was considered as end of intervention . The expressions of HSP47 mRNA and protein were determined by real-time polymerase chain reaction (PCR) and Western blot .Collagen type Ⅰ ,collagen type Ⅲ ,tissue inhibitor of metalloproteinase-1 (TIMP-1) , matrix metalloproteinase-9 (MMP-9) ,plasminogen activator inhibitor-1 (PAI-1) ,transforming growth factor-β1 (TGF-β1 ) , connective tissue growth factor (CTGF ) , interleukin (IL )-13 and IL-17 at transcriptional level were measured by real-time PCR .Means among groups were compared using student-t test .Results In vivo intervention of HSP47-shRNA mainly localized in hepatic stellate cells of mice . The survival rate of 1 mg/kg ,2 mg/kg and 4 mg/kg HSP47-shRNA intervention groups were 25 .00% , 25 .00% and 33 .33% , respectively .Mice received 4 mg/kg dose of HSP47-shRNA had significantly higher survival rate than the infected controls (χ2 = 4 .168 ,P= 0 .043) .In both 4 mg/kg HSP47-shRNA intervention group and the infected control group , hematoxylin and eosin staining showed chronic granuloma change ,of which ova were wrapped by the spindle-shaped collagen and inflammatory cell infiltrated .The percentage of positive staining for Masson trichrome and sirius red showed the quantity of the collagen deposition was significantly reduced by the HSP 47-shRNA intervention ,compared to the infected control group (t = 3 .191 ,P = 0 .039) .HSP47-shRNA significantly reduced HSP47 expression , and reduced the transcriptional levels of collagen type Ⅰ ,collagen type Ⅲ ,TIMP-1 ,PAI-1 ,TGF-β1 , CTGF ,IL-13 and IL-17 (all P < 0 .01) ,and increased the expression of MMP-9 mRNA ( P < 0 .01) . Conclusion Delivery of shRNA targeting HSP47 to the hepatic schistosomiasis mice model can silence the expression of HSP47 and improve the liver fibrosis .Collagen degradation and related cytokines release through upregulation of MMP-9 and downregulation of PAI-1 may be one of the anti-fibrotic mechanisms in HSP47-shRNA intervention .
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The role of interleukin-17 (IL-17) in autoimmune hepatitis (AIH) was investigated. A mouse model of experimental autoimmune hepatitis was established, and the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally into adult male C57BL/6 mice. The IL-17 expression in serum and the livers of the mice models was detected by using ELISA and immunohistochemistry, respectively. IL-17 neutralizing antibody was used to study the biological effect of IL-17 in the experimental AIH. IL-17 neutralizing antibody in vivo administration alleviated the hepatic inflammation and ALT level in the AIH model. IL-17 in the peripheral blood mononuclear cells (PBMC) of AIH patients was measured by using real-time PCR method. The results showed that IL-17 level was significantly up-regulated in AIH patients and mice models. It was concluded that IL-17 contributed to the development of AIH and might be a potential therapeutic target of AIH.
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Objective To evaluate efficacy and mechanism of Anluohuaxian pilule combined with interferon-γ in the treatment of schistosomal liver fibrosis. To preliminarily study on the relationship of pigment deposition in liver and schistosomal liver fibrosis. Methods Thirty Kunming mice were divided into the normal control group, the infection control group and the combination of Anluohuaxian pilule and Interferon-γ treated group. Schistosomal liver fibrosis model was established by infection with 40 Schistosoma japonicum cercariae. The treated group was treated by combination of Anluohuaxian pilule and Interferon-γ for 8 weeks. The changes of pigment deposition and hepatic egg granuloma in Schistosoma japonicum infected mice were observed. Expressions of collagen Ⅰ and Ⅲ were detected by immunohistochemistry. Transforming growth factor (TGF)-β1 was detected by fluorescent polymerase chain reaetion(PCR). Histopathology and computer image analysis were applied to evaluate the change in the liver tissues. Results The amount of pigment deposition in liver was related to the expression of TGF-β1 mRNA (correlation coefficient = 0. 8). Compared to the infection control group, combination of Anluohuaxian pilule and Interferon-γ can lessen hepatic fibrosis(P<0.05). The combination therapy can also make pigment deposition less and hepatic granuloma smaller than the infection control group(P<0. 05). Conclusions Pigment deposition in liver is related to the expression of TGF-β 1. Combination of Anluohuaxian pilule and Interferon-γ can lessen hepatic fibrosis in mice infected with Schistosoma japonicum. It's one mechanism to of the combination therapy down-regulate the expression of collagen Ⅰ, Ⅲ and TGF-β 1.
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Differential gene expression profiles in Balb/cJ mouse model of acute hepatic failure infected with MHV-3 virus intervened by anti-hepatic failure compound (AHFC) and the changes of cytokines regulated by genes were investigated. The Balb/cj mice were divided into AHFC-intervened group and control group randomly. Acute hepatic failure model of Balb/cJ mice infected with MHV-3 virus was established. The survival rate in the two groups was observed. It was found that the survival rate in the AHFC-intervened group and control group was 90% and 50% respectively 48 h after intraperitoneal injection of MHV-3 (P<0.05). Before and after the experiment, the cytokines in peripheral blood of the survival mice were determined, and RNA was extracted from survival mouse liver tissue for the analysis of the differential gene expression by a 36 kb mouse oligonucleotide DNA array. In all the genes of microarray there were 332 genes expressed differently in the two groups, in which 234 genes were up-regulated and 78 genes down-regulated. Through clustering analysis, the differential expression of immune related genes, including TNF receptor superfamily, Kctd9, Bcl-2, Fgl2, IL-8, IL-6, IFN-gamma, TNF-alpha etc. might be related with the curative effectiveness of AHFC. It was suggested that AHFC can balance the immune state of mouse model of acute hepatic failure infected with MHV-3 virus mainly through regulating the expression of immune related genes, decrease the immune damage and inhibit liver cell apoptosis of mouse acute hepatic failure model obviously so as to increase the survival rate of mouse models of acute hepatic failure.
Subject(s)
Coronavirus Infections/complications , Drugs, Chinese Herbal/therapeutic use , Gene Expression Profiling , Hepatitis, Viral, Animal/complications , Liver Failure, Acute/drug therapy , Liver Failure, Acute/etiology , Liver Failure, Acute/genetics , Mice, Inbred BALB C , Murine hepatitis virus , Phytotherapy , Random AllocationABSTRACT
Differential gene expression profiles in Balb/cJ mouse model of acute hepatic failure in- fected with MHV-3 virus intervened by anti-hepatic failure compound (AHFC) and the changes of cytokines regulated by genes were investigated. The Balb/cj mice were divided into AHFC-intervened group and control group randomly. Acute hepatic failure model of Balb/cJ mice infected with MHV-3 virus was established. The survival rate in the two groups was observed. It was found that the survival rate in the AHFC-intervened group and control group was 90% and 50% re- spectively 48 h after intrapefitoneal injection of MHV-3 (P<0.05). Before and after the experiment, the cytokines in peripheral blood of the survival mice were determined, and RNA was extracted from survival mouse liver tissue for the analysis of the differential gene expression by a 36 kb mouse oli- gonuleotide DNA array. In all the genes of microarray there were 332 genes expressed differently in the two groups, in which 234 genes were up-regulated and 78 genes down-regulated. Through clustering analysis, the differential expression of immune related genes, including TNF receptor superfamily, Kctd9, Bcl-2, Fg12, IL-8, IL-6, IFN-γ, TNF-α etc. might be related with the curative effectiveness of AHFC. It was suggested that AHFC can balance the immune state of mouse model of acute hepatic failure infected with MHV-3 virus mainly through regulating the expression of immune related genes, decrease the immune damage and inhibit liver cell apoptosis of mouse acute hepatic failure model obviously so as to increase the survival rate of mouse models of acute hepatic failure.
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OBJECTIVE To investigate the changes in etiological factors and the diagnostic methods of fever of unknown origin(FUO).METHODS The clinical data were retrospectively studied from patients with FUO hospitalized in Tongji Hospital from Jan 1992 to Dec 2006.During this time,528 cases fulfilled the criteria of FUO.RESULTS Out of the 528 FUO cases,definite diagnosis was eventually achieved in 476 patients(90.2%).The most common causes of FUO were infectious diseases(54.9%),with tuberculosis accounting for 39.0%(113/290) of cases of infection.Seventy patients(13.3%) were suffered from malignancy;61 patients(11.6%) were suffered from connective tissue disease(CTD);and 55 patients(10.4%) were suffered from other diseases.However,no definite diagnosis had been made in the remaining 52(9.8%) cases until they discharged from the hospital.Pathological diagnosis(30.5%) was the most important diagnostic method,in addition constituent ratio of pathogenic microorganism detection,molecular biology identification technique,immunologic test,diagnostic therapy and imaging accounting for 21.6%,12.8%,12.2%,11.6% and 11.3%,respectively.CONCLUSIONS The causes and diagnostic method of FUO are altered in the past 15 years.Infectious diseases are still the most common cause of FUO,from which tuberculosis is the top one.The cases of fungal infection and AIDS are increased slowly.Malignancy is decreased and occupies the third place of FUO after rheu-matic diseases.Drug induced fever,various kinds of clinical symptoms and syndromes and multifactorial fevers are increased.Diagnostic methods and rate of definite diagnosis are improved.
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Objective: To investigate the relationship of murine fibrinogen-like protein 2(mfgl2) / fibroleukin with pulmonary impairment in the murine model of severe acute respiratory syndrome(SARS).Methods: The Balb/cJ mice infected with murine hepatitis virus strain 3(MHV-3) through the trachea were observed for the pathological features and virus distribution in different organs.The expressions of both mfgl2 and fibrino in the lungs were determined by in situ hybridization and immunohistochemistry.Results: The MHV-3 infected mice developed typical interstitial pneumonia with extensive hyaline membrane formation in the alveoli,presented micro-vascular thrombosis in the pulmonary vessels and died within 5 days.MHV-3 virus replication was identified in all the organs observed.The specific expression of mfgl2 prothrombinase was noted in the terminal and respiratory bronchioles,alveolar epithelia and infiltrating cells.Conclusion: The characteristics of the pulmonary impairment of SARS in human can be properly simulated by the MHV-3 induced murine model of SARS.The up-regulation of the specific gene mfgl2 in the lungs involved in fibrino deposition and microvascular thrombosis may be largely responsible for SARS-associated pulmonary damages.