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OBJECTIVE To study the effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone （GA） on doxorubicin （DOX）in the treatment of liver cancer. METHODS A tumor bearing model was established by inoculating liver cancer cell H 22 into the right axillary skin of ICR mice. The successfully modeled mice were randomly divided into model control group，DOX group （2.5 mg/kg，once every other day ，via tail vein ），GA group （30 mg/kg，once a day ，gavage）and GA+DOX group（the usage was the same as single drug groups ），with 6 mice in each group. The administration cycle was 15 days. The general growth of mice in each group were observed ，body weight and tumor weight were measured ，and the inhibition rate of tumor was calculated. Jin’s formula was used to evaluate the effect of combined medication （Q）. The serum level of alpha-fetal protein（AFP），the pathological changes of tumor tissue ，cell apoptosis and the expression of platelet-endothelial cell adhesion molecule-1（CD31）were detected in each group. The cardiac index，serum levels of B-type natriuretic peptide （BNP）and N-terminal pro-brain natriuretic peptide （NT-pro BNP ），pathological changes of heart and myocardial fibrosis degree were also detected. RESULTS The percentage of body weight change （except for GA group ） and tumor weights of DOX group，GA group and GA + DOX group were all decreased significantly，compared with model control group （P＜0.05 or P＜0.01），while tumor weight of GA+DOX gro up was significantly lower than DOX group （P＜0.01）. Inhibitory rates of tumor in 3 administration groups were 54.29％，42.50％ and 89.29％ respectively，and Q of two-drug combination was 1.21. The tumor tissues of mice in each administration group were necrotic to varying degrees ；the serum level of AFP and the expression of CD31 in tumor tissue were decreased significantly ，compared with model control group （P＜0.05 or P＜0.01）；the percentage of necrosis area of tumor tissue and the positive rate of apoptosis （except for single drug groups ）were significantly increased （P＜0.05 or P＜0.01），while positive rate of apoptosis in GA+DOX group was significantly higher than DOX group （P＜0.05）. Cardiac index of mice in DOX group was significantly lower than model control group （P＜0.01）；serum levels of BNP and NT-pro BNP in DOX group and GA+ DOX group were significantly higher than model control group （P＜0.05 or P＜0.01）；pathological changes of heart and the degree of myocardial fibrosis in GA+DOX group were lower than DOX group. CONCLUSIONS GA combined with DOX show synergistic antitumor effect. GA can strengthen the apoptosis promoting effect of DOX ，and can help to reduce the cardiotoxicity of DOX.
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OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.
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Animals , Humans , Mice , Carcinoma, Hepatocellular/pathology , Cyclophosphamide/therapeutic use , Hemolysin Proteins , Heterografts , Interleukin-2 , Liver Neoplasms/pathology , Panax notoginseng , Saponins/therapeutic use , Tumor Necrosis Factor-alphaABSTRACT
Objective:To investigate the protective effect of IL-22 on rat liver ischemia reperfusion injury (IRI) and the potential mechanisms.Methods:Eighteen male specific pathogen free SD rats (7-8 weeks, about 250g) were randomly divided into three groups: Sham group (Sham), hepatic ischemia reperfusion (IRI) and IL-22 preconditioning group (IL-22+ IRI), respectively. The liver IRI model of 70% rats was established. The IL-22+ IRI group was intraperitoneally injected with rcIL-22 (50 mg/kg) 1 hour before surgery, and the Sham group and IRI group were injected with the same dose of normal saline 1 hour before surgery. After 1 h ischemia and 6 h reperfusion, blood was collected from the abdominal aorta, then liver tissue, serum aspartate transaminase (AST) and alanine aminotransfease (ALT) levels were measured. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver tissue were detected. The expression of signal transducer and activator of transcription 3 (STAT3), p-STAT3, nuclear factor erythorid-2 related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) were detected by Western blot. Results:Compared with Sham group, serum AST [(1 923.50±92.63) U/L, (1 004.25±65.05) U/L)] and ALT [(1 172.51±180.31) U/L, (583.50±164.75) U/L] levels were increased in IRI group and IL-22+ IRI group (AST: F=293.62; ALT: F=30.33, P<0.05). The levels of MDA in IRI group and IL-22+ IRI group [(1.72±0.12) μmol/mg, (0.98±0.05) μmol/mg] in liver tissue were higher than those in Sham group (0.58±0.14) μmol/mg protein ( F=186.73, P<0.05), and the expression of p-STAT3, Nrf2 and HO-1 was increased. SOD level in IRI group (28.51±3.85) U/mg was lower than that in Sham group (70.25±5.64) U/mg protein ( F=203.41, P<0.05). Compared with IRI group, serum AST and ALT levels in IL-22+ IRI group were decreased, SOD activity in liver tissue was increased, MDA level was decreased, and p-STAT3, Nrf2 and HO-1 expression was increased (all P<0.05). Conclusion:IL-22 could alleviate liver IRI in rats, and the mechanism may be related to the activation of STAT3 and Nrf2/HO-1 signaling pathway and anti-oxidative stress.
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Liver fibrosis is the result of persistent inflammatory response and chronic scar healing response during chronic liver injury and may progress to liver cirrhosis, portal hypertension, and liver failure, which finally requires liver transplantation. Interleukin-22 (IL-22) belongs to the IL-10 family and is the only cytokine that is produced by immune cells but does not act on immune cells. IL-22 plays a role by binding to its receptors IL-22R1 and IL-10R2, which has attracted much attention in the field of liver disease research in recent years. IL-22 not only plays the role of anti-inflammation and promotion of liver regeneration and tissue repair, but also has a pro-inflammatory effect in liver diseases, and it exerts a protective effect on the liver by reducing fibrosis in some pathological conditions, but there are still controversies over its association with liver fibrosis. IL-22 has different effects and mechanisms in liver fibrosis caused by different etiologies. This article reviews the role and possible mechanisms of IL-22 in liver fibrosis caused by viral infection (HBV and HCV), alcohol, high-fat diet, and autoimmunity.
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Objective To investigate the regulation of miR-939-5p on USP22 gene expression and its effect on liver cancer migration and proliferation.Methods The expression of miR-939-5p in hepatoma cell lines (HepG2,MHCC-97H,SMMC-7721,BEL-7404 and Huh7) and normal liver cell line LO2 was detected by realtime PCR (qPCR).The liver cancer cells with the lowest expression were used as experimental subjects,and transfected with miR-939-5p (Experimental group) or miR-NC (Control group).qPCR was used to detect the transfection efficiency of miR-939-5p.Transwell migration assay and MTT proliferation assay were used to detect the migration and proliferation of hepatoma cells after transfected miR-939-5p.Bioinformatics software predicted target genes for miR-939-5p.The dual luciferase reporter gene verified the interaction of miR-939-5p with the target gene.qPCR and Western blotting were used to detect the expression of target genes at mRNA and protein levels after over-expression of miR-939-5p.Measurement data were expressed as (Mean ± SD),and t test was used for comparison between groups.Results The expression of miR-939-5p was significantly lower in hepatoma cell lines than in normal hepatocytes (P ＜0.01),and the expression of miR-939-5p was the lowest in SMMC-7721 cells (P＜0.01).miR-939-5p was efficiently transfected into SMMC-7721 cells [(1.01 ±0.07) vs (20.12 ± 1.27),P ＜0.01].High expression of miR-939-5p inhibited the migration ability (P ＜ 0.01) and proliferative capacity of liver cancer SMMC-7721 cells (P ＜0.05).The USP22 gene may be a target gene of miR-939-5p.The luciferase reporter gene confirmed that miR-939-5p specifically binds to the 3'-UTR of USP22 mRNA (P ＜ 0.01).USP22 expression was decreased at mRNA and protein levels after high expression of miR-939-5p (P ＜ 0.01).Conclusions The expression of miR-939-5p was down-regulated in hepatocellular carcinoma cell lines,and miR-939-5p inhibited the migration and proliferation of liver cancer SMMC-7721 cells.The molecular mechanism was to interfere with the expression of USP22 gene.
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Objective To observe the sensitization of lidocaine on subcutaneous hepatoma H22-bearing mice and abdominal cavity H22 tumor-bearing mice treated by mitomycin. Methods According to the random number table method, the mice were divided into subcutaneous tumor-bearing group and abdominal cavity tumor-bearing group, with 15 mice in each group. The mice in the two groups were further divided into three subgroups: model group, mitomycin group, mitomycin+lidocaine group, with 5 mice in each subgroup. The day before the intraperitoneal injection, the density of H22 cells obtained from peritoneal culture of one mouse was adjusted to 5 ×106/ml. Subcutaneous tumor-bearing group mice were injected H22 cells into the right armpit, and abdominal cavity tumor-bearing group mice were injected H22 cells into the abdominal cavity, 0.2 ml per mouse. Intraperitoneal injection was given after inoculation for 24 h (the experiment day 1), followed by intraperitoneal injection on day 5 and 9. Univariate ANOVA analysis and t test were used to analyze the solid tumor weight and tumor inhibition rate on the 11th day of subcutaneous tumor-bearing mice, and the survival time and life extension rate within 60 days of abdominal cavity tumor-bearing mice. Results The solid tumor weight of subcutaneous tumor-bearing mice model group, mitomycin group and mitomycin + lidocaine group were (3.77 ±1.02) g, (1.67 ±0.28) g, (0.74 ±0.19) g, respectively, and the differences in the three groups were statistically different (F = 31.753, P < 0.01); compared with the subcutaneous model group, the subcutaneous solid tumor weights of mitomycin group and mitomycin +lidocaine group were decreased and the differences were both statistically different (t=2.10, P<0.01; t=3.04, P<0.01); the subcutaneous solid tumor weight of mitomycin+lidocaine group was lower than that of mitomycin group (t= 0.93, P= 0.034). The tumor inhibition rate of mitomycin group and mitomycin +lidocaine group reached 55.70％ and 80.37％ respectively. The survival time of abdominal cavity tumor-bearing mice in model group, mitomycin group and mitomycin + lidocaine group was (16.80±0.84) d, (28.80± 6.30) d, (40.40±12.86) d, respectively, and the differences in the three groups were statistically different (F=10.155, P=0.003); compared with the abdominal cavity tumor-bearing mice model group, the survival time of mice in mitomycin group and mitomycin + lidocaine group was prolonged (t= 12.00, P= 0.041; t= 23.60, P= 0.001), and it was found that survival time in mitomycin + lidocaine group was longer than that in mitomycin group (t=11.60, P=0.047). The life extension rate of mitomycin group and mitomycin+lidocaine group reached 71.43％ and 140.48％ respectively. Conclusion Lidocaine can increase the sensitization of mitomycin on hepatoma H22-bearing mice.
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Objective To investigate the association between interleukin-22 (IL-22) single nucleotide polymorphisms (SNPs) and the prognosis of hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF).Methods The patients with HBV-ACLF from the First Affiliated Hospital of Fujian Medical University were retrospectively studied.Seven SNP genotypes of IL-22 gene,including rs2227478,rs2227491,rs1179251,rs1179249,rs2227473,rs2227484,and rs11611206,were detected using imLDRTM multiple SNP typing kit and the distribution features of SNP genotypes were described.The relationship between the distribution of SNP genotypes and alleles and the prognosis of ACLF was analyzed.Comparison of genotypes and allele frequencies between groups were performed by chi-square test of R × C table or Fisher's exact tests.Binary logistic regression analysis was used to analyze whether IL-22 gene polymorphisms was an independent prognostic factor for patients with ACLF.Results A total of 122 patients with HBV-ACLF were included in this study.Ninety-two (75.1％) were male and 30 (24.59 ％) were female.Patients were stratified as survival group (90 cases) and non-survival group (32cases) according to the Results of three months follow-up.The genotype distribution of rs2227484 of IL-22 gene was significantly different between the two groups (x2=6.128,P=0.033).The A allele frequency in the non-survival group (15.6％) was significantly higher than that in the survival group (5.6％) with statistically significance (OR=0.318,95％ CI=0.126-0.804,P=0.012).There was no significant difference in the other six SNP genotypes of IL-22 gene between the two groups (all P＞0.05).However,binary logistic regression showed that rs2227484 of IL-22 gene was not an independent risk factor for the short-term mortality in HBV-ACLF patients (adjusted OR=3.102,95％ CI:0.939-10.250,P=0.063).Conclusions The A allele and AA genotype of rs2227484 of IL-22 gene may be associated with a short-term prognosis in patients with HBV-ACLF.
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BACKGROUND: Exposure to sustained high concentrations of HCFC-123 is known to be hepatotoxic. We report two simultaneous cases of toxic hepatitis related to exposure to 2,2-dichloro-1,1,1-trifluoroethane (HCFC-123), a common refrigerant, at a Korean fire extinguisher manufacturing facility. CASE PRESENTATION: Patients A and B were men aged 21 and 22 years, respectively, with no notable medical histories. They had recently started working for a manufacturer of fire extinguishers. During the third week of their employment, they visited the emergency center of a general hospital due to fever, lack of appetite, and general weakness. At the time of their visit, they were suspected as having hepatitis due to elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), and total bilirubin levels and were hospitalized. However, as their condition did not improve, they were moved to a tertiary general hospital. After conservative treatment, one patient improved but the other died from acute hepatic failure. Assessments of the work environment showed that the short-term exposure levels of HCFC-123 for valve assembly processes were as high as 193.4 ppm. A transjugular liver biopsy was performed in patient A; the results indicated drug/toxin-induced liver injury (DILI). Given the lack of a medical history and the occupational exposure to high levels of HCFC-123, a hepatotoxic agent, the toxic hepatitis of the workers was likely related to HCFC-123 exposure. CONCLUSIONS: Work environment assessments have not included this agent. To the best of our knowledge, we are the first to report a case of death related to HCFC-123-induced liver damage. Our findings suggest that exposure standards and limits for HCFC-123 must be developed in Korea; work environments will have to be improved based on such standards.
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Humans , Male , Alanine Transaminase , Alkaline Phosphatase , Appetite , Aspartate Aminotransferases , Bilirubin , Biopsy , Chemical and Drug Induced Liver Injury , Emergencies , Employment , Fever , Fires , Hepatitis , Hospitals, General , Korea , Liver , Liver Failure, Acute , Occupational Exposure , TransferasesABSTRACT
Objective@#To investigate the clinical features of imbalance between Th1 and Th22 cells and its association with disease progression in patients with liver cirrhosis, and to explore immune therapeutic strategies for targeted therapy for liver cirrhosis.@*Methods@#In vitro peripheral blood mononucleated cells (PBMCs) were collected by centrifugation. CD3-BV500 and CD8-PerCP-Cy5.5 staining was performed for these cells. IFNγ-PE-Cy7, IL-17a-APC, IL-22-PE, or the corresponding isotype control was added, and then PBMCs were fixed with 1% polyoxymethylene after being washed once by permeabilization-wash buffer. Flowjo software was used for the analysis of T lymphocyte subsets and cytokines. Th1 (CD4+IFNγ+), Th17 (CD4+IL-17a+), Th22 (CD4+IL-22+), Tc1 (CD8+IFNγ+), Tc17 (CD8+IL-17a+), and Tc22 (CD8+IL-22+) subsets were defined and the secretions of interferon-γ (IFN-γ), interleukin-17a (IL-17a), and interleukin-22 (IL-22) were measured for all subsets. LX-2 cells were cultured in a serum-free medium and different concentrations of recombinant human IL-22 protein (25, 50, 100 ng/ml) were added; 24 hours later, the activation marker α-smooth muscle actin (α-SMA) was used to measure LX-2 activation. Fetal bovine serum with a volume fraction of 10% was used as a positive control. Enzyme-linked immunosorbent assay (chemiluminescence) was used to measure the concentrations of hyaluronic acid, type III precollagen, and type IV collagen in supernatant. A one-way analysis of variance, the non-parametric Mann-Whitney U test, and the non-parametric Kruskal-wallis H test were used for statistical analysis based on data type.@*Results@#Compared with the health control group, the liver cirrhosis groups with various causes had significant increases in peripheral Tc1, Th17, and Th22 cells. The percentage of Th17 cells in the liver cirrhosis group was 1.64 times that in the control group (4.25%±2.45% vs 2.59%±1.36%, P < 0.05), and the mean percentage of Th22 cells in the liver cirrhosis group was 2.18 times that in the control group (4.17%±2.55% vs 1.31%±0.64%, P < 0.05). The percentages of Th17 (5.89%±3.44%) and Th22 cells (5.32%±3.67%) in the patients with alcoholic cirrhosis were 1.27 and 3.06 times those in the control group (P < 0.05). The patients with alcoholic cirrhosis had a significant increase in Th22 cells. The patients with different types of liver cirrhosis had a significant reduction in the ratio between anti-fibrotic and pro-fibrotic factors (Th1/Th22), which was positively correlated with the severity of liver cirrhosis and was a common immunological feature of liver cirrhosis with different causes. In addition, IL-22 activated hepatic stellate cells and promoted the production of collagen.@*Conclusion@#The imbalance between anti-fibrotic and pro-fibrotic factors (Th1/Th22) is a common feature of the progression of liver fibrosis with various causes and may contribute to the progression of liver fibrosis.
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Objective@#To investigate the effect of interleukin-22 (IL-22) on the activation and proliferation of hepatic stellate cells (HSCs) induced by acetaldehyde, as well as the role of the antioxidant axis Nrf2-keap1-ARE.@*Methods@#Hepatic stellate cell-T6 (HSC-T6) cells were cultured in vitro, and after 24 and 48 hours of acetaldehyde stimulation at various concentrations (25, 50, 100, 200, and 400 μmol/L), MTT assay was used to measure cell proliferation rate to screen out the optimal conditions for model establishment. HSC-T6 cells were treated first with the optimal concentration of acetaldehyde (200 μmol/L) for 24 hours and then with different concentrations of IL-22 (10, 20, and 50 ng/ml) for 24 hours. MTT assay was used to measure cell proliferation, Western blot and cell immunohistochemistry were used to measure the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and α-smooth muscle actin (α-SMA), and spectrophotometry was used to measure the changes in the content of malondialdehyde (MDA) and reduced glutathione (GSH) in culture supernatant. SPSS 17.0 was used for statistical analysis and data were expressed as mean±SD. P < 0.05 was considered statistically significant. A one-way analysis of variance was used for comparison of means between any two groups.@*Results@#HSCs had significantly enhanced proliferation and activation after being treated with acetaldehyde, especially at 200 μmol/L for 48 hours. After the intervention with gradient concentrations of IL-22, the proliferation and activation of HSCs were inhibited in a dose-dependent manner, and the proliferation and migration rates in the 10, 20, and 50 ng/ml IL-22 groups were 14%, 25%, and 35%, respectively (all P < 0.05). The results of Western blot and immunohistochemistry showed that there was no significant difference in the expression of Nrf2 total protein in HSCs between groups, while there was extremely low expression of Nrf2 nucleoprotein in the blank control group. There was increased expression of Nrf2 nucleoprotein after acetaldehyde stimulation (compared with the blank control group, P < 0.05), and after the intervention with gradient concentrations of IL-22, the expression of Nrf2 nucleoprotein was further increased (all P < 0.05). The results of spectrophotometry showed that compared with the blank control group, the model group had increased levels of MDA and GSH in culture supernatant after acetaldehyde stimulation; after the intervention with gradient concentrations of IL-22, there was a significant reduction in the MDA level and a significant increase in the GSH level in a dose-dependent manner (all P < 0.05).@*Conclusion@#The activation and proliferation of HSCs induced by acetaldehyde helps with the successful establishment of an in vitro model of alcoholic liver fibrosis. IL-22 effectively inhibits the activation and proliferation of HSCs induced by acetaldehyde, and its mechanism may be related to promoting Nrf2 nuclear translocation in HSCs and expression of the downstream target gene GSH and increasing the activity of the antioxidant axis Nrf2-keap1-ARE.
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Objective To study the promoting effects and mechanisms of interleukin-22 on liver regeneration in GCl4-induced liver fibrosis mice after partial hepatectomy.Methods One hundred and fortyfour C57/BL6 mice were randomly divided into four groups:PHX group,CCl4 group,CCl4 + PHX group,and CCl4 + IL22 + PHX group.The blood samples were taken to measure serum ALT and AST levels.ALT /AST was calculated to observe the liver injury at 3 h,6 h,12 h,24 h,48 h and 72 h after hepatectomy.The liver tissue specimens were collected at each time point after hepatectomy.We measured the hepatic lobe to calculate the liver weight ratio and conducted pathological examinations to observe the degree of fibrosis and pathological changes at each time point.The positive expression of proliferating cell nuclear antigen (PCNA) in liver tissue was tested by immunohistochemistry.The level of CyclinD1 and STAT3 (Signal transducer and activator of transcription 3) signaling pathway was detected by Western blot.Results (1) Compared with CCl4 + PHX group,the ALT/AST ratio of CCl4 + IL22 + PHX group was significantly higher at 24 h,48 h and 72 h,and the level of ALB of CCl4 + IL22 + PHX group was obviously increased at 48 h and 72 h (P ＜ 0.05).(2) The liver regeneration was significantly increased in CCl4 + IL22 + PHX group.Compared with CCl4 + PHX group (2.08 ± 0.16,2.77 ± 0.07,2.97 ± 0.14),the liver weight ratio of CCl4 + IL22 + PHX group(2.34 ± 0.07,3.23 ± 0.09,3.55 ± 0.09) dramatically increased at 24 h,48 h and 72 h.Moreover,the pathological sections displayed that the disease was alleviated (P ＜ 0.05).(3) Immunohistochemical assay and western blot revealed that compared with other three groups,the level of PCNA,STAT3 and Cyclin D1 was significantly lower in the CCl4 + PHX group.However,the level of PCNA,STAT3 and Cyclin D1 apparently increased in CCl4 + IL22 + PHX group at 24 h,48 h and 72 h (P ＜ 0.05).Conclusion Interleukin-22 may significantly promote liver regeneration and reduce liver pathological injury in liver fibrosis mice induced by administration of CCl4 after hepatectomy,which plays a positive role in the recovery of liver function.
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Objective To investigate whether a novel long-acting tumor necrotic factor (TNF) antagonist (soluble TNF receptor:IgG Fc [sTNFR:IgG-Fc]) can protect hepatocyte damage against liver failure caused by drugs in immunity-induced cirrhotic rats.Methods Wistar rats were repeatedly sensitized by human serum albumin (HSA) emulsified in complete freud adjuvant.The blood was collected at day 10 after the final sensitization.If anti-albumin antibody was positive,the rats were intravenously injected with HSA twice a week.After six weeks,liver cirrhosis was induced by immunity.All the model rats were divided into three groups with 15 each.Liver failure was induced with D-galactosamine/ lipopolysaccharide (LPS) intraperitoneal injection in the rats with liver cirrhosis in model group.The rats in pretreatment group were intraperitoneally injected with long-acting soluble TNF receptor p55 18 h before D-galactosamine/LPS injection.The control group were injected with 0.9％ sodium chloride.General condition,survival rate,liver function and pathological changes were all examined.Serum levels of interleukin (IL)-6,IL-22 and intrahepatic level of IL-6 were detected by enzyme linked immunosorbent assay (ELISA).The activity of Caspase 3 in hepatocyte lysis solution was measured by spectrophotography.Real-time polymerase chain reaction (PCR) was used to detect mRNA expressions of proliferating cell nuclear antigen (PCNA),bcl-2,bax and IL-22 receptor.Data were analyzed by variance analysis among groups.Results Rats in model group were dispirited with poor response after 12 hours and only 3 survived,compared with soluble TNF receptor p55 pre-treated group rats,in which all survived (P=0.029 8) with flexible response.Serum alanine aminotransferase levels in these two groups were (6 533± 360) and (105 ± 7) U/L,respectively.Hepatic regenerative nodule developed massive or submassive necrosis with septal fibrosis in model group,whereas soluble TNF receptor p55 alleviated the inflammatory and necrosis reaction of hepatic tissue.Serum IL-6 levels in model group and pretreatment group were (842.0±12.9) and (91.9±1.6) pg/mL,respectively (F=380.30,P＜0.01).Intrahepatic levels of IL-6 in these two groups were (26.2±1.2) and (11.1±0.8) pg/mL,respectively (F=176.90,P＜0.01),and serum IL-22 levels were (167.0±27.8) and (988.0±109.6) pg/mL,respectively (F=37.91,P＜0.01).Hepatic Caspase-3 activity was reduced by almost 60％ by soluble TNF receptor p55 pretreatment (F=303.70,P＜0.01) and bax expression reduced by 22％ (F=108.80,P＜0.01),while bcl-2 and PCNA expressions were up-regulated by 3.6-folds and 23.0-folds,respectively (F=115.60,P＜0.01; F=594.20,P＜0.01).Conclusions Long acting soluble TNF receptor p55 could improve survival rate,liver function and reduce inflammatory reaction of rats with liver failure induced by drugs on the basis of liver cirrhosis caused by immunity,which indicates that this drug may process a potential therapeutic value.
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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 explore the physical/chemical properties and effect of photodynamic therapy (PDT) of iminodiacetic acid modified tetraphenylporphyrin on mice bearing H22 liver cancer.Methods The UV absorbance spectrum,singlet oxygen yield and lipid-water partition coefficient were measured by UV spectrophotometry.Mice bearing H22 liver cancer as animal model were divided into control group (no drug,no light),light treated group (no drug,only light),drug treated group (only drug,no light),experimental once group (with drug and light,once) and experimental twice group (with drug and light,twice,PDT twice).Tumor inhibition rate and index of liver,spleen,lung,thymus of the mice were calculated to evaluate the antitumor activity of iminodiacetic acid modified tetraphenyl porphyrin-PDT.Results The iminodiacetic acid modified tetraphenylporphyrin had absorption at 650 nm,while its singlet oxygen yield and lipid-water partition coefficient were 2.30 and 0.52,respectively.No antitumor effect on mice bearing H22liver cancer for light treated group and drug treated group respectively.However,significant antitumor effect (P＜0.001)was found at experimental group.The maximum tumor inhibition rate could be 95.15％ for PDT twice group.No significant differences were observed on weight gain and liver,lung,spleen,thymus index in each group.Conclusion Iminodiacetic acid modified tetraphenylporphyrin with high singlet oxygen yield,low toxicity and significant in vivo anti-tumor activity,is suitable to be developed as an anti-tumor drug candidate for photodynamic therapy.
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OBJECTIVE: To explore the anti-tumor activities of a new compound 3β,12β,20(S)-trihydroxy dammarane-3-O-β-D-glucopyranosyl (1-2)-β-D-glucopyranoside (HRG), which is a substance of ginsenoside structure modification, in vivo. METHODS: Liver cancer H22 tumor model on the chick chorioallantoic membrane (CAM) was established to observe the effects of HRG on tumor growth, the number of induced vessels and the growth inhibition rate. The implanted tumors were dyed by hematoxylin-eosin (HE), and the morphological properties were studied with light microscope. Immunohistochemistry (SP method) was used to detect the expressions of the implanted tumor's MVD and VEGF. RESULTS: HRG inhibited the tumor growth at 25, 50 and 100 μg · mL-1. Compared with the model control group, the inhibitory rates of the tumor growth were 27.73%, 50.02%, and 64.21%, respectively. HRG significantly reduced the number of tumor-induced vessels. At the same time, there existed different degree necrosis among the treatment groups, and the degree of necrosis had an inverse relationship with the number of tumor-induced vessels. In addition, HRG reduced the MVD of the implanted tumors, and decreased the expression of VEGF. CONCLUSION: HRG has good anti-tumor activities in vivo, and can significantly inhibit the growth of liver cancer H22-CAM tumor and the induction of angiogenesis. The mechanisms may be associated with lower tumor MVD and VEGF expression.
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[Objective]To study the inhibitory effect of the extract of Rhizoma Amorphophallus,we set up the Kunming mices which were vaccinated with H22 tumor as experimental model,and explore the possibility mechanism of them.[Method]We adopted the mices vaccinated with H22 tumor as experimental model to observe the influence of different extracts on the antitumor ratio,and the cooperating antitumor effect together with 5-Fu.Using flow cytometry to detect the effect of petroleum ether extract on apoptotic cells.Using immunohistochemical method to observe the influence of this extract to the expression of Survivin and Bax(Bcl-2 associated protein x).[Result]The extract of petroleum ether of Rhizoma Amorphophalli had inhibitory effect of the growth on mice vaccinated with H22 tumor,and the possible mechanism was the downregulation of Survivin expression and upregulation of Bax expression to induce apoptosis of tumor cell.
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Objective: To study antitumor activity of Zhongjiefeng Injection (Herba sarcandrae) on mice liver cancer Hep A 22 in vivo, in vitro and its toxic reaction in mice with Hep A 22 tumor Methods: The antitumor activity and toxicity of Zhongjiefeng Injection were determined in the mice with transplantable tumor (mice liver cancer Hep A 22), and the antitumor effect of Zhongjiefeng Injection on Hep A 22 cells in vitro were determined with MTT method. Conclusion: The experiment indicates that Zhongjiefeng Injection shows significant antitumon effect increase the indexes of immune organs and amounts of WBC.
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Objective To observe the anti-tumor effect of ethaselen on transplanted liver cancer(H22)and Lewis lung cancer mice.Methods The transplanted liver cancer(H22)and Lewis lung cancer mice models were established.Each genus consisted of 60 mice,which were divided into control,CTX,and three ethaselen dose groups,respectively,with 12 mice in each.Intraperitoneal injection(ip.)of three dosages was performed once a day through the abdominal wall separately,from the second to the eighth day after cancer was transplanted.On the eleventh day,six mice in each group were killed to calculate the tumor inhibition ratio and observe the cell cycle by flow cytometry,and the others were observed for the life extension ratio.Results When ethaselen was given at the dosage of 25,12.5 and 6.3 mg/kg,the tumor inhibition ratio was 52.5%,43.6% and 30.2%,and the life extension ratio was 56.6%,38.7% and 14.2% in the transplanted liver cancer(H22)mice,respectively.The tumor inhibition ratio was 43.8%,29.6% and 18.9%,and the life extension ratio was 47.3%,17.9% and 6.3% in the transplanted Lewis lung cancer mice,respectively.Compared with that of the control,the apoptosis ratio was obviously increased in each group,and there was obviously induced S phase arrest in 25.0 mg/kg(P