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OBJECTIVE：To prelimi narily investigate the possible mechanism of orazamide to prevent anti-tuberculosis drug-induced liver injury （ATB-DILI）. METHODS ：A total of 60 Kunming mice were randomly divided into blank group ，model group，positive control group [diammonium glycyrrhizinate 60 mg/（kg·d）]，orazamide low-dose ，medium-dose and high-dose groups [ 80，160，320 mg/（kg·d）]，with 10 mice in each group. Except for blank group ，other groups were given isoniazid [ 75 mg/（kg·d）]+rifampicin [ 75 mg/（kg·d）] for 14 days intragastrically to induce ATB-DILI model. At the same time ，administration groups were given relevant medicine intragastrically ，blank group and model group were given normal saline intragastrically. The administration volume was 20 mL/（kg·d），once a day ，for consecutive 14 days. The general conditions of the mice were observed and recorded every day ，such as growth and development ，mental and diet state. After last medication ，liver index was calculated ， and HE staining was adopted to observe pathological changes of liver tissue of mice. The positive expression of high mobility group protein B 1 （HMGB1） and NF-κ B in liver tissue were detected by streptavidin biotin-peroxidase complex （SABC） immuno- histochemistry. The serum levels of liver function indexes in serum ，the protein expression of advanced glycation end product receptor（RAGE）and TNF-α in liver tissue were detected by ELISA. RESULTS：Compared with blank group ，the growth and development of mice in the model group were slow ，and their appetite and spirit were poor. The liver index ，serum levels of TBIL ， DBIL，ALT，AST，ALP，TBA and γ-GT were increased significantly （P＜0.05）. Structural disorder of liver lobules ，degeneration and necrosis of liver cells and inflammatory cell infiltration were observed. The expression of HMGB 1，NF-κB，RAGE and TNF-α in liver tissue were elevated significantly （P＜0.05）. Compared with model group ，the general condition of mice were all improved to different extents in orazamide low-dose ，medium-dose and high-dose groups ，positive control group ，while liver index and above serum indexes were all decreased significantly （P＜0.05）. The pathological changes of liver tissue were all improved to different extents ，while the protein expression of HMGB 1，NF-κB，RAGE and TNF-α were all decreased significantly（P＜0.05）. The improvement of above indexes in orazamide high-dose group were all significantly better than orazamide low-dose and medium-dose groups （P＜0.05）；the levels of ALP and TBA in orazamide high-dose group were significantly lower than positive control group （P＜0.05）. CONCLUSIONS ：Orazamide can prevent ATB-DILI induced by isoniazid combined with rifampicin in mice，the mechanism of which may be associated with down-regulating the protein expression of HMGB 1 and RAGE in liver tissue and inhibiting the secretion of inflammatory factors.
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Objective To investigate the changes and clinical significance of helper T cells (Th)22,Th17,Th1 and regulatory T cells (Treg) in the peripheral blood before and after antiretroviral treatment (ART) of human immunodeficiency virus (HIV)-1 infected patients.Methods Forty HIV-infected patients were recruited into this study,and 30 healthy subjects were recruited as controls.Peripheral blood of the patients was collected at baseline and after 3 months of ART treatment.The frequencies of Th22,Th17,Th1 and Treg were detected by flow cytometry.Tests for homogeneity of variance and paired t test for comparison was adopted.Spearman rank test was used for correlation analysis.Results The frequencies of peripheral Th22,Th17,Th1 and Treg from HIV-infected patients before treatment were significantly decreased compared to the healthy controls ([0.59± 0.47] ％ vs [1.65 ± 0.56] ％ [t =8.544,P＜0.01],[4.46±1.84]％ vs [6.98±1.86]％[t=5.619,P＜0.01],and [16.75±6.72]％ vs [22.77±6.87]％; [t=5.311,P＜0.01].The frequencies of peripheral Th22 and Th17 after 3 months of treatment were significantly higher than those at baseline ([1.60± 1.10] ％ [t=5.268,P＜0.01] and [6.33±2.64]％ [t=3.663,P＜0.01] and no difference from those of healthy controls (t=1.783 and 1.143,respectively; both P＞0.05).However,the Th1 frequency showed no difference compared to the baseline.The frequency of Treg in the HIV infected patients was significantly increased compared with the healthy controls ([10.76±3.76]％ vs [7.01±1.88]％,t=5.003,P＜0.01).However,it gradually decreased along with the ART treatment ([9.22±2.56]％ after 2 months; [8.57± 2.36]％ after 3 months),which was still higher than that of healthy controls (t=2.984,P=0.004).The ratios of Th22/Treg,Th17/Treg and Th1/Treg of the HIV-infected patients were significantly decreased in comparison with the healthy controls (0.05±0.03 vs 0.25±0.10,t=11.69,P＜0.01; 0.46 ± 0.27 vs 1.07±0.42,t=7.728,P＜0.01; 1.56±0.89 vs 3.37± 1.02,t=7.052,P＜0.01),and those were significantly increased after 3 months of treatment (0.17±0.10[t=6.852,P＜0.01],0.81±0.46[t=4.253,P＜0.01] and 2.31±1.27[t=3.030,P＜0.01]).Correlation analysis showed that the ratio of Th17/Treg of the HIV-infected patients was positively correlated with the peripheral CD4+ T cell count (r=0.312 5,P=0.049 6),and negatively correlated with HIV RNA viral load (r=-0.474 7,P=0.002 0).The ratio of Th1/Treg of the HIV-infected patients was positively correlated with the peripheral CD4+ T cell count (r=0.333 5,P=0.035 5).Conclusions Th22,Th17,Th1 and Treg cells in the peripheral blood of HIV-infected patients are closely related to CD4+ T cell count.ART can partially recover immune imbalance,and help to rebuild immune function of HIV-infected patients.
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Objective To investigate the inhibitive activities of hepatitis B immunoglobulin(HBIG)poly(butylcynaoacrylate)nanoparticles(HBIG-PBCA-NP)to hepatitis B surface antigen(HBsAg)and hepatitis B virus(HBV)DNA secretions using HBV infected cell model in vitro.Methods HepG 2.2.15 cells were cultured with media containing HBIG-PBCA-NP or HBIG for several days,or cultured with HBIG-PBC-NP and HBIG for 2 days and without HBIG-PBCA-NP and HBIG from day 3.The supernatants at different time points were collected for quantitative detection of HBsAg and HBV DNA.The comparisons between groups were done by variance analysis.Resalts Secretions of HBsAg and HBV DNA in supernatants of HepG2.2.15 cultured with 0.1-10.0 IU/mL of HBIG-PBCA-NP and HBIG were inhibited significantly compared with control group.HBsAg titers and HBV DNA levels in supernatants of HBIG-PBCA-NP group and HBIG group cultured with media without HBIG-PBCA-NP and HBIG kept decreasing at day 5 and 7,then rebounded at day 9 and 11.HBsAg titera in supernatants of 0.1,1.0,5.0 IU/mL HBIG-PBCA-NP group were all significantly different from those in HBIG group at day 9[(31.31±1.98)μg/L vs(40.62±2.99)μg/L,(23.79±1.31)μg/L vs(36.51±2.12)μg/L,(19.91±1.74)μg/L vs(33.03±1.65)μg/L;F=412.24,P＜0.01].Couclusion HBIG-PBCA-NP can inhibit secretions of HgsAg and HBV DNA in vitro,which is more effective than HBIG.
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OBJECTIVE:To investigate the anti-HBV activities of deoxynojirimycin(DNJ)derivatives N—benzyl—1—DNJ(P-DNJ)and N—nonyl—1—DNJ(NN-DNJ)in vitro.METHODS:Human hepatoma carcinoma cell lines HepG2 2,2,15,which were transfected from HBV DNA,were taken as target cells,cells were cultured with different concentrations of test drugs, with HBsAg,HBeAg and HBV DNA in the cultured supernatant determined by time-resolved immunofluorometric assay and fluorescent quantitation PCR assay on day 6th and 10th day,meanwhile,with the cytotoxicity of DNJ derivatives determined by MTT assay.RESULTS:No cytotoxic effects was noted when the test concentration of P-DNJ and NN-DNJ was within 5～125?g?mL-1,HBsAg and HBeAg levels and HBV DNA secretion decreased significantly at a concentration of 125?g?mL-1.CONCLUSION:P-DNJ and NN-DNJ showed anti-HBV activity in the in vitro cell culture experiment.