AIM: To establish the technique of precision-cut fibrotic liver slice (PCLS) and grope the optimal cultural conditions for researching the liver xenobiotic metabolism in vitro and the drug interaction. METHODS: Complex factors (higher fat diet, alcohol and CCl 4) were used to make the animal model of liver fibrosis. Fibrotic liver slices were prepared and cultivation system was established. Lactate dehydrogenase (LDH) leakage, glutathione S-transferase (GST) activity and 3[4,5-Dimethythiazole-2-yl]-2,5- diphenyltetrazolium bromide (MTT) reduction were chosen as indexes to assess the viability of the slice in different thickness, medium pH and cultural time. RESULTS: Rats were in earlier hepatic fibrosis after administration for 3 weeks. When the thickness of slices was 300 ?m and medium pH was 7.0, the LDH leakage, GST activity and MTT reduction could maintain on a steady level in 6 h. CONCLUSION: A 300 ?m of thickness, 7.0 of medium pH and 6 h of cultural time are the optimal slicing and culturing conditions for fibrotic liver slice.

Analysis of metabolic activities of cytochrome P450 isoenzyme is a crucial index to study drug/toxicant metabolism, drug-drug interaction, polymorphism and et al. Due to this practice, it is important to use the proper probe substrate and to conduct the experiment under optimal conditions. The validation information in literatures on the most common and newest in vitro probe substrates have been reviewed.

Objective To observe the clinical effects of short-term intermittent ganeiclovir treatment on infants with cytomegalovirus (CMV) hepatitis. Methods The clinical data of infants with CMV hepatitis were analyzed retrospectively. The liver functions including total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyltransferase (γ-GT) of the patients in treatment group (85 cases) and control group (37 cases) were collected before and after treatment. Meanwhile, the side effects of ganciclovir during treatment were observed. The measurement data were compared by analysis of variance and numeration data were compared by chi-square test. Results After short-term intermittent ganciclovir treatment in treatment group, TBil level was decreased from (109.1±677.8)μmol/L to (62.9±68.1)μmol/L (F=15.34,P<0.01); ALT level was decreased from (160.2±395.3) U/L to (68.1±56.0) U/L (F=4.73, P<0.05). In control group, TBil level was decreased from (94.9±647.4)μmol/L to (49.2±631.5) μmol/L (F=14.80, P<0.01) ; while ALT level was decreased from (131.6±206.2) U/L to (55.3±631.2) U/L (F=3.50, P=0.067). The readmission rate in control group was significantly higher than that in treatment group (21.6% vs 10.6%). Only one case (0.8%) received three times of intermittent ganciclovir treatment. The longest hospitalization time was six weeks. Conclusions Short-term intermittent ganciclovir treatment may be more suitable for infants with CMV hepatitis. There is no obvious side effect observed during the treatment and the hospitalization time can be shortened.

Objective:To investigate the clinical value of the changes of serum complement,immunoglobulin and inflammatory cytokines in children with mycoplasma pneumonia (MP).Methods:60 cases of children with MP infection were collected as MPP group,60 cases of healthy children as control group.The serum levels of complement (C3,C4) and immunoglobulin (IgA,IgG,IgM) were detected by INA,the serum levels of inflammatory cytokines (IL-8,IL-10,IL-13,TNF-α) were detected by ELISA.Results:①The serum levels of IgM,C3,C4,IL-8,TNF-et and IL-13 in MPP group in acute stage were significantly higher than those in the recovery stage and control group (P＜0.05),the levels of IL-8,TNF-et,IL-13 in the recovery stage were significantly higher than those in control group (P＜0.05);the serum levels of IgA,IL-10 in MPP group in acute stage were significantly lower than those in the recovery stage and control group,and these were still significantly lower than the control group (P＜0.05);the IgG in MPP group in acute stage was not significant difference with control group (P＞0.05),but it in recovery stage was significantly higher than the acute stage and the control group (P＜0.05).②The serum levels of IgM,IgG,C3,C4,IL-8,TNF-et and IL-I 3 in MPP group in severe were significantly higher than those in the mild stage and control group (P＜0.05),and the levels of IgM,IL-8,TNF-αt,IL-13 in the severe stage were significantly higher than those in control group (P＜0.05);the serum levels of IgA,IL-10 in MPP group in severe stage were significantly lower than those in the mild stage and control group,and these in mild stage were still significantly lower than the control group (P＜ 0.05).Conclusion:Children with MP infection have a significant cellular and humoral immune dysfunction,the detection of the changes of serum complement,immunoglobulin and inflammatory cytokines is valuable to assessing the disease staging and degree.

Antineoplastic Agents ; therapeutic use ; Autoimmune Diseases ; pathology ; Benzamides ; Cell Differentiation ; Cell Movement ; Cell Survival ; Eosinophilia ; chemically induced ; pathology ; Eosinophils ; cytology ; physiology ; Helminthiasis ; pathology ; Humans ; Hypereosinophilic Syndrome ; drug therapy ; pathology ; Hypersensitivity ; pathology ; Imatinib Mesylate ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use

Adult ; Female ; Humans ; Male ; Occupational Diseases ; Organometallic Compounds ; poisoning

Air Pollutants, Occupational ; analysis ; Ammonium Sulfate ; analysis ; Chromatography, Ion Exchange ; methods ; Workplace

BACKGROUND:Our previous study has already manifested that Chinese medicine Xiaokening can effectively prevent and treat the early diabetic nephropathy. OBJECTIVE:To compare the effect of rhein and archen on the apoptosis of mesangial cells of rats cultured with high glucose. METHODS:Mesangial cells were stimulated by different concentrations of rhein and archen (20, 40 and 80 μmol/L). Morphology of apoptotic cells was observed by hematoxylin-eosin staining. Karyon apoptosis was examined by fluorescence staining of DAPI. cellapoptosis rate was detected by flow cytometry. RESULTS AND CONCLUSION:The outcome of hematoxylin-eosin staining and DAPI staining indicated that the effect of rhein was much stronger than that of archen on the apoptosis of mesangial cells in the rats cultured with high glucose. Comparison of the apoptosis rate also indicated that the rhein had a stronger effect on the earlier and the later stage apoptotic rate of mesangial cells than archen. Both rhein and archen with different concentrations can induce apoptosis of mesangial cells, but the effect of rhein is much stronger.

Objective To explore the effects of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] on memory CD4+T cells of focal proliferative IgA nephropathy (IgAN)patients.Methods (1) Total of twenty incipient focal proliferative IgAN patients (Lee classification:Ⅲ level) were chosen as IgAN group and 20 healthy volunteers were chosen as healthy control group.The level of serum 1,25(OH)2D3 was measured by radioimmunoassay (RIA).Peripheral blood mononuclear cells (PBMCs) were separated by the method of Ficoll density gradient centrifugation and were stimulated with anti-CD3/anti-CD28 in the absence or presence of various concentrations of 1,25(OH)2D3,Dexamethasone(DEX) and 1,25(OH)2D3and DEX combined.PBMCs were cultured for 72 hours and the levels of IFN-γ,IL-4,IL-17A,Foxp3 were measured by flow cytometry(FCM),standing for the levels of Th1,Th2,Th17,Treg.(2) IgAN group was divided into two subgroups (proteinuria ＜ 1 g/24 h subgroup,proteinuria≥ 1 g/24 h subgroup),then the serum levels of 1,25(OH)2D3,IFN-γ,IL-4,IL-17A,Foxp3 were compared.Results Compared with healthy control group,serum 1,25(OH)2D3 level of IgAN group was significantly lower (P ＜ 0.05).Serum 1,25(OH)2D3 level in proteinuria≥ 1 g/24 h subgroup was significantly lower than proteinuria ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The level in proteinuria ＜ 1 g/24 h subgroup was lower than healthy control group,but the difference was not statistically significant (P ＞ 0.05).(2) The levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4,IL-17A/Foxp3 in IgAN group increased significantly compared with healthy control group (all P ＜ 0.05),and the level of Foxp3 decreased significantly (P ＜ 0.05).The level of IL-4 also increased,but the difference was not statistically significant (P ＞ 0.05).The levels of IFN-γand IL-17A and the ratio of IL-17A/Foxp3 in proteinuria≥ 1 g/24 h subgroup increased significantly,and the level of Foxp3 decreased significantly,compared with urinary protein ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The ratio of IFN-γ/IL-4 in proteinuria≥1 g/24 h subgroup and proteinuria ＜ 1 g/24 h subgroup all increased,compared with healthy control group,and the ratio in proteinuria≥ 1 g/24 h subgroup increased significantly (P ＜ 0.05).There was no significant difference in the level of IL-4 among all groups.(3) After treatment with 1,25(OH)2D3,the levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3 decreased significantly,and the level of Foxp3 increased significantly (P ＜ 0.05),and these effects were more obvious as the increase of the drug concentration.The level of IL-4 did not change significantly.The combination of 1,25(OH)2D3 and DEX had a synergistic inhibition on the production of IFN-γ,IL-4,IL-17A,and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3,and had a synergistic promotion on the production of Foxp3.Conclusions There is a certain extent of vitamin D deficiency in focal proliferative IgAN patients,which may be associated with the severity of proteinuria.The disorder of immunomodulatory effects of memory CD4+ T cell might exist in the patients of focal proliferative IgAN.1α,25-dihydroxyvitamin D3 might have beneficial effects on the immunoregulation of memory CD4+T cells of focal proliferative IgAN patients.

Objective To construct expression vectors of human ribosomal protein S 3(RPS3) and RPS3Ser209 mutant in orcler to investigate the effect of RPS3Ser209 mutant on NF-κB signaling pathway and DNA binding capacity .Methods The vector RPS3-myc was amplified by polymerase chain reaction ( PCR) from the human liver cDNA and subcloned into pcDNA-3.1myc-HisB.RPS3S209A represented mutant RPS3 expression vectors, in which the designated amino acid was mutated to an alanine residue .Dual luciferase reporter gene assay was used to detect the NF-κB transcription activity in HEK293 cells,immunofluorescence to detect RPS3 location, and EMSA to examine NF-κB DNA-binding activity.Results The expression vectors of RPS3-myc and RPS3S209A-myc were constructed.Compared with wild-type RPS3,the nucleus translocation, transactivation activity of NF-κB and DNA binding ability of RPS3S290A were reduced significantly .Conclu-sion The impact of RPS3 on NF-κB signaling pathway depend on its serine 209.