OBJECTIVETo study the effect and mechanism of Ganbi decoction (GBD) in treating patients with antituberculotic agent caused liver injury (ATB-LI).

METHODSOne hundred and twenty-eight patients with ATB-LI were randomly assigned to the treated group (n = 66) and the control group (n = 62) with the envelop method. Meanwhile, 60 healthy persons were selected as the healthy control group. The treated group was treated by GBD one dose every day with the constituents modified depending on patients' symptoms, and the control group was treated with glucuronolactone tablets and inosine injection. One week was taken as one treatment course. The changes of clinical syndromes, physical signs, T-lymphycyte sub-groups and serum level of nitric oxide (NO) were observed before and after treatment and the recovery time of liver function was recorded. The outcome was compared with that in the healthy control group.

RESULTSIn the treated group, 28 patients (42.4%) were cured, 30 (45.5%) improved and 8 (12.1%) ineffectively cured, the total effective rate being 87.9% (58/66). In the control group, 17 patients (27.4%) were cured, 24 (38.7%) improved, and 21 (33.9%) ineffectively cured, the total effective rate being 66.1% (41/62). The total effective rate in the treated group was significantly higher than that in the control group (P < 0.05). Liver function was improved in both groups, recovery time in the treated group was 12.0 +/- 7.0 days, which was significantly shorter than that in the control group (16.0 +/- 8.0 days), showing significant difference between the two groups (P < 0.05). The levels of CD3, CD4 and CD8 were significantly higher and level of NO significantly lower in the two groups of patients than those in the healthy control group (P < 0.05), but these parameters were improved more significantly in the treated group after treatment, when compared with those before treatment or with those in the control group, all showing significant difference (P < 0.05).

CONCLUSIONGBD could prevent ATB-LI, and its mechanism could be by way of reducing NO production induced by endotoxin of macrophage and stimulating the proliferation of T-lymphycyte to elevate immunity.

Adult ; Aged ; Antitubercular Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucuronates ; therapeutic use ; Humans ; Inosine ; therapeutic use ; Liver Diseases ; drug therapy ; Liver Function Tests ; Male ; Middle Aged ; Nitric Oxide ; blood ; T-Lymphocyte Subsets ; Treatment Outcome

To investigate the delivery mechanism of micro-porous osmotic pump tablets ( MPOP), taking tramadol hydrochloride ( TR) as the model drug, tramadol hydrochloride micro-porous osmotic pump tablets (TR MPOP) were prepared with compressible starch as diluent, cellulose acetate as coating material, polyethylene glycol 400 as pore-forming agents. The equilibrium solubility and osmolality of TR were determined. The effects of fillers in tablet cores, coating levels, and osmotic pressures of release media on expansion behavior of preparations were described. The influences of the category, osmolality, and pH value of release media, release methods, and release conditions on release curves of tablets were evaluated. Based on several models, the delivery pattern of TR MPOP was fitted. The equilibrium solubility in water and osmolality of TR were (775.8 +/- 17.7) g x L(-1) and 4.036 Osmol x kg(-1), respectively. During the drug-release period, it was observed that the tablets expanded markedly in response to the expansion characteristics of compressible starch and the osmotic pressure difference across the membrane. When osmotic pressure of release media increased, the significant change of the equilibrium solubility of TR was not found, but the release rates of TR MPOP decreased significantly. The delivery rate was not influenced by the pH of release mediums, dissolution methods and paddle stirring rates. The drug release profile conformed to the model of zero order in 8 h. The pore-forming agents were dissolved in release medium, which caused micro-pores. The expansion of tablets made the size of micropores bigger, and then the drug-releasing pores were obtained. It was proved that the drivers of drug delivering from TR MPOP were mainly the difference of osmotic pressure, and secondly the difference of solubility. TR MPOP were the controlled-release preparation.
Analgesics, Opioid ; administration & dosage ; chemistry ; Cellulose ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; methods ; Drug Stability ; Osmosis ; Osmotic Pressure ; Polyethylene Glycols ; chemistry ; Porosity ; Solubility ; Starch ; chemistry ; Tablets, Enteric-Coated ; Technology, Pharmaceutical ; methods ; Tramadol ; administration & dosage ; chemistry ; pharmacokinetics

OBJECTIVETo explore the effects of HBsAg pulsed dendritic vaccination on anti-HBs production in immunosuppressed rats after liver transplantation (LT).

METHODSBrown-Norway liver allografts were transplanted into Lewis recipients. The transplanted Lewis rats were injected with EK506 (2 mg/kg) and randomly divided into two groups: rats in HBsAg-DCs group (n = 15) were intraperitoneally injected with HBsAg pulsed DCs at 14 d and 28 d after LT, and rats in the HBsAg group (n = 15) were injected with HBsAg (200 mul) once a week for 12 weeks. Rats without any immunosuppressive treatment after LT served as controls (n = 5). IL-2 and IFN-gamma mRNA expression in spleen were analyzed by RT-PCR, serum IL-2, IFN-gamma and anti-HBs were detected by ELISA.

RESULTSHigh dose of FK506 resulted in the immunosuppressed in LT rats, as evident by low production of IL-2 and IFN-gamma, and without liver rejection compared to rats in the control group. HBsAg-DCs induced high titer of anti-HBs antibody, however, titer of anti-HBs were seldom detectable in the HBsAg group at 1, 2 and 3 mouth after vaccination.

CONCLUSIONThe capacity of HBsAg-DCs to induce anti-HBs in immunosuppressed rats suggested that DC vaccine may prevent HBV recurrence in liver transplanted patients.

Adjuvants, Immunologic ; pharmacology ; Animals ; Cytokines ; blood ; genetics ; metabolism ; Dendritic Cells ; immunology ; Disease Models, Animal ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; Immunosuppression ; Immunosuppressive Agents ; administration & dosage ; Liver Transplantation ; immunology ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Secondary Prevention ; Spleen ; immunology ; metabolism

OBJECTIVETo study the mechanisms of pseudolaric acid B-induced apoptosis on A375-S2 cells.

METHODMTT, fluorescence microscope observation, DNA agarose gel electrophoresis and Western blot analysis wereused.

RESULTPseudolaric acid Binduces A375-S2 cell apoptosis in a time and dose-dependent manner. Apoptotic bodies and DNA ladder were observed in 5 micromol x L(-1) pseudolaric acid B-treated A375-S2 cells for 36 h. The expression of Bcl-2, Bcl-xL and ICAD was reduced time dependently, whereas the expression of Bax was increased.

CONCLUSIONThe major cause of pseudolaric acid B induced cytotoxicity on A375-S2 cells was apoptosis. Mitochondria proteins and ICAD might be involved in the apoptotic pathways of pseudolaric acid B-treated A375-S2 cells.

Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Cell Line, Tumor ; Diterpenes ; isolation & purification ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Melanoma ; metabolism ; pathology ; Pinaceae ; chemistry ; Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; bcl-X Protein

OBJECTIVETo establish the quality standard of PsL injections containing mainly 5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid).

METHODThe identification of PsL was performed by thin-layer chromatography, and the content was determined by HPLC. The column was Hypersil C18 (4.6 mm x 250 mm, 5 microm), the mobile phase was the mixture of methane-water-acitic acid (55:45: 0.045) with a flow rate of 1.0 mL x min(-1), the detective wavelength was 254 nm, and the column temperature was maintained at 35 degrees C. The pH value and K+ content of the three batchs injection were determined with pH meter and flame photometric meter, and the contents of tannin, protein, oxalic acid salt and heavy metals were detected by deferent methods.

RESULTThe TLC method was suitable for the identification of PsL5F. The linearity for 5F was obtained over the range of 30-240 microg x mL(-1) (r = 0.999 8), the average recovery of 5F was 99.8%. The injections were of pH value range from 7.80 to 8.20, K+ contents less than 10 mmol x L(-1), and the contents of tannin, protein, oxalic acid salt and heavy metals were qualified with the Chinese pharmacopoeia, respectively.

CONCLUSIONIt's sensitive and reliable that can be used as quality control methods of PsL5F injections.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Diterpenes ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Injections ; Reproducibility of Results

OBJECTIVETo study the mechanisms of capsaicin-induced apoptosis of human melanoma A375-S2 cells.

METHODSMTT assay, fluorescence microscopy, DNA agarose gel electrophoresis, flow cytometry and Western blot analysis were carried out to assess the morphological and biochemical changes of A375-S2 cells after capsaicin treatment.

RESULTSCapsaicin induced A375-S2 cell death in a time- and dose-dependent manner. Sub-diploid peak was seen at 24 h after 250 micromol/L capsaicin treatment, and apoptotic bodies and DNA ladder were observed at 36 h after capsaicin treatment. The expression of inhibitor of caspase activated DNase (ICAD) was reduced with the lapse of time.

CONCLUSIONCapsaicin induces A375-S2 cell apoptosis and down-regulation of ICAD contributes to this process.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; biosynthesis ; Capsaicin ; pharmacology ; Caspases ; metabolism ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Humans ; Melanoma ; pathology ; Signal Transduction ; Skin Neoplasms ; pathology

Objective To study Caveolin-1,EGFR expression in bladder transitional call carcinoma and their prognostic value. Methods Immunohistochemical method was used to detect Caveolin-1,EGFR in 89 cases.of bladder transitional call carcinoma.Results In 89 cases,the percentage of abnormal Caveolin-1 and EGFR expression were 37.1% and 50.6 % respectively.Significant change was observed in different grade case,P

OBJECTIVETo study the distribution of human immunodeficiency virus-1 (HIV-1) genotypes in Hubei province.

METHODSEpidemiological survey was carried out to HIV-1 carriers who were identified in Hubei province. HIV-1 env V3-V4, gag P17/24 and the first exon of tat region were amplified by nested-polymerase chain reaction(nPCR) .The sequences were determined, and phylogenetic analyses were then performed.

RESULTS4 HIV-1 strains or circulating recombinant forms (CRFs) were identified in Hubei province with subtype B' the predominant which covered 5 kinds of populations including former blood donors, blood receivers, spouses of the infected people, sex workers and their clients, homosexuals, mainly distributed in the areas with many former blood donors. CRF08-BC and CRF01-AE were found distributed in economically more developed cities or southern area of the province, and the major transmission routes was through sexual contact. Only 1 patient, an injecting drug user, was identified having subtype C.

CONCLUSIONSubtype B' was the main epidemic subtypes in Hubei province while CRF08-BC, CRF01-AE and subtype C were also circulating in the province, indicating the transmission of the disease might to become more complex.

China ; epidemiology ; HIV Infections ; epidemiology ; HIV-1 ; classification ; Humans ; Molecular Epidemiology ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, RNA

Objective To investigate the expression of cyclin dependent kinases 5(CDK5)in the temporal lobes of the epilepsy patients and to explore the possible roles of CDK5 in the pathogenesis of refractory epilepsy.Methods The brain tissues of intractable epilepsy(IE)were studied by fluorescence quantative polymerase chain reaction(FQ-PCR)for CDK5 mRNA,while immunohistochemistry and Western blot were used to study the protein expression.Nonepileptogenic control brain tissues were used for comparison.Results FQ-PCR analysis showed that the expression of CDK5 mRNA in epilepsy patients was significant higher than those in the control group.And immunohistochemistry showed that the protein mainly existed in the neuron and glial.At the 35000 relative molecular mass,Western blot could been seen that there is a limpid strap.The optical density of CDK5 in IE(temporal lobe 1.4293?0.1839,hippocampus 2.0733?0.4738)was significantly higher than that in the control(temporal lobe 0.9680?0.4147, hippocampus 1.4030?0.6160,P

OBJECTIVETo explore the inhibitory effects on the activation of endotoxin-induced Kupffer cells (KCs) through short hairpin RNA (shRNA) targeting interleukin-1 receptor associated kinase-4 (IRAK-4) gene.

METHODSTwo effective transfection shRNA plasmid (pSIIRAK-4-A, pSIIRAK-4-B) and one invalidated plasmids (pSIIRAK-4-C) targeting IRAK-4 gene were constructed. The isolated mouse KCs were divided into three groups: the normal control group, the RNAi control group (pSIIRAK-4-C) and the RNAi effective group (pSIIRAK-4-A, pSIIRAK-4-B). Then KCs were stimulated with 0.1 microg/ml lipopolysaccharide (LPS) after 24 h transfection with the constructed plasmid. The expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot at 6 h after LPS stimulation, and the activities of NF-kappaB in KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h.

RESULTSThe level of IRAK-4, the activities of NF-kappaB and the TNF-alpha level in the RNAi effective group were evidently lower than those in normal and RNAi control groups (P < 0.01) at 1 h, 3 h, and 6 h. Especially, the pSIIRAK-4-A group in which the changes of the above indices were of no difference (P > 0.05), had better inhibited effects than that of the pSIIRAK-4-B group (P < 0.01).

CONCLUSIONThe shRNA targeting IRAK-4 gene could effectively inhibit the activation of endotoxin-induced KCs.

Animals ; Endotoxins ; Interleukin-1 Receptor-Associated Kinases ; genetics ; metabolism ; Kupffer Cells ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Small Interfering ; genetics ; Signal Transduction ; physiology