Animals ; Blotting, Western ; Cell Differentiation ; drug effects ; Embryonic Stem Cells ; cytology ; enzymology ; Isoenzymes ; analysis ; Mice ; Neurons ; enzymology ; Protein Kinase C ; analysis ; Tretinoin ; pharmacology

OBJECTIVETo investigate the effects of benzo[a]pyrene (B[a]P) exposure on the behaviors and hippocampal oxidative stress and ATPase in rats and the molecular mechanism of neurobehavioral toxicity of B[a]P.

METHODSA total of 120 male SD rats (21 days old) were randomly and equally assigned to five groups: blank control group, vegetable oil (solvent control) group, and 2.5, 5, and 10 mg/kg B[a]P exposure groups. The rats in B[a]P exposure groups were injected intraperitoneally with B[a]P once a day for 4 consecutive weeks. Then, Morris water maze and shuttle box were used to evaluate the learning and memory abilities of rats; colorimetric assay was used to measure the activities of superoxide dismutase (SOD), Na(+)/K(+)-ATPase, and Ca(2+)/Mg(2+)-ATPase and the content of malonaldehyde (MDA) in the hippocampus; the concentration of Ca(2+) in the hippocampus was measured by fluorescent labeling.

RESULTSCompared with the blank control group and solvent control group, the B[a]P exposure groups exhibited significant increases in escape latency, active avoidance response latency, and passive avoidance response latency and significant decreases in number of platform crossings and active avoidance response frequency in the last test (P < 0.05 for all comparisons), with a dose-effect relationship. In addition, the B[a]P exposure groups had significantly lower activities of SOD, Na(+)/K(+)-AT-Pase, and Ca(2+)/Mg(2+)-ATPase and significantly higher MDA level and Ca(2+) concentration than the blank control group and solvent control group (P < 0.05 for all comparisons), with a dose-effect relationship.

CONCLUSIONThe neurobehavioral toxicity of B[a]P may be related to increased oxidative stress and decreased activities of Na(+)/K(+)-ATPase and Ca(2+)/Mg(2+)-ATPase in the hippocampus of rats.

Animals ; Benzo(a)pyrene ; toxicity ; Ca(2+) Mg(2+)-ATPase ; metabolism ; Hippocampus ; drug effects ; metabolism ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

Objective To investigate the relationship between interleukin 10 (IL-10) -592 (rs1800872) and -819 (rs1800871) promoter genetic polymorphisms and the susceptibility of antituberculosis drug-induced liver injury (ADLI). Methods A case-control study was conducted. Epidemiology survey data and peripheral blood samples were obtained from the patients. Two IL-10 gene polymorphisms (-592 A/C and 819 C/T) were genotyped with PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) in Chinese Han ADLI subjects (n=180) and sex matched by frequency matching in control subjects (n=180). Results No significant differences in genotypes of IL-10 -592 site and IL-10 -819 site between ADLI group and that of the control group were noticed (all P>0.05). The mutant alleles -592 C of IL-10 gene polymorphism was significantly higher in ADLI subjects compared to controls, and in dominant model, the frequency of CC+AC genotype was 1.62 higher among the cases than controls (all P<0.05). Significant difference in allele -819 C/T between the ADLI group and the control group were not found (P=0.190). The polymorphisms at -819 C/T and -592 A/C variants of IL-10 gene were found to be good linkage disequilibrium. The CC haplotype represent genetic risk factor (OR=1.37, 95% CI: 1.02-1.85) and CA haplotype represent genetic protect factor (OR=0.49, 95% CI: 0.34-0.70) for ADLI in the subjects. Conclusions The polymorphisms in IL-10 gene -592 A/C and -819 C/T are associated with ADLI.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Middle Aged ; Mutation ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spastic Paraplegia, Hereditary ; genetics ; Young Adult

OBJECTIVETo study the specific cellular immunoresponse of peripheral blood lymphocytes in the chronic hepatitis B patients treated with different doses of recombinant hepatitis B vaccine.

METHODSSeventy-two chronic hepatitis B patients who did not use any anti-HBV drugs within 6 months were randomized into 3 groups (90 micrograms, 60 micrograms, and placebo) in a ratio of 1:1:1. The patients in different groups were treated with different doses of recombinant hepatitis B vaccine in combination with IFN alpha 1b 50 micrograms with 3 times a week for 24 weeks. All patients were followed up for 24 weeks (W24). HBV DNA, HBeAg and liver functions were detected at different time points, and the number of cells that secrete IFN-gamma were detected by ELISPOT.

RESULTSThere were no significant difference in ELISPOT positive ratio among the 3 groups on baseline detection. At W24, 12 cases, 12 cases, and 7 cases showed ELISPOT positive in the group of 90 micrograms, 60 micrograms, and placebo. The proportion of patients who were ELISPOT positive was higher in the groups treated with recombinant hepatitis B vaccine (including the dose of 90 micrograms and 60 micrograms) than that in the placebo group (P=0.0446). HBV DNA turned negative in 6/24 of the patients treated with recombinant hepatitis B vaccine (at both the doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg became negative in 7/24 of them. In the placebo group, none of the patients showed undetectable HBV DNA, HBeAg/Anti-HBe seroconversion or HBeAg disappearance. At the 24W of follow up, in the patients who were ELISPOT positive, HBV DNA became undetectable in 4 of the patients treated with recombinant hepatitis B vaccine (at doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg disappearance were found in 9 of the cases. In the placebo group, none of the cases showed undetectable HBV DNA, and only 1 case had HBeAg/Anti-HBe seroconversion.

CONCLUSIONThe recombinant hepatitis B vaccine may increase the function of specific T lymphocytes in patients with chronic hepatitis B. There were no significant differences between the patients treated with the dose of 90 micrograms and 60 micrograms hepatitis B vaccine.

Adult ; DNA, Viral ; blood ; Female ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Recombinant Proteins ; immunology ; T-Lymphocytes ; immunology ; Vaccines, Synthetic ; immunology

Human parathyroid hormone (hPTH) was highly expressed in Escherichia coli by inserted the synthesized whole hPTH cDNA into the vectors pBV220 and pET22b. After expression and disruption, the purified product was acquired through cation exchange chromatography and reverse phase chromatography. From the results of N-terminal sequencing and MALDI-TOF-MS analysis the recombiant prtein was indentified as intact hPTH. In in vitro Bioassays the recombinant hPTH stimulated adenylate cyclase as the standard did. In ovariectomized rats the recombinant hPTH markedly increased the femoral bone mass and bone mineral density.
Amino Acid Sequence ; Animals ; Base Sequence ; Bone Density ; drug effects ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Molecular Sequence Data ; Ovariectomy ; Parathyroid Hormone ; chemistry ; genetics ; metabolism ; pharmacology ; Rats ; Rats, Wistar ; Sequence Alignment ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo study PD-1 and PD-L1 expressions during 24 weeks telbivudine antiviral treatment in patients with chronic hepatitis B (CHB) and to explore the relationship between PD-1 expression and HBeAg/HBeAb seroconversion.

METHODSTen CHB cases with HLA-A2 and HBeAg positive were treated with telbivudine 600 mg/d orally for 24 weeks. Fresh blood samples were collected at week 0, 12 and 24 after treatment. HBV-specific CD8+ T cells were expanded in vitro. Cell culture medium were collected for interferon gamma (IFNgamma) detection. Flow cytometry was used to detect the HLA-A type, PD-1, PD-L1 and HBV specific CD8+ T cells. The expressions of PD-1 and PD-L1, the counts of HBV-specific CD8+ T cells in circulating CD8+ lymphocytes, and IFNgamma concentration in culture medium were evaluated during antiviral treatment.

RESULTSAt week 0, 12 and 24 after telbivudine treatment, 7 of 10 patients were HBV DNA undetectable, 2 were HBeAg seroconversion and 2 were HBeAg lose but anti-HBe negative. The frequency of PD-1-positive PBMCs were 52.1%+/-17.0%, 39.1%+/-18.2% and 23.4%+/-16.3% (week 24 vs week 0, P < 0.01) respectively; PD-L1 positive PBMCs were 45.6%+/-15.4%, 34.6%+/-16.2% and 20.9%+/-9.5% respectively(week 24 vs week 0, P < 0.01; week 24 vs week 12, P < 0.05). The frequency of PD-1-positive CD8+ T cells were 76.2%+/-10.4%, 66.5%+/-15.4% and 49.5%+/-25.3% respectively (week 24 vs week 0, P < 0.01; week 12 vs week 0, P < 0.05; week 24 vs week 12, P < 0.05); HBV-specific CD8 cells were 1.3%+/-0.5%, 1.5%+/-1.0% and 2.2%+/-1.5%; IFNgamma levels in cell culture medium were (91.7+/-82.1) pg/ml, (99.4+/-93.5) pg/ml and (109.5+/-86.6) pg/ml. A remarkable decrease of PD-1 and PD-L1 expressions and increase of HBV-specific CD8+ T cells were observed in patients who had HBeAg/HBeAb seroconversion at week 24.

CONCLUSIONSDirect suppression of HBV replication by telbivudine in CHB patients can decrease PD-1 and PD-L1 expressions and restore HBV-specific CD8+T cells. The relationship between the changes of PD-1 expression and HBeAg/HBeAb seroconversion during antiviral therapy in HBeAg-positive patients need to confirm by future study.

Adult ; Antiviral Agents ; therapeutic use ; B7-H1 Antigen ; metabolism ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; metabolism ; Humans ; Male ; Nucleosides ; therapeutic use ; Programmed Cell Death 1 Receptor ; metabolism ; Pyrimidinones ; therapeutic use ; Thymidine ; analogs & derivatives ; Young Adult

OBJECTIVETo investigate the effect and mechanism of BSDW on the model of allergic rhinitis and the model of guinea pigs by histamine shocking in guinea pigs.

METHODUsing the model of allergic rhinitis in guinea pigs caused by 10% TDI, we observed the effect of BSDW on physiological and pathological symptoms of allergic rhinitis in guinea pigs, the effect of the levels of serum IgE and serum and nasal histamine. Using the model of guinea pigs by histamine shocking, we observed the effect of BSDW on physiological symptoms in guinea pigs.

RESULTBSDW significantly relieved the pathological symptoms of allergic rhinitis in guinea pigs, alleviated the hyperplasia of columnar epithelium, decreased the number of monocyte and eosinocyte compared with the model group. It also reduced the levels of serum IgE, and decreased the release of serum and nasal histamine. BSDW significantly prolonged the occurent time of gasping, eclampsia and death caused by shock, reduced the times of gasping in the model of guinea pigs by histamine shocking.

CONCLUSIONBSDW has significant effect against allergy. The mechanism relates to its effects of decreasing the levels of serum IgE and inhibiting the release of serum and nasal histamine.

Administration, Intranasal ; Animals ; Anti-Allergic Agents ; pharmacology ; Asarum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Histamine ; blood ; Immunoglobulin E ; blood ; Lamiaceae ; chemistry ; Male ; Nasal Mucosa ; immunology ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; immunology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

OBJECTIVETo investigate the proliferation of mouse splenic lymphocytes after shRNA mediated BTLA gene silence.

METHODSThree specific shRNAs and one nonspecific shRNA (scrambled) were ligated to pSilencer3.1-H1-neo plasmid and then subcloned into lentiviral vector pLB. Recombinant viral particles were harvested post-transduction to 293T cells. Mouse lymphocytes were infected with viral supernatant after 24 h incubation and continuously cultured till 4 days. Expression of eGFP was detected by fluorescence microscopy, efficiency of infection and expression of BTLA on lymphocyte cell by FCM. CCK-8 assay was used to detect the proliferation of lymphocytes.

RESULTSLentiviral expression vectors pLB-shRNA/BTLA were successfully generated. The lentiviral particles were correctly packaged. Expression of BTLA protein in specific shRNA group was significantly decreased comparing to those in control group. O.D. value at A(450) of lymphocytes stimulated by anti-CD3 antibody showed significant difference compared with normal BTLA group (P < 0.05), while there was no difference between ConA stimulated group and control (P > 0.05).

CONCLUSIONGene-specific shRNA can knockdown the expression of BTLA. The proliferation of lymphocytes stimulated by anti-CD3 antibody after RNAi demonstrates significant enhancement as compared to the unstimulated lymphocytes, while stimulated by ConA showed no difference compared to normal lymphocytes.

Animals ; Genetic Vectors ; Lentivirus ; genetics ; Lymphocytes ; drug effects ; Mice ; Plasmids ; RNA, Small Interfering ; genetics ; Transfection

Throat swabs collected from patients whose serum was measles IgM negative and rubella IgM positive during 2001-2011 were used to conduct cell culture for rubella virus. After identification of cell culture with RT-PCR, nucleotide of gene E1 of rubella virus was amplified and sequenced, followed by molecular epidemiological analysis. A total of 31 rubella viruses were isolated from 60 throat swabs. Compared 27 isolates with the WHO reference strains of all genotypes, phylogenetic tree was constructed based on the amplified 739 nucleotide fragment. These isolates belonged to two different genotypes respectively. Isolates 11009, 11052 and 11106 in 2011 belonged to genotype 2B, and others belonged to genotype 1E. Most of mutations were nonsense mutation, and sequence of amino acid was highly conserved. Amino acid sequence of most isolates of genotype 1E was identical, which suggested rubella viruses from same transmission chain might be transmitted continually since 2001. Rubella virus genotype 2B was found to be popular for the first time in Shanghai in 2011. The nucleotide sequences of these genotype 2B isolates showed 99% identity compared with that of isolates recently from Vietnam, Japan and Argentina. The resources of these strains were not confirmed due to the absence of rubella virus surveillance before.
Adolescent ; Adult ; Amino Acid Sequence ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Male ; Molecular Epidemiology ; Molecular Sequence Data ; Rubella ; epidemiology ; virology ; Rubella virus ; classification ; genetics ; isolation & purification ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; Young Adult