Soil Physical and Chemical Properties， Microorganisms and Metabolites in Different Culture Environments of Gastrodia elata / 中国实验方剂学杂志

Objective:To study the soil physical and chemical properties， microorganisms， and metabolites in different culture environments of <italic>Gastrodia elata</italic>， so as to provide scientific basis for subsequent cultivation of <italic>G. elata</italic> in multiple environments. Method:The tubersphere soil of <italic>G. elata</italic> cultured in different environments was collected for analyzing the soil nutrients， microbial numbers， and metabolite differences using the agrochemical method， plate-count method， and gas chromatography-time-of-flight mass spectrometry （GC-TOF-MS）-based non-targeted metabonomic approach. Result:The analysis of soil physical and chemical properties revealed the highest soil moisture， pH， available potassium， and available phosphorus in the spinney and the highest electrical conductivity， total nitrogen， alkali-hydrolyzable nitrogen， and organic matter in the pinewood. As demonstrated by the quantitative analysis of soil microorganisms， the cultivable microorganisms were bacteria， actinomycetes， and fungi， with the bacterial population and total microbial biomass in the spinney and the number of fungi and actinomycetes in the barren slope detected to be the largest. The ratio of bacteria to fungi （B/F value） in the pinewood was the highest， while that in the barren slope was the lowest. The results of metabonomic research demonstrated that the compositions and quantities of soil metabolites in the spinney （Z group）， pinewood （S group）， and barren slope （HD group） varied. Through comparisons between S and Z groups， between HD and Z groups， as well as between HD and S groups by orthogonal partial least squares discriminant analysis （OPLS-DA）， 18， 35， and 24 differential metabolites were separately screened out， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis yielded 5， 9， and 13 metabolic pathways. There existed a significant causal relationship of the soil physical and chemical properties and microbial numbers with the metabolites. Conclusion:The soil physical and chemical properties， microbial numbers， and metabolite changes differed significantly in different culture environments of <italic>G. elata</italic>， which were sorted by the suitability in a descending order as follows： spinney > pinewood >barren slope. The soil physical and chemical properties and microbial numbers are the crucial factors driving changes in soil metabolites， suggesting that regulating the soil physical and chemical characteristics and microbial characteristics in the culture environment is an important mechanism for maintaining the <italic>G. elata</italic>-soil-microbial symbiotic system.

Risk Factors of Intrauterine Adhesion after Hysteroscopic Resection of Endometrial Polyps / 中国医学科学院学报

Objective To analyze the risk factors of intrauterine adhesion (IUA) after hysteroscopic resection of endometrial polyps. Methods Totally 359 patients undergoing hysteroscopic resection of endometrial polyps from January 2013 to December 2016 were enrolled in this study. The clinical data of IUA group and non-IUA group were compared. Univariate analysis was performed to identify the risk factors of IUA,and multivariate Logistic regression analysis was further performed to get the independent risk factors. Results Of these 359 patients,IUA occured after operation in 56 patients (15.60%). Univariate analysis showed underlying diseases (χ=7.381,P=0.004),multiple polyps (χ=3.376,P=0.040),uterus with uterine fibroids or endometrial hyperplasia (χ=6.495,P=0.009),history of curettage(χ=31.576,P=0.000),pelvic infection (χ=8.582,P=0.001),intrauterine device (χ=7.161,P=0.006),history of cesarean section (χ=5.493,P=0.014),and multigravida were (χ=16.886,P=0.000) the risk factors of IUA. Logistic regression analysis showed other diseases of uterus(χ=19.542,P=0.026),history of curettage (χ=29.614,P=0.000),pelvic infection (χ=5.627,P=0.002),intrauterine device (χ=11.342,P=0.08),history of cesarean section(χ=8.549,P=0.035),and multigravida (χ=15.493,P=0.000) were the independent risk factors of IUA after hysteroscopic resection of endometrial polyps. Conclusion Other diseases of uterus,history of curettage,pelvic infection,intrauterine device,history of cesarean section,and/or multigravida can increase the risk of IUA after hysteroscopic resection of endometrial polyps.

Heat shock protein 90-mediated inhibition of hepatitis B virus replication in hepatic cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the effect of heat shock protein 90 (HSP90) on hepatitis B virus (HBV) replication in hepatocytes and to investigate the related molecular mechanism.</p><p><b>METHODS</b>A eukaryotic plasmid expressing human HSP90 was constructed (designated as HA-HSP90). HepG2 cells were co-transfected with HA-HSP90 and the HBV replicative plasmid HBV1.3. Expression of the exogenous HSP90 was assessed by Western blotting. Expression of the HBV surface antigen (HBsAg) was determined by enzyme-linked immunosorbent assay, and HBV replicative intermediates were detected by Southern blotting. Small interfering (si)RNAs were designed against HSP90 and TBK1 and transfected into the HepG2 cells to further assess the effects of HSP90 and its underlying mechanism. HSP90-mediated effects on the expression of interleukins IL-1b and IL-6 and the interferon response gene IFIT1 were assessed by quantitating mRNA levels with real time RT-PCR.</p><p><b>RESULTS</b>The HA-HSP90 plasmid successfully expressed exogenous HSP90 protein in HepG2 cells. The exogenous HSP90 was able to inhibit HBV replication and HBsAg expression. IFIT1 expression was up-regulated after HA-HSP90 transfection, but neither IL-1b nor IL-6 were affected. The siRNA-mediated TBK1 down-regulation had no effect on the HSP90-inhibited HBV replication.</p><p><b>CONCLUSION</b>HSP90 can inhibit HBV replication and TBK1 is not involved in this process.</p>

Real-time measurement of noradrenaline release in central nervous system / 生理学报

In order to investigate the central nervous mechanism and the diseases involved in catecholamine transmitter secretion, the dynamics of catecholamine release is studied in single cell, brain slice or in vivo. Noradrenaline is an important neurotransmitter and modulator in the central nervous system (CNS) and the peripheral nervous system (PNS). In the present paper, we first compared three real-time methods used to measure noradrenaline secretion in single cells (membrane capacitance, amperometry and confocal fluorescence microscopy imaging). Compared to the electrophysiological method and fluorescence microscopy, the basic usage of the carbon fiber electrode (CFE) in neuroscience research was presented as an example. Then, we presented a primary description of ion channels, including voltage-gated Na(+), K(+) and Ca(2+) channels in locus coeruleus (LC) neurons in rat brain slices. Finally, we presented example recordings of combined patch-clamp and amperometry measurements in LC neurons, indicating Ca(2+)-dependent quantal noradrenaline release following Ca(2+) influx through Ca(2+) channels.

Antigenicity of major hydrophilic region II of hepatitis B virus surface antigen / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study whether the substitutions at the major hydrophilic region II (MHRII) of hepatitis B surface antigen (HBsAg) will impair the antigenicity of HBsAg.</p><p><b>METHODS</b>Four recombinant plasmids expressing mutant HBsAg (mtHBsAg) P1120T, C121S, K122I and T123N were constructed. HepG2 cells were transfected with the four plasmids and a plasmid expressing G145R HBsAg. The immunoreactivity of the cells expressing mtHBsAg with P1120T, C121S, K122I, T123N and G145R were detected by immunofluorescence (IF) staining and ELISA with 4 antibodies and 7 HBsAg diagnostic kits respectively.</p><p><b>RESULTS</b>mtHBsAg with P120T was recognized by mAb1 and mAb2. mtHBsAg with C121S and K122I was not recognized by any mAbs. mtHBsAg with T123N in lysates was recognized by mAb2, but not recognized in the supernatants.</p><p><b>CONCLUSION</b>Substitutions at amino acid positions 120-123 of HBsAg strongly impaired the antigenicity of HBsAg, a fact that was not appreciated previously.</p>

Expression of interferon alpha family gene of Chinese marmot in eukaryotic and prokaryotic cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the function of interferon alpha (IFNalpha) in a Chinese marmot model of hepatitis B, we expressed the Chinese marmot (Marmota himalayana) IFNalpha family gene (IFNA) in eukaryotic cells and prokaryotic cells.</p><p><b>METHODS</b>Eukaryotic and prokaryotic expression plasmids harboring Chinese marmot interferon alpha gene with different genotypes were generated using molecular cloning technology. We detected the biological activity of all expression products by viral protection assay, and analyzed their differences and species restriction of the biological activity.</p><p><b>RESULTS</b>The Chinese marmot functional genotype IFNalpha was expressed in the baby hamster kidney (BHK) cell line, and these products protected WH12/6 cells challenged by encephalomyocarditis virus (EMCV). The Chinese marmot IFN-alpha5 also expressed in E. Coli induced by IPTG, and purified fusion protein had antiviral biological activity. The biologic activity displayed differences among different subtype IFNalpha, and it had strict species restriction.</p><p><b>CONCLUSION</b>The IFNalpha family gene of the Chinese marmot can be expressed in both eukaryotic and prokaryotic cells, and the expression products show antiviral activity in a protection assay. This study provides, for the first time, evidence that IFNalpha from the Chinese marmot has an antiviral function in vitro and can be used to improve the efficacy of current therapies for HBV infection in our Chinese marmot model.</p>

Detection of mutants of the "a" determinant region of hepatitis B surface antigen S gene among Wuhan childhood patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether the presence of HBV mutant in vaccinees simply reflects the prevalence of HBV mutant in a specific geographic area or is indeed due to the immune pressure induced by vaccination.</p><p><b>METHODS</b>HBV S genes were amplified by using polymerase chain reaction (PCR) and DNA sequence analysis of the "a" determinant was performed on sera from 30 childhood patients with immunoprophylaxis and 30 patients without vaccinations.</p><p><b>RESULTS</b>Mutations of the "a" determinant were detected in 8 of the 60 patients. They were all of the adw subtype. The prevalence of amino acid substitutions as detected by direct sequencing was higher in those fully-vaccinated than of those not vaccinated. In all 8 vaccinated and also with detectable mutants, the mean age was older than the other vaccinated children.</p><p><b>CONCLUSION</b>The prevalence of mutants is related to HBV subtypes and genotypes. Universal vaccination has accelerated an accumulation of HBsAg "a" determinant mutants with amino acid changes critical for immune escape in vaccinated children who became carriers. This suggests that new vaccination strategies should be considered.</p>

Inhibitory and excitatory effects of sodium nitroprusside on supraopticnucleus neurons in vitro / 中国临床药理学与治疗学

Aim To realize the regulatory actions of nitric oxide(NO) on supraoptic nucleus (SON)neurons by observing the effects of NO donor sodium nitroprusside(SNP) on SON neurons.Methods The cell electrophysioloical properties were obtained by the intracellular recording techniques from the SON neurons in adult rat hypothalamic slices.Results In 11 silent cells, superfusion of SNP(1 mmol?L-1) for 3～5 min resulted in the depolarization response with a decrease of membrane resistance and time constant(P