Network pharmacology-based study on mechanisms of antipyretic action of Pueraria radix / 药学学报

This study was designed to explore the antipyretic mechanism of Pueraria radix. The method of network pharmacology was used to determine the known ingredients corresponding to Pueraria radix, predict the drug-related gene /protein targets, and analyze the interplay between key ingredients and targets. Biological Information Annotation Databases (DAVID) was used to enrich the biological processes and pathways. The result of network analysis was validated by molecular docking. It was found that 49 active ingredients of Pueraria radix not only regulate 21 targets (e.g. PTGS2, EGFR), but also affect 11 biological processes (e.g. oxidation-reduction process, prostaglandin synthesis, positive regulation of fever generation and inflammatory response) and 7 metabolic pathways (arachidonic acid metabolism, serotonergic synapse and HIF-1, et al). Molecular docking results showed that more than 65% of the active ingredients could be well docked with key targets, and the relevant literatures indicated that the active components could inhibit the expression of PTGS2, which means the result has a high reliability. These results indicated that Pueraria radix may carry its pyretic action via a "multi-ingredients-multi-targets-multi-pathways" mode, which provides a scientific basis for further research and drug development.

Pharmacokinetic study on combined application of gastrodin and puerarin in rats / 中国中药杂志

To establish a HPLC method for simultaneously determining plasma concentrations of gastrodin (Gas) and its metabolites hydroxybenzyl alcohol (HBA), puerarin (Pur) and internal standard (IS) p-hydroxyphenylethanol (Tyr) in rats and studying the pharmacokinetic process and interactions of gastrodin and puerarin after single and combined intravenous injection and oral administration. With Tyr as the internal standard, plasma samples were processed with methanol for protein precipitation, supernatant was dried with N2, and residues were re-dissolved with acetonitrile-0.05% phosphoric acid (20: 80). Chromatography was carried out on an Agilent ZORBAX SB-Aq C18 column (4.6 mm x 250 mm, 5 μm), with acetonitrile-0.05% phosphoric acid as the gradient mobile phase for the gradient elution. The UV detector wavelength was set at 221 nm for Gas HBA and IS and 250 nm for Pur. After the single or combined administration of Gas and Pur, their plasma concentrations in rats were detected. WinNonlin 5.2 pharmacokinetic software and SPSS 17. 0 software were used to respectively calculate pharmacokinetic parameters of each group, make a statistical analysis and compare the pharmacokinetic processes of Gas and Pur after the single or combined administration. According to the results, the absolute recoveries between low, media and high concentrations of Gas, HBA and Pur and IS as well as Tyr were more than 77.20%, with a good linearity (r > 0.999 6, n = 5) for Gas, HBA and Pur within concentration ranges of 0.10-101, 0.03-7.58 and 0.05-5.98 mg xL ('1) respectively. The lower limits of quantification for Gas, HBA and Pur were 0.10, 0.03, 0.05 mg x L(-1), respectively. Their in-ra-day and inter-day precisions were less than 12% with the accuracy between 85. 1% -1 10. %. All of the three substances and IS were stable during the whole analysis process. The findings showed significant differences in the main in vivo pharmacokinetic parame-ers in rats (AUC, C.(max) T,½ T.(max) MRT) after the single and combined administration of Gas and Pur. Either after the oral adminis-ration or after the intravenous injection, parameters showed a lower clearance rate ( L) longer mean residence time ( RT) and higher relative bioavailability, especially after the oral administration. Specifically, the relative bioavailability of the combined oral ad-inistration of Pur was 10. 7 times of that of the single administration, while that of Gas was 1. times of that of the single administra-ion. The combined administration of Gas and Pur can promote the absorption, decrease the elimination rate and prolong the mean resi-ence time. The method is simple and accurate and can be applied in the simultaneous determination of plasma concentrations of Gas, HBA and Pur in rats and the pharmacokinetic studies.

Polymerase region mutations and hepatitis B virus genotypes in chronic hepatitis B patients with poor response to lamivudine / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the mutations in Polymerase region and hepatitis B virus (HBV) genotypes in chronic hepatitis B patients with poor response to Lamivudine treatment.</p><p><b>METHODS</b>631 chronic hepatitis B patients with poor response to Lamivudine were recruited in this study. Real-time PCR and DNA sequencing were used to determine HBV genotypes; direct sequencing was performed to detect mutations, and real-time PCR was used to quantify HBV DNA load. Mutations in polymerase region were investigated in different HBV genotypes.</p><p><b>RESULTS</b>272 patients were infected with HBV of genotype B, and 359 patients were infected with HBV of genotype C. The mean age of patients infected with HBV of genotype C (39.1+/-11.4 years old) were significant higher than that of patients infected with HBV of genotype B (33.7+/-9.7 years old) (t = -6.55, P less than 0.01). The patients infected with HBV of genotype C had relatively higher HBV DNA load [(5.96+/-1.22) log10 copies/ml] than the patients infected with HBV of genotype B [(5.58+/-1.21) log10 copies/ml] (t = -2.01, P less than 0.05). The overall incidence rate of A181V/T mutation in genotype C (5.3%) was significantly higher than that in genotype B (0.4%) (x2=12.23, P less than 0.01), but the incidence rate of M204I/V, L180M, T184A/G/I/S, S202G/I and V173L mutations was not significantly different between genotype B and C (each P more than 0.05). M204I mutation in genotype B (20.6%) was more frequent than that in genotype C (13.9%) (x2=4.91, P less than 0.05). The Lamivudine resistance mutations were not significantly different between genotype B and genotype C (x2 = 0.00, P more than 0.05).</p><p><b>CONCLUSIONS</b>The incidence rate of lamivudine resistance mutation is not significantly different between genotype B and genotype C, but patients infected with HBV of genotype C have higher HBV DNA load than patients infected with HBV of genotype B.</p>

Viral load test conducive to excluding negative subjects from suspects in HIV antibody detections / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study whether plasma viral load testing is helpful to exclude ones free from Human immunodeficiency virus (HIV) infections from suspects in HIV antibody detections.</p><p><b>METHODS</b>19 Specimens, which showed disconcordant results of the two HIV EIA testing (S/CO < 6) and indeterminated results of Western blot (WB) test, were selected. Viral load of the specimens were detected. A six-month follow up survey in detecting HIV antibody was conducted in these subjects.</p><p><b>RESULTS</b>None of these 19 cases was observed to be positive HIV viral loads and there was no any progress in WB bands development during the follow-up period. The possibility of HIV infection could be excluded.</p><p><b>CONCLUSION</b>When the specimens react with very low intensity in both EIA and WB, negative viral load result is conducive to exclude negative subjects from suspects in HIV antibody detections.</p>

Epidemiological study on the status of HIV/STDs and relative behaviors among MSM in Beijing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the infected status of human immunodeficiency virus (HIV), sexually transmitted disease (STD) and relative risk behaviors of men who have sex with men (MSM) in Beijing.</p><p><b>METHODS</b>Respondent-driven sampling (RDS) was adopted to recruit target population and to conduct behavior and serological studies among MSM. RDSAT 5.5 was used to analyze data.</p><p><b>RESULTS</b>Among 427 MSM, the age distribution was as follows: below 30 and less than 35 were 65.4% and 81.0% respectively. 69.2% (95% CI: 63.9-75.5) of them had 12 years or less of education. 73.3% (95% CI: 66.7-79.8) of them were non-Beijing registered residents. The urban and rural registered residents almost accounted for half of all the recruits. HIV positive rate was 4.6% (95% CI: 2.2-7.6) while 56.3% (95% CI: 50.9-62.5) of them reported having had bisexual sex preference. Only 14.8% (95% CI: 10.8-19.6) of them had ever had HIV test, while 22.8% (95% CI: 18.7-27.8) reported ever having had STDs. 55.3% (95% CI: 49.3-61.9) and 55.1% (95% CI: 48.7-61.3) of them had unprotected insert and receptive anal sex over the last 6 months.</p><p><b>CONCLUSION</b>Data from the HIV positive rate showed that there was a trend of increase among MSM in Beijing which called for urgent care to them.</p>

Subtype and sequence analysis of gag and env genes among HIV-1 strains circulating in Beijing residents during 2006 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the subtype distribution and the prevalence of sequence characteristics of HIV-1 strains in Beijing residents during 2006 and to analyze the relationship between distribution of HIV-1 subtypes and transmission routines.</p><p><b>METHODS</b>Blood samples from 32 new confirmed HIV-1 infected individuals from Beijing residents in 2006 and separated plasma specimens were collected. RNAs were extracted and the gag and env gene were amplified by RT-PCR and nest-PCR. PCR products were sequenced directly and phylogenetic analyses of gag and env gene were performed using the MEGA2 software.</p><p><b>RESULTS</b>Among 32 HIV-1 plasma samples, 22 gag and 4 env gene fragments were amplified and analyzed. Five HIV-1 subtypes or circulating recombinant forms(CRFs) of HIV-1 including Thai B (2 strains), B (9 strains), C (2 strains), CRF07_BC (5 strains), CRF01 AE (4 strains) were identified being circulated in Beijing. The gene divergences of gag gene inside the subtypes were 6.6%, 4.3%, 6.8%, 4.9% and 3.0% in subtype B, Thai B, C, CRF01_AE and CRF07_BC respectively. Subtypes B were predominant in Beijing, accounted for 40.9% among 22 samples.</p><p><b>CONCLUSION</b>Five HIV-1 subtypes were identified in Beijing and the surveillance of HIV-1 gene variation should be paid more attention to.</p>

Field evaluation of alternative testing strategies for the detection of HIV infection in Beijing / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify a cost-efficient alternative antibody testing strategy for screening and confirmation of HIV infection by rapid simple tests (RSTs) and enzyme-linked immunosorbent assays (ELISAs).</p><p><b>METHODS</b>Four RSTs (RST1, RST2, RST3, and RST4) and five ELISAs (ELISA1, ELISA2, ELISA3, ELISA4, and ELISA5) were evaluated in two phases by using banked and serum specimens prospectively collected at regional hospitals and voluntary counseling and testing (VCT) centers in Beijing. A total of 200 banked serum specimens were included in the first phase, including 62 HIV-positive, 127 HIV-negative and 11 indeterminate specimens. All specimens were tested by four RSTs and five ELISAs respectively. The second phase involved prospective testing of 389 routine specimens, including 92 HIV-positive, 287 HIV-negative, and 10 indeterminate specimens. All the specimens were tested by two RSTs (RST2 and RST4) and three ELISAs (ELISA1, ELISA3, and ELISA4), which were selected for their respective excellent sensitivity and/or specificity. Western blot (WB) was used as a gold standard for confirming the reactivity of all the specimens.</p><p><b>RESULTS</b>Sensitivity, specificity, and efficacy were calculated for each assay in two phases. In the first phase, four assays (ELISA4, RST2, RST3, and RST4) had a specificity of 100%. For the determination of efficacy, ELISA4, RST2, and RST4 were selected in the second phase. ELISA1 and ELISA3 which have a sensitivity of 95.9% and 93.2% respectively also entered this phase. In the second phase, all the five assays (ELISA1, ELISA3, ELISA4, RST2, and RST4) had a sensitivity and specifity of over 90%. ELISA1 had a sensitivity of 99% and ELISA4 a specificity of 99%.</p><p><b>CONCLUSION</b>The sensitivity ELISA1 and the specificit of ELISA4 are comparable to ELISA/WB standard strategy. Application of this alternative testing strategy provides a cost-effective method for determining HIV prevalence in Beijing.</p>

IgM to recombinant antigen E2 of HEV envelope protein is a sensitive and specific serological marker in diagnosis of acute hepatitis E / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To evaluate the sensitivity and specificity of IgM to recombinant antigen E2 of HEV envelope protein in the diagnosis of acute sporadic hepatitis E.</p><p><b>METHODS</b>anti-E2 IgM was detected in sera samples from 176 cases of acute sporadic hepatitis E and 191 cases of acute non A-E hepatitis by ELISA and was compared with HEV IgM detected by some domestic and Genelabs (GL) kits. HEV RNA was also detected in sera positive for anti-E2 IgM. Logistic regression was used to analyze factors associated with the detection of anti-E2 IgM and HEV RNA.</p><p><b>RESULTS</b>Anti-E2 IgM was found in 68.75% of the serum samples from the 176 patients of acute hepatitis E and the positive rate of HEV IgM detection by domestic kits was 56.25% (chi2 IgM = 6.49, P < 0.05). There were 37 (19.37%) anti-E2 IgM positive cases in those 191 sera of the acute non A-E hepatitis, out of which 11 cases were also detected as positive by the GL kit. Of the 158 anti-E2 IgM positive sera, HEV RNA was found in 81 (51.27%), among which 57.02% was positive in acute hepatitis E and 32.43% in acute non A-E hepatitis. No HEV RNA was found in the anti-E2 IgM negative sera from the cases of acute hepatitis E. By logistic regression analysis, no variance relative to the detection of anti-E2 IgM was found with the time interval from onset to hospitalization, age, levels of bilirubin, ALT and AST of the serum. Only the level of serum ALT was found being significantly associated with the detection of HEV RNA (P = 0.024).</p><p><b>CONCLUSIONS</b>(1) anti-E2 IgM is a sensitive and specific serological maker for diagnosing an acute infection of HEV. (2) HEV is still the pathogen of some cases diagnosed clinically as non-A-E hepatitis. (3) Persistent HEV viremia is possibly an important factor influencing the severity of acute hepatitis E.</p>