Abstract: Our previous work revealed berberine can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, which suggested that berberine has synergistic antifungal activity with fluconazole. Preliminary SAR of berberine needs to be studied for the possibility of investigating its target and SAR, improving its drug-likeness, and exploring new scaffold. In this work, 13-substitutited benzyl berberine derivatives and N-benzyl isoquinoline analogues were synthesized and characterized by 1H NMR and MS. Their synergetic activity with fluconazole against fluconazole-resistant Candida albicans was evaluated in vitro. The 13-substitutited benzyl berberine derivatives 1a-1e exhibited comparable activity to berberine, which suggested that the introduction of functional groups to C-13 can maintain its activity. The N-benzyl isoquinolines, which were designed as analogues of berberine with its D ring opened, exhibited lower activity than berberine. However, compound 2b, 2c, and 4b showed moderate activity, which indicated that berberine may be deconstructed to new scaffold with synergistic antifungal activity with fluconazole. The results of our research may be helpful to the SAR studies on its other biological activities.
Antifungal Agents ; pharmacology ; Berberine ; pharmacology ; Candida albicans ; drug effects ; Drug Resistance, Fungal ; Drug Synergism ; Fluconazole ; pharmacology ; Isoquinolines ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo analyze the epidemiological characteristics of hand foot and mouth disease (HFMD) in Ningbo.

METHODSA descriptive analysis was conducted through the surveillance data of HFMD in Ningbo, Zhejiang province, from 2008 to 2011. Genes on EV71 and Cox A16 were amplified with RT-PCT from the stool samples of HFMD patients. Sequences were analyzed by bioinformatics software.

RESULTS37 524 cases of HFMD were reported from 2008 to 2011, including 196 severe cases and 12 deaths. The reported incidence was 145.26 per 100 000 and the case fatality was 0.03%. Cases in children aged 5 or younger accounted for 95.89%, and the scattered cases accounted for 64.10%. Xiangshan and Ninghai counties had the highest incidence rates in Ningbo. The peak of incidence was from April to July. The number of male patients was obviously higher than females. 2394 cases of HFMD were laboratory confirmed and EV71 with the predominant epidemic strain. Data from phylogenetic analysis revealed that EV71 isolated from HFMD patients in Ningbo belonged to C4a evolution branch of C4 sub-genotype, with several transmission chains. Cox A16 belonged to B1 evolution branch. 53.48% of the healthy children in Ningbo showed EV71 antibody positive. The geometric mean of the antibody titer (GMT) was 11.23 (8.33 - 14.98) in healthy children. Cox A16 antibody was detected at 63.18% of the healthy children in Ningbo. GMT in healthy children was 12.61 (6.70 - 16.52).

CONCLUSIONHFMD was highly endemic in Ningbo, with children under 5 years old were at high-risk. The major etiologic agent was EV71 which belonged to C4a in the C4 sub-genotypes. Cox A16 belonged to the B1 evolution branch, which were in line with the predominant virus circulating in the mainland of China.

Child ; Child, Preschool ; China ; epidemiology ; Coxsackievirus Infections ; epidemiology ; Enterovirus ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Incidence ; Infant ; Infant, Newborn ; Male

Objective To know the immunologic response at different periods with different hepatitis B vaccine(HepB)in infants.Methods From July 1 to December 31 in 2009,the local infants born from hepatitis B virus(HBV)surface antigen (HBsAg)negative mother in Ninghai were selected and received a 0.5 ml of 5 -μg HepB made by recombinant deoxyribonucleic acid (DNA)techniques in saccharomyces cerevisiae yeast (HepB -SCY)and those in Yuyao were selected and received a 0.5 ml of 10 -μg HepB made by recombinant DNA techniques in hansenula polymorpha yeast (HepB -HPY)with a 0 -1 -6 month schedule.The specimens of blood were collected at 7 months,12 months and 18 months after vaccination for testing antibody.Results At 7 months,12 months and 18 months after vaccination,the geometric mean concentration (GMC)of Anti -HBs in HepB -SCY group were 714.79 ±3.50 mIU /mL,259.89 ± 3.87 mIU /mL and 78.38 ±4.04 mIU /mL,respectively.While in HepB -HPY group were 2 420.76 ±3.51 mIU /mL, 703.55 ±3.46 mIU /mL and 171.32 ±3.76 mIU /mL,respectively.There was statistical significant difference of GMC between HepB -SCY group and HepB -HPY group(P <0.01).The percent of antibody <100 mIU /mL in HepB -SCY group were 6.39%,23.33% and 55.42%,and in HepB -HPY group,were 2.17%,5.17% and 31.95%,respectively. The difference between HepB -SCY group and HepB -HPY group had significantly statistical significance (P <0.01 ). Conclusion The immunologic response at different periods of HepB -HPY was better than that of HepB -SCY.

OBJECTIVETo study the potential risk factors of human infecting with Streptococcus suis.

METHODS1: M matched case-control study was conducted. 29 human cases of Streptococcus suis infection in the early phase were included in the case group, Patients' family members, neighbors and peoples who had worked together with patients to handle deceased or sick pigs in the last week were recruited as matched controls. There were 147 controls in total. Both cases and controls received questionnaire investigation including the ways to contact sick/dead pigs. Conditional logistic regression was employed to analyze matching data.

RESULTSAccording to the results of multivariate analysis, slaughtering (OR = 11.978, 95% CI: 3.355-42.756), carcasses cutting and processing (OR = 3.008, 95% CI: 1.022-8.849) sick/dead pigs were associated with cases related to human Streptococcus suis infection. The attributable risk proportion were 91.65% and 66.76% respectively. The other types of exposures to sick/ dead pigs, including feeding, selling, burying and eating, were not associated with the human Streptococcus suis infection in our study population.

CONCLUSIONSlaughtering, carcasses cutting and processing sick/dead pigs were important risky behavior for humans to be infected by Streptococcus suis.

Adult ; Aged ; Case-Control Studies ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Occupational Exposure ; adverse effects ; statistics & numerical data ; Risk Factors ; Streptococcal Infections ; epidemiology ; etiology ; microbiology ; Streptococcus suis ; physiology

Objective: To identify the status and determinants of sharing personal HIV information with sexual partners among men who have sex with men (MSM) meeting their casual sexual partners on the internet. Methods: A cross-sectional study was conducted in five cities (Hangzhou, Ningbo, Wenzhou, Taizhou and Shaoxing) in Zhejiang province. The recruitment was enrolled by MSM social organization and in voluntary counseling and testing clinics, with a sample size of 793. A self-designed network questionnaire collected essential characteristics, HIV knowledge, sexual behavior, and sharing personal HIV status. SPSS 20.0 software was used for statistical analysis. Results: Among 767 MSM enrolled 302 MSM who reported finding casual sexual partners on the internet were enrolled in the analysis. MSM reported finding casual partners on the internet only, finding sexual partners online and in places were 62.6% (189/302) and 37.4% (113/302), respectively. Among those reporting web-based sexual behavior in the last six months, 54.6% (165/302) informed their partners of their HIV status, 49.2% (146/297) inquired about HIV status, and 42.9% (82/191) knew HIV status before sex intercourse, 75.8% (113/149) reported consistent condom use with HIV negative partners. The multivariable logistic regression analysis showed that related factors of inconsistent inquired HIV status of partners included 25-34 years old (aOR=2.17, 95%CI: 1.20-3.91), >2 partners on the internet in the last six months (aOR=2.13, 95%CI: 1.27-3.57), low-risk perception of HIV infection with online partners (aOR=1.96, 95%CI:1.14-3.35), numbers of HIV testing >1 times (aOR=0.38, 95%CI: 0.22-0.66). Conclusions: The willingness to know the HIV status of partners among MSM who met sexual partners on the internet was high but with a low rate of knowing their sex partner's HIV status in Zhejiang province. However, the successful implementation proportion was low. Therefore, it is necessary to pay attention to people who are elderly, with less conscience about the risk of the sex partners on the internet, have more sex partners, and have received few HIV tests. In addition, peer education was needed to promote related intervention programs.
Aged ; Male ; Humans ; Adult ; Sexual Partners ; Homosexuality, Male ; HIV Infections ; Cross-Sectional Studies ; Sexual and Gender Minorities ; Sexual Behavior ; Information Dissemination ; Internet

A high performance liquid chromatography charged aerosol detector (HPLC-CAD) method was established for the simultaneous determination of five saponins (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, Ginsenoside Rd) in Yaobitong capsule, providing a method for quality control. The sample was extracted with methanol and chromatographic separation was performed on a Waters Xbridge Phenol column (150 mm×4.6 mm, 3.5 μm) using acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃ and the injection volume was 10 μL. The nebulizer temperature of CAD was 35 ℃ and the air pressure was 60.2 psi, the filtration was 3.6 s, and the collection frequency was 5 Hz. Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd showed a good linear relationship in the range of 16.96-203.5 μg·mL^-1 (R² = 0.999 3), 54.46-653.5 μg·mL^-1 (R² = 0.999 3), 10.96-131.5 μg·mL^-1 (R² = 0.999 6), 51.50-618.0 μg·mL^-1 (R² = 0.999 0), 15.94-191.3 μg·mL^-1 (R² = 0.999 4), respectively. The average recoveries were 98.96%, 100.8%, 94.76%, 100.1%, 103.1%, and RSDs were 0.87%, 1.46%, 1.85%, 2.06%, 0.96% (n = 6), respectively. The proposed method is accurate, simple and reliable, and can be used for the determination of five saponins in Yaobitong capsule.

OBJECTIVEIn mid-July 2005, five patients presented with septic shock to a hospital in Ziyang city in Sichuan, China, to identify the etiology of the unknown reason disease, an epidemiological, clinical, and laboratory study were conducted.

METHODSAn enhanced surveillance program were established in Sichuan, the following activities were introduced: active case finding in Sichuan of (a) laboratory diagnosed Streptococcus suis infection and (b) clinically diagnosed probable cases with exposure history; supplemented by (c) monitoring reports on meningococcal meningitis. Streptococcus suis serotype 2 infection was confirmed by culture and biochemical reactions, followed by sequencing for specific genes for serotype and virulence factors.

RESULTSFrom June 10 to August 21, 2005, 68 laboratory confirmed cases of human Streptococcus suis infections were reported. All were villagers who gave a history of direct exposure to deceased or sick pigs in their backyards where slaughtering was performed. Twenty six (38%) presented with toxic shock syndrome of which 15 (58%) died. Other presentations were septicaemia or meningitis. All isolates were tested positive for genes for tuf, species-specific 16S rRNA, cps2J, mrp, ef and sly. There were 136 clinically diagnosed probable cases with similar exposure history but incomplete laboratory investigations.

CONCLUSIONAn outbreak of human Streptococcus suis serotype 2 infections occurred in villagers after direct exposure to deceased or sick pigs in Sichuan. Prohibition of slaughtering in backyards brought the outbreak to a halt. A virulent strain of the bacteria is speculated to be in circulation, and is responsible for the unusual presentation of toxic shock syndrome with high case fatality.

Animals ; Bacteremia ; epidemiology ; microbiology ; China ; epidemiology ; Disease Outbreaks ; Humans ; Meningitis, Bacterial ; epidemiology ; microbiology ; Shock, Septic ; epidemiology ; microbiology ; Streptococcal Infections ; epidemiology ; microbiology ; veterinary ; Streptococcus suis ; isolation & purification ; Swine ; Swine Diseases ; microbiology

ObjectiveTo predict the potential targets and mechanism of Jingfang mixture in the treatment of H1N1 influenza and provide references for clinical application of Jingfang mixture. MethodThe active components and targets of Jingfang mixture against H1N1 influenza were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP），SwissTargetPrediction， and TargetNet. The targets of H1N1 influenza were obtained from GeneCards，Online Mendelian Inheritance in Man （OMIM）， and DisGeNET and standardized by UniProt KB. The intersection targets were obtained by Venny 2.1.0. The "drug-component-target" network was constructed with Cytoscape 3.2.1 and analyzed for the topological attributes. The intersection targets were uploaded to STRING 11.5 to obtain the protein-protein interaction （PPI） network. Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis were carried out by Metascape. Finally，the top active components ranked by degree were docked to the core targets by Autodock vina and visually analyzed by PyMOL. Balb/c female rats were used for experimental verification. Hematoxylin-eosin（HE） staining was used to observe the pathological changes in lung tissues. Enzyme-linked immunosorbent assay（ELISA）was used to detect the levels of tumor necrosis factor-α（TNF-α），interleukin-10（IL-10）， and interleukin-17（IL-17）. Real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels in lung tissues. ResultThere were 144 active components in Jingfang mixture. A total of 421 target genes of Jingfang mixture and 2 956 targets of H1N1 influenza were identified，including 199 common targets. Topological analysis showed that the core components of Jingfang mixture against H1N1 influenza included quercetin，luteolin， and kaempferol，and the core targets included prostaglandin-endoperoxide synthase 2（PTGS2），estrogen receptor alpha（ESR1），inducible nitric oxide synthase 2（iNOS2），peroxisome proliferator-activated receptorγ（PPARγ），and cyclooxygenase-1（PTGS1）. GO enrichment yielded 697 items in biological process （BP） （P<0.01）， 59 items in molecular function （MF）（P<0.01）， and 21 items in cellular component （CC） （P<0.01）. A total of 132 signaling pathways （P<0.01） were obtained by KEGG enrichment analysis， including phosphatidylinositol 3-kinases（PI3K）/protein kinase B（Akt） signaling pathway and mitogen-activated protein kinase（MAPK） signaling pathway，most of which were related to the regulation of immune inflammation. Molecular docking showed that the binding energy of the active components of Jingfang mixture to the core targets was less than -5.0 kcal·mol^-1，indicating good binding activity. HE staining showed that the lung tissues were significantly improved after drug intervention，and Real-time PCR and Western blot showed that Jingfang mixture could reduce the mRNA and protein expression of PI3K and Akt in lung tissues. ConclusionJingfang mixture can play an anti-viral effect against the influenza A virus through multiple components，multiple targets， and multiple pathways. The active components quercetin，luteolin， and kaempferol may control the inflammation and regulate immunity on the PI3K/Akt，MAPK， and other signaling pathways by acting on targets such as PTGS2，ESR1，iNOS2，PPARγ， and PTGS1.

Rosmarinic acid is one of anti-tumor ingredients in the Sarcandra glabra. After treatment with 0, 10, 30, 90, 270 and 810 μmol·L⁻¹ rosmarinic acid for 24, 48, 72 hours respectively, the inhibitory effects on MCF-7 and MDA-MB-231 cells were observed by CCK-8 and cell wound healing test. No significant inhibition effecton proliferation and migration was observed in MCF-7 cell. However, 90, 270 and 810 μmol·L⁻¹ rosmarinic acid could inhibit the proliferation and migration of MDA-MB-231 cells in a dose-dependent and time-dependent manner. Flow cytometry was further used to detect apoptosis ratios of MDA-MB-231 cells after Annexin V-FITC/PI staining, and significant apoptosis was observed after rosmarinic acid treatment. Real-time PCR and Western blot tests were carried out to detect the expressions of apoptosis-related genes. The down-regulation of the Bcl-2 expression and the up-regulation of the Bax expression were observed in MDA-MB-231 cells after rosmarinic acid treatment. The results suggested that rosmarinicacid can inhibit the proliferation and migration, and induce the apoptosis of MDA-MB-231 cells, which may be correlated with the down-regulation of Bcl-2 gene and the up-regulation of Bax gene.

Near-infrared spectroscopy(NIRS) combined with band screening method and modeling algorithm can be used to achieve the rapid and non-destructive detection of the traditional Chinese medicine(TCM) production process. This paper focused on the ginkgo leaf macroporous resin purification process, which is the key technology of Yinshen Tongluo Capsules, in order to achieve the rapid determination of quercetin, kaempferol and isorhamnetin in effluent. The abnormal spectrum was eliminated by Mahalanobis distance algorithm, and the data set was divided by the sample set partitioning method based on joint X-Y distances(SPXY). The key information bands were selected by synergy interval partial least squares(siPLS); based on that, competitive adaptive reweighted sampling(CARS), successive projections algorithm(SPA) and Monte Carlo uninformative variable(MC-UVE) were used to select wavelengths to obtain less but more critical variable data. With selected key variables as input, the quantitative analysis model was established by genetic algorithm joint extreme learning machine(GA-ELM) algorithm. The performance of the model was compared with that of partial least squares regression(PLSR). The results showed that the combination with siPLS-CARS-GA-ELM could achieve the optimal model performance with the minimum number of variables. The calibration set correlation coefficient R_c and the validation set correlation coefficient R_p of quercetin, kaempferol and isorhamnetin were all above 0.98. The root mean square error of calibration(RMSEC), the root mean square error of prediction(RMSEP) and the relative standard errors of prediction(RSEP) were 0.030 0, 0.029 2 and 8.88%, 0.041 4, 0.034 8 and 8.46%, 0.029 3, 0.027 1 and 10.10%, respectively. Compared with the PLSR me-thod, the performance of the GA-ELM model was greatly improved, which proved that NIRS combined with GA-ELM method has a great potential for rapid determination of effective components of TCM.
Algorithms ; Ginkgo biloba ; Least-Squares Analysis ; Plant Leaves ; Spectroscopy, Near-Infrared