Objective To analyze the relationship of penA, ponA, porB and mtrR gene mutations with the reduced sensitivity to ceftriaxone in N.gonorrhoeae isolates from Shenzhen city.Methods A total of 296 clinical isolates of N.gonorrhoeae were collected in Shenzhen city from 2009 to 2011.The agar dilution method was used to estimate the sensitivity of these N.gonorrhoeae to ceftriaxone.Totally, 53 strains with reduced sensitivity to ceftriaxone (minimum inhibitory concentration (MIC): 0.06-0.50 μg/ml) were identified, and 53 strains with high sensitivity to ceftriaxone were randomly selected from the remaining strains and served as the control group.PCR was performed to amplify the penA, ponA,porB and mtrR genes from the 106 isolates followed DNA sequencing.Results The mosaic structure of the penicillinbinding protein 2 (PBP2) gene (penA gene) was found in only one isolate with a ceftriaxone MIC of 0.125 0 μg/ml.Amino acid sequence analysis of the remaining 105 isolates yielded 16 different amino acid patterns.The MICs of ceftriaxone were relatively high (0.062 5 μg/ml) in N.gonorrhoeae strains harboring the amino acid patterns ⅩⅢ, ⅩⅧ or ⅩⅩⅩⅧ,but relatively low (0.008 0 μg/ml) in those harboring the amino acid pattern Ⅱ.No significant differences were observed in the frequency of mtrR, porB or ponA gene mutations between N.gonorrhoeae isolates with reduced sensitivity to ceftriaxone and those with high sensitivity (all P ＞ 0.05).Conclusions The mosaic structure of PBP2 may be not the primary reason for reduced sensitivity of Neisseria gonorrhoeae to ceftriaxone in Shenzhen, while different amino acid patterns produced by various mutations in amino acid residues at positions 500-580 in the non-mosaic PBP2, together with mtrR, porB and ponA mutations, may play more important roles in the reduced sensitivity.

Two single nucleotide polymorphisms (SNP) of adiponectin gene (SNP+45 and SNP+276) were screened in Chinese population. The results showed that the distribution of SNP+45 genotypes were significantly different and G/G genotype in type 2 diabetic population appeared more frequently than that in non-diabetic population. It suggests that adiponectin gene seems to be the susceptive gene in Chinese type 2 diabetics.

Objective To investigate the prevalence, molecular mechanism and genetic characteristics of azithromycin-resistant Neisseria gonorrhoeae (N.gonorrhoeae) strains isolated in Shenzhen.Methods N.gonorrhoeae strains were collected in Shenzhen from 2011 to 2015.Agar dilution method and E-test were used to detect the minimum inhibitory concentrations (MIC) of these strains to azithromycin.All azithromycin-resistant (AZM-R) strains (MIC≥2 μg/ml) and some azithromycin-sensitive strains (MIC≤0.25 μg/ml) which were randomly selected as the control group were screened for mutations in 23S rRNA, mtrR and erm genes and genotyped by using N.gonorrhoeae multi-antigen sequence typing (NG-MAST).Results A total of 788 N.gonorrhoeae strains were collected, 148 (18.8%) of which were AZM-R strains (MIC≥1 μg/ml).Eighteen out of 21 high-level AZM-R (AZM-HLR) strains had A2143G mutations in the four copies of the 23S rRNA gene.Twelve out of 29 middle-level AZM-R (AZ-MLR) strains had missense mutations, among which C2611T mutations in the four copies of the 23S rRNA were detected in 10 strains.Incidence of G45D/Y105H mutation in AZM-HLR strains was higher than that in AZM-MLR (χ2=12.702, P=0.000) or AZ-S (χ2=4.462, P=0.035) strains according to the analysis of the promoter and coding region of mtrR gene.PCR analysis revealed that only one strain carried ermB gene (MIC=2 μg/ml).The 788 N.gonorrhoeae strains were typed into 81 sequence types (STs) by NG-MAST, most of which were represented by one strain only.STs of ST3356 and ST1866 that were identified in the AZ-R strains in the current study had been noted in a previous report of emerging AZM-R N.gonorrhoeae strains in Nanjing, Chongqing and Guangzhou.Neighbor-joining (NJ) phylogenetic tree showed that the resistant strains did not form a separate cluster.Conclusion Currently, it is not suitable to use azithromycin as a monotherapy for gonorrhea in Shenzhen.Mutations of A2059G and C2611T in 23S rRNA of N.gonorrhoeae were respectively responsible for high-level and middle-level resistance to azithromycin.Repeated emergence of ST1866 and ST3356 will help us monitor and analyze the epidemiological characteristics of N.gonorrhoeae strains resistant to azithromycin in Shenzhen.

OBJECTIVE:To provide reference for the further implementation of national essential medicine system of township hospitals. METHODS:5% township hospitals in a province were collected as sample by using random cluster sampling method, and statistics,comparison and evaluation was made by issuing questionnaires to get health resources distribution status,health ser-vice status and income and expenditure status before(in 2009 and 2010)and after(from 2011 to 2013)the implementation of na-tional essential medicine system of township hospitals. RESULTS:Totally 46 township hospitals were surveyed and totally 46 ques-tionnaires of township hospitals were recycled. Average number of beds in township hospitals increased from 15.1 beds in 2009 to 19.4 beds in 2013. Average annual outpatients increased from 11 200 in 2009 to 16 100 in 2013,and average number of hospital discharge increased from 644 in 2009 to 924 in 2013. The proportion of government financing to total income increased from 25.9% in 2009 to 47.1% in 2013,proportion of drug income to the total income decreased from 54.9% to 29.7%,drug profit rate decreased from 25.5% to 3.7%,and proportion of township hospital under deficit decreased from 17.4% to 4.3%. Average time that residents see a doctor in township health center increased from 0.47 in 2009 to 0.74 in 2013,and cost of average hospitaliza-tion and outpatient drugs decreased from 805 to 718 and 28.1 to 24.1,respectively. CONCLUSIONS:The implementation of nation-al essential medicine system has no negative effect on outpatient service,but first inhibits then promotes the inpatient services. Na-tional essential medicine system has effectively cut down the financial burden of drugs,but it has no effect on total health burden. It is difficult to realize the excessive rapid rise of health ex-pense by the single implementation of essential medicine sys-tem,and it needs comprehensive reform,collaboration and in-teraction of medicine and health to effectively relieve the prob-lem of“expensive ill”.

OBJECTIVE To analyze the phonetic acquisition and speech articulation in deaf children. METHODS The speech articulation test and consonant and vowel identification test were performed with 30 children. RESULTS The consonants /b.p.d.t.m.l/ were the easiest to learn. Of the vowels, single vowels received the highest score. CONCLUSION The results of the consonant identification test, listed from easy to difficult, were /b.p.d.t.m.l/,/h.f.s.x.n/,/c.z.zh.ch.j.q.r/. The results of the vowel test, listed in the same order, were single, duplicate and nasal.

Objective To study the changes in miRNAs expression in the exosomes of human umbilical vein endothelial cells(HUVECs) after 60 Co γ-rays expose using microRNA(miRNA) chips and bioinformatics techniques so as to provide new clues to the mechanism of radiation-induced vascular tissue injury and its bystander effects.Methods HUVECs exosomes were collected in the control and 4 Gy irradiated cells by ultra-high-speed centrifugation,and further confirmed using transmission electron microscopy (TEM) and Western blotting of exosomes biomarkers.miRNA microarray was used to analyze miRNA expression profiles of exosomes and cells.Also,real-time quantitative PCR(qRT-PCR) was used to verify differentially expressed miRNAs,and the miRDB and TargetScan were performed to predict the target genes of the differentially expressed miRNAs.Bioinformatics analysis was performed using DAVID,KEGG and other online tools.Results Compared with the control exosomes from non-irradiated HUVECs,miRNA microarray analysis revealed that 5 up-regulated,and 13 down-regulated miRNAs were identified in the exosomes from HUVECs at 0.5 h after 4 Gy-irradiation,and 16 up-regulated and 5 down-regulated miRNAs at 2 h after 4 Gy-irradiation.Moreover,38 and 85 miRNAs were differentially expressed respectively in the HUVECs at 0.5 h and 2 h after radiation.The difference was statistically significant(P<0.01).The results of bioinformatics showed that these miRNAs might exert the radiation-induced bystander effect (RIBE) by regulating MAPK signal pathways,RAS and PI3K-Akt signal pathways.Conclusion The ionizing radiation injury significantly alters the components and expression levels of exosomal miRNAs,which play important roles in regulating the signal pathways in response to radiation.

OBJECTIVE To investigate the bone development activity and differences in safety of ethanol extract (EE) and water decoction (WD) of Psoralea corylifolia L.efficiently.METHODS Zebrafish larvae were co-exposed to prednisolone 25 μmol· L-1 and different concentrations of EE and WD (0.1,1.0,10 and 100 mg crude drug· L-1),and etidronate disodium (ED) 30 mg·L-1.All these groups were incubated at 28.5℃ until 9 dpf.The medium solution was changed every other day.Zebrafish skeleton at 9 dpf was stained with alizarin red and inspected under an optical microscope,in addition,the death toll and organ toxicity of zebrafish were also observed.The mRNA expression of osteoprotegrin (OPG) and receptor activator of NF-κB ligand (RANKL) in 9 dpf zebrafish were determined with fluorescence quantitative PCR.Zebrafish embryos (1 dpf) were exposed to various concentrations of EE (10,20,30,35,40,50 and 60 mg crude drug· L-1),WD (10,50,100,125,150,175,200 and 500 mg crude drug· L-1),psoralen (12.5,25.0,50.0,100.0,200.0 and 400.0 μmol·L-1) and bakuchiol (1,5,10,25 and 50 μmol· L-1).Embryonic morphology of zebrafish (3 dpf) was inspected with an optical microscope and the death toll of embryos or larvale was counted from 2 dpf to 9 dpf and LC50was calculated.Components of EE and WD ware analyzed by HPLC method.RESULTS Both EE (0.1 mg crude drug· L-1) and WD (1.0 mg crude drug· L-1) groups could increase the staining area and optical density values of zebrafish skeleton compared with prednisolone group (P＜0.01),indicating the increase in bone mineralization;the OPG mRNA expression in both EE and WD (1.0 mg crude drug· L-1) groups increased,while the RANKL mRNA expression decreased (P＜0.01) and the ratio of OPG/RANKL improved obviously (P＜0.01).Embryos exposed to EE,WD,psoralen and bakuchiol showed swelling of the heart and yalk sac,and decrease in GOT.The LC50 of WD and psoralen was 5～8 and 5～21 times that EE and bakuchiol,respectively.The composition and relative content of EE and WD also varied considerably.CONCLUSION Bone development activity and toxicity of EE are both stronger than those of WD.The lipid soluble characteristic components of Psoralea corylifolia L.,may be critical components of toxicity.

Objective:Nuclear apoptosis-inducing factor 1(NAIF1) is a novel apoptosis gene cloned in laboratory. To analyze the binding proteins of NAIF1 by pulldown method, the fusion expression vector of truncated human nuclear apoptosis-inducing factor 1〔NAIF1(73-327)〕 was constructed, were expressed and purified the recombinant GST-NAIF1(73-327) fusion protein in E.coli.Methods:The cDNA encoding human NAIF1(73-327) was amplified by PCR and cloned into pGEX-KG vector. The GST-NAIF1(73-327) fusion protein was expressed in E.coli and purified by affinity chromatography. The purified protein was detected by SDS-PAGE, Western blot and ESI-Q-TOF-MS/MS.Results:A prokaryotic expression vector of GST-NAIF1(73-327) was constructed and the GST-NAIF1(73-327) fusion protein was expressed in E.coli at high level. SDS-PAGE analyses indicated that the purified protein was about 53 kD. Western blot and MS/MS analyses verified the recombinant fusion protein.Conclusion:An efficient method for obtaining recombinant GST-NAIF1(73-327) fusion protein had been established and it could be used for further studies on the structure and function of NAIF1.

This study aimed at efficiently screening out the toxic herbal medicines in Zhuang Gu Guan Jie pill (ZGGJ) based on the zebrafish model.Zebrafish embryos at 1 day post fertilization (dpf) were administered by ZGGJ and the water decoction (WD) and ethanol extract (EE) of 12 herbal drugs in it with various concentrations.The number of embryo deaths after the administration was recorded at 2-6 dpf and 2-8 dpf,respectively.Embryonic micro-morphology of zebrafish (3 dpf) was observed,LC50 value at 6 dpf was calculated by SPSS16.0.As a result,both the WD and EE of ZGGJ,fructus psoraleae,radix dipsaci,olibanum,myrrh,radix angelicae pubescentis and aucklandiae radix,and the EE of Caulis Spatholobo and Rhizoma Cibotii presented high organ toxicity to the juvenile zebrafish,featuring yolk sac swelling,deforming,blackening,indicating hepatotoxicity in the zebrafish.Their LC50 value at 6 dpf was from 33.2 μg·mL-1 to 392.7 μg·mL-1 (LD50 value was converted to crude drug.The same bellow.).The WD and EE of Epimedii Folium at 1,500 μg· mL-1 led to yolk sac swelling and blackening.The LC50 at 6 dpf varied at 1,100-1,350 μg·mL-1,showing a certain toxicity of Epimedii Folium.The WD of Caulis Spatholobo and Cibotii Rhizoma and both WD and EE of Rhizoma drynariae,taxilli herba and prepared rehmanniae radix didn' t change the toxicity to the organs in zebrafish.LC50 value were over 1,500 μg·mL-1,presenting favorable safety.In conclusion,it was demonstrated that the zebrafish model efficientlyidentified the toxicity of different herbal medicines from ZGGJ and efficiently screened out the potential toxicants in vivo with the advantages of real-time,dynamic and large scale for traditional Chinese medical compounds.

OBJECTIVE To explore near 9-year monitoring of hospital environmental health study to analyze the results of conventional research and take corresponding measures to the problem.METHODS Hospital Infection Control Branch Department full-time staff and part-time nurses were monitored by conventional monitoring of environmental hygiene,the results of an analysis of seasonal climate for different environmental factors and air-conditioning.Corresponding measures,with particular emphasis on air conditioning filters clean and disinfect through the monitoring results of 4000 cases for the control group and experimental group were compared at the same period a year.RESULTS Take appropriate measures before and after the monitoring results of environmental health science and the infection rate appeared too high a failure rate of 3-10 in the month of the month especially,the use of the control group occurred in a failure rate of 1.5%,Test group to adopt corresponding measures after the failure rate of 0.7%,2000-2008 can be seen from Table 1 the incidence of hospital infection in three hours before the measures taken before and after the distribution of differences(P