OBJECTIVE:To optimize the extraction technology for Huangshen electuary.METHODS:In this experiment the optimum extraction technology for Huangshen electuary was investigated by the L9(34) orthogonal test with the content of Astragaloside Ⅳ as technology index.The effects of three factors including the extraction times,time length of extraction and solvent ratio were investigated.RESULTS:The best extraction technology for Huangshen electuary was as follows:the solid-liquid ratio was 1∶10 and the decoction was carried out for 3 times(2 hours/time).CONCLUSION:The optimum extraction technology for Huangshen electuary is stable and feasible.

OBJECTIVE:To explore the construction effect of rational drug use monitoring system on the improvement of clini-cal drug use in the hospital. METHODS:The data about the construction of rational drug use monitoring system were collected from our hospital,including pharmacist workstation prescription check(check before prescription/medical order charging to realize warning in advance),clinical pharmacy management system prescription comment(realize prescription/medical order comment after-wards)and adverse drug reaction/adverse drug event(ADR)reporting information platform(realize online ADR reporting)during 2014-2015. The data was analyzed to evaluate the effectiveness of system construction. RESULTS & CONCLUSIONS:There were a total of 3 417 329 prescriptions audited by the pharmacist workstation within 2 years through improving system functions,includ-ing 7 315 prescriptions were returned to doctors to be modified,the percentage of which declined from 0.39% to 0.08%. The pass rate of prescription comment pre-judgment increased from 81.2% to 90.4% in 2015 by clinical pharmacy management system. The valid case number of ADR monitoring and reporting increased from 186 cases to 267 cases,involving all department of our hospi-tal. The construction of rational drug use monitoring system in our hospital can improve the standardization of prescription/medical orders,the level of rational drug use and the case number of ADR,which has achieved the expected results.

OBJECTIVE:To establish a method for the dissolution determination of Benidipine hydrochloride tablets.METHODS:According to Chinese Pharmacopeia(2005 edition),Benidipine hydrochloride tablets were added into hydrochloride acid 0.1 mo?lL-1 at rotation speed of 50 r?min-1,then samples were obtained after 30 min.The absorbance of samples was detected at detection wavelength of 359 nm and accumulative dissolution was calculated.RESULTS:The linear range of benidipine hydrochloride was 5～30?g?mL-1(r=0.999 9)with an average recovery of 99.76%(RSD=0.47%).The accumulative dissolutions of 3 batches of samples were all above 80%.CONCLUSIONS:Established method is simple and reliable for the dissolution determina-tion of Benidipine hydrochloride tablets.

Now syndrome study becomes the important research content in our country' TCM field,normalization of the symptoms standardization of syndromes especially becomes scientific research focal point in this field,furthermore some studies exists the conditions of symptoms short of norm and standards of syndrome differentiation discordant,as well stagnation of liver-QI with deficiency of the spleen is clinical common syndrome,so this article carried out all round analysis and investigations on the modern literature of stagnation of liver-QI with deficiency of the spleen syndrome recent ten years,to aim at analyzing the symptom characteristic and standards of syndrome differentiation on stagnation of liver-QI with deficiency of the spleen syndrome,to provide the bases of standardization and normalization of stagnation of liver-QI with deficiency of the spleen syndrome.

Objective To evaluate the efficiency of using nested PCR in restriction fragment length polymorphism analysis (P1-RFLP) for genotyping Mycoplasma pneumonia (M.pneumonia) in clinical specimens.Methods Based on the gene sequence of RepMp4 and RepMp2/3 in P1 gene of reference strains M129 (type 1) and FH (type 2),two sets of inner primers were designed with a HaeⅢ restriction enzyme site (GGCC).The nested PCR was set up to detect the target DNA in clinical specimens.The amplification products were mixed and digested with Hae Ⅲ enzyme.The genotypes were analyzed by comparing with various restriction maps and the results were verified by sequencing analysis.The concentration of DNA extracted from standard and clinical strains were detected by ten-fold dilution to evaluate the sensitivity of nested PCR-P1-RFLP and P1-RFLP.M.pneumonia-positive specimens isolated from Beijing in 2012 were analyzed by the nested PCR-P1-RFLP and the results were compared with those by P1-RFLP analysis.Results The nested PCR-P1-RFLP could effectively genotype M.pneumonia in clinical specimens and the results were consistent with those by sequencing analysis.The sensitivity of new assay was 103 times higher than that of the original P1-RFLP.Of the 115 M.pneumoniae positive clinical specimens,97.4％ (112/115) were type 1 and the rest were type 2.Conclusion The nested PCR-P1-RFLP shows high efficiency for genotyping of M.pneumonia in clinical specimens.It might be useful for the surveillance of M.pneumoniae infection.

Objective:To evaluate the rapid nucleic acid amplification detection of Mycoplasma pneumoniae (MP)-DNA and MP-RNA in the diagnosis of MP infection and therapeutic values in children. Methods:Patients who were diagnosed with pneumonia were enrolled from the Department of Respiration, Children′s Hospital of Capital Institute of Pediatrics from January 2018 to December 2018.Specimens were detected using the MP and Macrolide-Resistant isolates Diagnostic Kit (PCR Fluorescence Probing, Jiangsu Mole Bioscience Co., Ltd.) and MP Diagnostic Kit (Isothermal RNA amplification, Shanghai Rendu Biotechnology Co., Ltd.).Results:Among them, 42.1%(840 cases) of the 1 994 cases were positive for MP-DNA, and the macrolide associated gene mutations were detected in 96.0% (806/840 cases) of them, while 33.9% (551 cases) of 1 624 cases were positive for MP-RNA.Seven hundred and fifty-eight specimens were simultaneously detected by adopting MP-DNA and MP-RNA, and the positive rate was 43.1% (327/758 cases) and 36.7% (278/758 cases), accordingly, which were inconsistent (Kappa=0.604) in 613 (80.9%, 613/758 cases) cases, with significant differences ( χ2=6.60, P=0.01). Part of the specimens were rechecked with the interval of 7 days: MP-RNA was negative in 70.1% (47/67 cases) specimens and MP-DNA was negative in 36.1% (22/91 cases) specimens ( χ2=33.20, P<0.01). Conclusions:The positive detection rate of MP was at a high level in 2018, in Beijing, China.The results of MP-DNA and MP-RNA are consistant.But RNA detection can help to diagnose MP in the early stage, and monitor the survival of MP and its efficiency.

Objective To compare the capabilities of culture method, polymerase chain reaction ( PCR) and serological test in identifying Mycoplasma pneumoniae infection in children with confirmed com-munity acquired pneumonia. Methods Bronchoalveolar lavage fluid and serum samples were collected from hospitalized children with community acquired pneumonia in Capital Institute of Pediatrics from March to May in 2016. Three methods, traditional culture method, PCR and serological test, were respectively used to de-tect Mycoplasma pneumoniae infection in those children. Statistical analysis was performed by using SPSS18. 0 software and chi-square test. Results Seventy-nine children with community acquired pneumonia were enrolled in this study. Eight (10. 13%) patients were diagnosed with Mycoplasma pneumoniae infec-tions by the traditional culture method with an average positive culture period of 21 days. Twenty-three (29. 11%) patients showed positive results by using PCR analysis, including the 8 patients identified by the culture method. Forty-one (51. 90%) patients were found to be positive for Mycoplasma pneumoniae infec-tions by the serological test. However, four negative samples identified by the serological test were confirmed to be positive by PCR analysis, including two positive samples confirmed by the culture method. Statistical analysis showed that the differences in positive rates detected by using the three methods were statistically significant. Conclusion It is recommended that both serological test and PCR analysis should be used in combination with clinical symptoms for a comprehensive assessment of Mycoplasma pneumonia infection in children.

AIM: In order to study the relationship between mitochondrial deficiency and Alzheimer's disease(AD), we used sodium azide, a specific inhibitor of cytochrome C oxidase(COX), to develop a cell model of mitochondrial complex IV deficiency and investigated the impairment of microtubules and microtubule-associated proteins. METHODS: Primary cultured hippocampal neurons of hewborn rats were exposed to sodium azide ,then cell viability was measured by MTT method; cell morphology, immunofluorecence-stained cellular microtubules and microtubule-associated proteins were observed by confocal microscopy. RESULTS: Primary cultured hippocampal neurons were exposed to 8-128 mmol/L sodium azide for 3-24 h, MTT absorbance decreased dose-and time-dependently. Exposed to 64 mmol/L sodium azide for 6 h, the processes of cells retracted, synapses disappeared, axons were shortened under contrast microscope. Meanwhile, microtubles were disassembled and became disorderly, the expression of microtubule-associated proteins were also reduced especially in the processes observed by confocal microscopy. CONCLUSIONS: Sodium azide inhibits the assembly and polymerization of tubulin in microtubules which may be reduced by low expression of microtubule-associated proteins in nerve cells. The damage of axons induced by microtubule collapse further blocks the intercellular signal transduction and intracellular material transportation which are important causes in cell death.

OBJECTIVE To analyze the resistance of Enterobacter cloacae AmpC ?-lactamase with/without being induced by imipenem(IMP),cefepime(FEP),ceftriaxone(CRO) and aztreonam(ATM).The ampD genes of mutant strains were sequenced. METHODS Five wild type strains and 5 hyper-inducible type strains of E.cloacae were selected and induced by the tested antibiotics in vitro.At the same time,antibiotic susceptibility tests,3-D test,isoelectric focusing(IEF) and inhibit experiment were detected to identify hyper-producing AmpC ?-lactamase.ampD Gene sequencing was preformed in part of mutant strains. RESULTS There wasn′t obvious alteration in MIC with IMP,FEP,CRO and ATM for wild type strains whether they were induced or not.But,for the hyper-inducible type,there was apparent increasing in MIC after antibiotics inducing,especially CRO and ATM,up to 10.7-128 times.The DNA sequences analysis in the mutation strains showed there existed the replacement of the single base in multiple sites,and a few sites of amino acid were altered. CONCLUSIONS Mutant sites in ampD gene sequences are identical even though antibiotics as inducer are different.

Objective To investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) genes among clinical isolates of Klebsiella pneumoniae (K.pneumoniae) in pediatrics.MethodsA total of 131 non-duplicate clinical isolates of K.pneumoniae were collected in the Affiliated Children′s Hospital of Capital Institute of Pediatrics from 2010 to 2012.PMQR genes [qnrA, qnrB, qnrS, aac(6′)-Ⅰb-cr and qepA], mutations in the quinolone resistance-determining region (QRDR) and extended spectrum β-lactamases (ESBLs) genes in those strains were analyzed by PCR.Minimum inhibitory concentrations (MIC) of different antibiotics against those K.pneumoniae strains were determined by broth microdilution method and E-test according to the guidelines issued by the Clinical and Laboratory Standards Institute (CLSI).Transferability of the PMQR genes was examined by conjugation test with the sodiumazide-resistant Escherichia coli J53.Results Among the 131 isolates, 9.92% were resistant to quinolone and 30.5% were positive for PMQR genes, including 6.87% harboring qnrB gene, 22.9% harboring qnrS gene and 4.58% harboring aac(6′)-Ⅰb-cr gene.Neither qnrA-positive nor qepA-positive strain was detected.Among these PMQR genes-positive isolates, 90% were ESBLs-producing strains and two presented mutations in gyrA and parC genes.Conjugation test showed that these PMQR genes could be transferred horizontally and the ciprofloxacin resistance increased 2 to 32 folds in transconjugants.Conclusion This study indicates that the PMQR gene-carrying rate is high in K.pneumoniae strains isolated in paediatrics in China.Most of the PMQR gene-positive strains are also ESBLs-producing strains.The PMQR genes could be transferred horizontally in bacteria.