Objective:To investigate overall quality status of rose bengal agar culture medium for drug control in China to provide technical support for the drug standard revision. Methods:Four quality control microorganisms, namely Aspergillus niger, Candida al-bicans, Staphylococcus aureus and Escherichia coli, were used to evaluate the quality of rose bengal agar culture medium from 7 domes-tic and foreign manufacturers. Results:Including the control medium, the pH value of all the 8 media was different. The growth rate of Aspergillus niger and Candida albicans in the seven tested media were all above 0. 7 without significant difference (P>0. 05). The selectivity factors of Staphylococcus aureus in the seven media and the control medium was more than 6. The eight media showed differ-ences in the selectivity factors of Escherichia coli. Among them, A, B, C, E, G and the control medium had no inhibitory ( selectivi-ty) , while D and F showed certain selectivity. Conclusion:The rose bengal agar media from different manufacturers for drug control show similar growth-promoting ability with different pH value and selectivity to Escherichia coli.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Microbiological data deviations ( MDD) investigation differs from out of the standard ( OOS) investigation in chemical inspection items. Investigating Out-of-Specification( OOS) Test Results for Pharmaceutical Production issued by FDA specifically ex-cludes microbiological and other biological assays. Until recently, there is not a definitive guideline for MDD investigation in the world. Some institutions and experts have explored the practice and theory of MDD investigation. Referred to the existing practical experience and theoretical achievement and based on the provisions of Chinese GMP (2010 revision) and the requirements of Chinese Pharmaco-poeia Ⅱ(2010 edition) Appendix XIX Q, the article discussed the methods for MDD laboratory investigation.

Objective To observe the clinical efficacy of thunder-fire moxibustion at Dazhui (GV14) and Shenshu (BL23) in treating mild cognitive impairment (MCI) due to ischemic cerebral stroke. Method Sixty patients with MCI due to ischemic cerebral stroke were randomized into a treatment group and a control group, 30 cases each. The control group was intervened by joint treatment plus cognitive training, while the treatment group was by thunder-fire moxibustion in addition to that given to the control group. The two groups were evaluated by the Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE) and Wechsler Memory Scale (WMS), the major symptoms were observed, and the clinical efficacies were compared between the two groups. The two groups were both treated for 8 weeks. Result The total effective rate was 80.0%in the treatment group, versus 70.0% in the control group, and the difference was statistically significant(P<0.05). Conclusion The selected acupoints can improve the cognition and activities of daily living (ADL) in MCI patients;thunder-fire moxibustion at Dazhui and Shenshu plus cognitive training can produce a better clinical efficacy than dry cognitive training. Therefore, we can combine thunder-fire moxibustion with modern rehabilitation to enhance the therapeutic efficacy in preventing and treating MCI.

Objective:To construct recombinant lentiviral vectors harboring interference RNA ( RNAi ) targetting murine TNF-αgene,so as to lay the foundation on the RNAi gene therapy.Methods: Three small interfering RNA ( siRNA) sequences targeting murine TNF-αgene ( siRNA1,siRNA2,siRNA3) and negative-control siRNA were designed and synthesized.The inhibition effects of siRNAs on TNF-α,IL-1βand IL-6 secretion of LPS-stimulated RAW264.7 macrophages were observed using real-time PCR and ELISA methods.DNA oligo was designed and synthesized according to the most effective siRNA 2 sequence.The recombinant lentiviral shuttle plasmid expressing short hairpin RNA ( shRNA) was constructed and sequenced.The lentiviral shuttle plasmids with packaging plasmids were transfected into 293T cells to produce lentiviral particles.Results: ①The TNF-αmRNA relative expression levels of siRNA1, siRNA2 and siRNA3 were 0.24±0.01,0.16±0.02,0.19±0.01 respectively,significantly lower than that of negative control (0.95± 0.02) (F=531.3,P<0.001).The inhibition rates at mRNA level were 74.26%,83.09%,79.93%,respectively comparing with negative control.No significance was observed in IL-1βor IL-6 mRNA relative expression change after TNF-αsiRNA transfection ( P>0.05).②The TNF-αprotein expression levels of siRNA1,siRNA2 and siRNA3 were (23.95±1.21),(17.27±1.46),(19.07± 1.57)ng/ml respectively,significantly lower than that of negative control (35.37±2.93)ng/ml (F=18.1,P=0.000 6<0.001).The inhibition rates of protein expression were 32.29%, 51.16%, 46.08%, respectively comparing with negative control.③The PCR product electrophoresis showed that recombinant vectors yielded 343 bp fragments,non-constructed vectors yielded 306 bp fragments.DNA sequencing partially showed insertion sequence.④Lentiviral particles were obtained by transfecting 293T cells with recombinant lentiviral shuttle plasmids and lentiviral packaging plasmids.Cells grew well during virus production with strong fluorescence expression.The titer of concentrated virus was 2×106 TU/μl.Conclusion:The lentiviral vector harboring RNAi targeting murine TNF-αgene has been successfully constructed.

Objective:To investigate the expression of survivin and MMP-9 in the eutopic endometrium of patients with endometriosis(EMs).Methods:Eighty patients with endometriosis were selected as EMs group,four of them were in clinical stage Ⅰ,25 in stage Ⅱ,44 in stage Ⅲ and 7 in stage Ⅳ,47 in the proliferative phase and 33 in the secretory phase of the menstrual cycles.Twenty patients with hysteromyoma were selected as control group,including 7 patients in the proliferative phase and 13 in the secretory phase of the menstrual cycles.The expression levels of survivin and MMP-9 were determined by immuno-histochemical method.Image-Pro Plus 6.0(IPP),the special-purpose system for quantitative measurement of medical images,was used to analyse the results quantitatively.Results:Both survivin and MMP-9 were found positive in the plasma of the endometriosis.Both survivin and MMP-9 expression levels were higher in EMs group than those in the control group(P ＜0.01).Besides,survivin was over expressed with no relation to the menstrual cycle of the endometium.Survivin expression was higher in endometriosis stage Ⅱ than that of the stage Ⅲ(P ＞ 0.05).There was no difference in survivin expression within other stages(P ＞ 0.05).While MMP-9 expression had no difference between all the endometriosis stages(P ＞ 0.05).There was a close relationship between survivin and MMP-9 expression levels in 80 eutopic endometfium of endometriosis(r=0.262,t=2.860,P ＜ 0.05).Conclusion:The over expression of survivin and MMP-9 may play an important role in the pathogenesis of endometriosis.

Objective To explore the relating factors for cognitive dysfunction in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods Totally 150 patients with acute exacerbation of COPD (AECOPD) and 150 gender and age-matched healthy subjects (controls) received the cognitive and depression function assessment.150 AECOPD cases were classified into two groups:COPD patients with or without cognitive dysfunction.Fasting blood uric acid (UA) was measured and compared between the two groups.Relating factors for cognition dysfunction were analyzed.Results The score of Montreal Cognitive Assessment (MoCA) was reduced in AECOPD patients as compared with controls [(19.01±3.58) vs.(26.58±1.42)scores,t=-3.48,P=0.032].The score of Hamilton depression (HAMD) scale was higher in AECOPD patients than in controls [(18.05 ± 4.50) vs.(9.98 ± 3.51) scores,t =4.63,P =0.028].The prevalence of cognitive impairment in the patients with AECOPD was 82 cases (54.7％).The serum UA level was lower in the AECOPD patients with cognitive impairment than in patients without cognitive impairment [(235±42) μmol/L (n=82) vs.(332±45) μmol/L (n=68),t=-6.65,P=0.003].In AECOPD patients,the cognitive function was correlated with education,length of hospital stay,serum UA level,COPD stages,disease duration and the depression level,but was not correlated with smoking and body mass index.Conclusions The cognitive impairment is highly prevalent in patients with AECOPD,which is associated with lower serum UA level,longer hospital stay and disease severity.

In this article, an HPLC method for the contents determination of amino acids in Qihong Maitong injection was reported. In detailed, OPA-Fmoc pre-column derivatization was adopted, and related HPLC methods to determine the contents of amino acids was established. Linear relationship was well constructed for 17 amino acids through the method mentioned above. Briefly speaking, the optimized method was accurate and reproducible, and suitable for the determination of amino acids in Qihong Maitong injection and corresponding quality control.

Objective:Discussion MKK4 protein expression in nasopharyngeal carcinoma and -1044 A/T polymorphism correlation.Methods:90 patients with nasopharyngeal carcinoma , MKK4 protein expression was determined by immunohistochemical staining,polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP ) to detect the gene -1044A/T sites monocytes nucleotide polymorphism.Results:MKK4 protein expression in nasopharyngeal carcinoma (-) was 24.4%(22/90),(+) was 15.6%(14/90),(++) was 34.4%(31/90),(+++) was 25.6% (23/90).Low expression (-/+) patients with a total of 36 cases,-1044AA genotype accounted for 22 cases (61.11%),AT genotype accounted for 12 cases (33.33%),TT genotype accounted for two cases (5.56%),AT+TT gene type accounted for 14 cases (38.89%).The patients with high MKK4 expression of 54 cases,of which accounted for 38 cases of AA genotype (70.37%),AT genotype accounted with 15 cases (27.78%),TT genotype accounted for one case (1.85%),AT +TT genotype accounted for 16 cases (29.63%).Low expression and high expression of T gene mutation occurs no significant ( Z=0.323 , P=0.747 ) .Conclusion: MKK4 protein expression correlated with -1044 A/T gene promoter polymorphisms was no significant correlation .