1.Genotype of human papilloma virus among 1 074 male outpatients
Kedi WANG ; Dongjiang XU ; Jianrong SU
Chinese Journal of Laboratory Medicine 2016;39(8):625-628
Objective To understand the prevalence and genotype distribution of human papillomavirus among male patients attending to venereal outpatient department and provide basic data for prevention and treatment of HPV.Methods Retrospective analysis method was used to analyze 1 074 patients from venereal outpatient department of Beijing Friendship Hospital during January to August in 2015.Swab specimen were analyzed by flow-through hybridization and gene chip to detect the type of HPV.Chi-square test was used to compare the distribution of CA and suspected patients.Results Among the 434 CA samples, the positive rate was 72.6%(315/434).The 58.1%(252/434) samples were high risk HPV positive and 14.5%(63/434) samples were low risk HPV positive.In high risk HPV infection, multiple infection accounted for 40.4%(175/434) and single infection was 17.7%(77/434), while in low risk HPV infection, single infection accounted for 12.9%(56/434).HPV-11, HPV-6, HPV-16, HPV-52, HPV-58 and HPV-51 were common.The positive rate among suspected CA patients was 36.6%(234/640) , and dominated in high risk HPV infection 25.3%( 162/640 ) .The positive rates of high risk HPV in CA patients [ 40.4%( 175/434 ) and 17.7%( 77/434 ) ] were obviously higher than that of suspected ones [12.9%(56/434) and 1.6%(7/434)], χ2 =95.956 and 9.122, both P<0.05.Conclusions Male patients from venereal outpatient department have a high prevalence of HPV, and common genotype are HPV-11, HPV-6, HPV-16, HPV-52, HPV-58 and HPV-51.The intensity of HPV screening should be strengthened in order to provide the vital basis for the prevention and treatment HPV related diseases.
2.Correction of the Impact of Chylemia on Hematology Analysis by Plasma Exchange and Formula Calibration Method
Kedi WANG ; Dongjiang XU ; Jianrong SU
Journal of Modern Laboratory Medicine 2017;32(3):137-139,143
Objective Comparison the corrective effect of plasma exchange method and formula method for chylemia on hematology analysis.Methods A total of 10 samples without hemolysis,jaundice and lipemia were set as control group,each sample was divided into 4 parts,then 5,10,20 and 40 μl lipid emulsion was added.Each chylous samples was treated by plasma exchange method for two times,and routine blood test was reanalyzed with hematology analyzer.The comparisons before and after the exchange was made,while the test results on each exchange were analyzed.For another part,the HGB of chylous plasma was tested,the value was substituted into HGBcorrected value =HGBbefore correction-(HGBchylous plasma-HGBchylous plasma ×HCTbefore correction).Results The chylemia could lead significant increase of HGB,MCH and MCHC (P<0.05).After plasma exchanging for two times,the three parameters returned to normal and the count of WBC,RBC and PLT decline slightly with no significance.There were no differences between plasma exchange method and formula calibration method.Conclusion Plasma exchange method and formula calibration method could significantly reduce the impact of chylemia on routine blood analysis,which facilitate the clinical work with correct analysis of routine blood test.
3.Establishing the combined bacterial detection procedure for the diagnosis of bacterial vaginosis
Kedi WANG ; Zhi Lü ; Jianrong SU
Chinese Journal of Laboratory Medicine 2012;(12):1167-1171
Objective To discuss the best bacterial combination for the diagnosis of Bacterial vaginosis (BV).Methods This is a retrospective study,230 BV-positive patients and 360 healthy women were enrolled based on the Amsel criteria and Nugent score.5 BV-associated bacteria,including Gardnerella vaginalis,Atopobium vaginae and Leptotrichia/Sneathia species,et al.were amplified by specific-PCR assay,the detection rate were compared between two groups.ROC curve and Kappa test were used to select the best combination.Results The detection rate of Gardnerella vaginalis,Atopobium vaginae,Leptotrichia/Sneathia species,Megasphaera species and Mobiluncus mulieris in BV group (91.3%,83.5%,39.1%,42.6% and 36.5% respectively) were markedly higher than that in healthy women (37.2%,14.4%,11.7%,8.9% and 5.6% respectively),x2 value were 168.848,275.776,60.949,92.886 and 92.68,all P < 0.05.The area under ROC curve of A.vag,G.Vag + A.vag,G.Vag + A.vag + Lepto,G.Vag + A.vag +Mega and G.Vag + A.vag + M.mul were 0.845,0.862,0.865,0.869 and 0.867,and the sensitivity and specifity were higher than 80%,the value of Kappa were larger than 0.75 (P < 0.05).Thus,these five methods were coincident.Conclusion Detection of Atopobium vaginae may be a better way for the diagnosis of BV.
4.Quantitative analysis on EB virus DNA in peripheral white blood cell,plasma and serum in patients with hematologic diseases
Kedi WANG ; Zhi LV ; Tieshan WANG ; Jianrong SU
International Journal of Laboratory Medicine 2014;(16):2161-2162
Objective To compare the load of EB virus in peripheral white blood cell,plasma and serum in the patients with he-matologic diseases,and to discuss the feasibility of detecting EB virus load by using plasma or serum instead of peripheral white blood cell.Methods Venous blood of 125 patients with hematologic diseases were collected,and peripheral white blood cell,plasma and serum were isolated.The real-time quantitative PCR was used to detect the virus load in three kinds samples with the EB virus load in peripheral blood cell as the gold standard.Results Compared to peripheral white blood,the EB virus load in plasma and ser-um showed no statistical difference(P >0.05).Conclusion Using plasma or serum instead of peripheral white blood cell for con-ducting the quantitative detection of the load of EB virus will be a reliable method.
5.Research progresses in synthetic biology of artemisinin.
Jianqiang KONG ; Wei WANG ; Kedi CHENG ; Ping ZHU
Acta Pharmaceutica Sinica 2013;48(2):193-205
Abstract: The first-line drug artemisinin is widely used against malaria. Commercially available artemisinin is extracted from plants. However, the lack of sufficient raw material, artemisinin and the cost associated with the drug's manufacture have limited the supply of ACT to most malaria sufferers in the Developing World. As such, it is important to develop a low cost, fine to environment and high-quality method to supply sufficient and reliable quantities of artemisinin in the future. The field of synthetic biology, which utilizes cell factories to manipulate microbial metabolism to enhance the production of artemisinin and its intermediates, has a particularly strong impact by providing new platforms for chemical production. After a brief introduction of the artemisinin biosynthetic pathway, the present review focuses on the introduction of artemisinin biosynthetic genes, such as the genes encoding amorpha-4, 11-diene monooxygenase, NADPH: cytochrome P450 oxidoreductase, artemisinic aldehyde delta 11(13) reductase and aldehyde dehydrogenase. The review also addresses general considerations for potential contributions of synthetic biology to artemisinin production, with an emphasis on factors influencing interest compounds production in chassis cells.
6.The advance in synthetic biology: towards a microbe-derived paclitaxel intermediates.
Wei WANG ; Yan YANG ; Xiaodong ZHENG ; Shuqiong HUANG ; Lei GUO ; Jianqiang KONG ; Kedi CHENG
Acta Pharmaceutica Sinica 2013;48(2):187-92
The synthetic biology matures to promote the heterologous biosynthesis of the well-known drug paclitaxel that is one of the most important and active chemotherapeutic agents for the first-line clinical treatment of cancer. This review focuses on the construction and regulation of the biosynthetic pathway of paclitaxel intermediates in both Escherichia coli and Saccharomyces cerevisiae. In particular, the review also features the early efforts to design and overproduce taxadiene and the bottleneck of scale fermentation for producing the intermediates.
7.Enzymatic cyclization of peptides using immobilized sortase A.
Shuxiang ZHANG ; Minzhi LIU ; Yan YANG ; Kedi CHENG ; Jianqiang KONG ; Wei WANG
Acta Pharmaceutica Sinica 2015;50(5):627-32
Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
8.Advances in functional studies of nonstructural proteins and development of antiviral agents for enterovirus 71.
Xianyun QIN ; Lin LIN ; Yan YANG ; Shuxiang ZHANG ; Jianqiang KONG ; Kedi CHENG ; Yunfeng ZHAO ; Wei WANG
Acta Pharmaceutica Sinica 2011;46(7):753-61
Human enterovirus 71 (EV71) is one of the major etiological agents for the hand, foot, and month disease (HFMD) and is causing frequent, widespread occurrence in the mainland of China. The single positive-stranded RNA genome of EV71 is translated into a single polyprotein which is autocleavaged into structural and nonstructural proteins. The functions of many nonstructural proteins characterized in the life cycle of virus are potential targets for blocking viral replication. This article reviews the studies of the structures and functions of nonstructural proteins of EV71 and the anti-enterovirus 71 drugs targeting on these nonstructural proteins.
9.Development of a yeast two-hybrid screen for selection of A/H1N1 influenza NS1 non-structural protein and human CPSF30 protein interaction inhibitors.
Jianqiang KONG ; Junhao SHEN ; Yong HUANG ; Renyu RUAN ; Bin XIANG ; Xiaodong ZHENG ; Kedi CHENG ; Wei WANG
Acta Pharmaceutica Sinica 2010;45(3):388-94
Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.
10.A cyclotide against influenza A H1N1 virus from Viola yedoensis.
Minzhi LIU ; Yan YANG ; Shuxiang ZHANG ; Liang TANG ; Huimin WANG ; Chengjuan CHEN ; Zhufang SHEN ; Kedi CHENG ; Jianqiang KONG ; Wei WANG
Acta Pharmaceutica Sinica 2014;49(6):905-12
Three cyclotides were isolated from the whole plant of Viola yedoensis in this study. The two, vary peptide E and cycloviolacin Y5, were previously reported, and a novel cycloviolacin VY1 was characterized according to the interpretation of MS/MS fragmentation of peptides which were produced from the reduced and alkylated parent peptide with the digestion of Endo Lys-C, trypsin and chymotrypsin, separately. The stability of remarkable resistance to proteolytic degradation by trypsin and chymotrypsin, and that of thermal denaturation was confirmed again. Besides, the IC50 value of cycloviolacin VYI against influenza A H1N1 virus was (2.27 +/- 0.20) microg x mL(-1). It is the first cyclotide reported with anti-influenza A H1N1 virus activity in vitro assay.