2.The difference of T lymphocyte subsets between mild type and severe type in patients infected with the 2009 pandemic influenza A (H1N1) virus and its clinical significance
Jun WANG ; Jun JIN ; Fang HUANG ; Jianan HUANG ; Cheng JI ; Yueping SHEN ; Hua XU
Chinese Journal of Emergency Medicine 2011;20(2):193-197
Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P<0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P < 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P < 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.
3.The culture and differentiation of adult bone marrow-derived pluripotential mesenchymal stem cells into insulin-producing cells
Lihong CHEN ; Hua CHENG ; Jun MIN ; Muchao WU ; Li YAN ; Zuzhi FU
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To study the culture and characteristics of mouse adult bone marrow-derived pluripotential mesenchymal stem cells and its potential to differentiate into insulin secretion cells. METHODS: Cells were plated on 60% DMEM-LG and 40% MCDB-201 medium supplemented with 2% fetal calf serum and 10 ?g/L PDGF-BB, 10 ?g/L EGF and 1?10~6 U/L LIF. The proliferation rate, phenotype and oct-4 mRNA were tested. After it was plated on serum-free medium DMEM/F12 with GLP-1 and nicotinamide, the nkx2.2 ngn3, pdx-1 and insulin 2 mRNA were tested. RESULTS: The cells were round with large nucleus and scant cytoplasma. They were CD13~+, CD44~-, CD45~- and MHCⅡ~-. Oct-4 mRNA were present. The nkx2.2 pdx-1 and insulin 2 mRNA were presented in cells plated on the inducing medium at 14 days. CONCLUSION: The adult bone marrow-derived pluripotent stem cells were cultured and they has the possibilities to be induced into insulin-secreting cells.
4.Statin in the treatment of ALI/ARDS: a systematic review and Meta-analysis basedon international databases
Mingqi CHEN ; Jun LU ; Qiuhua CHEN ; Lu CHENG ; Yanxia GENG ; Hua JIANG ; Xing WANG
Chinese Critical Care Medicine 2017;29(1):51-56
Objective To confirm the effects of statin therapy on mortality of patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS). Methods PubMed/Medline, Embase, Web of Science andCochrane Central Register of Controlled Trials were searched for articles using the terms acute lung injury, ALI,acute respiratory distress syndrome, ARDS, statin, simvastatin and rosuvastatin updated to November 17,2015. Randomized controlled trial (RCT) or observational cohort studies investigating the effects of statin therapy onmortality in patients with ALI or ARDS were all identified, without date or language restriction. The control group wasgiven conventional treatment, while the experimental group was treated with statins additionally. The primary outcomewas in-hospital mortality. Meanwhile, ventilator-free day, intensive care unit (ICU)-free day, ICU length of stay (LOS)and ICU mortality were also analyzed. RevMan 5.2 and STATA 13 software were used for systematic review and Metaanalysis, and funnel plot was used to analyze the publication bias. Results A total of five trials including threerandomized controlled trials and two observational studies were included. Among 1636 patients enrolled in the study,there were 739 patients in experimental group, and 897 in control group. It was shown by Meta analysis that there was nosignificant difference in in-hospital mortality between experimental group and control group [relative risk (RR) = 0.96,95% confidence interval (95%CI) = 0.79-1.15, P = 0.63]. The subgroup analysis based on RCT and cohort study, or thesubgroup analysis of different statins showed that there was no significant difference in in-hospital mortality betweenthe experimental group and the control group (both P > 0.05). There were no significant differences in ventilator-freedays [mean difference (MD) = 1.41, 95%CI = -0.32-3.13, P = 0.11], ICU-free days (MD = -0.23, 95%CI = -1.61-1.15,P = 0.75), ICU length of stay (MD = -1.03, 95%CI = -6.55-4.50, P = 0.72), or ICU mortality (RR = 0.88, 95%CI =0.68-1.14, P = 0.33) between the experimental group and the control group. It was shown by funnel plot that there was nopublication bias in in-hospital mortality. Conclusion The systematic review and meta-analysis suggests that statin may not be associated with a significant reduction in mortality, ventilator-free day, ICU-free day and ICU length of stayin patients with ALI/ARDS.
6.Study on in vitro metabolic rate and metabolites or 9-dehydro-17-dehydro-andrographolide.
Jun SHAO ; Wei-kang CHEN ; Dong-kun ZHENG ; Shuang-cheng MA ; Yue-hua LUO
China Journal of Chinese Materia Medica 2015;40(5):971-977
To investigate the metabolic rate and metabolites of 9-dehydro-17-dehydro-andrographolide, which is the main active ingredient in Xiyanping injection, by using the in vitro rat liver microsome incubation system. 9-dehydro-17-dehydro-andrographolide was incubated together with liver microsome mixed with NADPH. Its metabolic rate was studied by determining its residual concentrations with the UHPLC-MS/MS method; Its metabolites were identified by the UPLC-TOF-MS(E) method. The results showed that 9-dehydro-17-dehydro-andrographolide was metabolized faster than rat liver microsomes mixed with coenzymes, with t½ and CL of (19.7 ± 0.5) min and (35.1 ± 0.8) mL x min(-1) x g(-1) (protein), respectively. Based on the high resolution mass spectrum data and information from literatures, altogether nine metabolites of 9-dehydro-17-dehydro-andrographolide were identified in the incubation system, particularly hydroxylated and dehydrogenized products. The results of identification would provide a basis for screening out more active andrographolide derivatives.
Animals
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Chromatography, High Pressure Liquid
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Diterpenes
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chemistry
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metabolism
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Drugs, Chinese Herbal
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chemistry
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metabolism
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Microsomes, Liver
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chemistry
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metabolism
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Molecular Structure
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Rats
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Tandem Mass Spectrometry
8.Establishment of prescription research technology system in Chinese medicine secondary exploitation based on "component structure" theory.
Xu-Dong CHENG ; Liang FENG ; Jun-Fei GU ; Ming-Hua ZHANG ; Xiao-Bin JIA
China Journal of Chinese Materia Medica 2014;39(21):4272-4276
Chinese medicine prescriptions are the wisdom outcomes of traditional Chinese medicine (TCM) clinical treatment determinations which based on differentiation of symptoms and signs. Chinese medicine prescriptions are also the basis of secondary exploitation of TCM. The study on prescription helps to understand the material basis of its efficacy, pharmacological mechanism, which is an important guarantee for the modernization of traditional Chinese medicine. Currently, there is not yet dissertation n the method and technology system of basic research on the prescription of Chinese medicine. This paper focuses on how to build an effective system of prescription research technology. Based on "component structure" theory, a technology system contained four-step method that "prescription analysis, the material basis screening, the material basis of analysis and optimization and verify" was proposed. The technology system analyzes the material basis of the three levels such as Chinese medicine pieces, constituents and the compounds which could respect the overall efficacy of Chinese medicine. Ideas of prescription optimization, remodeling are introduced into the system. The technology system is the combination of the existing research and associates with new techniques and methods, which used for explore the research thought suitable for material basis research and prescription remodeling. The system provides a reference for the secondary development of traditional Chinese medicine, and industrial upgrading.
Drug Prescriptions
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Medicine, Chinese Traditional
9.Establishment of industry promotion technology system in Chinese medicine secondary exploitation based on "component structure theory".
Xu-Dong CHENG ; Liang FENG ; Ming-Hua ZHANG ; Jun-Fei GU ; Xiao-Bin JIA
China Journal of Chinese Materia Medica 2014;39(20):4065-4069
The purpose of the secondary exploitation of Chinese medicine is to improve the quality of Chinese medicine products, enhance core competitiveness, for better use in clinical practice, and more effectively solve the patient suffering. Herbs, extraction, separation, refreshing, preparation and quality control are all involved in the industry promotion of Chinese medicine secondary exploitation of industrial production. The Chinese medicine quality improvement and industry promotion could be realized with the whole process of process optimization, quality control, overall processes improvement. Based on the "component structure theory", "multi-dimensional structure & process dynamic quality control system" and systematic and holistic character of Chinese medicine, impacts of whole process were discussed. Technology systems of Chinese medicine industry promotion was built to provide theoretical basis for improving the quality and efficacy of the secondary development of traditional Chinese medicine products.
Chemistry, Pharmaceutical
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economics
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standards
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China
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Drug and Narcotic Control
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economics
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Drugs, Chinese Herbal
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chemistry
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economics
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standards
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Humans
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Medicine, Chinese Traditional
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economics
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standards
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Quality Control
10.Establishment and optimization of real-time PCR with SYBR Green Ⅰ for quantification of survivin
Li-Hua HU ; Zheng-Jiang CHENG ; Yi-Rong LI ; Shao-Jun HUANG ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.