1.Application of gene chip technology to medicinal plant researches
Chinese Traditional and Herbal Drugs 1994;0(10):-
Recently,gene chip technology has become a rapidly developed biotechnology.It contains so many advantages including large-scale, high flux,and parallelism that it has been widely applied in many fields.In this paper,the updated advances on applications of gene chip technology to medicinal plant researches are discussed and the contents are ranged from isolation of differentially expressed genes,discovery of new genes,research on functional genomics,identification of Chinese materia medica,detection of genetically transformed medicinal plants,and the molecular mechanisms of medicinal plant pharmacology and their diseases as well.Some problems and prospects related to the technology are also briefly presented.
2.Sequencing of ribosomal 18S rRNA gene from Panax pseudoginseng var.notoginseng
Chinese Traditional and Herbal Drugs 1994;0(12):-
Object To identify the characteristics of 18S rRNA from the root of Panax pseudoginsengWall var. notoginseng (Burkill) Hoo et Tseng (PGSN). Methods The primers were designed according to the gene sequence of 18S rRNA from the model plant arabidopsis thaliana. 18S rRNA gene sequence of PGSN were cloned and sequenced and compared with that of the model plant A. thaliana and P. pseudoginseng subsp.Wall himalacus var. angustifolius. Results Part of the characteristics of ribosomal 18S rRNA gene sequence of PGSN from Jingxi, Guangxi Province were identified, which revealed that the 18S rRNA gene sequence of PGSN was 98% similar to that of P. pseudoginseng subsp. himalalaicus var . angustifolius and 96% similar to that of the model plant A. thaliana. Conclusion The use of informations obtained from the model plant, A. thaliana may promote the research progress of molecular biology of TCM drugs.
3.The Regulative Role of ERK5/Bim Pathway in Hypothermal Stimulation Induced Neonatal Rat’s Cardiomyocytes Injury and Apoptosis
Yaosheng WANG ; Xiaoshu CHENG ; Kui HONG ; Zonggui WU ; Yigang LI
Chinese Circulation Journal 2014;(7):540-544
Objective: To explore the regulative role of extracellular regulated protein kinase-5 (ERK5)/Bcl-2 interacting mediator of cell death (Bim) pathway in hypothermal stimulation induced neonatal rat’s cardiomyocytes (CMs) damage and apoptosis.
Methods: CMs were cultured for hypothermal stimulation and the speciifc siRNA was used to down-regulate the ERK5 or Bim in CMs. The cell apoptosis was detected by lfow cytometry, protein expression was examined by Western blot analysis, the intracellular Ca2+, reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were evaluated by lfuorescent labeling and lfow cytometry.
Results: In hypothermal stimulated CMs, ERK5 siRNA could promote Bim protein expression, but Bim siRNA could not inlfuence ERK5, while attenuated p-ERK5 expression. ERK5 siRNA induced higher apoptosis rate, while Bim siRNA could decrease such effect. ERK5 siRNA increased the intracellular Ca2+overloading, ROS activation andΔΨm damage, while Bim siRNA played the role to against those effects in hypothermal stimulated CMs.
Conclusion: Our study revealed that ERK5/Bim pathway played the important regulative roll in hypothermal stimulation induced neonatal rat’s CMs damage and apoptosis.
4.Genetic cloning and sequence analysis of farnesyl pyrophosphate synthase in Panax notoginseng
Li CHEN ; Xiuwan LAN ; Shen LI ; Hua ZHU ; Yaosheng WU
Chinese Traditional and Herbal Drugs 1994;0(07):-
Objective To clone and sequence the cDNA encoding farnesyl pyrophosphate synthase(FPS) from Panax notoginseng.Methods The cDNA,encoding FPS in P.notoginseng,was amplified by RACE strategy with the total RNA of root as the template.The fragment of FPS was cloned and sequenced.Results The analysis results revealed that the full-length cDNA had(1 409) bp with an open reading frame encoding 343 amino acids of protein.The FPS sequence had 95%,87%,and 86% amino acid sequence homology to the FPS sequence of Centella asiatica,Parthenium argentatum,and Artemisia annua,respectively. Conclusion The cDNA encoding FPS from P. notoginseng is cloned and reported.This works provide a foundation for exploring the mechanism of saponins biosynthesis and application to the other medical plants.
5.Multiband mucosectomy in treatment of early cancer and precancerous lesions of esophagogastric junction
Bin DENG ; Dacheng WU ; Mei WANG ; Yaosheng CHEN ; Yuanzhi WANG ; Jian WU ; Yanbing DING
Chinese Journal of General Practitioners 2014;13(12):1014-1016
Eighteen patients with early cancer and precancerous lesions of esophagogastric junction underwent multiband mucosectomy (MBM) in Yangzhou First People's Hospital from January 2013 to February 2014.The clinicopathological data of patients were analyzed retrospectively and the short-term efficacy and safety of MBM were evaluated.Operations were successful in all 18 cases.The mean operative time was 35.8 min.Intraoperative bleeding occurred in 3 cases and successfully managed by endoscopic hemostasis.Small perforation occurred in 1 patient and was closed by metal clips.Pathological examination showed mucosal cancer in 5 cases,high-grade intraepithelial neoplasia in 8 cases and low-grade intraepithelial neoplasia in 5 cases.No relapse of cancer was found during follow-up.Results indicate that MBM is an effective and safe method in treatment of early cancer and precancerous lesions in the esophagogastric junction.
6.Expression, purification and function of hVEGF_(121)/EGFP fusion protein in bone marrow-derived mesenchymal stem cells
Lipeng HE ; Yaosheng WANG ; Yihua ZHOU ; Yu JIANG ; Xiaoling WU ; Xiaoshu CHENG
Chinese Journal of Tissue Engineering Research 2009;13(49):9731-9734
OBJECTIVE: To purify hVEGF_(121)/EGFP fusion protein using transfected BMSCs as culture media, in addition, to detect the function of hVEGF_(121)/EGFP fusion protein in vitro.METHODS: The pEGFP-N_2-hVEGF_(121) recombinant plasmid, which was constructed in the preliminary work of our study group,was used to extract the plasmid DNA. BMSCs were transfected with pEGFP-N2-hVEGF_(121) by positive ionic liposome transfection method. Under a fluorescent microscopy, the expression of hVEGF_(121)/EGFP fusion protein was detected. The hVEGF_(121)/EGFP fusion protein was purified with Am icon ultrafiltration centrifuge tube and the expression of fusion protein was detected by Western-Blotting method.RESULTS: The BMSCs, which transfected with pEGFP-N2-hVEGF_(121), was observed under the fluorescent microscope. Western blotting confirmed that pEGFP-N_2-hVEGF_(121) fusion protein expressed in the culture media of transfected BMCS. MTT results showed the number of human umbilical vein endothelial cells in the fusion protein team was significantly greater than that in the control group (P < 0.05), and Miles test confirmed that pEGFP-N_2-hVEGF_(121) fusion protein increased the permeability of the blood vessel wall.CONCLUSION: ①This study successfully confirmed the pEGFP-N_2-hVEGF_(121) recombinant plasmid, which carrying VEGF_(121)/EGFP fusion protein, can be expressed in BMSCs.②The VEGF_(121)/EGFP fusion protein have the function of wild-type VEGF in vitro.
7.The variety of the neutrophil volume and cytoplasm-nucleus complex and their significance in acute bacterial infection and stringency diseases
Yanhui JIN ; Mingshan WANG ; Lihong YANG ; Yaosheng XIE ; Zhuo ZHANG ; Xiuji WU
Chinese Journal of Laboratory Medicine 2009;32(3):326-330
Objective To investigate the variety and clinical value of the neutrophil volume and cytoplasm-nucleus complex in patients with bacterial infection, cardiovascular or cerebrovascular accident and major surgery operation. Methods 125 patients with bacterial infection, 64 patients with acute cardiovascular or cerebrovascular accident, 66 patients after major surgery operation and 69 normal subjects were selected in the study. Total WBC counts (WBC), percentage of neutrophils (NE), and the VCS parameters of neutrephils including the mean channels of cell volume ( NEV), conductivity ( NEC), light scatter(NES) and the SD of these parameters( NEVSD, NECSD, NESSD)were measured by automatic blood cell analyzing instrument. The sensitivity and specificity of the WBC, NE and the VCS parameters of neutrophils were analyzed with receive operating characteristic (ROC) curve. Results The levels of NEV, NES and NEVSD in acute bacterial infection group were 154.3 ± 15.2, 135.7 ± 9.9, 26.8 ± 4.2 respectively. The levels of NEV, NES and NEVSD in post major surgery operation group were 147.2±8.9, 141.5 ± 7.7, 23.0 ± 2. 8 respectively. The levels of NEV, NES and NEVSD in acute cardiovascular or cerebruvascular accident group were 144.9 ± 5. 2, 146.0 ±5.0, 19. 6±1.6 respectively. The levels of NEV, NES and NEVSD in healthy control group were 139.7±4.6, 145.0±3.8, 18.2±1.3 respectively. The differences of these parameters among these groups had statistical significance ( F = 17. 650, 38. 122, 54. 604,P<0. 05). And the changes of NEV, NES and NEVSD in bacterial infection group were most obvious among those three groups. The levels of NEV, NES and NEVSD were 146.5±9.5, 144.3 ± 9.4, 21.3 ± 3.3 respectively in stress diseases groups which included acute cardiovascular or cerebrovascular accident group and post major surgery operation group. The differences of these parameters between stress diseases group and acute bacterial infection group had statistical significance ( t = - 2.840, 7.533, - 8.999,P<0.01). The areas under the ROC curve of NEVSD, NEC, NES and NECSD were 0.893, 0. 845, 0. 833 and 0. 849 respectively. The sensitivity of 83. 3% and specificity of 82. 0% could be achieved by selecting the cut-off equal to or greater than 24. 0. Conclusions The variety of neutrophil volume, nuclear size and cytoplasmic granularity changed obviously in patients with acute bacterial infection and common stress diseases, and the variety in acute bacterial infection is more obvious than that in common stress diseases. The sensitivity and specificity of VCS parameters of neutrophils are higher than those of WBC or NE for predicting infection, and the NEVSD is the most predictable indicator of acute bacterial infection.
8.Blade differentiation and plantlet regeneration of Gynostemma pentaphyllum
Ruiqiang ZHAO ; Zhengzhu LING ; Yu LUO ; Xiongying LI ; Junfu JIANG ; Yaosheng WU
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective To construct a stably-performing and high-efficiency regeneration system of Gynostemma pentaphyllum and lay a foundation for the gene transformation of it.Methods The blades of five-leaf G.pentaphyllum were used as the explants and cultured in MS media with different portions of hormone to induce fascicled-bud,root and plantlet regeneration.Results The medium suitable for inducing the blade differentiation of G.pentaphyllum was MS+6-BA 1.0 mg/L and for the frequency of blade differentiation could reach 40%.The cultural medium MS+6-BA 1.0 mg/L was suitable for the sub-multiplication of fascicled-bud and the medium 1/2 MS for root inducement and the plantlet regeneration.Conclusion A stably-performing acceptor system of the direct differentiation for Agrobacterium tumefaciens mediated gene transformation of G.pentaphyllum blades is constructed.
9.Survey of clinicians' needs of support for conducting clinical research
Guangyu CHEN ; Xuehai LI ; YaoSheng WANG ; Xi ZHANG ; Weijie WU ; Rong LUO ; Wei ZHANG ; Zhongmin ZHENG ; Kun SUN ; Xin ZHOU
Chinese Journal of Medical Science Research Management 2020;33(1):60-63
Objective To explore and understand limitations and needs of support for conducting clinical research among clinicians in China.Methods The in-service clinician in our hospital were enrolled in as survey participants,survey questionnaires were designed and filled up by target audience in the questionnaire star platform.Results Main problems identified as restrictions for conducting clinical studies including lack of data platform,have ideas but not protocols,lack of research funding and project management.On the other hand,identified needs for support including research-related technical support,project management facilitation,training,external communication and exchange,as well as quality risk management.Conclusions Hospital should establish Clinical Research Unit to help clinicians to improve research quality by training,providing technical support and administrative service of clinical research.
10.Practical exploration of clinical research unit platform construction and management mode based on research-oriented hospitals:The case study of a tertiary hospital in Shanghai
Rong LUO ; Xi ZHANG ; Yaosheng WANG ; Guangyu CHEN ; Weijie WU ; Xuehai LI ; Xin ZHOU
Chinese Journal of Medical Science Research Management 2019;32(2):81-85
Objective To summarize the practical experiences of Clinical Research Unit (CRU) platform established by Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine,provide reference for domestic institutions that have prepared or to prepare to establish CRU platform.Methods Oriented by demand,this platform has set up six modules including project management module,methodology support module,data management module,quality risk management module,project transformation module and training and international exchange module.Besides,it also establishes and improves the corresponding module system,quality assurance,technical support and information data platform system.Results With this platform,the enthusiasm of departments to carry out clinical research has been continuously enhanced,the talent pool of clinical research has been expanded;the awareness of clinical research cooperation has been improved and the international cooperation fields have been further extended;the number of clinical research projects and funding have increased rapidly.Conclusions CRU Platform in Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine provides all-dimensional technical support and whole-process informatization management to improve clinical research quality and efficiency.While learning from the advanced international concepts,we still need to explore and improve the platform construction and management mode in line with general research-oriented hospitals in China,further to enhance the academic research internationally.