2.Detection and purification of cultured rat retinal Müller cell by flow cytometer
Min, GONG ; Bo-lin, XIE ; Shu-qing, LI ; Fan, LI
Chinese Journal of Experimental Ophthalmology 2011;29(2):121-124
Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.
4.Effects of inhibition of Cripto gene siRNA on vascular endothelial growth factor of colon cancer cell line LS-174T
Yu FAN ; Youli ZHANG ; Hua LI ; Zefeng XU ; Shu ZHENG
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To study the effects of Cripto gene on vascular endothelial growth factor(VEGF) of colon carcinoma cells.METHODS: Cripto siRNA was designed and constructed.Colon cancer LS-174T cells were divided into 4 groups: control group and different dose (3.125,6.25 and 12.5 nmol/L) of siRNA groups.After transfected for 24,48 and 72 h,colon cancer cells were harvested to carry on the next tests.Expression of Cripto mRNA was determined with real-time PCR,and immunofluorescence isothiocyanate(FITC) labeling assay and Northern blotting were performed to examine the expression of protein and mRNA of VEGF,respectively.The cells in control group and cells transfected with 12.5 nmol/L siRNA were inoculated into nude mice respectively.30 days after inoculated,the mice of two groups were executed,and immunohistochemical(ICH) assay was used to evaluate the VEGF protein of mice tumor.RESULTS: siRNA down-regulated the Cripto mRNA in a dose and time dependent manner.Protein and mRNA of VEGF in transfected cells reduced in a dose and time dependent manner.Compared to control,the expression of VEGF protein from ICH assay was lowered significantly(P
7.Regularity of drugs compatibility of anti-hepatoma traditional Chinese medicine ancient prescriptions and risk evaluation of anti-hepatoma new drug research and development.
Jing ZHANG ; Hong-Fa LI ; Wei FAN ; Zhen LIU ; Shu-Li MAN ; Shu-Yong SI ; Wen-Yuan GAO
China Journal of Chinese Materia Medica 2014;39(19):3870-3875
Traditional Chinese ancient prescriptions have been used for treatment of liver cancer for a long history and the scientific and rational compatibility is a great wealth for modern research and development (R&D) of new drugs. The research and development of new drugs are often accompanied with a large investment, a long cycle and a high risk, especially for the anti-tumor drugs R&D which are facing more risks and lower successful rate. In this research, the regularity of compatibility of drugs was analyzed from 124 anti-hepatoma ancient prescriptions by computer program. The results can offer help to the R&D of anti-hepatoma new drugs and reduce the risk of drug screening. In addition, we surveyed 22 companies in this field from six provinces such as Beijing, Shanghai, Tianjin and so on and obtained 240 risk assessment questionaires. Then we used qualitative analysis method to interpret the greatest impacts for the risks in the process of R&D, production and sales of anti-hepatoma new drugs. The study provides a basis for anti-liver cancer drugs R&D researchers, who can take effective measures to reduce the R&D risks and improve successful rate.
Carcinoma, Hepatocellular
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drug therapy
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history
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China
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Drug Discovery
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history
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Drug Incompatibility
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Drug Prescriptions
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history
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Drugs, Chinese Herbal
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history
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therapeutic use
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History, Ancient
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Humans
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Liver Neoplasms
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drug therapy
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history
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Research
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history
8.Primary culture and purification of cerebral astrocyte of tree shrew.
Min GONG ; Shu-qing LI ; Fan LI
Acta Physiologica Sinica 2011;63(1):89-92
The aim of this study is to establish a method to culture and purify cerebral astrocyte of tree shrew (Tupaia belangeri), a kind of new laboratorial animal which is a relative of primates. Newborn tree shrews were used in this experiment. The cortex of cerebrum was isolated and placed in 4°C for 20 min to injure neurons. The cortical tissue was disaggregated by trypsin digestion. Differential attachment method was used to remove fibroblasts. The mixed culture was rinsed by trypsin (0.005%) solution to remove neurons. Upon reaching 70% confluence, the culture was subjected to static trypsin digestion until a white slice film exfoliated from the bottom of culture bottle. This film, i.e. astrocyte layer, was taken out and cultured, and the third passage was identified by immunocytochemical staining and immunofluorescence with anti-glial fibrillary acidic protein (GFAP) antibody. The result showed the purity of tree shrew astrocytes was more than 98%. Thus the method to culture highly purified astrocyte of tree shrew was successfully established, which would contribute to further study in central nervous system physiology and diseases in this new laboratorial animal.
Animals
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Astrocytes
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cytology
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Brain
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cytology
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Cell Separation
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methods
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Primary Cell Culture
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methods
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Tupaiidae
9.Dynamic monitoring risk of anti-hepatoma new drug development.
Jing ZHANG ; Wei FAN ; Hong-Fa LI ; Shu-Li MAN ; Zhen LIU ; Wen-Yuan GAO
China Journal of Chinese Materia Medica 2014;39(20):4050-4053
Risk monitoring of new Chinese patent anti-hepatoma drugs is tracking recognized risks and residual risks, identifying emerging risk and ensure the implementation of the plan, estimating the process of reducing effectiveness. The paper is mainly through understanding the status of Chinese patent anti-hepatoma drugs, the content, characteristic and analysis method of dynamic risk monitoring, and then select the risk control indicators, collect risk information. Finally, puts forward the thought of anti-hepatoma drugs listed evaluation in our country, and try to establish the model of dynamic risk management of anti-hepatoma drugs.
Antineoplastic Agents, Phytogenic
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adverse effects
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economics
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therapeutic use
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Carcinoma, Hepatocellular
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drug therapy
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Drug Discovery
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economics
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legislation & jurisprudence
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organization & administration
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Drug and Narcotic Control
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economics
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legislation & jurisprudence
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organization & administration
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Drugs, Chinese Herbal
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adverse effects
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economics
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therapeutic use
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Humans
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Liver Neoplasms
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drug therapy
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Product Surveillance, Postmarketing
10.Risk Factors of Central Venous Catheter Related Sepsis in Total Parenteral Nutrition: Analysis of Logistic Regression
Shu-Shan FAN ; Zhao-Ju LV ; Shou-Guo ZHAO ; Jian-Zhong CHEN ; Xia LI ; Jin ZHANG ;
Chinese Journal of Nosocomiology 1994;0(01):-
OBJECTIVE To analyze risk factors in order to provide scientific gist in diagnosis and treatment of central venous catheter related sepsis(CRS) in patients with total parenteral nutrition(TPN). METHODS To make comparison of the 57 cases of CRS with 423 cases of non-CRS during 1998-2002.First,monovariable chi-square test and then non-condition Logistic regression analysis of the markedly different factors in SPSS10.0 were conducted. RESULTS The major risk factors might be infectious disease,duration of central venous catheter in,location of catheter, type of catheter and material of catheter,and serum protein