1.Effect of Wenyang Huoxue Lishul Recipe Containing Serum on Expression of Cathepsin L in Puromycin Aminonucleoside-induced Injury of Mouse Glomerular Podocytes.
Wen-wen QIU ; Jun YUAN ; Liu YANG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(5):602-607
OBJECTIVETo observe the effect of Wenyang Huoxue Lishui Recipe (WHLR) containing serum on the expression of cathepsin L (CatL) in puromycin aminonucleoside-induced injury of mouse glomerular podocytes.
METHODSMouse podocyte cells (MPCs) in vitro cultured were divided into the normal control group, the model group, the dexamethasone (DEX) group, 10% WHLR containing serum group, 20% WHLR containing serum group, the vehicle serum control group. MPCs in the normal control group were cultured at 37 degrees C culture solution for 24 h. 45 mg/L puromycin was acted on MPCs in the model group for 24 h. On the basis of puromycin intervention, 1 limol/L DEX was co-incubated in MPCs of the DEX group for 24 h; 10% or 20% WHLR containing serum was co-incubated in MPCs of the 10% WHLR containing serum group and 20% WHLR containing serum group for 24 h. The vehicle serum control group was also set up by incubating with WHLR containing serum alone for 24 h. The expression of CatL and its substrate Synaptopodin in podocytes were detected by cell immunofluorescence staining. FITC-conjugated phalloidin was used to stain F-actin. A cortical F-actin score index (CFS index) was designed to quantify the degree of cytoskeletal reorganization in cultured podocytes.
RESULTSCompared with the normal control group, the expression of synaptopodin significantly decreased and the expression of CatL significantly-increased in the model group. F-actin arranged in disorder, gradually forming pericellular F-actin ring. CFS index was obviously elevated (P < 0.01). Compared with the model group, the epression of synaptopodin increased, the expression of CatL decreased, and CFS index also decreased in the DEX group, 10% WHLR containing serum group, and 20% WHLR containing serum group (P < 0.05, P < 0.01). Compared with the DEX group, the expression of synaptopodin decreased in 10% WHLR containing serum group, CFS index also decreased in 20% WHLR containing serum group (P < 0.05).
CONCLUSIONSWHLR could up-regulate the expression of synaptopodin, down-regulate the expression of CatL, and alleviate cytoskeletal reorganization of F-actin. It was helpful to stabilize the cytoskeleton of F-actin and improve the merging of podocytes.
Actins ; metabolism ; Animals ; Cathepsin L ; metabolism ; Cells, Cultured ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Kidney Glomerulus ; cytology ; Mice ; Microfilament Proteins ; metabolism ; Podocytes ; drug effects ; pathology ; Puromycin Aminonucleoside ; adverse effects ; Up-Regulation
2.Surgical Treatment of Advanced Rectal Carcinoma in Aged People
Shaohua YANG ; Shaoyi CHEN ; Jun WEN
Journal of Chinese Physician 2001;0(04):-
Objective To explore the surgical treatment of advanced rectal carcinoma in aged people. Methods The clinical data of 65 aged patients with advanced rectal carcinoma from 2000 to 2004 were analyzed retrospectively. Results All cases were Dukes D stage or accompanied with hepatic metastasis. Hartmann's operation was performed in 26 cases, single lumen sigmoidostomy in 11 cases, double lumen sigmoidostomy in 11 cases and resection of hepatic metastatic cancer in 12 cases. Rectal upper arterial catheter chemotherapy was performed in 19 cases and hepatic arterial catheter chemotherapy in 15 cases. The 3-year survival rate of Hartmann's operation was 34%(9/26), and the average survival time was 20?3.17 months in the patients underwent rectal upper and hepatic arterial catheter chemotherapy, and 10?1.77 months in the patients who could not be treated surgically(P
3.Determination of Ginsenoside Rb_1,Ginsenoside Re and Ginsenoside Rg_1 in Yixin Fumai Granula by HPLC
Jun YANG ; Shihui WU ; Xiao WEN
China Pharmacy 2007;0(30):-
OBJECTIVE: To determine Ginsenoside Rg1,Ginsenoside Re and Ginsenoside Rb1 in Yixin fumai granula by HPLC.METHODS: Samples were determined on a Diamond C18(150 mm?4.6 mm,5 ?m).The mobile phase consisted of accetonitrile-water by gradient elution with flow rate at 1 mL?min-1,UV detection wavelength at 203 nm,column temperature at 25 ℃ and sample size at 10 ?L.RESULTS: The linear ranges of Ginsenoside Rg1,Ginsenoside Re and Ginsenoside Rb1 were 0.645~6.450 ?g(r=0.999 8),0.54~5.40 ?g(r=0.999 7) and 0.605~6.050 ?g(r=0.999 9),respectively;the average recoveries were 100.59%(RSD=2.03%),98.70%(RSD=1.46%)and 98.99%(RSD=1.19%),respectively.CONCLUSION: The method is sensitive,simple and accurate,and it can be used for the quality control of Yixin fumai granula.
4.Surface labeling of bone marrow mesenchymal stem cells by biotin-streptavidin
Lin YANG ; Fuli LUO ; Yun LI ; Jun WEN ; Yang XU
Chinese Journal of Tissue Engineering Research 2016;20(10):1382-1388
BACKGROUND:Currently, there is a lack of efficient, non-invasive way to transplant stem cels to the target organ or tissue. Exploring a way to guide targeting transplantation of stem cels and to improve the efficiency of stem cel homing is now one of focuses in the field of stem cels research.
OBJECTIVE: To establish a simple and feasible method to chemicaly modify the cel surface using biotin-streptavidin reaction system, and to evaluate the efficiency of this method to label bone marrow mesenchymal stem cels (BMSCs) and its effects on cel biological functions.
METHODS: Passage 3 BMSCs were obtained by whole bone marrow culture method and verified by flow cytometry. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated LewisX (SLeX), to the BMSCs surface. The labeling rate of BMSCs was assessed using fluorescence microscope, the vitality of BMSCs was evaluated by trypan blue staining, and the proliferation of BMSCs was evaluated by cel counting kit-8 assay. Adipogenic and osteogenic inductions were used to evaluate the effect of the method on the multi-differentiation function of BMSCs.
RESULTS AND CONCLUSION: After culture for 2 weeks, passage 3 BMSCs were obtained and confirmed by expressing CD90, CD29 and lack of CD34, CD45. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were successfuly used to equip sialyated LewisX (SLeX) to the BMSCs surface and had minor effects on the vitality, proliferation, and differentiation of BMSCs. This method was simple for surface modification and had a high modification rate of 88%. The homing of BMSCs modified by this method to the target organ or tissue could be greatly enhanced. Therefore, this method potentialy could have extensive and important applications.
5.Application of RNA Secondary Structure in Phylogenetic Analysis of Microbiology
Yang LIU ; Xiao-Long CUI ; Wen-Jun LI ; Qian PENG ;
Microbiology 1992;0(02):-
Attention was gradually paid by biologists to the using of RNA secondary structure in the classification of microbiology and phylogenetic relationship analysis in recent years. The development around the research was summarized here briefly. And more emphasis was given to the part introducing the application of RNA secondary structure to the analysis of phylogenetic relationship.
6.Experimental study of the protective effect of endotoxin pretreatment on hippocampal neurons in rat forebrain following ischemia-reperfusion
Qiang XIANG ; Jun YANG ; Liang WEN ; Minghua LIU ;
Journal of Third Military Medical University 2003;0(19):-
Objective To investigate the protective effect of endotoxin pretreatment on hippocampal neurons in rat forebrain following ischemia reperfusion and its possible mechanism. Methods Rat forebrain ischemia reperfusion model was used. The effects of endotoxin pretreatment on the changes of superoxide dismutase (SOD), glutathione peroxidase (GSH PX) and malondialdehyde (MDA) activities and the neuron count in CA1 region were observed. Results Pretreament with endotoxin before cerebral ischemia enhanced the activities of SOD and GSH PX but decreased MDA level and the number of ischemic neurons in CA1 region. Conclusion Endotoxin pretreatment can protect the neurons in rat forebrain against ischemia reperfusion injury. The mechanism may be associated with the enhancement of endogenous antioxidant activity in central nervous system.
7.Peripheral blood stem cell transplantation in the treatment of dilated cardiomyopathy: A 3-month follow-up in 42 cases
Tong ZHANG ; Chao LIU ; Jun YANG ; Xiang ZHANG ; Wen LI
Chinese Journal of Tissue Engineering Research 2007;0(16):-
AIM: Recent studies showed that stem cells could replace injured cardiomyocyte and increase the number of functional cardiomyocytes. Researching the pertinent literature in China Journal Full-text Database (CJFD) published between 2005 and 2008 indicated that the researches on stem cell transplantation in the treatment of primary dilated cardiomyopathy were few. This study investigated the therapeutic efficacy and security of peripheral blood stem cell transplantation in coronary artery in treatment of dilated cardiomyopathy and its effects on left ventricular function. METHODS: Forty-two patients with dilated cardiomyopathy between December 2006 and September 2007 were enrolled at the Department of Cardiology of First People's Hospital of Yunnan, including twenty-eight males and fourteen females, averagely aged (56?3) years. Inclusive criteria: patients with less than 65 years, left ventricular enlargement, and left ventricular ejection fraction (LVEF) ≤ 45%, and without coronary artery disease after coronary arteriongraphy. Informed consents were obtained from patients. Patients were divided into stem cell transplantation group (n=15) and control group (n=27) on the basis of whether being treated by stem cell transplantation. Patients in the stem cell transplantation group were consecutively administered granulocyte colony-stimulating factor (G-CSF) by hypodermic injection to stimulate bone marrow stem cells themselves based on conventional treatment for five days. Peripheral blood stem cell suspension was disassociated on the 6th day, and the collected suspension was injected into left anterior descending branch over the wire saccule tube for autologous peripheral blood stem cell transplantation. Patients in the control group were administered by conventional treatment of dilated cardiomyopathy. Security and adverse reaction were observed during the mobilization, collection and returning injection of peripheral blood stem cell by coronary artery. Morphous, cardiac function and motion index of left ventricle wall were evaluated using ultrasoundcardiogram before and 3 months after transplantation. Survival rate and incidence rate of heart incidents were compared. RESULTS: Three months after stem cell transplantation in coronary artery, there were a significant decrease in cardiac end-systolic volume (ESV), cardiac end-diastolic volume (EDV) and motion index of left ventricle wall, but a significant increase in LVEF(P
8.CYP2D6 genotypes and phenotypes in Chinese Han, Uygur and Kazakh populations.
Guochang LI ; Jun YANG ; Ting ZHOU ; Zhigang CHEN ; Wen CHEN
Acta Pharmaceutica Sinica 2011;46(2):238-42
This study is to compare the influence of CYP2D6 *3 and *4 genotypes and phenotypes on the metabolic activity of CYP2D6 in Chinese Han, Uygur and Kazakh ethnic groups. Allele specific amplification (ASA) was used to determine the CYP2D6*3 and CYP2D6*4 genotypes. Phenotypes of CYP2D6 in all subjects were determined using dextromethorphan as probe drug by HPLC methods. Among the 132 Han subjects, one subject (0.76%) exhibited the *1/*3 combination, and one (0.76%) exhibited the *1/*4. Among the 136 Uygur subjects, 4 subjects (2.94%) showed the *1/*3 combination, 12 (8.82%) showed *1/*4, 4 (2.94%) showed *4/*4, and one (0.74%) showed *3/*4. Among the 116 Kazakh subjects, 2 (1.72%) exhibited the *1/*3 combination, 7 (6.03%) exhibited *1/1*4, and one (0.86%) showed *4/*4. This research revealed significant differences in the occurrence frequencies of the CYP2D6 genotype between Han and Uygur ethnic groups, as well as between Uygur and Kazakh populations. However, no difference was found between Han and Kazakh populations. In addition, the prevalence of PMs of the Uygur is comparable to that of the Caucasians. However, the molecular mechanism underlying the poor metabolism is different in these two populations.
9.Investigation of the methylation status of Foxp3 gene during acute phase of Kawasaki disease
Guobing WANG ; Chengrong LI ; Jun YANG ; Pengqiang WEN ; Shilei JIA
Chinese Journal of Microbiology and Immunology 2010;30(7):678-682
Objective To investigate the methylation status of Foxp3 gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Thirty children with KD and eighteen agematched healthy children consented to participate in this study. Quantitative methylation specific polymerase chain reaction(MSQP) was used to assess the methylation status of Foxp3 promoter and regulatory T cells specific demethylated region(TSDR) in CD4+ T cells. The proportion of CD4+ CD25 + Foxp3 + regulatory T (Tr) cells was analyzed by flow cytometry. Transcriptional levels of CD4+ CD25 + Foxp3 + Tr associating genes (Foxp3, CTLA4, GITR, LAG3 and CCR8 ) and Foxp3-dependent molecules (UBD and LGAIS3)were measured by real-time PCR. Results ( 1 ) Demethylation level of Foxp3 promoter in CD4 + T cells from patients with KD was lower significantly than that of health subjects( P < 0.01 ), and increased significantly after treated with intravenous gamma globulin therapy(IVIG) (P < 0.01 ). No difference of demethylation level of TSDR region was observed among all groups ( P > 0. 05 ). ( 2 ) The proportion of CD4 + CD25 + Foxp3 + Tr in peripheral blood from patients with KD , as well as mRNA levels of Foxp3 gene in CD4+ T cells, was significantly lower than those of health subjects ( P <0. 01 ), and increases significantly after IVIG therapy (P <0.01 ). Significant positive correlations between demethylation level of Foxp3 promoter and the proportion of CD4 + CD25 + Foxp3 + Tr , or expression levels of Foxp3 in CD4 + T cells, were observed during acute phase of KD (CD4+CD25+ Foxp3+ Tr: r=0.76, P<0. 01; Foxp3: r=0.89, P<0. 01). (3)Transcription level of CD4+ CD25+ Foxp3+ Tr associating factors, such as CTLA4, GITR, LAG3 and CCR8, was significantly down-regulated in acute phase of KD(P<0. 01 ), and up-regulated to some extent after treated with IVIG(P <0. 01 ). Expression levels of Foxp3-dependent molecules UBD and LGALS3 in CD4 + T cells decreased significantly during acute phase of KD (P < 0.01 ), and basically recovered to the levels of health subjects( P < 0.01 ). Conclusion Decrease of demethylation level of Fopx3 promoter is correlated with immune dysfunction in Kawasaki disease.
10.Application of PBL Teaching Model in Medical Physics
Yuansheng LIU ; Jiqing YANG ; Xuemin QU ; Jun WEN
Chinese Journal of Medical Education Research 2003;0(02):-
PBL teaching model has been adopted as one of mainstream models in the west medical education.This article,with the characteristics of medical physics,combined with the advantage of PBL,discusses how PBL teaching mode is applied to medical physics.What we have done provides a reference for the development of PBL in medical physics.