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.Churg-Strauss syndrome: report of a case.
Yan-wei QIAO ; Jun LIU ; Wen-jie YANG
Chinese Journal of Pathology 2012;41(7):488-489
6.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.
7.Study on biomarkers of silicosis and silica-exposed group.
Wen-jie JI ; Dong-jun CHENG ; Lei YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(3):217-218
Adult
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Aged
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Biomarkers
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blood
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Female
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Humans
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Laminin
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blood
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Male
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Middle Aged
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Occupational Exposure
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Risk Assessment
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Silicon Dioxide
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adverse effects
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Silicosis
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blood
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diagnosis
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Superoxide Dismutase
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blood
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Transforming Growth Factor beta
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blood
8.Development of single bus medical temperature measure system
Xuemin QU ; Yaling BAI ; Jun WEN ; Jiqing YANG ; Yuansheng LIU
Chinese Medical Equipment Journal 2004;0(08):-
This design is a multi-point thermometry system based on single bus digital thermometer DS1820.The single bus system of multi-point measure temperature is developed with single chip computer 89C52 and circuit units for physiological signal measurement.Obtainment method of higher resolution temperature data is given.The method makes measure temperature resolution reach to 0.1?C.It is characterized by simplicity of structure,high precision and real time,and is easy to transmit by internet network.So the system has widely applications value on clinic.
9.Design of pulsed magnetic fields stimulation instrument
Jun WEN ; Xuemin QU ; Jiqing YANG ; Sigang WANG ; Kaiping LONG
Chinese Medical Equipment Journal 2003;0(11):-
In this paper,a pulsed magnetic fields stimulation instrument is designed and realized,which provides a pulsed magnetic field with the range of maximal intensity from 0.01~2T,frequency from 0.2~100Hz and time width of pulse from 0.01~1ms.The instrument,controlled by the hand,foot or itself,can display stimulus intensity and times and output trigger signals with different waveforms to make measuring devices operate synchronistically.
10.Construction and identification of survivin-targeting siRNA-expressing plasmid
Xiaolin WANG ; Jianquan HOU ; Jun HE ; Shenmin YANG ; Duangai WEN
Chinese Journal of Clinical Laboratory Science 2006;0(01):-
Objective To construct survivin-targeting siRNA-expressing plasmid.Methods DNA sequence correspond to siRNA targeting survivin was designed and synthesized,and cloned into plasmid pRNAT-U6.1/Neo to produce surviving-targeting plasmid.Two oligos in the template with cohesive BamHⅠ and HindⅢ sites were prepared and annealled to form the insert fragment for siRNA vector.The vector was cut with BamHⅠ and HindⅢ and ligated with the insert fragment using T4 ligase.The recombinant vector was confirmed by restriction digestion and DNA sequencing,and then was transfected into T24 cells with Lipofectamine TM2000 and the expression of survivin was detected by real-time quantitive PCR.Results DNA sequencing for the PCR product showed that the recombinant vector pRNAT-U6.1/Neo-survivin was successfully constructed without any base pair mutation.The plasmid pRNAT-U6.1/Neo-survivin could efficiently reduce the expression of survivin and confer G-418 resistance in T24 cells.Conclusion The siRNA-expressing plasmid which were successfully constructed and transfected into T24 cells in this study may facilitate the application of RNA interference technique,and lay foundation for further studies on the function of survivin.