1.Construction and application in renal cell carcinoma 769-P cells of miR-218 expression plasmids
Long HE ; Yan ZHANG ; Jun WANG ; Long LIU ; Lianhui FAN
Cancer Research and Clinic 2013;25(12):806-808,811
Objective To construct a stable expression plasmids miR-218,to lay the experimental foundation for the expression and mechanism of miR-218 in human renal cell carcinoma.Methods The primers were designed by Has-miR-218 and the miR-218 fragment were amplified by PCR,and then which was connected with the carrier pcDNA3.1 (+) to build a stable expression plasmids.The recombinant plasmids were trasfected into renal cell carcinoma cell line 769-P,and the expression of miR-218 in these cells were detected.Results The plasmids were constructat successfully,which was determind by enzyme digestion and sequencing results.The constructed expression plasmids pcDNA3.1-miR-218 transfection of human renal cell carcinoma cell line 769-P,miR-218 cxpression level of cells was significantly increased after trasfected by recombinant plasmids carrying miR-218 gene (relative expression quantity was 1.64).Conclusion The miR-218 expression plasmids pcDNA3.1-miR-218 were successfully constructed,the plasmids can be applied to the study of miR-218 function and mechanism in renal cell carcinoma.
2.Advance on stereotactic radiotherapy for early stage non-small cell lung cancer
Yan-Yang WANG ; Xiao-Long FU ;
China Oncology 2000;0(06):-
Stereotactic radiotherapy is a rapidly growing treatment strategy for tumor due to the advances in target definition,radiation dose delivery,and image guidance of radiation.We reviewed the clinical application of stereotactic radiotherapy in the management of early stage non-small cell lung cancer including the technique,the workflow of the technique established,the outcome and relative radiation toxicity.
3.The Development of Endogenous CO and its Mechanism in Asthma
Hong WANG ; Yan LONG ; Jing WANG ; Weili LUAN
Chinese Pediatric Emergency Medicine 2001;8(1):20-21
Objective To explore the development of endogenous CO and its mechanisms in asthma.Methods 50 guinea pigs were equally separated 5 groups.After the models of asthmatic guinea pigs were established in 4 groups,3 groups of them were treated with dexamethasone,the specific stimulator(hemin)and inhibitor(Sn-PP)of HO-1 respectively.The rest of groups were asthmatic one and normal control one(NCG).From all tested animals,the COHb in blood and lung HO-1 activity were examined,the lung tissues were pathologically observed and immunohistochemically stained.Results In asthmatic and hemin stimulated groups,the levels of COHb with enhanced expression of lung HO-1 and their activity were significantly increased compared with NCG(t=4.43~9.97,P<0.01).For example,the lung HO-1 activities in asthmatic group were (881±361)pmol/(mg*pro*h),COHb (4.94±2.15)% respectively.In dexamethason prevented and Sn-PP inhibited groups,however,all determined levels were markedly reduced compared with asthmatic group(P<0.01).Conclusion The increased HO-1 had resulted in the growth of endogenous CO in asthma.
4.Study on characteristics of pharmacological effects of traditional Chinese medicines distributing along kidney meridian based on medicinal property combination.
Ying-Long REN ; Hao GU ; Yun WANG ; Yan-Jiang QIAO
China Journal of Chinese Materia Medica 2014;39(13):2413-2416
To study the characteristics of pharmacological effects of property combinations of traditional Chinese medicines (TCMs) distributing in the stomach meridian based on medicinal property combination, in order to further define the association relationship between properties of TCMs and their pharmacological effects, and build a bridge for the interpenetration and combination between the medicinal property theory of TCMs and their pharmacological effects. On the basis of the studies on the medicinal property theory of TCMs distributing along the kidney meridian and their pharmacological effects, efforts were made to collect relevant data for medicinal properties and pharmacological effects and mine the characteristics of pharmacological effects that were corresponding to relevant medicinal property combination by processing materials related to medicinal properties and pharmacological effects with the association rules method. According to the analysis, TCMs distributing along the kidney meridian with different medicinal property combinations were significantly differentiated in the pharmacological effects, but shared identical pharmacological effects, such as immunological enhancement. In this study, TCMs distributing along the kidney meridian with different medicinal property combinations were taken as the carriers to closely integrate the traditional Chinese medicine theory with the modem study achievements, lay a solid foundation for further developing and enriching the traditional Chinese medical property theory, while providing a new perspective on the development of modem medicine.
Databases, Factual
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Drugs, Chinese Herbal
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chemistry
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therapeutic use
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Humans
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Kidney
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drug effects
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Kidney Diseases
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drug therapy
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Meridians
6.Preliminary study of the biological characteristics of fibroblasts in human granulation tissue in vitro.
Yan LONG ; Dali WANG ; Zairong WEI ; Changmin GUO
Chinese Journal of Plastic Surgery 2014;30(3):203-209
OBJECTIVETo explore the biological characteristics of fibroblasts in adult granulation tissue in vitro, including cell viability, CD phenotypic, factor and protein expression, and differentiation, so as to facilitate further research of the role of fibroblasts in early wound healing.
METHODSFibroblasts were isolated from human granulation tissue and cultured by mechanical and enzymatic digestion method. The cell morphology and proliferation were observed under inverted phase contrast microscope. The first and third passages of cells' growth curve were drawn respectively. The surface markers (CD105, CD73, CD90, CD44, CD34, CD45, CD19, CD11b, HLA-DR) of the first and third passage fibroblasts were identified by Flow Cytometry. And the expression of Vimentin, CK19, CD31 and Factor VIII were detected by immunocytochemistry.
RESULTSPrimary cultured fibroblasts were short spindle, polygonal and irregular in shape. The morphology of fibroblasts were uniform by repeatedly passage cultured in vitro and showed spindle-shaped. The proliferative capacity of the fibroblasts were not significantly different, with logarithmic growth phase. From 1 to 4 days, The primary and third passage cells' proliferation was no difference (P > 0.05), After the five days, the proliferative ability of third generation was better than the primary passage (P < 0.01). All fibroblasts highly expressed mesenchymal stem cells' surface markers CD105, CD73, CD90 and CD44, and didnt express hematopoietic stem cells' surface markers CD34, CD45, CD19, CD11b and HLA-DR. The expression of mesenchymal stem cells' surface markers in third generation of cells were increased significantly. Immunocytochemistry showed positive expression of Vimentin, CD31, and negative expression of CK19 and Factor VIII.
CONCLUSIONSFibroblasts in human granulation tissue show the biology characteristics of Mesenchymal Stem Cells. Some biological markers of endothelial cells are expressed in fibroblasts in granulation tissue. The fibroblasts may play an important role during the process of endothelial to mesenchymal transformation in early wound healing.
Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; Gene Expression ; Humans
8.Significance of histopathologic analysis vs cytomegalovirus (CMV) detection in the diagnosis of CMV-infected hepatitis post liver transplantation
Long CHENG ; Shuguang WANG ; Zhanyu YANG ; Ke ZHANG ; Xiaochu YAN
Chinese Journal of Organ Transplantation 2009;30(6):348-350
Objective To evaluate the sensitivity and specificity of histopathologie analysis vs cytomegalovirus (CMV) detection for the diagnosis of CMV-infected hepatitis post liver transplantation. Methods Twenty-five biopsies with CMV infection and twenty-five without CMV infection were collected. Histopathologic observation, immunohistochemical staining and virus detection were performed on both groups to evaluate the sensitivity and specificity of these examinations for the diagnosis of CMV-infected hepatitis. Results The detection rate of microabscess, aggregation of monocyte and rnacrophage, and cytomegalic change in CMV infection group was higher than that in the group without CMV infection (P<0.05), but there was no significant difference in intranuclear inclusion and eosinophilic body between the two groups (P>0.05). The sensitivity and specificity of IHC and PCR for CMV detection were 20% and 100%, 72% and 84%, respectively. Conclosions CMV detection with PCR combined with histological observation is the most effective diagnostic scheme for CMV disease of liver.
9.Verification of colon-targeted release of Changankang Pellets
Huijun YAN ; Zhixian LONG ; Yurong WANG ; Li ZHEN ; Weihua GAO
Chinese Traditional Patent Medicine 1992;0(07):-
Objective:To determine the in vitro release of two kinds of Changankang Pellets (Radix Astragall, Rhizama coptid) coated with Surelease and Eudragit respectively and verify their colon-targeted release in vivo through X-ray photography. Methods: Changankang Pellets were made by mixing the medical BaSO 4 powder and appropriate drugs together according to original produce and coat methods. After determining their in vitro release, they were tested in vivo by 2 volunteers. X-ray films were taken after appropriate time and analyzed to determine the location and status of these pellets inside healthy objects. Results: Pellets made by this way can carry drugs through stomach, intestine and targeted-release them in colon; the coating combined with pH-dependent and time-dependent Eudragit thin layer has better results. Conclusion: X-ray radiography applying BaSO 4 powder could be taken as a quality control method for colon-targeted release drugs. The preliminary results showed that Changankang Pellets met the requirements of colon-targeted release.
10.Construction and expression of small interfering RNA expression vector of CXCR4
Yan LIU ; Long WANG ; Dawei ZHANG ; Deming ZHAO
Journal of Jilin University(Medicine Edition) 2006;0(06):-
Objective To construct the recombinant plasmid carrying small interfering RNA(siRNA) to CXCR4 and observe dumbness effect of chemokine receptor gene CXCR4 suppression by RNA interference(RNAi),and explore the new method of more efficacious therapeutic possibilities for HIV-1.Methods Small interfering RNAs targeting CXCR4 gene were designed by using software.The complement form was obtained by annealing and interted into vector Psilencer3.1-H1neo.After sequencing,MT4 cells were transfected using the plasmid vector.RT-PCR and flow cytometry detection were used to evaluate the suppression of CXCX4 expression in different groups.Results The recombinant plasmid had the correct special fragments and DNA sequence detected by PCR,electrophoresis and DNA sequencing;The siRNA obviously suppressed the expression of CXCR4.When transfected with plasmid vector for 48 h,the inhibitory rate was 70%,compared with control and negative groups,there were significant differences(P