1.Study on transplantation of induced bone marrow mesenchymal stem cells via various route for the treatment of chronic liver injury
Yan SUN ; Baorong CHI ; Lu CHEN ; Xiangwei MENG ; Dexia KONG
Chinese Journal of Digestion 2008;28(3):171-174
Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P<0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.
2.The influences of Decorin gene on biological behavior in human lung adenocarcinoma A549 cell line
Xue ZHANG ; Li WANG ; Xiangwei YAN ; Qingyong XU
Practical Oncology Journal 2016;30(4):289-294
Objective To explore the influences of decorin ( DCN) gene down-regulating on biological behavior in human lung adenocarcinoma A 549 cell line.Methods Chemically synthesis in vitro targeting DCN -siRNA was transfected into human lung adenocarcinoma A 549 cells by LipofectamineTM 2000 .The gene expres-sion of DCN was detected using Real-Time PCR ;The protein expression of DCN was investigated using Western blot;Checkout DCN-siRNA effects on lung adenocarcinoma cancer cell proliferation was detected by CCK -8;The migration and invasion ability were determined by Transwell assay .Results DCN-siRNA was successfully transfected into human lung adenocarcinoma A 549 cells.Real-Time PCR results showed that DCN mRNA ex-pression level significantly decreased ,compared with untransfected group ,negative control group .Western blot re-sults showed that DCN-siRNA transfection inhibited DCN protein expression level;CCK-8 results showed that the proliferation was enhanced in the DCN -siRNA group as compared with untransfected group ,as well as the negative control group and empty vector group .Transwell assay results showed the invasion of A 549 cells in DCN-siRNA group was significantly enhanced as compared with untransfected group [(22.6 ±1.14) vs.(5.2 ± 0.84)].Wound-Healing assay results revealed that the cell repairing rate was markedly increased in DCN -siRNA group.A549 cells were close to complete repair after 48 hours.Apoptosis was not significant in DCN -siR-NA group as compared with untransfected group (P=0.214).Conclusion Down-regulation the expression of DCN gene by DCN-siRNA may enhance the ability of invasion ,migration and proliferation of A 549 cells without affecting apoptosis ,which provides a new evidence of function for advancing research of lung cancer .
3.Prediction of the B cell epitope for MUC1 antigen
Qinghao YANG ; Xiangwei WANG ; Yan JIN ; Lixi ZHANG
Journal of Third Military Medical University 1988;0(05):-
Objective To predict the B cell epitope for MUC1 antigen. Methods The secondary structure and surface properties of MUC1, such as physical and chemical characters, hydrophilicity, antigenicity, were analyzed with various methods. Results Many distinct antigenic epitopes in MUC1 were identified by computation: 1-10, 24-54, 65-77, 84-91, 108-134, 140-156, 159-174, 179-196, 199-210, 220-233, 237-265, 270-299, 316-337, 351-362, 369-396, 411-420, and 445-502. Conclusion Prediction of the B cell epitope for MUC1 can provide a base for the studies of structure and function of MUC1, construction of its mutants, and selection of new expression forms of MUC1.
4.Effects of simulated microgravity on miRNA expression in mouse fibroblasts
Xiangwei SONG ; Zhanyu WANG ; Yufeng JIANG ; Bingxin XU ; Shaoyan SI ; Jinlian ZHOU ; Heming YANG ; Chenglin LI ; Yan CUI
Chinese Journal of Trauma 2017;33(8):756-764
Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of miRNA in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and divided into simulated microgravity (SMG) group and normal gravity (NG) group according to the simple random method.Samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled,and hybridized in sequence.Feature Extraction Software was used to collect the array images and get raw data,which were analyzed by Genespring Software.Differentially expressed miRNAs were identified and then validated by qRT-PCR.Target genes of differentially expressed miRNAs were predicted by the databases of Targetscan and microRNAorg,and the intersections of databases were identified as potential regulatory target genes.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were applied to determine the roles of these target genes.Relevant biological functions and/or signaling pathways of the regulated genes especially related with wound healing process were categorized based on their enrichments.Then integration predictions of the miRNA and mRNA expression profiles had been proposed to refine the functional miRNA-mRNA relationships.The miRNA-mRNA functional network and miRNA-mRNA control network were constructed.Results Four miRNA genes were up-regulated significantly including mmumiR-669j,-122-5p,-30a-3p,-6516-3p,among which mmu-miR-669j was up-regulated at 52.84 folds with the greatest significance (P < 0.05).Seventeen miRNA genes were down-regulated significantly including mmu-miR-21a-3p,-miR-28a-5p,-218-5p,-210-3p,-miR-19a-3p,-miR-31-3p,and-miR-19b-3p,among which mmu-miR-28a-5p was down-regulated at 15.47 folds with the greatest significance (P < 0.05).The qRT-PCR showed a high concordance with the microarray results (P < 0.05).Target gene prediction and functional enrichment analysis suggested that a variety of biological processes and signaling pathways involved in wound repair were significantly enriched (P < 0.05).Function network and regulation network of miRNA-mRNA covered all the differentially expressed miRNAs,which suggested that miR-21 a-3p and predicted target gene Smad3 might play an important role in wound healing under microgravity.Conclusions Simulated microgravity by RCCS can significantly affect the expression of stress-related miRNAs in mouse fibroblasts L929.The miRNA target gene prediction and functional enrichment analysis based on gene chip technology may provide theoretical basis for illustrating the mechanism and management of weightlessness stress injury.
5.Effects of sowing date on morphologic characteristics, yield and quality of Radix isatidis.
Yuhang CHEN ; Xiangwei YAN ; Qiaosheng GUO ; Hanqing TIAN ; Qiaohua DENG ; Yun PENG ; Jianying GUO ; Chengzhong GUAN
China Journal of Chinese Materia Medica 2009;34(21):2709-2712
OBJECTIVETo study the effects of sowing date on morphologic characteristics, yield and quality of Radix Isatidis, in order to provide a basis for selection of the suitable sowing date.
METHODThe experiment was carried out in a randomized complete block design arranged in split plot, the phonological period of Isatis indigotica was investigated and the morphological indexes, yield and quality at harvest were measured and determined by correlation and variance analysis.
RESULTThe different sowing dates showed a great effect on phonological period of I. indigotica. The biological characteristics, yield and active components of Radix Isatidis were negatively correlated with the sowing date, and the difference of morphological characteristics, yield and active compositions with sowing date was significant.
CONCLUSIONThe suitable sowing date of I. indigotica was during the period of time from late March to beginning of the April in Fuyang area, and considering the factors of rational rotation, the time cloud delayed until late May to the beginning of June.
Agriculture ; methods ; Biomass ; Isatis ; anatomy & histology ; growth & development ; Plant Roots ; anatomy & histology ; growth & development ; Seasons ; Time Factors
6.Contrast of growth characteristics of different germplasm and correlation between morphologic and individual yield of Isatis indigotica.
Qiaosheng GUO ; Yuhang CHEN ; Xiangwei YAN ; Hanqing TIAN ; Qiaohua DENG ; Jianying GUO ; Chengzong GUAN
China Journal of Chinese Materia Medica 2009;34(16):2034-2038
OBJECTIVETo study the growth characteristics of different germplasm and analyze the correlation between morphologic and individual yield of Isatis indigotica.
METHODThe planting samples were collected during the growth period, morphological characteristics and root yield per plant in 13 germplasm of I. indigotica from all over the country were determined by correlation and path analysis.
RESULTThe growth characteristics of various I. indigotica germplasm were consistent. The difference of morphological characteristics and root yield with various germplasm were significant. The root yield per plant was positively correlated to the main root length and the number of individual plant leaves.
CONCLUSIONThe number of leaves per plant and root length are selected for high-yield lines, meanwhile, leaf length and leaf diameter are important factors for breeding high-yield lines as well.
Biomass ; China ; Isatis ; anatomy & histology ; genetics ; growth & development ; Plant Leaves ; anatomy & histology ; genetics ; growth & development ; Plant Roots ; anatomy & histology ; genetics ; growth & development