We studied the function of connexin 43 (Cx43) gene in Xiao Meishan swine bone marrow mesenchymal stem cells (BMSCs) resulting from overexpression. Cx43 eukaryotic expression vector (pEGFP-Cx43) was constructed and transfected into BMSCs by nucleofector, after detecting the transfection efficiency; the expression of Cx43 was verified by RT-PCR, immunofluorescence and western blotting. Furthermore, we detected its cell cycle and apoptosis through flow cytometry. Our results show that pEGFP-Cx43 plasmid was successfully constructed, and green fluorescence in pEGFP-Cx43 transfected BMSCs was highly expressed with 60% transfection efficiency. In transgenic Xiao Meishan swines BMSCs, the expression level of Cx43 mRNA and protein were up-regulated. Meanwhile, the ability of cell proliferation was significantly increased, and the apoptosis rate was significantly reduced. Taken together, Cx43 overexpression could promote the proliferation of Xiao Meishan swine's BMSCs and markedly reduce their apoptosis, which provides evidence for in vivo research.
Animals ; Animals, Genetically Modified ; Apoptosis ; Cell Proliferation ; Connexin 43 ; metabolism ; Flow Cytometry ; Genetic Vectors ; Mesenchymal Stromal Cells ; metabolism ; Plasmids ; Swine ; genetics ; Transfection

Objective To explore the difference of DNA methylation levels between normal Schwann cells (NSCs) and activated Schwann cells (ASCs) in rats. Methods The adult Wistar rats were received sciatic nerve ligation and fed for 7 days. The ASCs and NSCs were separated from ligated sciatic nerves and brachial plexus respectively. Immunocytochemical staining of S-100 antibody was used to identify the cells. The growth condition of cells was detected by CCK-8 method. Methylated DNA immunoprecipitation sequencing (MeDIP-Seq) was applied to filter the differentially methylated regions in ASCs and NSCs. The distribution of differentially methylated genes related with axonal regeneration in chromosome was analyzed, and Gene ontology(GO)and PATHWAY analysis were also conducted. Results High purity of ASCs and NSCs were obtained successfully, which were both positive for S-100 antibody. In the same culture condition, ASCs showed a faster proliferation than that of NSCs. A total of 177 176 differentially methylated regions were found by MeDIP-Seq. Among them, 1 097 were located in the promoter (≤1 kb), 1 136 in the promoter (1-2 kb) and 567 on the CpG. After functional annotation of differentially methylated genes, 214 differentially methylated genes related with axonal regeneration were found in ASCs and NSCs. Compared with NSCs, 191 genes were up-regulated and 23 genes were down-regulated in ASCs. These genes were located on different chromosomes, most of which on chromosome 12 (22 genes) and the least on chromosomes M (2 genes). GO analysis indicated that the differential methylated genes were involved in axon growth, axon formation, axon elongation and axon guidance. The MAPK, cell adhesion molecules, Ras signaling pathway may be related with the differential methylated genes. Conclusion The methylation levels between ASCs and NSCs are significantly different, which are probably related with axon regeneration.

This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.
Alginates ; chemistry ; Barium ; chemistry ; Calcium ; chemistry ; Cartilage ; cytology ; Cations ; Cell Culture Techniques ; instrumentation ; Cells, Cultured ; Chondrocytes ; cytology ; Collagen Type II ; chemistry ; Glucuronic Acid ; chemistry ; Glycosaminoglycans ; chemistry ; Hexuronic Acids ; chemistry ; Metals ; chemistry ; Microscopy, Electron, Scanning

[Abstract ] In recent years, induced pluripotent stem cells (iPSCs) technology has played an important role in basic and clini-cal application research of Parkinson′s disease ( PD) and acquired significant progress .The neural progenitor /stem cells or dopamine ( DA) neurons which were obtained through iPSCs technique and direct differentiation technique from somatic cells were used for the study of cell therapy in PD , and good results were achieved .The cell models of DA neurons were established from PD patients carrying LRRK2, PAKK2, PINK or SNCA mutations via iPSCs technology , and the mitochondrial function and morphology , oxidative stress,α-synuclein ( SNCA) accumulation , and other aspects were studied on the pathogenesis of PD .This article briefly reviews the latest pro-gress of iPSCs technology in transplantation for treatment of PD and the establishment of cell model of PD disease , and provides refer-ence for further research .

Objective To confirm the immunomodulatory effect of ursolic acid on normal and immunosuppressive mice caused by cyclophosphamide.Methods We assayed the function of T lymphocyte transformation with MTT test,examined the function of B lymphocyte with hemolysis spectrophotography,determined the function of non-specific immunity with phagocytosis of macrophages,and measured the content changes of Th_(1) cytokine IL-2 and Th_(2) cytokine IL-6 in serum with ELISA.Results Ursolic acid had a suppressant effect on the function of humoral immunity and enhanced cell-mediated immune response at the low dosage but presented opposite effect at the high dosage in normal mice.Ursolic acid significantly improved immune function on immunosuppressive mice caused by cyclophosphamide(P

OBJECTIVETo investigate the impact of 1, 25-(OH)2D3 on left ventricular hypertrophy (LVH) in type 2 diabetic rats.

METHODSType 2 diabetic mellitus (DM) model rats were established by intraperitoneally injecting with 30 mg/kg streptozotocin. After 8 weeks, 19 male rats were identified as diabetic with left ventricular hypertrophy (LVH) by ultrasound examination, and randomly assigned into three groups: untreated (DM-LVH, n=7), treated with insulin (DM-LVH+INS, n=6), and treated with 1, 25-(OH)2D3 (DM-LVH+VD, n=6). Healthy male rats were used as the controls group (n=6). The fasting blood glucose and the insulin level were determined weekly. The left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor level were determined by 4 weeks later.

RESULTSIn the DM-LVH model group, the insulin level was significantly decreased compared with the non-diabetic control group (P<0.05), whereas the blood glucose, left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor expression were significantly increased (all P<0.05). In the DM-LVH+INS and DM-LVH+VD groups, the insulin levels were significantly increased compared with the DM-LVH model group (P<0.05), whereas the other parameters were significantly decreased (all P<0.05).

CONCLUSION1, 25-(OH)2D3 could reverse LVH in diabetic rats and that the mechanism may involve stimulating insulin secretion and reducing blood glucose via direct up-regulation of 1, 25-(OH)2D3-receptor expression.

Animals ; Blood Glucose ; analysis ; Calcitriol ; therapeutic use ; Diabetes Mellitus, Experimental ; blood ; complications ; Diabetes Mellitus, Type 2 ; blood ; complications ; Hypertrophy, Left Ventricular ; prevention & control ; Insulin ; blood ; Male ; Rats ; Rats, Wistar ; Receptors, Calcitriol ; analysis ; Streptozocin

The modern literatures on the diseases treated with acupuncture at Shenmen (HT 7) were collected. Using literature statistical analysis in bibliometrics, the common compatibility of Shenmen (HT 7) and the disease spectrum were analyzed so as to provide the references of the diseases that can be treated with acupuncture at Shenmen (HT 7). It was found that 39 kinds of diseases and symptoms, mainly nerve system disedses, could be treated with acupuncture at Shenmen (HT 7), including insomnia, depression, anxiety and dementia. Shenmen (HT 7) was seldom used alone. Instead, it was commonly used along with Baihui (GV 20), Sanyinjiao (SP 6), Neiguan (PC 6), Sishencong (EX-HN 1) and Taicong (LR 3).
Acupuncture Points ; Acupuncture Therapy ; statistics & numerical data ; Bibliometrics ; Humans

ObjectiveTo analyse the clinical characteristics of pancreatic intraductal papillary mucinous neoplasm (IPMN) which coexists with extrapancreatic malignancy (EPM),with an aim to provide strategies for clinical diagnosis and treatment.MethodsThe PubMed was used to search for the pancreatic IPMN related articles with positive pathologic results.A pooled analysis was then performed.The ratio ofpancreatic IPMNs coexisting with EPMs and the locations (or the type) of EPMs were analyzed.ResultsAfter a strict process of screening,18 articles met the pre-determined standardsand were accepted.Of the 1327 patients,363 had coexisting EPMs (27.35％).There were 392 EPMs in these 363 patients.The EPMs occurred in almost all the systems of the body,especially in the digestive tract and its related organs,which accounted for 63.06％ of the EPMs. Conclusions There is a tendency for patients with pancreatic IPMN to have coexisting EPM. More than half of these EPMs are malignant tumors in the digestive system. When pancreatic IPMN is diagnosed,the clinician should be aware of the possible coexistence of an EPM and should look for the possibility of a new EPM developing in a patient after treatment of pancreatic IPMN.

Objective:To describe the expression profiling of microRNAs of patients with non-small cell lung cancer. Methods:We reduced the scope of downregulated microRNAs in patients with non-small lung cancer by high throughput microarray based on quantitative real-time PCR (qRT-PCR). In this process, the downregulated microRNAs of non-small cell lung cancer tissues can be de-tected and compared with adjacent healthy lung tissues with relatively small samples. Different microRNA expression levels in non-small cell lung cancer and adjacent healthy lung tissues were verified in a large sample by RT-PCR. Results:A total of 20 types of microRNAs were significantly downregulated in non-small cell lung cancer tissues compared with adjacent healthy lung tissues. These microRNAs were listed as follows: hsa-miR-1; hsa-miR-22; hsa-miR-27a; hsa-miR-27b; hsa-miR-125a-5p; hsa-miR-125b;hsa-miR-142-5p;hsa-miR-143;hsa-miR-148a;hsa-miR-148b;hsa-miR-370;hsa-miR-373;hsa-miR-381;hsa-miR-489;hsa-miR-519a;hsa-miR-376c;hsa-miR-206;hsa-miR-380-5p;hsa-miR-223-5p;and hsa-miR-520c-3p. Among these microRNAs, 13 types were down-regulated in non-small cell lung cancer tissues as verified by RTPCR. These 13 types of microRNAs were listed as follows:hsa-miR-125a-5p; hsa-miR-143; hsa-miR-519a; hsa-miR-223-5p; hsa-miR-1; hsa-miR-520c-3p; hsa-miR-489; hsa-miR-27a;hsa-miR-373; hsa-miR-125b; hsa-miR-27b; hsa-miR-142-5p; and hsa-miR-206. Conclusion: In non-small lung cancer tissue, several kinds of microRNAs were significantly downregulated. Changes in microRNA expressions were significantly associated with tumori-genesis, progression, or other cancer cell functions in non-small cell lung cancer.