AIM:To study the effects of transforming growth factor-β_1 (TGF-β_1) on cell apoptosis,cell cycle,production of endogenous TGF-β_1,expressions of P27~(Kip1),cyclin E and bcl-2 mRNA levels in NB4 cells. METHODS:Apoptotic morphological changes were observed by Wright-Giemsa staining. Cell cycle and apoptosis were detected with flow cytometry. Semiquantitative RT-PCR was used to examine the mRNA levels of endogenous TGF-β_1,P27~(Kip1),cyclin E and bcl-2. RESULTS:TGF-β_1 significantly restrained the growth and promoted the apoptosis of NB4 cells. The blockage of NB4 cells treated by TGF-β_1 at concentration of 5 μg/L was in G1 phase. Endogenous TGF-β_1 mRNA expression in NB4 cells was up-regulated when the concentration of exogenous TGF-β_1 was <5 μg/L. Meanwhile,the expression of endogenous TGF-β_1 mRNA was down-regulated when the concentration of exogenous TGF-β_1 was 10 μg/L. After treated with TGF-β_1 at concentration of 5 μg/L,P27~(Kip1) mRNA expression in NB4 cells was up-regulated,cyclin E and bcl-2 were reduced. CONCLUSION:TGF-β_1 is able to induce apoptosis and cell cycle distribution abnormally in NB4 cells by (1) Up-regulation of endogenous TGF-β_1,so that NB4 cells was induced into apoptosis through consequently high expression of P27~(Kip1). (2) TGF-β_1 may lead to cell cycle arrest by inhibiting the expression of cyclin E directly,or by inhibiting the activity of cyclin E through the increased expression of P27~(Kip1). (3) Down-regulation of bcl-2 induces apoptosis of NB4 cells.

Objective To observe insulin resistance (IR) in non-diabetic peritoneal dialysis (PD) patients,and analyze its related factors.Methods The non-diabetic PD patients who had been on stable PD at least three months were eligible to enroll.The patients were measured for their height,weight,waist to hip ratio,fasting glucose,fasting insulin,lipids and other biochemical indicators,dialysis adequacy indicators in August 2012,and divided into two groups depended on median HOMA-IR in August 2012.Results A total of 56 patients were enrolled and divided into two groups according to median HOMA-IR,including high IR group (HOMA-IR≥ 1.79,n=29) and low IR group (HOMA-IR ＜ 1.79,n=27).Compared to low IR group,high IR group were older [(57.9±14.2) years vs (48.7±14.5) years],had higher daily dialysate glucose load [(138.7±28.5) mmol/L vs (114.0± 21.5) mmol/L],higher waist-to-hip ratio [(0.91±0.08) vs (0.86±0.07)],higher BMI [(23.0±3.0) kg/m2 vs (21.2±3.1) kg/m2],higher triglycerides [(2.51±1.36) mmol/L vs (1.42±0.48) mmol/L],lower high-density lipoprotein cholesterol [(1.00±0.27) mmol/L vs (1.23±0.32) mmol/L],and lower Kt/V [(1.74±0.37) vs (2.08±0.56)].Multivariate logistic regression showed that age (β=0.122,P=0.033),triglycerides (β=1.798,P=0.030) and daily dialysate glucose load (β=0.094,P=0.031) associated with the degree of insulin resistance.Conclusion More dialysate glucose exposure is a risk factor of the occurrence of insulin resistance in non-diabetic patients with peritoneal dialysis.

After communication with professional course teachers, clinical experts and graduates by investigation, informal discussion and expert interviews, the course and teaching of rehabilitation therapy was reformed to make the students not only meet the skill requirement,but also acquire basic theory and sustainable develop in their career.

OBJECTIVETo study the change of WT1 gene expression during human leukemic K562 cell differentiation and apoptosis induced by bufalin.

METHODSCell viability was determined by trypan blue exclusion, cell differentiation and apoptosis by nitro blue tetrazolium (NBT) reduction test, morphology and flow cytometry, expression of WT1 protein by Western blot analysis, and expression of WT1 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTS(1) The cell proliferation was inhibited by bufalin and the IC(50) at 24, 48, 72 h were 0.026, 0.032 and 0.006 micro mol/L, respectively. (2) Bufalin induced K562 cell differentiation towards macrophage/monocyte within concentration from 0.01 to 0.05 micro mol/L and apoptosis at higher than 0.026 micro mol/L. (3) The expression of WT1 protein and mRNA were downregulated by bufalin in the initial stage of differentiation and apoptosis induced by bufalin.

CONCLUSIONK562 cell differentiation and apoptosis induced by bufalin might relate to the downregulation of WT1 expression.

Apoptosis ; Cell Differentiation ; Cell Proliferation ; Down-Regulation ; Humans ; WT1 Proteins ; genetics

OBJECTIVE: To establish an in vitro method to culture mesenchymal stem cells(MSCs) derived from human nasal mucosa, and explore their stemness and differentiation potential. METHOD: Based on the observation of distribution of MSCs in human nasal mucosa, we cultured and proliferated MSCs in vitro and identified the expression of stem cell markers on them including Nestin, CD133, Vimentin and Sa114 with immunofluorescence. The MSCs were induced to differentiate to osteoblasts with medium containing dexamethasone, ascorbic acid and beta sodium glycerophosphate, and to neurons with Neurobasal medium containing B27, ATRA and TSA. Histochemistry and immunofluorescence were applied to evaluate the differentiation. RESULT: The nestin and vimentin immunofluorescence-positive MSCs existed extensively in human nasal mucosa. While the MSCs were cultured in the osteogenic-inducing medium, activities of alkaline phosphatase were increased significantly, and bone nodules were found on the surface of the osteoblasts by alizarin red staining. After the induction by neural-inducing medium, the MSCs adopted neuron like appearance with many slim protrusions interconnected as a network. The induced cells expressed neural markers NF-200 and BM88 strongly. CONCLUSION The MSCs derived from human nasal mucosa are multipotent stem cells and can be utilized as seed cells to repair bone or neural injury.
Alkaline Phosphatase ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Humans ; Mesenchymal Stem Cells ; cytology ; metabolism ; Multipotent Stem Cells ; Nasal Mucosa ; cytology ; Neurons ; Osteoblasts ; cytology