Objective To study the effect of allo-human bone marrow mesenchymal stem cells (BMSCs) on T and B cells from patients with Rheumatoid arthritis (RA) in vitro. Methods BMSCs were isolated from bone marrow samples of healthy volunteers and purified by density gradient centrifugation and cultured in vitro. Peripheral lymphocytes were isolated from patients with RA.Then, BMSCs and lymphpcutes were co-cultured. The modulatory effect of BMSCs on proliferation, activation and maturation of T and B lymphocytes of RA patients stimulated by PHA and SAC respectively was observed. The cell generation cycle and the degree of apoptosis were assessed by flow cytometry with PI/ Annexin V. After co-cultured with or without BMSCs for 72 hours, T cells were harvested, then they were labeled with anti-CD3, anti-CD4, anti-CD8, anti-CD25 antibodies and analyzed by flow cytometry. The density of IgG in the co-culture system was detected by ELISA. Results T and B cells proliferation was significantly suppressed when co-cuhured with bMSCs but did not induce T cell apoptosis. There was a significant decrease in the ratio of CD4+ CD3+ T cells in the co-cuhure group (34±6), as compared with the control group (44±7) (P<0.05). There was a decrease in CD25+ T cells and increase of CD4+ CD25+ cells in BMSCs co-cultured group (P<0.05). IgG was in creased in the cocuhure system. Conclusion Human BMSCs inhibit T and B cell activation and proliferation in patients with RA in vitro. And these immunomodulatory effects are not MHC restricted. The results of this study have provided evidence for the fact that BMSCs has the potential to be an effective treatment for RA.

Cancer-related fatigue is one of the most common and refractory clinical sympotoms in patients with cancer. Investigators have tried to study the molecular pathogenesis of cancer-related fatigue, and the cytokine has become a research focus. At present, a number of cytokines, including transformation growth factor, tumor necrosis factor, interleukin, proteolytic inducing factors, inflammatory cytokine are proved to be correlated to cancer-related fatigue.

Objective To study the effects of PIAS3 knocking down on the proliferation,cell cycle and apoptosis of human prostate cancer cell line DU145 in vitro.Methods PIAS3 specific short hairpin RNA(shRNA) expressing plasmid was constructed and named pSilencer4.1/PIAS3.DU145 cells were transfected with pSilencer4.1/PIAS3.The proliferation of DU145 cells was analyzed by MTT assay.Cell cycle and apoptosis of DU145 cells were analyzed by flowcytometry.Results PIAS3 shRNA expressing plasmid was succefully constructed and then confirmed by sequencing.Expression of PIAS3 in DU145 was significantly reduced after pSilencer4.1/PIAS3 transfection.MTT assay showned accelerated proliferation after PIAS3 knocking down,and showned dose-effect curve.Flowcytometry showed cells in S phase increased,cells in G0/G1 decreased and percentage of apoptotic cells decreased after PIAS3 knocking down.Conclusion Knocking down of PIAS3 expression accelerates DU145 cell proliferation and inhibit cell apoptosis in vitro.

Objective To explore the effects of recombinant expression plasmid of human glucagon-like peptide-1 (hGLP-1) analog gene (2?Val2-hGLP-1) on blood glucose, serum insulin level and pancreatic island in diabetic rats. Methods The diabetic rat model was reproduced by intraperitoneal injection of streptozotocin. The rats were then divided into 3 groups randomly (8 each): recombinant plasmid pIRES2-EGFP/Val2-hGLP-1 transfection group, empty plasmid pIRES2-EGFP transfection group, and diabetic rat model control group. Moreover, 8 untreated SD rats were set as normal control. Each rat in empty plasmid transfection group and recombinant plasmid transfection group was injected via tail vein with 110?g plasmid, whibe those in diabetic model control group and normal control group were treated with equal volume of normal saline solution. Blood glucose and serum insulin levels of rats were determined 30 days after experiment, and glucose tolerance test and insulin tolerance test were performed to estimate insulin sensitivity. The pathological changes in pancreatic island and insulin secretion were evaluated with HE and immunohistochemistry staining, respectively. Results Compare with normal group, diabetic model group and empty plasmid transfection group, the blood glucose level significantly lowered (P0.05). Meanwhile, the insulin secretion was increased and the pathological changes in pancreatic islands were alleviated in recombinant plasmid transfection group compared with that in diabetic model control group. Conclusions hGLP-1 analog gene transfection may be able to promote the proliferation of pancreatic islands and enhance sensitivity to insulin, thus significantly lower blood glucose level and ameliorate the lesion of pancreatic islets in diabetic rats.

[Abstract ] Objective The purpose of this study was to construct a short hairpin RNA (shRNA) interference lentiviral vector targeting the humanβ-COP gene and to evaluate its inhibitory effect on β-COP in THP-1 cells. Methods We designed and synthesized 4 humanβ-COP-specific oligonucleotide sequences and inserted them into the pGMLV-SC1 vector to construct a recombinant vector fol-lowed by transfection of HEK 293T cells with the recombinant vector and Lenti-HG Mix to produce lentiviruses and detect the viral con-tent.After infecting the THP-1 cells with the packaged lentiviruses , we analyzed the inhibitory effect of β-COP-shRNA on the β-COP gene by quantitative PCR and Western blot . Results Sequencing confirmed that the β-COP-specific oligonucleotide sequences were in-serted into the lentiviral vector and the lentiviruses were packaged in the transfected HEK 293T cells, with the final viral content of 1 × 109 TU/mL.Quantitative PCR showed that the 4 β-COP-shRNA vectors significantly decreased the mRNA expression of β-COP (P<0.01), with interference rates of 16.9 %,32.5%, 74.0%, and 50.3%, respectively.Western blot also indicated their inhibitory effect on the protein expression of β-COP, with an inhibition rate of 76.4% onβ-COP-shRNA3. Conclusion Lentiviral shRNA interference vectors targeting human β-COP were constructed successfully , which could suppress the expression of the human β-COP gene.

Objective To observe the efficacy of percutaneous vertebroplasty (PVP) combined with zoledronic acid in treatment of spinal metastatic carcinoma.Methods Fifty cases of spinal metastatic carcinoma were divided into control group and treatment group by random digits table method with 25 cases each.The control group was given zoledronic acid 4 mg,intervals of 28 d intravenous drip 1 time.The treatment group was given PVP besides zoledronic acid.The visual analog scale score,activity ability score and analgesic usage score after treatment for 3 months were observed and compared.The quality of life was evaluated and the occurrence of relapse of fracture was also recorded.Results After treatment for 3 months,the visual analog scale score,activity ability score and analgesic usage score were (2.00 ± 0.17),(2.12 ± 1.38),(1.47 ± 0.10) scores in treatment group,and (7.84 ± 0.18),(3.21 ± 0.13),(3.30 ± 0.09) scores in control group,and there were significant differences between two groups (P ＜ 0.01 or ＜ 0.05).The occurrence of relapse of fracture was 2 cases (8％,2/25) in treatment group and 9 cases (36％,9/25) in control group,and there was significant difference between two groups (P ＜ 0.05).Conclusions PVP combined with zoledronic acid in treatment of spinal metastatic carcinoma has many advantages such as small incision,short surgery time and rapid relief from pain.It can obviously improve the quality of life,and it is worthy of spreading in the clinic.

Objective: To better understand the antagonistic effect of Xiao Long Tong Bi (XLTB), a Chinese herb medicine, on α1-adrenoceptor (α1-AR). Methods: (1) Radio ligand binding assay . Specific　125I-BE2254(2-β(4-hdroxyphenyl)-ethyl amino-methyl-tetralone) binding was measured by incubating membrane of canine cerebral cortex with a single concentration of　125I-BE2254 in the presence of 15 concentrations of XLTB. Half-effectual concentration of inhibition (IC50) and Hill coefficients (nH) were determined by Hill plots. (2) Contractile responses of rat prostate strip in vitro were determined. pKB values for XLTB in competitively inhibiting NE-stimulated contraction of tissues were measured by the method of Ainlakshana. Results: XLTB competitively inhibited binding of　125I-BE2254 to α1-AR in a concentration -dependent manner. IC50 values for XLTB in canine cerebral cortex were (34.0±6.0)　g*L－1, the Hill efficiency value (0.7±0.1) was significantly decreased from unity. Contractile studies showed that XLTB competitively antagonized the NE concentration-response curve with pKB values of (37.0±11.0) g*L－1 or (30.0±8.0) g*L－1 when XLTB concentration was 70 g*L－1 or 170 g*L－1, respectively. The pKB values for XLTB in antagonizing NE-induced contraction of tissues were showed to fit in well with the IC50 values on rat prostate. Conclusion: These results suggest that XLTB appears to be a competitive antagonist for α1-AR.

BACKGROUND: Islet transplantation is an effective method for the treatment of type 1 diabetes mellitus and parts of type 2 diabetes mellitus. However, its application is hindered by insufficient sources and immunologic rejection. Though transdifferentiation of pancreatic stem cells is at the starting step, it is thought to be the hopeful source for islet cell transplantation.OBJECTIVE: To look for a suitable cells-transplantation source for the treatment of diabetes mellitus. METHODS: The pancreatic ductal epithelial cells were separated from Kunming mice and cultured in DMEM/F12 medium supplemented with keratinocyte growth factor, hepatocyte growth factor and nicotinamide, etc. Samples were taken at different time points for light microscopy and electron microscope. The changes of CK-19 and PDX-1 were detected by immunocytochemistry at 1 and 16 days. The expressions of insulin and glucagon gene were detected by RT-PCR at 1 and 16 days. The physiologic function of these islet-like clusters was determined by dithizone staining and glucose stimulation at 21 days. RESULTS AND CONCLUSION: A large number of epitheliod cells were CK-19 immunoreactive positive and few of them were PDX-1 positive at 1 day after isolation, then CK-19 positive cells proliferated quickly and formed substantial plaques of epithelial cells in cobblestone pattern. At 16 days later, these cells begin to form islet-like clusters gradually, while most of them were PDX-1 immunoreactive positive. The analysis of mRNA by RT-PCR showed very low levels of insulin and glucagon mRNA in the starting materials but increase was found as the process of transdifferentiation. At 21 day differentiated islet-like clusters were stained red by dithizone. In those samples exposed to a stimulatory 15 mmol/L glucose, there was a 1.6-fold increase in insulin compared with to 5.6 mmol/L glucose (P < 0.05). Pancreatic ductal cells of adult Kunming mice could proliferate quickly and have the potency of transdifferentiation into islet-like clusters when cultured in vitro under appropriate conditions.

Objective In order to choose the best surgical approach for minimally trauma varicocelectomy color Doppler ultrasound(CDU) was used to detect the anatomic relationships of spermatic vein in groin. Methods Sixty varicocele patients were randomly selected. Their spermatic veins were examined by CDU which beginning from superficial inguinal ring,passing the crossing point of spermatic vein and femoral artery ,and ending at the 3cm above the deep inguinal ring. The depths from skin to spennatic vein were measured and the relationships between spermatic vein and femoral artery were recorded. Results The average length of incision is 2.1cm and the average duration of operation is 22 minutes. The average depth from skin to spermatic vein was 1.1cm,1.55cm and 3.56cm respectively at the site of the superficial inguinal ring,the crossing point of spermatic vein and femoral artery,and the deep ingui-nal ring. Conclusion The best approach for minimally trauma varicocelectomy is at the crossing point of spermatic vein and femoral artery because here the spermatic vein is relative superficial and has merged into two or three vessels and the femoral artery can be easily touched by fingers.

Objective To evaluate the association between serum 25-hydroxyvitamin D3 [25 (OH) D3],parathyroid hormone,and arterial stiffness in patients with type 2 diabetes.Methods Serum 25 (OH) D3 and parathyroid hormone(PTH) were determined in a cross-sectional sample of 258 patients aged 30 years and over.Arterial stiffness was assessed by pulse wave velocity(PWV) obtained with a VP-1000 pulse wave unit.Fasting plasma HbA1c,lipid profile,calcium,and high sensitive-C reactive protein were determined.Results (1)The prevalence of vitamin D3 deficiency was high(79.84％) in patients with type 2 diabetes.(2) Those with lowered serum vitamin D3 levels had raised PWV [(1610.76 ± 142.70 vs 1527.95 ± 58.02) cm/s,P＜0.05].(3) Multiple stepwise regression analysis showed that 25 (OH) D3 was an impact factor of PWV risk score,which was independent of age,duration of diabetes,and systolic blood pressure(β =-0.256,P＜0.01).(4) Serum PTH was positively correlated with PWV (r =0.210,P ＜ 0.05) and systolic blood pressure (r =0.229,P ＜ 0.05),but negatively correlated with 25 (OH) D3 (r =-0.153,P ＜ 0.05).Conclusions 25 (OH) D3 deficiency is common in patients with type 2diabetes,and a low serum 25 (OH) D3 level is significantly associated with increased arterial stiffness in these patients.The association of serum PTH with arterial stiffness may result via changes in vitamin D and blood pressure.