Elderly population is a group of people who need more medical care and acquiring immediate medi-cal treatment in time is important for the aged to get a good health status. The article demonstrates the differences of medical accessibility between rural and urban seniors and analyses the influencing factors and changes of the dispari-ties using the 2011 waves of CLHLS data. Results indicate that compared to rural seniors, the aged living in urban area are more likely to achieve immediate treatment when they are seriously ill. The mechanism of the disparities is made by the different socioeconomic development level and social and medical security system. Moreover, the main reasons not to visit doctor when necessary are having no money and inconvenience to travel;the proportion of having no money and far from hospital are significantly larger in rural area than urban.

AIM: To clone and express the metalloproteinase domain of human von Willebrand factor-cleaving protease (vWF-cp). METHODS: The metalloproteinase domain of human vWF-cp, amplified from the plasmid containing the vWF-cp cDNA gene by using polymerase chain reaction, was cloned into pUC18, and its accuracy was verified by sequencing. Then the domain was inserted into the multiclone site of pET28a(+) and included a 6?His Tag at its amino terminal. After induced by IPTG, the recombinant protein was purified by using a Ni-NTA column and confirmed by Western blot. RESULTS: Comparison of the nucleotide sequence of our cloned domain with the GenBank sequence revealed no difference. High-level expression of the recombinant protein was yielded after 5-hour induction, which amounted to 28% of total bacteria protein in inclusion body. Western blot demonstrated that it possessed high specificity. CONCLUSION: The metalloproteinase domain of vWF-cp was high efficiently expressed in Escherichia coli. This might contribute to the further study of the relationship between its structure and function. [

AIM: To investigate the therapeutic action of secreted endostatin (ES) on breast cancer cells. METHODS: Retroviral-mediated endostatin gene was transferred to breast cancer cell line MDA-MB-231. The ES biological properties and function were evaluated by polymerase chain reaction (PCR), MTT and a murine xenograft model. RESULTS: After retroviral transduction, endostatin genetically modified breast tumor cells were confirmed by PCR, and the integration and durative expression of endostatin gene was successfully committed. Compared with controls, endostatin secreted by genetically modified cells markedly inhibited endothelial cell proliferation (P0.05). The results of the transplanted subcutaneous tumor model in nude mice suggested that the subcutaneous growth of MDA-MB-231 was significantly inhibited by the expression of endostatin gene (P

Objective To investigate the effects of erythropoietin(EPO)on the function of glomerular endothelial cells in rats with chronic renal failure(CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats.Experimental rats were randomly divided into four groups:sham operation group (control group),CRF group,CRF rats treated with 30 U/kg EPO(low-dosage group)and with 50 U/kg EPO (high-dosage group).CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed.Serum creatinine(Scr),blood urea nitrogen fBUN),urine protein,haematoglobin (Hb) and blood pressure were measured.The renal morphologie changes were evaluated on periodic acid-schiff (PAS) stained sections.The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method.The mRNA of endothelin 1(ET-1),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus,and the expressions of eNOS and VEGF roRNA in renal tissue were higher in EPO treatment group than those in CRF model group(all P<0,05).The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group.In addition,the Scr,BUN,urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05).Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05).Reanl pathological injury wss improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure,maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

Objective To investigate the effects of erythropoietin (EPO) on the number and function of peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). Methods The model of chronic renal failure was established by a two-stage 5/6nephrectomy procedure in rats. Experimental rats were randomly divided into four groups (n =7,respectively): sham operation group, CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats were given EPO by hypodermic injection for 6 weeks, then EPCs were isolated by density gradient centrifugation from peripheral blood mononuclear cells. The ability of cell proliferation, adhesion and vasculogenesis in vitro was further observed. Results Compared to sham operation group, the ability of cell proliferation,adhesion and vasculogenesis in vitro in CRF rats was remarkably decreased (P＜0.05, respectively).Such ability was promoted significantly in dose-dependent manner by EPO treatment (P＜0.05,respectively). Conclusion EPO can improve the number and ability of endothelial progenitor cells from rats with chronic renal failure.

Objective To evaluate effects of acitretin and interferon?α(INF?α)alone or in combination on the proliferative activity of and interleukin?15 expression in human cutaneous T?cell lymphoma Hut78 cells. Methods Cultured Hut78 cells were divided into several groups, including blank control group, negative control group, dimethyl sulphoxide (DMSO) group and experimental groups. Cells in experimental groups were additionally classified into several subgroups to be treated with acitretin(0.1-10μmol/L, acitretin groups)or INF?α(5 000-20 000 IU/ml, INF?αgroups) alone, or the combination of 1.0 μmol/L acitretin and IFN?α at concentrations of 5 000- 20 000 IU/ml (combination groups), for 24, 48 and 72 hours. Subsequently, cell counting kit 8(CCK8)assay was performed to assess the proliferative activity of Hut78 cells, and enzyme?linked immunosorbent assay(ELISA)to measure the expression of IL?15 in these cells. Results The proliferative activity of and IL?15 expression in Hut78 cells were both obviously suppressed in the acitretin groups and combination groups compared with the DMSO group, as well as in the INF?αgroups compared with the negative control group, and the inhibitory effects gradually increased with the increase in acitretin or INF?αconcentrations and treatment durations. As repeated measures analysis of variance revealed, there was a significant difference in both proliferation inhibition rates and IL?15 expression among different treatment durations and among different concentrations of acitretin or INF?α(all P<0.05), and there was an interaction effect between treatment durations and drug concentrations(all P<0.05). A significant difference was observed in both proliferation inhibition rates and IL?15 expression at 24, 48 and 72 hours when the 1.0?μmol/L acitretin+ 10 000/20 000?IU/ml IFN?αgroup was compared with the 1.0?μmol/L acitretin group and 10 000/20 000 IU/ml IFN?αgroup(all P<0.05). There was also a significant difference in IL?15 expression at 24, 48 and 72 hours between the 1.0?μmol/L acitretin+50 000?IU/ml IFN?αgroup and 5 000?IU/ml IFN?αgroup(all P<0.05). Conclusions Acitretin and IFN?αboth can inhibit the proliferation of and IL?15 expression in Hut78 cells, the inhibitory effects are enhanced with the increase in drug concentrations and treatment durations, and the combination of acitretin and IFN?α appears to have stronger inhibitory effects than acitretin or IFN?αalone.

AIM: To investigate the protective effect of the bone marrow cells transfected with human multidrug resistance gene (MDR1) on the reconstruction of murine hematopoietic function.METHODS: The mononuclear cells of the bone marrow from donors, BALB/C mice, treated with 5-Fu previously, were isolated and transfected with human multidrug resistance gene in vitro , then transplanted to the tertiary recipients. After lethal irradiation(8.5 Gy) and bone marrow transplantation, the recipients were selected with Taxol 7 mg/kg intraperitoneal injection, VCR 5 mg/kg or DNA 5 mg/kg intravenous injection. The survival rate and blood pictures of mice as well as the integration and expression of target gene MDR1 were studied. RESULTS: The lethal irradiated murine hematopoietic function could be reconstructed and protected from toxicity of high doses Taxol, VCR and DNR selection after reinfusing the hematopoietic progenitor cells containing human multidrug resistance gene (MDR1). The survival rate and survival time of experimental mice were higher than that in the control group. The integration and expression of MDR1 gene in recipients were confirmed by PCR, RT-PCR and FCM. CONCLUSION: The integration and expression of human multidrug resistance gene in recipients may play an important role in the reconstruction and protection of murine hematopoietic function.

Objective To study the effect and safety of conversion from calcineurin inhibitors to rapamycin in kidney transplantation recipients with chronic allograft nephropathy.Methods In 82 kidney transplant recipients enrolled in this study,72 cases were diagnosed as having chronic allograft nephropathy by biopsy.Recipients (SRL group) were administered with rapamycin after withdrawal of calcineurin inhibitors.The doses of CNI in other recipients (non-SRL group) were not changed.Renal function,proteinuria,blood pressure,blood fat,hepatic function and hemogram were observed for 24 months in each group.Results During the follow-up period,serum creatinine level was dropped significantly in SRL group (P＜0.05),but it was increased in non-SRL group (P＜0.05).SRL group showed increased proteinuria,serum cholesterol and triglycerides (P＜0.05),and reduced Plt (P＜0.05).According to the renal function before conversion,the recipients who were administered rapamycin divided into four groups.In group A (Scr ＜ 120 μmol/L),there was no significant difference in diverse variables before and after conversion.In group B (Scr 120-200 μmol/L and Banff Ⅰ-Ⅱ),renal function was improved,and proteinuria alleviated.In group C (Scr 120-200 μmol/L and Banff ＞ Ⅱ),and group D (Scr ＞200 μmol/L),renal function was damaged to varying degrees and proteinuria was deteriorated.Conclusion It is safe and effective for patients with chronic allograft nephropathy to convert from calcineurin inhibitors to rapamycin.

Objective To investigate the effects of NOD2 on inflammatory responses of macrophages to Staphylococcus aureus. Methods Real-time RT-PCR detected NOD2 gene expression of macrophages infected by S. aureus. Synthesis of siRNA against NOD2 and interfere with macrophages, observed the effects of NOD2 gene silencing to phagocytosis of 5. aureus, cytokine secretion, activation of nuclear transcription factors, cell apoptosis of the macrophages infected by S. aureus using F.I .IS A, flow cytometry etc. Results S. aureus infection of macrophages can cause increased expression of intracellular NOD2. NOD2 gene silencing of macrophage lead to the decreased ability of phagocytosis with S. aureus, the lower levels of cytokines secretion, deficiencies of NF-κB activation. S. aureus can cause macrophage apoptosis, with the apoptosis rate increased with time. Conclusion The intracellular pattern recognition receptor NOD2 play a key role in pathogen recognition, signal transduction, activation of nuclear transcription factors in the process of macrophages infected by S. aureus.

Objective To investigate the expression of tumor necrosis factor alpha(TNF-α)and its relationship with infiltrating lymphocytes in lichen planus(LP).Methods Tissue specimens were obtained from the lesions of 60 patients with LP and normal skin of 20 human controls.Immunohistochemical SP method was used to detect the expression of TNF-α,and infiltrating lymphocytes were counted in TNF-α-positive tissue sections.Results TNF-α was expressed in 72％ of the LP specimens but in none of the control specimens(P ＜ 0.01).Positive staining for TNF-α was mainly located in the membrane of prickle cells,cytoplasm or membrane of dermal infiltrating lymphocytes.The expression of TNF-α in LP was uncorrelated with age,sex or disease course(all P ＞ 0.05),but was positively correlated with infiltrating lymphocyte number (rs =0.47,P ＜ 0.01).Conclusion TNF-α seems to play a certain role in the pathogenesis of LP.