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.

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 establish a method for content determination of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C in Wuzhi tablets.METHODS:RP-HPLC method was adopted.The determination was performed on Symmetry C18 column with acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set 225 rm,and the column temperature was 30 ℃.The sample size was 10 μL.RESULTS:The linear ranges of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C were 2.25-67.5ng(r=0.999 6),2.1-63 ng(r=0.999 8),28-840 ng(r=0.999 9),124.6-3 738 ng(r=0.999 9),22.7-681 ng(r=0.999 9),32.7-981 ng(r=0.999 9),47-1 410 ng(r=0.999 9),208-6 240 ng(r=0.999 9),5.36-160.8 rig(r=0.999 9),4.48-134.4 ng(r=0.999 8).The limits of quantitation were 14.17,13.32,9.33,11.37,14.62,19.88,14.66,12.50,16.40,13.55 rg.The limits of detection were 4.62,4.60,3.08,3.76,4.81,6.74,4.93,4.16,5.86,5.03 ng.RSDs of precision,stability and reproducibility tests were less than 3.0％;the recoveries were 96.36％-100.00％(RSD=1.83％,n=6),95.00％-100.00％(RSD=2.07％,n=6),95.00％-98.00％(RSD=1.22％,n=6),95.37％-98.91％ (RSD=1.29％,n=6),95.62 ％-103.71％ (RSD=2.85％,n=6),97.33％-102.67％ (RSD=2.00％,n=6),95.00％-99.33％ (RSD=1.75％,n=6),97.24％-104.93％ (RSD=2.63％,n=6),95.00％-97.50％ (RSD=1.42％,n=6),96.00％-102.00％ (RSD=2.45％,n=6),respectively.CONCLUSIONS:The developed method is accurate,sensitive and reproducible,and it can be used for content determination of 10 lignanoids in Wuzhi tablets.

Staphylococcus aureus (S. aureus) is an important human pathogen which can cause a chronic condition with a high relapse rate despite the aggressive antimicrobial treatment. Recent studies showed that intracellular pattern recognition receptors (including NOD) in response to bacteria or bacterial products play a proinflammatory role by activating nuclear transcription factor-κB (NF-κB). But how NOD2 mediates the proinflammatory response to S. aureus in mast cells (MCs) is unclear. So, in this study, we attempted to examine the role of NOD2 in inflammatory responses of MCs to S. aureus. P815 cells (a mouse mast cell line) were cultured. Real-time PCR was used to detect the NOD2 mRNA expression in P815 cells during S. aureus infection. The siRNA against NOD2 gene was synthesized and transfected into S. aureus-infected P815 cells. By using the methods of ELISA and flow cytometry, the effects of NOD2 gene silencing on cell phagocytosis, cytokine secretion, NF-κB activation and cell apoptosis of the S. aureus-infected P815 cells were examined. It was found that S. aureus infection could increase the expression of NOD2 mRNA in P815 cells. NOD2 gene interference in P815 cells reduced the number of S. aureus engulfed by P815 cells, the level of cytokines and the activation of NF-κB. In addition, S. aureus could induce the apoptosis of P815 cells, but NOD2 gene silencing did not affect the cell apoptosis rate. Our data suggested that NOD2 plays a key role in pathogen recognition, signal transduction, and NF-κB activation in the inflammatory responses of MCs infected by S. aureus.

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.

OBJECTIVE: To summarize the clinical experience of endoscopic repair for cerebrospinal fluid (CSF) rhinorrhea in our department in the last 4 years. METHOD: Clinical data of 16 patients with CSF rhinorrhea who underwent nasal endoscopic repair was analyzed retrospectively. The effect of etiology, image data, location of CSF leaks and surgical techniques on treatment were discussed. RESULT: Among the 16 patients, 10 were diagnosed with spontaneous CSF rhinorrhea, 2 were diagnosed with traumatic CSF rhinorrhea, 3 were diagnosed with CSF rhinorrhea after catching cold and 1 was diagnosed with meningo-encephalocele in with CSF rhinorrhea. The leak was located by CT scan in 11 cases, by MRI in 7 cases. The common locations of the defect were the frontal sinus (3 cases), cribriform roof (3 cases), sphenoid sinus (6 cases) and the nasal cavity top (4 cases). All the cases were successfully cured after the first nasal endoscopic repair with autologous materials. None of patients had a reoccurrence during 10 to 42 months follow-up time. CONCLUSION The application of CT and MRC before surgery which could make an accurate diagnosis of the location and the size of the defect. The correct selection of repair materials, processing planting bed around the leakage and complete contacting leakage with graft bed are the key points to the successful surgery of CSF rhinorrhea.
Adolescent ; Adult ; Cerebrospinal Fluid Rhinorrhea ; surgery ; Child ; Endoscopy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Failure ; Treatment Outcome ; Young Adult