Objective To investigate the expression of Polo-like kinase 1 in renal clear cell carcinoma and tissue adjacent to carcinoma;impact of BI2536 on the cell proliferation of renal clear cell carcinoma A498 cell line.Methods We select 12 cases of renal cancer Specimens of randical surgery between January 2013 and December 2014.The study included 7 male patients and 5 female patients,whose age ranged from 41 to 68 years(mean 49 years).Tumors were single and the location of tumor included 5 in left kidney,7 in right kidney.The size of renal tumor ranged from 2.1 to 6.3 cm.All patients did not have radiotherapy and chemotherapy preoperative who have no local and distant matastasis.The pathological results demonstrated renal clear cell carcinoma in 12 cases.At the same time,we collect the tissue 2 centimeters away as control,which pathological results demonstrated normal tissue.The expression of PLK1 was detected by western bloting in tissues from 12 cases of renal clear cell carcinoma and tissue adjacent to carcinoma and quantitative analysis was made.Cell proliferation of A498 cell line at the different concentrations of BI2536 were assayed by flow cytometry (0,20,40,60μg/L).Results The expression of PLK1 in renal clear cell carcinoma tissue are 0.74 ±0.2 and 0.21 ±0.13.The carcinoma tissue is higher than tissue adjacent to carcinoma (P =0.02).BI2536 act on A498 after 48 hours,the A498 cell number were arrested at G2/M phase at 0μg/L,20μg/L,40μg/L and 60μg/L were (10.28 ± 0.47) ％,(14.35 ± 0.85) ％,(20.49 ± 0.78) ％ and (23.90 ± 0.54) ％,respectively.The A498 cell number were arrested at G2/M phase increase when we incearse the concentrations of BI2536 at 48 hours (P ＜ 0.05).Conclusions The higher expression of PLK1 in renal clear cell carcinoma and the proliferation of the renal clear carcinoma cell can be repressed by BI2536 both reveal PLK1 may be used as the molecular maker and therapeutics target of renal clear cell carcinoma.

This paper is to evaluate the biocompatibility and cytotoxicity of a new Ni-free Zr-based bulk metallic glass (BMG), Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, by comparing it with conventional Ti6Al4V alloy. According to ISO 10993-5: 1999 and GB/T 16886.5-1997 standards, Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, pure Zr and Ti6Al4V materials were extracted with surface area of sample/volume of medium ratio being 1 cm2/mL and 0.5 cm2/mL, respectively. The viabilities of MG-63 cells (Human osteosarcoma cell line) cultured in the BMG medium extracts for 1, 3 and 5 days were determined by CCK-8 assay. The cellular morphology of MG-63 cells cultured on the surface of samples for 3 days was tested through laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM). The relative growth rate (RGR) of MG-63 cells cultured in Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and pure Zr were both more than 85%, indicating that the cytotoxicity of BMG was relatively low and met the national biomedical material eligibility standard. There was insignificant difference in the morphology of MG-63 cells cultured in the BMG medium extracts and the control group through LSCM and SEM, which showed the BMG had excellent biological compatibility. The Zr-based bulk metallic glass Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and the conventional Ti6Al4V alloy both had no obvious cytotoxicity to MG-63 cells. These results provided evidence that the new Zr-based bulk metallic glass could be potential replacement material for the orthopedic surgical implant.
Alloys ; Biocompatible Materials ; Cell Line ; Glass ; Humans ; Microscopy, Electron, Scanning ; Nickel ; Titanium ; Zirconium

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis

Objective To investigate the inhibitory effects of L. monocytogenes expressing melanomainhibiting activity(MIA)gene on malignant melanoma.Methods The plasmid pERL3-hly-MIA was constructed and used to transform live L.monocytogenes by electroporation.Atier culture.the transformed L.monocytogenes,namely LM-hly-MIA,was harvested for the detection of MIA protein by Westem Blot.A total of 24 mice were divided into three groups,namely control group,LM group and LM-hly-MIA group,to be immunized with physiological saline,L.monocytogenes and LM-hly-MIA,respectively.One day after the immunization.mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentration of 1×107/mL.One week later.reimmunization was performed.The size and weight of tumors were observed and measured in these mice.Results Enzyme digestion and Western blot confirmed the Successful construction of plasmid pERL3-hly-MIA.The average weight of tumors in mice was 4.33±0.91 gram.3.36 ±0.41 gram and 1.89±0.52 gram,respectively in the control group,LM group and LM-hly-MIA group,respectively(P＜0.05).with the inhibition rates of tumor growth being 22.4%and 61.5%in LM group and LM-hly-MIA group.respectively.Conclusion The attenuated strain of L.monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

Objective To investigate the association of promoter hypermethylation of secreted frizzled-related proteins (SFRPs) in patients with colorectal cancer. Methods The promoter hypermethylation of SFRPs in 20 sporadic colorectal cancer tissues and adjacent mucosa were detected by methylation-specific PCR. The amplified DNA was subcloned into the T-A cloning vector and sequenced. Two colorectal cancer cell lines (HCT116 and SW480) were treated with 5-aza-2' deoxycytidine for demethylation. The promoter hypermethylation and protein expression of SFRPs in colorectal cancer cell lines were detected by methylation-specific PCR and Western blotting. Results It was demonstrated that the hypermethylation of SFRP 1, 2, 4 or 5 was 19/20,17/20,3/20 or 13/20in cancer tissues, respectively, whereas it was 12/20, 8/12, 1/20 or 7/20 in adjacent mucosa,respectively. SFRP 1, 2 or 5 methylation was more frequently found in cancer tissue than in adjacent mucosa (P～0.05). Methylation of SFRP 1, 2, 4 and 5 were found in HCT116 cell line, but only SFRP1 and SFRP2 were found in SW480 cell line. There was a negative correlation between protein expression and methylation of SFRPs. The Western blotting revealed that SFRP protein re-expressedafter it treated with 5-aza-2' deoxyeytidine. Conclusion Methylation of SFRP 1, 2 or 5 gene is associated with the evolution of eolorectal cancer, and is closely related to silencing expression.

Objective To survey ST-segment resolution in STEMI patients undergoing emergency percutaneous coronary intervention (PCI) and to find the specific clinical features of patients with inadequate ST-segment resolution.Methods A total of 198 patients were divided into two groups according to the ratio of ST-segment resolution:relatively adequate ST-segment resolution group (＞ 50％) and inadequate STsegment resolution group (＜ 50％).The clinical features,infarct-related artery and PCI-related evants were evaluated,and major adverse cardiovascular events (MACE including target vessel revascularization,recurrent myocardial infarction,or death) were recorded during hospitalization and follow-up period.Multivariate logistic analysis was used to identify relevant factors influencing ST-segment resolution of STEMI patients after treatment with PCI.The Statistical analyses of data were carried out using SPSS 10.0 software.Results (1) There were 156 patients with relativey adequate ST-segment resolution and 42 patients with inadequate ST-segment resolution.Of them,there were higher percentage of patients aged over 75years in the inadequate ST-segment resolution group than those in the relatively adequate ST-segment resolution group (9 cases,21.4％ vs.14 cases,9.0％ ; P ＜0.05).(2) In inadequate ST-segment resolution group,thetotal ischemic time was significant longer [(5.2 ±2.2) h vs.(3.0 ± 1.6) h,P ＜0.01].The infarctrelated artery (IRA) was more common at left anterior descending coronary artery (LAD) (27 cases,64.3％ vs.69 cases,44.2％; P ＜ 0.05) and there were fewer patients with TIM grade 3 of IRA in inadequate ST-segment resolution group after primary PCI than that in relative adequate ST-segment resolution group (32 cases,76.2％ vs.140 cases,89.7％ ; P ＜ 0.05).There was a lower rate of using GP Ⅱ b/Ⅲ a receptor antagonist and a higher rate of prescribing IABP in inadequate ST-segment resolution group.(3) There is a higher incidence of MACE during hospitalization and follow-up period in patients with inadequate ST-segment resolution.(4) Multivariate logistic analysis indicated that age over 75 years,LAD occlusion,the total ischemic time were related to ST-segment resolution.Conclusions The patients with age over 75 years,LAD occlusion,longer ischemia time,and unemployment GP Ⅱ b/Ⅲ a receptor antagonist before PCI were prone to get inadequate ST-segment resolution and poor prognosis.Age over 75 years,LAD occlusion,and longer ischemic time were independent risk factors of the inadequate ST-segment resolution in STEMI patients after emergency PCI.

Objective To explore the development and interaction of left and right ventricular function in healthy children using tissue Doppler imaging.Methods Healthy children aged 0-15 years and adolescents were recruited,then children were divided into 6 groups:0-1 year,＞ 1-3 years,＞ 3-6 years,＞ 6-9 years,＞ 9-12 years,＞ 12-15 years.Healthy adolescents aged ＞ 15-25 years were also recruited.Every subject underwent echocardiography including cardiac dimension measurements,atrio-ventricular valvular velocity and early-diastolic flow velocity(E)/late-diastolic flow evlocity(A) ratio measured by pulsed color Doppler,atrio-ventricular annular myocardial velocity (including systolic velocity (s),early diastolic velocity (e) and late diastolic velocity (a)),time intervals (including isovolumic contraction time,ejection time and isovolumic relaxation time),isovolumic acceleration (ⅣA) and Tei index measured by tissue Doppler imaging.Results were compared among different groups,the correlations with age and other factors were explored.Furthermore,comparison was done between left and right ventricular functional parameters.Results Left ventricular Tei index and isovolumic contraction time were significantly lower during puberty.From infancy to pre-school stage,left ventricular E/A (flow velocity) and e/a(tissue velocity) increased accordingly,then presented with no significant changes among the following age groups(P ＞ 0.05).There were no significant differences in right ventricular Tei index,ⅣA,E/A (flow velocity) and e/a (tissue velocity) among the 6 groups (P ＞ 0.05).Left ventricular systolic myocardial velocity (s) and ⅣA were significantly lower than right ventricle (all P ＜ 0.001).However,left ventricular E/e(flow velocity) and e/a(tissue velocity) were significantly greater than right ventricle (all P ＜0.001).Conclusions In healthy children,left ventricular systolic function enhances during puberty,diastolic function increases from infancy to pre-school stage,then keeps stable till adolescents.Right ventricular systolic and diastolic function present with no significant changes during growth.Left ventricular diastolic function is greater than right one,however,right ventricular longitudinal systolic function is greater than left one.

ObjectiveTo investigate the effects of Taijiquan training on bone density and balance function of post-menopause women. Methods59 volunteers of post-menopausal women were divided into Taijiquan group and control group. Bone density was examined with DPX-L. The balance function were recorded with win-pop balance monitor and star diagram. ResultsBone density in lumbar vertebra, left collum femoris and left Ward's septa were higher in the Taijiquan group than in the control group. The lengths of center excursive loci reduced in Taijiquan group. The distances that the foot could touch were longer at Posterolateral, posteromedial, posterior, lateral(back) directions in the Taijiquan group than in the control group. ConclusionTaijiquan training can improve the bone density and balance function in post-menopause women.

Objective To investigate the inhibitory effect of live-attenuated Listeria monocytogenes (LM)-based vaccines expressing the gene encoding a melanoma differentiation antigen,MART-1,on malignant melanoma and their mechanism.Methods The constructed plasmid pERL3-MART-1 was used to transform live-attenuated LM by electroporation to construct recombinant LM.i.e.△inlB LM-MART-1 and △actA/△inlB LM-MART-1.The half lethal dose (LD50) of attenuated listeria strains was determined by concentration gradient dilution method.C57BL/6 mice were randomly divided into three groups,namely PBS group,△inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group.Mice were inoculated by intraperitoneal injection of O.1 LD50 of each rLM strain or PBS only.One week later,the mice were injected subcutaneously with 1×105 B16F10 cells(a mouse melanoma cell strain)in 200μl of PBS.Reimmunization was performed on day 14 and 21.Subsequently,the growth of tumor and survival of tumor bearing mice were observed.All mice were killed on day 28,and tumor tissue as well as splenocytes were obtained from these mice for the detection of MART-1 gene expression by real-time quantitative PCR and the percentage of CD4+CD25+T cell by flow cytometry.Results The recombinant △inlB LM-MART-1 and △actA/△inlB LM-MART-1 were constructed successfully.The LD50 of △inlB LM and △actA/△inlB LM was lower than LM-EGDe by 100 and 10 000 times respectively.Compared with PBS,the tumor growth was inhibited with △inlB LM-MART-1 by 46.95%(F=6.3,P<0.05),and by 83.96% with △actA/△inlB LM-MART-1(F=37.8,P<0.01).The relative expression level of MART-1 in △inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group was 8.988±0.207 and 11.315±0.445 times that in PBS group (both P<0.05).The percentage of CD4+CD25+T cells in splenocytes was (2.52±0.20)%,(1.14±0.13)% and (0.44±0.15)% in PBS group,△ialB LM-MART-1 group and △actA/△inlB LM-MART-1 group,respectively;the differences were statistically significant between the three groups (all P

Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.