AIM: To study the function of N-copine on cell level by investigating the effect of N-copine antisense on the cultured cortical neuron of mouse embryos. METHODS: Neuronal cultures were prepared from cerebral cortices of 16-day old mouse (BALB/C) embryos. N-copine expression was blocked by using antisense oligonucleotides. The effect of inhibition of N-copine expression on the cultured neuronal viability and development was observed. RESULTS: With oligonucleotides treatment for 72 h, the length of neuronal axon and the area of neurosoma in antisense group were decreased markedly as compared with those in control group. The number of trypan-blue-stained neurons and the LDH activity of the culture media were increased significantly as compared with those in control groups. CONCLUSIONS: Inhibition of N-copine expression affects the development of cortical neuron cultured in vitro, and induces the neuron demaged severely. The results suggest that N-copine is a functional protein in neurons, and it may play an important role in the regeneration of nervous system and preventing neuronal degeneration.

Objective To study the affect and the related molecular mechanism of glycogen synthase kinase-3β in the proliferation of osteosarcomaand its value in the target therapy of osteosarcoma.Methods The expression level of p-GSK-3β(Ser9)and GSK-3β were detected in human osteoblast cell and osteosarcoma cells by western blot.Observe the effect of GSK-3β inhibitors and siRNA interference on the GSK-3β regulate osteosarcoma cells using apoptosis protein chip.Evaluate the valueof GSK-3β target therapy on osteosarcoma in vivo.Results The expression level of p-GSK-3β (Ser9)was lower in osteosarcoma cells.LiCL,GSK inhibitor Ⅸ,siRNA knockdown could inhibit the cell viability and up-regulated the apoptosis-related protein cleaved-caspase3.The results of the protein array showed that downstream proteins of NF-κB downregulated significantly.The results were validated by western blot,while the downregulation of p-Iκ-Bα and nuclear NF-κB p65 were also observed after LiCL treatment.Inhibition of GSK-3β by either LiCl or specific siRNA resulted in a significant reduction of NF-κB luciferase reporter activity.Furthermore,the NF-κB luciferase reporter activity was significantly increased in CA cell lines,but not in KD cell lines.By contrast,NF-κB-luciferase reporter activity was significantly decreased in stably GSK-3β knockdown cells.GSK3β inhibitor LiCL and shRNA knock down demonstrated a strong cytotoxicity effect on osteosarcoma cells in vivo.Conclusion GSK-3β is in the state of relative active in osteosarcoma in osteosarcoma and important in cell proliferation.GSK-3β regulates cell survival partially through the NF-κB pathway.It is a promising therapeutic target in osteosarcoma.

Objective To investigate the expression level and clinical significance of melanoma antigen gene A (MAGEA) in osteosarcoma patients. Methods Compare gene expression profiles in osteosarcoma cell lines and osteoblasts with gene microarrays. Validation of differentially expressed genes was carried out by real-time polymerase chain reaction analysis. Corresponding protein levels were measures by Western blot analysis in osteosarcoma cell lines and by immunohistochemistry in osteosarcoma tissues. The staining intensity of immuno-histochemistry was correlated with clinical outcome , and its prognostic significance was analyzed. Results Sev-eral genes belonging to MAGEA increased significantly in all osteosarcoma cell lines and tumor tissue , but not in normal osteoblast cell. Patients with MAGEA expression has higher risk of lung metastasis (relative risk 2.79, 95% confidence interval, 1.12-6.93; P = 0.028) and lower five-year survival rates (39.6% ± 8.4% vs. 80% ± 8.9%, P = 0.01) compared with patients without MAGEA expression. Conclusions The expression of MAGEA increased in osteosarcoma , which inversely correlating with outcome of osteosarcoma patients.

Objective To explore the clinical efficacy of intensity-modulated radiotherapy combined with chemotherapy in the treatment of advanced cervical cancer.Methods All 121 patients with advanced cervical cancer (stage Ⅱ B &Ⅲ A & Ⅲ B) selected in our hospital from June 2012 to June 2014, who were treated with combined chemoradiotherapy, were divided into observation group [intensity modulated radiation therapy (IMRT)group)] and control group [3-dimensional conformal radiation therapy (3D-CRT) group].There was no significant difference between two groups in mean age, body mass index, International Federation of Gynecology and Obstetrics (FIGO) clinical stage, pathological type and chemotherapy mode (P ＞ 0.05).The clinical features of two groups were compared.The treatment and follow-up of two groups were recorded.Results The bone marrow suppression [32.2％ (19/59) vs 51.6％ (32/ 62)], gastrointestinal reaction [42.4％ (25/59) vs 62.9％ (39/62)], and rectal reaction rate [27.1％ (16/59) vs 45.2％ (28/62)] of the observation group were significantly less than that of the control group (P ＜ 0.05).The incidences of genitourinary tract injury [18.6％ (11/59) vs 24.2％ (15/62)], radiation proctitis [23.7％ (i4/59) vs 25.8％ (16/62)] and radiation cystitis [16.9％ (10/59) vs 20.9％ (13/ 62)] of two groups had no significant difference (P ＞ 0.05).Two groups were followed up for 3 years, the local control rate of the observation group was significantly higher than that of the control group (86.3％ vs 70.1％) (P ＜ 0.05).Conclusions IMRT combined with chemotherapy in the treatment of advanced cer vical cancer can improve the local control rate of tumor, protect the endangered organ, and reduce the side effects of radiotherapy.

OBJECTIVETo detect the expression of SCUBE3 in human osteosarcoma cell lines and surgical specimens of osteosarcomas and investigate its association with the patients' prognosis.

METHODSThe expression of SCUBE3 protein was detected in 5 osteosarcoma cell lines using Western blotting. CCK8 assay was used to assess the effect of SCUBE3 suppression mediated by two specific small interfering RNAs (siRNAs) on the proliferation of U2OS osteosarcoma cells, and the cell cycle changes were detected using flow cytometry. Immunohistochemistry was performed to detect the expression of SCUBE3 in 60 osteosarcoma tissues, and Kaplan-Meier method was performed for survival analysis of the patients.

RESULTSCompared with osteoblast hFOB1.19 cells, the osteosarcoma cell lines all showed significantly higher expressions of SCUBE3. In U2OS cells, suppression of SCUBE3 expression by siRNA significantly inhibited the cell proliferation (P<0.05). Kaplan-Meier survival analysis indicated that patients with high SCUBE3 expression had worse prognosis than those with low SCUBE3 expression (P=0.036).

CONCLUSIONSCUBE3 is up-regulated in the 5 osteosarcoma cell lines and in primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high SCUBE3 expression in osteosarcoma tissues is associated with a poor prognosis of the patients, suggesting that SCUBE3 can serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Calcium-Binding Proteins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Osteosarcoma ; metabolism ; pathology ; Prognosis ; RNA, Small Interfering ; Up-Regulation

Objective To investigate antimicrobial resistance among nosocomial gram-negative bacilli in 2006.Methods About 987 consecutive and non-repetitive gram-negative bacilli were isolated from 10 teaching hospitals from Sep.to Dec.in 2006 in China.All of these isolates were sent to the central laboratory for reidentification and susceptibility testing.The minimal inhibitory concentration(MICs)of meropenem and other antibacterial agents were determined by agar dilution method.Results The activity of antibacterial agents against Enterobacteriaceae was as fol lows in descending order of susceptible rate: meropenem(susceptible rate 99.8%),imipenem(99.5%),piperacillin/tazobactam(91.3%),amikacin (89.3%),cefepime(83.8%),cefoperazone/sulbactam(79.7%),ceftazidime(74.7%),cefotaxime (57.7%),ceftriaxone(56.6%),ciprofloxacin(53.6%).The prevalence of extended-spectrum β-Iactamases(ESBL)was 59.0% in Escherichia coli,33.0%in Klebsiella pneumoniae and 8.0%in Proteus mirabilis.The most active agents against E.coli and K.pneumoniae were meropenem,imipenem(99.2%. 100%),piperacillin/tazobactam(90.8%-97.0%),and amikacin(83.8%-92.4%).Cefepime Was more active against K.pneumoniae than E.coli(85.4% vs.65.2%).Against E.cloacae,E.aerogenes and Citrobacter freundii,the most active agents were as follows in desecnding order:meropenem,imipenem (99.2%-100%),amikacin(85.2%-92.6%),cefepime(81.5%-85.9%),piperacillin/tazobactam (73.4%-87.2%),cefoperazone/sutbactam(65.6%-77.7%),and ciprofloxacin(53.1%-72.3%).The most active agents against Pseudomonas aeruginosa were amikacin(83.5%),followed by meropenem (79.1%),piperacillin/tazobactam(74.1%),and imipenem(70.9%).The most susceptible agents against Acinetobacter baumannii were imipenem(79.1%),meropenem(73.4%) and cefoperazone/ sulbaetam(54.7%).Mutiresistant A.baumannii increased up to 53.0%.The most active agents against Burkholderia cepacia were meropenem(73.3%),eeflazidime(73.3%),and piperacillin/tazobactam (62.2%).Conclusions Carbapenems remained very high activity against Enterobacteriaceae.Increasing resistance to 10 antimicrobials agents tested from A.baumanni and P.aeruginosa brought great concern.

OBJECTIVETo detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.

METHODSRT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.

RESULTSSignificant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).

CONCLUSIONCXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokines, CXC ; metabolism ; Humans ; Osteosarcoma ; metabolism ; pathology ; Prognosis

80% activity rate against E.coli included piperacillin/tazobactam(93.4%)、ceftazidime(86%),and amikacin(83.3%);The susceptible rate to piperacillin/tazobactam in K.pneumoniae was 84.6%. The susceptible rate to ceftazidime decreased from 82.3% to 69.9%, which was lower than to cefepime (77.2%). Over 50% of Enterobacter cloacae were resistant to ceftazidime, cefotaxime and ceftriaxone. Susceptible rates to piperacillin/tazobactam in E. cloacae,E. aerogenes,Citrobacter freundii and Serratia marcescens (67.7%-96.4%) were higher than those to cefepime (68.8%-77.5%), cefoperazone/sulbactam (59.7%-87.5%). Susceptibility to amikacin among these 4 species (70%-83.7%) was higher than to ciprofloxacin (48.1%-79.5%). All of Morganella morganii and Proteus vulgaris isolates were susceptible to meropenem and imipenem; Over 90% of the isolates were susceptible to cefepime, cefoperazone/sulbactam and piperacillin/tazobactam.The most active agent against Pseudomonas aeruginosa was meropenem (84%), followed by amikacin, piperacillin/tazobactam, ceftazidime and imipenem (72.5%-76.6%). Mutiple-drug-resistant Acinetobacter baumannii increased from 33% in 2003 to 48% in 2004. Resistance to carbapenems increased to 18% in this species in 2004. The most active agents against Burkholderia cepacia were meropenme (64.9%), cefoperazon/sulbactam (63.2%), ceftazidime (59.6%), piperacillin/tazobactam (56.1%) and cefepime (52.6%).Conclusions Carbapenems remained very high activity against Enterobacteriaceae. Increasing resistance to 10 antimicrobials agents tested among A. baumanni brought great concern. Meropenem was 4-to 16-fold more active against common gram-negative bacilli than imipenem.