OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.

ObjectiveTo study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro.MethodsESC was induced decasualization in vitro and matched with non-decidualized cells as controls.The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR.Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it.Results (1) Between decidualized and undecidualized stromal cells,there were 49 miRNAs significantly different expression by microarray chip,including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b,30c,143,101,181 b,29b,30d,507,23 a,222,221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P ＜0.05).(2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC,ESC were cultured by FBS medium for 24 hours,the rate of transfection was 70％.ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours,the percentage of ESC at Sphase of (6.2 ± 0.7 ) ％ were significantly lower than ( 10.9 ± 0.8 ) ％ in control group ( P ＜ 0.05 ) ; the percentage of ESC at G0/G1 phase increased at transfection group [ (77.5 ± 1.3 ) ％ vs.(73.0 ± 1.6) ％ at control group ],but there was no significant difference (P ＞ 0.05 ).Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h,the percentage of ESC at S-phase was ( 3.3 ± 0.6) ％ in transfection group,which were significantly lower than (7.8 ± 0.9 ) ％ in control group ( P ＜ 0.05 ).The percentage of ESC at G0/G1 phase was ( 80.7 ± 1.6 ) ％ in transfection group and ( 74.9 ± 1.1 ) ％.In control group,which did not reached statistical difference ( P ＞ 0.05).ConclusionmiRNA was involved in ESC decidual process in vitro by regulating cell cycle.

Objective A preliminary study was conducted on the measuring tool for teachers' attitude in the medical schools towards curriculum integration of medical program.Methods Literature review and expert interview were conducted to design a questionnaire which comprised 56 items to investigate teachers' attitude towards medical curriculum integration.The method of factor analysis and Pearson correlation coefficient were used to evaluate structure validity,the content validity index I-CVI and S-CVI were used to evaluate the content validity and Cronbach's alpha coefficient and split-half reliability were used to evaluate the reliability of the questionnaire.Results Three principal components were extracted by factor analysis,which explained 59.219％ of the total variance.After rotation,the factors 1-3 were respectively defined as value recognition,participation initiative and emotional tendency.Cronbach's alpha coefficient of the three factors and all the items is 0.922,0.903,0.728 and 0.862 respectively.Conclusion The reliability and validity of the measurement tool is acceptable,and can be used as a reference for similar domestic survey

Background and purpose: Hepatocellular carcinoma (HCC) is a hypervascular tumor associated with a poor prognosis and lack of effective treatments. Consequently, identifying novel therapeutic strategies are urgently needed. We have previously shown that the kringle 1 domain of human hepatocyte growth factor (HGFK1) is a more effective anti-angiogenesis molecule than angiostatin. In this study, we observed the effects and mechanisms of HGFK1 gene on the HCC. Methods: A recombinant adeno-associated vires carrying the HGFK1 gene (rAAV-HGFK1) was constructed.HCC of rat was induced by McA-RH7777. rAAV-HGFK1 was used to treat the rat, median survival time and metastasis rate were observed. Results: Ten days after tumor cell inoculation, surgery were performed to confirm the tumor formation, PBS, rAAV-EGFP or rAAV-HGFK1 was injected directly into the tumor nodule followed by portal vein injection. Results from our study demonstrated that rAAV-HGFK1 treatment significantly prolonged the median survival time of the HCC bearing rats from 30 days (PBS and rAAV-EGFP groups) to 49 days (rAAV-HGFK1 group). More importantly rAAV-HGFK1 inhibited tumor growth and completely prevented liver, lung and peritoneal metastasis. In the controlled PBS and AAV-EGFP group, liver and peritoneal metastasis rate were both 100%, and lung metastasis rate was 100% and 83%, respectively. While there was no metastasis found in treatment group, with only 33% of ascites happened. This was most possibly due to the primary tumor in liver but not due to the metastasis. Moreover, at a higher magnification (1000×), it was clear that the HGFK1 protein was expressed mainly in the cytoplasma of liver cells. In parallel, IHC staining of CD31 also demonstrated a significantly lower level of microvessel density (MVD) (6.21±1.6) in the liver tumor of the AAV-HGFK1 treatment group, as compared to the two control PBS and AAV-EGFP groups (25.1±2.1 and 26.8±2.5, respectively, P<0.01). HE staining showed that AAV-HGFK1 treatment induced large areas of necrosis in the tumor tissues, while minimal areas of necrosis were observed in the tumor tissue in the control groups. In addition, no toxicity appeared when high dosage (4.8× 1012 vg/rat) of rAAV-HGFK1 was administered in rats. Conclusion: Results from this study demonstrated that HGFK1 inhibited the growth and metastasis of HCC and prolonged the survival time of animals with HCC through anti-angiogenesis effects. No obvious toxicity was observed. It might be the novel promising treatment for HCC and other cancers.

Objective To evaluate the value of echo‐contrast RT‐3DE for assessment of left ventricular volume and function in patients with left ventricular non‐compaction(LVNC) .Methods Twenty‐one patients of LVNC were involved and underwent non‐enhanced and contrast‐enhanced RT‐3DE to evaluate left ventricular end‐diastolic volume (LVEDV) ,left ventricular end‐systolic volume (LVESV) ,left ventricular ejection fraction (LVEF) .The endocardial border definition of LV was graded for each of the 16 LV segments as follows :0 = border invisible ,1 = border visualized only partially ,and 2 = complete visualization of the border .Three image‐quality groups (good ,fair ,and uninterpretable) were identified . Results ①Duringcontrast‐enhancedRT‐3DE,ascomparedwithnon‐enhancedRT‐3DE,thenumberof segments with complete visualization of the endocardial border increased significantly (55% vs 82% ,P <0.01) ,and the number of patients with a good‐quality echocardiogram increased significantly (33% vs 81% , P <0.01) .②Contrast‐enhanced RT‐3DE provided significantly larger values of LVEDV ( P < 0 0.1) and LVESV ( P < 0 0.1) as compared with non‐enhanced RT‐3DE ,the values of LVEF were not statistically different between the two techniques ( P =0.07) .③Intra‐and inter‐observer agreement for assessment of LV volumes and systolic function improved during contrast‐enhanced RT‐3DE ,as compared with non‐enhanced RT‐3DE .Conclusions Contrast‐enhanced RT‐3DE can increase the prevalence of good‐quality echocardiograms and significantly improve the reproducibility of LV volumes and function measurements .

Objective To investigate the effect of COX￣2 inhibitor celecoxib on radiosensitity of irradiation￣resistant cell line CNE￣2R of nasopharyngeal carcinoma and the potential mechanism. Methods Via exposing to a series of X￣ray (2, 4, 6, 8 Gy, 3 times for each dose), radio￣resistant cell subline CNE￣2R was established from human nasopharyngeal carcinoma cell CNE￣2.Radiosensitivity was detected by clone formation assay.CNE￣2R and CNE￣2 cell lines were exposed to 25, 50, 75 μmol.L-1 celecoxib, respectively.Western blotting was used to detect the protein expression of COX￣2.Clone formation assay was performed to measure the survival fraction of CNE￣2 and CNE￣2R after radiotherapy alone or radiotherapy combined with 30 μmol.L-1 celecoxib treatment.Flow cytometry was used to measure influence of radiotherapy alone or radiotherapy combined with 30 μmol.L￣1celecoxib treatment on cell apoptosis.Number of residual γ￣H2AX foci was observed by immunofluorescence assay. Results The colony forming assay demonstrated that the values of SF2, D0 , Dq , and N of CNE￣2R cell subline [(0.81±0.05), (2.15±0.07) Gy, (2.94±0.08) Gy, (3.91±0.07), respectively] was significant higher than those of CNE￣2 cell line [(0.61±0.08), (1.47±0.06) Gy, (1. 68 ± 0. 10) Gy, (3. 13 ± 0. 05), respectively]. The expression of COX￣2 protein was significantly downregulated with increasing celecoxib concentration.Surviving fraction was decreased in both CNE￣2 and CNE￣2R cell lines after irradiation.After radiotherapy combined with celecoxib, apoptosis rates of CNE￣2 and CNE￣2R cell lines [(13.10±0.63)%, (5.30±0.75)%] were higher than those of the corresponding control groups [(4.90±0.71)%, (1.82±0.82)%].Celecoxib increased radiosensitivity in nasopharyngeal carcinoma CNE￣2R and CNE￣2 cell lines.The number of residual γ￣H2AX foci after irradiation was increased by celecoxib pretreatment.The difference was statistically significant (P<0.05). Conclusion Celecoxib can enhance radiosensitivity of radio￣resistant cell subline CNE￣2R of human nasopharyngeal carcinoma in vitro.

In this study, 5'-CMP was sulfonated, and then the modified 5'-CMP was connected to a protein carrier as an immunogen to immunize BALB/c mice. After cell hybridization, screening and recloning , a McAb (B10) with high sensitivity and specificity was selected. In a dot Hot using the McAb B10, less than 0.05 pg of sulfonated DNA could be detected while 10 ng of DNA was not coloured The result showed that the sensitivity of McAb B10 was higher than that of the McAb from "Chemiprobe" kit

OBJECTIVE: To evaluate the anxiety and depression status of patients with nasal septum deviation before and after surgery, and explore the relationship between anxiety and depression and nasal symptoms. METHOD: By means of visual analogue scale (VAS) as the evaluation index of quality of life, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were used to analyze statuses before and after surgery. The 98 patients with nasal septum deviation were collected. RESULT: The preoperative SAS/SDS scores of the nasal septum deviation patients were higher than those of the national standards. The level of anxiety and depression were lower than the national norm after treatment. The patients of the normal and moderate SAS/SDS scores felt better after surgery. The patients of the severe SAS/SDS scores felt the symptoms improve poor. CONCLUSION Doctors should pay attention to the application of the SAS/SDS in patients of nasal septum deviation. Surgeon should actively communicate with severe patients in psychological problems, and strictly control the surgical indication of such patients.
Anxiety ; complications ; Depression ; complications ; Humans ; Nasal Septum ; abnormalities ; Psychiatric Status Rating Scales ; Quality of Life ; Self Report

We reported a case of gastric perforation repair operation performed by the first domestic surgical robot, named Miaoshou. The comprehensive and accurate nursing assessments and targeted psychological care were performed before the surgery. Close observation and care, effective prevention and treatment of complications, and nice guidance of diet were conducted after the surgery. All of these measures exerted positive effects on successful operation and early rehabilitation for patients.
Humans ; Perioperative Nursing ; Robotic Surgical Procedures ; Stomach Diseases ; nursing ; surgery

Objective To develop an evaluation index system of reputation for higher medical insti-tutions and to test its reliability and validity. Methods Using literature method and small-group discussion to collect the evaluation indexes, the data was collected by the questionnaire survey and the 1 493 effective questionnaires were collected with effective recovery rates of 61.90%. And the exploratory and confirmatory factor analysis methods were conducted to develop evaluation index system with using the SPSS 17.0 soft-ware for statistics analysis. Results The evaluation index system of reputation for higher medical institutions included six dimensionalities: social responsibility, opinion direction, social image, management quality, emotional attract and environmental facility. The Cronbachs’a coefficients of evaluation index system were greater than 0.7, RMSEA is 0.063<0.08, χ2/df is 3.933<5, therefore, the structural equation model fitting was good and evaluation index system had good validity. Conclusion This article has developed an evalu-ation index system of reputation for higher medical institutions, which has a good reliability and validity. To test the model’s reliability and validity, the suggestion that the future research will be performed between more other medical universities has been given.