Objective To construct and identify recombinant adenovirus vectors expressing GFP and human vascular endothelial growth factor (VEGF) iRNA and to inhibit the gene expression of VEGF in human nasopharyngeal carcinoma cell line CNE with the RNA interference (RNAi) technique,and explore the expression of VEGF in CNE and its proliferation before and after transfection.Methods By the method of restriction endonuclease digestion,the GFP and human VEGF genes were subcloned from pGenesil-1 plasmid into the shuttle vector pGSadeno-GFP-VEGF.After correct identification of the recombinant plasmid pGSadeno-GFP-VEGF,it was then digested by Pacl and transfected into HEK293 cells to be packaged and amplification for rAd-VEGF.Meanwhile,the recombinant adenovirus was identified by PCR method.The expression of GFP was detected to evaluate the titre and transfective ratio.Results Digestion and PCR identification showed the construction of recombinant adenovirus rAd-VEGF was successful via LR recombinant.The titre was 2×109 PFU/ml while the transfective ratio in CNE cell line was more than 90 percent which met the need of experiment.Cell growth was inhibited compared with non-siRNA transfected CNE cell line.Conclusion The recombinant virus expressing GFP and VEGF are successfully constructed; it layed a foundation for further relevant study.

0.05)indicating no significant loss of correction.The length of the growth of instrumented spine was 13.3 mm.No sever complications in the series. Conclusion The PRSS which dispenses without bony fusion is a safe and an effective instrumen for management of juvenile scoliosis.It provides and maintains desirable scoliosis correction in one stage procedure,while allowing spinal growth.

Objective To investigate the regulatory effects of PTEN-PI3K/AKT pathway on the apoptosis of gastric cancer MKN28 cells and the possible mechanisms.Methods The specific sequence of PTEN was transfected to MKN28 cells by eukaryotic expression vector (transfection group),and then vacant vector (negative control group) and PBS (blank control group) were transfected to the MKN28 cells,respectively.The effects of PTEN-PI3K/AKT pathway on the apoptosis of MKN28 cells and the expressions of PI3K,AKT,Caspase-3 and Caspase-9 were investigated.The growth curve and apoptosis of the MKN28 cells were detected by MTT assay and TUNEL staining,respectively,and the protein expression was detected by the Western blot.MKN28 cells which did not transfected by the PTEN were processed by inhibitor of PI3K (LY294002) (treated group),and MKN28 cells in the control group were processed by PBS.The expressions of apoptosis protein and apoptosis-related protein were detected after inhibition of PI3 K.All data were analyzed using the t test.Results The model of over-expression of PTEN was obtained and transfected into MKN28 cells.The survival of MKN28 cells in the transfection group was significantly inhibited in a time-dependant manner ( r =0.938,P ＜ 0.05 ).The mean apoptotic rate of the transfection rate was 27.86％ ± 4.78％,which was significantly higher than 0.01％ ± 0.01％ of the negative control group ( t =9.527,P ＜ 0.05 ).The protein expression of PI3K in the transfection group was 0.25 ± 0.03,which was significantly lower than 0.93 ± 0.16 of the blank control group and 0.96 ± 0.15 of the negative control group (t =7.235,8.883,P ＜ 0.05 ).The protein expression of P-AKT in the transfection group was 0.21 ± 0.03,which was significantly lower than 0.93 ± 0.13 of the blank control group and 0.91 ± 0.12 of the negative control group (t =9.347,9.802,P ＜ 0.05 ).The expressions of Caspase-3 and Caspase-9 of the transfection group were 0.86 ± 0.11 and 0.87 ± 0.12,which were significantly higher than 0.16 ± 0.03 and 0.18 ± 0.04 of the negative control group,and 0.15 ± 0.02 and 0.16 ± 0.03 of the blank control group ( t =10.634,10.999,9.448,9.942,P ＜0.05).The apoptotic rate of the MKN28 cells of the treated group was 28.60％ ± 4.50％,which was significantly higher than 0.12％ ± 0.06％ of the control group (t =10.961,P ＜ 0.05 ).The protein expression of PI3K and P-AKT of the treated group were 0.18 ± 0.02 and 0.11 ± 0.01,which were significantly lower than 0.93 ± 0.14 and 0.90 ± 0.12 of the control group (t =9.186,11.363,P ＜ 0.05 ).The protein expression of PTEN of the treated group was 1.15 ± 0.15,which was significantly higher than 0.21 ± 0.08 of the control group (t =9.577,P ＜ 0.05 ).The relative expressions of Caspase-3 and Caspase-9 of the treated group were 0.86 ± 0.12 and 0.88 ± 0.11,which were significantly higher than 0.25 ± 0.02 and 0.21 ± 0.03 of the control group (t =8.685,10.178,P ＜ 0.05).Conclusions Over-expression of PTEN may enhance the apoptosis of gastric cancer cells through inhibition of PI3K/AKT pathway.Inhibition of PI3K can enhance the expression of PTEN.PI3K and PTEN regulate the apoptosis of gastric cancer cells.

Objective To explore the correlation between wedged hepatic vein pressure (WHVP) and directly measured portal vein pressure (PVP) and further analyze the correlation between hepatic venous pressure gradient (HVPG) and portal pressure gradient (PPG).Methods From December 2012 to April 2014,the related data including WHVP,free hepatic venous pressure (FHVP),inferior vena cava pressure (IVCP) and PVP of patients who received transjugular interahepatic portosystem stentshunt (TIPS) treatment were collected,and HVPG and PPG were calculated.The correlations between WHVP and PVP,between FHVP and IVCP,between HVPG and PPG were analyzed.Pearson's correlation analysis were performed for correlation analysis.Results Twenty two patients matched the criteria were enrolled during the December 2012 to April 2014.The mean pressures of PVP and WHVP were (28.07±4.43) mmHg (1 mmHg=0.133 kPa) and (26.22±5.91) mmHg,respectively.PVPand WHVP were positively correlated,the correlation coefficient of them was 0.431 (P=0.045) and slope was0.323.The mean pressures of FHVP and IVCP were (7.31±3.37) mmHg and (6.82±4.01) mmHg,respectively.FHVP and IVCP were positively correlated,the correlation coefficient of them was 0.845 (P＜0.01) and slope was 0.711.The mean pressures of PPG and HVPG was (21.02±3.76) mmHg and (18.90±4.86) mmHg,respectively.There was no correlation between PPG and HVPG,the correlation coefficient of them was 0.014 (P=0.951).Conclusions There is a good correlation between PVP and WHVP,and so is the correlation between FHVP and IVCP.However,there is no good correlation between HVPG and PPG in this study because of the effects of many factors.

Objective To investigate the correlation between hepatic venous pressure gradient (HVPG) and clinic features ,laboratory results in patients with liver cirrhosis .Methods From December 2012 to April 2014 ,patients with liver cirrhosis who received HVPG examination were enrolled .The clinical data of the patients were collected ,which included etiology of cirrhosis ,albumin ,creatine ,total bilirubin ,international normal ratio (INR) ,history of ascite and bleeding ,degree of gastroesophageal varices under endoscopy ,the scores of Child‐Pugh and model for end‐stage liver disease (MELD) .Single factor and multiple factor linear regression method were performed to analyze the correlation between these indexes and HVPG .Results A total of 63 patients met the inclusion criteria .Among them ,six patients had abnormal shunt in liver venous and HVPG examination failed .The HVPG of the left 57 patients was 9 .50 to 33 .20 mmHg (1 mmHg = 0 .133 kPa) ,mean (16 .38 ± 5 .64) mmHg .The results of single factor regression analysis indicated that there were certain relevance between the level of albumin (r2 = 0 .145 , P= 0 .002) ,Child‐Pugh score (r2 = 0 .069 ,P= 0 .048) and HVPG .Multiple factor analysis indicated that there were certain relevance between albumin (B= - 4 .920 ,t= - 3 .521 ,P= 0 .001) ,total bilirubin (B =4 .066 ,t= 2 .206 ,P = 0 .032) and HVPG ,and there were no relevance between the other indexes and HVPG .Conclusion Only albumin and total bilirubin level in patients with liver cirrhosis are correlated with the level of HVPG .

Objective:To explore the influence of ulinastatin combined with thymosinα1 on the immune function of patients with a-cute brain injury. Methods:Sixty-eight cases of patients with acute brain injury were divided into the control group and the observation group randomly with thirty-four ones in each. The control group was given the routine treatment, and the observation group was given ulinastatin combined with thymosinα1 additionally. After the 1-day, 3-day, 7-day and 14-day treatment, transforming growth factor-β1 (TGF-β1), interleukin-6(IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) and the other serum inflammatory cyto-kine levels, and CD4 +, CD8 +, CD4 + /CD8 +, HLA-DR and cellular immune index levels were detected in the two groups. The prog-nosis effects were evaluated by the prognostic classification of brain injury, and the adverse reactions were analyzed in the two groups as well. Results:After the 1-day treatment, there were no significant differences in the serum inflammatory cytokines and immune param-eters between the groups (P>0. 05). After the 3-day, 7-day and 14-day treatment, serum TGF-β1, IL-6, IL-10 and TNF- α levels were higher than those on the first day after the treatment, and TGF-β1 showed an increasing trend with time extension, while IL-6, IL-10, TNF-α and CD4 + and CD4 + /CD8 +rose first and then decreased. After the 3-day, 7-day and 14-day treatment, serum IL-10 and CD4 +levels in the observation group were significantly higher than those in the control group, and IL-6 and TNF-αlevels were signifi-cantly lower than those in the control group (P<0. 05). After the 3-day and 14-day treatment, CD4 + and CD8 + levels in the observa-tion were significantly higher than those in the control group, and after the 7-day and 14-day treatment, HLA-DR levels were signifi-cantly higher than those in the control group (P<0. 05). The prognosis effect of the observation group was better than that of the con-trol group with statistically significant difference (P<0. 05). Conclusion:Ulinastatin combined with thymosin α1 is used to treat the patients with acute brain injury with better cellular immune function improvement and prognosis effect, which is worthy of clinical popu-larization and application.

Objective To explore the effectiveness of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients with EGFR double mutations containing rare mutations.Methods From March 2007 to January 2017,NSCLC patients with EGFR double mutations containing rare mutations confirmed by histopathology and EGFR mutation detections at Renmin Hospital of Wuhan University,Zhongnan Hospital of Wuhan University and Hubei Cancer Hospital were enrolled.According to the mutation types,patients were divided into double activating mutations group and activating mutations with insensitive mutations group.The effectiveness of TKIs in NSCLC patients with EGFR double mutations containing rare mutations and different mutation types was analysis.Results Among the 2 637 NSCLC patients underwent EGFR mutation detections,19 patients were confirmed as EGFR double mutations containing rare mutations.Fifteen patients received EGFR-TKIs therapy,the objective response rate (ORR),disease control rate (DCR) and median progression free survival (PFS) were 46.7％ (7/15),73.3％ (11/15) and 8.1 months,respectively.In patients with double activating mutations,two patients had partial response (PR),one patient had a stable disease (SD).In the activating mutations with insensitive mutations group,five patients had PR,three patients had SD,four patients had no effect.Conclusion Patients with EGFR double mutations containing rare mutations have a general response to EGFR-TKIs,and this result can provide a reference for the treatment of those patients.

Objective To study the risk factors for pathological upgrading after diagnosis of esophageal low?grade intra?epithelial neoplasia with ESD preoperative biopsy. Methods The endoscopic and pathological data of 85 lesions with ESD preoperative biopsy were analyzed, and grouped based on pathological upgrading after ESD. The risk factors for pathological upgrading after ESD was studied through single and multiple factor analysis. Results Pathological upgrading occurred in 45(52?94%) lesions after ESD, among whom 38 lesions developed up to high?grade intra?epithelial neoplasia and 7 lesions developed to esophageal early cancer. NBI?ME was performed on 37 patients and the accuracy of detecting the pathological invasion was 83?8%(31/37).Multi?factor analysis showed that reddish surface(OR=9?478, 95%CI:2?775?32?368, P = 0?000 3 ) and nodular lesion ( OR = 15?628, 95%CI:1?475?165?617, P =0?022 5) were independent factors for pathological upgrading after ESD. Conclusion Pathological upgrading of low?grade intra?epithelial neoplasia was common, especially esophageal mucosa with red surface and nodular lesion.Biopsy combined with NBI?ME is of significant importance to improve diagnostic accuracy.

For the last decade,the incidence of kidney neoplasms has shown an obvious rising trend in the world. The most common histopathological type of kidney neoplasms is clear cell renal cell carcinoma (ccRCC),which has a poor prognosis. ccRCC is generally accompanied by reprogramming of glucose,fatty acid,glutamine,tryptophan and arginine metabolic networks and pathways. Reprogramming of metabolic net-works and pathways enables tumor cells to proliferate rapidly,survive in conditions of nutrient depletion and hy-poxia,and escape surveillance by epidemic systems. New strategies have been developed to the treatment of ccRCC by targeting key proteins or enzymes involved in metabolic reprogramming pathways.

Objective To investigate the effects of microRNA-134 (miR-134) on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) and its potential molecular mechanism.Methods Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the differences of miR-134 expression between 10 cases of lung cancer tissues and normal lung tissues,and between normal human lung epithelial cell line BEAS-2B and lung adenocarcinoma cell line A549.miR-NC and miR-134 mimic were transfected into A549 cells.The effect of miR-134 on proliferation of A549 cells was detected by methyl thiazolyl tetrazolium (MTT) and colony form experiment.Flow cytometry was used to determine the effect of miR-134 on A549 cells apoptosis.The effect of miR-134 on the expression of P53 protein was detected by Western blotting.Results The relative expressions of miR-134 in NSCLC tumor tissues and adjacent tissues were 0.429 ± 0.126 and 0.971 ±0.183 respectively,and the difference was statistically significant (t =7.742,P ＜0.001).The relative expressions of miR-134 in BEAS-2B cells and A549 cells were 1.013 ± 0.095 and 0.371 ± 0.068 respectively,and the difference was statistically significant (t =17.377,P ＜ 0.001).The absorbance (A) values of A549 cells transfected with miR-mimic were 0.451 ±0.051 and 0.518 ±0.074 on the third and forth day respectively,and those of A549 cells transfected with miR-NC were 0.683 ± 0.041 and 0.815 ± 0.065 respectively.The proliferation ability of miR-mimic group was significantly lower than that of miR-NC group (t =12.965,P ＜ 0.001;t =9.535,P ＜ 0.001).The colony forming rates of A549 cells transfected with miR-NC and miR-134 mimic were 91.2％ ± 8.3％ and 38.6％ ±4.5％ respectively,and the colony forming rate of A549 cells in miR-134 mimic group was significantly decreased (t =17.617,P ＜0.001).The apoptosis rates of miR-134 mimic group and miR-NC group were 93.5％ ± 3.7％ and 85.4％ ± 2.0％ respectively,and the difference was significant difference (t =6.119,P ＜ 0.001).The relative expressions of P53 protein in miR-134 mimic group and miR-NC group were 1.816 ±0.173 and 0.992 ± 0.096 respectively,and the difference was statistically significant (t =19.308,P ＜ 0.001).Conclusion miR-134 can be an effective target for the treatment of NSCLC by increasing the protein expression of P53,inhibiting the viability and proliferation of tumor cells,and promoting the apoptosis of tumor cells.