ObjectiveTo confirm the effect of miR-93 inhibitor in glioma cell growth and invasion.MethodsMalignant glioma cells were transfected with miR-93 inhibitor by lipofectamin to downregulate their overexpression of miR-93.Real time-PCR was taken to measure miR-93 expression after transfection.The cell cycle kinetics and cell growth rate were detected by flowcytometry and MTT assay,the cell proliferative ability was evaluated by soft agar assay,and the invasive ability was detected by transwell assay.ResultsThe highlevel expression of miR-93 was downregulated effectively in glioma cells after transfecting the miR-93 inhibitor.Meanwhile,the cell cycle progress was delayed,S phase cells were reduced,the speed of growth was slowed,cloning formation ability was receded,the number of cells through the matrigel was reduced,and invasive ability was significantly repressed.ConclusionDownregulation of miR-93 expression could inhibit the proliferative ability and invasive ability of glioma cells.

Objective To study the effects of postoperative immune-enhancing enteral nutrition (IEEN)on the liver function and postoperative infectious complications after hepatectomy in patients with hepatocellular carcinoma.Methods Between January 2010 and December 2011,96 patients undergoing hepatectomy for hepatocellular carcinoma were randomized in a double-blind manner to receive postoperative enteral nutrition with IEEN (IEEN group,n =48) or standard enteral nutrition (control group,n =48) for 5 days,respectively.Calorie and nitrogen of the same quantity were provided for both groups from the first day after surgery.The liver function tests and routine blood tests were performed.The postoperative hospital stay and postoperative infectious complications were also compared between these two groups.Results The serum albumin,prealbumin,alanine aminotransferase,and total bilirubin levels had no significant differences between the two groups (all P ＞ 0.05).On the 6th day after operation,the level of total lymphocyte count was significantly higher in the IEEN group than that in the control group [(2106 ± 1057) vs.(1648 ± 1079) / μL,P =0.038).The postoperative hospital stay was (10.3 ±5.6) days in the IEEN group and (13.4 ±6.7) days in the control group (P =0.016).Postoperative infectious complications were observed in 8 (16.7％) patients in the IEEN group and 17 (35.4％) patients in the control group (P =0.036).Conclusion Early postoperative IEEN can improve the clinical outcomes of liver cancer patients after hepatectomy.

OBJECTIVETo investigate molding technology of total alkaloids from Sophora alopecuroides freeze-dried powders and observe its inhibition effects on liver transplantation tumor in mice.

METHODWith color, clarity, water-soluble and formability as indexes, single factor tests were adopted to screen type and amount of filler, the concentration of total alkaloids in drug liquid, pH in order to determine optimum prescription of total alkaloids from S. alopecuroides freeze-dried powders, the lowest melting point was determined and freeze drying curve was drafted. Mice hepatoma H22 ascites tumor strain was sterile inoculated in right axillary subcutaneous of mice, and antitumor effect of total alkaloids from S. alopecuroides freeze-dried powders on liver transplantation tumor H22 in mice.

RESULTWhen selected 80 g x L(-1) as mannitol as filler, the concentration of total alkaloids in drug liquid was 25 g x L(-1) and pH 6.5-7.5, freeze-dried effect was optimum with fast reconstitute speed. Average inhibition rate of the big (120 mg x kg(-1)) and medium (60 mg x kg(-1)) dose group of total alkaloids from S. alopecuroides freeze-dried powders on liver transplantation tumor H22 in mice were 56.08% and 35.49%, respectively.

CONCLUSIONPreparation technology was reasonable, reproducible and stable, total alkaloids from S. alopecuroides freeze-dried powders had significant antitumor effect and showed a dose-effect relationship.

Alkaloids ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; Cell Line, Tumor ; Color ; Freeze Drying ; Humans ; Liver Neoplasms ; drug therapy ; Mice ; Powders ; chemistry ; Sophora ; chemistry ; Tumor Burden ; drug effects

Objective To evaluate the feasibility and therapeutic effect of kyphoplasty in treating severe osteoporotic vertebral compressive fractures. Methods Thirty-five patients (48 vertebral bodies) with severe osteoporotic compressive fractures were included. There were 33 females and 2 males with the mean age of 74.2 years. The average compressive rate of the affected vertebral bodies was 77.0%. The thoracolumbar vertebrae were treated with kyphoplasties. Percutaneous puncture direction was adjusted according to compressive rate and shape of the vertebral bodies. The inflatable bone tamp was inserted into the fractured vertebral body. The balloon was inflated with low pressure and dilate-relieve-dilate method was applied. The balloon was deflated and withdrawn, leaving a cavity within the vertebral body, which then fulfilled with visualized bone cement. Preoperative and postoperative symptom level, complications and radiographic findings were recorded. Results All 35 patients tolerated procedure well. The mean heights of the anterior, mid and posterior vertebral body had improved from (0.8±0.1) cm, (0.8±0.2) cm, (2.1 ±0.8) cm preoperatively to (1.2±0.3) cm, (1.3±0.2) cm, (2.3±1.0) cm respectively after operation (P ＜0.05). There was significance difference between preoperative and postoperative heights of the anterior and mid vertebral body. The mean kyphosis was improved from 28.2°±5.2° before operation to 19.1°±4.9° after operation. Conclusion Kyphoplasty is feasible and effective for severe osteoporotic vertebral compressive fractures.

300mmH2O), non-purulent CSF with pleocytosis up to 2700?106/L, increased polykaryocytes in the early stage, normal or obviously high total protein level, normal glucose and chloride level, and negative laboratory examinations (such as culture-and smear-examination of Mycobacterium tuberculosis, adenosine deaminase, anti-TB antibody, immuno-dot-blot assay MycoDot and PPD assay). In this series of 167 patients, 7 of them died (3.7%). Conclusions The findings in CSF examination in most patients with tuberculous meningitis may show atypical findings, therefore it should be emphasized to repeat the analysis of CSF in order to obtain an accurate diagnosis of tuberculous meningitis.

Objective To compare clinical features,pulmonary functions,chest imaging and prognosis between combined pulmonary fibrosis and emphysema syndrome (CPFE) and idiopathic pulmonary fibrosis(IPF) without emphysematous changes in elderly patients.Methods 88 elderly IPF patients in Beijing hospital from January 2000 to October 2012 were divided into CPFE (n=30) and IPF (n =58) groups according to the CT imaging.Clinical features,blood gas analysis,pulmonary function,chest CT and survival time were compared between the two groups.Results 30 CPFE patients with the mean age of(75.5 ±7.6) years and 58 IPF patients without emphysema with the mean age of(73.7±6.8) years were enrolled.The proportions of male patients,smoking history and mortality were higher in CPFE patients than in IPF group(86.7％ vs.63.8％,28 vs.36,76.7％ vs.43.1％,x2 =5.09,9.74,8.98,P＜0.05 or 0.01).CPFE patients had a higher force vital capacity(FVC) and total lung capacity(TLC) as compared with IPF group [(2.6±0.9) L vs.(2.1± 0.5) L,(5.4±1.9) L vs.(4.4±1.1) L,t=2.69,2.35,P＜0.01 or 0.05].There were no significant differences in forced expiratory volume in one second(FEV1) and the diffusion capacity for carbon monoxide(DLCO) between the two groups.The main type of emphysema by HRCT scan were centrilobular emphysema in CPFE patients.There were lower median survival time in CPFE group than in IPF group [(3.0±0.2) years vs.(4.0±1.0) years,x2=4.50,P＜0.05].Conclusions The majority of elderly CPFE patients are males and smokers.The lung volume is increased in elderly CPFE patients as compared with IPF patients.Centrilobular emphysema is the primary type in CPFE patients.The prognosis is worse in elderly CPFE patients than in IPF patients.

Objective To study the inhibitory effect of knocking down miR-30a-5p on the U87 human glioma xenograft growth and its possible mechanism.Methods Nude mice bearing subcutaneous U87 human glioblastoma were established and separated into three groups (eight for each group) by randomized digital table method,including control group,scr-ODN treated group and AS-miR-30a-5p treated group.After relevant subcutaneous injection treatment,tumor size was measured every other day until the observation period ended.Researchers executed the animals after the treatment,stripped tumor tissues and extracted RNA and protein.Real-time PCR was conducted to detect the expression of miR-30a-5p.The histopathological characteristics and proliferation and apoptosis biological characters (including SEPT7,PCNA,cyclin D1,MMP-2,apoptosis related factor P53,bcl-2 and caspase3) were evaluated by HE and immunohistochemical staining,Westem blot analysis respectively,and the cell apoptosis was detected by TUNEL method.Results In AS-miR-30a-5p treated group,the tumor growth was delayed and the final tumor volume was smaller than that in the control and scr-ODN treated group (F =7.167,P ＜0.05),and the expression of miR-30a-5p was knocked down.The expression of PCNA,cyclin D1 were significantly downregulated while P53,SEPT7 and caspase3 up-regulated.Apoptotic index was increased significantly.Conclusion As-miR-30a-5p suppresses the growth of U87 human gliomas xenografts significantly.Malignant phenotype of tumors are reversed to a considerable degree.Therefore,miR-30a-5p can be a candidate for targeted therapy of human glioma.

Objective To construct and confirm the JC virus(JCV) T antigen expression plasmid using mouse keratin 19 (K19) promoter specific for the gastric epithelial cells.Methods The Ndel site was mutated by FCR with Bell insertion at both sides.The DNA fragment digested by Bcl Ⅰ was ligated with the plasmid containing K19 promoter via Bam Ⅰ site.The DNA sequence was confirmed by restriction enzyme digestion and direct DNA sequencing.Cytokeratin 19 protein was examined to screen gastric carcinoma cell for transfection of K19-JCV T antigen expression plasmid by immunohistochemistry.The Western blot was employed to detect the JCV T antigen expression in the gastric carcinoma transfectant.Results K19-JCV T antigen expressing plasmid was successfully constructed.The ACS strongly expressed cytokeratin 19 protein and was selected for the transfection of K19-JCV T antigen expressing plasmid.JCV T antigen was positively expressed in the AGS transfectant.Conclusion The synonymous mutation and compatible ligation are useful in the plasmid construction.The methy lation of restriction enzyme should be considered.It is meaning for the transgenic animal model of gastric carcinoma to successfully construct the JC virus T antigen expression plasmid in gastric mucosa.

objective To identify SEPT7as one of the target genes of miR-19a and miR-19b.MethodsmiR-19a inhibitor and miR-19b inhibitor mediated by lipofectamine2000,were transfected to SNB19 glioma cells for knocking down miR-19a/19b overexpression.Real time PCR was conducted to detect the expression of miR-19a/miR-19b in transfected cells.The expression of SEPT7was determined by Western blot analysis.RT-PCR was used to detect the mRNA expression of SEPT7; and Luciferase reporter assay was used to identify the direct regulation of miR-19a/19b on SEPT7.ResultsIn SNB19 glioma cells transfected with miR-19a/19b inhibitor,the expression of miR-19a/miR-19b was significantly reduced,whereas the protein expression of SEPT7 was upreguhtted; no significant change of SEPT7 mRNA level was found and luciferase activity became stronger as compared to control cells.ConclusionSEPT7 is the target negatively regulated by miR-19a and miR-19b.

Objective To evaluate the effect of propofol on interleukin-1β (IL-1β)-induced increase in monolayer permeability of human umbilical vein endothelial cells (HUVECs).Methods Primary HUVECs were cultured and purified by immuno-magnetic separation.The expression of VE-cadherin in endothelial cells was determined by immunofluorescence.The HUVEC monolayer permeability was detected by the Transwell system.The cells were seeded on the upper chamber (2 × 105 cells/well) and cultured for 3 days after confluence.The cells were treated in two ways.The cells were randomly divided into 6 groups (n =36 each) and 5 of the 6 groups treated with 1,2,5,10 and 20 ng/ml IL-1β for 24 h except for control group.The cells were also randomly divided into 5 groups (n =30 each) and 4 of the 5 groups were pretreated with 0,10,50 and 100 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h except for control group.The cells were radomly divided into 3 groups (n =18 each) and 2 of the 3 groups were pretreated with 50 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h or 30 min.The expression of occludin protien,p38 mitogen activiated protienkinase (p38 MAPK) and phosphorylated p38 MAPK (p-p38 MAPK) was determined by Western blot.Results Compared with control group,5,10 and 20 ng/ml IL-1β significantly increased HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.05 or 0.01).10,50 and 100 μmol/L propofol inhibited IL-1 β-induced increase in the permeability of HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.01).IL-1β could down-regulate HUVEC occludin protein expression,and activate p38MAPK signaling pathway,and propofol inhibited IL-1β-induced down-regulation of HUVEC occludin protein expression and activation of p38 MAPK signaling pathway (P ＜ 0.01).Conclusion Propofol can alleviate IL-1β-induced increase in the permeability of HUVEC monolayer via inhibiting activation of p38 MAPK signaling pathway.