1.A study of downregulation of miR-93 on the suppression of human glioma cell growth and invasion
Anling ZHANG ; Kun WANG ; Guangxiu WANG ; Zhifan JIA ; Peiyu PU
Journal of International Oncology 2012;39(1):68-72
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.
2.Effect of postoperative immune-enhancing enteral nutrition on outcomes after hepatectomy in patients with hepatocellular carcinoma
Yonghui SU ; Bingzong HOU ; Yingbin JIA ; Xiaoling LI ; Juyuan PU
Chinese Journal of Clinical Nutrition 2012;20(5):302-305
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.
3.Treatment of elderly severe osteoporotic vertebral compressive fractures by kyphoplasty
Hai TANG ; Hao CHEN ; Bingqiang WANG ; Jinjun LI ; Pu JIA
Chinese Journal of Orthopaedics 2010;30(10):978-983
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.
4.Preparation of total alkaloids from Sophora alopecuroides freeze-dried powders and investigation of its antitumor effects.
Jia-Fu YAN ; Yan TONG ; Chun PU ; Jin-Yu WANG
China Journal of Chinese Materia Medica 2014;39(7):1234-1237
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
5.Analysis of cerebrospinal fluid from 167 cases of tuberculous meningitis
Jia HU ; Jiatang ZHANG ; Senyang LANG ; Chuanqiang PU
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
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.
6.Inhibitory effect of antisense miR-30a-5p on glioma cell growth in vivo
Jikui SUN ; Zhifan JIA ; Peiyu PU ; Guangxiu WANG ; Anling ZHANG ; Weidong YANG
Journal of International Oncology 2013;(2):149-152
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.
7.The Construction and Expression Confirmation of JC Virus T Antigen Expression Plasmid in Gastric Mucosa
Pu XIA ; Xiaoyan XU ; Baoping JIA ; Wei WANG ; Yifu GUAN ; Yasuo TAKANO ; Huachuan ZHENG
Journal of China Medical University 2010;(1):18-21
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.
8.Regulation of miR-19a and miR-19b on tumor suppressor gene-SEPT7
Kun WANG ; Guangxiu WANG ; Peiyu PU ; Anling ZHANG ; Jianwei HAO ; Zhifan JIA
Journal of International Oncology 2011;38(3):233-236
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.
9.QUORUM SENSING IN PLANT-ASSOCIATED BACTERIA
Shui-Shan SONG ; Zhen-Hua JIA ; Zhen-Xian GAO ; Hong MA ; Pu-Fan DUAN ;
Microbiology 1992;0(02):-
Bacterial N-Acyl-L-homoserine lactones (AHLs)-mediated quorum sensing is involved in the regulation of diverse biological functions. As the bacterial pathogen population density increases, AHLs concentration secreted by pathogen reaches a threshold and then interacts with their intercellular receptor and triggers expression of virulence genes. It is a promising approach to biologically control bacterial disease in plants and animals by manipulating bacterial AHL-quorum sensing with AHLs-degrading enzyme and AHL analogue.
10.Host-virus Interaction at the miRNA Level
Yu-Shu ZHENG ; Pu ZHAO ; Bei-Bei JIA ; Xing-You LIU ;
Microbiology 2008;0(07):-
MicroRNAs (miRNAs) are recently discovered major regulators of gene expression, which play a pivotal role in a wide spectrum of biological processes including antiviral defence. There is growing evidence that some viruses either encode their own viral miRNAs or subvert cellular miRNAs. The host-and virus-encoded miRNAs and their targets together thus form a novel regulatory layer of interactions between the host and the virus. A better understanding of host-virus interaction mediated by miRNAs would not only enable us to unravel the molecular basis of viral pathogenesis, but also enable us to develop better therapeutic strategies.