1.Expression and significance of signaling pathway of miR-134/CREB/pCREB in patients with epilepsy and in epileptic rats
Qian WANG ; Yangmei CHEN ; Jing GUO ; Xiaolan YANG ; Yunlan XIE
Acta Laboratorium Animalis Scientia Sinica 2014;(6):28-33
Objective To investigate the expression of microRNA-134 ( miR-134 ) , CREB and pCREB in the temporal lobe tissue of patients and epileptic rats and to explore their roles in pathogenesis of epilepsy.Methods Tempo-ral lobe tissue samples of 14 patients with refractory epilepsy and 10 non-epileptic patients, and hippocampus and brain tis-sue samples of 42 rats were used in this study.Forty-two healthy adult male Sprague-Dawley rats were randomly divided in-to 6 epilepsy groups (24 h, 72 h, 7 d, 14 d, 30 d, and 60 d after kindling epilepsy) and a normal control group (n=6 for all groups) .The rat model of epilepsy was generated by intraperitoneal injection of 127 mg/kg lithium chloride and 16-20 h later, 35 mg/kg pilocarpine.In the temporal lobe tissue of patients and hippocampal tissue of rats, the expression level of miR-134 was detected by real-time polymerase chain reaction.The expression levels of CREB and pCREB were de-termined by Western blot, and CREB and pCREB localization was assessed by immunohistochemistry.Results Compared with the control rats, the expression of miR-134 was significantly decreased in the temporal lobe tissue of experimental rats at 72 h,7 d,14 d, 60 d after kindling (P<0.05),and no significant change at 24 h and 30 d after kindling (P>0.05). Expression of miR-134 in patients with refractory epilepsy was significantly lower than that of the controls ( P<0.05 ) , while up-regulation of CREB expression was at the same time points (P<0.05).Up-regulation of pCREB expression was at all the time points after kindling (P<0.05).CREB and p-CREB expressions were seen in the nuclei of neurons, and significantly higher in patients with refractory epilepsy and epileptic rats.Conclusions The expression of miR-134 is sig-nificantly decreased and that of CREB and pCREB was significantly increased in the temporal lobe tissue of patients with re-fractory epilepsy and the hippocampal tissue of epileptic rats.These findings indicate that the signaling pathway of miR-134/CREB/pCREB may play an important role in the pathogenesis of epilepsy.
2.Clinical observation of three dimensional conformal radiation therapy in combination with chemotherapy in stage Ⅲ patients with non-small-cell lung cancer
Xiaomin LI ; Yunlan LI ; Runhua AN ; Dan GUO ; Xia HOU
Cancer Research and Clinic 2008;20(5):322-323,326
Objective To observe the toxicity and effection of three dimensional conformal radiation therapy(3DCRT)based on conventional fractionation radiotherapy in combination with chemotherapy in non-small-cell lung cancer(NSCLC).Methods 62 NSCLC patients with stage Ⅲ were chosen as research objects.All the patients received chemotherapy two cycles with NP regimen firstly(NVB 40 mg d1,8,DDP 30 mg d2-5),then they accepted radiotherapy in twentith day after chemotherapy when hemocyte got normal.In the first half period of the radiotherapy,conventional radiotherapy were given with 2 Gy/f,after the dose got 40 Gy/20 f,then three dimensional conformal radiation therapy were given with 4~6 Gy/f every 1 to 2 days until the total dose got 24~30 Gy/4~6 f.The patients with supraclavicular lymphnode metastatic carcinoma received conventional fractionation radiotherapy with 60Co or 6 MV-X ray combined electronic ray until the dose(DT)up to 64~66 Gy.After radiotherapy,the patients accepted chemotherapy two cycles with NP regimen again.Results All the 62 patients completed treatment plan,and follow-up survey had lasted for 3 years.The short-term efficacy was evaluated in 3 months after radiotherapy. Among 62 patients, 9 cases gained a complete remission(CR),40 cases partial remission(PR),and the total remission was 79.0%.The acute toxicity showed acute radiation esophagitis,radiation pneumonia and acute gastrointestinal reaction.However,expectant treatment was effective.The 1,2,3 year survival rate was 7 1.0%,48.4%,30.6%,respectively.The 1,2,3 year local control rate was 80.6%,62.9%,40.3%,respectively.Conclusion 3DCRT based on conventional fractionation radiotherapy for NSCLC have good effection and mild toxicity.
3.Effects of silencing RPB5-mediating protein (RMP) gene on cell proliferation and migration of liver cancer SMMC-7721 cells
Xiaoning LIAN ; Huicui YANG ; Kai CAO ; Min LI ; Weihua SHENG ; Xiaoting WANG ; Yunlan GUO ; Wenxiang WEI
Tumor 2010;(1):15-20
Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.
4.Non-fermenter isolation and drug resistance analysis of clinical specimens from the year of 2010 to 2012
Yunqi GUO ; Yunlan GUO ; Yi WANG ; Lihong PEI ; Xuepeng GUO
Chinese Journal of Applied Clinical Pediatrics 2014;29(4):295-298
Objective To study the non-fermenter isolation rate and its drug resistance to more than 10 kinds of antibiotics at the First Affiliated Hospital to Xinxiang Medical University from the year of 2010 to 2012 in order to offer evidences for reasonable use of drugs by doctors and to prevent nonfermenters from becoming common pathogenic bacteria.Methods The infection and drug resistance of different Nonfermenters (Pseudomonas aeruginosa,Acinetobacter baumannii,Pseudomon as mahophilia,Los non Acinetobacter,Pseudomonas stutzeri,Achromobacter,Bacillus steady short,onions Bock cepacia,meningitis septicemia Elizabeth Kim bacteria)during present 3 years were analyzed by using WHONET 5.5 software.Results The infection rate from non-fermenters as pathogenic bacteria was gradually increased,with the rate from 18.2% to 27.2%,until 31.2% ; Acinetobacter Baumannii were isolated as higher infection rate and multiple drug resistance (7.1% to 18.5% until 28.7%) in pathogenic bacteria; the infection rate of other non-fermenter with infection rate of high and multiple drug resistance in pathogenic bacteria were gradually increased,such as infection rate or the precedence of Achromobacter xylosoxidans and Pseudomonas maltophilia,especially the precedence of Pseudomonas maltophilia ranged the third in non-fermenters,and Pseudomonas maltophiliai had a higher and multiple drug resistance to many antibiotics; infection rate of non-fermenters with low drug resistance gradually decreased,such as Pseudomonas fluorescens; the drug resistance rate of Acinetobacter Baumannii was higher than that of Pseudomonas aeruginosa and its drug resistance rate was over 50% to common antibiotics used clinically,as well as Pseudomonas maltophiliai and Empedobacter brevis,especially the resistance rate of Pseudomonas maltophiliai to many commonly used antibiotics was more than 90.0%.Conclusions Non-fermenter isolation rate is becoming higher clinically and its drug resistance becomes more severe,so the sanitation administration department should pay more attention to the use of antibiotics in order to prevent and control drug resistance and higher infection rate caused by non-fermenters as a serious consequence.