Gene expression profile of oligodendrocytes in vitro in early stage after ionizing irradiation / 中华放射医学与防护杂志

ObjectiveTo characterize the gene expression in acute phase of irradiated oligodendrocytes (OL) in vitro.Methods The total RNA was extracted from irradiated OLs with 10 Gy by 6 MV X-rays at 1 and 4 h.The Affymetrix RAT 230 2.0 microarray were used to evaluate and screen the gene expression profile.The quantitative real-time RT-PCR was performed to validate the microarray results of selected myelin basic protein (MBP) and neural cell adhesion molecule 1 ( NCAM-1 ) genes.Results Compared with un-irradiated OLs,there were 1079 different expressed genes in irradiated cells.Those genes were classified in 79 categories based on the functional classification.Some familiar genes associated with OL cellular physiological process,apoptosis,cell cycle control,metabolism,cell communication and receptor binding were included.Compared with the microarray results,the coincidence rate of real-time RT-PCR was 91.7%.The down-regulation of MBP and up-regulation of NCAM 1 gene expression were confirmed.Conclusions Radiation-induced changes in gene expression in OLs took place in acute phase and influenced by time-course.The changes of MBP and NCAM1 gene expression may play a key role in the pathogenesis of radiation-induced demyelination.

Effect of signal transducers and activators of transcription 3 gene silencing by shRNA mediated by lentiviral vector for the treatment of colorectal cancer / 中华消化外科杂志

Objective To determine the effect of signal transducers and activators of transcription 3 (STAT3) gene silencing by shRNA mediated by lentiviral vector for the treatment of colorectal cancer. Methods The recombinant lentiviral vector pRNAT-shSTAT3, empty lentiviral vector pRNAT-GFP, and lentiviral packaging plasmids in supernatant were collected to transfect HT-29 cells for harvesting the HT-29-shSTAT3 cells and HT29-GFP cells. Fifteen male rats were divided into three groups (n = 5 ), and then they were inoculated with HT-29cells, HT-29-GFP cells and HT-29-shSTAT3 cells, respectively. Cell growth was assessed by MTT assay and the changes in cell cycle were detected by flow cytometry. The changes in microvessel density (MVD) of tumors were detected by immunohistochemistry. All data were analysed by one-way analysis of variance. Results The growth of HT-29-shSTAT3 cells was significantly suppressed compared with HT-29 and HT-29-GFP cells (F = 632.50,P ＜ 0. 05 ). The proportions of cells at the G0/G1 phase were 68.7% ± 2.9% in HT-29-shSTAT3 cells, 38.5% ±1.6% in HT-29-GFP cells and 38.7% ± 2.3% in HT-29 cells, with a significant difference among the three groups (F = 166.53, P ＜ 0.05 ). The MVDs of HT-29 cells, HT-29-GFP cells and HT-29-shSTAT3 cells were 29 ±5, 28 ±4 and 10 ±3, respectively, with a significant difference among the three groups (F=31.60, P ＜0.05). Conclusion STAT3 gene silencing by shRNA mediated by lentiviral vector can significantly inhibit the growth of colorectal cancer cells.

Diagnosis and treatment of cholelithiasis complicated with abdominal tumors:a report of 36 caseas / 中国普通外科杂志

Objective To study the causes of mis diagnosis and the experience of diagnosis and management for cholelithiasis complicated with abdominal tumors.Methods Clinical data of 36 cases of cholelithiasis complicated with abdominal tumors were analyzed retrospectively.Among them,24 were male,12 female.The age ranged from 48 to 82 year old.Each case was diagnosis as cholelithiasis and admitted to our hospital.Results In 36 patients,18 was diagnosed as complecated with abdominal tumors preoperatively;12 was diagnosed intraoperatively,while in 6 cases the tumor was misdiagmosed for 5 days to 3 months,finally the tumor was comfirmed by reoperation and pathology.Among the 36 cases,the tumors cluding 5 of carcinoma of gallbladder(13.9%),4 of cholangiocarcinoma(11.1%),3 of hepatoma(8.3%),6 of pancreatic carcinoma(16.7%),6 of gastric carcinoma(16.7%),7 of colon carcinoma(19.4%),2 of rectal carcinoma(5.5%).Of them,2 cases refused operation,2 cases underwent intervention operation,the others reseived operation.During primary operation,radical resection of the tumor and cholecystectomy and/or common bile duct(CBD) exploration was performed in 18 patients,palliation resection and cholecystectomy and/or CBD exploration performed in 4 patients,exploratory laparotomy performed in 4 patients,and only LC and/or CBD exploration performed in 6 patients.Of the latters,reoperation was performed 5 days to 3 months after primary operation.4 patients had tumor radical resection,1 had palliation resection,and 1 had interventional therapy.Conclusions Cholelithiasis may complicated with abdominal tumor,especially with digestive tract tumor.For cholelithiasis patient the history-taking need to be done carefully before operation.For the senile patient and the patient without typical symptoms or physical signs,the systems checking shoud be done carefully preoperatively,and abdominal cavity should be checked carefully intraoperatvely to find the tumor,which may exist in abdominal cavity.the patient with syndrome of post-cholecystectomy shoud be carefully followed up to avoid the mis diagnosis and mistreatment of the tumor.

Changes of caspase-3 activities in rat hippocampal neurons during hypoxia/reoxygenation / 第二军医大学学报

Objective: To study caspase-3 activities and the neuroprotective effect of caspase-3 inhibitor Ac-DEVD-CMK in primary cultures of rat hippocampal neurons during hypoxia/reoxygenation (H/R). Methods: After pretreated with Ac-DEVD-CMK or the vector DMSO fo 1 h, primary cultures of rat hippocampal neurons were induced to hypoxia for 3 h, followed by 12 to 48 h of reoxygenation. The neuronal viability was estimated by MTT assay, and caspase-3 cellular activities were measured by a colorimetric method using Ac-DEVD-pNA as a substrate. Results: During H/R, the cultured rat hippocampal neuronal viability gradually decreased, and caspase-3 activities in the primary cultures of rat hippocampal neurons were significantly increased, and peaked at 24 h of reoxygenation. Caspase-3 activities were decreased in the neurons pretreated with Ac-DEVD-CMK in a dose dependent manner. Conclusion: Delayed neuronal death is detected in cultured hippocampal neurons during H/R, and apoptosis mediated by caspase-3 may play an important role in it, and caspase-3 inhibitor has a neuroprotective effect on them.

Caspase-3 expression on MCA ischemia-reperfusion in rat models / 临床神经病学杂志

Objective To study the role of caspase 3 in ischemic brain damage of rats, and further understand the molecular mechanisms of ischemic cerebral vascular disease.Methods Rat models of the left middle cerebral artery (MCA) occlusion/reperfusion were made using a modification of the intraluminal sature method of Longa established by Belayev, infarct zones were confirmed by 2,3,5 triphenyltetrazolium chloride (TTC) staining, and caspase 3 expression on brain sections at the mRNA and active protein level was detected with in situ hybridization and immunohistochemistry technique, respectively.Results After 2 hours of left MCA ischemia followed by 24 hours of reperfusion, obvious infarct in the MCA dominate regions was confirmed by TTC staining; low levels of caspase 3 mRNA, and fewer of its active protein expression was found in normal brains, sham brains and contralateral brains of MCAO rats; both caspase 3 mRNA and activated protein expression in ipsilateral region were increased after 24 hours of recirculation, and even higher levels were detected at 48 hours of reperfusion.Conclusion Apoptotic mechanism might involve in delayed neuronal death after cerebral ischemia, and caspase 3 might play an important role in ischemic neuronal injury.

Protective effects of ligustrazine on glutamate-induced neurotoxicity in cultured hippocampal neurons / 西安交通大学学报(医学版)

Objective To observe the protective effects of ligustrazine on glutamate-induced injury in cultured hippocampal neurons. Methods Primarily cultured hippocampal neurons from fetal rats were incubated with ligustrazine (1 mmol/L) for 12 hours,then glutamate (1 mmol/L) was added for 20 minutes to induce injury. Cell viability was detected by MTT assay,and the change in LDH activity was determined by biochemical method. Finally,the expression of AchE in the cultured neurons was assayed by immuocytochemistry. Results Ligustrazine improved the survival rate of hippocampal neurons injured by glutamate,inhibited the release of LDH from the kytoplasm injured by glutamate. The expression of AchE in the injured neurons was promoted by pretreatment of ligustrazine. Conclusion Ligustrazine can significantly exert protective effects on the hippocampal neuron injury induced by glutamate.