Impact of Intraoperative Blood Pressure Control and Temporary Parent Artery Blocking on Prognosis in Cerebral Aneurysms Surgery / 中国医学科学杂志(英文版)

Objective In cerebral aneurysm clipping and embolization, blood pressure control and temporary parent artery blocking are common methods to prevent aneurysm rupture. Their influence on the prognosis is uncertain. In this study, we try to find out the association between methods above and prognostic indicators.Methods We held a retrospective analysis on patients' medical records of cerebral aneurysms surgical clipping and endovascular coiling , and recorded gender, age, diagnosis, Hunt-Hess grade, Glasgow coma scale score, treatment methods, a history of hypertension, preoperative systolic blood pressure, with or without controlled hypotension, systolic blood pressure difference before and after controlled hypotension, with or without temporary artery blocking, with or without hypertension after treated aneurysm, prognostic indicators including mortality after 1 month, intensive care unit (ICU) stay time of survivors, discharged Glasgow outcome scale (GOS) score. Prognostic indicators were regarded as dependent variable, all the factors were regarded as independent variable, and the strength analysis of influence factors on prognostic indicators was made by binary logistic regression.Results Total cases were 165, including 68 males and 97 females, with an average age of 56 (12-85) years. The mortality after 1 month was 10.9% (18 cases). The ICU stay time of survivors was 7.35 (0-67) days. GOS score at discharge was 1-3 in 40 (24.2%) patients and 4-5 in 125 (75.8%) patients. Systolic blood pressure difference before and after controlled hypotension was an independent factor influencing mortality (t=2.273, P=0.024), and the greater the difference was, the higher the mortality would be. Timely hypertension after aneurysm treated was an independent factor affecting ICU stay time of survivors and patients with hypertension had shorter ICU stay time (χ=10.017, P=0.001). Blood pressure control (χ=0.088, P=0.767) and temporary blocking (χ=1.307, P=0.253) did not show significant influence on GOS score at discharge.Conclusions Timely controlled hypertension after aneurysm clipping and embolization can significantly shorten the stay time in ICU. The degree of controlled hypotension associates with postoperative mortality, the greater systolic blood pressure difference before and after antihypertensive treatment is, the higher the mortality will be.

Effect of the 18kDa translocator protein on U251 cells of human glioma / 中华神经医学杂志

Objective To study the effect of the 18kDa translocator protein (TSPO) on U251cells of human glioma. Methods U251 cell line was cultured in vitro conventionally.The specific ligand ofTSPO,pk11195,was used in 5 experimental groups respectively with concentrations of 100,50,25,12.5 and 6.25 μmol/L,in comparison with a control group.MTr colorimetry and trypan blue staining were used to detect cell proliferation.Hoechst33342 staining and flow cytometry were applied to detect cell apoptosis. Western blotting and immumofluorescence method were used to detect the expression level of TSPO. DCFH-DA probe and GSH kit were used to respectively detect the level of reactive oxygen species (ROS) and GSH level in cells.Jc-1 staining was applied to detect the mitochondrial membrane potential.Luciferase enzyme was used to detect the quantity of ATP in cells. Results MTT showed the survival of U251 cells was significantly higher in the groups of 50 and 25 μmol/L pk11195than in the control group (P＜0.05). Trypan blue staining showed the cell death rate was significantlylower in the group of 50 μmol/L pk11195 than in the control group (P＜0.05).The apoptosis rate,TSPO expression,ROS and GSH levels decreased significantly in the groups of 6.25 and 50 μmol/L pk11195,compared with the control group; the apoptosis rate was significantly lower in the group of 50 μmol/Lpk11195 than in the group of 6.25 μmol/L pk11195 (P＜0.05).The cell membrane potential and ATP quantity were significantly higher in the groups of 6.25 and 50 μmol/L pk11195 than in the control group,and those in the group of 50 μmol/L pk11195 were significantly higher than in the group of 6.25 μmol/Lpk11195 (P＜0.05). Conclusion TSPO may promote apoptosis of U251 cells in human glioma and inhibit proliferation of glioma cells,functioning similarly as a cancer suppressor gene.

The construction of lentivirus-mediated RNAi vector containing hTERT / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To construct a recombinant lentivirus RNA interference (RNAi) vector carrying hTERT gene, and to obtain the titer of the lentiviral stock for investigating the expression in the eukaryotic cells and the effect on the hTERT gene silencing in the eukaryotic cells.</p><p><b>METHODS</b>Two complimentary oligos of small interference RNA (siRNA) with hairpin structures targeting the hTERT gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced. The recombinant vectors were then transfected with viral packaging mix into T293 cells, viral supernatant was harvested to determine the titer. U87 cells infected by virus were harvested and the expression of hTERT, telomerase activity and apoptosis were detected by reverse transcription-PCR(RT-PCR), TRAP assay and flow cytometry separately.</p><p><b>RESULTS</b>Sequencing data showed that the constructed plasmids contained the correct sequences of hTERT siRNA transcript templates. A vector producing cell line T293 was established, and the titer for transfection was obtained. RT-PCR and TRAP flow cytometry analyses demonstrated that hTERT shRNA expression construct could suppress the expression of hTERT and telomerase activity and induce apoptosis.</p><p><b>CONCLUSION</b>A lentivirus RNAi vector targeting hTERT gene was successfully constructed, which decreased the expression of hTERT and telomerase activity effectively and induced apoptosis. It has set up a research platform for the gene therapy of tumors which take hTERT as the target.</p>

Study on combined gene therapy for malignant gliomas transfected with antisense hTERT/PTEN in vitro and in vivo / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.</p><p><b>METHODS</b>To construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.</p><p><b>RESULTS</b>After transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.</p><p><b>CONCLUSION</b>Growth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.</p>