Transplantation of bcl-2 gene-modified bone marrow mesenchymal stem cells improves cardiac function and angiogenesis in rabbit ischemic car-diac insufficiency model

AIM: To investigate the effects of transplantation of bone marrow mesenchymal stem cells (BMSCs) modified by bcl-2 gene on myocardial cell apoptosis, angiogenesis and cardiac function in the rabbit after acute myocardial infarction (MI).METHODS: The rabbit BMSCs were isolated, cultured and purified in vitro. The BMSCs were transfected with adenovirus or adenovirus-Bcl-2. The rabbit model of MI was established by ligation of left anterior descending branch. The rabbits were injected with Ad-Bcl-2-BMSCs (MI+Bcl-2-BMSCs group), Ad-BMSCs (MI+BMSCs group) and DMEM (MI group) in infarction marginal zone 2 weeks after ligation. The cardiac function was evaluated by echocardiography.The apoptosis of myocardial cells was measured by TUNEL. The mRNA expression of VEGF was detected by real-time PCR. The expression of CD31 was examined by immunohistochemical staining, and new blood capillaries were counted at 4 weeks after BMSCs transplantation. The correlation of the above values with cardiac function was analyzed.RESULTS: The cardiac function was better, the apoptotic rate was lower, the mRNA expression of VEGF and the capillary density were higher in both MI+Bcl-2-BMSCs group and the MI+BMSCs group than those in MI group, and those in MI+Bcl-2-BMSCs group increased more obviously.The left ventricular ejection fraction (LVEF) had a negative correlation with the myocardial cell apoptosis rate. A positive correlation with the mRNA expression level of VEGF and the capillary density was also observed.CONCLUSION: The transplantation of BMSCs modified by bcl-2 gene significantly reduces the myocardial cell apoptosis, promotes angiogenesis, improves heart function of the rabbits with MI.     

Protective effect of BNDF on vascular endothelial cells with H2O2-induced oxidative injury

AIM: To study the protective effect of brain-derived neurotrophic factor (BDNF) on vascular endothelial cells with H₂O₂-induced oxidative injury. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro, and the oxidation injury model of HUVECs was established by treatment with H₂O₂. The oxidatively injured HUVECs were cultured with different concentrations (1, 10 and 100 μg/L) of BDNF. At the same time, the control group (no injury), PBS treatment after H₂O₂ injury group and TrkB inhibitor group (with 100 μg/L BDNF and 1: 1 000 TrkB inhibitor) were also set up. The viability of the HUVECs was detected by MTT assay. The levels of LDH, MDA, SOD and GSH were measured. The releases of NO, ET-1 and ICAM-1 were analyzed by ELISA. The changes of ROS production and cell apoptosis were evaluated by flow cytometry. The protein levels of TrkB, p-TrkB, cleaved caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with uninjured control group, in H₂O₂ oxidative injury plus PBS treatment group, the viability of the cells was decreased significantly, the LDH and MDA levels were increased significantly and the activities of SOD and GSH were decreased significantly. The NO secretion was decreased, and the ET-1 and ICAM-1 concentrations were increased significantly. The ROS content and apoptotic rate were increased significantly. The protein levels of cleaved caspase-3 and Bax were increased but Bcl-2 protein expression was decreased significantly. Compared with PBS treatment group, in H₂O₂-injured HUVECs treated with different concentrations of BDNF, the cell viability was gradually increased, the LDH and MDA levels were decreased and the activities of SOD and GSH were increased gradually. The secretion of NO was increased but ET-1 and ICAM-1 were decreased gradually. The ROS content and apoptotic rate were decreased significantly. The TrkB and p-TrkB levels were significantly increased significantly, the protein expression of cleaved-caspase 3 and Bax was decreased gradually and the Bcl-2 protein expression increased gradually. The role of BDNF was inhibited by TrkB inhibitor. CONCLUSION: BDNF protects HUVECs from oxidative injury by binding with TrkB to activate the BDNF-TrkB signaling pathways.     

Sodium dichloroacetate improves radiosensitivity of U251 cells

AIM: To study the change of radiosensitivity of U251 cells after treated with sodium dichloroacetate (DCA) and further to explore the possible mechanism.METHODS: The U251 cells were divided into 4 groups: control group, DCA group, X-ray irradiation without DCA pretreatment (IR) group and X-ray irradiation with DCA pretreatment (DIR) group. MTT assay was applied to determine the cell viability. The intracellular reactive oxygen species (ROS) were detected by DHE fluorescence. The expression level of Bcl-2 was assessed by Western blot. The percentage of apoptosis of cells was determined by flow cytometry. RESULTS: No difference between control group and DCA group in cell viability (P>0.05) was observed. However, the cell viability of both IR group and DIR group was markedly reduced compared with control group (P<0.05). Furthermore, the viability of DIR group was significantly decreased compared with IR group (P<0.05). The percentage of ROS positive cells was obviously increased in DIR group compared with IR group (P<0.05). The expression level of Bcl-2 was sharply decreased in DIR group (P<0.05) and the percentage of apoptosis of cells was significantly elevated (P<0.05) in DIR group compared with IR group.CONCLUSION: The better antitumor effect was obtained by improving the radiosensitivity through pretreating the cells with DCA, and the possible mechanism was down-regulation of the Bcl-2 expression by developing the intracellular ROS.     

Hsa-miR-218 inhibits growth of cervical cancer HeLa cells by targeting to LASP1

AIM: To study the effect of hsa-miR-218 on cervical cancer HeLa cell growth and the underlying molecular mechanism.METHODS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed. pmiR-218 was transfected into HeLa cells. The number of viable HeLa cells was counted by the method of Trypan blue exclusion. The inhibitory rate of cell activity was detected by WST-8 assay. The expression of LIM and SH3 protein 1(LASP1) at mRNA and protein levels was determined by real-time PCR and Western blot. The interaction between miR-218 and LASP1 was examined using a luciferase reporter assay.RESULTS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed successfully and confirmed by DNA sequencing. Over-expression of miR-218 inhibited the activity of HeLa cells with the inhibitory rates of 15%, 26% and 65% at 24 h, 48 h and 72 h, respectively. The difference between transfection group and blank control/negative control group was statistically significant. The luciferase activity was reduced when co-transfection with miR-218 mimics and LASP1-3,UTR plasmid. The relative expression of miR-218 was increased after transfection with pmiR-218. Over-expression of miR-218 down-regulated the LASP1 expression at mRNA and protein levels by 25% and 75% respectively. Compared with blank control group and negative control group, the difference was statistically significant(P<0.05).CONCLUSION: pmiR-218 effectively inhibits the growth of HeLa cells in a time-dependent manner. miR-218 targets to the 3,UTR of LASP1, thus down-regulating the expression of LASP1 in HeLa cells.     

miR-21 inhibits apoptosis of Schwann cells following peripheral nerve injury

AIM: To explore the relationship and molecular mechanism between microRNA-21(miR-21) and Schwann cells (SC) following peripheral nerve injury. METHODS: The mRNA expression of miR-21 and phosphatase and tensin homologue deleted on chromosome ten (PTEN) in animal model were detected by real-time PCR. The over-expression of miR-21 and inhibition of miR-21 expression in the Schwann cells according to transfection of lentiviral vectors were performed, the nonspecific miRNA was used as a negative control (NC). The cell apoptosis was measured by flow cytometry. The mRNA expression of miR-21 and PTEN in the cells was detected by real-time PCR. The protein expression of PTEN and cleaved caspase-3 was determined by Western blot. RESULTS: The level of miR-21 was significantly higher and the mRNA level of PTEN was significantly lower in the model of nerve injury than those in control group. miR-21 over-expression decreased the number of apoptotic Schwann cells compared with NC-SC. The mRNA expression of PTEN was down-regulated by over-expression of miR-21. The protein expression of PTEN and cleaved caspase-3 was down-regulated by over-expression of miR-21(P<0.05). CONCLUSION: miR-21 may play an important role in the peripheral nerve injury through inhibiting apoptosis of Schwann cells by down-regulating the expression of PTEN.     

洋葱苯丙氨酸解氨酶基因AcPAL2的克隆与表达分析

利用简并PCR技术和RACE技术克隆得到了一条洋葱的苯丙氨酸解氨酶(PAL)基因全长cDNA序列。该cDNA序列全长2349 bp,编码长685个氨基酸残基的多肽序列,命名为AcPAL2。Blast分析表明该序列与虎眼万年青Galtonia saundersiae、野蕉Musa balbisiana的相似性均较高。Real-time PCR表达及花青素含量分析表明,红皮洋葱该基因表达量最大,而黄皮和白皮洋葱表达量极低;在红皮洋葱中该基因在膨大初期大量表达,并迅速降低至一定程度后趋于相对平稳表达,且与花青素的积累过程相一致。     

二化螟盘绒茧蜂寄生对寄主二化螟幼虫免疫反应的影响

为了探明二化螟盘绒茧蜂Cotesia chilonis (Matsumura)寄生对寄主二化螟Chilo suppressalis (Walker)幼虫血细胞和体液免疫反应的影响,观察并测定了二化螟盘绒茧蜂寄生引起二化螟幼虫血细胞数量、延展、存活、吞噬和包囊作用以及血淋巴酚氧化酶活性等的变化。结果显示,二化螟幼虫血细胞总数在寄生后1天即显著高于对照。其中,颗粒血细胞和浆血细胞在寄生后的数量变化均与血细胞总数变化相似,颗粒血细胞在寄生后3天、浆血细胞在寄生后0.5天起即分别与其对照差异显著,但两种血细胞在血细胞总数中各自所占的比例与对照多无显著差异。二化螟幼虫血细胞延展能力在寄生后0.5天受到显著抑制,此后与对照无显著差异;同时寄生可增加寄主血细胞的死亡率。寄主幼虫血细胞的吞噬作用在寄生后2天起显著降低;包囊作用则在寄生后1天起显著降低。二化螟幼虫被二化螟盘绒茧蜂寄生后,血淋巴酚氧化酶活性显著升高,但随后呈逐渐下降趋势。研究结果说明,二化螟盘绒茧蜂寄生使寄主二化螟幼虫的免疫反应呈现一定的规律变化。     

半闭弯尾姬蜂寄生对寄主小菜蛾幼虫体液免疫的影响

研究半闭弯尾姬蜂Diadegma semiclausum对寄主小菜蛾Plutella xylostella幼虫体液免疫系统的影响。结果表明,半闭弯尾姬蜂寄生能够显著抑制小菜蛾血淋巴内酚氧化酶前体激活,导致体外处理中血淋巴黑化的时间被延长;同时,抑菌物质的产生也受抑制并最终造成离体培养中血细胞的感菌率提高。     

Selection of Adequate Species for Degraded Areas by Oil‐Exploitation Industry as a Key Factor for Recovery Forest in the Ecuadorian Amazon

One of the requirements for the forest restoration of soils disturbed by the oil‐exploitation industry is that saplings be able to endure soil‐adverse conditions. In this study, saplings of 20 species susceptible to be used in reforestation programs were evaluated for their ability to grow on substrates derived from soils disturbed by petroleum extractions in the Ecuadorian Amazon. Seeds of each species were planted in germination trays. Once seedlings reached 5 cm in height they were transplanted to plastic bags with three treatment substrates: two derived from petroleum‐exploitation activity (soils from mud and drill cutting cells and from areas surrounding oil wells) and a control soil. Plant survival rate, stem height, and diameter were measured on a weekly basis until 14 weeks after transplantation, when we harvested the plants and also measured plant biomass and calculated the Dickson quality index for each species. Oil‐exploitation by‐product substrates impaired the performance of many saplings, with the substrate from mud and drill cutting cells being the one that most affected plant performance. Only saplings of five native species in the Amazon basin—Apeiba membranaceae, Cedrelinga cateniformis, Inga densiflora, Myroxylon balsamum, and Pouroma cecropiifolia—exhibited high or similar Dickson quality index values in all soil treatments and performed better than the rest. The use of these five species in remediation of soils disturbed by petroleum extraction in the Amazon basin could prove important because of their high potential to adapt to these disturbed sites. Copyright © 2016 John Wiley & Sons, Ltd.     

雷蘑胞外多糖抗肿瘤活性研究

从雷蘑AS 5.105深层发酵的滤液中分离得到胞外粗多糖CGP,以KM系S180荷瘤小白鼠为实验模型,用免疫器官重量法,进行高、中、低剂量CGP的腹腔注射实验。结果表明,CGP具有较高的抑瘤活性,并与剂量呈显著相关性;高剂量组与对照组相比较,小白鼠胸腺指数和脾脏指数均有显著增加,P<0.05;与低剂量组相比较,脾脏指数有显著增加,P<0.05。表明CGP能明显提高小白鼠的免疫功能。