miR-451 inhibits inflammatory responses in glomerular mesangial cells by targeting Psmb8 in diabetic nephropathy mice

AIM: To investigate the effect of microRNA (miR)-451 by targeting proteasome subunit β type 8 (Psmb8) on the inflammatory responses in mouse glomerular mesangial cells (MCs) under high-and low-glucose conditions. METHODS: The expression levels of miR-451, IL-18 mRNA and TNF-α mRNA were detected by qPCR. The protein expression levels of IL-18, TNF-α and Psmb8 were determined by Western blot when miR-451 was over-expressed and down-expressed in the MCs. Moreover, the expression of IL-18 and TNF-α was detected when Psmb8 was silenced by si-Psmb8 in MCs. RESULTS: The expression of miR-451 was significantly decreased in the MCs treated with high glucose compared with low glucose group (P<0.01). However, the expression of Psmb8 was increased in high glucose group as compared with low glucose group (P<0.01). Moreover, the expression levels of Psmb8, IL-18 and TNF-α were significantly decreased when miR-451 was over-expressed in high glucose group (P<0.01). Additionally, the expression levels of IL-18 and TNF-α were significantly reduced when Psmb8 was silenced in the MCs under high glucose condition. CONCLUSION: miR-451 reduces the expression of inflammatory factors via targeting Psmb8 in the MCs under high glucose condition. Therefore, miR-451 may play a role in inflammation of diabetic nephropathy.     

Long non-coding RNA PVT1 regulates effect of miR-551 on migration and invasion abilities of ovarian cancer through Wnt signaling pathway

AIM: To investigate the expression of long non-coding RNA PVT1 in ovarian cancer and the role of PVT1 in migration and invasion abilities of ovarian cancer cells.METHODS: The expression of PVT1 in ovarian cancer tissue, normal ovarian tissue and different ovarian cancer cell lines was detected by qPCR. Transwell assay was used to detect the invasion ability of ovarian cancer cells after PVT1 silencing. The migration ability of the ovarian cancer cells after PVT1 silencing was detected by scratch test. The interaction between PVT1 and microRNA (miR)-551 was analyzed by dual-luciferase reporter assay. The effect of miR-551-inhibitor on the invasion and migration abilities of ovarian cancer cells after PVT1 silencing was detected by Transwell assay and scratch test. The expression of Wnt signaling pathway-related proteins was determined by Western blot after PVT1 silencing. The effects of PVT1 silencing on tumor weight and volume of ovarian cancer were examined by subcutaneous tumor transplantation in nude mice.RESULTS: The expression of PVT1 in ovarian cancer tissue was significantly higher than that in normal ovarian tissue (P<0.05). The expression level of PVT1 in ovarian cancer cell line ES-2 was the highest. PVT1 silencing inhibited the invasion and migration abilities of the ovarian cancer cells. After PVT1 silencing, miR-551-inhibitor promoted the invasion and migration abilities of the ovarian cancer cells. The expression of Wnt signaling pathway-related proteins was decreased after PVT1 silencing (P<0.05). Compared with negative control group, the tumor volume and weight in PVT1-siRNA group were significantly decreased (P<0.05).CONCLUSION: PVT1 plays an important role in the development of ovarian cancer. PVT1 regulates the invasion and migration abilities of ovarian cancer cells through Wnt signaling pathway.     

籼稻品种93-11遗传转化中抗生素适宜剂量的筛选

为优化农杆菌介导转化体系,研究了两种常用抑菌剂头孢霉素(cef)和羧苄青霉素(cb)的抑菌效果和对籼稻品种93-11成熟胚培养的影响,并进一步研究选择剂潮霉素(hyg)对愈伤诱导、分化和种子萌发生根的影响,确定其作为选择剂的有效浓度.对cb和cef共设置12种质量浓度处理,各处理都能完全抑制农杆菌的生长,同时对愈伤的诱导和生长没有不良影响,但是有些处理使愈伤的分化率下降.150、250 mg.L-1cb的单独作用,400、500 mg.L-1cef的单独作用,以及cb为100mg.L-1时,300、400、500 mg.L-1cef的3种互作处理都不影响愈伤的分化率;但cef高达600 mg.L-1时明显降低愈伤的分化率,达极显著水平,使分化率降到7%;cb为50 mg.L-1时,350、450、550 mg.L-1cef的3种互作处理都极显著地降低愈伤的分化率,表明高浓度的cef以及添加小量cb的处理与cef之间的互作对分化率的影响十分显著.两种抑菌剂单独使用(400-500 mg.L-1cef,150-250 mg.L-1cb)即有效抑制转化和培养过程中农杆菌的繁殖,同时不影响愈伤的生长和分化.对hyg的研究表明,愈伤诱导和分化对hyg最为敏感,5 mg.L-1的剂量足以使出愈率为0,使愈伤的分化率为0;hyg对愈伤的致死剂量为40 mg.L-1;hyg对种子萌发生根的致死剂量为25 mg.L-1.因此,在愈伤分化阶段不宜加入选择剂;在抗性愈伤组织筛选阶段加入hyg的适宜质量浓度要大于40 mg.L-1;利用愈伤诱导和种子萌发生根对hyg的敏感性,可用于转基因种子的筛选,筛选质量浓度分别要大于5和25 mg.L-1.     

玫瑰黄链霉菌Men-myco-93-63活性成分的初步研究

对拮抗链霉菌Men-myco-93-63固体培养基中活性成分的提取分离进行了初步研究,最后确定乙酸乙酯与丙酮3∶1（V/V）的混合提取剂为最佳提取剂,并明确了该活性成分属于脂溶性。进行了活性成分存在部位的测定试验,结果发现活性成分既存在于菌丝体中,又存在于培养基中。通过HPLC对活性成分进行检测,确定了其中能够进行纯化的组分。     

玫瑰黄链霉菌Men-myco-93-63发酵液对黄瓜白粉病抗性的影响

药效试验表明,玫瑰黄链霉菌Men-myco-93-63发酵液对黄瓜白粉病有良好的防效,其中发酵液保护作用的防效为88.63%,治疗作用的防效为82.26%,表明该发酵液对黄瓜白粉病的防治既有保护作用又有治疗作用。通过对喷施发酵液后黄瓜叶片内的抗病相关酶的活性的测定发现,喷施Men-myco-93-63发酵液后黄瓜叶片内POD,CAT,PAL,PPO等酶的活性在一定时间内有明显上升的趋势。     

miR-556-3p regulates viability,migration and invasion of endometrial cancer cells by targeting SASH1

AIM To investigate whether microRNA-556-3p （miR-556-3p） regulates the viability, migration and invasion of endometrial cancer cells by targeting SASH1 gene. METHODS The expression of miR-556-3p, and the mRNA and protein levels of SASH1 in endometrial cancer tissues were detected by RT-qPCR and Western blot. Anti-miR-556-3p or pcDNA-SASH1 was transfected into endometrial cancer Ishikawa cells. The cell viability was detected by MTT assay, the migration and invasion abilities of the cells were detected by Transwell chamber method, and the protein expression levels of cyclin D1, p21, matrix metalloproteinase-2 （MMP-2） and MMP-9 were detected by Western blot. StarBase prediction and dual-luciferase reporter experiments were used to analyze the targeting relationship between miR-556-3p and SASH1. Anti-miR-556-3p and si-SASH1 were co-transfected into the Ishikawa cells, and their effects on cell viability, migration and invasion were examined by the methods described above. RESULTS Compared with adjacent tissues, the expression of miR-556-3p in endometrial cancer tissues was increased significantly, and the expression of SASH1 at mRNA and protein levels was decreased significantly （P<0.05）. Inhibition of miR-556-3p expression or induction of SASH1 over-expression obviously reduced the viability of Ishikawa cells, the number of migratory cells, the number of invasive cells and the protein levels of cyclin D1, MMP-2 and MMP-9, and dramatically increased the protein level of p21 （P<0.05）. miR-556-3p targeted SASH1 and negatively regulated its expression. Knock-down of SASH1 expression reversed the inhibitory effect of miR-556-3p expression inhibition on the viability, migration and invasion of Ishikawa cells. CONCLUSION Inhibition of miR-556-3p expression suppresses the viability, migration and invasion of endometrial cancer cells. The mechanism is related to the regulation of its target gene SASH1.     

鸡新城疫La Sota株、鸡传染性支气管炎W93株二联活疫苗研究Ⅱ.接毒剂量选择试验

将鸡新城疫La Sota毒种和嗜肾型鸡传染性支气管炎W93毒种分别用生理盐水做适当稀释后，将两种病毒液等量混合，接种于同一SPF鸡胚尿囊腔内。结果表明，HBV、NDV在同一鸡胚内增殖时无互相干扰现象；收获的鸡胚液(La Sota W93)中NDV、IBV的效价分别达到相应单苗水平。     

大豆新品种科选93选育与应用

大豆新品种科选93系唐山市农业科学研究所选育而成.该品种具有高产、早熟、矮秆、分枝多、抗花叶病毒病的特点.可作为1年2熟或2年3熟制地区夏播品种和2年3熟及1年1熟制地区春播品种.于2002年4月通过河北省农作物品种审定委员会审定.     

MicroRNA-24 regulates apoptosis of cardiomyocytes after myocardial infarction

AIM：To evaluate the expression of miR-24 in infarcted myocardial tissues and to investigate the function of miR-24 during cardiomyocyte apoptosis in vitro and in vivo. METHODS：The mouse model of myocardial infarcton (MI) was established. The expression of miR-24 in the sections of infarcted myocardial tissues was measured by qRT-PCR. The expression of miR-24 was modified by transfecting oligonucleotide mimic and inhibitor of miR-24 into cardiomyocytes, or injecting lentiviral vectors intramyocardially. The apoptosis of cardiomyocytes was detected by Caspase-Glo 3/7 Assay System. The heart functions were determined by echocardiography and the scar size in MI model was observed with Masson trichrome staining. The apoptosis of infarcted myocardial tissues was detected by TUNEL method. Microarray and bioinformatic analysis were also used to predict the targets of miR-24. RESULTS：The expression of miR-24 in the infarct and border areas was down-regulated after MI. Overexpression of miR-24 in cardiomyocytes reduced the apoptosis induced by hypoxia. miR-24 transfection resulted in reduction of the scar size, and improved left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) of the mice in treatment group after 2 weeks. Furthermore, miR-24 reduced cell apoptosis in the infarct region. BCL2L11, ANK3 and SGPL1 may be the targets of miR-24 during the process of anti-apoptosis. CONCLUSION：miR-24 is down-regulated in the infarcted myocardial tissues. In vitro, miR-24 reduces apoptosis of cardiomyocytes induced by hypoxia. In vivo, miR-24 attenuates cell apoptosis in the infarct and border areas of the heart 2 weeks after MI, and ultimately improves heart functions.     

高温条件下miRNA-24对奶牛乳腺上皮细胞增殖与凋亡的影响

【目的】MicroRNA(miRNA)是一类长度约21-23个核苷酸的非编码小RNA分子,本研究旨在分析高温条件下miRNA-24对乳腺上皮细胞生长、凋亡的作用及其初步机制。【方法】体外培养奶牛乳腺上皮细胞,高温(41℃)处理所培养的细胞;应用miRNA基因沉默技术,抑制高温处理过的乳腺上皮细胞中miRNA-24的表达;采用细胞计数法分析细胞增殖情况;Hoechst33342/PI荧光染色、流式细胞术检测miRNA-24对乳腺上皮细胞凋亡的作用,Western blot检测凋亡相关因子表达变化。【结果】抑制miRNA-24的表达,高温条件下的乳腺上皮细胞增殖能力显著增强(P0.05),凋亡细胞数显著减少(P0.05),促凋亡因子caspases-8和caspases-3表达量下调。【结论】内源性miRNA-24对乳腺组织发育具有重要的调控作用,抑制miRNA-24的表达可以缓解高温诱导下奶牛乳腺上皮细胞凋亡发生率、促进细胞的生长发育。