Calreticulin promotes migration and invasion of nasopharyngeal carcinoma cells by inducing cell EMT

AIM:To observe the expression of calreticulin (CRT) in nasopharyngeal carcinoma tissues, analyze the significance of clinical pathology and the influence on epithelial-mesencymal transition (EMT) of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues, and the significance of clinical pathology was evaluated. The calreticulin gene-specific small interfering RNA was constructed, and then was transfected into the NPC cell line CNE2 using the cationic liposome method. The effect of CRT on the morphological changes of the CNE2 cells was observed under light microscope. The effect of CRT on the cell migration and invasion abilities of the CNE2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin, vimentin, transforming growth factor (TGF)-β and matrix metalloproteinase (MMP)-9 in the CNE2 cells was determined by Western blot. RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29% (11/57), which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69% (43/52). The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis (P<0.05). Knockdown of CRT expression made the CNE2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change, arranged more compact, and the migration and invasion abilities were significantly decreased (P<0.05). Knockdown of CRT expression resulted in significant increase in the protein expression of E-cadhe-rin, and the decreases in the protein expression of vimentin, TGF-β and MMP-9 in the CNE2 cells (P<0.05). CONCLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with nasopharyngeal carcinoma stage and lymph node metastasis. Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.     

60Co辐射诱变姬松茸突变株J3子实体不同部位的氨基酸分析研究

测定并比较了60Co辐射诱变姬松茸突变株J3与原菌株J1子实体不同部位的氨基酸含量.结果表明,除菌柄和盖皮外,姬松茸突变株J3子实体其它部位的必需氨基酸、儿童氨基酸、硫氨酸、支链氨基酸、鲜味氨基酸、芳香族氨基酸和甜味氨基酸均高于原菌株J1.姬松茸突变株J3子实体中谷氨酸含量占氨基酸总量的比例分别比原菌株J1、α-酪蛋白和卵清蛋白提高34.88%、56.68%和122.32%.     

禽脑脊髓炎病毒Van Roekel株衣壳蛋白基因原核表达及其ELISA检测研究

利用RT PCR方法扩增了禽脑脊髓炎病毒VP0、VP3和VP1基因 ,分别插入pGEX 4T 1原核表达载体 ,转化到E .coliER2 566表达菌内 ,经IPTG诱导表达后 ,SDS PAGE分析表达产物。结果表明 ,表达的VP0、VP3、VP1融合蛋白分子量分别为 53 ,53 ,56ku。以电洗脱法纯化VP0、VP3、VP1表达蛋白抗原并分别建立了间接ELISA方法 ,用阳性血清和阴性血清进行检测。结果显示 ,表达VP1蛋白反应活性相对高于VP0、VP3蛋白反应活性 ,最后再将自制VP1板用自配试剂检测其活性 ,同时用美国试剂盒检测VP1活性 ,美国ELISA试剂盒、琼脂扩散试验检测同样 1 0份血清作比较 ,三组ELISA检测结果 ,其中有 8份被检血清为阳性 ;2份被检血清为阴性。这一结果与琼脂扩散试验检测结果一致。说明VP1表达蛋白活性达到预期要求 ,可以用做AE的ELISA诊断     

毛茛科植物提取物添加量对瘤胃微生物体外动态发酵的影响

试验采用活体外产气量法研究毛茛科植物提取物不同添加水平对瘤胃微生物体外发酵的影响。试验采用单因子完全随机化试验设计,以小麦面粉为底物进行体外发酵。瘤胃液供体动物为3头装有永久性瘤胃瘘管的本地黄牛,日粮精料水平为30%(日饲喂2次)。植物提取物添加水平分别为0、100、200、300和400mg/L。结果表明:随着毛茛科植物提取物添加水平的提高,小麦面粉24h的DM消化率呈线性下降(P=0.0002),活体外产气速度呈二次曲线规律降低(P=0.0001),而产气延滞期线性增加(P=0.0001),理论最大产气量呈二次曲线规律增加(P=0.0001)。随着植物提取物添加水平的提高,发酵液pH值呈二次曲线增加(P<0.024)。各培养时间点乳酸含量均较低(P<1mmol/L),且各处理之间差异不显著(P>0.10)。提高植物提取物添加水平导致tVFA产量呈二次曲线趋势下降(P<0.01)。通过本试验可得出:毛茛科植物提取物可以通过降低瘤胃发酵的产气速度、提高发酵液pH值以及乙酸和丁酸摩尔比例来调控瘤胃微生物体外发酵,其中植物提取物的适宜添加水平为200～300mg/L。     

蚕粉复合物对小鼠若干生理指标的影响

以小鼠为试验动物 ,对以全蚕粉为主要成分的降血糖复合物 (SPC)的生理作用和药效进行了探讨。结果表明 ,该复合物可以显著降低小鼠的食后血糖 ,并能提高小鼠抗疲劳能力和耐力 ,对小鼠的生长发育、繁殖等无不良影响 ,未发现有任何毒副作用。     

柑桔黄龙病菌在贡柑叶片中的分布分析

2017年11月在广东省仁化县4个感染柑桔黄龙病的贡柑园中采叶片样品,进行常规PCR和实时荧光定量PCR检测,从不同病症叶片、典型症状叶片不同部位、初期感病树不同部位(正常和显症)叶片等3个方面分析了贡柑叶片中黄龙病菌"Candidatus Liberibacter asiaticus"(CLas)的分布情况。结果表明,叶片斑驳黄化症和均匀黄化症是与黄龙病最相关的症状,CLas含量很高;缺锌黄化症,具有黄龙病阳性与阴性两类样品;无明显黄化但呈革质化的叶片,CLas含量高。在斑驳黄化与均匀黄化叶片中,CLas的含量从叶尖到叶柄、从叶片到主叶脉呈现递增趋势,即以主叶脉与叶柄的含量更高。初期感病树未显症的部位也会检出黄龙病菌。     

设施韭菜安全生产关键技术集成

生产绿色、安全、放心韭菜，除了应用优质品种、实施绿色韭菜安全施肥技术外，推广绿色韭菜韭蛆防控新技术是关键。     

Establishment of a Duplex Real-time RT-PCR Assay for the Differential Detection of Nipha Virus and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus

To establish a rapid,sensitive and specific assay for the differential detection of Nipah virus (NiV) and highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV),a duplex Real-time RT-PCR was developed with specific primers and probes targeting to the special sequences of NiV M gene and HP-PRRSV nsp2 gene by optimization of reaction conditions.The performance of the assay was linear ranging from 4.6×10¹ to 4.6×10⁷ copies/μL for RNA standard control of NiV M (NiV-M-RNA) and from 4.1×10¹ to 4.1×10⁸ copies/μL for RNA standard control of HP-PRRSV nsp2 (HP-PRRSV-nsp2-RNA),and detection limits of the assay was 46 copies for the NiV-M-RNA and 4.1 copies for the HP-PRRSV-nsp2-RNA,respectively.The coefficients of variation (CVs) of both inter-assay and intra-assay repeatability were less than 2.0%,showing good repeatability.The assay was able to specifically detect NiV and HP-PRRSV simultaneously without cross-reaction with classical swine fever virus (CSFV),porcine epidemic diarrhea virus (PEDV),swine influenza virus (SIV),porcine parvovirus (PPV),pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2).Of the 236 samples from pigs for both NiV and HP-PRRSV detection by the established assay,all the samples were negative for NiV,8 samples were HP-PRRSV positive.In conclusion,this assay offers a useful approach for the differential detection of NiV and HP-PRRSV in clinical specimens from the pigs.     

苹果果实表皮总蛋白提取及双向电泳方法的优化

以苹果果实表皮为试材,采用3种方法提取果实表皮总蛋白,并对双向电泳(2-DE)操作体系进行优化。结果表明:改良酚提法适于苹果果实表皮总蛋白的提取,优化后的2-DE分离体系为预制IPG胶条18cm pH 4~7,上样量600μg,改进后的等电聚焦程序,浓度16%的分离胶,适于苹果果实表皮总蛋白的2-DE分离。     

天门市蔬菜产业发展现状及对策

分析了天门蔬菜产业发展现状,指出其发展过程中存在的问题,提出了加大基地配套设施建设力度、优化蔬菜品种结构、促进集约规模化生产、提高产品市场竞争力、完善质量安全保障体系、强化宣传监管、构建蔬菜市场流通体系的解决对策,以期为天门蔬菜产业的健康持续发展提供参考。