玉米粉中伏马毒素FB1基体标准物质的评价

为填补我国在伏马毒素基体标准物质的空白,本试验研制了以玉米粉为基体的伏马毒素FB1标准物质,玉米样品经过筛、加标、冷冻干燥、磨粉、混匀、密封分装后,用超高效液相色谱-串联质谱(UPLC-MS/MS)法检验样品的均匀性、稳定性,幵联合多家实验室对玉米粉中伏马毒素FB1含量定值,同时分析样品的不确定度。样品均匀性经斱差分析法表明F值为1.42,小于临界值F_(0.05),且伏马毒素FB1含量在觃定时间6个月内无明显变化。结果表明,均匀性与稳定性均符合标准物质的要求。样品定值为1475.56μg kg~(–1),不确定度为169.98μg kg~(–1)。该标准物质可替代进口标准物质,用于伏马毒素检测过程中的仪器校准、实验室质量控制和操作人员的水平考核等。     

SCFAs对奶牛瘤胃上皮细胞Ca2+信号通路相关基因表达的影响

为探究短链脂肪酸(SCFAs)对奶牛瘤胃上皮细胞(BRECs)Ca~(2+)信号通路相关基因表达的影响,试验分为3个处理组,分别为野生型BRECs组,含20mmol/L SCFAs BRECs组,含20mmol/L SCFAs且通过CRISPR/Cas 9系统敲除GPR41基因的BRECs组,每个处理组3个重复,每组细胞均培养24h后,收集细胞提取总RNA,通过qRT-PCR对Ca~(2+)信号通路相关基因的mRNA表达量和细胞内Ca~(2+)浓度进行测定。结果表明,与野生型BRECs相比,添加20mmol/L SCFAs可极显著增加PLCB2的mRNA表达量(P0.01),显著增加IP3R1的mRNA表达量(P0.05),对PLCE1、PLCL1、PKCB和PKCG的mRNA表达量无显著差异(P0.05),可增加细胞内Ca~(2+)浓度但无显著差异(P0.05);敲除SCFAs的受体GPR41后,添加20 mmol/L SCFAs可显著降低IP3R1的mRNA表达量(P0.05),极显著上调PLCE1和PLCB2的mRNA表达量(P0.01),但对PLCL1、PKCB和PKCG的mRNA表达量无显著影响(P0.05),细胞内Ca~(2+)浓度有降低趋势但无显著差异(P0.05)。综上,SCFAs可以通过激活其受体GPR41来调控BRECs内Ca~(2+)信号通路中相关基因的表达和细胞内Ca~(2+)的释放。     

秀珍菇原基形成相关基因PpFBD1的克隆与表达研究

前期研究中通过对秀珍菇经低温诱导后不同发育阶段样本进行转录组测序及差异表达基因分析发现,ID为Cluster-6377.64510的基因(命名为PpFBD1)在原基形成前期的样本中高表达。为进一步了解分析其表达特性及功能,根据转录组测序结果设计特异性引物,利用RT-PCR技术从秀珍菇中克隆获得了该基因的cDNA全长。该基因由342个核苷酸组成,编码一个由113个氨基酸组成的蛋白,蛋白相对分子质量约11 ku。进化树分析显示,PpFBD1编码蛋白与糙皮侧耳hydrophobin1编码的疏水蛋白亲缘关系最近。Gene Ontology功能分析表明,PpFBD1主要参与真菌类细胞壁的合成。荧光定量RT-PCR检测显示,该基因在菌丝体经低温诱导后恢复至室温阶段(原基形成前期)表达量最高,这表明其可能参与调控秀珍菇原基形成过程。本研究结果为阐明秀珍菇原基形成机制提供参考。     

Effect of HMGA2 gene on high glucose-induced apoptosis of renal tubular epithelial cells by regulating Notch signaling pathway

AIM:To investigate the effect of HMGA2 down-regulation on apoptosis and Notch signaling pathway in renal tubular epithelial cells exposed to high glucose (HG). METHODS:D-glucose at 5, 10, 20 and 30 mmol/L was used to stimulate human renal tubular epithelial HK-2 cells for 2 h, and D-glucose at 30 mmol/L was used to stimulate the HK-2 cells for 10 min, 60 min and 120 min. The protein expression of HMGA2 was determined by Western blot. The HK-2 cells were divided into normal glucose (NG) group, HG group, HG+si-HMGA2 group and HG+NC group, in which siRNA was transfected by Lipofectamine^TM 2000 for 48 h. Flow cytometry was used to analyze the apoptotic rate, reactive oxygen species (ROS) assay kit was used to detect ROS content, and Western blot was used to detect the protein levels of Notch1, Hes1 and Bcl-2. The HK-2 cells were treated with the Notch signaling pathway inhibitor DAPT, and then the cells were divided into HG group, HG+DAPT group and HG+si-HMGA2+DAPT group. The apoptotic rate was analyzed by flow cytometry. RESULTS:Exposure of the HK-2 cells to D-glucose at different concentrations for different time significantly increased the expression of HMGA2 (P<0.05). Compared with NG group, the protein expression of HMGA2, Notch1 and Hes1 in HG group was increased, the expression of Bcl-2/Bax was decreased, the apoptotic rate was increased, and the content of ROS was increased obviously (P<0.05). Compared with HG group, the protein expression of HMGA2, Notch1 and Hes1 of HG+si-HMGA2 group was decreased, the expression of Bcl-2/Bax was increased, the apoptotic rate was decreased, and the content of ROS was decreased significantly (P<0.05). The apoptotic rate in HG+DAPT group was significantly lower than that in HG group, while the apoptotic rate in HG+si-HMGA2+DAPT group was significantly lower than that in HG+DAPT group (P<0.05). CONCLUSION:Down-regulation of HMGA2 expression inhibits the apoptosis of renal tubular epithelial cells by regulating Notch signaling pathway and decreasing ROS production.     

不同贮藏温度和保鲜处理对黄秋葵贮藏品质及生理的影响

为探讨采后不同贮藏温度对黄秋葵贮藏品质的影响以及不同保鲜剂对黄秋葵抗氧化酶活性的影响，以台湾“五福”黄秋葵为试材，分别置于低温（（4±1）℃、（9±1）℃、（14±1）℃）和常温（（20±1）℃）条件下贮藏，研究温度对其贮藏品质的影响；采用1.0%壳聚糖、5.0 mmol/L水杨酸、0.5片安喜布、5%O2+8%CO2处理黄秋葵，研究保鲜处理对其抗氧化酶活力的影响。结果表明：（9±1）℃贮藏组的保鲜效果最好，显著改善了贮藏期间黄秋葵的失重率、硬度、叶绿素含量、VC含量、L值、呼吸强度、乙烯释放量、MDA含量、细胞膜相对电导率；4种保鲜处理均能显著提高黄秋葵的SOD、POD、CAT活性，并维持其活性在相对较高的水平；同时，显著抑制黄秋葵果实MDA含量的升高。通过研究发现，壳聚糖涂膜液处理组的效果最佳，水杨酸处理组次之并优于1-MCP处理组，气调处理组的效果相对较差。     

Selective inhibition of endothelin receptor A improves white matter lesions in hypertensive rats

AIM:To observe whether selective inhibition of endothelin receptor A (ETRA) improves white matter lesions (WMLs), and explore the mechanism. METHODS:Sprague-Dawley rats (n=33) were randomly divided into sham operation group (n=9), treatment group[stroke-prone renovascular hypertensive rats-modified 2 vessel occlusion (RHRSP-modified 2VO) + ambrisentan (n=12)] and placebo group[RHRSP-modified 2VO + vehicle (n=12)]. Drug and vehicle administration was performed from 17th to 20th week and monitoring of systolic arterial pressure was performed weekly. Morris water maze test was conducted to evaluate the function of cognition. The protein levels of endothelin-1 (ET-1) in the cortex, corpus callosum and caudate putamen were quantitatively analyzed respectively. The severity of WMLs and the relationship between ET-1 and vessels were observed by the method of histopathology. RESULTS:The difference of systolic arterial pressure between treatment group and placebo group was not significant. The animals in treatment group exhibited shorter escape latency (P<0.05), more times of crossing platform (P<0.05), lower level of ET-1 in corpus callosum and caudate putamen (P<0.05), respectively, improved WMLs severity (P<0.05) and lower binding level of ET-1 to vessels compared with the placebo group. CONCLUSION:Selective inhibition of endothelin receptor A improves the severity of WMLs and ameliorates the cognitive function.     

Effects of taurine-zinc on learning and memory abilities of vascular dementia mice

AIM:To observe the effects of taurine-zinc (TZC) on the learning and memory abilities of vascular dementia (VD) mice and to investigate the related mechanism. METHODS:The mice were randomly divided into model group, sham group, and TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg groups. The mice in drug groups were given TZC by gavage at 10 mL/kg once daily. The mice in sham group and model group were given equal volume of distilled water. VD mice were established by intercepting both common carotid arteries and bleeding at caudal vein after 14 d of gavage. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISA. The levels of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) were measured via spectrophotometer. Step-down test and Morris water maze test were used to examine the abilities of learning and memory in the mice. RESULTS:TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg reduced the levels of TNF-α, IL-1β, iNOS and NO in the brain tissues. In the water maze test, TZC at 100 mg/kg and 200 mg/kg significantly decreased the error times and latency compared with model group. In the step-down test, the escape latency was prolonged and error times were lowered significantly by treatment with TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg as compared with model group. CONCLUSION:TZC improves the abilities of learning and memory, which might be related to the reduction of TNF-α, IL-1β, iNOS and NO levels in VD mice.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.