硒对硒蛋白-谷胱甘肽过氧化物酶基因表达及其酶活性的调节

本文根据国内外研究现状详细综述了硒调节细胞型谷胱甘肽过氧化物酶(GPX1)mRNA水平、蛋白水平及酶活3个水平的可能分子机理,初步概述硒对其余GPX基因表达的影响及其作用机制,为深入研究硒调节GPX基因表达的作用机制提供参考。     

大豆疫霉菌对大豆下胚轴侵染过程的细胞学研究

 接种后1.5~24h,用光镜和电镜研究了2个大豆品种与大豆疫霉菌Ps411的亲和性和非亲和性互作。观察结果表明,大豆疫霉菌对大豆下胚轴的侵染过程可分为侵入前、侵入、皮层组织中的扩展和进入维管束组织4个连续阶段。大豆下胚轴接种后在25℃保湿培养,1.5h后游动孢子即形成休止孢并萌发产生附着孢,3h后侵入表皮细胞,6h后进入皮层组织,24h后进入维管束组织。病原菌主要以侵染菌丝直接侵入表皮,表皮细胞间隙是主要侵入部位。皮层细胞是病原菌定殖和发展的主要场所,胞间菌丝侵入皮层细胞并形成吸器。在菌丝与寄主细胞接触部位的寄主细胞壁与质膜之间常有胞壁沉积物的形成。在抗病品种上病菌的侵染事件与感病品种基本一致,但不能形成正常的吸器,胞壁沉积物明显多于感病品种,菌丝在寄主组织内的扩展明显受到抑制。利用β-1,3-葡聚糖免疫金标记单克隆抗体进行的免疫细胞化学的研究表明,胞壁沉积物内含有大量的β-1,3-葡聚糖,在大豆疫霉菌菌丝壁中也存在β-1,3-葡聚糖。以上结果表明,病原菌的侵染可诱导抗病寄主细胞内β-1,3-葡聚糖迅速的合成与积累、并形成胞壁沉积物,以抵御病菌的侵染与扩展。     

Irbesartan alleviates hepatic steatosis in db/db mice by inducing auto-phagy

AIM: To investigate the effect of irbesartan on the fatty liver of db/db mice and whether autophagy is involved in the process. METHODS: Male db/db mice (n=24) were randomly divided into model group and irbesartan group, and 12 db/m mice with similar age and weight were selected as normal control group. After 16 weeks of intervention respectively, the fatty liver-related parameters including body weight, liver index, blood lipid, liver function and pathological changes in the liver were observed. The protein levels of p-PI3K, p-Akt, and p-mTOR, as well as Atg-7, beclin-1 and LC3B in the liver tissues were detected by Western blot, and the autophagosomes in the liver were observed under electron microscope. RESULTS: Compared with the model group, the body weight, liver index, blood lipids, alanine and aspartate aminotransferase were decreased in irbesartan group (P<0.05). Moreover, the pathological changes in the liver were significantly ameliorated in irbesartan group than that of model group. Importantly, the protein levels of p-PI3K, p-Akt and p-mTOR were decreased with irbesartan administration, while the expression of Atg-7, beclin-1 and LC3B-Ⅱ was increased(P<0.05), which resulted in a distinct increase in autophagosomes. CONCLUSION: Irbesartan alleviates hepatic steatosis in db/db mice by inhibiting the PI3K/Akt/mTOR signaling pathway and upregulating the protein expression of Atg-7, beclin-1 and LC3B-Ⅱ, thereby inducing autophagy in hepatocytes.     

饲粮不同NFC/NDF对肉用绵羊瘤胃pH、氨态氮和挥发性脂肪酸的影响

本试验旨在研究饲粮不同非纤维性碳水化合物/中性洗涤纤维(NFC/NDF)对肉用绵羊瘤胃pH、氨态氮和挥发性脂肪酸的影响.选用(47.21±1.01)kg安装有瘤胃瘘管的杜泊羊(♂)×小尾寒羊(♀)杂交1代肉用公羊12只,采用12×4不完全拉丁方设计,试验分4期进行,每期16 d,分别饲喂NFC/NDF为0.25、0.34、0.36、0.52、0.60、0.80、0.87、1.13、1.30、1.58、2.17和2.49的12种饲粮.结果表明:随着NFC/NDF的增加,试验羊瘤胃pH极显著线性降低(P＜0.01),氨态氮浓度极显著线性增加(P＜0.01),瘤胃总挥发性脂肪酸及丁酸比例呈显著三次曲线变化(P＜0.05),总挥发性脂肪酸中的丙酸、戊酸和异戊酸比例极显著线性增加(P＜0.01),乙酸比例和乙酸/丙酸极显著线性降低(P＜0.01).由此可见,饲粮NFC/NDF对瘤胃pH、氨态氮和挥发性脂肪酸具有显著影响.     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Down-regulation of miR-153-3p attenuates H2O2-induced H9C2 cell injury by promoting Nrf2 expression

AIM To study the effect of microRNA-153-3p (miR-153-3p) knock-down on oxidative injury of H9C2 cells induced by H₂O₂ and its specific mechanism. METHODS The oxidative stress injury of H9C2 cell model was induced by H₂O₂, and then the cell viability and the expression of miR-153-3p were detected by MTT assay and RT-qPCR, respectively. The effects of miR-153-3p knock-down on the H9C2 cell injury under oxidative stress were studied by RNA interference technology. The targets of miR-153-3p were identified by Western blot and dual-luciferase reporter assay. RESULTS MTT assay showed that the viability of H9C2 cells was decreased with the increase in H₂O₂ concentration (P<0.05). The results of RT-qPCR showed that the expression of miR-153-3p was increased with the increase in H₂O₂ concentration (P<0.05). Knock-down of miR-153-3p increased the viability of H9C2 cells under oxidative stress, decreased the cell apoptosis and the content of malondialdehyde (MDA), and increased the activity of superoxide dismutase (SOD). The expression of nuclear factor E2-related factor 2（Nrf2） and antioxidant response element（ARE） activity were increased with the increase in H₂O₂ concentration (P<0.01). TargetScan analysis and dual-luciferase reporter assay showed that Nrf2 was one of the potential target genes of miR-153-3p. The results of Western blot further showed that over-expression of miR-153-3p inhibited the expression of Nrf2 (P<0.01), while down-regulation of miR-153-3p increased the expression of Nrf2 (P<0.01). Dual interference with Nrf2 and miR-153-3p significantly reduced H9C2 cell viability, promoted the apoptosis, increased MDA content, and decreased SOD activity in the presence of H₂O₂ (P<0.01). CONCLUSION Inhibition of miR-153-3p expression attenuates the injury of H9C2 cells induced by H₂O₂ through up-regulating Nrf2/ARE signaling pathway.     

Canine GM2‐Gangliosidosis Sandhoff Disease Associated with a 3‐Base Pair Deletion in the HEXB Gene

Background

GM2‐gangliosidosis is a fatal neurodegenerative lysosomal storage disease (LSD) caused by deficiency of either β‐hexosaminidase A (Hex‐A) and β‐hexosaminidase B (Hex‐B) together, or the GM2 activator protein. Clinical signs can be variable and are not pathognomonic for the specific, causal deficiency.

Objectives

To characterize the phenotype and genotype of GM2‐gangliosidosis disease in an affected dog.

Animals

One affected Shiba Inu and a clinically healthy dog.

Methods

Clinical and neurologic evaluation, brain magnetic resonance imaging (MRI), assays of lysosomal enzyme activities, and sequencing of all coding regions of HEXA, HEXB, and GM2A genes.

Results

A 14‐month‐old, female Shiba Inu presented with clinical signs resembling GM2‐gangliosidosis in humans and GM1‐gangliosidosis in the Shiba Inu. Magnetic resonance imaging (MRI) of the dog's brain indicated neurodegenerative disease, and evaluation of cerebrospinal fluid (CSF) identified storage granules in leukocytes. Lysosomal enzyme assays of plasma and leukocytes showed deficiencies of Hex‐A and Hex‐B activities in both tissues. Genetic analysis identified a homozygous, 3‐base pair deletion in the HEXB gene (c.618‐620delCCT).

Conclusions and Clinical Importance

Clinical, biochemical, and molecular features are characterized in a Shiba Inu with GM2‐gangliosidosis. The deletion of 3 adjacent base pairs in HEXB predicts the loss of a leucine residue at amino acid position 207 (p.Leu207del) supporting the hypothesis that GM2‐gangliosidosis seen in this dog is the Sandhoff type. Because GM1‐gangliosidosis also exists in this breed with almost identical clinical signs, genetic testing for both GM1‐ and GM2‐gangliosidosis should be considered to make a definitive diagnosis.     

两种盐胁迫对苦楝幼苗光合特性的影响

以一年生苦楝实生苗为试材,采用温室盆栽方式,用浓度为0%(CK)、0.2%、0.4%与0.6%的Na2SO4和Na2CO3溶液分别对幼苗进行胁迫处理21 d,比较了2种盐胁迫下幼苗光合作用和叶绿素荧光参数的变化和差异。结果表明:0.2%盐浓度对Pn、Tr、Ci、Fv/Fm、Fv/Fo、qP和NPQ没有显著影响,0.6%盐浓度则影响显著。同时,各浓度盐处理对Chl a、Chl b、Chl(a+b)、Chl a/b、Fo和Fm没有显著影响。2种盐处理对苦楝幼苗叶绿素含量没有显著影响;Na2CO3胁迫通过非气孔限制引起Pn下降,而Na2SO4引起Pn下降分别为0.2%处理时是气孔限制,0.4%和0.6%处理时是非气孔限制;幼苗光合作用和光合机构对0.2%浓度胁迫有较强耐性,对0.6%浓度胁迫耐性较弱;同时,碱性盐Na2CO3胁迫对苦楝光合作用的影响整体大于中性盐Na2SO4。     

香蕉ACC氧化酶基因(MAO3)的克隆及其表达特性分析

 根据同源扩增得到香蕉(Musa acuminata) ACC氧化酶基因(MAO3) 的核心部分, 再通过3'和5'RACE扩增上、下游序列以及Genome-Walker的方法得到启动子部分, 共获得3 718 bp长度的序列。将所得结果进行聚类分析, 发现香蕉中ACC氧化酶的氨基酸序列非常保守, 各序列间的同一性高达99% ,与单子叶植物和双子叶植物中ACC氧化酶的氨基酸序列的同源性在66.7%～71.8%之间。组织原位杂交试验表明MAO3基因的表达具有组织特异性, 初步认为是在韧皮部筛管组织中特异表达。运用实时荧光定量PCR技术, 实时监测到了MAO3基因和香蕉乙烯受体基因ERS2受机械伤诱导的定量变化。     

玉米的不同加工处理对绵羊瘤胃内pH值、NH3-N和VFA浓度的影响

选用 4头装有永久性瘤胃瘘管的内蒙古半细毛羯羊 ,饲喂 4种含有不同加工处理玉米的日粮 ,在精粗比为 30∶70 ,饲养水平为 1.3MJ日粮条件下采用 4× 4拉丁方试验设计 ,对瘤胃pH值、NH3 -N和VFA浓度进行了测定 ,并研究了它们的动态变化规律。实验结果表明 :各处理组对瘤胃内 pH值影响较小 (P >0 .0 5 ) ;不同处理组与对照组比都有降低瘤胃NH3 -N浓度的趋势 ,但差异不显著 (P >0 .0 5 ) ;对VFA浓度也有一定影响 ,但各组间差异不显著 (P >0 .0 5 )。各处理组间乙酸 /丙酸基本接近 ,均属丙酸 /乙酸发酵类型