不同处理褐煤腐殖酸对磷吸附特性的研究

采用吸附等温线的试验方法,研究了平衡溶液中磷浓度、反应时间及介质pH对不同处理的2种褐煤腐殖酸吸附磷的影响及其规律性。结果表明:在试验pH范围内,pH的升高减缓了吸附反应进行的速度,对于未经硝酸处理的样品S1,在pH为4.7时,吸附量和分配系数有一最大值,而样品S2,随着pH的升高,吸附量和分配系数一直减小;经硝酸处理后,样品对磷的吸附能力降低,一定条件下其单位吸附量仅为处理前的5.33%。适宜的固液比例可提高2个不同处理供试样品对磷的单位吸附量,其等温吸附规律可用Langmuir方程来加以描述;其吸附动力学用Elovich方程描述最佳。     

绵羊甲状腺miR-370-3p靶向COL4A3基因调控繁殖力性状的初步研究

为探讨小尾寒羊甲状腺组织中与繁殖功能相关的miR-370-3p与COL4A3的靶向关系,本研究利用miRDB与Targetscan网站预测了miR-370-3p的靶基因,并取其交集进行功能富集分析,通过RNAhybrid与Targetscan网站预测绵羊miR-370-3p与候选靶基因的结合位点;随后检测了miR-370-3p与COL4A3基因在小尾寒羊甲状腺组织中的相对表达量。构建psiCHECK2-COL4A3-3′UTR-野生型/突变型双荧光素酶载体,并将其与miR-370-3p mimics/mimics NC共转染至HEK293T细胞并检测荧光活性。结果显示：1)miR-370-3p的靶基因可富集到孕酮介导的卵母细胞成熟、卵母细胞减数分裂、HIF-1信号通路、MAPK信号通路、mTOR信号通路、Ras信号通路和FoxO信号通路等与动物繁殖有关的信号通路中。2)miR-370-3p可与COL4A3基因3′UTR区域结合,且两者在小尾寒羊甲状腺组织中表达呈相反趋势。3)共转染psiCHECK2-COL4A3-3′UTR-野生型和miR-370-3p mimics组的荧光素酶活性显...     

‘72杨’韧皮部的构造与理化性能

将‘72杨’、杉木和毛竹的木质部进行对比试验,观察并测试‘72杨’韧皮部的解剖构造以及理化特性,为其高值化利用提供基础数据。使用场发射环境扫描电镜、X射线衍射仪、傅里叶变换红外光谱仪等设备和NREL标准对‘72杨’韧皮部的微观结构、结晶度、化学成分等物化性质进行测定与分析。研究结果表明,‘72杨’韧皮部中韧皮薄壁细胞和筛管分子占细胞总面积的(81.9±1.8)%,结构相对简单。‘72杨’韧皮部气干密度为0.358 g/cm3,绝干密度为0.321 g/cm3;而木质部的相应密度较高,分别为0.497和0.482 g/cm3;‘72杨’韧皮部结晶度仅为19.4%,比木质部低8.7%。‘72杨’韧皮部纤维素、半纤维素、木质素的含量分别为28.7%,11.1%,24.1%,均低于木质部中相应成分的含量,且木质化程度低,半纤维素以木糖为主。此外,由红外谱图发现‘72杨’韧皮部含有单宁、酚类、胼胝质等物质。‘72杨’韧皮部具有低密多孔、结构疏松、结晶度低、木质素含量低、抽提物含量高等特点。因此,‘72杨’韧皮部特别有利于机械(能耗低)或化学(抗降解屏障低)降解以及物化改性(多孔、可及性强),可提取酚类、单宁、胼胝质等物质用于工业应用,研究结果可为‘72杨’韧皮部的高值化利用提供重要的理论依据。     

Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

基于VP72基因的非洲猪瘟PCR检测方法的建立与应用

基层兽医快速准确诊断非洲猪瘟,可以加强地方兽医部门的疫情防控能力。笔者以非洲猪瘟结构基因VP72部分序列为检测靶基因,设计特异性引物,优化反应条件,建立常规PCR检测方法。笔者应用新建的检测方法,检测了贵州境内采集的5份病料,其中4份阴性,1份阳性,与非洲猪瘟荧光实时定量PCR复检结果一致。结果表明,笔者新建的PCR检测方法特异性好,与其他病原无交叉性,快捷便利。同时也表明,贵州非洲猪瘟疫情暴发压力较大,应加强疫情防范和扑灭工作。     

Outer membrane protein A of Acinetobacter baumannii induces autophagy via Akt/mTOR/p70S6K signaling pathway in RAW264.7 cells

AIM:To analyze the effects of outer membrane protein A (OmpA) from Acinetobacter baumannii ATCC 19606 on the autophagy of RAW264.7 cells. METHODS:The RAW264.7 cell model stimulated by OmpA was established. The effects of OmpA on the autophagy of RAW264.7 cells were detected by immunofluorescence, Western blot and transmission electron microscopy. RESULTS:The OmpA increased the expression of LC3B-Ⅱ and reduced the phosphorylation levels of Akt, mTOR and p70S6K. Rapamycin further reduced the phosphorylation levels of mTOR and p-70S6K, and increased the expression of LC3B-Ⅱ induced by OmpA. CONCLUSION:The OmpA of Acinetobacter baumannii induces autophagy via Akt/mTOR/p70S6K signaling pathway in the RAW264.7 cells. This work provides a basis for further research on the molecular mechanism of autophagy induced by Acinetobacter baumannii to find a new method against the infection of Acinetobacter baumannii.     

microRNA-124-3p调控H1N1亚型猪流感病毒致小鼠肺损伤的研究

为研究microRNA-124-3p（miR-124-3p）对H1N1亚型猪流感病毒（swine influenza virus，SIV）感染小鼠所致肺损伤的调控作用，本试验构建miR-124-3p腺病毒表达载体，通过小鼠尾部静脉注射法构建miR-124-3p差异表达小鼠模型，试验分3组：过表达组、抑制组和对照组。48 h后，各组小鼠鼻腔接种H1N1亚型SIV，每只10⁵ EID₅₀（50 μL）。连续观察14 d，计算小鼠平均体重变化率、观察病理切片并测定相关炎症因子IL-1β、TNF-α和IL-6 mRNA相对表达量。结果显示，已成功将pre-miR序列及其sponge序列插入腺病毒的穿梭质粒，并将其共转染293A细胞。实时荧光定量PCR检测证实，与对照组相比，过表达组和抑制组小鼠黑色素瘤细胞miR-124-3p表达水平分别极显著升高（P<0.01）和显著降低（P<0.05），表明成功构建腺病毒表达载体。过表达组、抑制组和对照组小鼠体重变化率分别为－5.5%、－12.4%和－8.6%。抑制组和对照组均可见肺泡壁增厚，其间有多量淋巴细胞浸润，部分肺泡内出现纤维蛋白渗出，且抑制组病理变化更为严重，肺泡中还有大量的红细胞浸润；而过表达组仅有少量的淋巴细胞浸润，肺脏组织较正常。与对照组相比，过表达组检测的炎症因子IL-1β、TNF-α和IL-6 mRNA表达水平均显著降低（P<0.05）；抑制组炎症相关炎症因子mRNA表达水平均显著升高（P<0.05）。本试验结果表明，miR-124-3p对H1N1亚型SIV感染小鼠所致的肺脏炎症因子的表达具有抑制作用，同时能减轻肺脏病理损伤。     

广谱驱虫药帕苯达唑的合成

以对正丁基苯胺为原料,经乙酰化、硝化、水解、还原、闭环反应合成了5-正丁基-1H-苯并咪唑-2-氨基甲酸甲酯（帕苯达唑）。首先将乙酰化和硝化采用“一锅法”得到4-正丁基-2-硝基乙酰苯胺,再以甲醇作溶剂,碱性水解后直接以钯碳为催化剂,在40℃、0.5MPa条件下反应得到4-正丁基邻苯二胺。最后选用氰氨基甲酸甲酯为闭环剂,制得目标产物。整个工艺流程简化了工艺条件,降低了生产成本,总收率可达41.8%,适合工业化生产。     

Involvement of the ERK1/2 MAPK pathway in insulin-induced S6K1 activation in avian cells

In mammals, insulin regulates S6K1, a key enzyme involved in the control of protein synthesis, via the well-documented phosphoinositide-3'kinase (PI3K) pathway. Conversely, S6K1 is activated by insulin in avian muscle despite the relative insulin insensitivity of the PI3K pathway in this tissue. Mitogen-activated protein kinase (MAPK) cascade is another insulin sensitive pathway. The aim of this study was to explore the potential involvement of the ERK1/2 MAPK pathway in the control of p70 S6 kinase (S6K1) in avian species. Firstly, we characterized ERK1/2 MAPK in various chicken tissues. ERK2 was the only isoform detected in avian species whatever the tissue studied. We also showed that ERK2 is activated in vivo by insulin in chicken muscle. The regulation and the role of ERK2 in insulin signaling were next investigated in chicken hepatoma cells (LMH) and primary myoblasts. Insulin stimulation led to ERK2 and S6K1 phosphorylation, and concomitantly increased kinase activity. U0126, an inhibitor of the ERK MAPK pathway, completely abolished insulin-induced S6K1 phosphorylation and activity in chicken myoblasts, whereas its effect was only partial in LMH cells. In conclusion, these results show that ERK1/2 MAPK is involved in the control of S6K1 by insulin in chicken cells, particularly myoblasts.     

Radiosensitivity of canine osteosarcoma cells transfected with wild‐type p53 in vitro*

The p53 gene is one of the important tumour suppressor genes that are involved with the cell survival signal pathway. One of the major functions of the p53 protein is to organize cell cycle regulation and induction of apoptosis for cellular genetic stability. It has been documented that more than 50% of all human cancers include a p53 mutation. We evaluated the difference in radiosensitivity between upregulating the expression of canine wild‐type p53 (cp53) in cultured osteosarcoma (D17) cells and naive D17 cells in vitro. We found that upregulating transfected cp53 D17 cells increased their radiation sensitivity in vitro, and there was a significant decrease (P < 0.009) in survival between cp53‐transfected D17 cells and naive D17 cells. In this experiment, a p53 enhancement ratio (p53ER) reached approximately 3.0 at high doses. The transfected cp53 D17 cells were significantly more radiosensitive at all doses evaluated than naive D17 cells, except at 1 Gy where too few data points were available. The p53ER increased rapidly at doses less than 4 Gy, achieving a maximum of about 3.0 for doses of 4 Gy and above. This study shows the enhanced radiosensitivity of the transfected p53 at clinically relevant doses.