不同浓度NAA/6BA配比对不同品种甘薯茎尖培养特性的影响

为明确不同品种甘薯茎尖培养的最佳NAA/6BA配比,设置0.1 mg/L/2.5 mg/L、0.2 mg/L/2.5 mg/L、0.2 mg/L/1.0 mg/L 3组NAA/6BA浓度配比处理,观测不同浓度激素处理对‘北京553’、‘红香蕉’、‘苏薯8号’、‘烟薯25’、‘安吉芋’、‘济徐23’、‘渝紫7号’、‘商薯19’茎尖培养成苗率、愈伤组织直径、不定根数目、叶片数和植株高度的影响,同时调查不同品种试管苗的移栽成活率。结果表明,‘红香蕉’、‘济徐23’、‘渝紫7号’、‘商薯19’在0.1 mg/L/2.5 mg/L的处理下培养效率最高;‘安吉芋’在0.2 mg/L/2.5 mg/L的处理下培养效率最高;‘苏薯8号’、‘北京553’、‘烟薯25’在0.2 mg/L/1.0 mg/L的处理下培养效率最高。因此,在甘薯茎尖培养过程中,不同甘薯品种适宜的NAA/6BA配比存在显著差异,在实际生产中应根据品种特性来进行调整激素的用量和比例。     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

保护地黄瓜新品种唐杂6 号的选育

唐杂6 号是以雌性系S16 为母本，以自交系S26 为父本配制的强雌型黄瓜一代杂种。生长势强，商品瓜短棒状， 瓜长12～14 cm，横径4.0～4.3 cm，非特异性环境下雌花率95% 以上，瓜皮嫩绿有光泽，白刺，刺瘤稀小，平均单瓜质量 131.8 g 左右，高抗霜霉病，抗细菌性角斑病，耐白粉病；春保护地栽培平均产量可达8 000 kg·（667 m²） ^-1，秋冬保护地栽 培产量可达6 500 kg·（667 m²） ^-1；适合河北、北京、天津及东北地区春、秋保护地种植。     

Expression and mechanisms of hsa_circ_087631 in patients with primary biliary cholangitis

AIM To explore the expression and mechanisms of circular RNA hsa_circ_087631 in the patients with primary biliary cholangitis （PBC）. METHODS RT-qPCR was used to detect the expression of hsa_circ_087631 in PBC patients and healthy controls. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was constructed and transfected into human acute T cell leukemia Jurkat cells， and then the expression levels of hsa_circ_087631， Bcl-6 mRNA and interleukin-21 （IL-21） mRNA were detected by RT-qPCR. Dual-luciferase reporter assay was performed to identify the interactions between hsa_circ_087631 and hsa-miR-346. RESULTS The expression of hsa_circ_087631 in the PBC patients was significantly increased compared with the healthy subjects. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was successfully constructed. The expression of hsa-miR-346 was significantly increased after the hsa-miR-346-overexpressing lentiviral vector was transfected into the Jurkat cells， while the expression levels of hsa_circ_0087631， Bcl-6 mRNA and IL-21 mRNA were significantly decreased. After wild-type or mutant hsa_circ_087631 vector and hsa-miR-346 mimics were transfected into 293T cells， the results of dual-luciferase reporter assay showed that hsa-miR-346 significantly decreased the luciferase activity of wild-type hsa_circ_087631 （P<0.01）， but the regulation did not change significantly after mutation of the predicted binding site. CONCLUSION Peripheral blood hsa_circ_087631 level is elevated in the PBC patients. The hsa_circ_087631/hsa-miR-346/Bcl-6 signaling may take effect in human T cells. Hsa-miR-346 significantly reduces the expression of hsa_circ_087631， but it may not be regulated by predicted binding sites.     

复硝酚钠与胺鲜酯对棉花化肥吸收率的影响

【目的】研究复硝酚钠(CSN)与胺鲜酯(DA-6)2种植物生长调节剂,分别和化肥复配施用对棉花化肥吸收率的影响。【方法】通过复硝酚硝与胺鲜酯分别和化肥复配施用,采集棉花2叶期、4叶期、6叶期各20株,测定棉株鲜重,干重,以及全氮、全磷和全钾含量,计算棉花3个时期的干物质比、氮吸收率、磷吸收率、钾吸收率,与CK相比较,分析复硝酚钠与胺鲜酯对棉花肥料吸收率的影响。【结果】复硝酚钠和化肥复配施肥后,棉花对肥料的吸收率在2叶期时氮、磷、钾吸收率分别提高了10.76%、0.87%和10.4%。4叶期时氮、磷、钾吸收率分别提高了18.5%、2.34%和24.86%。6叶期时氮、磷、钾吸收率分别提高了31.14%、5.33%和35.56%;胺鲜酯和化肥复配施肥后棉花对肥料的吸收率在2叶期时氮、磷、钾吸收率分别提高了8.31%、0.85%和9.25%。4叶期时氮、磷、钾吸收率分别提高了11.51%、1.48%和15.66%。6叶期时氮、磷、钾吸收率分别提高了27.11%、4.49%和30.31%。【结论】复硝酚钠与胺鲜酯与化肥复配施用,对棉花化肥吸收率有明显的促进作用。     

云南和广东省流行性出血病病毒血清6型毒株(EHDV-6)的分离与遗传特性

【目的】分离流行于我国南方地区的流行性出血病病毒血清6型毒株(EHDV-6),掌握分离毒株的遗传特征。【方法】对设立于我国云南和广东省的哨兵动物定期采血,进行EHDV感染情况监测与病毒分离培养。通过血清中和试验确定分离EHDV的血清型,采用一步法RT-PCR对分离的EHDV-6毒株基因节段2、3、6(Seg-2、-3、-6)进行扩增与序列分析。【结果】2012—2016年,从云南省和广东省的哨兵动物中分离出25株EHDV毒株,其中,11株为EHDV-6型。中国EHDV-6型毒株的Seg-2、-3与-6均属Eastern型,核酸序列相似性分别为99.1%、98.9%和98.8%,在系统发生树上分别聚为1个独立的中国支系。在日本引起疫病暴发的EHDV-6型毒株与中国EHDV-6型毒株具有最近的亲缘关系。日本EHDV-6型毒株在系统发生树上处于中国支系内部,Seg-2、Seg-3与中国毒株的核酸序列相似性分别为98.5%和93.9%。【结论】云南和广东省流行的EHDV-6型毒株核酸与氨基酸序列高度相似;日本和中国的EHDV-6型毒株具有最近的亲缘关系,提示中日之间可能存在EHDV的流动。研究结果为进一步开展我国EHDV-6型毒株流行病学分析、致病性、病原学诊断与疫苗制备等研究提供了基础。     

DTA-6对两种食用豆生理代谢及产量的影响

为探究2-N,N-二乙氨基乙基己酸酯（diethyl aminoethyl hexanoate,DTA-6）对食用豆叶片的生理代谢及产量的调控效应,选用芸豆（英国红）和小豆（龙垦2号）为试验材料,采用大田完全随机试验方法,于芸豆和小豆的初花期叶面喷施DTA-6,以喷施清水为对照（CK）,测定各生育时期叶片光合参数、碳代谢产物、干物质、产量及产量构成因素。结果表明：DTA-6提高了2种食用豆各生育期的SPAD值、盛花期和鼓粒期的净光合速率（P_n）、蒸腾速率（T_r）、气孔导度（G_s）和胞间CO₂浓度（C_i）,显著增加了叶片蔗糖和可溶性糖的含量,促进了叶片淀粉的积累;与CK相比,DTA-6提高了2种食用豆的地上部单株干物质积累量,并且显著提高了芸豆和小豆鼓粒期荚分配率;DTA-6可有效调控2种食用豆的单株荚数、单株粒数和百粒重,从而提高产量,DTA-6处理的芸豆和小豆产量分别较CK增加13.30%和12.91%,增产显著。     

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.     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

Molecular cloning and functional characterization of the fatty acid delta 6 desaturase (FAD6) gene in the sea cucumber Apostichopus japonicus

The sea cucumber (Apostichopus japonicus), an important echinodermata, had high value in nutrition and medicine for its rich collagen, sulphated polysaccharide, glycosides and polyunsaturated fatty acids (PUFA). The cDNA of the fatty acid desaturase gene in A. japonicus (AJFAD6) was cloned and was found to encode a desaturase with delta 6 FAD activity. Sequence analysis indicated that AJFAD6 included an open reading frame of 1392 bp, encoding 463 amino acids. AJFAD6 has all the conserved motifs found in other members of the FAD6 family, including an N‐terminal cytochrome b5 domain and three histidine‐rich regions. qRT‐PCR showed that AJFAD6 was expressed in all tissues tested during juvenile development and was mainly expressed in the respiratory tree at 150 days after adherence (150 days) and in the intestine at 100 days. Furthermore, AJFAD6 mRNA was also detected in the analysed adult tissues, with higher expression in the intestine and testis. Functional characterization of AJFAD6 in a recombinant yeast, Pichia pastoris, showed that AJFAD6 could catalyse exogenous linoleic acid (LA) and α‐linolenic acid (ALA) to produce γ‐linoleic acid (GLA) and stearidonic acid (STA), respectively, at conversion rates of 11.1% for LA to GLA and 3.4% for ALA to STA. Our results suggested that the biosynthetic pathway of PUFA existed in the sea cucumber, but endogenous production of eicosapentaenoic acid, arachidonic acid and docosahexaenoic acid from either LA or ALA precursor appeared to be limited.