Effects of astragaloside on the levels of sex hormone and oxidative stress injury in rats with polycystic ovary syndrome

AIM To investigate the effects of astragaloside on the levels of sex hormone and oxidative stress in rats with polycystic ovary syndrome （PCOS）. METHODS Female SD rats （n=60） were randomly divided into normal control group, model group, Diane-35 （0.339 2 mg/kg） group, low dose astragaloside （12.5 mg/kg） group and high dose astragaloside （50 mg/kg） group, with 12 rats in each group. The PCOS model was induced by letrozole （1 mg/kg）, which was administered by gavage once a day for 3 weeks. After administration, the estrus cycle of the rats was observed by vaginal smear, and the ovarian index was calculated. HE staining was used to observe the histopathological changes of the ovaries. Serum levels of the sex hormones testosterone （T）, estradiol （E₂）, luteinizing hormone （LH） and follicle-stimulating hormone （FSH） were measured by ELISA. The levels of superoxide dismutase （SOD）, malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in serum and ovarian tissue were detected by colorimetry, and the protein levels of steroidogenetic acute regulatory protein （StAR） and apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 in ovarian tissue were detected by Western blot. RESULT Compared with control group, the oestrous cycle of the rats in model group was disorder, and the ovarian index was increased, ovary was polycystic. The serum levels of T, LH and MDAwere significantly increased （P<0.05）, while the contents of E2, FSH and the activities of GSH-Px and SOD were significantly decreased （P<0.05）. The levels of MDA, StAR, cleaved caspase-3 and Bax proteins in ovarian tissue were significantly up-regulated （P<0.05）. GSH-Px and SOD activities and Bcl-2 protein levels were significantly down-regulated （P<0.05）. CONCLUSION Astragalosideeffectively balances the levels of sex hormone in PCOS rats and relieves the oxidative stress injury, the mechanism may be related to the inhibition of StAR expression.     

猕猴桃AdCCS1基因的克隆及其表达调控

采用RT-PCR法从‘米良1号’猕猴桃中分离到1 066 bp的铜锌超氧化物歧化酶分子伴侣蛋白基因AdCCS1,登录号为KY471361。AdCCS1的开放阅读框为984 bp,编码327个氨基酸,包含3个典型结构域（N端结构域、中间结构域和C端结构域）和2个保守的金属结合位点（MXCXXC和CXC）。基因结构分析显示AdCCS1由6个外显子和5个内含子组成。系统进化分析表明AdCCS1聚在双子叶植物分支中,与茶树CsCCS的亲缘关系最近。此外,AdCCS1的5′端调控区存在多种光响应、胁迫响应和激素响应应答元件。定量PCR分析显示,AdCCS1在猕猴桃叶片中的表达量最高,其次是成熟果、花、幼果和茎,在根中的表达量最低。猕猴桃果实中AdCCS1在4 ℃低温贮藏过程中的表达量均比25 ℃贮藏中同期的低,说明低温抑制AdCCS1的表达。脱落酸和赤霉素处理后AdCCS1的表达下调,但二者的应答模式不同。     

Response of CREG1 promoter to serum starvation in human vascular cells

AIM: In order to explore the regulatory mechanisms of the human cellular repressor of E1A-stimulated genes (hCREG1) in vascular smooth muscle cells (VSMCs)-HITASY and in human umbilical vein endothelial cells (HUVECs), we clone and construct hCREG1 promoter vector to detect its expression in different vascular cells. METHODS: With the results of biological information, the fragments including -3 677 bp, -2 310 bp and -945 bp upstream sequences of hCREG1 were amplified respectively using human genomic DNA template by PCR. The products were inserted into pMD18-T vector, and then were subcloned into pEGFP-1 report vector to obtain the pEGFP-hCREG1-promoter vectors. The pEGFP-hCREG1-P3677, pEGFP-hCREG1-P2310 and pEGFP-hCREG1-P945 vectors were transfected into HITASY cells and HUVECs transiently and the expression of green fluorescent protein (GFP) was detected respectively by Western blotting and fluorescent microscope. RESULTS: It was confirmed that all three PCR fragments inserted into vectors were corrected by sequencing analysis. However, the expression of GFP was different significantly in both types of vascular cells. The expression of GFP in HUVECs was higher than that in HITASY cells. Meanwhile, the expression of GFP in HITASY cells with 0.5% FBS was increased obviously compared to that in HITASY cells with 10% FBS. CONCLUSION: The reporter vectors of hCREG1 promoter are constructed successfully in which the core promoter region might be located in -945 bp-0 bp of 5′ upstream sequences. This study will provide an experimental basis for exploring the regulation of hCREG1 expression.     

苹果6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1的克隆和功能分析

以‘嘎拉’苹果为材料,克隆了6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1(序列号:MDP0000279299)全长。序列分析显示,该基因包含一个长为1 047 bp完整的开放阅读框,编码347个氨基酸,分子量为36.430 k D,预测等电点为9.24。同源性分析表明Md6PGDH1还有另外3个同源基因;功能域分析表明Md6PGDH蛋白含有两个保守的绑定域;亚细胞预测表明Md6PGDH定位存在差异。分析Md6PGDH1启动子发现存在多个响应非生物胁迫的顺式作用元件。定量分析显示,Md6PGDH1在苹果的不同组织中都有表达,且受非生物胁迫诱导。原核诱导Md6PGDH1蛋白并进行蛋白酶活的测定,为后续蛋白功能鉴定奠定了基础。Md6PGDH1在苹果愈伤组织中过量表达,提高其抗盐胁迫的能力。     

黄花菜胚囊受精前后的超微结构观察

 应用透射电镜技术研究黄花菜（Hemerocallis citrine Baroni）助细胞、卵细胞、中央细胞、 合子与初生胚乳核的超微结构特征。结果如下：助细胞有高度发达的丝状器;一个助细胞在花粉管进入 后退化;宿存助细胞在合子期退化。受精前,卵细胞、助细胞合点端无壁,细胞膜完整;受精后,合子 被完整的细胞壁包围。受精前超微结构特征显示卵细胞代谢活动低,合子期的代谢活性被激活;中央细 胞、初生胚乳核代谢活性较高。卵细胞质中的精核由双层核膜包被,染色质高度凝聚,核孔数少。助细 胞、卵细胞、中央细胞缺乏中央大液泡是黄花菜成熟胚囊的显著特征。     

花椰菜新品种雪剑一号的选育

雪剑一号是以福建地方品种经7代连续自交选育出的优良自交系CH19为父本,以雄性不育系02-9-10为母本配制成的早熟秋花椰菜一代杂种,生育期60d左右;植株生长势较强,株型直立紧凑,外叶深绿色、蜡质中等,叶柄较短,内叶合抱自护球;花球雪白、紧实、高圆形,单球质量1.12kg左右,每667m2产量2600kg左右。     

不同保鲜膜对草莓果实贮藏品质的影响

以草莓"红颜"为试材,以PVC(聚氯乙烯)膜为对照,研究了PE(聚乙烯)膜、PHM(多层PE系)膜、纳米银膜、PVDC(聚偏二氯乙烯)膜等5种保鲜处理在(4.0±0.6)℃贮藏条件下对草莓果实失重率、硬度和可溶性固形物、维生素C、可溶性糖含量及感官品质的影响。结果表明:相较于PVC保鲜膜,PVDC膜与PHM膜能够较好的保持草莓果实的贮藏品质。其中PVDC膜包装可以有效地减少果实水分散失,保持草莓的硬度以及果实可溶性固形物含量;PHM膜包装可以较好的保持草莓果实可溶性固形物、维生素C、可溶性糖含量,维持果实的营养成分。     

低温条件下溴甲烷和硫酰氟对紫穗槐豆象的毒力

熏蒸环境温度为5℃、10℃的条件下,设计6～10个不同剂量分别开展溴甲烷和硫酰氟熏蒸杀灭紫穗槐种子携带紫穗槐豆象(Acanthoscelides pallidipennis)老熟幼虫的毒力试验,应用PoloPlus软件进行机率值分析的结果表明:在5℃条件下,预计紫穗槐豆象老熟幼虫达到99.9968%死亡率所要求的CT值(Lct_(99.9968))为溴甲烷2855.7g.h/m~3、硫酰氟3973.5g.h/m~3,熏蒸24h所需剂量为119.3g/m~3、158.7g/m~3;在10℃条件下,Lct_(99.9968)为溴甲烷2145.2g.h/m~3、硫酰氟1436.3g.h/m~3,熏蒸24h所需剂量为88.1g/m~3、59.0g/m~3。因此,随着熏蒸温度的提高,硫酰氟对紫穗槐豆象老熟幼虫的毒力作用增长明显,可作为进出口紫穗槐种子检疫处理的有效熏蒸剂,但熏蒸的环境温度须高于10℃。     

香蕉枯萎病菌myosin-1基因的功能分析及外施dsRNA介导的抗性评估

 香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum f. sp. cubense,Foc)引起的香蕉毁灭性土传病害,其中 4号生理小种(Foc4)能感染几乎所有的香蕉品系,危害最严重。SMART在线软件分析myosin-1基因具有肌球蛋白马达蛋白(myosin motor domain, MMD),肌动蛋白尾结构TH1(myosin tail)和 Src家族同源结构域SH3(src homology domain 3),与禾谷镰刀菌中氰烯菌酯靶标基因myosin-5具高度的蛋白同源性,相似性高达83%。利用Split-marker基因重组技术获得Foc4的myosin-1基因敲除突变体,Δmyosin-1突变株丧失了对氰烯菌酯的敏感性,菌丝生长缓慢,产孢量减少且孢子畸形,对香蕉致病力严重下降,证实myosin-1是氰烯菌酯在Foc4中的作用靶标基因。外施靶向myosin-1体外转录的dsRNA,能抑制菌丝的生长,降低菌丝活性;菌丝膨胀扭曲分枝增多,出现典型的球状结构,与氰烯菌酯处理后的表型一致。在盆栽活体人工接种实验中,体外施用dsRNA可以明显抑制枯萎病外部症状的发展,推迟发病时间,赋予寄主抗性,结果说明体外施用dsRNA可以作为新型杀菌剂防治香蕉枯萎病。综上,myosin-1基因作为氰烯菌酯在Foc4中的靶标基因具有高度的序列保守,在调控菌丝生长发育,产孢以及致病力等方面发挥重要作用,而外施dsRNA具有防治香蕉枯萎菌的巨大潜力。     

土壤团聚体形成的研究进展

土壤团聚体是土壤结构的基本单位,它的形成与稳定是自然过程与人类活动的综合结果,即受生物与非生物因素如土地利用方式改变、耕作和种植制度等多种因素的影响。由于团聚体的形成原因及土壤环境条件不同,不同粒级的团聚体在土壤肥力上表现出不同的性质和特征。