米诺环素对糖尿病大鼠视网膜神经细胞凋亡的影响

目的：观察米诺环素对糖尿病大鼠视网膜神经细胞的凋亡的影响,研究米诺环素对糖尿病视网膜神经保护作用,对米诺环素在糖尿病视网膜疾病中抑制神经细胞凋亡提供理论支持。方法：选择健康成年雄性SD大鼠30只,随机分成正常对照组、糖尿病模型组和米诺环素治疗组,每组10只。腹腔内注射链脲佐菌素（STZ）诱发大鼠糖尿病。米诺环素治疗纽给予米诺环素腹腔注射（45mg／kg）,共注射10d,模型组和对照组腹腔注射等体积生理盐水,于给药后8周的3组动物处死,冰浴下取其视网膜组织,随后制备视网膜石蜡切片,采用末端脱氧核糖核酸介导生物素化脱氧尿嘧啶缺口末端标记L（TUNEL）法进行凋亡细胞原位标记,行视网膜神经细胞凋亡计数,对所有数据进行统计学分析。实验结果拟用均数和标准差（x±s）表示,P〈0．05为差异有统计学意义。结果 相同观察时相,与阴性对照组比较,模型对照组大鼠视网膜TUNEL阳性细胞显著增多（P〈0．01）,在米诺环素组,大鼠视网膜TUNEL阳性细胞数比模型对照组明显减少,差异有统计学意义（P〈0．01）。结论：米诺环素能有效降低糖尿病大鼠视网膜神经细胞的凋亡,对糖尿病视网膜神经细胞有保护作用。     

青蒿琥酯对糖尿病视网膜病变组织中Bcl-2和Hsp27表达的影响

为了探讨青蒿琥酯(artesunate,ART)对糖尿病视网膜病变组织中抗凋亡和细胞保护作用相关因子Bcl-2和热休克蛋白27(Hsp27)表达的影响,将30只健康雄性SD大鼠随机分为空白对照组(negative control)、模型对照组(model control)及药物治疗组(ART treated),其中模型对照组和药物治疗组给予注射链脲佐菌素(STZ)建立1型糖尿病大鼠模型,饲养3个月后,药物治疗组大鼠腹腔注射青蒿琥酯治疗10 d,分离三组大鼠视网膜,提取蛋白和总RNA。通过Western-blot,实时荧光定量PCR(RT-q PCR)对Bcl-2和Hsp27的表达情况进行检测。结果显示:Bcl-2在模型对照组中大鼠视网膜的表达下降(p0.05),药物治疗组则显著增加(p0.01);Hsp27在模型对照的表达量高于空白对照组(p0.05),治疗组的表达量显著高于模型对照组(p0.01)。结论经青蒿琥酯治疗后糖尿病大鼠视网膜组织中抗凋亡因子Bcl-2和细胞保护因子Hsp27表达均增强,减少糖尿病视网膜细胞凋亡,对神经细胞具有保护作用。     

枸杞多糖对糖尿病大鼠视网膜神经元的保护作用及其机制

目的:观察枸杞多糖(LBP)对糖尿病大鼠视网膜神经细胞的保护作用,并探讨其作用机制。方法:18只SD大鼠随机分为3组(n=6):正常对照组(NC),糖尿病模型组(DM)和LBP治疗组(DM+LBP),通过一次性腹腔注射链脲佐菌素(STZ)的方法制备糖尿病大鼠模型。DM+LBP组按1 mg/(kg·d)剂量的LBP灌胃12周。治疗结束后检测大鼠体重、空腹血糖、视网膜活性氧簇(ROS)的生成、视网膜神经节细胞(RGCs)和无长突细胞的表达、视网膜NF-E2相关因子2(Nrf2)和血红素加氧酶-1(HO-1)的蛋白表达。结果:STZ诱导糖尿病大鼠模型造模成功率100%。与NC组相比,DM组大鼠体重明显降低、空腹血糖值升高、ROS的生成明显增加、RGCs和无长突细胞的数量均明显减少(P<0.01)。与DM组相比,LBP治疗组大鼠体重升高、血糖降低、ROS的生成减少、RGCs和无长突细胞的数量均明显增加(P<0.01或P<0.05);视网膜Nrf2和HO-1的蛋白表达均明显升高(P<0.01)。结论:LBP能改善糖尿病大鼠视网膜的氧化应激状态,对糖尿病大鼠视网膜神经细胞有一定的保护效应,其作用机制可能与其激活Nrf2/HO-1信号通路有关。     

大鼠脑创伤后caspase-3的过度表达与细胞凋亡的关系

目的:探讨大鼠脑创伤后海马神经组织中casepase-3表达及其在细胞凋亡中的机制。方法:雄性Wistar大鼠72只随机分成对照组和创伤组,用Marmarou方法造成大鼠重型弥漫性颅脑创伤,采用免疫组织化学检测海马CA1区神经细胞casepase-3蛋白表达情况,原位细胞DNA断裂检测末端标记(TUNEL)法观察大鼠海马CA1区神经细胞凋亡动态变化。同时行TUNEL与caspase-3双标染色。结果:对照组海马区神经细胞casepase-3未见明显表达,创伤组海马CA1区神经细胞casepase-3表达在伤后3小时开始升高,伤后3天达高峰(P0.01),伤后7天下降明显。对照组海马区未见TUNEL阳性细胞,创伤组海马区TUNEL阳性细胞伤后3小时开始增多,伤后3天达高峰(P0.01),伤后7天下降。可见创伤组TUNEL染色与caspase-3免疫染色双标阳性的细胞伤后6小时细胞数量逐渐增多,于伤后3天达高峰(P0.01),伤后7天双标阳性细胞数量下降。Casepase-3表达与TUNEL阳性细胞明显相关(P0.01)。结论:大鼠脑创伤后casepase-3的过度表达是影响大鼠脑创伤后神经细胞凋亡原因之一,抑制casepase-3活性表达对神经组织起保护作用。     

七叶皂苷钠对一氧化碳中毒迟发性脑病大鼠模型细胞凋亡 及调控凋亡基因表达的影响

目的:探讨七叶皂苷钠对一氧化碳中毒迟发性脑病大鼠模型细胞凋亡及Caspase-3表达的影响。方法:将30只大鼠随机分为正常对照组、一氧化碳中毒迟发性脑病大鼠模型对照组和七叶皂苷钠治疗组,原位末端标记法(TUNEL)检测细胞凋亡,免疫组化法检测Caspase-3的表达。结果:治疗组大鼠脑细胞凋亡率及Caspase-3表达较迟发性脑病对照组显著降低(P<0.01)。结论:七叶皂苷钠对一氧化碳中毒迟发性脑病的脑保护作用机制可能与干预脑细胞凋亡相关基因表达并减少神经细胞凋亡有关。     

米诺环素对不完全脊髓损伤大鼠脊髓BDNF和NT-3表达的影响

目的观察腹腔注射米诺环素对改良Allen’s法造成的不完全脊髓损伤大鼠脊髓中脑源性神经营养因子以及神经营养因子3表达的影响,探讨米诺环素治疗脊髓损伤的作用机制。方法成年雌性Sprague-Dawley(SD)大鼠54只,改良Allen’s法造成不完全脊髓损伤,根据实验需要可以分为3组,空白组,只打开脊柱椎板,不损伤;治疗组,大鼠脊髓损伤,并腹腔注射米诺环素;损伤组,大鼠脊髓损伤,腹腔注射等剂量的生理盐水。观察各组大鼠的后肢能力Basso-Beattie-Bresnahan评分,并于不同时段(3d、7d,14d)取大鼠脊髓T8-9段采用逆转录PCR,以及免疫化学组织染色法测定脑源性神经营养因子以及神经营养因子3的表达。结果米诺环素能够明显改善不完全脊髓损伤大鼠的功能,逆转录PCR和脊髓组织冰冻切片免疫组织化学染色DAB都能证实米诺环素治疗组脑源性神经营养因子以及神经营养因子3表达显著增多。结论米诺环素在治疗不完全脊髓损伤大鼠的机制还应与其上调了大鼠体内的脑源性神经营养因子以及神经营养因子3表达有关。     

透明质酸联合盐酸米诺环素对大鼠金黄色葡萄球菌感染创口的治疗效果

目的:探讨透明质酸联合盐酸米诺环素对大鼠金黄色葡萄球菌感染创口的治疗效果。方法:选择32只Wistar大鼠,在背部脊柱两侧各制备1个全层皮肤缺损模型。创面接种细菌形成感染创口并将大鼠随机地分为4个组,每个组共有16个创口。实验组(1.5%透明质酸联合0.1%盐酸米诺环素凝胶治疗组)、对照组1(0.9%生理盐水治疗组)、对照组2(1.5%透明质酸凝胶治疗组)、对照组3(0.1%盐酸米诺环素凝胶治疗组)。观察分析创口愈合情况及创口愈合率,HE染色组织形态学分析,免疫组织化学染色法分析创口中肿瘤坏死因子α的表达。结果:透明质酸联合盐酸米诺环素组创口愈合率为(78.13±3.04)%,显著高于生理盐水对照组、透明质酸对照组和盐酸米诺环素对照组(均P0.05);透明质酸联合盐酸米诺环素组创口可见大量成熟的纤维组织,生理盐水组创口可以观察到组织发生坏死,结构不清;透明质酸组可见富含血管的新生肉芽组织;盐酸米诺环素组可见大量纤维组织;透明质酸联合盐酸米诺环素组创口TNF-α表达为10.84±1.49,显著低于生理盐水对照组、透明质酸对照组和盐酸米诺环素对照组(均P0.05)。结论:1.5%透明质酸联合0.1%盐酸米诺环素凝胶对大鼠金黄色葡萄球菌感染创口有较好的治疗效果。     

西酞普兰对慢性应激大鼠额叶皮质神经细胞bax mRNA、bcl-2 mRNA表达及凋亡的影响

目的:探讨西酞普兰对慢性应激大鼠的额叶皮质神经细胞bax、bcl-2 mRNA表达的影响。方法:取24只雄性SD大鼠随机分为对照组(不进行任何处理)、应激组(应激+生理盐水灌胃)、实验组(应激+西酞普兰灌胃),采用强迫游泳制造慢性应激模型(15 min/d,共4周),用原位杂交技术检测bax、bcl-2 mRNA表达情况,TUNEL法检测细胞凋亡,尼康图像分析(NIS DR)软件测量各指标阳性细胞数量。结果:应激组较对照组,大鼠额叶皮质bax mRNA阳性表达细胞明显增多(P0.01)、染色加深;bcl-2 mRNA阳性表达细胞明显减少(P0.01)且染色变浅,且TUNEL阳性细胞数量增多(P0.01),染色增强。而实验组较应激组大鼠,额叶皮质bax mRNA阳性表达细胞减少(P0.01)、染色变浅,bcl-2 mRNA阳性表达细胞明显增多(P0.05),染色加深,且TUNEL阳性细胞数量减少(P0.01),染色减弱。结论:大鼠额叶皮质神经细胞bax mRNA和bcl-2 mRNA的表达水平受到慢性应激的影响,导致细胞凋亡加剧,西酞普兰能够有效调控额叶皮质神经细胞bax和bcl-2 mRNA的表达水平,拮抗细胞凋亡,这可能是西酞普兰防治慢性应激引起的神经精神疾病的相关机制之一。     

雷公藤甲素对病毒性心肌炎细胞凋亡及Fas/FasL 蛋白表达的研究

目的:研究雷公藤甲素对柯萨奇病毒B3病毒(CVB3)感染的病毒性心肌炎小鼠心肌细胞凋亡和Fas/FasL蛋白表达的抑制作用,探讨TP治疗病毒性心肌炎的作用机制。方法:将Balb/c小鼠随机分成4组作为动物模型,分别为对照组、模型组、利巴韦林组和TP组。对照组腹腔注射生理盐水,其余三组腹腔注射CVB3,利巴韦林组和TP组小鼠分别予以相应的药物治疗后,测定各组小鼠存活率及心肌病变积分,采用末端转移酶标记技术(TUNEL法)检测小鼠心肌细胞凋亡,免疫组化法检测Fas/FasL蛋白阳性表达。结果:空白对照组心肌无病变,利巴韦林组、TP组与模型组相比有显著性差异(P<0.01)。正常组鲜见心肌细胞凋亡,模型组细胞凋亡率较正常组显著增加(P<0.01),治疗组利巴韦林组和TP组凋亡率比模型组明显降低(P<0.05,P<0.01)。模型组Fas/FasL表达比正常组显著增多(P<0.01),治疗组利巴韦林组和TP组较模型组显著降低(P<0.01)。结论:雷公藤甲素具有通过抑制Fas/FasL蛋白的表达,减缓心肌细胞凋亡,达到抑制病毒性心肌炎从而保护心肌细胞的作用。     

VEGF165b对糖尿病大鼠视网膜神经节细胞保护作用的研究

目的:VEGF165b是新发现的血管内皮生长因子的变构体之一,本研究将观察其对糖尿病大鼠视网膜神经节细胞的抗凋亡作用.方法:采用四氧嘧啶诱发糖尿病大鼠模型,分为正常对照组(CON),糖尿病组(DM),糖尿病VEGF165b低剂量治疗组(DMT1)、中剂量治疗组(DMT2),糖尿病高剂量治疗组(DMT3),糖尿病单纯胰岛素治疗组(DMT4),所有治疗组在糖尿病成模后1个月开始治疗.2个月后处死各组大鼠,摘取眼球进行光镜形态学观察、核苷酸末端转移酶介导的dUTP缺口翻译法(TUNEL法)视网膜神经节细胞凋亡检测.结果:VEGF165b治疗使糖尿病大鼠视网膜光镜形态学改变减轻,能有效的抑制视网膜神经节细胞凋亡.VEGF165b治疗组视网膜神经节凋亡细胞数较DM组明显减少(P＜0.01),与糖尿病大鼠单纯胰岛素治疗组相比差异也有统计学意义.随着VEGF165b浓度的增加视网膜神经节细胞凋亡个数减少,但1ng/μL组与10ng/μL组相比差异无统计学意义.结论:VEGF165b对视网膜神经节细胞有保护作用,可能对糖尿病视网膜病变具有治疗有意义.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The mechanism of hatching of eggs of Haemonchus contortus

Fluid collected from hatching eggs of Haemonchus contortus contained a lipase which hydrolysed 2-naphthyl laurate (about 0·7 μmol naphthol freed /h/10⁶ eggs). The fluid also hydrolysed l-leucinamide (about 2·3 μmol leucine freed/h/10⁶ eggs). The fluid when added to normal or heated eggs caused ‘hatching’. ‘Hatching’ also occurred in exsheathing fluid from infective juveniles and in a preparation of pancreatic lipase containing leucine aminopeptidase. A purified mammalian leucine aminopeptidase in combination with several different lipases did not attack egg shells.The ‘spontaneous’ hatching of eggs of H. contortus was strongly inhibited by 1,10-phenanthroline, 10^?3M, and this inhibition was reversed by Zn²⁺. However, the inhibition of ‘hatching’ of eggs in externally applied hatching fluid, or the hydrolysis of leucinamide in hatching fluid was generally less marked.