原矛头蝮蛇毒对小鼠器官形态的影响

目的:观察原矛头蝮(P.mucrosquamatus)蛇毒对小鼠脏器形态结构的影响,从组织学角度探讨该蛇毒的致病机理。方法:用原矛头蝮攻击小鼠,待小鼠死亡后,立即进行解剖,观察心脏和大血管内部血栓形成情况。取心脏、血管、周围神经、肾脏、肝脏、脾脏和肺,观察外形,并进行石蜡切片,H.E染色观察。用健康小鼠相同器官为对照,比较二者的差异。结果:被原矛头蝮攻击致死小鼠的心脏、肝脏和肺发生了显著收缩;心腔和血管腔被血栓堵塞;在心脏、肝脏和肾脏组织血管中观察到了血栓;脾脏中白细胞显著增多;肺泡收缩,细支气管严重收缩致管腔堵塞;股动脉血管内膜发生了脱落;在股神经中发生了脱髓鞘现象。结论:急性凝血和窒息是导致小鼠死亡的重要原因;原矛头蝮蛇毒兼具血循毒和神经毒的特征;原矛头蝮蛇毒可导致多种脏器内部结构发生改变。     

广西发现茂兰原矛头蝮及分类探讨

2011年10月,相继在广西弄岗国家级自然保护区采得2号雄性茂兰原矛头蝮Protobothrops maolanensis,为广西自治区爬行类新纪录。这2号标本基本符合茂兰原矛头蝮鉴别特征,但有些特征又与越北原矛头蝮P. trungkhanhensis相似,提示两个物种的分类关系还需进行深入研究。     

浙江省发现角原矛头蝮

<正>2014年7月24日,于浙江省台州市仙居县(28°37′N,120°36′E,海拔339 m)发现1号雄性活体蛇类标本,经查阅文献和鉴定,确认为角原矛头蝮(Protobothrops cornutus),系华东地区及浙江省蛇类属、种的地理新分布,也是我国发现角原矛头蝮仅有的第5个分布点。本次采集的角原矛头蝮标本全长615.0 mm,头长26.0 mm,体长484.0 mm,尾长105.0 mm。头呈三角形,头颈部区分明显,上颌1对管牙,鼻眼间有颊窝,颊窝由3片大鳞围成,其中1枚为第2上唇鳞。上唇鳞10枚,鼻鳞与第1上唇鳞分离。下唇鳞14枚。舌前段2/3为黑色,后     

河南省驻马店发现大别山原矛头蝮

<正>2016年9月17日,河南省驻马店市西平县村民王孝在自家果园中(33°11′32.45″N,113°37′19.42″E,海拔218 m)采集到1号蛇类标本,置于75%酒精保存后,送至沈阳师范大学两栖爬行动物研究所,经鉴定为蝰科(Viperidae)蝮亚科(Crotalinae)原矛头蝮属(Protobothrops)大别山原矛头蝮(P.dabieshanensis),为河南省蛇类新纪录种,     

广东天井山林区发现角原矛头蝮

2009年4～7月,在广东天井山林区采集到3条角原矛头蝮(Protobothrops cornutus),这是该蛇在我国野外第二次被发现,天井山是新发现的一个分布点。本次野外新发现进一步证实了角原矛头蝮在我国的自然分布,为深入了解该蛇的形态特征及地理分布格局提供了重要基础资料。     

雀嘴茶半数致死量及病理学的初步研究

观察了云南广泛使用的雀嘴茶对小鼠的急性毒性反应及病理变化,为雀嘴茶使用安全性提供可靠的科学依据,我们以一定浓度不同体积的提取物给小鼠灌胃给药,连续观察小鼠行为活动、毒性反应程度、及死亡情况等指标,并及时对小白鼠进行病理切片观察,以获得初步毒性、病理资料。结果显示雀嘴茶半数致死量LD50为39．9g／kg,说明市售用于保健作用的雀嘴茶有一定的毒性,病理切片也说明其对胃肠道、心、肝、脑等多个脏器都有一定的病理损伤,长期使用可能会对人体造成一定的危害,应该慎重使用,甚至是禁止使用。     

东亚钳蝎粗毒蛋白质热稳定性的研究

目的:通过蝎毒热溶解性和毒性实验,探索蝎毒蛋白稳定性与分子量间的关系。方法:对蝎毒粗毒煮沸后的可溶成分进行急性毒力实验和SDS-PAGE电泳分析。结果:蝎毒粗毒加热处理组的LD50为16.228mg/kg,粗毒对照组的LD50为3.66mg/kg,显示蝎毒经加热后毒力明显降低,但仍具有一定的毒性。SDS-PAGE结果显示,经加热处理的粗蝎毒之沉淀主要为大分子蛋白质,在可溶成分中含有小分子肽类。结论:蝎毒的大分子蛋白质热稳定性差,而小分子多肽部分耐热性强,而且蝎毒的小分子多肽组分具有相对稳定的毒性。     

红尾蚺和原矛头蝮基因组微卫星分布特征比较分析

本研究分析比较了红尾蚺Boa constrictor和原矛头蝮Protobothrops mucrosquamatus基因组微卫星的分布特征,通过MISA分别鉴定出398 860个和422 364个微卫星,其长度分别为8 550 741 bp和12 243 226 bp,分别占基因组序列总长度的0.59%和0.73%,在各自基因组中的丰度分别为275.46个/Mbp和252.33个/Mbp。红尾蚺基因组中单碱基重复类型微卫星最多,其次是四碱基、二碱基、三碱基、五碱基和六碱基,最丰富的5种微卫星类型是A、AC、AAAT、AG、AAT;原矛头蝮基因组中单碱基重复类型微卫星最多,其次是三碱基、四碱基、二碱基、五碱基和六碱基,最丰富的5种微卫星类型是A、AAT、AC、C、AAAT。红尾蚺和原矛头蝮微卫星在基因组不同区域丰度不同,基因间区丰度最高,其次是内含子和外显子,编码区微卫星丰度最低,表明编码区微卫星受到的选择压力最大。红尾蚺和原矛头蝮在基因中微卫星丰度分布的位置特征相似,即微卫星在基因上下游500 bp丰度最高,在内含子次之,在外显子最低。红尾蚺和原矛头蝮基因编码区所有6种重复类型微卫星中,三碱基重复类型占绝对优势。红尾蚺和原矛头蝮基因组中含有微卫星的编码序列分别有1 480条和1 397条,被GO注释的分别有736条和733条。它们的GO功能归类结果类似,但是与其他物种相比存在种系差异。本研究结果为后续开发这2种蛇的高质量微卫星标记提供了方便,也为进一步探索这些微卫星在它们基因组中的生物学功能提供了有意义的基础数据。     

中国一新纪录种--缅北原矛头蝮(爬行纲,蛇亚目,蝰科)

记述了采自西藏Protobothrops属标本1条,该蛇原报道于缅甸北部,所获标本为中国新纪录种.标本保存在沈阳师范大学化学与生命科学学院标本馆.     

牛磺胆酸钠在大鼠急性胰腺炎模型中的半数致死量(LD50)研究

目的:通过使用不同浓度的牛磺胆酸钠(TC)建立急性胰腺炎(AP)模型,观察相应的死亡率,进而计算其半数致死量。方法:使用不同浓度的TC(0%(wt/vl)、1%、2%、3%、4%、5%、9%)观察AP造模72小时后各组的死亡率。同时分析TC浓度与组织病理评分、血清淀粉酶和外周血细胞变化的相关性。结果:TC的半数致死量为3.409%。并且,TC的浓度与组织病理评分、血清淀粉酶和外周血细胞变化密切相关。结论:TC所致AP造模的最佳剂量为3.409%。     

湖南烙铁头蛇毒蛋白组分的双向电泳分析

烙铁头蛇是世界上剧毒的蛇种之一,其所携带的毒素能够导致严重的机体损伤。应用蛋白质双向电泳技术,对湖南烙铁头蛇蛇毒蛋白的蛋白质组分进行分析。通过等电聚焦和SDS-PAGE凝胶电泳分析获得完整的烙铁头蛇毒全蛋白质的图谱,经胶体考马斯亮蓝染色后,应用PDQuest软件对蛋白表达谱进行分析。通过等电聚焦和SDS-PAGE凝胶电泳有83个蛋白质组分被检测出来。其中大约90.00%的蛋白质的相对分子质量(Mr)分布在15～45 kDa之间,大约72.29%的蛋白质等电点(pI)在4.0～7.0之间。通过对烙铁头蛇毒的蛋白组学研究,获得其蛇毒蛋白质组分的表征特点,为后续进一步研究各组分的身份和潜在功能奠定基础,既可以提出新的治疗方案又可以为新的药理应用提供宝贵资源。     

PLGA-奥沙利铂缓释微球急性毒性实验

目的了解PLGA-奥沙利铂缓释微球和奥沙利铂小鼠皮下给药的急性毒性。方法采用Bliss法分别测定PLGA-奥沙利铂缓释微球和奥沙利铂小鼠皮下给药对动物的半数致死量（LD50）。结果PLGA-奥沙利铂缓释微球小鼠皮下给药时的LD50为660.90 mg/kg,奥沙利铂小鼠皮下给药时的LD50为15.24 mg/kg,相差达43.37倍。结论PLGA-奥沙利铂缓释微球能够显著降低药物对动物的急性毒性。     

牛磺胆酸钠在大鼠急性胰腺炎模型中的半数致死量（LD50）研究

目的：通过使用不同浓度的牛磺胆酸钠（Tc）建立急性胰腺炎（AP）模型,观察相应的死亡率,进而计算其半数致死量。方法：使用不同浓度的TC（0％（wt／v1）、1％、2％、3％、4％、5％、9％）观察AP造模72小时后各组的死亡率。同时分析TC浓度与组织病理评分、血清淀粉酶和外周血细胞变化的相美性。结果：TC的卓数致死量为3．409％。并且,TC的浓度与组织病理评分、血清淀粉酶和外周血细胞变化密切相关。结论：TC所致AP造模的最佳剂量为3．409％。     

Genetic diversity and sex‐biased dispersal in the brown spotted pitviper (Protobothrops mucrosquamatus): Evidence from microsatellite markers

Dispersal plays a vital role in the geographical distribution, population genetic structure, quantity dynamics, and evolution of a species. Sex‐biased dispersal is common among vertebrates and many studies have documented a tendency toward male‐biased dispersal in mammals and female‐biased dispersal in birds. However, dispersal patterns in reptiles remain poorly understood. In this study, we explored the genetic diversity and dispersal patterns of the widely distributed Asian pitviper Protobothrops mucrosquamatus. In total, 16 polymorphic microsatellite loci were screened in 150 snakes (48 males, 44 females, 58 samples without sex information) covering most of their distribution. Microsatellite analysis revealed high genetic diversity in P. mucrosquamatus. Bayesian clustering of population assignment identified two major clusters for all populations, somewhat inconsistent with the mitochondrial DNA phylogeny of P. mucrosquamatus reported in previous research. Analyses based on 92 sex‐determined and 37 samples of P. mucrosquamatus from three small sites in Sichuan, China (Mingshan, Yibin, and Zizhong) consistently suggested female‐biased dispersal in P. mucrosquamatus, which is the first example of this pattern in snakes. The female‐biased dispersal patterns in P. mucrosquamatus may be explained by local resource competition.     

30种植物杀鼠活性研究

采用饲料混毒法测定了30种植物的杀鼠活性,用灌胃法测定了4种植物的毒性,并对2种植物中的活性成分进行了分离.结果表明苦参、曼陀罗、牛皮消、牛心朴、铁棒锤、皂荚、黄花烟草、接骨木、乌头、大戟、宽裂乌头、野八角、短柄乌头、贯众、瑞香狼毒在饵料中添加量为20%时,试鼠死亡率均在50%以上.曼陀罗、皂荚、乌头、大戟对试鼠的致死中量(LD50)分别为7.04、2.21(皂荚乙醇提取液)和3.52(皂荚丙酮提取液)、10.67和19.56 g·kg-1.同时从苦参、曼陀罗中分离出了8种单体化合物,其中7种具有较好的杀鼠活性,5种杀鼠效果显著.     

Genetic variation of the VP1 gene of the virulent duck hepatitis A virus type 1 (DHAV-1) isolates in Shandong province of China

To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1 (DHAV-1) isolates, the virulence, cross neutralization assays and the complete sequence of the virion protein 1 (VP1) gene of nine virulent DHAV-1 strains, which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007–2008, were tested. The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses (ELD₅₀s) and the median lethal doses (LD₅₀s), respectively. The results showed that the ELD₅₀s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 10⁶/mL to 1.44 × 10⁷/mL, while the LD₅₀s were 2.39 × 10⁵/mL to 6.15 × 10⁶/mL. Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates, respectively. Compared with other virulent, moderate virulent, attenuated vaccine and mild strains, the VP1 genes of the 9 strains shared 89.8%–99.7% similarity at the nucleotide level and 92.4%–99.6% at amino acid level with other DHAV-1 strains. There were three hypervariable regions at the C-terminus (aa 158–160, 180–193 and 205–219) and other variable points in VP1 protein, but which didn’t cause virulence of DHAV-1 change.     

Toxicity,Safety, and Efficacy Studies on Mesenchymal Stem Cells Derived from Decidua basalis in Wistar Albino Rats by Intravenous and Subcutaneous Routes

Ex vivo expanded decidua-basalis(DB)-derived mesenchymal stem cells (MSCs) obtained from single donors have demonstrated therapeutic benefits in in vitro and in vivo studies. In this report, the intravenous and subcutaneous administration of DB-MSCs obtained from five healthy donors was assessed considering clinical grade proliferation, accessibility, and toxic effects in Wistar albino rats. The ability of the obtained DB-MSCs for differentiating, as well as their expression of several cell surface markers and immunomodulatory activities, were all assessed. Clinical standard proliferated cells were administered to animals intravenously and subcutaneously in a series of preclinical models in order to assess their in vivo toxicity, general safety, and tumorigenic possibilities. We established that DB cells exhibit structural and functional traits with MSCs. At various doses supplied intravenously or subcutaneously, the research showed no fatality, abnormal response to therapy, or substantial pathological modifications in the rats. Furthermore, there was no indication of prenatal damage in the same animal species when the rats were repeatedly treated with DBMSCs. Thus, DBMSCs were demonstrated to be non-toxic, non-teratogenic, and non-tumorigenic. To determine whether they can be administrated to human patients without risk, more investigation is recommended.     

The variability of Vipera ammodytes ammodytes venoms from Croatia--biochemical properties and biological activity

Vipera ammodytes ammodytes venom has been used for many years in Croatia for immunization of horses and production of specific therapeutic anti-venoms. The neutralizing effectiveness of anti-venoms is directly dependent on the properties of the snake venom used for immunization. Therefore, appropriate characterization of the whole venom is necessary prior to use in the immunization procedure. In the course of such analyses, the variability in biochemical properties and biological activity was observed in venoms collected from snakes originating from different parts of Croatia. The venom pools also differed with respect to time of snake collection (1992-2003). Analyses of three samples of whole venom pools were carried out revealing differences in lethal activity (LD50), minimum haemorrhagic dose (MHD), minimum necrotizing dose (MND), phospholipase A2 activity and in anticomplementary activity. SDS-PAGE electrophoretic patterns were similar, but not identical, for all tested venom pools with respect to the number of protein bands detected, but intensity of particular components differed. Preliminary immunogenicity testing in terms of determination of specific antibodies revealed similar immunogenicity and high cross-reactivity for three samples tested.