马兜铃水提液对斑马鱼胚胎的致畸作用和心脏毒性的研究

目的：探索马兜铃水提液对斑马鱼胚胎的致畸作用和心脏毒性.方法：分别用不同浓度的马兜铃水提液和马兜铃酸A（AA）处理斑马鱼胚胎,观察致畸作用和对心脏发育影响.结果：给药组的斑马鱼胚胎出现畸形和死亡;当水提液中AA含量为0.5 μg/mL时,胚胎心率明显减慢;AA含量为5μg/mL时,胚胎在24～48 hpf之间全部死亡;水提液的LC50为1.43 μg/mL.结论：与AA相比,马兜铃水提液对斑马鱼胚胎有着更强的致畸和心脏毒性,且毒性作用具有时间和浓度依赖性.     

迷迭香酸对硫酸铜致斑马鱼胚胎毒性的抑制作用

本文探讨硫酸铜(CuSO_4)对斑马鱼(Danio rerio)胚胎发育的毒性效应,使用迷迭香酸(RA)抑制CuSO_4对斑马鱼胚胎发育的毒性并探讨其作用机制。收集受精后1 h(1 hpf)的斑马鱼胚胎暴露于不同浓度的CuSO_4溶液,或含有不同浓度迷迭香酸的CuSO_4溶液,对照组培养在E3培养液中,观察胚胎死亡、孵化及畸形情况,计算胚胎死亡率、孵化率和畸形率;以活性氧(ROS)荧光探针DCFH-DA染色法检测迷迭香酸保护下胚胎的活性氧水平。对实验数据进行方差分析。结果显示:(1)CuSO_4浓度超过一定量时能诱导斑马鱼胚胎死亡和畸形,胚胎孵化率也降低。CuSO_4对96 hpf斑马鱼胚胎的半致死浓度(LC50)为7.7μmol/L,半致畸浓度(EC50)为1.9μmol/L。(2)在96 hpf,迷迭香酸与8μmol/L CuSO_4共同处理组斑马鱼胚胎的死亡率明显降低,孵化率升高。迷迭香酸与1.6μmol/LCuSO_4共同处理组斑马鱼胚胎的畸形率降低。(3)CuSO_4单独处理组的活性氧含量明显高于迷迭香酸与CuSO_4共同处理组和对照组。结果表明,CuSO_4暴露对斑马鱼胚胎发育的毒性效应可能与活性氧升高导致的氧化应激相关;迷迭香酸抑制CuSO_4对斑马鱼胚胎发育的毒性作用,可能与减少活性氧生成有关。     

铅和邻苯二甲酸二丁酯联合暴露对斑马鱼胚胎的影响

为了研究铅(Pb)和邻苯二甲酸二丁酯(DBP)单独及联合暴露对斑马鱼胚胎神经发育和细胞凋亡相关基因表达的影响,考察两者对斑马鱼胚胎神经系统的毒性作用。将720个斑马鱼胚胎按3×3析因设计随机分为9组,以Pb(0、0.01 mg/L和1 mg/L)和DBP(0、0.005 mg/L和0.5 mg/L)单独及联合对斑马鱼胚胎暴露120 h。观察、记录胚胎的孵化、死亡情况,采用荧光定量PCR测定斑马鱼胚胎神经发育相关基因(NR1A、NR2A、NR2D)和神经细胞凋亡相关基因(bcl-2、c-fos)的表达变化情况。与空白对照组相比,各暴露组均可致斑马鱼胚胎孵化率降低、死亡率增高(P0.05);联合暴露组NR1A、NR2A、NR2D、c-fos基因表达水平显著升高,bcl-2基因表达水平显著下降(P0.05)。Pb和DBP均可影响斑马鱼胚胎的发育,且具有一定的神经毒性,而二者联合暴露毒性复杂,对斑马鱼胚胎孵化率和死亡率的影响有交互作用,对神经发育和细胞凋亡相关基因无交互作用,其机制有待进一步研究。     

氯丙嗪在斑马鱼胚胎和早期幼鱼发育过程中的神经毒性作用

目的 采用模式动物斑马鱼作为研究对象,观察氯丙嗪（chlorpromazine,CPZ）暴露对胚胎和幼鱼早期神经发育的影响.方法 在一般毒性评价的基础上,通过整体胚胎细胞凋亡检测和脑组织病理学检查,了解CPZ对神经发育的器质性改变;采用神经行为学方法,包括幼鱼触动逃避反应、自发运动以及惊恐逃避反射等,研究氯丙嗪暴露所致的神经发育功能性障碍.结果斑马鱼胚胎受精后6 h（6 hpf）～72 hpf暴露于CPZ（≥5 mg/L）可引起胚胎和幼鱼死亡、致畸和幼鱼孵化延迟,并呈浓度和时间依赖性;采用吖啶橙染色检测36 hpf整体胚胎凋亡细胞,发现凋亡细胞主要集中在胚胎中脑、后脑、丘脑以及中后脑连接区、脊索和尾部等处;脑组织病理学检测发现,7dpf幼鱼颅腔增大、脑体积减小、脑细胞缩小且细胞间隙增宽.6～72 hpf CPZ（≥0.0625 mg/L）暴露后,幼鱼神经行为学研究发现,CPZ（≥0.125 mg/L）可引起3dpf幼鱼触觉运动能力下降;CPZ（≥0 5 mg/L）可浓度依赖性地抑制幼鱼自发运动,并出现僵直不动、震颤或快速刻板式转圈运动等行为改变;光惊恐实验中,暗环境下各暴露组幼鱼对突发强光刺激均表现为惊跳逃避,并且暗-光交替期运动加速度变化与对照组无显著差异;在撤除光源后,1mg/L和2 mg/L暴露组幼鱼暗适应时程缩短,而0.125 mg/L和0.25 mg/L暴露组暗适应时程延长,提示CPZ对外界刺激引发的幼鱼活跃游动有抑制和促进双重毒性作用.结论 CPZ暴露对斑马鱼胚胎和幼鱼具有明显的神经发育毒性作用.模式动物斑马鱼作为一种高通量筛选模型在外源性化合物神经发育毒性评价中具有较好的应用前景.     

斑马鱼胚胎发育技术在毒性评价中的应用

对斑马鱼（Brachydanio rerio)胚胎发育技术在环境科学领域的应用作一综述,斑马鱼胚胎毒性技术是各国际标准组织认可的标准毒性测定方法之一,属于致畸效应检验,该项技术成本低、易操作、灵敏度高,特别是具有可记录多项毒性指标的特点,并可以此判断污染物的致毒机理,斑马鱼胚胎发育过程受重金属影响较大,其中Cu的毒性最强,Hg次之,Cr最弱,有机农药中三苯基锡类（TPTA）的毒性最强,林丹次之;有机试剂中含卤素取代基和苯胺类毒性最大,这与其它毒性测定方法的结果完全一致,并表现出较高的灵敏度,特别是选用非致死性的指标,可以初步认定这对测定复合污染物毒性并分析毒物的致畸效应方面有很好的发展前景。     

四种新型农药对斑马鱼胚胎发育的毒性效应

为了研究4种农药(腈苯唑、吡唑醚菌酯、恶霜灵、戊菌隆)单独及联合作用对斑马鱼胚胎发育的影响,设计7个实验组(空白组、溶剂组、4种农药单独暴露组及联合暴露组)对斑马鱼胚胎暴露72 h,结果显示,浓度50 ng/L的4种农药单独暴露导致心包囊和卵黄囊水肿比率、脊柱弯曲率均显著高于对照组;腈苯唑、恶霜灵及联合暴露导致斑马鱼仔鱼心率显著下降,出现显著的心律不齐;斑马鱼胚胎的Na~+/K~+-ATP酶和Ca~(2+)-ATP酶活性被显著诱导,而联合暴露表现出显著的抑制作用。结果说明,环境浓度的农药暴露对鱼类胚胎发育具有明显的毒性。     

顺铂和噪声联合作用对听觉器官的影响

研究了顺铂与噪声联合作用对耳蜗的毒性,发现联合作用后听阈偏移大于单独顺铂作用后阈移和单独噪声作用后阈移的总和。此外,联合作用后听觉脑干诱发电位振幅下降非常明显。结果表明,顺铂与噪声联合作用对内耳的毒性有协同作用。     

香烟烟雾水溶液中的活性氧及其对姊妹染色单体互换的影响

SCE频率分析是研究环境诱变剂和致癌剂引起人类遗传物质损伤的有效手段。以人类外周血林巴细胞姊妹染色单体交换（SCE）为指标,评价了香烟烟雾水溶液的遗传毒性。首先,定量测定了香烟烟雾水溶液中含有H2O2、O2-等活性氧,从而间接证明了其潜在的遗传毒性。然后,将一定量的香烟烟雾水溶液加入人外周血淋巴细胞培养液中,结果表明实验组SCE频率明显高于空白对照组,因此直接证明了香烟烟雾水溶液的遗传毒性。     

四种重金属对中国鲎(Tachypleus tridentatus)胚胎发育的影响

分析了不同离子浓度的Zn、Pb、Cu、Cd对中国鲎胚胎发育的影响.结果表明①4种金属离子浓度≤渔业水域水质标准浓度4倍时,对中国鲎胚胎发育无明显毒性效应;≥渔业水域水质标准浓度8倍时,Cd离子和Cu离子对中国鲎胚胎发育无明显影响,而Zn离子组Pb离子组胚体发育随离子浓度提高而卵径变小,发育速度下降;②浓度为1.6mg/L时,Zn和Pb离子组胚胎致畸率分别为44%和58%,Pb离子组胚体孵化率仅2.67%;③4种重金属对中国鲎胚胎发育的毒性效应表现为Pb＞Zn＞Cu=Cd;④中国鲎胚胎对重金属有蓄积作用,蓄积效应随金属种类不同而异.     

贺普丁联合α-干扰素治疗慢性乙型肝炎的疗效观察

观察并总结了贺普丁联合α-干扰素治疗慢性乙型肝炎30例。结果 表明联合治疗组治疗12个月,HBV DNA阴转率高达86．67％,与单独贺普丁组63．3％相比,差异有显著性（P〈0．05）,与单独干扰素46．67％相比,差异亦有显著性（P〈0．01）。更加可喜的是联合治疗组治疗12个月时,HBeAg／Anti-HBe血清转换率达66．67％,而对照单独贺普丁组为43．33％,单独干扰素组为40．00％,经统计学分析均P〈0．05．差异有显著性。联合抗病毒治疗,YMDD的变异率明显较单独贺普丁治疗组低（P〈0．05）,而单独干扰素组治疗期间没有发现YMDD变异。     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

The early development of the tail and the transformation of the shape of the nucleus of the spermatid of the domestic fowl,Gallus gallus

Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices     

Characterization of the composition of the aqueous humor and the vitreous body of the eye of the frog Rana temporaria L

This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at ~ 1.5 × 10^− 4 mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.     

Observations on the permeability of the choriocapillaris of the eye

Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418