东北虎幼体血清蛋白及胆汁酸的测定与分析

用HITACHI 7600-20全自动生化分析仪对13只东北虎幼体进行了总蛋白、白蛋白、球蛋白、前白蛋白及胆汁酸5项生化指标的测定,实验数据经SPSS软件统计分析.其结果为:总蛋白含量66.06±6.80 g/L,白蛋白含量34.19±2.51 g/L,球蛋白含量31.89±4.46 g/L,前白蛋白含量13.22±11.64 g/L,胆汁酸含量4.48±3.89 μmol/L.东北虎幼体雌雄个体间5项生化指标均无显著差异(P0.05).东北虎幼体与成体间,总蛋白、白蛋白、球蛋白含量均呈极显著差异(P<0.01).东北虎幼体与华南虎幼体间,总蛋白、球蛋白含量呈显著差异(P<0.05);白蛋白含量无显著差异(P0.05).     

东北虎幼体消化系统蛋白水解酶的初步研究

蛋白水解酶在许多生命活动中是必需的物质(Vassalli and Pepper,1994)。蛋白质的酶解修饰(Xuet al.,1999)、细胞迁移、组织再生与修复、消化系统对蛋白质的消化等均与蛋白水解酶有关(Baimbridgeet al.,1992),且蛋白水解酶功能失调会导致许多疾病(Teichertet al.,1989)。东北虎(     

东北虎血液成分的测定与分析

用氰化高铁血红蛋白法、改良纽鲍氏计算板法、显微测量法、低渗ＮａＣｌ试管法、微量毛细吸管离心法、血细胞计算板法和瑞特氏染色三区计数法,分别测得和算出８只东北虎血液中血红蛋白含量１３１±７．５ｇ／Ｌ、红细胞数７．１１±Ｏ．５３×１０１２／Ｌ、红细胞直径５．５８（４．６２—６．５５）μｍ、红细胞渗透脆性０．６０６±０．０６５—０．４３０±０．０４５％、红细胞压积容量３８．１±２．６８％、白细胞总数２３．３±７．５×１０９／Ｌ、嗜中性白细胞５７．９％、淋巴细胞３５．６％、单核细胞３．５％、嗜酸性粒细胞１．８％、嗜硷性粒细胞＜１％、平均红细胞体积５３．６±２．７５ｆＬ、平均红细胞血红蛋白含量１７．９±１．２５Ｐｇ、平均每个红细胞血红蛋白分子数１５８９±１１０×１０５个、平均红细胞血红蛋白浓度３３．５±２．７％;用全自动凯氏定氮法测定并计算得出血清总蛋白含量７３．６±１２．６ｇ／Ｌ、血清白蛋白含量４６．５±３．２％及血清球蛋白含量２７．１±１３．１ｇ／Ｌ。上述各项结果为保护东北虎提供了难得而有意义的生理参考值。     

吉林省东北虎的调查

经过8年的调查研究,确定了东北虎在吉林省的分布区有3处,数量为7－9只。即大龙岭分布区3－5只,哈尔巴岭分布区1只,张广才岭分布区3只。东北虎的适宜生境为海拔800－1100m的中低山：人口压力在15人／km^2以下,被捕食动物野猪、狍、马鹿的密度在2．5只／km^2以上。林型为以柞木为主的阔叶混交林,林龄为中成林。     

4种生态因子对虎斑乌贼幼体生长与存活的影响

旨在研究温度、盐度、饵料种类、投饵量对虎斑乌贼（Sepia pharaonis）幼体生长与存活率的影响,以确定其生长发育的最佳生态条件,为人工育苗提供理论依据。在室内控制条件下,采用单因子试验研究了不同温度（18、21、24、27、30和33℃）、不同盐度（18、21、24、27、30和33）、不同饵料种类（虾糜、活糠虾、卤虫无节幼体、桡足类、死糠虾、虾糜＋强化卤虫后无节幼体、强化卤虫后无节幼体）、活糠虾不同投饵量（0、2、4、6、8和10 g/d）对虎斑乌贼幼体生长发育的影响。结果显示：不同温度对虎斑乌贼幼体生长影响显著（P〈0.05）,最适温度为24-27℃,在最适温度下,存活率为84.4%-91.1%,特定生长率为4.82%-6.13%,存活率（y）与温度（x）的函数关系为y=-0.15x3＋30.637x-447.002（r2=0.923）。幼体适宜盐度为24-33,最适盐度为27,在最适盐度条件下,其存活率为（90.0±5.29）%,特定生长率为（3.71±0.34）%。投喂7种开口饵料,以活糠虾效果最佳,存活率为（96.7±2.94）%,特定生长率为（3.77±0.23）%;强化卤虫后无节幼体效果次之,存活率为（95.6±2.31）%,特定生长率为（2.54±0.15）%,其余各组培养效果均不理想。投喂活糠虾,随着投饵量的增加,其存活率随之提高,摄食量（y）与个体重（x）的回归方程为y=0.227x-0.063（r2=0.921）。     

东北虎中SOX基因的检测

应用特异于HMG bOX区域的兼并引物 ,扩增了东北虎的SOX基因。在扩增产物中发现五条大小分别为 2 2 0 ,2 70 ,3 5 0 ,4 3 0和 5 60bp的扩增带。经过与地高辛标记的人SRY基因进行Southern杂交表明这五条扩增带均呈现阳性 ,说明它们均为东北虎的SOX基因片段 ,这些基因保守区的长度在基因组DNA水平上存在着明显的差异。     

东北虎雌性生殖系统的组织学

对10月份意外死亡的2例东北虎（Panthera tigris altaica)雌性生殖系统进行了组织学研究。卵巢基质中分布着一定数量的有腔卵泡及闭锁卵泡。间质腺不发达,在闭锁卵泡上皮样细胞增殖,形成一定量的间质腺细胞。输卵管漏半斗部至峡部黏膜上皮由柱状逐渐转化炎立方,黏膜形成许多皱褶,皱褶以漏斗部和壶腹部最多,子宫内膜上皮为单层立方上皮。6岁个体子宫腺上皮为单层柱状上皮;2岁个体在子宫角处的子宫腺上皮为单层柱状上皮,在子宫体外多为单层立方上皮,子宫腺密度从卵巢端到子宫体端呈阶梯下降,阴道黏膜上皮大多为2－3层的复层扁平上皮。     

东北虎,云豹,金猫红细胞上C3b受体的测定

本文通过对健康的东北虎、云豹和金猫红细胞上C_(3b)受体和免疫复合物的检测,证实这三种动物的红细胞,除了具有免疫粘附,形成免疫复合物的花环(其花环率分别为7.00、6.50和5.67)外,还可通过其膜上的C_(3b)受体结合免疫复合物,形成C_(3b)受体的花环,其花环率分别为11.50、11.88和11.83。表明东北虎、云豹和金猫的红细胞具有较强的免疫功能。     

笼养东北虎行为的时间分配

1998 年4 月至2001 年3 月, 采用瞬时扫描取样法和全事件取样法对哈尔滨动物园的4 只笼养东北虎的活动规律进行了研究。结果表明, 笼养东北虎在全年的昼夜行为时间分配表现为睡眠所占比值最多; 其次是卧息和活动; 摄食和其它行为(包括饮水、排尿、排粪、修饰、嗅闻、嬉戏、站立、直立、发声等) 最少。在不同季节笼养东北虎的活动变化规律基本相似, 一昼夜的睡眠、运动和卧息均有两个高峰期, 但高峰期出现和持续的时间有所差异。睡眠的高峰期在10 : 00～14 : 00 和20 : 00 至次日06 : 00 , 活动的高峰期在05 : 00～10 : 00 和14 : 00～17 : 00 , 卧息的高峰期在05 : 00～10 : 00 和13 : 00～20 : 00 , 摄食仅在16 : 00～20 : 00 有一个高峰期。冬季(12～2 月) 与春(3～5 月) 、夏(6～8 月) 、秋季(9～11 月) 相比, 东北虎在白昼睡眠的时间显著减少,夜晚则显著增多。     

调控精子获能的相关因素

获能是精子发生顶体反应以及与卵子结合之前所必需的生理过程,目前精子获能的机制得到初步阐明,获能伴随着质膜重组,离子通道的调节,胆固醇的流失以及许多蛋白磷酸化状态的改变.获能同时受到内在和外在因子的调节,其中胆固醇、HCO3-、Ca2+以及蛋白磷酸化在精子获能过程中发挥着重要作用.     

Contribution of anion transporters to the acidosis-induced swelling and intracellular acidification of glial cells

This study examines the contribution of anion transporters to the swelling and intracellular acidification of glial cells from an extracellular lactacidosis, a condition well-known to accompany cerebral ischemia and traumatic brain injury. Suspended C6 glioma cells were exposed to lactacidosis in physiological or anion-depleted media, and different anion transport inhibitors were applied. Changes in cell volume and intracellular pH (pH(i)) were simultaneously quantified by flow cytometry. Extracellular lactacidosis (pH 6.2) led to an increase in cell volume to 125.1 +/- 2.5% of baseline within 60 min, whereas the pH(i) dropped from the physiological value of 7.13 +/- 0.05 to 6.32 +/- 0.03. Suspension in Cl(-)-free or HCO(3)(-)/CO(2)-free media or application of anion transport inhibitors [0.1 mM bumetanide or 0.5 mM 4, 4'-diisothio-cyanatostilbene-2,2'-disulfonic acid (DIDS)] did not affect cell volume during baseline conditions but significantly reduced cell swelling from lactacidosis. In addition, the Cl(-)-free or HCO(3)(-)/CO(2)-free media and DIDS attenuated intracellular acidosis on extracellular acidification. From these findings it is concluded that besides the known activation of the Na(+)/H(+) exchanger, activation of the Na(+)-independent Cl(-)/HCO(3)(-) exchanger and the Na(+)-K(+)-Cl(-) cotransporter contributes to acidosis-induced glial swelling and the intracellular acidification. Inhibition of these processes may be of interest for future strategies in the treatment of cytotoxic brain edema from cerebral ischemia or traumatic brain injury.     

不同来源小球藻对岩溶水Ca2+、HCO3-利用的初步研究

以外源小球藻和岩溶区筛选出的土著小球藻为研究对象, 在封闭体系中比较研究了两种不同来源小球藻对典型岩溶水中Ca2+、HCO3-的利用、藻细胞数量与其对Ca2+、HCO3-的利用率的关系和体系pH的变化。结果表明, 土著小球藻利用Ca2+、HCO3-的能力强于外源小球藻, 但外源小球藻对Ca2+的利用量高于土著小球藻, 而二者对HCO3-的利用量相同, 并且外源小球藻能够以胞外CaCO3形式产生沉淀, 而土著藻则不能形成沉淀。其次两体系中pH的变化显示, 两种小球藻光合作用都是先以水体中CO2为光合作用碳源, 然后利用HCO3-。外源小球藻能将岩溶水中29.648%的HCO3-吸收, 而土著藻能将40.625%的HCO3-通过其在食物链中的初级生产地位将岩溶碳汇转化进入到生态系统, 表现为净碳汇效应。     

Determination of glucose,urea and penicillin using enzyme-pH-electrodes

Conventional hydrogen ion glas electrodes have been used for the preparation of enzyme-pH-electrodes by either entrapping the enzymes within polyacrylamide gels around the electrode or as liquid layer trapped within a cellophane membrane. The enzymes were glucose oxidase, urase and penicillinase.The pH response to glucose concentration was about linear within 10^?1–10^?3 M glucose and for urea linear within 5·10^t—–5·10^?5M. The pH response to penicillin was about linear in the range from 10^?3–10^?2 M resulting in a pH shift of 1.4 units; reproduceable pH response was obtained down to concentrations of 3·10^?5 M.Studies as to the effect of buffer using an urease–pH-electrode showed a buffer concentration of 10^?2 M a substantial shift of about one pH-unit in the range of 10^?4 to 10^?2 M urea. Both urease- and penicillinase–pH-electrodes were tested as to stability showing no decrease in pH response except at high substrate concentration (1·10^?2 M) over a period of 2–3 weeks kept at room temperature.     

Relative contribution of the Na(+)/Ca(2+) exchanger, mitochondria and endoplasmic reticulum in the regulation of cytosolic Ca(2+) and catecholamine secretion of bovine adrenal chromaffin cells

The relative importance of mitochondria, the Na(+)/Ca(2+) exchanger (NCX) and the endoplasmic reticulum (ER) in the regulation of the cytosolic Ca(2+) concentration ([Ca(2+)](i)) were examined in bovine chromaffin cells using fura-2 for average [Ca(2+)](i) and amperometry for secretory activity, which reflects the local Ca(2+) concentration near the exocytotic sites. Chromaffin cells were stimulated by a high concentration of K(+) when the three Ca(2+) removal mechanisms were individually or simultaneously inhibited. When the mitochondrial Ca(2+) uptake was inhibited, the [Ca(2+)](i) decayed at a significantly slower rate and the secretory activity was higher than the control cells. The NCX appears to function only in the initial phase of [Ca(2+)](i) decay and when the ER Ca(2+) pump is blocked. Similarly, the ER had a significant effect on the [Ca(2+)](i) decay and on the secretion only when the NCX was blocked. Inhibition of all three mechanisms leads to a substantial delay in [Ca(2+)](i) recovery and an increase in the secretion. The results suggest that the three mechanisms work together in the regulation of the Ca(2+) near the Ca(2+) channels and exocytotic sites and therefore modulate the secretory activity. When Ca(2+) diffuses away from the exocytotic sites, the mitochondrial Ca(2+) uptake becomes the dominant mechanism.     

Effects of Monovalent and Divalent Cations on Ca2+ Fluxes Across Chromaffin Secretory Membrane Vesicles

Abstract: Bovine chromaffin secretory vesicle ghosts loaded with Na⁺ were found to take up Ca²⁺ when incubated in K⁺ media or in sucrose media containing micromolar concentrations of free Ca²⁺. Li⁺- or choline⁺loaded ghosts did not take up Ca²⁺. The Ca²⁺ accumulated by Na⁺-loaded ghosts could be released by the Ca²⁺ ionophore A23187, but not by EGTA. Ca²⁺ uptake was inhibited by external Sr²⁺, Na ⁺, Li ⁺, or choline ⁺. All the ⁴⁵Ca²⁺ accumulated by Na⁺-dependent Ca²⁺ uptake could be released by external Na ⁺, indicating that both Ca²⁺ influx and efflux occur in a Na⁺-dependent manner. Na ⁺ -dependent Ca²⁺ uptake and release were only slightly inhibited by Mg²⁺. In the presence of the Na⁺ ionophore Monensin the Ca²⁺ uptake by Na ⁺-loaded ghosts was reduced. Ca²⁺ sequestered by the Na⁺-dependent mechanism could also be released by external Ca²⁺ or Sr²⁺ but not by Mg²⁺, indicating the presence of a Ca²⁺/Ca²⁺ exchange activity in secretory membrane vesicles. This Ca²⁺/Ca²⁺ exchange system is inhibited by Mg²⁺, but not by Sr²⁺. The Na ⁺ -dependent Ca²⁺ uptake system in the presence of Mg²⁺ is a saturable process with an apparent K_m of 0.28 μM and a V_max= 14.5 nmol min^?1 mg protein^?1. Ruthenium red inhibited neither the Na⁺/Ca²⁺ nor the Ca²⁺/Ca²⁺ exchange, even at high concentrations.