双峰驼诱导排卵因子的生物学活性测定

公驼精清经ＤＥＡＥ－纤维素离子交换层析，共收集到７个蛋白组份，回归本动物，确定了可以诱导母驼排卵的组份，并经等电聚焦电泳加以证实。此外，分析了公驼精清的氨基酸组成，并对活性组份试行进一步纯化。     

双峰驼诱导排卵因子结构的研究

应用岛津紫外分光光度计系统地扫描并记录了公驼精清，各活性组分以及诱导排卵因子的紫外吸收光谱。扫描结果证明：公驼诱导排卵因子（OIF）在精清内相对稳定的原因，是OIF在精清内由多层分子量不同、性质各异的蛋白质所包裹，从而证明了我们以前的推测。     

双峰驼精清及其活性组分和馏分色氨酸含量的测定

应用岛津荧光分光光度计，测试了公驼精清、活性组、馏分以及诱导排卵因子的色氨酸含量。结果证明公驼精清和活性组，馏分含微量色氨酸；排卵因子是否含色氨酸详见讨论。     

双峰驼精液成分和品质的评价

应用紫外分光光度计扫描2峰精液品质和成分不同公驼精液的结果表明，正常精清出现19个蛋白和多肽组分，而品质不良的公驼精清仅为13个吸收峰；两者在碱性蛋白、酸性蛋白组分以及诱导排卵因子（OIF）的浓度和成分上，均存在明显的差别。     

双峰驼精清诱导排卵活性成分的DEAE—纤维素柱层析分离及其生物活性

选用DEAE－纤维素离子交换柱对双峰驼精清进行分离，各层析组分用于大鼠垂体组织体外培养，并给母驼肌注有活性的组分，以RIA法测定垂体组织培养液和母驼血浆中LH及FSH的浓度。结果表明，驼精清经DEAE－纤维素柱分离后可得到5个蛋白组分（L1－L5），其中L3在大鼠垂体培养时引起LH释放明显增加（P＜0．05），FSH则无明显变化。给卵巢上无大卵泡发育的母驼肌注有活性的组分L3，可引起血浆中LH明显升高，FSH也无变化。由此表明，L3在体内外试验中均具有引起LH释放的生物活性，它可能是诱导排卵的活性因子或其成分之一。     

双峰驼诱导排卵因子氨基酸组成的分析

分别测定了纯化的公驼精清诱导排卵因子的活性和固定部分肽类的小肽氨基酸组成,证明活性位序易降解,这一特性与所含的丝氨酸,谷氨酸,甘氨酸等有关。     

双峰驼排卵机制的研究——Ⅱ.精液的诱导排卵作用

双峰驼的排卵主要是由精液中的精清诱导发生的,阴道输入洗过的精子没有引起排卵。输精后是否排卵,和公驼精液的诱导作用及母驼个体的生理状况有关,排卵率约为86%。输精后的排卵时间与本交后的排卵时间一致。以2号公驼为例,精液1毫升能够诱导母驼排卵,但0.5毫升未引起排卵。精清中的诱导排卵物质能够耐受长期冰冻保存。公牛精液中也可能存在类似物质。肌肉注射 LH、LHRH 类似物及 HCG 能够引起排卵,但肌注及阴道输入 PGF_(2α)类似物后未排卵。     

双峰驼精清诱导排卵活性因子的阴离子交换柱Q Hyper D层析分离及生物活性鉴定

利用阴离子交换柱QHyperD对双峰驼精清进行分离 ,大鼠垂体组织培养及母驼肌注活性组分 ,RIA法测定垂体组织培养液和母驼血浆中LH及FSH的浓度。实验结果表明 ,驼精清经QHyperD柱层析后 ,共得到 6个组分 (F1～F6 ) ,其中F3和F5在大鼠垂体组织培养实验中具有生物活性 ,可引起LH的明显释放 (P <0 .0 5 )。在体实验时给卵巢上有成熟卵泡的母驼分别肌注F3或F5 ,肌注后 5～ 7h ,LH的释放也明显增加 (P <0 .0 5 )。F3可引起母驼发生排卵 ,F5则不能 ,说明F3可能就是驼精清中的诱导排卵活性因子或其成分之一。另外 ,无论是F3或是F5 ,在体内外实验中均未能引起FSH的分泌发生显著变化     

双峰驼精添诱导排卵活性因子的阴离子交换柱Q Hyper D层析分离及生物活性鉴定

利用阴离子交换柱Q Hyper D对双峰蛇精清进行分离，大鼠垂体组织培养及母驼肌注活性组分，RIA法测定垂体组织培养液和母驼血浆中LH及FSH的浓度。实验结果表明，驼精清经Q Hyper D柱层析后，共得到6个组分（F1－F6），其中F3和F5在大鼠垂体组织培养实验中具有生物活性，可引起LH的明显释放（P＜0.05）。在体实验时给卵巢上有成熟卵泡的母驼分别肌注F3或F5，肌注后5－7h，LH的释放也明显增加（P＜0.05）。F3可引起母驼发生排卵，F5则不能，说明F3可能就是驼精清中的诱导排卵活性因子或其成分之一。另外，无论是F3或是F5，在体内外实验中均未能引起FSH的分泌发生显著变化。     

双峰驼诱导排卵因子活性肽的分离

从双峰驼精清内分离出一种新的活性多肽，这种多肽是将公驼的精清经ＤＥＡＥ－纤维素和ＳｅｐｈａｃｒｙｌＳ－２００柱层析加以初步分离，在试验动物上测试，找到可以诱发母兔排卵的活性多肽，用十二烷基硫酸钠－聚丙烯酰胺凝胶电泳和等电聚焦电泳测定了这种多的分子量和等电点。     

Isolation of Ovulation-inducing Factors in the Seminal Plasma of Bactrian Camel (Camelus bactrianus) by DEAE-cellulose Chromatography

Ovulation in the Bactrian camel ( Camelus bactrianus ) depends upon the ovulation-inducing factor in the seminal plasma; however, little research has been conducted to isolate and identify the factor. The current study attempts to isolate and identify the bioactive fractions from the seminal plasma of Bactrian camel. The seminal plasma was fractionated by diethylamino-ethylcellulose (DEAE)-cellulose chromatography and five protein fractions were obtained. The bioactive of each fraction was estimated by rat pituitary tissue culture in vitro and by the intramuscular injection of the bioactive fraction to the female camels in vivo . The concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the pituitary culture media before and 6 h after the addition of each fraction and in the peripheral blood plasma collected from the camel immediately before and hourly after the injection of the active fraction were measured by radioimmunoassay. The results demonstrated that the third fraction (L3) had the bioactive potential to stimulate the release of LH in vitro from 11.82 ± 1.77 to 25.63 ± 3.84 mIU/ml after the addition of L3 to the culture media. The in vivo concentrations of LH in the blood plasma of the camel increased from 6.43 ± 0.14 before to 15.50 ± 2.64 ng/ml 6 h after injection of L3. However, the concentrations of FSH did not show any significant changes either in vitro or in vivo . The results clearly demonstrated the existence of LH-releasing associated fractions in the seminal plasma that appears to be separated by DEAE-cellulose matrix and the isolated L3 fraction might be the ovulation-inducing factor or one of its components.     

Separation and Purification of Ovulation-inducing Factors in the Seminal Plasma of the Bactrian Camel (Camelus bactrianus)

Ovulation in the Bactrian camel depends upon ovulation-inducing factors in the seminal plasma. The present study was conducted to isolate and purify the bioactive fractions from the seminal plasma of these camels. The seminal plasma was fractionated by anion-exchange chromatography, and six fractions were obtained. The bioactive potential of each fraction was estimated from its effect on rat pituitary tissue cultured in vitro and by the effect of an intramuscular injection of the fraction into female camels in vivo. Both the third fraction (F3) and the fifth fraction (F5) stimulated the release of LH in vitro and in vivo. In addition, female camels ovulated within 48 h after intramuscular injection of F3. However, neither F3 nor F5 had any significant effect on the secretion of FSH, either in vitro or in vivo. When F3 was further fractionated into four subfractions, the third subfraction (F3-3) still stimulated the in vitro release of LH, but not of FSH. An attempt to further purify the ovulation-inducing factors in F3-3 failed owing to the similarity of the molecular characters.     

哺乳动物精清诱导排卵活性蛋白质研究

在动物的精清里包含一些与动物生殖生理活动密切相关的活性蛋白和多肽。驼科动物的双峰驼、美洲驼、羊驼和澳洲考拉等哺乳动物精清中含有类似促性腺激素释放激素(GnRH)作用的活性蛋白质,诱导排卵因子(ovulation inducing factor,OIF),它能引发外周血液中促黄体素(LH)峰浓度的出现和诱发母畜排卵。OIF被认为是一种新的生物活性蛋白质或潜在的新型激素。甚至报道在牛等自发排卵动物以及人精清中也存在类似的活性蛋白。这类因子的发现对于哺乳动物生殖生理及繁殖的深入研究具有重要的意义,有助于动物克服繁殖障碍及繁殖疾病的治疗等。     

Effect of selection for litter size and body weight on hormone-induced ovulation rate in mice.

Genetic differences in natural vs hormone-induced ovulation rates were compared in immature female mice from five lines that had undergone long-term single-trait and antagonistic index selection for litter size and(or) 6-wk BW. Lines used were control (K); high litter size (L+); high BW (W+); low litter size and high BW (L-W+); and high litter size and low BW (L+W-). Natural ovulation rate at a mean age of 34.3 d and hormone-induced (5 IU of pregnant mare's serum gonadotropin followed 2 d later by 5 IU of human chorionic gonadotropin) superovulation rate at a fixed age of 31 d were obtained. Total number of eggs ovulated was affected by line (P less than .001), treatment (P less than .001), and line x treatment interaction (P less than .001). Line differences were subsequently tested within treatment because of the significant line x treatment interaction. Line differences were important (P less than .001) for natural ovulation, hormone-induced ovulation, and response to hormones. Mean natural ovulation rates for K, L+, W+, L-W+, and L+W- were 14.1, 19.8, 15.1, 13.6, and 16.4, respectively. Selection changed ovulation rate by 40, 16, 7, and -4% in the L+, L+W-, W+ and L-W+ lines, respectively (P less than .01). Hormone-induced ovulation rates in K, L+, W+, L-W+, and L+W- were 32.3, 24.6, 19.6, 20.9, and 22.1, respectively. Exogenous hormones increased ovulation by 18.2, 4.8, 4.6, 7.3, and 5.7 ova for K, L+, W+, L-W+, and L+W-, respectively (P less than .001). Lines with lower natural ovulation rates had higher responses to superovulation. Increased ovulation rate due to treatment ranged from 24.3% in L+ to 129% in K. These results indicate significant differences among lines in ovarian response to exogenous hormones.     

Serum protein electrophoretic pattern in young and adult camels

OBJECTIVE: To compare the electrophoretic pattern of serum proteins in clinically healthy adult camels (between 3 and 8 years of age) and camel calves (less than 3 months of age). DESIGN: Laboratory analysis of serum from healthy camels. PROCEDURE: Blood was collected from 30 healthy adult camels and 30 camel calves and the serum separated. Total protein of each serum sample was estimated by automated chemistry analyser. The proteins were fractionated by automated electrophoresis on agarose gel. RESULTS: Serum proteins migrated on the agarose gel as one albumin, two alpha (alpha1 and alpha2-globulins), two beta (beta1 and beta2-globulins) and one gamma-globulin fractions. In adult camels the mean concentration of total protein, albumin alpha1, alpha2, beta1, beta2 and gamma-globulins was 56.8 +/- 1.5, 30.7 +/- 0.8, 2.4 +/- 0.1, 3.2 +/- 0.1, 9.7 +/- 0.3, 3.4 +/- 0.2 and 8.6 +/- 0.3 g/L, respectively. These values in calves were 49.7 +/- 1.8, 23.7 +/- 0.8, 3.2 +/- 0.2, 3.1 +/- 0.2, 14.2 +/- 0.2, 4.0 +/- 0.2 and 4.1 +/- 0.2 g/L, respectively. CONCLUSION: The concentration of total proteins, albumin and gamma-globulins was higher (P < 0.05) in the adult camels than in camel calves. The concentrations of beta1 globulins was higher (P < 0.05) in calves as compared to adult camels.     

双峰驼诱导排卵机理的研究Ⅲ--双峰驼诱导排卵因子分布与代谢途径的研究

从公驼精清内经DEAE－DE52柱上分离的诱志排卵活性蛋白（Ovulation-inducing bioactive protein,OIP)应用^125I标记后输入母驼阴道，通过血液循环到达垂体。经测定，OIP遍布母驼体内各脏器，其中以垂体、肝脏、心脏、舌肌等浓度较高，而在眼角膜、耳尖部以及被毛中含量很少。消除半衰期长达72.6h，其代谢主要经肝胆排泄，也有部分活性蛋白通过泌尿、生殖道排泄。     

Leishmanicidal activity in vitro of Musa paradisiaca L. and Spondias mombin L. fractions

Visceral leishmaniasis (VL) is a zoonotic disease characterized by infection of mononuclear phagocytes by Leishmania chagasi. The primary vector is Lutzomyia longipalpis and the dog is the main domestic reservoir. The control and current treatment of dogs using synthetic drugs have not shown effectiveness in reducing the incidence of disease in man. In attempt to find new compounds with leishmanicidal action, plant secondary metabolites have been studied in search of treatments of VL. This study aimed to evaluate the leishmanicidal activity of Musa paradisiaca (banana tree) and Spondias mombin (cajazeira) chemical constituents on promastigotes and amastigotes of L. chagasi. Phytochemical analysis by column chromatography was performed on ethanol extracts of two plants and fractions were isolated. Thin layer chromatography was used to compare the fractions and for isolation the substances to be used in vitro tests. The in vitro tests on promastigotes of L. chagasi used the MTT colorimetric method and the method of ELISA in situ was used against amastigotes besides the cytotoxicity in RAW 264.7 cells. Of the eight fractions tested, Sm1 and Sm2 from S. mombin had no action against promastigotes, but had good activity against amastigotes. The fractions Mp1 e Mp4 of M. paradisiaca were very cytotoxic to RAW 264.7 cells. The best result was obtained with the fraction Sm3 from S. mombin with IC(50) of 11.26 μg/ml against promastigotes and amastigotes of 0.27 μg/ml. The fraction Sm3 characterized as tannic acid showed the best results against both forms of Leishmania being a good candidate for evaluation in in vivo tests.     

Sero-prevalence of agglutinins to Listeria monocytogenes in Nigerian domestic animals

A survey using tube agglutination test was conducted to determine the antibody prevalence to Listeria monocytogenes serotypes 1/2a, 1/2b, 1/2c, 3a and 4b in 1,190 serum samples of 8 animals species from various sources in Kano and Kaduna states of Nigeria. Following absorption with Staphylococcus aureus antigen to remove cross-reacting agglutinins, 52 (68.4 p. 100) of the horse samples were positive. Twenty-six (36.1 p. 100) pig, 52 (20.8 p. 100) cattle, 50 (20.0 p. 100) goat, 20 (20.0 p. 100) dog, serum samples were also positive. Free-ranging chickens had an antibody prevalence of 18 (32.1 p. 100) while those intensively managed had 3 (6.8 p. 100), a difference found to be statistically significant (P less than or equal to 0.01; X2). Sheep sera collected from Zaria abattoir had a prevalence of 30 (14.7 p. 100) while those from Ahmadu Bello University, Veterinary Teaching Hospital had 6 (13.0 p. 100) prevalence. The prevalence in camel was 4 (4.3 p. 100). Overall, of the 1,190 serum samples tested, 26 (21.9 p. 100) were sero-positive for L. monocytogenes agglutinins. Each species of animal tested for L. monocytogenes was positive for all five serotypes, except camel which was negative for serotype 3a. Fourty-four (53.0 p. 100) samples were positive at a titre of greater than or equal to 480 for serotypes 1/2a, 60 (58.3 p. 100) for 1/2b, 57 (52.3 p. 100) for 1/2c, 7 (13.7 p. 100) for 3a and 23 (39.0 p. 100) for 4b. It is concluded that L. monocytogenes infection is widespread in domestic animals in Nigeria.     

益生菌发酵驼乳对慢性肾功能衰竭的治疗作用

目的：比较不同菌种发酵的驼乳制品对腺嘌呤所致大鼠慢性。肾功能衰竭（CRF）的缓解作用。方法：采用腺嘌呤复制CRF大鼠模型,以不同发酵剂发酵的驼乳作为受试物进行灌胃干预。通过检测大鼠的饮食饮水情况、排尿量、尿液和血清常规指标及肾脏病理组织学变化,评估各发酵驼乳对CRF大鼠的治疗效果。结果：发酵驼乳均可改善肾功能衰竭大鼠的一般生理状况,可降低大鼠血肌酐（Scr）、尿素氦（BUN）水平,减缓尿蛋白（UP）,调节Ca、P的含量,提高过氧化物歧化酶（SOD）、血清总蛋白（STP）水平,具有保护肾功能的作用,其中LIcaseiZhang发酵的驼乳对CRF大鼠的改善效果最佳。     

Isolation and partial characterization of excretory/secretory antigens of Gastrothylax crumenifer

The present study was carried out to identify the excretory/secretory (E/S) antigens of the rumen infecting digenetic trematode Gastrothylax crumenifer that may be useful for the immunodiagnosis of rumen amphistomosis particularly during the pre-monsoon season during which this rumen parasite stops shedding eggs. The in vitro released E/S proteins were purified on a Sephadex G-200 column. The gel filtration profile revealed three distinct fractions F1-F3 where F1 and F3 appeared as sharp peaks while the F2 fraction was dispersed. The antibody titre against each of the purified E/S fractions was determined by ELISA using anti-whole E/S polyclonal antibodies raised in rabbit. Among the three fractions, the antibody titre against F1 was highest (1:12,800) whereas IgG titre was very low (1:50) for fraction F2 and F3 (1:100). Of the total polypeptides resolved on gradient SDS-PAGE, only a few antigenic polypeptides were detected in each fraction with hyperimmune anti-serum as revealed by Western Blot analysis. However, a 33 kDa antigen detected in each fraction appeared to be immunodominant which could be exploited for the diagnosis of the pouched amphistome.