半胱胺对草鱼下丘脑-脑垂体组织共孵育中生长激素分泌的影响

采用静态孵育和放射免疫测定技术,研究了生长抑素抑制剂半胱胺盐酸盐对草鱼脑垂体组织单独孵育或下丘脑脑垂体组织共孵育中生长激素分泌的影响。结果表明：脑垂体组织单独孵育时,半胱胺盐酸盐(0．1、1和10mmol／L)对基础生长激素分泌无影响;而下丘脑脑垂体组织共孵育时,半胱胺盐酸盐(0．1、1和10mmol／L)对基础生长激素分泌有明显影响,且是剂量依存的。神经肽hGHRH、sGnRH—A和LHRH—A对CSH影响的下丘脑脑垂体组织共孵育中生长激素分泌均无协同作用。我们认为,半胱胺盐酸盐可在下丘脑水平调节生长激素释放,半胱胺盐酸盐调节草鱼离体生长激素分泌是由下丘脑途径介导的。     

绿原酸对草鱼鱼种生长、非特异性免疫和肉质的影响

为考察绿原酸对草鱼鱼种生长、非特异性免疫指标和肌肉品质的影响,在基础饲料中分别添加0.00%（对照组）、0.01%、0.02%、0.04%、0.06%、0.08%绿原酸,饲养平均体重（3.190.03） g的草鱼鱼种。经60d饲养,各组草鱼增重率分别为283.1%、305.7%、307.3%、329.6%、308.3%、303.0%,饲料系数分别为1.45、1.33、1.33、1.26、1.30、1.36;与对照组相比,添加0.04%绿原酸显著提高草鱼鱼种增重率,降低了饲料系数（P0.05）,添加0.01%、0.02%、0.06%绿原酸有提高草鱼鱼种增重率的趋势;在饲料中添加0.04%绿原酸可显著提高肝胰脏SOD活性（P0.05）,添加0.08%绿原酸可显著降低血浆天门冬氨酸氨基转移酶活性（P0.05）,各组肝胰脏溶菌酶、血浆碱性磷酸酶和丙氨酸氨基转移酶活性无显著差异（P0.05）。对肌肉成分的分析表明,各组水分、粗脂肪含量无显著差异（P0.05）,但添加0.04%绿原酸可显著提高肌肉粗蛋白、缬氨酸、异亮氨酸、组氨酸、必需氨基酸和总氨基酸含量（P0.05）。添加0.02%绿原酸可显著提高肌肉羟脯氨酸、胶原蛋白含量（P0.05）。综上所述,适量绿原酸能改善草鱼鱼种生长性能、非特异性免疫功能和肌肉品质。草鱼鱼种饲料中绿原酸添加量推荐为0.04%。       

鲤鱼（Cyprinus carpio）血清生长激素变化的酶联免疫吸附测定

本研究的目标是纯化鲤（Cyprinus carpio）生长激素,用于制备抗鲤鱼生长激素的单克隆抗体,建立鲤鱼生长激素的酶联免疫吸附测定（enzyme-linked immunosorbent assays,ELISA）技术,并用所建立的技术检测不同环境下养殖的鲤鱼的血清生长激素水平的变化．利用柱纯化技术纯化酵母表达草（Ctenopharyngodon idella）生长激素,免疫新西兰白兔（Oryctolagus cuniculus）,获得抗草鱼生长激素多克隆抗体．摘取鲤鱼垂体,从中提取生长激素,经层析纯化后,以变性聚丙烯凝胶电泳判断其分子量和纯度,并利用抗草鱼生长激素多克隆抗体以Western blot验证．纯化并经鉴定的鲤鱼生长激素作为免疫原被用于B淋巴细胞杂交瘤技术．共建立14株能够稳定分泌抗鲤鱼生长激素单克隆抗体的杂交瘤细胞系（FMU-cGH1-FMU-cGH14）,其中8个克隆（FMU．cGH1～FMU-cGH6,FMU-cGH12和FMU-cGH13）成功用于Western blot分析,9个克隆（FMU-cGHl-FMU-cGH7,FMU-cGH9和FMU-cGH10）可用于荧光标记和免疫组织化学分析．利用竞争性ELISA进行表位分析,结果表明这些单克隆抗体能够识别5个不同的表位．利用其中的FMU-cGH12作为包被抗体,FMU-cGH6作酶标抗体,建立了能够检测鲤鱼生长激素含量的ELISA技术．这一检测系统被证明具有高度的稳定性和灵敏度,能够检测并定量低至70pg／mL的生长激素．利用这一检测技术,发现限制进食和网箱养殖的鲤鱼其生长激素含量都有明显提高,分别为对照的6．9和5．8倍,显示不同生长环境下鲤鱼生长激素水平具有不同的反应．     

儿茶酚胺类药物对草鱼生长激素分泌的影响

本文研究了儿茶酚胺类药物对草鱼生长激素水平的影响。腹腔注射儿茶酚胺合成抑制剂α－甲基－对－酪氨酸（α－ＭＰＴ,５μｇ／ｇｂ·ｗ和５０μｇ／ｇｂ·ｗ）显著降低草鱼鱼种血清生长激素水平,腹腔注射儿茶酚胺能神经毒素６－羟多巴胺（６－ＨＯＤＡ,５μｇ／ｇｂ·ｗ和５０μｇ／ｇｂ·ｗ）也显著降低草鱼鱼种血清生长激素水平。阿卟吗啡（ＡＰＯ,７．１ｍｇ／Ｋｇｂ·ｗ和１２．５ｍｇ／Ｋｇｂ·ｗ）通过背大动脉插管注入草     

池塘饲养鱼类优化结构及其增产原理：Ⅰ.池塘越冬鱼种的生物学特性

1986—1989年的鱼种池塘越冬对比饲养与室内验证试验结果表明:长江以南草鱼种在冬季大部分时间仍能开口摄食。若不是密集越冬,并在晴天加强培育,越冬期间鱼种的平均生长率可达10—14％,鱼体丰满系数、血液中红细胞数、血红蛋白、血浆总蛋白含量,肌肉、肝脏中大部分营养成分较越冬前均无明显变化(p>0.05);补充投饵的半密集越冬组,鱼种体重、丰满系数能维持在越冬前水平(p>0.05),但血液、肌肉、肝脏中部分营养指标仍明显下降(p≤0.05);而饥饿条件下密集越冬组,鱼种消瘦9—14％,腹内肠系膜脂肪消耗殆尽,丰满系数、肝脏、肌肉中蛋白质、脂肪含量均明显下降(p≤0.05),且出现贫血状态。试验证明了越冬鱼种放养密度及饲养管理的不同,将影响鱼种质量的优劣和次年的成鱼养殖产量。为我国长江中下游大部,淮河、黄河中下游部分地区改革鱼种越冬制度,采取稀放精养的必要性和提高越冬鱼种质量与产量的可行性提供了理论根据。     

饥饿对月鳢几项血液指标的影响

对体长145．77±12．21cm,体重35．42±8．51g的月鳢（Channa asiatica）在正常摄食和饥饿4～20d状态下的几项血液指标进行了恻定。结果表明：饥饿组与对照组相比,随着饥饿时间的延长,月鳢的红细胞数量极显著减少（P〈0．01）;血红蛋白含量在前8d内是升高的,随后下降;血液中红细胞的渗透脆性有所上升,16d和20d差异显著（P〈0．05）;白细胞数量在整个饥饿期间一直呈上升趋势,在饥饿的第8d的测试中,白细胞数量的增加差异显著（P〈0．05）,12d以后的测试中白细胞数量的增加差异极显著（P〈0．01）。     

脆化草鱼(Ctenopharyngodon idellus C.et V)的病理生理生态学

分别就形态、生态和病理学性状对处于脆化终极阶段（LD50≥6．67kg／kgbw）的草鱼与普通草鱼进行比较分析。高度脆化的草鱼在生态学上出现明显变化,体长和体重的指数方程W=al^n显著不同于普通草鱼,DO阈值由平均值O．54mg／L上升到平均值1．68mg／L,CO2麻醉浓度（呼吸抑制值）由194mg／L高浓度下降到52．42～65．36mg／L,Root效应及Broh效应显著改变。解剖分析表明,草鱼脆化过程是渐进性组织病变的生理过程,包括各系统器官的功能性病变,最终使过度脆化的草鱼由于血液循环障碍直接导致死亡。草鱼脆化死亡具有明显的饲料（蚕豆）累积剂量——效应反应的毒理学规律,脆化草鱼的体长与体重的指数方程W=18．1719L^2.4360可作为华南地区商品脆肉鲩标准的参考。研究结果将为脆化草鱼规模化集约养殖和提高脆化草鱼长途运输成活率提供参考。     

大黄鱼生长激素基因的分离及序列测定

本文从大黄鱼（Pseucdosiaena crocea)脑下垂体中提取得到总RNA,根据近源鱼种生长激素（GH）基因保守序列设计了上游和下游引物,通过反转录和PC扩增得到大黄鱼生长激素cDNA片断,并进行序列分析。所得到的序列与近源鱼种生长激素基因序列有较高的同源性,断定为大黄鱼生长激素基因序列,本序列的测定对进一步研究其生物活性和工程菌中的表达及应用打下了基础。     

饥饿和再投喂对草鱼鱼种生物化学组成的影响

分析了饥饿１５天和再投喂２１天的草鱼鱼种肝脏和肌肉生物化学组成的变化，结果表明：（１）饥饿降低白肌ＲＮＡ／ＤＮＡ比值，蛋白质含量和肝脏ＲＮＡ／ＤＮＡ比值，使肝脏蛋白质含量升高，再投喂后，肝脏ＲＮＡ／ＤＮＡ比值，蛋白质含量和白蛋白质含量均恢复至正常投喂组水平，白肌ＲＮＡ／ＤＮＡ比值升高并显著高于正常投喂组水平；（２）饥饿状态下，肝脏和肌肉的脂类含量降低，水含量升高，再投喂后，肝脏和肌肉的脂类含量升高     

草鱼垂体生长激素分离纯化及其抗体制备的研究

应用碱性抽提,亲和层析,凝胶过滤和离子交换层析等技术从草鱼垂体中分离提纯出纯度高的草鱼生长激素（ｇｃＧＨ）用酶联免疫吸附测定法（ＥＬＩＳＡ）检测表明草鱼ＧＨ与大马哈鱼ＧＨ抗血清具有明显的免疫交叉反应,而与大马哈鱼催乳素（ＰＲＬ）抗血清没有交叉反应,聚丙烯酰胺凝胶电泳表明草鱼ＧＨ为单一蛋白带,其分子量约为２２５００道尔顿,等电聚焦揭示出草鱼ＧＨ主要由等电点为４．７和５．０的两条蛋白带组成,用纯化的草     

Studies of growth hormone secretion in juvenile diabetes.

To further investigate the GH secretion in juvenile diabetics, blood glucose (BG) and plasma growth hormone (GH) were determined during controlled exercise performed in basal condition and under glucose infusion, in 7 controls and 22 juvenile diabetics aged 12--35 years, 10 of them with fundal vascular lesions. In controls, glucose infusion significantly lowered the exercise induced GH rise observed under basal conditions. In diabetics, under basal conditions, diabetics with low basal BG (BG less than 100 mg/100ml) had higher GH secretion than those with high basal BG (BG greater than 140 mg/100 ml; p less than 0.05). Under glucose infusion, diabetics with normal BG peak values (not different from controls: BG = 284 +/- (SK) 45 mg/100 ml) had significantly higher plasma GH levels than controls (p less than 0.01). In contrast, in diabetics with BG peak value higher than controls (BG greater than 374 ng/100 ml), plasma GH levels were not different from control values. This study indicates that exercise induced GH secretion in diabetics is mainly related to actual BG levels. Furthermore, we found no relation between the magnitude of GH secretion and the presence of retinopathy in diabetics.     

Effects of starvation,re-feeding and timing of food supply on daily rhythm features of gut melatonin in carp (Catla catla)

Influences of starvation, re-feeding and time of food supply on daily rhythm features of melatonin (5-methoxy-N-acetyltryptamine) and its key regulator AANAT (arylalkylamine N-acetyltransferase) protein in the gut tissues were separately evaluated in carp Catla catla. The first experiment was aimed at demonstration of duration dependent effects of starvation and re-feeding after starvation on the daily profiles and rhythm features of gut melatonin and AANAT. Accordingly, juvenile carp were randomly distributed in three groups, which were (a) provided with balanced diet daily at a fixed time, that is, 10:00 clock hour or zeitgeber time (ZT) 4 (control), or (b) starved (for 2-, 4-, 6- or 8 days), or (c) initially starved for 8 days and then re-fed (for 2-, 4-, 6-, 8-, 12- or 16 days) daily with the same food and at the time (ZT4) used for control fish. The carp in each group were sampled for collection of gut tissues at six different time points at a regular interval of 4?h in a daily cycle. In another experiment, the influences of timing of food supply were separately examined in four fish groups, which were provided with a fixed amount of food once daily either at 06:00 or 12:00 or 18:00 or 24:00 clock hour corresponding to ZT0 or ZT6 or ZT12 or ZT18, respectively, for 7 days before sampling at 12 different time points with a regular interval of 2?h in a 24-h cycle. The study revealed a gradual increase in the mesor and amplitude values of melatonin and AANAT in gut with the progress of starvation till their values reached maximum at day-6 and remained steady thereafter. In contrast, re-feeding of 8-day starved fish resulted in a sharp decrease in their mesor and amplitude values after 2 days and then followed by a steady-state increase till re-attainment of their values close to control fish at the end of 16 days. The acrophase of these gut variables in each control, starved and re-fed fish was noted mostly at midday or ZT6. However, the results of another experiment demonstrated that a shift of food supply time led to a shift in their acrophase. The amount of residual food in the gut lumen in each, but not starved, fish by showing a significant positive correlation independently with the gut levels of melatonin and AANAT also indicated possible role of food as the synchronizer for their daily rhythms. Collectively, it appears reasonable to argue that daily profiles of gut melatonin and AANAT are strongly influenced by the availability of food, while their daily rhythm features seem to be dependent mostly on the time of food supply in carp.     

饥饿胁迫对草鱼鱼种非特异免疫力的影响

研究长期饥饿对草鱼(Ctenopharyngodon idellus)鱼种非特异性免疫力的影响。实验选取平均体质量(31.86?1.47g)的草鱼,随机分为2个实验组(对照组和饥饿组),每组三个平行,饥饿处理15、30、45和60d,测定饥饿对头肾和脾脏中自然杀伤(NK)细胞的杀伤活性、血清和肝胰脏中溶菌酶活性、血清中碱性磷酸酶活性的影响。结果表明：受饥饿胁迫的影响,草鱼鱼种自然杀伤性细胞在脾脏和头肾中的杀伤活性显著低于对照组(P<0.05)且不随着饥饿时间的延长发生显著性变化;随着饥饿时间的延长,血清和肝胰脏中溶菌酶呈现先升高后降低的趋势,血清碱性磷酸酶在饥饿15d、45d、60d时显著低于对照组;,饥饿组的碱性磷酸酶活性在饥饿30d以后,维持恒定。由此可见,长时间的饥饿胁迫降低了草鱼鱼种的免疫力。相比较而言,自然杀伤细胞的杀伤活性在评价鱼类的免疫状况时比溶菌酶和碱性磷酸酶更为灵敏。     

Effects of cysteamine--a somatostatin-inhibiting agent--on serum growth hormone levels and growth in juvenile grass carp (Ctenopharyngodon idellus)

Effects of cysteamine hydrochloride (CSH)-a somatostatin-inhibiting agent-on serum growth hormone (GH) levels and growth in juvenile grass carp (Ctenopharyngodon idellus) were studied. Intraperitoneal (i.p.) injection and single or 10-day feeding of different doses of CSH significantly increased serum GH levels. CSH and luteinizing hormone-releasing hormone analog (LHRH-A, D-Ala(6),Pro(9)-Net-LHRH), alone and in combination i.p. injection, and single or 10-day administration in diet resulted in an enhancement of serum GH contents; in addition, there was an additive, not synergistic effect of CSH and LHRH-A on elevation of serum GH levels. Ten day feeding of CSH, or CSH and LHRH-A, alone and in combination caused a significant increase in muscle RNA/DNA ratio. These results provide evidence that CSH significantly increases serum GH levels and promotes short-term growth in juvenile grass carp.     

个体大小对草鱼耐高温能力及升温过程中群体行为的影响

为考察个体大小对升温过程中草鱼的耐高温能力和群体行为的影响, 研究以草鱼(Ctenopharyngodon idellus)幼鱼为实验对象, 结果显示: 大个体群体的个体游泳速度明显大于小个体群体(P=0.039), 个体间距离和最近邻距离显著高于小个体群体(P<0.001); 大个体鱼群的极性与小个体群体并无差异。虽然大个体群体的致死高温与小个体群体无差异, 但前者的临界高温显著大于小个体群体(P<0.001)。升温过程会使鱼群个体游泳速度变快, 个体间距离和最近邻距离也相应增加, 但游泳速度同步性和群体极性维持不变。研究表明: 个体大小的增加可增强草鱼幼鱼对环境水温的耐高温能力; 升温不会影响不同个体大小草鱼幼鱼群体的协调性, 但会降低草鱼幼鱼的群体凝聚力。     

Primary-structural and evolutionary analyses of the growth-hormone gene from grass carp (Ctenopharyngodon idellus).

The growth-hormone (GH) gene of grass carp, one of the fastest-growing species of farmed fish, was isolated and the DNA sequenced. Only one GH gene is found in this species. This gene, which is 2.5 kb in length, has five exons and four introns, in common with all of the mammalian and the recently published common-carp GH genes. In the course of vertebrate evolution, the total lengths of the intron and the non-coding region of exon 5 of the GH gene have been shortened by 40-70%, whereas the encoding exons of the gene have been slightly increased. The more closely related species exhibit the closest sequence similarity in their GH genes. For example, the similarity of the exons is 84.1-93.2% between grass carp and common carp (within the same family of Syprinedae), 43.5-82.1% between grass carp and rainbow trout (in different orders of Teleostei) and 45.8-58.6% between grass carp and rat (in different grades of Vertebrata). In addition, similar DNA domains, such as thyroid-hormone-receptor-complex-binding site and cell-type-specific cis elements involved in regulation of expression of rat and human GH genes, have been localized in the corresponding regions of the grass-carp GH gene.     

青鱼生长激素的重组表达及其多克隆抗体的制备

以含有的青鱼生长激素编码区cDNA的重组质粒pbcGHc为模板,高保真PCR扩增青鱼生长激素（GH）成熟肽cDNA序列,定向插入原核表达载体pET-28a,构建青鱼GH原核表达质粒pET-bcGH。将pET-bcGH转化大肠杆菌BL21(DE3),IPTG诱导青鱼GH基因在大肠杆菌中的融合表达,SDS-PAGE凝胶电泳结果显示一条23 kDa的诱导表达重组青鱼GH带。以草鱼GH多克隆抗体为一抗,Western Blot证明,该重组青鱼GH具有免疫学活性。将经过亲和层析、透析纯化后的重组青鱼GH作为抗原,采用改进的方法对家兔进行皮下免疫注射,获得青鱼GH多克隆抗血清。以该多抗为一抗,Western Blot 可以检测出4 ng的抗原量;并且在青鱼垂体组织抽提液中和血清中检测到一种能与该抗血清作用的大小为21 kDa的蛋白质。这些结果表明本研究得到的青鱼GH多克隆抗血清具有较好的免疫特性。