海带,裙带菜和紫菜蛋白酶谱的研究

采用复性电泳方法对海带（ＬａｍｉｎａｒｉａＬａｍｘ．）、裙带菜（ＵｎｄａｒｉａＳｕｒｉｎｇａｒ）和紫菜（ＰｏｒｐｈｙｒａＣ．Ａｇ．）３个属１１种海藻的蛋白酶进行了研究,分析了海带、裙带菜和紫菜在不同ｐＨ条件下,蛋白酶种类的分布情况和活性差异。结果表明：（１）蛋白酶种类和活性受ｐＨ的影响和制约,其最适ｐＨ为８．０,此条件下蛋白酶的种类最多,活性最强;（２）亲缘关系越近,蛋白酶种类和活性越相似,海带同属不同种之间存在相同的蛋白酶,而不同属的海藻中很难找到相同的蛋白酶带;（３）海带属不同种所共有的两种蛋白酶（分子量１９ｋＤ和１８ｋＤ）是其特征性酶带;（４）蛋白酶谱可作为海藻分类依据之一。     

月鳢消化器官指数及蛋白酶活性对饥饿的响应

经实验发现饥饿处理对月鳢各消化器官中蛋白酶活性有显著的影响,测定了实验组和对照组中月鳢各消化器官的蛋白酶活性的变化后,经统计分析,结果显示,在饥饿了4、8、12、16、20d后,胃、肠、肝胰脏中蛋白酶的活性与对照组相比较都出现了极显著变化（p〈0.01）;比胃重、比肝胰脏重以及比肠重在饥饿处理了4d和8d后并未出现显著变化（p〉0.05）,但在饥饿处理时间超过8d后,比肠重与对照组相比较出现了显著变化（p〈0.05）。     

云斑尖塘鳢消化酶活力的研究

本文采用酶学分析方法研究了云斑尖塘鳢在正常摄食状态与饥饿的状态下胃、肠及肝胰脏组织中蛋白酶、淀粉酶和脂肪酶的活性。结果显示,在30℃的条件下,正常摄食组样本在酸性条件下的蛋白酶活力表现为:胃后肠肝胰脏前肠,中性和碱性条件下:后肠肝胰脏前肠及胃;饥饿组样本仅有胃表现出较高的酸性蛋白酶活性,其他器官的蛋白酶活性均很低。在正常和饥饿实验组中肝胰脏的淀粉酶活性均高于其他器官,胃肠的淀粉酶活性均较低。正常摄食组中脂肪酶活力后肠肝胰脏;而在饥饿组中仅有肝胰脏检测到脂肪酶活性。结果表明,云斑尖塘鳢适度饥饿组较正常摄食组消化酶活性大幅降低;其高蛋白酶活力及中等脂肪酶活力与其肉食性相一致;此外云斑尖塘鳢也具备少量的淀粉消化能力。     

大鵟消化系统蛋白水解酶种类和活性分析

采用蛋白水解酶复性电泳(G-PAGE)技术对大(Buteo hemilasius)消化系统5种器官腺胃、胰脏、十二指肠、空肠、大肠蛋白水解酶的种类和性质进行了研究,以期为研究野生鸟类的分类地位、系统演化提供基础资料,结果表明,①受pH值的影响和制约,大消化系统蛋白水解酶的活性在碱性、中性与酸性条件下递减;②在酸性条件下,45 ku蛋白水解酶存在于除腺胃外的各受检器官;③pH 7.0时,腺胃、胰脏酶谱相似,均含有683、5、342、0 ku的蛋白水解酶;④pH 8.0时,空肠和十二指肠的蛋白水解酶种类最多、活性最强,分别检出8种和7种蛋白水解酶。总之,pH值对蛋白水解酶的活性有明显的制约作用,46、41ku蛋白水解酶随着pH值的增高而失去活性,为酸性蛋白水解酶,250、2064、5 ku蛋白水解酶随着pH值的增高活性逐渐增强,为碱性蛋白水解酶。十二指肠和空肠的蛋白水解酶种类多、活性强,可能为蛋白质消化的主要场所。     

球孢白僵菌凝乳弹性蛋白酶(BbPr1)的纯化与特性*

以几丁质为底物,加入基本盐培养基中,诱导球孢白僵菌（Beauveriabassiana）产生分解昆虫表皮的蛋白酶。诱导物中,蝉蜕诱导的蛋白酶总活性、比活较高,经超滤、离子交换层析、亲和层析、制备性IEF电洗脱纯化了一种有凝乳弹性蛋白酶（Pr1）活性的蛋白酶BbPr1。经SDS－PAGE电泳银染后是单带,HPLC凝胶过滤显示单峰。BbPr1为单体酶,分子量为33.6kD左右,pI为7.4。底物专一性测定显示,BbPr1能水解Phe或Leu形成的酸胺键和肽键。BbPrl可被PMSF抑制,表明其活性中心有Ser残基;BbPr1还可被胰凝乳蛋白酶抑制剂TPCK和凝乳弹性蛋白酶抑制剂TEI等所抑制;胰蛋白酶抑制剂beupeptin和Epianstatin,及胃蛋白酶抑制剂Pepstain对BbPr1活性无影响。还研究了BbPr1的最适作用pH和pH稳定性。     

越鞠保和丸联合穴位埋线对肝胃不和型功能性消化不良患者胃动力指标、血清胃肠激素和胃蛋白酶原的影响

目的：探讨越鞠保和丸联合穴位埋线对肝胃不和型功能性消化不良（FD）患者胃动力指标、血清胃肠激素和胃蛋白酶原的影响。方法：采用随机数字表法将2020年6月～2023年6月期间北京市鼓楼中医医院收治的123例FD患者分为对照组（61例）和研究组（62例）。对照组接受常规治疗联合穴位埋线治疗,研究组在对照组的基础上结合越鞠保和丸治疗。对比两组疗效、中医证候积分、胃动力指标、胃肠激素指标和胃蛋白酶原指标。结果：研究组的临床总有效率高于对照组（P<0.05）。治疗后研究组主症、次症、中医证候积分总分低于对照组（P<0.05）。治疗后研究组胃动素（MLT）高于对照组,生长抑素（SS）、胃泌素（GAS）低于对照组（P<0.05）。治疗后研究组餐后胃窦收缩频率（F）、收缩幅度（P）、运动指数（MI）高于对照组（P<0.05）。治疗后研究组胃蛋白酶原Ⅰ（PGⅠ）、胃蛋白酶原Ⅱ（PGⅡ）高于对照组（P<0.05）。结论：越鞠保和丸联合穴位埋线可有效改善肝胃不和型FD患者的血清胃肠激素和胃蛋白酶原水平,促进胃动力,进而改善胃肠功能。     

竹叶青蛇消化道内分泌细胞的免疫组织化学定位

应用8种特异性胃肠激素抗血清对竹叶青蛇（Trimersurus stejnegeri）消化道内分泌细胞进行了免疫组织化学定位。5－羟色胺细胞和生长抑素细胞在整个消化道中均有分布。5－羟色胺细胞以十二指肠和回肠密度为最高,空肠、直肠和幽门其次,胃体较低,食道和贲门中偶见;生长抑素细胞在十二指肠处数量最多,胃体、幽门和空肠其次,其余消化道各段偶见;胰高血糖素细胞只见于十二指肠和空肠;血管活性肠肽在贲门和胃体处有少量阳性细胞分布,食道中偶见。以上各处内分泌细胞的分布密度差异显著。未检出胃泌素、P－物质、胰多肽和胰岛素免疫反应阳性细胞。上述内分泌细胞的分布特点可能与竹叶青的摄食习性和生活环境有关。     

扬子鳄小胃黏膜的组织化学和细胞学

扬子鳄（Alligator sinensis）的小胃是在胃幽门部与十二指肠的交接处由胃幽门括约肌和十二指肠括约肌突入管腔形成的一个小腔,其生理功能一直不清楚。本文采用组织化学、电子显微镜和免疫组织化学技术对扬子鳄小胃黏膜的组织化学成分、细胞超微结构及细胞类型进行了较全面的研究。小胃黏膜上皮PAS反应呈强阳性,AB染色呈弱阳性,主要分泌中性糖蛋白和少量的含硫酸性糖蛋白。电子显微镜下,扬子鳄小胃黏膜上皮主要由表面黏液细胞组成,偶见内分泌细胞。小胃腺部则70％-90％为内分泌细胞,其余为少量的腺黏液细胞和泌酸胃酶细胞。应用7种胃肠激素的抗血清在小胃黏膜中检测出了5-羟色胺（5-HT）、胃泌素（Gas）、胰高血糖素（Glu）、生长抑素（SS）、P-物质（SP）和血管活性肠肽（VIP）细胞,以Glu细胞密度最高,VIP细胞密度最低。未检测到胰多肽（PP）细胞。本研究结果表明,从组织化学成分和细胞类型看,扬子鳄的小胃与胃同源;从细胞超微结构和内分泌细胞所占百分比例看,扬子鳄的小胃已出现了明显的特化,泌酸胃酶细胞中未见泌酸小管,可能没有泌酸功能。内分泌细胞含量丰富,可能在调节胃肠道功能中发挥重要作用[动物学报54（6）：1044—1050,2008]。     

有机碳和无机碳对3种真菌胞外酸性磷酸酶和蛋白酶活性的影响

多数研究表明外生菌根真菌能够促进植物养分吸收并提高植物生长,但是对其发生的原因研究较少。本文在室内控制条件下,研究了真菌菌丝分泌N、P相关胞外酶及其受土壤有机碳（胡敏酸）和无机碳（碳酸钙）添加的影响,结果表明：1）3种真菌——松乳菇(Lactarius deliciosus)、变色红菇（Russula integra）、铆钉菇（Gomphidius viscidus）菌丝均能够分泌酸性磷酸酶和蛋白酶,而且多数情况下,MMN培养基培养14 d时,各个酶活性较高,而不同菌的胞外酶活性存在较大的差异,平均值来看铆钉菇酸性磷酸酶活性最低而蛋白酶活性最高,其它2个真菌菌丝的胞外酶活性差异不大;2）添加胡敏酸后,3种菌丝的酸性磷酸酶活性都是随着胡敏酸添加量的增加而逐渐增加;但蛋白酶活性存在差异：松乳菇的蛋白酶活性随着胡敏酸添加量的增加而逐渐增加;变色红菇的蛋白酶活性对胡敏酸不敏感,受其影响不大;铆钉菇的蛋白酶活力在少量的胡敏酸作用下最强,但浓度过高反而抑制其蛋白酶的活性。3）添加碳酸钙后,总体来看,3种菌丝胞外酸性磷酸酶和蛋白酶活性都是添加少量碳酸钙时酶活性最强,随着浓度的增加(如0.1 g),其酶活性开始受到抑制。综上所述,真菌菌丝能够分泌酸性磷酸酶和蛋白酶,这可能是因为这些外生菌根真菌能够促进植物养分吸收和快速生长的原因;有机碳和无机碳的加入可以直接影响真菌菌丝胞外酶的分泌,进而影响土壤内有机磷和有机氮化合物的分解,显示其在土壤碳循环中的作用。     

小菌虫肠道可培养细菌的分离鉴定 及产消化酶活性分析

摘要：目的 分离小菌虫肠道可培养细菌,并研究其产消化酶活性,探讨肠道细菌对小菌虫消化食物的影响。方法 采用传统细菌分离培养方法分离小菌虫肠道细菌,利用16S rDNA序列进行细菌分子鉴定;利用筛选培养基鉴别各细菌的产蛋白酶、脂肪酶、淀粉酶和纤维素酶活性。结果 在小菌虫肠道中分离到4种可培养细菌,分别是枯草芽胞杆菌（Bacillus subtilis）、肺炎克雷伯菌（Klebsiella pneumoniae）、Pseudocitrobacter faecalis和芽胞杆菌（Bacillus sp.）。其中,2种芽胞杆菌属细菌有产消化酶活性。枯草芽胞杆菌有产纤维素酶、淀粉酶和蛋白酶活性;芽胞杆菌仅有产蛋白酶活性,但产酶能力低于枯草芽胞杆菌。结论 小菌虫肠道细菌中可培养细菌结构简单,但其中的芽胞杆菌属细菌有产纤维素酶、淀粉酶和蛋白酶能力,说明小菌虫肠道中的2种芽胞杆菌属细菌可能有协助小菌虫进行食物消化的功能。     

Characteristics of proteolysis in the gastrointestinal tract in the early postnatal period

The activity of proteinases was increased, when we used experimental casein for artificial feeding of newborn rats. Control casein contained in milk substitutes delayed protein evacuation from the stomach. The activity of intestine pancreatic proteinases was increased when we used experimental casein and decreased, for control casein. When the activity of pancreatic proteinases increased, lysosomal intestine activity decreased.     

人及大鼠胃、小肠CGRP的定位研究

本实验选用成年健康男性尸体和成年雄性Wistar大鼠各4例,死后迅速取胃、小肠各段组织.用免疫组织化学PAP法,对CGRP在人及大鼠胃、小肠中的分布进行了研究.结果表明:胃粘膜内可见CGRP免疫反应阳性的内分泌细胞,其数量人多于大鼠;胃、小肠各段均可见CGRP免疫反应阳性神经纤维分布;大鼠小肠粘膜下层及内环、外纵肌间可见单个存在的CGRP免疫反应阳性神经细胞.这些结果表明,胃和小肠中的CGRP有两种来源,即来源于神经和内分泌细胞.文内还对CGRP在胃和小肠中的可能功能进行了讨论.     

Effects of temperature,pH, and heavy metals (copper,zinc) upon proteinase activities in digestive tract mucosa of typical and facultative piscivorous fish

Species-specific differences and effects of natural (temperature, pH) and anthropogenic (zinc, copper) factors upon activities of casein-and hemoglobinlytic proteinases have been studied in the gut mucosa in pike (Esox lucius), zander (Stizostedion lucioperca), burbot (Lota lota) and perch (Perca fluviatilis). It is revealed that the intestine mucosa proteinase activities differ considerably from those ones in the fish stomach mucosa and that studied factors affect these activities to different extents. The stomach mucosa proteinases are most tolerant to the effects of studied factors, which is especially true for pike. The least tolerant are the intestine caseinlytic proteinases (except for the enzymes in perch). The effects of temperature, pH, and heavy metals depend on fish species, factors’ combinations (mono-factor, bi-factor, or multi-factor action) and on factor variability.     

白芍总苷在正常大鼠体内的组织分布研究

探讨白芍总苷在正常大鼠体内的组织分布特点,为预测其药理作用及不良反应提供依据.正常大鼠按2.82 g/kg灌胃给予TGP药液后1、3、6h取心、肝、脾、肺、肾、胃、小肠、大肠等组织,各组织匀浆后,将匀浆液制成冻干粉,HPLC法测定冻干粉中芍药苷和芍药内酯苷浓度,计算各组织中两者浓度.结果显示1h各组织中均能测到芍药苷和芍药内酯苷,3h除胃和小肠外,其他各组织中两者浓度均达到最大值,小肠、胃、大肠及肾、脾、肝中浓度较高,6h小肠、大肠、胃中浓度较高,其他各组织中浓度较低.说明灌胃TGP后组织分布迅速且广泛,胃、小肠、大肠及肾、脾、肝是主要分布器官,容易在胃肠蓄积,其他组织中蓄积较少,为进一步研究白芍总苷的药理作用及作用机理提供了指导,同时为白芍归经理论提供了一定的现代科学依据.     

Proteinase,Lipase and Amylase Activities in the Small Intestine of Pigs Suffering from Colienterotoxaemia

The activities of proteinases, lipases, amylases and the activities of proteinase inhibitors, as well as the numbers of Escherichia coli in the contents from the small intestine were examined for pigs suffering from colienterotoxaemia and for healthy pigs. Enzyme activities were determined using an agar diffusion test. Compared with healthy animals the activities of proteinases and amylases in diseased animals were reduced while lipases showed increased activity. In pathologically changed contents showing large numbers of E. coli, proteinases could not be demonstrated; however, proteinase inhibitors were found in these contents. In healthy animals, proteinase inhibitors were not demonstrated in ingesta-con-taining contents. In diseased animals, E. coli were found in large numbers in all parts of the small intestine. In healthy animals, E. coli was demonstrated especially in the posterior part of the small intestine and regularly in small numbers. The possible influence of digestive enzymes, especially proteinases and their inhibitors, on enterotoxins from E. coli is discussed.     

兔胃肠道中双歧杆菌的初步研究

双歧杆菌(Bifidobacterium)是兔体及其它某些哺乳动物的重要生理细菌,在微生态学上属于原籍菌群(autochthonous flora)。它是一属革兰氏阳性、无芽胞、无荚膜、无鞭毛、多形性厌氧杆菌。该菌对人、     

Neurotrophin receptors and enteric neuronal development during metamorphosis in the amphibian<Emphasis Type="Italic"> Xenopus laevis</Emphasis>

During metamorphosis, the frog intestine goes through a dramatic shortening with extensive apoptosis and regeneration in the epithelial layer and connective tissue. Our aim was to study changes in the enteric nervous system represented by one inhibitory (vasoactive intestinal polypeptide; VIP) and one excitatory (substance P, neurokinin A; SP/NKA) nerve population and concomitant changes in neurotrophin receptor occurrence during this development in the gut of Xenopus laevis adults and tadpoles at different stages of metamorphosis (NF stages 57–66). Sections were incubated with antibodies against the neurotrophin Trk receptors and p75^NTR, and the neurotransmitters VIP and SP/NKA. Trk-immunoreactive nerves increased dramatically but transiently in number during early metamorphic climax. Nerves immunoreactive for p75^NTR were present throughout the gut, decreased in number in the middle intestine during climax, and increased in the large intestine during late metamorphosis. The percentage of VIP-immunoreactive nerves did not change during metamorphosis. SP/NKA-immunoreactive nerves were first apparent at NF stages 61–62 in the middle intestine and increased in the stomach and large intestine during metamorphosis. Endocrine cells expressing SP/NKA increased in number in stomach, proximal, and middle intestine during metamorphic climax. Thus, neurotrophin receptors are expressed transiently in neurons of the enteric nervous system during metamorphosis in Xenopus laevis and SP/NKA innervation is more abundant in the intestine of the postmetamorphic frog than in the tadpole.This study was supported by grants from the Swedish Research Council to S. Holmgren     

Pinocytosis of Ferritin from the Gut Lumen in Larvae of a Sea Star (Patiria miniata) and a Sea Urchin (Lytechinus pictus)

Pinocytosis of macromolecules from the gut lumen is demonstrated for the first time in larval stages of invertebrates. Developing sea stars ( Patiria miniata ) and sea urchins ( Lytechinus pictus ) were incubated in seawater containing ferritin, which was detected in cell organelles by transmission electron microscopy. Pinocytotic uptake of ferritin by gut cells of Patiria could be detected as soon as the larval mouth opened before the esophagus, stomach and intestine could be distinguished from one another; in contrast, no pinocytosis was detected at the comparable developmental stage (the prism larva) of Lytechinus . Pinocytosis was first detected in developing Lytechinus in pluteus larvae, especially in the stomach and intestine. In gut cells of both kinds of echinoderm larvae, the ferritin progressed rapidly from coated pits at the luminal cell membrane to secondary lysosomes (e.g. this progression took only about 10 min in stomach cells of Patiria larvae). Phagocytosis from the gut lumen was never observed after latex beads or starch granules were fed to larvae of Patiria and Lytechinus . Moreover, there was no evidence of pinocytosis of ferritin or phagocytosis of large particles by epidermal cells of larvae of either species.     

MECHANISM OF ACTIVATION AND POSSIBLE ROLES OF THE CATHEPSIN B-LIKE AND D-LIKE PROTEINASES IN THE EGGS OF PHILOSAMIA CYNTHIA RICINI*

Abstract The mechanism of activation and possible roles in the yolk protein's degradation of the cathepsin B-like and cathepsin D-like proteinases in eggs of Philosamia cynthia ricini were studied. The results showed that acidification could not obviously change the molecular masses of the proteinases. The two proteinases were not likely to modify with each other. The results suggested that the two proteinases might hydrolyze different yolk proteins, or hydrolyze the yolk proteins at different stages during embryogenesis. The results of double immunodiffusion showed that the cathepsin B-like and cathepsin D-like proteinases might exist in the eggs of some different species in Lepidoptera.