The USDA trypsin inhibitor study. I. Background,objectives, and procedural details

Short-term feeding studies have shown that raw soy flour and purified trypsin inhibitors (TI) cause pancreatic hypertrophy and hyperplasia in certain monogastric animals. Prolonged exposure to high levels of TI in raw soy ultimately leads to pancreatic nodular hyperplasia and acinar cell adenoma in rats exposed to low levels of pancreatic carcinogens. Such change has been observed even in the absence of any exposure to known carcinogens. These results emphasized the need for chronic (2 yr) feeding trials which would clearly reveal dose response relationships of alterations in pancreatic pathology to dietary TI and possible interactions with dietary protein and fat.Here we report on the objectives and designs of the first phase of the USDA TI Study, including composition of diets, preparation, and analyses of test substances. To provide the requisite TI and protein levels, raw and heated defatted soy flours, soy protein isolates, and lactic casein were used. The interrelationship of dietary level of TI (93–1270 mg/100 g diet) and that of protein (10%, 20%, and 30%) to pancreatic function and pathology will be reported in the following three papers. Both serial and chronic sacrifice regimes, respectively, were employed with 26 diets and 40 weaning male Wistar rats per dietary group.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

仔猪胰腺细胞体外培养体系的建立

为了研究体外培养仔猪胰腺细胞的适宜条件,试验选取1日龄的仔猪胰腺组织置于冰上清洗、修剪,对消化酶、离心方式、培养皿铺被方式、培养液及添加不同促生长因子等多种因素进行筛选和优化。结果表明:0.25%胰酶消化所获得的细胞强于0.1%胶原酶消化获得的细胞,细胞清亮,易于生长;采用500 r/min离心5 min、多次离心所获得的细胞较Percoll不连续密度梯度离心和5%BSA等密度梯度离心所获得的细胞贴壁率高,生长能力强,易于培养传代;采用0.1%明胶或20μg/mL的层黏连蛋白铺被培养皿均比无铺被的培养皿获得更多的贴壁细胞;以RPMI 1640、M199、F12、L-DMEM多种培养液培养仔猪胰腺细胞,表现出多种细胞形态和生长趋势;在基础培养液中添加上皮生长因子、角质化生长因子、β-巯基乙醇和白血病细胞抑制因子、转铁蛋白与亚硒酸盐混合试剂等促干细胞增殖因子可促进不同形态细胞的增殖。说明按此方式培养可获得大量、多种细胞形态的仔猪胰腺祖/干细胞。     

Suspected congenital hyperinsulinism in a Shiba Inu dog

A 3-month-old male intact Shiba Inu dog was evaluated for a seizure disorder initially deemed idiopathic in origin. Seizure frequency remained unchanged despite therapeutic serum phenobarbital concentration and use of levetiracetam. The dog was documented to be markedly hypoglycemic during a seizure episode on reevaluation at 6 months of age. Serum insulin concentrations during hypoglycemia were 41 U/μL (reference range, 10-29 U/μL). The dog was transitioned to 4 times per day feeding, diazoxide was started at 3.5 mg/kg PO q8h, and antiepileptic drugs were discontinued. No clinically relevant abnormalities were identified on bicavitary arterial and venous phase contrast computed tomographic imaging. The dog remained seizure-free and clinically normal at 3 years of age while receiving 5.5 mg/kg diazoxide PO q12h and twice daily feeding. Seizures later occurred approximately twice per year and after exertion, with or without vomiting of a diazoxide dose. Blood glucose curves and interstitial glucose monitoring were used to titrate diazoxide dose and dosing interval. Congenital hyperinsulinism is well recognized in people but has not been reported in veterinary medicine.     

鸡胚胎胸腺、胰腺、性腺的组织发生

在孵化的第4d,鸡胚胸腺的原基发生于第Ⅲ、Ⅵ咽囊的背壁上皮。到第9d,胸腺被间充质分叶。胸腺小体发生于第13d。在第15d,胸腺的皮质与髓质开始分化。到20～21d,在封闭的微血管周围,由数层网状细胞形成典型的胸腺小体。背侧胰芽及腹侧胰芽发生于孵化的第3～4d。到第15d,外分泌部由初期的复管状腺变成复管泡状腺。胰岛细胞团在第11d发生。随着鸡胚的发育,胰岛的数目逐渐增多。生殖嵴在第3d发生。到第4d,从卵黄囊内胚层来的原性细胞,便向雌性左侧的性腺转移。初级性索仲入髓质形成间质细胞。次级性索形成卵泡细胞。第5d时,雄性的初级性索,仲入间充质内形成曲细精管及间质细胞群的原基。第8d以后,性腺的性别可以区分。     

Isolation and properties of lipolysis inhibitory proteins from wheat germ and wheat bran

Proteins inhibiting pancreatic lipasein vitro have been isolated from wheat germ and wheat bran, with relative molecular mass ranging from 24,400 to 27,500. Inhibition of pancreatic lipase by the wheat germ proteins is related to their ability to interact with the emulsified substrate and to hinder the adsorption of the enzyme on the interface. The extent of inhibition depends on the amount of substrate and is independent of the enzyme concentration. Bile salts forming micelles in the concentration range used are able to partially reverse the inhibition of pancreatic lipase by the wheat germ proteins. The nutritional significance of the data obtained is discussed.     

MCT1抑制剂p-CMB对乙酰胆碱诱导的大鼠胰液分泌的影响

采用在体试验方法研究单羧酸转运蛋白第1亚型（monocarboxylate transporter 1,MCT1）抑制剂（p-chloromercuribenzene sulfonate,p-CMB）对乙酰胆碱（acetylcholine,Ach）诱导的大鼠胰液分泌的影响。Spra-gue-Dwley（SD）雄性大鼠,体重180g-210g,分6组,每组5只,试验前禁食24h,自由饮水。动物麻醉后经外科手术后,每隔10min收集一次胰液,采用Lowry法测定胰液蛋白量,Bernfeld法测定胰酶。结果对照组（-p-CMB）和实验组（＋p-CMB）,在注射1、5、10μg／kg的Ach后对胰液分泌量、胰蛋白分泌量以及胰淀粉酶分泌量都呈现增加作用,注射后1h内达到峰值,之后逐渐降低。然而,试验组的胰液分泌各项指标比对照组的显著减少（P〈0．01）。表明MCT1抑制剂p-CMB减弱了Ach增加大鼠胰液分泌的作用。     

亚砷酸对人胰腺癌PANC-1细胞增殖及RASSFIA基因甲基化的影响

目的探讨亚砷酸对人胰腺癌PANC-1细胞增殖及RAS相关结构域家族基因1A（RASSHA）甲基化的影响。方法PANC-1细胞用不同浓度亚砷酸处理,采用MTT法检测细胞增殖,甲基化特异性PCR（MSV）法检测RASSFlA基因甲基化情况。结果随着亚砷酸浓度增加（1．2～4．8μmol／L）和作用时间延长,PANC-1细胞抑制率逐渐增加（P〈0．05）。PANC－1细胞中RASSFlA基因表现为高甲基化状态,但其甲基化随亚砷酸浓度增加而逐渐减弱。结论亚砷酸对PANC－1细胞增殖的抑制作用具有时间和剂量依赖性,并通过RASSFlA基因的去甲基化起作用。     

Pancreatic duct obstruction in the pig: electron microscopy of chronic pancreatitis

Ultrastructural alterations of pancreatic tissues in growing pigs caused by surgical ligation of the main pancreatic duct are described. Five animals aged 6 to 7 weeks were operated on and necropsied 2, 6 or 8 weeks later. In addition, 3 pigs that were sham-operated served as controls. Obstruction of the ducts caused progressive luminal dilatations and rupture of intercellular connections of the epithelium followed by destruction and atrophy of acinar parenchyma and proliferations of immature ductular epithelial cells free from zymogen granules.