脑损伤后大鼠胰岛B、D细胞和血糖含量的变化

脑损伤后高血糖现象愈来愈受到人们的重视。Sarrat提出海马与胰岛之间存在密切的关系。但脑损伤后大鼠胰岛B、D细胞的变化尚未见报道。本实验通过脑损伤大鼠模型，观察脑损伤后大鼠胰岛B、D细胞的变化，旨在探讨脑损伤对胰岛B、D细胞分泌的影响。     

海洛因依赖对大鼠胰岛A细胞胰高血糖素表达和功能的影响

目的探讨海洛因依赖对大鼠胰岛A细胞胰高血糖素免疫反应（Glu-IR）的影响.方法应用免疫组织化学SABC单染法、形态计量及放射免疫测定法,研究大鼠海洛因依赖期间,A细胞面数密度（NA）及血浆胰高血糖素水平的变化.结果与正常组及盐水组比较,A细胞免疫组织化学显色在海洛因依赖组均增强,阳性细胞增多;A细胞的NA在海洛因依赖第38 d组明显增加,P〈0.01;血浆胰高血糖素水平在海洛因依赖第10 d、24 d、38d组均高于正常组和盐水组,P〈0.01.结论A细胞通过增加细胞数和增强合成分泌胰高血糖素的功能,参与海洛因依赖期间大鼠糖代谢的调节过程.     

白细胞介素I对胰岛B细胞的损伤及其保护

白细胞介素Ｉ对胰岛Ｂ细胞的损伤及其保护方海立，朱文玉（北京医科大学生理教研室北京１０００８３）细胞因子（ｃｙｔｏｋｉｎｅ）是机体在内外环境刺激下产生的一类具有重要生物学功能的细胞调节蛋白。主要包括白介索（ＩＬ－１～１２）、干扰素（ＩＦＮ－α、β、γ）...     

大鼠海洛因依赖期间胰岛B细胞形态学和功能变化的研究

目的探讨胰岛B细胞在海洛因依赖期间的可能作用.方法应用免疫组织化学SABC法、图像分析法和放射免疫测定法,观察大鼠海洛因依赖期间胰岛B细胞免疫组织化学反应、光密度及血清胰岛素水平的变化.结果与正常组和盐水组比较,海洛因依赖组大鼠B细胞免疫反应明显增强,染色加深,且以24d时最为显著.图像分析结果表明,海洛因依赖组B细胞的平均光密度较正常和盐水组显著降低（P＜0.05）,以24 d时间组最低.放射免疫测定结果显示,海洛因依赖组大鼠血清胰岛素（Ins）浓度始终高于正常组和盐水组（P＜0.05）.结论胰岛B细胞的变化表明,在海洛因依赖期间,大鼠胰岛B细胞胰岛素的合成增多,分泌增强,参与了机体在海洛因依赖期间的调节过程.     

人外周血初始B细胞和记忆性B细胞亚群的特征和功能

B淋巴细胞是免疫系统的重要免疫成份,主要功能是介导体液免疫应答。在人外周血中,按照B淋巴细胞的发育阶段及功能的不同,可将B淋巴细胞分为初始成熟B细胞、记忆B细胞和浆细胞。记忆B细胞又可分为IgM记忆B细胞和类型转换的记忆B细胞。近年来的研究表明,B淋巴细胞的亚群远比人想象中的复杂,因此对人B细胞各亚群的起源、发育和功能进行更深入的研究,将有助于治疗自身免疫性疾病,在慢性感染性疾病的治疗过程中找到新的策略,并指导研发安全有效的疫苗。     

葡萄糖、精氨酸影响新生大鼠胰B细胞群的形态定量研究

目的 :研究葡萄糖、精氨酸对新生大鼠胰 B细胞群的影响。方法 :选用 Wistar新生大鼠 ,自出生后第 2日起腹腔注射葡萄糖加精氨酸 ,共 6天。分别于出生后第 10天、2 0天剖腹取胰腺 ,进行常规 HE及免疫组织化学 (PAP)染色 ,显示胰岛及 B细胞。并结合图像分析仪测量胰岛及 B细胞。并结合图像分析仪测量胰岛及 B细胞的直径。结果 :生后 2 0天 ,实验组大鼠胰 B细胞群较第 10天的明显增大 ,而对照组 B细胞群却较第 10天显著缩小 ;第 2 0天两组胰岛直径均较第 10天的小 ,但以对照组的变化明显。结论 :在葡萄糖、精氨酸的作用下 ,新生大鼠胰 B细胞群及胰岛其它内分泌细胞增生     

人外周血初始B细胞和记忆性B细胞亚群的特征和功能

B淋巴细胞是免疫系统的重要免疫成份,主要功能是介导体液免疫应答.在人外周血中,按照B淋巴细胞的发育阶段及功能的不同,可将B淋巴细胞分为初始成熟B细胞、记忆B细胞和浆细胞.记忆B细胞又可分为IgM记忆B细胞和类型转换的记忆B细胞.近年来的研究表明,B淋巴细胞的亚群远比人想象中的复杂,因此对人B细胞各亚群的起源、发育和功能进行更深入的研究,将有助于治疗自身免疫性疾病,在慢性感染性疾病的治疗过程中找到新的策略,并指导研发安全有效的疫苗.     

人外周血初始B细胞和记忆性B细胞亚群的特征和功能

B淋巴细胞是免疫系统的重要免疫成份,主要功能是介导体液免疫应答.在人外周血中,按照B淋巴细胞的发育阶段及功能的不同,可将B淋巴细胞分为初始成熟B细胞、记忆B细胞和浆细胞.记忆B细胞又可分为IgM记忆B细胞和类型转换的记忆B细胞.近年来的研究表明,B淋巴细胞的亚群远比人想象中的复杂,因此对人B细胞各亚群的起源、发育和功能进行更深入的研究,将有助于治疗自身免疫性疾病,在慢性感染性疾病的治疗过程中找到新的策略,并指导研发安全有效的疫苗.     

TLR3激动剂对B细胞功能的影响

目的观察TLR3激动剂Poly I:C对B细胞功能的影响,包括细胞增殖、共刺激分子表达、细胞因子和抗体的分泌情况。方法利用免疫磁珠法分选小鼠脾脏CD19+B细胞,RT-PCR法证实其表达TLR3。体外用Poly I:C刺激B细胞一定时间后,CFSE分裂法检测B细胞增殖,流式细胞术(flow cytometry,FCM)检测B细胞表面共刺激分子表达,CBA(cytometric bead ar-ray)法或ELISA法检测培养上清中细胞因子含量,CBA法检测B细胞分泌Ig的亚型。结果 Poly I:C可以促进B细胞增殖,上调B细胞表面CD40、CD80、CD86和MHC II类分子的表达,促进IL-6和TNF-α的高分泌,诱导IgG1κ抗体的产生。结论 Poly I:C可以通过促进增殖、细胞因子分泌,诱导抗体产生和上调共刺激分子表达等多方面调节B细胞功能。     

斑点免疫金银染色检测RNA抗体及其临床意义

采用斑点免疫金银染色法检测了280份临床血清标本的抗RNA 抗体.其中144份血清同时以ELISA 法作平行对照。两法检测结果差异无显著性(P>0.05),符合率91%,两法定量结果间存在高度的直线正相关性。该抗体在自身免疫病中的检出率为46.6%,其中在SLE 达62%,但在非自身免疫病仅为5.1%,可作为自身免疫病筛查指标,对ANA 阴性者尤有意义。     

Isolation of islets from neonatal pig pancreatic tissue

Summary Pancreatic islet transplantation shows promise as a future method for the treatment of insulin-dependent diabetes mellitus. However, as with reports of whole organ transplants in man, there are likely to be too few human donor organs. One potential approach to try and circumvent this problem is to seek alternative sources of pancreatic tissue. We have studied and developed a technique for isolating functional islets from one-day-old piglets with this in mind. Twelve pancreata, mean weight 1.27±0.36 g, were retrieved aseptically. The islets were isolated using a collagenase digestion procedure. An average of 116,000±45,000 islets/g tissue were isolated from each pancreas. After 24–48 h of culturing, a membrane-boundary was formed around the islets. Dithizone (DTZ) staining revealed greater than 50% of the presumed islets contained at least 5–6 red stained cells (i.e. insulin-containing cells). The insulin content of islet-like structures containing 50%, 10%, and 0% of DTZ stained cells was measured and revealed 32.1, 10.8 and 2.9 µIU, respectively, of immuno-reactive-insulin. In static-perfusion assays, a biphasic insulin secretion was observed in response to increased glucose stimuli. Preliminary studies showed that these islets were able to reverse the hyperglycemic condition of streptozotocin-induced diabetic rats. The present study indicates that one-day-old piglet islets can be utilized as the source of functioning islet cells. Isolation of these cells is reproducible and there appears to be no advantage technically and economically in using fetal pig pancreatic tissue rather than tissue from neonatal piglets for the purpose of obtaining pancreatic islets.     

Preparation and culture of isolated islets and dissociated islet cells from rodent pancreas

Summary Procedures for preparing isolated islets and completely dissociated single islet cells free from the surrounding acinar tissue are described. The mixture of Ficoll and Conray solution (Ficoll-Conray solution) employed for the collection of islets is less viscous and toxic than the plain Ficoll solution and is also applicable for the separation of cells other than islets. Dissociation of islet cells by Dispase results in a satisfactory yield of single islet cells with viability greater than 95%. The islets and dispersed islet cells can be maintained in culture for several months with no deterioration of hormone-producing ability.     

细胞内抗原免疫金银染色技术的改进

为解决细胞内抗原应用免疫金银法染色时背景过重的问题,建立了甘氨酸二次阻断的处理方法,效果较好。     

Effect of a new synthetic serum replacement on insulin and somatostatin secretion from isolated rat pancreatic islets in long-term culture

Summary Serum contains insulin degrading components. We have evaluated the insulin and somatostatin secretion from isolated rat pancreatic islets during a 2-wk culture period using three different serum-containing media, and one serum-free medium with a synthetic serum replacement. Islets incubated in serum-free medium elicited significantly higher daily insulin and somatostatin secretions than islets incubated in the serum-containing media. After a 2-wk culture period, islets from the serum-free medium secreted significantly more insulin and somatostatin than islets cultured in other media when stimulated with 25 mmol/liter glucose together with 15 mmol/liter theophylline. We conclude that the serum-free medium is superior for long-term culture of rat pancreatic islets.     

乙醇对体外培养大鼠大脑皮层神经细胞分化的影响

目的:观察神经细胞微管相关蛋白2(microtubeasso ciated protein-2,MAP2)、孤儿核受体1(nuclear receptor-related factor 1,NURR1)、拓扑异构酶Ⅱβ(DNA topoisomereⅡβ,TopoⅡβ)在乙醇毒性作用下的表达变化,探讨MAP2、NURR1、TopoⅡβ与乙醇导致的神经细胞分化异常的相关关系。方法:取新生鼠(出生24 h内),75%酒精消毒,断头取脑,分离皮层神经细胞接种于培养板,给予剂量75 mmol/L的乙醇培养3 d、5 d、7 d后,采用免疫荧光观察MAP2、NURR1、TopoⅡβ的表达变化;Fluoro-Jade B荧光染色观察神经细胞变性坏死情况。结果:随乙醇染毒时间的延长,原代培养的神经细胞分布较正常对照组稀疏,MAP2的表达发生了变化,以7 d组变化最明显,突起变细变短,阳性表达减弱;NURR1及TopoⅡβ的表达也随乙醇染毒时间的延长,阳性细胞数目及阳性信号表达的强度均呈下降趋势,以7 d最为显著。NURR1阳性细胞计数为(9.6±3.5)与正常对照组的(45.2±3.96)有显著差异(P0.05)。TopoⅡβ(55.8±3.77)与正常对照组的(86.2±4.82)有明显差异(P0.05);Fluoro-Jade B荧光标记显示在乙醇染毒5 d组及7 d组均可见阳性细胞。结论:MAP2、NURR1、TopoⅡβ可能参与了乙醇导致的神经细胞分化异常,改变了细胞骨架稳定,最终导致了部分神经细胞的变性坏死。     

胎鼠胰腺干细胞与胰岛联合移植保护胰岛

背景：胰腺干细胞可在体外维持胰岛的结构,减少胰岛细胞坏死及凋亡,延长胰岛的体外存活时间,保护胰岛的活性。 目的：探索胎鼠胰腺干细胞与胰岛共移植体内保护移植胰岛,提高胰岛移植疗效的可行性。 方法：将成年大鼠35只随机等分为联合移植组、单独胰岛移植组、单纯胰腺干细胞移植组、模型组及对照组,前4组均腹腔注射链脲佐菌素-柠檬酸盐缓冲液建立糖尿病模型。联合移植组、单独胰岛移植组、单纯胰腺干细胞大鼠分别在左侧肾包膜下移植分离纯化孕16 d SD大鼠胎鼠胰腺干细胞和/或成年SD大鼠胰岛。 结果与结论：联合移植组大鼠移植后5 d内血糖可降至正常,血浆胰岛素达到正常水平,胰岛存活时间(18.2±2.4) d;单独移植组大鼠血糖可于移植后1周内降至正常,胰岛存活时间(14.4±2.1) d;两组胰岛存活时间比较差异有显著性意义(P < 0.05)。而其他组糖尿病大鼠血糖均未能降至正常范围。说明胎鼠胰腺干细胞与胰岛共移植可延长胰岛体内存活时间,保护胰岛功能,提高移植疗效。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

蛋白酪氨酸磷酸酶1B的过度表达抑制GFAT的活性

为探讨蛋白酪氨酸磷酸酶1B（PTP1B）在细胞中的过度表达对氨基己糖途径之关键酶谷氨酰胺：6－磷酸果糖酰基转移酶（GFAT）的作用及其可能的作用机制。通过观察PTP1B在细胞内的过度表达对GFAT的活性及其终产物二磷酸尿嘧啶N－乙酰葡萄糖胺生成的影响及PTP抑制剂正钒酸盐的对抗作用，并测定对细胞内GFAT mRNA水平和PTP1B对GFAT的直接作用。发现PTP1B在L6，HepG2,3T3-L1和HIRc等细胞中的过度表达可明显抑制GFAT的活性，且呈剂量和时间相关性。     

Large variability of the intracellular ATP content of human islets isolated from different donors

Observations in experimental heart, liver, kidney and pancreas transplantation indicated that graft function and survival correlates significantly with ATP content of transplanted tissue. The ATP content of cells can be reduced by several factors i.e. the nutritional donor status, storage technique, warm ischemia and cold ischemia time. This study investigates the intracellular ATP content of isolated human islets for the first time. Quantified samples of freshly isolated (digestion-filtration, continuous ficoll gradient purification) and cultured (22°C, CMRL+10% FCS) islet equivalents (IEQ) of consecutively processed human pancreata from multiorgan donors (UW vascular flush) were shock frozen in liquid nitrogen and stored at –196°C until rapid thawing, sonification and subsequent luminometric determination of ATP (Luciferin-Luciferase-reaction) and assessment of islet protein (IP). The ATP content was analysed for freshly isolated and subsequently 5±1 days cultured islets (n=10). The ATP content of freshly isolated human islets was 130.4±53.4 pg/μg IP (mean ± SEM) corresponding to 20.7±6.3 pg/IEQ. After culture ATP content increased to 265.5±113.3 pg/μg IP (204.2±41.5%) corresponding to 43.7±15.3 pg/IEQ (216.1±34.9%; p<0.05). The coefficient of variation was 129.5%, 96.5% (fresh) and 135.0%, 111.0% (cultured) for ATP/μg IP and ATP/IEQ, respectively. The present data show that: (1) the ATP content of freshly isolated human islets varies enormously; (2) intraislet ATP levels increase significantly during 22°C culture suggesting that the capacity to produce ATP is maintained despite hypothermic environment. More data are necessary to clarify the relevance of intraislet ATP content for graft function and survival after islet transplantation.