r－干扰素对喉癌细胞系HEP－2HLA－DR抗原和增殖细胞核抗原表达的影响

本实验用人重组ｒ－干扰素（ｒｈｕ－ＩＦＮ）作用ＨＥＰ－２细胞后ＨＬＡ－ＤＲ抗原和增殖细胞核抗原（ＰＣＮＡ）表达的检测来探讨ｒ－干扰素对ＨＥＰ－２细胞ＨＬＡ－ＤＲ抗原表达诱导作用及体外抗增殖活性。用单克隆抗体ＣＲ３／４３（抗ＨＬＡ－ＤＲ）和Ｋｉ－６７（抗ＰＣＮＡ）。以链霉素一生物素技术（ＬＳＡＢ）检测ＨＥＰ－２细胞ＨＬＡ－ＤＲ抗原和ＰＣＮＡ表达,结果显示：ｒ－ＩＥＮ诱导ＨＬＡ－ＤＲ抗原和抑制ＰＣＮＡ表达其强弱与ｒ－ＩＦＮ剂量有关。资料提示：ｒ－ＩＦＮ不仅对ＨＥＰ－２细胞有细胞毒作用,同时能调节其细胞膜特性,因而在喉癌的治疗中是有效的。     

新疆阿尔泰山驼鹿的初步考察

新疆阿尔泰山驼鹿的初步考察ＡＰＲＥＬＩＭＩＮＡＲＹＩＮＳＰＥＣＴＩＯＮＯＮＴＨＥＥＬＫ（ＡＬＣＥＳＡＬＣＥＳ）ＩＮＸＩＮＪＩＡＮＧＡＬＴＡＩＭＯＵＮＴＡＩＮＫｅｙｗｏｒｄｓＸｉｎｊｉａｎｇ;Ｅｌｋ（Ａｌｃｅｓａｌｃｅｓ）１９９４年７－８月,我们受新疆...     

半衰期延长获得PAI－1抗性的t—PA突变体的构建、表达及特性分析

用基因重级及定位突变技术成功地构建了ｔ－ＰＡ的Ｋ１区缺失突变体ｔ－ＰＡｄｅｌＫ１、ＰＡＩ－１结合位点缺失突变体ｔ－ＰＡｄｅｌ（２９６－３０２）及两的组合突变全ｔ－ＰＡｄｅｌ（Ｋ１,２９６－３０２）,并在ＣＯＳ－７细胞中实现三的暂时性表达,在ＣＨＯ细胞中实现了ｔ－ＰＡｄｅｌ（Ｋ１,２９６－３０２）的稳定性表达。对表达产物的生物学特性分析表明,ｔ－ＰＡｄｅｌ（２９６－３０２）及ｔ－ＰＡｄｅｌ（Ｋ１     

老年大鼠单核／巨噬细胞分泌和表达肿瘤坏死因子增多

为研究老年时肿瘤坏死因子（ＴＮＦ－α）分泌及表达的改变,在内毒素（ＬＰＳ）１．０μｇ／ｍｌ刺激大鼠单核／巨噬细胞后,用酶联免疫法（ＥＬＩＳＡ）测定培养液中ＴＮＦ－α含量,用半定量逆转录－聚合酶链反应（ＲＴ－ＰＣＫ）测定ＴＮＦ－αｍＲＮＡ。同时测定培养液中一氧化氮（ＮＯ）和前列腺素Ｉ２（ＰＧＩ２）的含量。结果显示：老年鼠ＴＮＦ－α分泌量及其ｍＲ－ＮＡ明显高于青年鼠。ＮＯ产量在老年鼠与青年鼠之间无明显差异。老年鼠ＰＧＩ２分泌明显低于青年鼠。由于ＰＧ能抑制ＴＮＦ－α释放,从而推测,ＰＧＩ２产生能力的降低可能是老年大鼠单核／巨噬细胞ＴＮＦ－α分泌量明显高于青年鼠的原因之一。     

人胎气管内胰岛淀粉样多肽免疫反应细胞的个体发生(英文)

为了进一步探讨胰岛淀粉样多肽（ＩＡＰＰ）的分布、定位以及它与其他生物活性物质的关系;用ＩＡＰＰ组织化学ＰＡＰ邻片双标法,观察了１８例１４～３８周人胎气管内ＩＡＰＰ免疫反应（ＩＲ）细胞的个体发生及与５羟色胺（５－ＨＴ）的关系。结果显示,胎１４周,气管粘膜表面的假复层柱状上皮中已有ＩＡＰＰ－ＩＲ细胞（Ｆｉｇ．１＆２）;１５周开始,粘膜固有层气管腺导管上皮中也出现分散的ＩＡＰＰ－ＩＲ细胞（Ｆｉｇ．３）;随胎龄增长,１７～２１周,气管上皮内ＩＡＰＰ－ＩＲ细胞逐渐增多;免疫染色加深（Ｆｉｇ．４＆５）,有些细胞发出细突直达腔面（Ｆｉｇ．６＆７）,粘膜下层的气管腺腺泡中也有ＩＡＰＰ－ＩＲ（Ｆｉｇ．８）; ２２～３８周,气管内 ＩＡＰＰ－ＩＲ 细胞又呈逐渐减少趋势,ＩＡＰＰ－ＩＲ仅出现在基底锥形细胞中（Ｆｉｇ．９＆１０）,且免疫染色较深。邻片未显５－ＨＴ－ＩＲ。本研究表明,人胎儿期气管上皮细胞内有ＩＡＰＰ的表达;且ＩＡＰＰ－ＩＲ细胞随胎期的发育而发生变化。     

人类嗜T细胞白血病Ⅰ型病毒TAX蛋白对NF—kB转录因子的作用

人类嗜Ｔ细胞白血病Ⅰ型病毒（ＨＴＬＶ－Ⅰ）是成人Ｔ细胞白血病（ＡＴＬ）的致病因子,其编码的ＴＡＸ蛋白的反式激活在白血病形成中有重要作用。ＮＦ－ｋＢ是细胞活化和产生细胞因子的重要转录调控因子。正常情况下,ＮＦ－ｋＢ因子与抑制性蛋白ＩＫＢ结合,形成复合物存在于胞质中。ＴＡＸ蛋白可与ＩＫＢ激酶γ（ＩＫＫγ）直接结合,而后启动ＴＡＸ对ＩＫＫα和ＩＫＫβ的结合,并使之发生磷酸化。后者使ＩＫＢ蛋白降解,ＮＦ－     

抗心律失常药UK－68798对家兔心脏房室结区不同细胞的电生理作用

在离体家兔ＡＶＮ区标本上,用微电极技术研究了Ⅲ类抗心律失常新药ＵＫ－６８７９８对ＡＮ,Ｎ,ＮＨ,Ｈ４种细胞的电生理效应。浓度５×１０－９至５×１０－６ｍｏｌ／Ｌ的ＵＫ－６８７９８对４种细胞的动作电位幅值（ＡＰＡ）、静息膜电位（ＲＰ）皆无影响。对ＡＶＮ的自搏率有剂量依赖性减慢作用,但不改变Ａ－Ｈ传导时间。在５×１０－８－５×１０－６ｍｏｌ／Ｌ剂量范围,此药使动作电位时程（ＡＰＤ５０）和（ＡＰＤ９０）发生剂量依赖性延长。４种细胞中以Ｎ细胞的ＡＰＤ５０和ＡＰＤ９０延长百分率最高。各种细胞ＡＰＤ９０延长百分率的排列次序为Ｎ＜ＡＮ＜Ｈ＜ＮＨ,当浓度为５×１０－６ｍｏｌ／Ｌ时的延长百分率分别为９５±２６％（Ｎ）,７５±２２％（ＡＮ）,６３±２６％（Ｈ）,４６±２６％（ＮＨ）。在ＵＫ－６８７９８的作用下,４种细胞的有效不应期（ＥＲＰ）也发生剂量依赖性延长,但不存在像ＡＰＤ延长百分率那样的差别。此外,４种细胞ＥＲＰ所相当的复极化膜电位未受药物影响,从而避免了由于兴奋性恢复的不均一性,使ＡＶＮ区成为折返性心律失常的发源地．     

恒河猴肺内CGRP及CGRP mRNA免疫组织化学和原位杂交的研究

本实验采用免疫组织化学ＡＢＣ－ＧＤＮ法和地高辛标记寡核苷酸探针原位杂交技术,研究了降钙素基因相关肽（ＣＧＲＰ）及其ｍＲＮＡ在恒河猴内的表达。结果显示ＣＧＲＰ分布于恒河猴肺内各级支气管粘膜上皮的神经内分泌细胞（ＮＥＣ）和神经上皮样小体（ＮＥＢ）中。ＣＧＲＰ杂交阳性细胞的分布与免疫组织化学的结果相同。ＣＧＲＰ ｍＲＮＡ杂交信号均匀分布于整个细胞质,而ＣＧＲＰ免疫反应物仅在神经内分泌细胞的基部更明显,说     

新麦草的组织培养及染色体分析

新麦草的组织培养及染色体分析孙震晓,夏光敏,陈惠民（山东大学生物系．济南２５０１００）ＴＩＳＳＵＥＣＵＬＴＵＲＥＡＮＤＣＨＲＯＭＯＭＥＡＮＡＬＹＳＩＳＯＦＰｓＡＴＨＹＲＯＳＡＣＨＹＳＪＵＮＣＥＡ（ＦＩＳＣＨ．）ＮＥＶＳＫＩｓｕｎＺｈｅｎ－ｘｉａｏ;ｘ...     

用蛋白质工程方法改变葡萄糖异构酶最适pH和最适温度

用寡核苷酸诱导的定点突变方法构建了葡萄糖异构酶基因的突变体（Ｎ１８４Ｄ和Ａ１９８Ｃ）。含突变体的重组质粒ｐＴＫＤ－ＧＩ１（Ｎ１８４Ｄ）和ｐＴＫＤ－ＧＩ２（Ａ１９８ｃ）在Ｅ．ｃｏｌｉＫ３８菌株中表达,用ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ ＦＦ和Ｓｅｐｈａｃｒｙｌ Ｓ－３００ＨＲ柱层析分离纯化突变酶。与野生型葡萄糖异构酶比较实验表明：（１）突变酶Ｎ１８４Ｄ的最适ｐＨ值下降了１个单位;等电点下降了０．６个单位     

豚鼠小肠神经节丛的NADPH—黄递酶组织化学观察

目前已知,ＮＡＤＰＨ－－黄递酶组化法可选择性地显示－－氧化氮合成酶（ＮＯｓｙｎｔｈａｓｅ,ＮＯＳ）神经元。因此,我们以ＮＡＤＰＨ－黄递酶组化法,观察了豚鼠小肠肌间神经丛和粘膜下神经丛的神经网格以及ＮＯＳ神经元。结果表明,三段小肠肌间神经丛的神经网眼大小和形态有明显差异,与对应的粘膜下神经丛相比,差异更显著。在肌间神经丛中,ＮＡＤＰＨ－黄递酶阳性神经元胞体大小不等;其长突起伸入节间束,而短突起较多,并可见短突起彼此连接．构成节内偶见的局部神经元回路。从小肠上段到下段,ＮＯＳ神经元数量呈下降趋势。在粘膜下神经丛,我们也观察到少数ＮＯＳ神经元。     

Neuronal distribution and subcellular localization of HCNP-like immunoreactivity in rat small intestine

A novel peptide, hippocampal cholinergic neurostimulating peptide (HCNP), originally purified from young rat hippocampus, affects the development of specific cholinergic neurons of the central nervous system in vitro. In this study, HCNP-like-immunoreactive nerve processes and nerve cell bodies were identified by electron microscopic immunocytochemistry in the rat small intestine. Labeled nerve processes were numerous in the circular muscle layer and around the submucosal blood vessels. In the submucosal and myenteric plexuses, some HCNP-like-immunopositive nerve cell bodies and nerve fibers were present. The reaction product was deposited on the membranes of various subcellular organelles, including the rough endoplasmic reticulum, Golgi saccules, ovoid electron-lucent synaptic vesicles in axon terminals associated with submucosal and myenteric plexuses, and the outer membranes of a few mitochondria. The synaptic vesicles of HCNP-like-positive terminals were 60–85 nm in diameter. The present data provide direct immunocytochemical evidence that HCNP-like-positive nerve cell bodies and nerve fibers are present in the submucosal and myenteric plexuses of the rat small intestine. An immunohistochemical light microscopic study using mirror-image sections revealed that in both the submucosal and myenteric ganglia, almost all choline acetyltransferase (ChAT)-immunoreactive neurons were also immunoreactive for HCNP. These observations suggest (i) that HCNP proper and/or HCNP precursor protein is a membrane-associated protein with a widespread subcellular distribution, (ii) that HCNP precursor protein may be biosynthesized within neurons localized in the rat enteric nervous system, and (iii) that HCNP proper and/or HCNP precursor protein are probably stored in axon terminals.     

Development of the submucous plexus in the large intestine of the mouse

In the small intestine of both embryonic birds and mammals, neuron precursors aggregrate first at the site of the myenteric plexus, and the submucous plexus develops later. However, in the large intestine of birds, the submucosal region is colonised by neural-crest-derived cells before the myenteric region (Burns and Le Douarin, Development 125:4335-4347, 1998). Using antisera that recognize undifferentiated neural-crest-derived cells (p75NTR) and differentiated neurons (PGP9.5), we examined the colonisation of the murine large intestine by neural-crest-derived cells and the development of the myenteric and submucosal plexuses. At E12.5, when the neural crest cells were migrating through and colonising the hindgut, the hindgut mesenchyme was largely undifferentiated, and a circular muscle layer could not be discerned. Neural-crest-derived cells migrated through, and settled in, the outer half of the mesenchyme. By E14.5, neural-crest-derived cells had colonised the entire hindgut; at this stage the circular muscle layer had started to differentiate. From E14.5 to E16.5, p75NTR- and PGP9.5-positive cells were observed on the serosal side of the circular muscle, in the myenteric region, but not in the submucosal region. Scattered, single neurons were first observed in the submucosal region around E18.5, and groups of neurons forming ganglia were not observed until after birth. The development of the enteric plexuses in the murine large intestine therefore differs from that in the avian large intestine.     

Distribution and localization of neurokinin A-like immunoreactivity and neurokinin B-like immunoreactivity in rat peripheral tissue

Using specific radioimmunoassays and immunocytochemistry for neurokinin A (NKA) and neurokinin B (NKB), distribution and localization of these peptides in rat peripheral tissues were studied. NKA-like immunoreactivity (NKA-LI) was present in highest levels of 15.7–23.9 pmol/g wet wt. and NKB-like immunoreactivity (NKB-LI) was in levels of 0.33–0.67 pmol/g wet wt., throughout the gastrointestinal tract involving stomach, duodenum, jejunum, ileum and colon. Immunocytochemical analysis of gastrointestinal tract revealed that NKA-LI and NKB-LI localized in ganglia of both the submucosal and myenteric plexuses as well as varicose neurons in the mucosa and the muscle layer of the small and large intestine. On the other hand, high levels of NKB-LI were observed in oesophagus (0.83 ± 0.08 pmol/g wet wt.), adrenal (1.02 ± 0.21), head of pancreas (0.73 ± 0.06) and kidney (0.98 ± 0.05).

The present study shows the difference of localization of NKA-LI and NKB-LI in peripheral tissues and suggests that NKB may have some physiological role differing from that of NKA in peripheral tissues.     

Immunohistochemical analysis of substance P-containing neurons in rat small intestine

The neuropeptide substance P (SP) is involved in the regulation of epithelial secretion and motility in the rat small intestine. The morphology, chemical profiles and proportion of SP-containing enteric neurons in this tissue have been examined by immunohistochemical analysis of whole-mount preparations obtained from colchicine-treated rats. In the submucosal plexus of the duodenum, jejunum and ileum, the proportion of SP-positive neurons is 53%, 51% and 49%, respectively. All SP-positive submucosal neurons are positive for neurofilament 200 (NF-200) and calretinin. Immunoreactivity for calcitonin gene-related peptide (CGRP) is detectable in 55% of the SP-positive submucosal neurons. Some SP-positive submucosal neurons have two or more long processes emerging from an oval or round cell body, a characteristic of the Dogiel type II neuron (type II neuron; a putative intrinsic primary afferent neuron). About one-third of the neurons in the myenteric plexus are positive for SP and a majority of them are NF-200/calretinin-positive type II neurons. Immunoreactivity for the SP receptor neurokinin-1 receptor (NK1R) has been detected mainly in the submucosal and myenteric NF-200-positive neurons, which are expected to contain SP. These neurons possibly stimulate each other via SP release. Most of the submucosal and myenteric neurons, including type II neurons, show immunoreactive for the prostaglandin E2 receptor EP3 receptor (EP3R). Thus, SP/NF-200/calretinin/NK1R/EP3R is the common chemical profile of type II neurons in the rat small intestine. The proportion of SP-immunopositive submucosal neurons (49%–53%) is higher in the rat small intestine than in the colon (≤11%) and around 50% are positive for CGRP.     

Transient expression of the calcitonin receptor by enteric neurons of the embryonic and early post-natal mouse

Calcitonin receptor-immunoreactivity (CTR-ir) was found in enteric neurons of the mouse gastrointestinal tract from embryonic day 13.5 (E13.5) to post-natal day 28 (P28). CTR-ir occurred in cell bodies in ganglia of the myenteric plexus extending from the esophagus to the colon and in nerve cells of the submucosal ganglia of the small and large intestines. CTR-ir was also found in vagal nerve trunks and mesenteric nerves. Counts in the ileal myenteric plexus revealed CTR-ir in 80% of neurons. CTR-ir was clearly evident in the cell bodies of enteric neurons by E15.5. The immunoreactivity reached maximum intensity between P1.5 and P12 but was weaker at P18 and barely detectable at P28. The receptor was detected in nerve processes in the intestine for only a brief period around E17.5, when it was present in one to two axonal processes per villus in the small intestine. In late gestation and soon after birth, CTR-ir was also evident in the mucosal epithelium. The perinatal expression of CTR within the ENS suggests that the calcitonin/CTR system may have a role in the maturation of enteric neurons. Signals may reach enteric neurons in milk, which contains high levels of calcitonin.     

Divergent fate and origin of neurosphere-like bodies from different layers of the gut

Enteric neural stem cells (ENSCs) are a population of neural crest-derived multipotent stem cells present in postnatal gut that may play an important role in regeneration of the enteric nervous system. In most studies, these cells have been isolated from the layer of the gut containing the myenteric plexus. However, a recent report demonstrated that neurosphere-like bodies (NLBs) containing ENSCs could be isolated from mucosal biopsy specimens from children, suggesting that ENSCs are present in multiple layers of the gut. The aim of our study was to assess whether NLBs isolated from layers of gut containing either myenteric or submucosal plexus are equivalent. We divided the mouse small intestine into two layers, one containing myenteric plexus and the other submucosal plexus, and assessed for NLB formation. Differences in NLB density, proliferation, apoptosis, neural crest origin, and phenotype were investigated. NLBs isolated from the myenteric plexus layer were present at a higher density and demonstrated greater proliferation, lower apoptosis, and higher expression of nestin, p75, Sox10, and Ret than those from submucosal plexus. Additionally, they contained a higher percentage of neural crest-derived cells (99.4 ± 1.5 vs. 0.7 ± 1.19% of Wnt1-cre:tdTomato cells; P < 0.0001) and produced more neurons and glial cells than those from submucosal plexus. NLBs from the submucosal plexus layer expressed higher CD34 and produced more smooth muscle-like cells. NLBs from the myenteric plexus layer contain more neural crest-derived ENSCs while those from submucosal plexus appear more heterogeneous, likely containing a population of mesenchymal stem cells.     

What neurons hide behind calretinin immunoreactivity in the human gut?

Calretinin (CALR) is often used as an immunohistochemical marker for the histopathological diagnosis of human intestinal neuropathies. However, little is known about its distribution pattern with respect to specific human enteric neuron types. Prior studies revealed CALR in both myenteric and submucosal neurons, most of which colabel with choline acetyl transferase (ChAT). Here, we specified the chemical code of CALR-positive neurons in small and large intestinal wholemounts in a series of 28 patients. Besides other markers, we evaluated the labeling pattern of CALR in combination with vasoactive intestinal peptide (VIP). In colonic submucosa, CALR and VIP were almost completely colocalized in about three-quarters of all submucosal neurons. In the small intestinal submucosa, both the colocalization rate of CALR and VIP as well as the proportion of these neurons were lower (about one-third). In the myenteric plexus of both small intestine and colon, CALR amounted to 11 and 10 %, respectively, whereas VIP to 5 and 4 % of the whole neuron population, respectively. Colocalization of both markers was found in only 2 and 3 % of myenteric neurons, respectively. In section specimens, nerve fibers coreactive for CALR and VIP were found in the mucosa but not in the muscle coat. Summarizing the present and earlier results, CALR was found in at least one submucosal and two myenteric neuron populations. Submucosal CALR+/VIP+/ChAT± neurons innervate mucosal structures. Furthermore, CALR immunoreactivity in the myenteric plexus was observed in morphological type II (supposed primary afferent) and spiny type I (supposed inter- or motor-) neurons.     

Computer-aided morphometric analysis of the developing concentric structure of the human fetal intestinal tube

The Image-Pro Plus 3.0 morphometric program was used to study the region-specific organization of the human fetal intestine across the radial axis of the gut at weeks 12 and 18 of gestation. The thicknesses of the epithelium, the submucosa, the muscular layers and the myenteric ganglia were measured in resin-embedded semithin sections. Statistical analysis of the collected data was performed by using the two-way ANOVA, the SNK test and the Pearson correlation. The structural changes relating to the gut morphogenesis within this developmental period were followed both light and electron microscopically. The various tissues forming the radial axis of the intestinal tube exhibited different trends concerning their individual development. The thickness of the epithelium did not change in the fetal period investigated, although the epithelial surface displayed characteristic ultrastructural changes. The thickness of the submucosal layer increased significantly, but with different dynamics along the longitudinal axis, whereas the increases in size of the muscular layers and the myenteric ganglia did not differ significantly along the longitudinal axis of the embryonic intestine. The Pearson correlation revealed a significant correlation between the development of the circular muscle layer and that of the myenteric plexus along the whole length of the intestinal tube. The epithelium, the submucosa and the longitudinal muscle layers developed independently between weeks 12 and 18 of gestation.