Morphological characteristics of cultured olfactory bulb cells

Cultured olfactory bulb cells from embryonic mice had ultrastructural characteristics similar to those of many cell types in the intact adult mouse olfactory bulb. Identified cultured cells included mitral/tufted cells, granule cells, short-axon cells, and fibrous and protoplasmic astrocytes. Cultured neurons were found as individual cells, clusters or aggregates. Clusters consisted of a loose array of neurons that appeared to be densely interconnected by neurites. However, few neurites or fascicles emanated from clusters to adjoining areas. Aggregates consisted of many small, usually rounded, neurons piled on top of one larger neuron, or on more than one, with typically many neurites and fascicles projecting to adjacent aggregates, clusters or individual neurons. Neurites of cultured olfactory bulb cells were well developed, and some were several millimeters long. Synapses were very prominent in these cultures, especially in aggregates, clusters, and fascicles. Electron-lucent, dense-core, and coated vesicles were present. Polarity, shape, and length of the long axis (size) of 815 cultured neurons, identified by positive anti-microtubule-associated protein 2 staining, were documented. Cultured neurons varied in size from 9 to 27 m, with an average size of 16 m. Elliptical bipolar (35%), triangular multipolar (21%), and round unipolar (15%) were the most common polarity/shape combinations found in culture. Multipolar, triangular, triangular multipolar, and elliptical bipolar cells increased in size with increasing age of culture. The relative proportions of triangular, multipolar, elliptical multipolar, and triangular multipolar cells decreased, whereas the relative proportions of round, unipolar, and round unipolar cells increased with increasing age of culture. These changes in population subtypes and cell size may indicate continued differentiation and maturation of cultured neurons.     

Stimulus-dependent activation of c-Fos in neurons and glia in the rat cerebellum

The intent of the present study was to use chemical or electrical stimulation of cerebellar afferents to determine how different stimulation paradigms affect the pattern of activation of different populations of neurons in the cerebellar cortex. Specifically, we analyzed immediate changes in neuronal activity, identified neurons affected by different stimulation paradigms, and determined the time course over which neuronal activity is altered. In the present study, we used either systemic (harmaline) or electrical stimulation of the inferior cerebellar peduncle (10 and 40 Hz) to alter the firing rate of climbing and mossy fiber afferents to the rat cerebellum and an antibody made against the proto-oncogene, c-fos, as a marker to identify activated neurons and glia. In control animals, only a few scattered granule cells express nuclear Fos-like immunoreactivity. Although no other cells show Fos-like immunoreactivity in their nuclei, Purkinje cells express Fos-like immunoreactivity within their somatic and dendritic cytoplasm in control animals. Within 15 min of chemical or electrical stimulation, numerous granule and glial cells express Fos-like immunoreactivity in their nuclei. Cells in the molecular layer express Fos-like immunoreactivity following harmaline stimulation in a time and lobule specific manner; they do not appear to be activated in the electrical stimulation paradigm. Following harmaline injections, there is an initial loss of Fos-like immunoreactivity in the cytoplasm of Purkinje cells; 90 min later, nuclear staining is observed in a few scattered Purkinje cells. Following electrical stimulation, the cytoplasmic staining in Purkinje cells is enhanced; it is never present in the nucleus. Data derived from this study reveal cell-specific temporal and spatial patterns of c-Fos activation that is unique to each paradigm. Further, it reveals the presence of an activity dependent protein in the cytoplasm of Purkinje cell somata and dendrites.     

Effect of Fuzheng Kangbai Granule (扶正抗白冲剂) on Immune Function and Survival Time in Minimal Residual Leukemia Model Mice

Ｍｉｎｉｍａｌｒｅｓｉｄｕａｌｌｅｕｋｅｍｉａ（ＭＲＬ）ｉｓｔｈｅｏｒｉｇｉｎｏｆｌｅｕｋｅｍｉａｒｅｌａｐｓｅ,ｔｈｅｔｒｅａｔｍｅｎｔｏｆＭＲＬｉｓｔｈｅｋｅｙ ｐｒｏｂｌｅｍｉｎ ｐｒｅｖｅｎｔｉｎｇｌｅｕｋｅｍｉａｒｅｌａｐｓｅａｎｄｐｒｏｌｏｎｇｉｎｇｔｈｅｉｌｌｎｅｓｓ ｆｒｅｅｓｕｒｖｉｖａｌｏｆｌｅｕｋｅｍｉａｐａｔｉｅｎｔｓ．ＴＣＭｈｏｌｄｓｔｈａｔｔｈｅｍａｎｉｆｅｓｔａｔｉｏｎｏｆＭＲＬｃｏｎｓｉｓｔｅｄｏｆｔｗｏｃｈａｒａｃｔｅｒｉｓｔｉｃｓ：ｔｈｅ Ｅｖｉｌｅｌｉｍｉｎａ…     

止泻颗粒剂治疗小儿消化不良性肠炎136例

运用止泻颗粒剂与思密达作对照治疗小儿消化不良性肠炎,治疗组136例,对照组80例,2组总有效率分别为95．5％,37．5％,疗效比较具有显著性差异,P＜0．01。     

基于肠道菌群和代谢组学研究化滞柔肝颗粒治疗非酒精性脂肪肝的作用机制

目的 探讨化滞柔肝颗粒治疗非酒精性脂肪肝（nonalcoholic fatty liver disease,NAFLD）的作用机制。方法 高脂饮食饲养8周构建大鼠NAFLD模型,化滞柔肝颗粒ig给药4周。半自动生化分析仪测定大鼠血脂相关指标;油红O染色观察肝脏组织病理变化;Illumina Miseq测序平台对V3～V4可变区进行扩增和测序,超高效液相色谱串联质谱（UPLCMS/MS）开展血浆代谢组学研究,并结合Spearman进行肠道菌群与代谢组学之间的关联分析。结果 与模型组相比,化滞柔肝颗粒中、高剂量组可显著降低大鼠体质量（P<0.05、0.01）和血清总胆固醇（totalcholesterol,TC）、三酰甘油（triacylglycerol,TG）水平（P<0.01）,升高高密度脂蛋白（high-density lipoprotein cholesterol,HDL-C）水平（P<0.05、0.01）。此外,化滞柔肝颗粒高剂量组还可显著降低大鼠低密度脂蛋白（low-density lipoprotein cholesterol,LDL-C）水平（P<0...     

Enhanced neurogenesis after transient global ischemia in the dentate gyrus of the rat

The dentate gyrus is one of the few areas of the mammalian brain where new neurons are continuously produced in adulthood. Certain insults such as epileptic seizures and ischemia are known to enhance the rate of neuronal production. We analyzed this phenomenon using the temporary occlusion of the two carotid arteries combined with arterial hypotension as a method to induce ischemia in rats. We measured the rate of cell production and their state of differentiation with a mitotic indicator, bromodeoxyuridine (BrdU), in combination with the immunohistochemical detection of neuronal markers. One week after the ischemic episode, the cell production in dentate gyrus was increased two- to threefold more than the basal level seen in control animals. Two weeks after ischemia, over 60% of these cells became young neurons as determined by colabeling with BrdU and a cytoplasmic protein (CRMP-4) involved in axonal guidance during development. Five weeks after the ischemia, over 60% of new neurons expressed calbindin, a calcium-binding protein normally expressed in mature granule neurons. In addition to more cells being generated, a greater proportion of all new cells remained in the differentiated but not fully mature state during the 2- to 5-week period after ischemia. The maturation rate of neurons as determined by the calbindin labeling and by the rate of migration from a proliferative zone into the granule cell layer was not changed when examined 5 weeks after ischemia. The results support the hypothesis that survival of dentate gyrus after ischemia is linked with enhanced neurogenesis. Additional physiological stimulation after ischemia may be exploited to stimulate maturation of new neurons and to offer new therapeutic strategies for promoting recovery of neuronal circuitry in the injured brain.     

Serotonergic inhibition of the mossy fibre—granule cell glutamate transmission in rat cerebellar slices

Summary The glutamatergic mossy fibre granule cell pathway has been investigated in rat cerebellar slices. Exposure to 35 mM KCI, a concentration of K⁺ known to elicit Ca²⁺-dependent releases of excitatory amino acids from cerebellar slices, raised cGMP levels. The cGMP response was decreased in a concentration-dependent manner by D-(–)-2-amino-5-phosphonopentanoic acid (D-AP5) and by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) indicating the involvement of ionotropic glutamate receptors of both the N-methyl-D-aspartate (NMDA) and the non-NMDA type. The K⁺-evoked production of cGMP was potently inhibited (EC₅₀ = 1.21 nM) by 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a selective 5-HT₂ receptor agonist. The effect of DOI (0.01 M) was antagonized by 0.03 M of the 5-HT₂ receptor antagonists ketanserin and methiothepin. At concentrations higher than 0.1 M, both antagonists increased on their own the cGMP response elicited by high-K⁺. This effect was insensitive to tetrodotoxin.It had been previously shown that rat mossy fibre endings release glutamate upon depolarization and that such release can be inhibited by activation of 5-HT₂ receptors sited on the mossy fibre endings. Altogether the available data suggest the following conclusions: (a) the glutamate/aspartate endogenously released in cerebellar slices during K⁺ depolarization increase cGMP synthesis through the activation of both NMDA and non-NMDA receptors; (b) a portion of the cGMP response can be prevented by 5-HT₂ receptor activation and may reflect the activity of the mossy fibre-granule cell pathway. Thus serotonin is likely to exert a potent inhibitory control of the excitatory mossy fibre input to the cerebellum by acting at receptors of the 5-HT₂ type. Correspondence to M. Raiteri at the above address     

仁术健胃颗粒逆转胃癌前期病变的临床研究

目的观察仁术健胃颗粒对胃癌前病变的逆转作用。方法将１２０例符合诊断标准的胃癌前期病变患者,随机分为仁术健胃颗粒治疗组８０例和胃复春对照组４０例进行临床疗效观察。结果①综合疗效：治疗组总有效率为９１．２５％,对照组为７５．００％,经统计学处理,差异有显著性（Ｐ〈０．０５）;②治疗组胃镜及对萎缩、肠化、异型增生的疗效分别为７１．２５％、７７．５０％、７９．４９％、８８．４６％,对照组为５２．５０％、６     

肺宝对小鼠免疫功能影响的实验研究

对小白鼠免疫功能影响进行了试验研究 ,试验结果表明 ,肺宝颗粒 1 3.4g/kg(相当生药 2 5 .8g/kg) ,能明显促进鸡红细胞免疫的小白鼠血清溶血素 (IgM)生成 ,具有增强特异性体液免疫功能的作用。 6 .7g/kg(相当生药 1 2 .9g/kg)能促进小白鼠网状内皮系统 (RES)吞噬功能 ,具有增强机体非特异性细胞免疫功能的作用。 1 3.4g/kg对卡介苗 (活菌 )所致小白鼠迟发超敏反应 (Ⅲ型变态反应 ) ,有非常显著的抑制作用 ,能明显提高机体特异性细胞免疫功能。     

高效液相色谱法测定午时茶胶囊中厚朴酚与和厚朴酚含量

目的 :建立中成药午时茶胶囊中厚朴酚与和厚朴酚含量测定的高效液相色谱法。方法 :色谱条件为 AlltimeC1 8(Alltech 5μm,4.6mm× 2 5 mm)柱 ,流动相 :甲醇 -水 (72∶ 2 8) ;流速 :1.0 ml/ min;检测波长 :2 94nm。结果 :测得厚朴酚与和厚朴酚加样回收率为 99.82 % (RSD=0 .89% ,n=6)。结论 :该法可为午时茶胶囊的质量控制和制备工艺提供依据