Neurons of the hypothalamus concentrate (3H)progesterone or its metabolites

Selective concentration of [(1)H]progesterone or its metabolites is observed in nuclei of neurons in certain hypothalamic regions of the guinea pig 15 minutes after injection of [1,2,6,7-(3)H]progesterone, by use of dry-mount autoradiography. Highest concentrations of progestin target neurons exist in the nucleus arcuatus, the nucleus preopticus periventricularis, and the nucleus preopticus suprachiasmaticus. Previous administration of unlabeled progesterone inhibits the nuclear concentration of radioactivity, but cortisol has no effect. Estradiol priming enhances the nuclear uptake of radioactivity. The results demonstrate the existence of progestin target sites in the hypothalamus and suggest a direct action of progestin on certain hypothalamic structures.     

( 3 H)lysergic acid diethylamide: cellular autoradiographic localization in rat brain

Intravenous administration of [(3)H]lysergic acid diethylamide(LSD) to rats resulted in accumulation of the drug in the brain within 15 minutes. Autoradiographic methods were used to differentiate free and bound [(3)H]LSD in brain tissue.Free [(3)H]LSD was generally distributed in the pituitary and pineal glands, cerebellum, hippocampus,and choroid plexus.Bound [(3)H]LSD was localized in neurons of the cortex, caudate nucleus, midbrain, and medulla,as well as in choroid plexus epithelium.     

An unusual form of lipid linkage to the CD45 peptide

Some protein kinases and phosphatases are myristoylated on their amino terminus, which perhaps contributes to subcellular localization or regulation. Glycoprotein CD45, a hematopoietic tyrosine phosphatase, was examined for fatty acid content. The CD45 protein incorporated [3H]myristate, but little [3H]palmitate. The label was not metabolized and reincorporated into amino acids or saccharides, as revealed by peptide maps of CD45 labeled with [3H]myristate, 14C-labeled amino acids, [35S]methionine, or 125I, and glycosidase treatments, respectively. The myristate label was resistant to mild alkaline methanolysis and was found in fatty acid and sphingosine, indicating an unusual form of lipid attachment to CD45.     

催产素在广西本地水牛下丘脑-垂体-卵巢轴的免疫组化定位

[目的]研究催产素（OT）在广西本地水牛下丘脑、垂体及卵巢的分布,进而了解OT在下丘脑、垂体及卵巢三者之间分泌释放途径。[方法]采用免疫组织化学SuperPicTureTM二步法检测5头广西本地水牛的下丘脑、垂体和卵巢中催产素（OT）的分布情况。[结果]下丘脑中分泌OT的神经元主要分布在弓状核、视上核及室旁核,在腹内侧核、腹外侧核、交叉上核、背内侧核、乳头体、下丘脑前核等核团也有一定数量的阳性神经元;在腺垂体发现OT免疫反应阳性产物,自垂体柄和正中隆起的一侧可见平行排列的OT阳性神经纤维断续地延伸至神经部;卵巢中只在生殖上皮、卵泡颗粒细胞和黄体细胞发现有大量OT免疫阳性产物。[结论]首次发现OT存在于广西本地水牛下丘脑-垂体-卵巢轴的各个环节,并且下丘脑中各主要核团均有OT免疫阳性神经元的分布,尤其以弓状核、视上核、室旁核分布最多,为进一步研究OT的合成和作用机制提供形态学依据,并对广西本地水牛的繁殖育种及泌乳起到一定的参考指导作用。     

Dopamine: release from the brain in vivo by amantadine

After dopamine stores in the caudate nucleus of cats were labeled with [(3)H]dopamine, the ventricular system was perfused with artificial cerebrospinal fluid. The addition of amantadine to the perfusing fluid caused a doserelated increase in the concentrations of [(3)H]dopamine appearing in the perfusion effluent. Subthreshold concentrations of amantadine also enhanced the efflux of [(3)H]dopamine induced by electrical stimulation of the caudate nucleus.     

Estrogen formation by the isolated perfused rhesus monkey brain

Perfusion of two isolated brains from immature male rhesus monkeys with [(3)H]androstenedione resulted in the identification of free and conjugated [(3)H]estrone and free [(3)H]estradiol from the perfusates. In the dissected cerebral tissues, estrogens were recovered only from the hypothalamus and limbic system. The production of estrogens from androstenedione during the 40-minute perfusions in these two experiments totaled 1.58 and 2.83 nanograms.     

Lipocalin核糖体展示文库的构建及雌二醇特异性模拟抗体的筛选

[目的]构建Lipocalin核糖体展示文库,并筛选雌二醇特异性模拟抗体。[方法]以Lipocalin蛋白家族中的BBP(胆汁三烯结合蛋白)为模板,设计包含16个随机突变位点的引物,合成BBP基因文库;引入核糖体展示所必需的全部组件,采用重叠PCR技术将BBP库与核糖体展示组件进行拼接,构建核糖体展示Anticalin模拟抗体库;再将构建的Anticalin文库进行体外转录与翻译,产生模拟抗体-核糖体-mRNA三联复合体文库。以小分子抗原雌二醇为靶标,采用固相亲和方法筛选抗雌二醇抗体,通过改变Mg2+浓度将模拟抗体-核糖体-mRNA三联复合体解离,得到与模拟抗体对应的mRNA,经过RT-PCR得到筛选后的DNA文库并重新引入核糖体展示元件进行下一轮淘筛。[结果]通过5轮生物淘筛,模拟抗体得到富集,获得高特异性抗雌二醇抗体。其中有一株克隆的亲和力为54 mmol/L(Kon=4.975 6×104个/M·S;Koff=0.002 7个/S)。IC50与LOD值分别为50和0.071 ng/ml。[结论]获得的雌二醇模拟抗体可以作为抗体并用于检测动物体内雌二醇的残留。     

17β-雌二醇在土壤中迁移特征的研究

[目的]为了研究内源甾体雌激素在土壤中的迁移特征。[方法]以17β-雌二醇为对象,采用模拟土柱开展淋滤试验。[结果]表层土壤雌二醇在淋滤过程向深层土壤迁移,土壤有机质对雌二醇的迁移具有迟滞作用;当滤速为O．05L／（s·m2）、雌二醇添加量为1．25mg、淋滤时间为200min时,砂性土壤和黏性土壤中（距顶层土壤1．0m）雌二醇的残留浓度分别为0．06和0．18mg／kg;淋滤液中的腐植酸与土壤中的有机质发生竞争吸附,对E2的迁移起促进作用;而淋滤液中的无机盐（KCI）因促进土壤对E2的吸附,使得雌二醇在土壤中的迁移性能减弱。[结论]该研究可为土壤环境中雌激素污染控制提供科学依据。     

Pituitary gonadotrophs: nuclear concentration of radioactivity after injection of ( 3 H)testosterone

Gonadotrophs and castration cells in the male rat pituitary showed nuclear concentration of radioactivity 1 hour after [1,2,6,7-(3)H]testosterone injection. Thyrotrophs and acidophils did not retain radioactivity; also the cells of the intermediate and posterior lobes did not accumulate radioactivity. The autoradiographic results suggest a direct and selective action of androgen on gonadotrophs, which contrasts with the action of estradiol which was shown earlier to bind not only to basophils but to acidophils and chromophobes as well.     

陶粒负载生物滴滤塔净化恶臭气体研究(英文)

[目的]探讨净化单一恶臭气体H2S和NH_3工艺条件及影响因素。[方法]以陶粒作为生物滴滤塔的填料负载微生物,考察了生物滴滤塔于挂膜启动及稳定运行阶段对H_2S和NH_3的降解性能。[结果](1)净化H_2S废气的生物滴滤塔系统在9 d内即完成挂膜,在进口浓度100-1 000 mg/m~3,空床停留时间(EBRT)142-290 s的条件下,H_2S的去除率均可达到99%以上;(2)净化NH_3废气的生物滴滤塔在10 d内即完成挂膜,对NH_3去除率均达到94.61%。[结论]适宜条件下,生物滴滤塔对于H_2S和NH_3的去除率均较高。     

Sequence and expression of human estrogen receptor complementary DNA

The mechanism by which the estrogen receptor and other steroid hormone receptors regulate gene expression in eukaryotic cells is not well understood. In this study, a complementary DNA clone containing the entire translated portion of the messenger RNA for the estrogen receptor from MCF-7 human breast cancer cells was sequenced and then expressed in Chinese hamster ovary (CHO-K1) cells to give a functional protein. An open reading frame of 1785 nucleotides in the complementary DNA corresponded to a polypeptide of 595 amino acids and a molecular weight of 66,200, which is in good agreement with published molecular weight values of 65,000 to 70,000 for the estrogen receptor. Homogenates of transformed Chinese hamster ovary cells containing a protein that bound [3H]estradiol and sedimented as a 4S complex in salt-containing sucrose gradients and as an 8 to 9S complex in the absence of salt. Interaction of this receptor-[3H]estradiol complex with a monoclonal antibody that is specific for primate ER confirms the identity of the expressed complementary DNA as human estrogen receptor. Amino acid sequence comparisons revealed significant regional homology among the human estrogen receptor, the human glucocorticoid receptor, and the putative v-erbA oncogene product. This suggests that steroid receptor genes and the avian erythroblastosis viral oncogene are derived from a common primordial gene. The homologous region, which is rich in cysteine, lysine, and arginine, may represent the DNA-binding domain of these proteins.     

东北小鲵端脑发生和发育的电生理学特性研究

[目的]探讨东北小鲵（ Hynobius leechii）端脑发生与发育的电生理学特性。[方法]应用微电极电生理技术,对东北小鲵端脑发生和发育不同时期的自发电活动进行在体胞外记录。[结果]东北小鲵端脑发生和发育各时期自发放电形式共有单个放电、连续单个放电、簇状放电、连续簇状放电和不规则放电五种形式。端脑的发生阶段为3种,以单个放电和连续单个放电为主,记录到少数簇状放电;端脑的发育阶段为5种,以单个放电为主。随着端脑发生和发育的进行,胞外放电幅度逐渐提高,放电持续时间增加,放电频率加快,并于成体期达到最大;单个放电时程逐渐缩短;其它放电形式伴随发生发育过程相继出现。[结论]随着东北小鲵端脑发生与发育,神经元的兴奋性逐步提高,神经元电活动形式逐渐多样化。     

Posttranslational glutamylation of alpha-tubulin

The high degree of tubulin heterogeneity in neurons is controlled mainly at the posttranslational level. Several variants of alpha-tubulin can be posttranslationally labeled after incubation of cells with [3H]acetate or [3H]glutamate. Peptides carrying the radioactive moiety were purified by high-performance liquid chromatography. Amino acid analysis, Edman degradation sequencing, and mass spectrometric analysis of these peptides led to the characterization of a posttranslational modification consisting of the successive addition of glutamyl units on the gamma-carboxyl group of a glutamate residue (Glu445). This modification, localized within a region of alpha-tubulin that is important in the interactions of tubulin with microtubule-associated proteins and calcium, could play a role in regulating microtubule dynamics.     

Effects of Shenpang acupoint-stimulation on estrogen receptor immunoreactive neurons in thalamus of rabbits

To investigate the effects of Shenpang acupoint-stimulation in reproductive endocrinology, the changes in estrogen receptor immunoreactive (ER-IR) neurons after Shenpang acupoint-stimulation were studied by using immunohistochemistry. ER-IR positive reactions were detected in most nuclei of the thalamus. In the acupuncture-treated group, a great number of ER-IR positive neurons with clear dendrites existed in the nucleus, paraventricular nucleus, ventrolateral nucleus, ventromedial nucleus, ventroprincipal nucleus, centromedian nucleus, reticular nucleus, and periventricular nucleus of thalamus, and they were mainly located in the cytoplasm, nucleus and neutrite, and some also existed in the cytoplasmic membrane. In contrast, a few neurons existed in the above-mentioned nuclei in the control group, but they were slightly stained. It is concluded that Shenpang acupoint-stimulation can promote the expression of estrogen receptors in the above nuclei. Translated from Acta Veterinaria et Zootechnica Sinica, 2006, 37(1): 71–75 [译自: 畜牧兽医学报]     

GnRH在青年摩杂一代水牛下丘脑-垂体-卵巢轴的免疫组化定位

[目的]研究促性腺激素释放激素（GnRH）在青年摩杂一代水牛下丘脑-垂体-卵巢轴的免疫组化定位。[方法]采用免疫组织化学SuperPicTure^TM二步法检测5头青年摩杂一代水牛下丘脑、垂体和卵巢中GnRH的分布情况,研究GnRH在下丘脑、垂体及卵巢的分布上的联系,进而了解GnRH在下丘脑、垂体及卵巢三者之间分泌释放的途径。[结果]GnRH存在于青年摩杂一代水牛下丘脑-垂体-卵巢轴的各个环节。青年摩杂一代水牛下丘脑中各主要核团都有GnRH免疫阳性神经元的分布,尤其以视上核、视前核、室旁核分布较多。对于垂体,GnRH免疫阳性产物只存在于青年摩杂一代水牛的腺垂体中。青年摩杂一代水牛卵巢中生殖上皮细胞、颗粒细胞和黄体细胞发现有GnRH免疫阳性产物。[结论]为进一步研究GnRH在生殖中的生理和作用机制提供形态学依据,并对青年摩杂一代水牛的繁殖育种起到参考指导作用。     

Transfer of a protein encoded by a single nucleus to nearby nuclei in multinucleated myotubes

Specialized regions of muscle fibers may result from differential gene expression within a single fiber. In order to investigate the range of action of individual nuclei in multinucleated myotubes, C2 myoblasts were transfected to obtain stable cell lines that express a reporter protein that is targeted to the nucleus. Hybrid myotubes were then formed containing one or a few transfected nuclei as well as a large number of nuclei from the parental strain. In order to determine how far the products of a single nucleus extend, transfected nuclei were labeled with [3H]thymidine before fusion and the myotubes were stained to identify the reporter protein. In such myotubes the fusion protein was not confined to its nucleus of origin, but was restricted to nearby nuclei.     

禽流感病毒H1、H3亚型和M基因三重PCR检测方法的建立

【目的】建立用于检测禽流感病毒（Avianinfluenzavirus,AIV）且能同时区分H1和H3亚型AIV的方法,为有效防控禽流感（Avianinfluenza,AI）奠定基础。【方法】根据H1亚型、H3亚型AIV-HA基因和M基因的保守核苷酸序列,分别设计3对特异性引物,以含有H1亚型AIV和H3亚型AIV的cDNA为模板,对三重PCR扩增条件进行优化,验证其特异性和敏感性,并对临床样品进行AIV检测。【结果】H1和H3亚型AIV混合模板经三重PCR扩增可获得3条特异性条带,其中302bp为H1亚型HA基因、453bp为M基因、626bp为H3亚型HA基因;含有H1或H3亚型AIV的模板均出现两条特异性条带,大小分别为302和453bp、453和626bp;含有其他亚型AIV的模板仅出现一条特异性条带（453bp）;而其他禽呼吸道疾病病毒均未扩增出任何条带。优化后的三重PCR最低能同时检测出7.00pg的H1亚型AIV、3.62pg的M基因和20.60pg的H3亚型AIV;对100个临床样品进行AIV检测,AIV总阳性率为36%,其中H1亚型阳性率4%、H3亚型阳性率9%、其他亚型阳性率23%。【结论】禽流感病毒H1、H3亚型和M基因三重PCR检测方法具有快速、敏感、特异等特点,适用于AIV的检测及H1和H3亚型AIV的分型检测。     

Insulin rapidly increases diacylglycerol by activating de novo phosphatidic acid synthesis

The mechanisms whereby insulin increases diacylglycerol in BC3H-1 myocytes were examined. When [3H]arachidonate labeling of phospholipids was used as an indicator of phospholipase C activation, transient increases in [3H]diacylglycerol were observed between 0.5 and 10 minutes after the onset of insulin treatment. With [3H]glycerol labeling as an indicator of de novo phospholipid synthesis, [3H]diacylglycerol was increased maximally at 1 minute and remained elevated for 20 minutes. [3H]Glycerol-labeled diacylglycerol was largely derived directly from phosphatidic acid. Insulin increased de novo phosphatidic acid synthesis within 5 to 10 seconds; within 1 minute, this synthesis was 60 times greater than that of controls. Thus, the initial increase in diacylglycerol is due to both increased hydrolysis of phospholipids and a burst of de novo phosphatidic acid synthesis. After 5 to 10 minutes, de novo phosphatidic acid synthesis continues as a major source of diacylglycerol. Both phospholipid effects of insulin seem important for generating diacylglycerol and other phospholipid-derived intracellular signaling substances.     

Dopaminergic terminals in the rat cortex

The destruction of ascending noradreniergic pathways by bilateral microinjections of 6-hydroxydopamnine made laterally to the pedunculus cerebellaris superior completely abolished the in vitro synthesis of [(3)H]norepinephrine from L-[(3)H]tyrosine in slices and in synaptosomes of the rat cortex. However, normal [(3)H]dopamine synthesis could still be observed in both cortical preparations from animals with lesions. These results provide the first biochemical support for the existence of dopaminergic terminals independent of noradrenergic terminals in the rat cortex.     

蔗糖和锌对水稻根系生长的调节(英文)

[目的]研究锌和蔗糖与水稻根系生长发育的关系。[方法]以水稻中花11号为试验材料,分析了不同浓度的锌、蔗糖、甘露醇、锌+蔗糖和锌+甘露醇处理条件下水稻根系生长、活性氧产生和根系质子分泌的变化。[结果]在1~3mmol/L范围内随着锌浓度的增加,水稻根系总长度、数量及活性氧的产生均增加,但质子分泌水平降低。除了株高外,初生根长、初生根侧根长/数、不定根长/数和不定根侧根长/数都不同程度的受到蔗糖浓度的影响,高浓度蔗糖诱导活性氧增加,无糖或高浓度蔗糖都导致根系质子分泌能力降低。但是,在相同浓度的蔗糖处理条件下锌诱导这些指标的变化与无锌处理的明显不同。5%蔗糖和5%甘露醇对根系生长的调节有显著差异,说明该调节作用是由蔗糖信号而非渗透势引起的。[结论]该研究表明蔗糖和锌对水稻根系的生长都具有重要的调节作用。