肾综合征出血热人体组织中病毒包涵体的性质及意义的进一步研究

应用免疫组化、原位分子杂交、电镜及免疫电镜等方法进一步对肾综合征出血热人体尸检组织中病毒包涵体（ＩＢ）的抗原、核酸性质和超微结构特点作了进一步观察。结果,在３９例中的２０例尸检病例组织中显示出病毒核蛋白抗原和血凝素抗抗原阳性的ＩＢ,其中包括１６例陕西尸检病例组织中的６例休克期和１例多尿期病例,１７例上海病例中的３例休克期和９例少尿期病例及３例江西病例中的１例休克期病例。ＩＢ主要分布在呼吸道和肺泡、肾远曲小管和集合管、胃肠道、腺垂体、扁桃体、胰腺、前列腺等组织的粘膜上皮和腺上皮细胞及肝细胞和睾丸生精上皮细胞胞浆中,阳性细胞形态基本正常。应用原位分子杂交,可在该组细胞小同时检测到病毒ＲＮＡ,多为胞浆内弥漫阳性,仅少数组织中显示出病毒ＲＮＡ阳性ＩＢ结构。电镜观察阳性组织细胞中出现由大量微丝微管及颗粒样结构组成的ＩＢ结构,其小的病毒颗粒状结构、内质网及纤维丝状结构呈病毒抗原阳性,上述结构位于高尔基体区。结果说明该病毒有感染上皮细胞的特性,对其宿主细胞的致细胞病变作用是极其温和的,且多表现在亚细胞水平。ＩＢ可能是病毒过量表达抗原的堆积或病毒复制部位,而微丝微管结构可能参与病毒的感染过程。     

慢性缺氧对大鼠肺内皮素表达的影响

本文以ＡＢＣ法和原位杂交技术,观察了慢性缺氧时大鼠肺组织内内皮素－１（ＥＴ－１）的表达情况,结果发现：①正常肺血管内皮细胞有少许ＥＴ－１样阳性染色物质呈现。②缺氧ＩＷ后,肺内ＥＴ－１含量增加,主要位于肺血管内皮细胞和支气管粘膜上皮细胞。③缺氧２Ｗ和３Ｗ后,ＥＴ１阳性免疫物质进一步增加,于肺泡细胞内也见到阳性染色。④缺氧１Ｗ后肺内ＥＴ－１ｍＲＮＡ表达增加,缺氧２Ｗ和３Ｗ后,ＥＴ－１ｍＲＮＡ的表达进一步加强。提示缺氧可刺激肺内ＥＴ－１ｍＲＮＡ的表达,慢性缺氧时肺内ＥＴ－１持续分泌增加,这可能是缺氧性肺动脉高压发生的重要因素之一。     

CO2激光治疗慢性宫颈炎670例疗效观察

本文用ＣＯ２激光对６７０例慢性宫颈炎患者进行了治疗,经复查,治愈率９８％,有效率１００％。     

Platelet activating factor (PAF) stimulates release of PGI2 from inflamed rabbit gallbladder cell cultures

This study examines the hypothesis that PAF stimulates release of PGI₂ from inflamed rabbit gallbladder explant cell cultures. New Zealand white rabbits underwent bile duct ligation for 72 h (72 h BDL), or sham operation, Sham and 72 h BDL gallbladder explants were placed in culture, and the cells grown to 75% confluence. The cells were exposed to increasing concentrations of PAF for 60 min. The media analyzed for eicosanoid release by EIA and the cells analyzed for cyclooxygenase and prostacyclin synthase content by immunoblot analysis. PAF increased release of 6-keto-PGF_1α from the 72 h BDL gallbladder cell cultures in a dose-related manner which was inhibited by indomethacin preincubation by 90%. The increased 72 h BDL cell release of 6-keto-PGF_1α was not associated with changes in the content of cyclooxygenase or prostacyclin synthase. PAF did not alter eicosanoid release from sham control cell cultures. These data suggest that PAF can only up-regulate endogenous 6-keto-PGF_1α release from the 72 h BDL cells that had been previously stimulated by inflammation. PAF may thus contribute to gallbladder distention and injury by chronic stimulation of inflamed gallbladder PGI₂ release.     

Purification and characterization of the high molecular weight microtubule associated proteins from neonatal rat brain

The changes in the levels of microtubule-associated proteins (MAPs) during advanced embryonic stages, neonatal and adult organisms reflect the importance of these cytoskeletal proteins in relation to the morphogenesis of the central nervous system. MAP-1B is found in prenatal brains and it appears to have the highests levels in neonatal rat brains, being a developmentally-regulated protein. In this research, a fast procedure to isolate MAP-1B, as well as MAP-2 and MAP-3 from neonatal rat brains was designed, based on the differential capacity of poly L-aspartic acid to release MAPs during temperature-dependent cycles of microtubule assembly in the absence of taxol. The high molecular weight MAP-1B was recovered in the warm supernatants after microtubular protein polymerization in the presence of low concentrations of polyaspartic acid. Instead, MAP-2 and a 180 kDa protein with characteristics of MAP-3 remained associated to the polymer after the assembly. Further purification of MAP-1B was attained after phosphocellulose chromatography. Isolation of MAP-2 isoforms together with MAP-3 was achieved on the basis of their selective interactions with calmodulin-agarose affinity columns. In addition, MAP-2 and MAP-3 were also purified on the basis of their capacities to interact with the tubulin peptide -II (422–434) derivatized on an Affigel matrix. However, MAP-1B did not interact with the -II tubulin fragment, but it showed interaction with the Affigel-conjugated -I (431–444) tubulin peptide. The different MAPs componentes were characterized by western blots using specific monoclonal antibodies. A salient feature of neonatal rat brain MAP-3 was its interactions with site-directed antibodies that recognize binding epitopes on the repetitive sequences of tau and MAP-2. However, these site-specific antibodies did not interact with MAP-1B from the neonatal rat brain tissue.Abbreviations PAA poly (L-aspartic acid) - HMW-MAPs high molecular weight microtubule associated proteins     

Transient expression of bile-duct-specific cytokeratin in fetal mouse hepatocytes

The differentiation of hepatocytes and biliary epithelial cells has been histochemically analyzed with anti-calf cytokeratin antiserum in the fetal mouse liver. Almost all young fetal hepatocytes transiently express bile-duct-specific cytokeratin; subsequently, the strong staining of the cytokeratin is confined to progenitor cells of intrahepatic biliary epithelial cells around portal veins. These results suggest that all fetal hepatocytes are bi-potent in terms of the differentiation of mature hepatocytes and intrahepatic bile-duct cells, and that the microenvironment around portal veins plays an important role in bile-duct differentiation. Large periportal hepatocytes continue to stain weakly for cytokeratin until 2 weeks after birth, although the number of positive hepatocytes decreases with development. The differentiation of bile ducts from periportal hepatocytes may continue for 2 weeks after birth.     

Rapid regulation of electrolyte absorption in sweat duct

Even though the same Cl channel (CFTR) is common to certain fluid transport functions that are oppositely directed, i.e., secretion and absorption, only fluid secretion has clearly been shown to be acutely regulated. It is now clear that fluid secretion activated by -adrenergic stimulation is controlled by cAMP-mediated opening and closing of CFTR-Cl channels. Since the conductance of the human sweat duct is almost wholly due to CFTR-Cl conductance (CFTR-G_Cl), we sought to determine whether salt absorption via CFTR-Cl channels could also be subject to acute regulation in this purely absorptive epithelium. After -toxin permeabilization, we found that addition of cAMP resulted in a large increase in Cl diffusion potentials across the apical membrane and a more than twofold increase in the average membrane conductance. Since the cAMP effects were dependent on Cl alone, not on Na, and since apical Cl conductance appears to be almost exclusively comprised of CFTR-G_Cl, we surmise that this form of electrolyte absorption like secretion is also subject to acute control through CFTR-G_Cl. Acute regulation of absorption involves both activation by phosphorylation (PKA) and inactivation by dephosphorylation (unknown endogenous phosphatase) of CFTR. Phosphorylation of CFTR was shown by the facts that CFTR-G_Cl could be activated by cAMP and inhibited by the kinase antagonist staurosporine, or by removal of either substrate ATP or Mg²⁺ cofactor. Inactivation of CFTR-G_Cl by endogenous phosphatase(s) was indicated by a spontaneous but reversible loss of CFTR-G_Cl upon removal of cAMP. Such loss of CFTR-G_Cl activity could be prevented either by application of phosphatase inhibitors or by using phosphatase-resistant ATP--S as substrate to phosphorylate CFTR. We surmise that absorptive function is subject to rapid regulation which can be switched on and off acutely by a control system that is common to both absorptive and secretory processes and that this control is crucial to switching between conductive and nonconductive transport mechanisms during salt absorption.The authors are grateful to Mr. Kirk Taylor for expert technical assistance, and to numerous volunteer human subjects for participating in the experiments with informed consent. Supported by grants from the National Institutes of Health, DK-41329-04 and the National Cystic Fibrosis Foundation, Z 439.     

Testicular main ducts and spermatic ducts in some cyprinid fishes I. Morphology, fine structure and histochemistry

Alburnus alburnus, Leuciscus cephalus and Vimba vimba efferent duct systems of the male gonads consist of testicular main ducts and spermatic ducts. These have similar histological, fine structural and (enzyme–) histochemical characteristics and function in (1) storage and (2) nutrition of spermatozoa, (3) synthesis of steroid glucuronides, (4) secretion of proteins and enzymes (5) formation of an ionic gradient in the seminal fluid and (6) they have auto– and heterophagocytotic activities. Therefore testicular main ducts and spermatic ducts are important in the formation of the seminal fluid.     

Extrahepatic tissue copper concentrations in white perch with hepatic copper storage

Hepatic copper storage in man (Wilson's disease), Bedtington and West Highland white terriers, and white perch ( Morone americana ) is characterized by the progressive accumulation of copper in hepatic lysosomes bound to cytoprotective metallothionein. In man, saturation of the liver storage capacity results in the distribution of copper to extrahepatic tissues with multiple organ system dysfunction. To determine if extrahepatic tissue copper concentrations also increase in white perch, copper and zinc levels in liver, brain, heart, gills, serum, and bile were determined by atomic absorption spectrophotometry and compared to striped bass ( Morone saxatilis ). Results showed that brain copper concentrations in. white perch were elevated and significantly correlated with liver copper. Bile and serum copper also increased significantly with liver copper. Copper levels in heart and gill tissues were low. Liver zinc was increased in white perch but not to the same magnitude as copper, and was correlated significantly with liver copper; possibly a non-specific secondary increase related to an overall increase in hepatic metallothionein. Histochemical staining of liver with rubeimc acid for copper was proportional to copper concentrations, and clusters of positive mononuclear cells were also seen in brain and spleen. Foci of macrophages in spleen were also intensely positive with Perl's iron stain which may have been indicative of haemolysis. The patterns of copper distribution seen in white perch present a useful comparative model to study alterations in copper metabolism.     

Chronic Administration of Lithium Chloride Increases Immunodetectable Glial Fibrillary Acidic Protein in the Rat Hippocampus

Abstract: We studied the effect of treating rats with lithium salts on the content and in vitro phosphorylation rate of the astrocyte cell marker, glial fibrillary acidic protein (GFAP), in brain slices. Rats were fed a diet incorporating lithium chloride until the concentration of Li⁺ in serum reached 0.6–1.2 m M , a range similar to that achieved in clinical practice. Hippocampal tissue was analyzed for immunoreactive GFAP by a dot assay, and slices of hippocampus and caudate nucleus were labeled with [³²P]-phosphate to determine the in vitro rate of phosphorylation of GFAP. Compared with controls, the level of immunoreactive GFAP in the hippocampus from lithium-treated rats was increased 34%, and GFAP in hippocampal slices incorporated 39% more ³²P. This effect of lithium was apparently not confined to the hippocampus because the in vitro rate of phosphorylation of GFAP in caudate slices was also increased in the treated rats.