p16蛋白在甲状腺癌组织表达及临床意义的关系

目的 探讨p16蛋白表达在甲状腺癌发生、发展中的作用及其临床意义。方法采用免疫组织化学SP法检测p16蛋白在甲状腺癌组织中的表达或缺失。结果p16蛋白在甲状腺乳头癌和甲状腺滤泡癌组织中的表达率明显高于甲状腺髓样癌和未分化甲状腺癌组织的表达率(P<0.05)。Ⅰ期p16蛋白的表达率明显高于Ⅲ、Ⅳ期(P<0.05)。无淋巴结转移组p16蛋白的表达率(92.6％)高于有淋巴结转移组(64.7％)(P<0.05)。高分化组p16蛋白的表达率(90.3％)高于低分化组(61.5％)(P<0.05)。结论p16蛋白参与了甲状腺癌的发生和发展,与甲状腺癌的临床分期、分化程度、淋巴结转移有关,可作为临床判定预后的参考指标之一。     

Temporal stability of acetylcholinesterase staining in colonic and rectal neural tissue

Confirmation of the clinical diagnosis of Hirschsprung's disease on standard rectal suction biopsy requires demonstration of aganglionosis in 60 adequate serial sections of submucosa. Positive staining for acetylcholinesterase (AChE), demonstrating an increase in nerve fibres within the lamina propria, muscularis mucosae, and subjacent submucosa, is a useful adjunctive test. In this study, sections of distal colonic muscularis propria and rectal mucosa were stained for AChE over a period of days following storage at 4 °C and at room temperature (15–20 °C). Positive staining of neural tissue was demonstrated in specimens stored at 4 °C for up to 14 days, at which time the experiment was discontinued due to tissue autolysis. Positive staining of the myenteric plexus in colonic specimens stored at room temperature also continued until tissue dissolution became marked at 5 days. This study has demonstrated stability of AChE staining of intestinal neural tissue in specimens stored at 4 °C for 14 days, which suggests that reliable staining for AChE should still be achievable if rectal biopsies are taken in clinics/hospitals without access to staining facilities, provided that tissues are transferred (fresh and moist, at 4 °C) to a reference laboratory for staining within several days of the biopsy procedure. Accepted: 26 November 1996     

False-Positive β-Galactosidase Staining in Osteoclasts by Endogenous Enzyme: Studies in Neonatal and Month-Old Wild-Type Mice

Escherichia coli β-galactosidase (β-gal), encoded by the lacZ gene, has become an essential tool in studies of gene expression and function in higher eukaryotes. lac-Z is widely used as a marker gene to detect expression of transgenes or Cre recombinase driven by tissue-specific promoters. The timing and location of promoter activity is easily visualized in whole embryos or specific tissues using the cleavable, chromogenic substrate, 5-bromo-4-chloro-3-indolyl-D-galactopyranoside (X-gal). The tissue specificity of promoters in transgenic constructs is routinely tested by using a promoter of choice to drive lacZ. Alternatively, the targeted expression of Cre recombinase to perform in vivo recombination of loxP sites can be visualized by β-gal staining in mice carrying a Cre-activated lacZ transgene, such as the ROSA26 strain. In the course of our investigations, we examined β-gal activity in bone tissue from genetically normal mice using standard detection methodology and found very high endogenous activity in bone-resorbing osteoclasts. This was true in frozen, paraffin, and glycol methacrylate sections. X-gal staining colocalized with the osteoclast marker, tartrate-resistant acid phosphatase (TRAP). β-gal activity was present in osteoclasts in long bones, in the mandible, and in both neonatal and more mature animals. We present this brief article as a caution to those testing genetic models of skeletal gene expression using β-gal as a marker gene.     

Dipeptidyl peptidase IV in mammalian lungs

Endothelial cells of the pulmonary circulation are equipped with an ectoenzyme protease system on their luminal surface. The membrane-bound proteases act on the circulating polypeptides and cleave certain peptide bonds in their structure, thus modifying their biological properties. We studied the enzyme dipeptidyl peptidase IV (DP-IV) in mammalian lungs in order to elucidate its contribution to the aforementioned proteolytic processing. We have found that lungs of mammalian species posses DP-IV with different levels of specific activity. In rat lungs the specific activity of DP-IV progressively increased during development between the 18th fetal and the 70th postnatal days. Human embryonal and fetal lungs had significantly higher specific activity of DP-IV compared with the lungs of adult individuals. The enzyme in lungs was mainly membrane bound and was solubilized by some detergents, but not with papain and trypsin. The Triton X-100-solubilized DP-IV from rat lung lysosomal-microsomal membranes migrated during electrophoresis on continuous 4–30% gradient polyacrylamide gel at native apparent M_r values of 260 000 and 490 000. Using a histochemical technique we found the enzyme activity of DP-IV in the capillary bed of the lung alveolar septa only. Four aminoacyl-L-proline-4-nitroanilide substrates for DP-IV were cleaved rapidly during one passage through isolated perfused blood-free rat lungs. The perfusion profiles of cleavage of these substrates were largely coincident with that of Blue Dextran 2 000, a compound, which is unlikely to leave the intravascular space. Taken together, the data suggest that DP-IV operates in vivo as a membrane-bound ectoenzyme on the luminal surface of pulmonary endothelial cells and that it may cleave certain circulating polypeptides.     

Histochemical study of the hepatopancreas in adult females of the pink-shrimp Farfantepenaeus brasiliensis Latreille, 1817

This study provides histochemical data of the hepatopancreatic cells of adult female pink-shrimp (Farfantepenaeus brasiliensis) at two different developmental stages (those with developed gonads and those with exhausted gonads). The F. brasiliensis females were collected in seawater off the Guarapari coast, Espirito Santo, Brazil. Five cell types were identified in this digestive gland: B (vesicular), E (embryonic), F (fibrillar), M (basal) and R (resorptive). The digestive gland was stained with the following techniques: PAS/Alcian blue (for polysaccharides), bromophenol blue (for protein), von Kossa (for bound calcium) and Baker (for lipids). Acid glycoconjugates were found inside vacuoles in the R cells, while neutral polysaccharides were present in the B cells and near to the microvilli. In females with exhausted gonads polysaccharides were also seen in the intertubular spaces and inside the lumina of the tubules. The F and M cells were the most marked by the presence of large amounts of proteins observed in R cells and also inside the vacuoles of B cells. The bound calcium was mainly found in the F and M cells. The F cells showed strong positive staining for lipid while the R cell only stained weakly. The E cells did not react to any of the applied staining techniques. The similarities in the histochemical composition of these hepatopancreatic cells in females with developed gonads, compared to exhausted ones, is justified by the fact that transfer of these elements to the oocytes occurs, in significant quantity, only during the initial stages of gonadal development in F. brasiliensis. Also, they may be more related to the molt stage, as in the case of calcium salts.     

Lyon's Feulgen标准化染色方法与传统Feulgen法的比较研究

目的:Lyon's Feulgen标准法与传统Feulgen法的比较.方法:用相同组织切片经Lyon's Feulgen标准法与传统Feul-gen方法同时染色,观察阳性强度,比较两法的重复性.结果:相同组织切片经Lyon's Feulgen标准法染色阳性产物明显强于传统Feulgen法,重复性更好.结论:Lyon's Feulgen标准法优于传统法.     

Ultrastructural localization of carbonic anhydrase in the vestibular end organs of the guinea pig

Summary Carbonic anhydrase activity was demonstrated cytochemically on an ultrastructural level in the vestibular end organs of the guinea pig. Reaction product was found in the dark cells, transitional cells, cells of the planum semilunatum and supporting cells. In the dark cells, reaction product was observed in the cytoplasm as well as in the basal infoldings. Reaction product was also observed in the basal infoldings of the transitional cells and the cells of the planum semilunatum. The globular structures inside the supporting cells, transitional cells and the cells of the planum semilunatum were also surrounded by the reaction product. These findings suggest that carbonic anhydrase may have different functions, such as water and ion transport, respiration, nutrition and calcium carbonate deposition in the vestibular end organs.     

骨原发性恶性纤维组织细胞瘤的病理形态及免疫组化观察

报告１２例骨原发恶性纤维组织细胞瘤，占本单位同期骨原发肿瘤的１．３％及骨原发恶性肿瘤的４．７％。以四肢长骨多见。临床及Ｘ线皆易误诊为骨肉瘤和骨纤维肉瘤。该瘤由纤维母细胞、组织细胞组成，并有多少不等的多核瘤巨细胞、灶性分布的泡沫细胞及炎细胞。α１－ＡＴ和ＬＹＳＯ是ＭＦＨ的重要标志物，尤α１－ＡＴ敏感，有助于与骨肉瘤及骨纤维肉瘤鉴别，超微结构提示ＭＦＨ来源于原始间叶细胞．     

Basal levels of metabolic activity are elevated in Genetic Absence Epilepsy Rats from Strasbourg (GAERS): measurement of regional activity of cytochrome oxidase and lactate dehydrogenase by histochemistry

The Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are considered an isomorphic, predictive, and homologous model of human generalized absence epilepsy. It is characterized by the expression of spike-and-wave discharges in the thalamus and cortex. In this strain, basal regional rates of cerebral glucose utilization measured by the quantitative autoradiographic [(14)C]2-deoxyglucose technique display a widespread consistent increase compared to a selected strain of genetically nonepileptic rats (NE). In order to verify whether these high rates of glucose metabolism are paralleled by elevated activities of the enzymes of the glycolytic and tricarboxylic acid cycle pathways, we measured by histochemistry the regional activity of the two key enzymes of glucose metabolism, lactate dehydrogenase (LDH) for the anaerobic pathway and cytochrome oxidase (CO) for the aerobic pathway coupled to oxidative phosphorylation. CO and LDH activities were significantly higher in GAERS than in NE rats in 24 and 28 of the 30 brain regions studied, respectively. The differences in CO and LDH activity between both strains were widespread, affected all brain systems studied, and ranged from 12 to 63%. The data of the present study confirm the generalized increase in cerebral glucose metabolism in GAERS, occurring both at the glycolytic and at the oxidative step. However, they still do not allow us to understand why the ubiquitous mutation(s) generates spike-and-wave discharges only in the thalamocortical circuit.     

Immunohistochemical study of distribution of apolipoproteins E and D in human cerebral beta amyloid deposits

Several molecules are known to be closely associated with amyloid deposits in human brain. Among these, apolipoproteins such as apolipoproteins E (apo E) and J (apo J) have been found in two neuropathological hallmarks of Alzheimer's disease (AD) and cerebral amyloid angiopathy (CAA): senile plaques (SPs) and cerebrovascular amyloid. These apolipoproteins may be implicated in amyloid fibrillogenesis. Apo D is a multiligand-multifunctional glycoprotein present in SPs, as we previously reported. The aim of this work is to study the link between immunolocalization of apo E and apo D in AD and CAA brains. Both apolipoproteins were found in all types of SPs, but apo E was observed more often than apo D in mature plaques. Whereas apo E is always located overlapping the amyloid core, apo D seems to situate preferably around and near the amyloid. Immunohistochemistry revealed that these apolipoproteins behave differently in cerebral vessels. Apo E labeling in vessels appears mainly linked to amyloid deposits, whereas apo D shows a distribution almost opposite to that of apo E. This could be an indication of the different roles that each apolipoprotein plays in the pathogenesis of amyloid deposition.