虎斑颈槽蛇胸腺APUD细胞的免疫组织化学观察

虎斑颈槽蛇胸腺ＡＰＵＤ细胞的免疫组织化学观察ＴＨＥＩＭＭＵＮＯＨＩＳＴＯＣＨＥＭＩＣＡＬＯＢＳＥＲＶＡＴＩＯＮＯＦＴＨＥＴＨＹＭＩＣＡＰＵＤＣＥＬＬＳＩＮＴＨＥＳＮＡＫＥＲＨＡＢＤＯＰＨＩＳＴＩＧＲＩＮＡＫｅｙｗｏｒｄｓＲｈａｄｂｄｏｐｈｉｓｔｉｇｒ...     

乌鳢胃肠内分泌细胞的免疫组织化学研究

乌鳢胃肠内分泌细胞的免疫组织化学研究夏立群,王庆堂（同济医科大学组织胚胎学教研室武汉４３００３０）关键词乌鳢,内分泌细胞,免疫组织化学ＰＡＰ方法ＩＭＭＵＮＯＣＹＴＯＣＨＥＭＩＣＡＬＳＴＵＤＹＯＦＴＨＥＥＮＤＯＣＲＩＮＥＣＥＬＬＳＩＮＴＨＥＧＡＳＴＲＯ...     

低氧对高原鼠兔和大白鼠肝脏重要金属元素的影响

低氧对高原鼠兔和大白鼠肝脏重要金属元素的影响ＴＨＥＥＦＦＥＣＴＳＯＦＨＹＰＯＸＩＡＯＮＩＭＰＯＲＴＡＮＴＭＥＴＡＬＥＬＥＭＥＮＴＳＩＮＴＨＥＬＩＶＥＲＯＦＰＬＡＴＥＡＵＰＩＫＡＡＮＤＲＡＴ微量元素在动物生长、发育、繁衍等生命过程中具有重要的生理生化...     

青心酮对妊娠高血压综合征胎盘血管内皮和平滑肌细胞内一氧化氮合酶的影响

本文对正常孕妇、妊娠高血压综合征（ＰＩＨ）患者和经青心酮（ＤＨＡＰ）治疗的ＰＩＨ患者等共２４例,应用组织化学分析方法观察胎盘血管内皮细胞（ＶＥＣ）和平滑肌细胞（ＶＳＭＣ）内一氧化氮合酶（ＮＯＳ）活性的变化。结果表明：正常孕妇胎盘ＶＥＣ和ＶＳＭＣ内ＮＯＳ活性较高;ＰＩＨ胎盘ＶＥＣ和ＶＳＭＣ内ＮＯＳ活性明显减弱,并伴有组织和细胞的形态学损伤;经ＤＨＡＰ治疗后的ＰＩＨ胎盘ＶＥＣ和ＶＳＭＣ细胞ＮＯＳ活性较未经ＤＨＡＰ治疗者明显增加,其组织和细胞损伤也减轻。本研究结果提示胎盘内ＶＥＣ和ＶＳＭＣ细胞的ＮＯＳ减少可能与ＰＩＨ的发生和／或发展有关,青心酮治疗ＰＩＨ的作用可能与ＤＨＡＰ促进胎盘ＶＥＣ和ＶＳＭＣ内一氧化氮（ＮＯ）合成有关。     

梨、李两种铁甲虫

梨、李两种铁甲虫ＴＷＯＳＰＥＣＩＥＳＯＦＤＡＣＴＹＬＩＳＰＡＥＸＩＣＩＳＡＡＮＤＰＬＡＴＹＰＲＩＡＳＰ．ＯＦＰＥＡＲＡＮＤＰＬＵＭＱｉＳｈｉｃｈｅｎｇＨｕａｎｇＢａｎｇｋａｎＬｉｕＳｉＪｕｎＸｉｅＹｉｄｉ（ＦｕｊｉａｎＡｇｒｉｃｕｌｌｕｒａｌｕｎｉｖ...     

PIC-BE诱导K562/ADM细胞凋亡及逆转其MDR的研究

β榄香烯吗素（ＰＩＣ－ＢＥ）是抗癌新药β榄香烯的水溶性衍生物．采用人红白血病的多药耐药性（ＭＤＲ）细胞株Ｋ５６２／ＡＤＭ作为实验模型,观察ＰＩＣ－ＢＥ对Ｋ５６２／ＡＤＭ细胞的生长抑制和凋亡诱导作用,并进而研究其对该细胞ＭＤＲ的可能影响．结果显示：（１）Ｋ５６２／ＡＤＭ细胞对ＡＤＭ具有明显的抗性,与Ｋ５６２细胞相比,抗性倍数约为４０倍,而两者对ＰＩＣ－ＢＥ的ＩＣ５０接近,无显著差异;（２）ＰＩＣ－ＢＥ（１０．０～３０．０μｇ／ｍｌ）对Ｋ５６２／ＡＤＭ细胞具有明显的生长抑制和凋亡诱导作用,两种作用的强度在一定的范围内均具药物浓度和作用时间依赖性;（３）低毒剂量ＰＩＣ－ＢＥ（１０．０μｇ／ｍｌ）与ＡＤＭ（４．０μｇ／ｍｌ）联合应用,可显著增强ＡＤＭ对该细胞的生长抑制和凋亡诱导作用,升高细胞内ＡＤＭ的浓度,降低该细胞对ＡＤＭ的ＩＣ５０,使该细胞对ＡＤＭ的抗性有数倍逆转．上述结果提示,ＰＩＣ－ＢＥ不仅是一种有效的广谱抗肿瘤剂,而且也是一种有效的ＭＤＲ逆转剂     

地塞米松和肺成纤维细胞对大鼠胎肺表面活性物质合成的影响

为提高ＤＥＸ对新生儿呼吸窘迫综合征（ＮＲＤＳ）的防治,本文研究了地塞米松（ＤＥＸ）促进肺表面活性物质（ＰＳ）合成的作用机制。用分离培养的２０天大鼠胚胎肺泡Ⅱ型细胞为材料,观察了单独用ＤＥＸ、用与不用ＤＥＸ刺激后的成纤维细胞条件培养液（ＤＥＸＦＣＭ、ＦＣＭ）对肺泡Ⅱ型细胞ＰＳ中磷脂合成及特异性肺表面活性物质蛋白质ＳＰＢ、ＳＰＣｍＲＮＡ表达的影响。结果表明,ＤＥＸ本身及未用ＤＥＸ刺激的ＦＣＭ对肺泡Ⅱ型细胞的上述三种指标均没有影响,而ＤＥＸ刺激后的ＤＥＸＦＣＭ使这三个指标有不同程度的增加效应。提示糖皮质激素刺激肺泡Ⅱ型细胞ＰＳ合成增加是由成纤维细胞介导的     

东北鼢鼠种群生长指标的主分量分析

东北鼢鼠种群生长指标的主分量分析ＰＲＩＮＣＩＰＡＬＣＯＭＰＯＮＥＮＴＡＮＡＬＹＳＩＳＯＦＧＲＯＷＴＨＩＮＤＥＸＯＦＰＯＰＵＬＡＴＩＯＮＦＯＲＭＡＮＣＨＵＲＩＡＮＺＯＫＯＲ（ＭＹＯＰＡＬＡＸＰＳＩＬＵＲＡＳ）东北鼢鼠（Ｍｙｏｐａｌａｘｐｓｉｌｕｒａｓ...     

重组含有Epstein—Barr病毒潜伏膜蛋白1（LMP1）基因的杆状病毒在昆虫？…

用杆状病毒表达系统重组病毒,在昆虫细胞中表达了完整的含有ＥＢＶ－ＬＭＰ１基因３个外显子开放读码框架的长２．３ｋｂ的ｃＤＮＡ片段。用重组病毒感染Ｓｆ９细胞,用免疫荧光染色,结果表明：４８小时表达重组蛋白,７２小时细胞较完整,免疫荧光染色强阳性,９６小时后细胞出现破碎。我们采集７２小时的组织培养上清和细胞破碎裂解液,分别采用ＳＤＳ－ＰＡＧＡＥ、ＨＰＬＣ分子筛法,用免疫蛋白印迹法实验证明,表达的蛋白能被     

河南曹岗湖浮游动物野外现场试验初报

河南曹岗湖浮游动物野外现场试验初报蔡庆华,伍焯田,黎道丰,梁彦龄（中国科学院水生生物研究所,武汉４３００７２）关键词浮游动物,野外现场试验,湖泊生态系统,黄淮海平原ＰＲＩＬＩＭＩＮＡＲＹＲＥＰＯＲＴＯＮＩＮＳＩＴＵＥＸＰＥＲＩＭＥＮＴＯＦＺＯＯＰＬＡ...     

Identification of cultured cell employing specific substance--with special references of "reserve cells" in uterine cervix

Identification of endocervical "reserve cell", which have been regarded as the origin of squamous cell carcinoma of the uterine cervix, was attempted employing immunohistochemically specific substances. The antigenicity of keratin, squamous cell carcinoma antigen(SCC), epithelial membrane antigen(EMA), tissue polypeptide antigen(TPA), vimentin, secretory component(SC) and placental alkaline phosphatase(PLAP) was investigated in histological preparations as well as cultured cells obtained from primary culture of endocervical tissue. The immunohistochemical findings in histological preparations revealed the following: a strongly positive reaction with TPA, a slightly positive reaction with EMA, a very slightly positive with SCC and PLAP, and a negative reaction with keratin, vimentin and SC. Cultured cells were divided into 4 groups according to their morphological characteristics; among these, small rounded or polygonal cells with a centric single nucleus showed similar immunocytochemical reactions to those of "reserve cells" in histological preparations, indicating that "reserve cell" can be growing in culture. The results obtained suggest that immunohistocytochemical specific substances may be useful to identify cultured cells.     

Immunohistochemical diagnosis of preinvasive germ cell cancer of the testis

The aim of the study was to identify testicular carcinoma in situ (CIS), a precursor of germ cell tumours (GCT), in patients from the high risk groups, using classic and alternative immunohistochemical methods. 70 patients with 46,XY karyotype were examined. Whole gonads or biopsy specimens were fixed in Bouin's fluid. In cases with dysgenetic male pseudohermaphroditism (DMP), gross histopathology revealed sex cord tumour gonadoblastoma in 4 and malignant dysgerminoma in 1 out of 23 patients. In all patients, paraffin sections were treated with antibodies against placental-like alkaline phosphatase (PLAP), a classic immunohistochemical marker of GCT and CIS. CIS was detected immunohistochemically in 10 out of 23 cases with DMP (43.5%), in 1 out of 10 cases with androgen insensitivity syndrome (10%), in 3 out of 18 cases operated previously because of already developed GCT in contralateral testis (16.6%) and in 1 out of 3 patients with cryptorchidism in anamnesis (33.3%). CIS was not found in 16 examined adult infertile men with azoospermia. In addition to PLAP investigation, 12 cases with DMP and 6 cases with GCT were examined using M2A and TRA-1-60 antibodies, the alternative immunohistochemical markers of CIS. While in DMP positive reactions for M2A and TRA-1-60 accompanied PLAP reaction in 1/3 of cases, M2A accompanied PLAP in all cases with GCT. The positive reaction for TRA-1-60 accompanied PLAP and M2A in 1 case with GCT. The results indicate that among different risk groups the highest incidence of CIS occurs in DMP. Screening for CIS is of importance also in cryptorchidism, androgen insensitivity syndrome and in men who underwent gonadectomy because of unilateral GCT. The immunostaining for PLAP seems to be more discriminative procedure. The positive staining of CIS cells with M2A and TRA-1-60 antibodies may be indicative for more advanced neoplastic transformation.     

Mutation in aging mice occurs in diverse cell types that proliferate postmutation

To determine the relationship between aging, cell proliferation and mutation in different cell types, hearts, brains and kidneys from G11 PLAP mice between 1 week and 24 months of age were examined. Mutant cells were detected in tissue sections by staining for Placental Alkaline Phosphatase (PLAP) activity, an activity that marks cells that have sustained a frameshift mutation in a mononucleotide tract inserted into the coding region of the human gene encoding PLAP. The number of PLAP(+) cells increased with age in all three tissues. The types of cells exhibiting a mutant phenotype included cells that are proliferative, such as kidney epithelial cells, and cells that do not frequently replicate, such as cardiac muscle cells and neurons. In the brain, PLAP(+) cells appeared in various locations and occurred at similar frequencies in different regions. Within the cerebellum, PLAP(+) Purkinje cell neurons appeared at a rate similar to that seen in the brain as a whole. PLAP(+) cells were observed in kidney-specific cell types such as those in glomeruli and collecting tubules, as well as in connective tissue and blood vessels. In the heart, PLAP(+) cells appeared to be cardiac muscle cells. Regardless of tissue and cell type, PLAP(+) cells occurred as singletons and in clusters, both of which increased in frequency with age. These data show that age-associated accumulation of mutant cells occurs in diverse cell types and is due to both new mutation and proliferation of mutant cells, even in cell types that tend to not proliferate.     

Interactions of lectins with specific cell types in toad urinary bladder. Surface distribution revealed by colloidal gold probes and label fracture

Colloidal gold probes were used in conjunction with pre-embedding labeling and label-fracture to show the plasma membrane distribution of Helix pomatia lectin (HPL) and wheat germ lectin (WGL) binding sites on different epithelial cell types of toad urinary bladder. Mitochondria-rich cells were virtually unlabeled with HPL, but showed a strong affinity for WGL. Granular cells were weakly labeled with WGL but had a variable affinity for HPL. Strongly labeled granular cells were arranged in either chains or clusters that were surrounded by poorly-stained granular cells. By label-fracture, the distribution of gold-labeled lectins was related to other membrane features seen in freeze-fracture. Neither HPL nor WGL binding sites appeared to be specifically related to the large intramembrane particles that characterize granular cells, or to the rod-shaped intramembrane particles that are a feature of membranes of mitochondria-rich cells. The preferential lectin binding affinity of these functionally distinct cell types provides an important starting point for their isolation and the characterization of their plasma membranes. Furthermore, the label-fracture approach can now be used to examine the plasma membrane modifications that occur in these cells under different physiologic conditions affecting epithelial transport processes.     

Clustered distribution of human placental alkaline phosphatase on the surface of both placental and cancer cells. Electron microscopic observations using gold-labeled antibodies

The cell-surface distribution of human placental alkaline phosphatase (PLAP) on cultured cancer cells, A431 and HeLa TCRC-1, and on normal syncytial cells of placental tissue was examined in immunoelectron transmission microscopy using the gold-labeling technique. Chemically fixed cells were reacted with affinity-purified rabbit polyclonal antibodies to PLAP, and the antibodies were visualized using gold particles tagged with goat antirabbit IgG. On all cells PLAP was observed in clusters distributed throughout the membrane surface, including microvilli, but it was not expressed in desmosomes or along other dense regions on the membrane. Previous histochemical and immunochemical techniques failed to demonstrate clusters. The results show that (1) the gold-labeling technique allows a more precise localization of PLAP on the cell surface than previously employed methods, and (2) the distribution of the enzyme is the same on cultured cancer cells and on normal placental syncytial cells. The clustered distribution of PLAP is thus a general phenomenon and is probably influenced by the physiological function of the enzyme, which has yet to be defined.     

Nuclear DNA content and proliferative potential of human carcinoma in situ cells in testes of intersex children

Gonocytes, fetal germ cells, when persisted beyond infantile period of life are considered as preinvasive testicular germ cell cancer (carcinoma in situ--CIS). The aim of the study was to investigate nuclear DNA content and proliferative potential of CIS cells together with the expression of placental-like alkaline phosphatase (PLAP), a marker of CIS and germ cell cancer. In dysgenetic testes of 4 intersex children proliferating cell nuclear antigen (PCNA) and PLAP were examined immunohistochemically. DNA content was assessed by densitometry of nucleus in Feulgen stained histologic sections. High incidence of aneuploidy (95.1-97.6% of CIS cells with 2.6-6.8c) was found with the predominant DNA pattern of tri- and tetraploidy in children aged 1 to 3 years. The incidence of triploidy (65-78.1% of cells) was similar to the incidence of the expression of PCNA (53.4-62%), what indicates that part of hyperploid germ cells might represent phase S of cell cycle, but the rest of hyperploid cells might represent neoplastic transformation. In turn, germ cells of 8-months-old patient were predominantly diploid with low incidence of PCNA positive cells which indicate that proliferation/neoplastic transformation of abnormal germ cells is significant mostly after 1 year of age in intersex children. The frequency of PLAP expression in CIS cells (3.1-27% of cells) was weakly related to the frequency of aneuploidy what limits the usefulness of PLAP reaction for the detection of CIS cells.     

Effect of sodium butyrate on the expression of genes transduced by retroviral vectors

We have studied the effects of sodium butyrate (NaBu) on the expression of genes transduced by retroviral vectors and stably expressed in two salivary gland-derived cell lines, A5-DAP and A5-BAG, established earlier. These cell lines were obtained by infecting A5 cells with the retroviral vectors DAP and BAG, respectively, and by selecting neomycin-resistant transduced cells. A5-DAP cells express human placental alkaline phosphatase (PLAP) and A5-BAG cells bacterial β-galactosidase, both under the control of the viral long terminal repeat (LTR) enhancer-promoter. NaBu in the concentration of 2–8 mM inhibited the growth of A5-DAP cells, and induced the expression of heat-stable PLAP. These effects of NaBu were dose-dependent. Induction of PLAP in clones of A5-DAP cells that express different basal levels of the enzyme was not correlated with the relative inducibilty by NaBu. Exposure to 4 mM NaBu for 48 h increased the PLAP mRNA level by 31%. A5-DAP cells released, in a time-dependent manner, PLAP into the culture medium. Cells treated with NaBu released more PLAP than untreated cells in proportion to their elevated level of the enzyme. The parent A5 cells also express a low level of tissue non-specific type alkaline phosphatase, which was also induced by NaBu. NaBu inhibited the growth of A5-BAG cells also, and increased the β-galactosidase level. These data indicate the genes transduced by retroviral vectors can be induced by NaBu, which most likely interacts with the viral LTR. J. Cell. Biochem. 69:201–210, 1998. © 1998 Wiley-Liss, Inc.     

Attachment of human placental-type alkaline phosphatase via phosphatidylinositol to syncytiotrophoblast and tumour cell plasma membranes

Phosphatidylinositol anchors human placental-type alkaline phosphatase (PLAP) to both syncytiotrophoblast and tumour cell plasma membranes. PLAP activity was released from isolated human placental syncytiotrophoblast plasma membranes and the surface of tumour cells with a phospholipase C from Bacillus cereus. This was a specific event, not the result of proteolysis or membrane perturbation, but the action of a phosphatidylinositol-specific phospholipase C in the preparation. Soluble PLAP, released with B. cereus phospholipase C and purified by immunoaffinity chromatography, ran on SDS-PAGE as a 66-kDa band. This corresponded to intact PLAP molecules. The protease bromelain cleaved lower-molecular-mass PLAP (64 kDa) from the membranes. Flow cytometry demonstrated that B. cereus phospholipase C released human tumour cell membrane PLAP in preference to other cell-surface molecules. This was in contrast to the non-specific proteolytic action of bromelain or Clostridium perfringens phospholipase C, which had no effect on membrane PLAP expression. Radiolabelling of tumour cells with fatty acids indicated PLAP to be labelled with both [3H]myristic and [3H]palmitic acid. This fatty-acid--PLAP bond was sensitive to pH 10 hydroxylamine treatment indicating an O-ester linkage.     

The mechanism of placental alkaline phosphatase induction in vitro

The mechanism of placental alkaline phosphatase (PLAP) induction by prednisolone in a uterine cervical epidermoid cancer cell line SKG-IIIa was investigated in vitro by enzyme-cytochemistry, enzyme immunoassay, Northern and Southern blot analysis, and in situ hybridization. Enzyme-cytochemical alkaline phosphatase (ALP) staining and immunoassay revealed increased levels of PLAP (heat-stable ALP) in prednisolone-treated cells. Northern blot analysis and in situ hybridization showed increased amounts of PLAP mRNA. Southern blot analysis indicated that PLAP was not a product of an amplified or rearranged gene. These findings suggest that the induction of PLAP mRNA in SKG-IIIa cells by prednisolone in turn increased the levels of PLAP.     

嫌色性肾细胞癌17例临床病理学分析

目的探讨嫌色性肾细胞癌的临床病理特征、诊断与鉴别诊断要点。方法对17例嫌色性肾细胞癌进行组织形态学、免疫组化染色及Hale’s胶样铁染色观察,结合文献对其临床表现、病理形态特点及鉴别诊断进行探讨。结果嫌色性肾细胞癌17例,大体肿瘤直径3-10.5cm。镜下肿瘤由嫌色细胞和嗜酸细胞构成,呈片状、梁状和腺泡状分布。嫌色细胞体积较大,多角形,胞膜清晰,胞质半透明细网状,胞核皱缩,可见核沟及核异型,核仁不明显;而嗜酸细胞胞质嗜酸,可见明显的核周空晕。免疫组化:EMA 100%阳性,CD10 52.9%阳性,Vimentin阴性,CK7 88.2%阳性,P504S29.4%阳性,CD11794.1%阳性。Hale’s胶样铁染色100%阳性。17例中12例随访6个月到3年,仅1例在术后15个月发现肝脏转移,其余均未发现复发及转移。结论嫌色性肾细胞癌是一种少见的肾肿瘤,恶性程度相对较低,预后良好。掌握该肿瘤独特的病理学特征,对鉴别其他肾上皮性肿瘤有重要帮助。