Morphometrical changes of the human uterine tubes according to aging and menstrual cycle

Studies on the morphological changes in the human uterine tube according to aging and menstrual cycle so far have been limited to microscopic aspects such as cellular changes, mainly due to the inaccessibility of specimens. In this study, postmortem analysis using both macroscopic and microscopic methods was performed using 55 human uterine tubes.The numbers and the degrees of convolution, and the length of the uterine tube had a tendency to decrease according to the increase of age in women by fifties. Under the influence of menopause, the total areas of the tube, mucosal layer and lumen in the ampulla, and lumen in the isthmus and infundibulum were shown to decrease on cross section. However, in the isthmus and infundibulum, the cross sectional area of the tube and mucosal layer did not show statistically significant changes.In the women at reproductive stages, the cross sectional areas of the tube, mucosa and lumen showed variations among different individuals, which may be due to the influence of menstrual cycle rather than the increase of age. No venous engorgement of the tubes was observed at the early proliferative, the mid-secretory and the postmenopausal stage. By contrast, full engorgement was observed at the early secretory stage and the menstrual stage.     

人胚神经干细胞的分离、培养与鉴定

为了探讨人胚神经干细胞体外培养条件和分化情况，摸索出一种切实可行的获得较纯、多潜能人 胚神经干细胞的方法。我们取三月龄人胎脑，用胰蛋白酶消化法分离单个细胞，部分冻存，另一部分进行细胞培养，加EGF、bFGF刺激生长，有限稀释法将获得单细胞克隆，血清诱导分化，并用免疫组化方法进行鉴定。结果显示,EGF和bFGF同时存在的无血清培养基中有大量神经干细胞团生成，含血清培养基则诱导神经干细胞分化成为神经元、星型胶质细胞、少 突胶质细胞。这表明，神经干细胞的存活和分裂有赖于EGF和bEGF的共同作用。经冻存后的胎脑细胞同样能分离培养出有活性的神经干细胞。     

人滋养层细胞侵袭力分子调节机制研究

胎盘是妊娠过程中形成的暂时性的特异器官,它在母体和胎儿间起着重要的桥梁作用。滋养层细胞具有类似于肿瘤细胞迁移和浸润的能力,作为胎盘组织的主要组成细胞之一,在胚胎植入、胎盘的形成和发育等许多生理过程中发挥着重要的作用,但同时也是多种毒素和病原微生物入侵时的靶细胞。滋养细胞侵入过度将导致绒毛膜癌等疾病的发生,浸润不足则可能造成流产、子痫前期等妊娠期疾病。目前,诸多研究表明在胎盘形成过程中,有许多分子和信号通路参与对其滋养层细胞的调控,但滋养层细胞迁移与浸润的具体机制尚不完全明确。本文对人滋养层细胞侵袭力分子调节机制进行了系统论述,旨在为研究病理性妊娠的发生、发展过程提供参考。     

Human Helper T Cell Lines Established by Coculture of Normal Human Cord Leukocytes with an HTLV-Il-infected Rabbit Leukocyte Cell Line (Ra-IIA)

Three helper T cell lines, designated CR -IIA (CR-IIA-1, CR-IIA- 2, and CR-IIA-3), were established by coculturing nor mal human cord leukocytes with a lethally irradiated HTLV II (human T lymphotropic virus type II)- infected rabbit leukocyte cell line (Ra-IIA). CR IIA had a normal human karyotype and expressed the surface markers CD3(+), CD4(+), CD8(-), CD19(-), CD25(+) and HLA- DR(+), confirming their helper T cell nature. CR- IIA cells were all free of Epstein- Barr virus nuclear antigen and were im-muno reactive with serum samples from HTLV- I- or HTLV-Il- infected patients and with anti HTLV- I, p19 or p24 anti body. The provirus genome of HTLV-II was detected in these cell lines by the polymerase chain reaction combined with a digoxigenin- enzyme- linked immunosorbent assay. Electron microscopy of CR-IIA-1 cells revealed a few im mature type C virus particles. These results suggest that HTLV- II was transmitted from the infected rabbit leukocyte cell line to human cord helper T lymphocytes with the development of immortalized HTLV - II- producing T cell lines. Acta Pathol Jpn 42: 347–352, 1992.     

Immunohistochemical Demonstration of Cell Proliferation and Estrogen Receptor Status in Human Breast Cancer

Cell proliferation and estrogen receptor (ER) status was investigated in 45 invasive ductal carcinomas of the breast by immunohistochemical methods using monoclonal antibodies Ki 67 (anti-human proliferating cell antibody) and ER ICA. The results were assessed on the basis of nuclear staining intensity and the percentage of positively stained tumor cell nuclei (index score). There was a significant inverse correlation between the Ki 67 and ERICA index scores, although 4 cases showed high index scores for both markers. We conclude that ER positive cells do not always have low proliferation activity, which may be one of the reasons why endocrine therapy is not effective against all ER positive breast cancers. Acta Pathol Jpn 40: 902 907, 1990.     

Date Seed Oil Inhibits Hydrogen Peroxide-Induced Oxidative Stress in Normal Human Epidermal Melanocytes

The administration of antioxidants has been shown to enhance repair and healing processes in cutaneous tissue. Date seed oil (DSO) extract, which might be a potential source of natural antioxidants such as phenols and tocopherols, has been reported to be beneficial in the reduction of chemically induced oxidative stress in normal human skin. In this study, we investigated the protective effects of DSO against hydrogen peroxide (H₂O₂)-induced oxidative stress in terms of lipid peroxidation, depletion of such endogenous antioxidant defense enzymes as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) using normal human epidermal melanocytes (NHEM). The results showed that DSO, endowed with a radical scavenging ability, decreased oxidative injury by inhibition of damage caused by H₂O₂. Treatment of NHEM with DSO inhibited H₂O₂-induced lipid peroxidation. In addition, the extract inhibited H₂O₂-induced depletion of antioxidant defense components, such as SOD, CAT, and GPx. Our findings demonstrate that DSO is an efficient extract able to prevent melanocytes oxidative damage induced by H₂O₂ exposure. Thus it may be a potential promising candidate, as a chemopreventive agent, in the development of melanocyte-related pathologies like vitiligo and melanoma.     

Changes in Morphology and Cellular Replacement of Surface Epithelia in Human Gastric Mucosa Affected by Different Pathological Changes: A Scanning Electron Microscopical Study of Human Gastric Biopsy Material

The surface epithelium of normal gastric mucosa shows a constant turnover, migrating from the proliferation zone at the neck of the gastric glands to the mucosal surface. The cytologic changes associated with the concomitant cellular differentiation were so far studied in sections by light and electron microscopy only. With these methods neither the changes of surface morphology associated with cellular maturation nor the shedding of surface epithelia can be recorded. With the scanning electron microscope it is possible to study especially these changes of the mucosal surface plus those of the single surface epithelia over a wide range of magnification and with a sufficient depth of focus.In this study therefore an attempt was made to correlate some scanning electron microscopic findings concerning these changes of mucosal and cellular surface with the results of parallel light microscopic examinations. For this purpose double biopsies of human gastric mucosa were taken during routine endoscopic examination. One of these biopsies was processed for light microscopic examination after formaldehyde fixation. The other was fixed in buffered glutaraldehyde solution and 1 % OS04 solution and thereafter the material was processed for scanning electron microscopic study by dehydration against an increasing acetone gradient, critical point drying and sputter coating of the dry specimens which were mounted on metal stubs with conductive silver.Some aspects of surface morphology of the mucosa and the single cells in correlation to different types of gastritis are demonstrated and the morphological expressions of cytological degeneration and shedding from the mucosal surface are commented on.     

Morphometrical analysis of gall-bladder adenoma and adenocarcinoma with reference to histogenesis and adenoma-carcinoma sequence

Summary In order to examine whether our subdivision of gall-bladder adenoma and adenocarcinoma into nonmetaplastic and metaplastic types is reasonable from the viewpoint of their cytological features, a morphometrical analysis was conducted on 17 adenomas and 59 adenocarcinomas. The morphometrical parameters used were nucleo-cytoplasmic ratio (N/C ratio) and nuclear areas (NA). N/C ratio in the metaplastic type of both adenoma and adenocarcinoma was significantly larger than that in the non-metaplastic. This result shows that the different tumour types are associated with a different N/C ratio. Continuous measurement of N/C ratio and NA in progressing from non-cancerous mucosa to the lesion was made and the data obtained were analysed by the Lowess method. In some adenomas the total area of polypoid lesions was serially measured and these data were also analysed by the Lowess method. The results showed different processes in non-metaplastic and metaplastic types of adenoma and adenocarcinoma from the standpoint of nuclear changes of N/C ratio and NA. These results indicate that our histogenetic classification of adenocarcinoma is reasonable in morphometrical nuclear analysis. We also investigated the adenoma-carcinoma sequence as a possible histogenesis for gall-bladder carcinoma. Eight (72.7%) of 11 metaplastic adenomas and two (33.3%) of six non-metaplastic adenomas had foci of atypical gland proliferation and were considered to be carcinomas. Moreover, these carcinomatous areas were surrounded by severe dysplasia. These findings indicate that adenoma-carcinoma sequence accounts for one of the histogeneses of gall-bladder carcinoma.     

Cell Kinetic and morphological studies of human cholangiocellular carcinoma

We investigated the cell kinetics and morphologies of cholangiocellular carcinoma (CCC) using 48 autopsied or surgically resected cases (47 were adenocarcinoma and the remaining adenosquamous cell carcinoma), all of which were formalin-fixed and paraffin-embedded. Cell kinetics were analyzed by counting the number of argyrophilic nucleolar organizer regions (AgNOR) using immunostaining of proliferating cell nuclear antigens (PCNA) and flow cytometric DNA analysis. Dedifferentiation of CCC was positively correlated with AgNOR number (2.22 ± 0.21 in well differentiated, 3.66 ± 0.85 in moderately differentiated and 4.17 ± 0.49 in poorly differentiated adenocarcinomas, respectively). In 22 cases, the labeling index (LI) of PCNA was higher in moderately and poorly differentiated adenocarcinomas (24.0 ± 2.35 and 26.0 ± 4.89, respectively) than in well differentiated ones (10.8 ± 2.14). A majority of well differentiated ones were diploid, while aneuploidy prevailed in moderately to poorly differentiated ones. These data suggest that cell proliferative indices and nuclear DNA analysis of CCC accurately reflect their histological grading. The anatomical location of CCC along the biliary tree had no relation to either of the cell kinetic data. In autopsy cases, the patients with organ and lymph node metastases tended to show a higher DNA index and aneuploidy. This study implies that a combination of several cell kinetic data is valuable for the evaluation of the biological behaviors of CCC, and also supports further studies of cell kinetics of CCC using small-sized biopsy specimens, as a prognostic indicator.     

The Effects of Fructus Xanthii Extract on Cytokine Release from Human Mast Cell Line (HMC-1) and Peripheral Blood Mononuclear Cells

In the present study, human mast cell line (HMC-1) and peripheral blood mononuclear cells (PBMNC) were pre-incubated with various concentrations of Fructus xanthii extract solution followed by being stimulated with phorbol myristate acetate and further incubated for 24 hours. The cytokines, in cell culture supernatants, of interleukin (IL)-4, IL-6, IL-8, GM-CSF, and TNF-α in the supernatant were measured by enzyme-linked immunosorbent assay. The results demonstrated that Fructus Xanthii could modulate the mast cell-mediated and PBMNC-mediated inflammatory and immunological reactions modulated by cytokines. And our study supplied the evidence of the mechanisms of Fructus Xanthii in treating inflammatory and allergic diseases.     

Human B Cell Biology

Human B lymphocytes share one major distinctive feature with B cells of other higher animals, namely the ability to generate and secrete immunoglobulins. These highly specialized proteins are capable of tremendous diversity, and thereby account for much of our immune protection against invading organisms. Despite the great potential diversity possible in the specificities of immunoglobulin molecules, however, the binding of antibody to antigen initiates a limited spectrum of biologically important effector functions, such as complement activation and/or adherence of the immune complex to receptors on leukocytes. A variety of mechanisms have been elucidated that account for this, not all of which are shared by the different types of animals capable of making these proteins. The purpose of this chapter is to review the genetic, developmental, and physiologic mechanisms critical for human B cell expression of immunoglobulin.     

重组人血管内皮抑素对人多发性骨髓瘤细胞RPMI 8226增殖和相关蛋白表达的影响

 目的： 研究重组人血管内皮抑素（恩度）在体外对人多发性骨髓瘤细胞株RPMI 8226细胞增殖、细胞周期及细胞相关蛋白表达的影响。方法： 用CCK-8检测恩度对RPMI 8226细胞增殖的影响;流式细胞术检测凋亡和细胞周期的改变;Western blotting检测凋亡相关蛋白Bcl-2和caspase-3表达变化;以实时定量PCR和Western blotting检测RPMI 8226细胞血管细胞粘附因子 1（VCAM-1）、白细胞介素 6（IL-6）和血管内皮生长因子（VEGF）mRNA以及蛋白表达变化;ELISA检测细胞上清液中细胞因子IL-6和VEGF水平。结果： 恩度对RPMI 8226细胞有增殖抑制作用,250 mg/L恩度对RPMI 8226细胞 72 h增殖抑制率为（59.5±5.6）%（P<0.05）,细胞G1期比例增加（P<0.05）,但对细胞凋亡及凋亡相关蛋白Bcl-2和caspase-3表达无显著影响;经恩度作用后RPMI 8226细胞 VCAM-1表达下降（P<0.05）,IL-6和VEGF表达及分泌降低（均P<0.05）。结论： 恩度能通过改变细胞周期变化抑制RPMI 8226细胞增殖,并能减少VCAM-1表达及IL-6、VEGF分泌,对细胞凋亡无明显改变。     

Effects of maternal bilateral ureteral ligation on fetal development of kidney in rats: Morphometrical changes in glomerular components

Development of the glomerulus in fetal kidney was studied morphometrically following ligation of both ureters of pregnant rats. The ligation was performed on days 17, 19, and 21 of gestation, and autopsy was done 24 hr after each operation. The percentage volumes of the five glomerular components (epithelial cells, capillary, mesangium, glomerular basement membrane, and Bowman's space) were determined by point counting, and the surface area of glomerular basement membrane per unit volume of glomerulus was calculated by intercept counting. On fetal days 18, 20, and 22, percentage volume of Bowman's space in the fetuses from the ligated mothers was significantly smaller than that in the fetuses from sham-ligated mothers. On fetal days 20 and 22, surface area of glomerular basement membrane and glomerular capillary length per unit volume of glomerulus and percentage of glomerular capillary volume in the fetuses from the ligated mothers were significantly larger than those in the fetuses from sham-ligated mothers. These results suggest that maternal bilateral ureteral ligation induces an increase in filtration area of glomerulus in fetal kidney when the fetal kidney is functional. © 1993 Wiley-Liss, Inc.     

Expression of Leukocyte Common Antigen (CD45) on Various Human Leukemia/Lymphoma Cell Lines

CD45 antigen (leukocyte common antigen), a unique and ubiquitous membrane glycoprotein with a molecular mass of about 200 kDa, is expressed on almost all hematopoietic cells except for mature erythrocytes. However, the biological function of this glycoprotein still remains to be resolved. In order to clarify the role of CD45 antigen in hematopoietic cell differentiation and function, its expression on human leukemia lymphoma cell lines was studied by membrane immunofluorescence. Thirty eight established cell lines were analyzed using T29–33, a monoclonal antibody (MoAb) that recognizes the common epitopes of this glycoprotein molecule. Conventional cell marker studies were also carried out on these cell lines to compare their CD45 expression. It was shown that CD45 expression varies among B lineage cells depending on cell differentiation, in contrast to its stable expression on leukemic T cell (6/6, positive) and myeloid (5/5, positive) lineage cell lines. On the other hand, only two out of six histiomonocytoid lineage cell lines were positive. Human T cell leukemia lymphoma virus type I (HTLV-I) associated T cell lines derived from peripheral blood leukocytes of patients with adult T cell leukernia/ lymphoma (ATL/L) in Japan did not express CD45 on their cell surface. Taken together, these observations suggest that CD45 has a functional role in hematopoietic cell activation and differentiation. Acta Pathol Jpn 40: 107–115, 1990.     

Colorectal mucin histochemistry in health and disease: A critical review

Neoplastic, inflammatory and regenerative processes affecting colorectal mucosa are associated with alterations in structure of epithelial mucin. This review collates mucin-, lectin-, and immuno-histochemical observations on colorectal mucins and introduces recent molecular genetic insights into the structure of the protein backbone of mucins. The numerous structural modifications uncovered by the various technical approaches have been reduced to a few manageable principles that are of relevance to both researcher and diagnostic pathologist. Particular attention is drawn to the need to appreciate the limited specificities of probes, the confounding influences of anatomical site and genetic factors (necessitating the use of appropriate positive and negative control tissues) and the precise location of secretory material. In the past, insufficient attention has been given to the effects of altered differentiation including metaplasia and differing lineage expression in epithelial disorders of growth. It is likely that certain changes loosely ascribed to goblet cell mucin, such as neo-expression of blood group antigens and anomalous expression of core carbohydrate structures, do not occur at all. Critical examination of available data point to only two consistent and unequivocal changes affecting goblet cell mucin in pathological processes: loss of O-acetyl substituents at sialic acid C₄ and C_7,8,9 and increased sialyla-tion. Furthermore, there are no neoplasia-specific alterations in mucins documented to date. All neoplasia-associated changes have been described in non-neoplastic lesions also.     

Comparative Phenotypic Studies of Duct Epithelial Cell Lines Derived from Normal Human Pancreas and Pancreatic Carcinoma

We have investigated the mRNA/protein expression of several tyrosine kinase receptors, growth factors, and p16^INK4A cyclin inhibitor in cell lines derived from normal human pancreatic duct epithelium (HPDE) and compared them with those of five pancreatic ductal carcinoma cell lines. Cultured HPDE cells express low levels of epidermal growth factor receptor (EGFR), erbB2, transforming growth factor (TGF)-α, Met/hepatocyte growth factor receptor (HGFR), vascular endothelial growth factor (VEGF), and keratinocyte growth factor (KGF). They also expressed high levels of amphiregulin but did not express EGF and cripto. The expression levels were similar in primary normal HPDE cells and those expressing transfected E6E7 genes of human papilloma virus-16, but their immortalization appeared to enhance the expression of EGFR and Met/HGFR. In comparison, pancreatic carcinoma cell lines commonly demonstrated overexpression of EGFR, erbB2, TGF-α, Met/HGFR, VEGF, and KGF, but they consistently showed marked down-regulation of amphiregulin mRNA expression. In contrast to all carcinoma cell lines that showed deletions of the p16 gene, HPDE cells consistently demonstrated normal p16 genotype and its mRNA expression. This is the first report that compares the phenotypic expression of cultured pancreatic ductal carcinoma cells with epithelial cell lines derived from normal human pancreatic ducts. The findings confirm that malignant transformation of human pancreatic duct cells commonly results in a deregulation of expression of various growth factors and receptors.     

甘草酸对人肺癌细胞增殖和侵袭抑制作用的机制探讨

目的　探讨甘草酸对高转移人肺癌细胞 (PGCL3)增殖抑制、抗侵袭和诱导凋亡的作用 ,并探讨其抗增殖和侵袭作用的机制 .方法　用 0 .5mmol/L和 1.0mmol/L甘草酸处理PGCL3细胞 96h ,进行细胞增殖抑制试验、软琼脂集落形成试验、侵袭、粘附和运动试验 ,以及组织蛋白酶B活性的测定 ,观察药物对PGCL3细胞增殖和侵袭能力的影响 .上述两浓度处理细胞 4 8h ,用吖啶橙 -溴化乙锭荧光双染法和末端脱氧核苷酸转移酶标记法 (TUNEL法 )检测细胞凋亡 .结果　甘草酸使PGCL3细胞增殖抑制率升高 (p <0 .0 5和p <0 .0 1)和软琼脂集落形成率显著下降(p <0 .0 1) ,呈剂量依赖性 ,半增殖抑制浓度 (IC50 )为 1.8mmol/L ;甘草酸明显减弱细胞侵袭能力 (p <0 .0 1) ,涉及侵袭关键环节的细胞粘附、运动和组织蛋白酶B分泌均受明显抑制 (p <0 .0 5和p <0 .0 1) ;甘草酸还使细胞凋亡率显著升高 (p<0 .0 1) .结论　甘草酸具有抑制PGCL3人肺癌细胞增殖和侵袭作用 .其抑制增殖的机制是通过对细胞分化和凋亡的诱导作用 ;其抗侵袭机制是对侵袭的多个基本环节起抑制作用 .     

Cell degeneration and mitosis in the buccopharyngeal and branchial membranes in the mouse embryo

Summary The frequencies of cell degeneration and mitosis were investigated in the rupturing buccopharyngeal membrane (BPM) and in the persistent first branchial membrane (BM). In the BPM, cell degeneration starts many hours before rupture is visible, but mitotic figures are absent. In the BM this situation is reversed: mitotic figures are regularly observed, but a degenerating cell only occasionally. It is concluded that the ratio between the numbers of degenerating and dividing cells regulates the fate of both the BPM and the BM.     

In Situ Localization of Pepsinogens I and II mRNA in Human Gastric Mucosa

Localization of pepsinogens I and II mRNA in the human gastric mucosa was investigated by an in situ hybridization method using digoxigenin labeled cDNA probes. Gastric fundic mucosa from healthy volunteers, which was stained with digoxigenin labeled pepsinogens I and II cDNA probes, showed positive staining in the cytoplasm of both chief cells and mucous neck cells. In contrast, gastric antral mucosa stained with the pepsinogen I cDNA probe showed no positive reaction in the surface mucous cells or pyloric glands. On the other hand, the pyloric glands were stained positively with the pepsinogen II cDNA probe and the staining appeared to be identical to that obtained with the anti pepsinogens I and II monoclonal antibodies using the avidin biotin peroxidase complex technique. These results are consistent with those of previous studies that have employed immunochemical and immunohistochemical techniques. Acta Pathol Jpn 39: 765 771, 1989.