Clinical Observations EFFECT OF THE COMBINATION OF CHOLESTEROL AND BILE ACID ON THE METABOLISM OF CHOLESTEROL，PHOSPHOLIPS AND

ＣｌｉｎｉｃａｌＯｂｓｅｒｖａｔｉｏｎｓＥＦＦＥＣＴＯＦＴＨＥＣＯＭＢＩＮＡＴＩＯＮＯＦＣＨＯＬＥＳＴＥＲＯＬＡＮＤＢＩＬＥＡＣＩＤＯＮＴＨＥＭＥＴＡＢＯＬＩＳＭＯＦＣＨＯＬＥＳＴＥＲＯＬ，ＰＨＯＳＰＨＯＬＩＰＳＡＮＤＢＩＬＥＡＣＩＤＩＮＨＥＰＡＴＯ...     

A MOLECULAR EPIDEMIOLOGIC MARKER OF HEPATOCELLULAR CARCINOMA FROM AFLATOXIN B1 CONTAMINATED AREA IN THE SOUTHWEST OF GUANGXI

ＡＭＯＬＥＣＵＬＡＲＥＰＩＤＥＭＩＯＬＯＧＩＣＭＡＲＫＥＲＯＦＨＥＰＡＴＯＣＥＬＬＵＬＡＲＣＡＲＣＩＮＯＭＡＦＲＯＭＡＦＬＡＴＯＸＩＮＢ１ＣＯＮＴＡＭＩＮＡＴＥＤＡＲＥＡＩＮＴＨＥＳＯＵＴＨＷＥＳＴＯＦＧＵＡＮＧＸＩＤｅｎｇＺｈｕｏｌｉｎ邓卓霖ＭａＹ...     

WHOLE ORGAN SUBSERIAL SECTION EXAMINATIONOF OCCULT BREAST CARCINOMA

ＷＨＯＬＥＯＲＧＡＮＳＵＢＳＥＲＩＡＬＳＥＣＴＩＯＮＥＸＡＭＩＮＡＴＩＯＮＯＦＯＣＣＵＬＴＢＲＥＡＳＴＣＡＲＣＩＮＯＭＡＮｉｕＹｕｎ牛昀；ＦｕＸｉｌｉｎ傅西林；ＦａｎＹｕ范宇；ＦａｎｇＺｈｉｙｉ方志沂（ＴｉａｎｊｉｎＣａｎｃｅｒＨｏｓｐｉｔａｌａｎｄ...     

THE BLOCKING EFFECTS OF GLYCYRRHIZE URALENSIS AND CHELIDONIUM MAJUS ON MUTAGENESIS INDUCED BY AFLATOXIN B1

ＴＨＥＢＬＯＣＫＩＮＧＥＦＦＥＣＴＳＯＦＧＬＹＣＹＲＲＨＩＺＥＵＲＡＬＥＮＳＩＳＡＮＤＣＨＥＬＩＤＯＮＩＵＭＭＡＪＵＳＯＮＭＵＴＡＧＥＮＥＳＩＳＩＮＤＵＣＥＤＢＹＡＦＬＡＴＯＸＩＮＢ１ＳｈｉＧｕｉｚｎｉ；史桂芝；ＪｉＸｉｎｈｕａ；纪新华；ＬｉａｎｇＹ...     

PRINCIPLES AND APPLICATIONS OF MOLECULAR EPIDEMIOLOGY AND STUDIES OF HUMAN PROSTATIC CARCINOMA

ＰＲＩＮＣＩＰＬＥＳＡＮＤＡＰＰＬＩＣＡＴＩＯＮＳＯＦＭＯＬＥＣＵＬＡＲＥＰＩＤＥＭＩＯＬＯＧＹＡＮＤＳＴＵＤＩＥＳＯＦＨＵＭＡＮＰＲＯＳＴＡＴＩＣＣＡＲＣＩＮＯＭＡＴｚｅ－ｃｈｅｎＨｓｉｅｈ；ＣｈｕｎｙｕｎｇＮｇａｎｄＪｏｓｅｐｈＭ．Ｗｕ（Ｄｅｐｔ...     

THE IN VITRO POTENTIATION OF LAK CELL CYTOTOXICITY IN CANCER AND AIDS PATIENTS INDUCED BY F3—A FRACTIONATED EXTRACT OF ASTRAG

ＴＨＥＩＮＶＩＴＲＯＰＯＴＥＮＴＩＡＴＩＯＮＯＦＬＡＫＣＥＬＬＣＹＴＯＴＯＸＩＣＩＴＹＩＮＣＡＮＣＥＲＡＮＤＡＩＤＳＰＡＴＩＥＮＴＳＩＮＤＵＣＥＤＢＹＦ３—ＡＦＲＡＣＴＩＯＮＡＴＥＤＥＸＴＲＡＣＴＯＦＡＳＴＲＡＧＡＬＵＳＭＥＭＢＲＡＮＡＣＥＵＳＣｈｕ...     

LOCALIZATION OF TYPE I AND TYPE Ⅲ PROCOLLAGEN mRNAs IN BREAST SCIRRHOUS CARCINOMA BYIN SITU HYBRIDIZATION

ＬＯＣＡＬＩＺＡＴＩＯＮＯＦＴＹＰＥＩＡＮＤＴＹＰＥⅢＰＲＯＣＯＬＬＡＧＥＮｍＲＮＡｓＩＮＢＲＥＡＳＴＳＣＩＲＲＨＯＵＳＣＡＲＣＩＮＯＭＡＢＹＩＮＳＩＴＵＨＹＢＲＩＤＩＺＡＴＩＯＮＧａｎＹｕｅｔｏ；干月波；ＺｎｅｎｇＳｈｕ；郑树；ＹｕＨａｉ；余海（Ｃ...     

RETINOIC ACID NUCLEAR RECEPTORα （RARα），A MAJORROLE IN MEDIATING RETINOIDS INHIBITION OF GROWTH IN HU

ＲＥＴＩＮＯＩＣＡＣＩＤＮＵＣＬＥＡＲＲＥＣＥＰＴＯＲα（ＲＡＲα），ＡＭＡＪＯＲＲＯＬＥＩＮＭＥＤＩＡＴＩＮＧＲＥＴＩＮＯＩＤＳＩＮＨＩＢＩＴＩＯＮＯＦ　ＧＲＯＷＴＨＩＮＨＵＭＡＮＢＲＥＡＳＴＣＡＲＣＩＮＯＭＡＣＥＬＬＳＳｈａｏＺｈｉｍｉｎ邵志敏；...     

STUDY ON ANTITUMOR DRUG-INDUCED APOPTOSIS IN HUMAN CANCER CELLS BY TERMINAL DEOXYNU-CLEOTIDYL TRANSFERASE ASSAY

ＳＴＵＤＹＯＮＡＮＴＩＴＵＭＯＲＤＲＵＧＩＮＤＵＣＥＤＡＰＯＰＴＯＳＩＳＩＮＨＵＭＡＮＣＡＮＣＥＲＣＥＬＬＳＢＹＴＥＲＭＩＮＡＬＤＥＯＸＹＮＵＣＬＥＯＴＩＤＹＬＴＲＡＮＳＦＥＲＡＳＥＡＳＳＡＹＴｏｎｇＴｏｎｇ童彤ＳｕｎＨａｎｘｉａｏ孙含笑Ｌｉｕ...     

THE EXPRESSION OF C－MYC AND N－RAS ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA－AN IN SITU HYBRIDIZATION STUDY ON PARAFFIN EMBE

ＴＨＥＥＸＰＲＥＳＳＩＯＮＯＦＣ－ＭＹＣＡＮＤＮ－ＲＡＳＯＮＣＯＧＥＮＥＳＩＮＨＵＭＡＮＨＥＰＡＴＯＣＥＬＬＵＬＡＲＣＡＲＣＩＮＯＭＡ－ＡＮＩＮＳＩＴＵＨＹＢＲＩＤＩＺＡＴＩＯＮＳＴＵＤＹＯＮＰＡＲＡＦＦＩＮＥＭＢＥＤＤＥＤＴＩＳＳＵＥＳＥＣＴＩＯＮ...     

Differences in surface sialic acid and galactosyl residues of two autologous human melanoma cell lines

Cell surface carbohydrate differences were observed between two human cell lines initiated from primary melanoma and metastasis of the same patient. Although total sialic acid content was similar in both cell lines, neuraminidase-released sialic acid was twice as high in metastatic cells than that of primary cells. One class of Concanavalin A binding sites with similar affinity constant was found in untreated and neuraminidase-treated cells in both cell lines. Before surface sialic acid release, the primary cell line expressed two classes of Ricinus lectin binding sites with high and low affinity; the cell line of metastatic origin had only one class of Ricinus lectin binding sites with low affinity. After neuraminidase treatment, the number of Ricinus lectin binding sites with low affinity increased two- or three-fold in both cell lines, whereas the high-affinity binding sites were not observed in primary cells. The present data indicated that differences in surface sialic acid level modified the Ricinus lectin binding in two human melanoma cell lines. However, the ability of the cells to bind Concanavalin A was not changed.     

Wheatgerm agglutinin-mediated toxicity in pancreatic cancer cells.

Lectin binding specificities for carbohydrate allow phenotypic and functional characterization of membrane-associated glycoproteins expressed on cancer cells. This analysis examined wheatgerm agglutinin binding to pancreatic cancer cells in vitro and the resulting toxicity. Membrane preparations of nine human pancreatic carcinoma cell lines were studied for lectin binding using wheatgerm agglutinin (WGA), concanavalin A (ConA) and phytohaemagglutinin-L (PHA-L) in a lectin blot analysis. Cell proliferation in vitro was measured by thymidine incorporation in the absence or presence of lectins at various concentrations. Sialic acid binding lectins or succinyl-WGA (succWGA) served as controls. WGA toxicity was tested after swainsonine or neuraminidase pretreatment. Binding and uptake of fluorescein-labelled lectins was studied under fluorescence microscopy. All pancreatic cell lines displayed high WGA membrane binding, primarily to sialic acid residues. Other lectins were bound with weak to moderate intensity only. Lectin toxicity corresponded to membrane binding intensity, and was profound in case of WGA (ID50 at 2.5-5 microg ml(-1)). WGA exposure induced chromatin condensation, nuclear fragmentation and DNA release consistent with apoptosis. Important steps for WGA toxicity included binding to sialic acid on swainsonine-sensitive carbohydrate and lectin internalization. There was rapid cellular uptake and subsequent nuclear relocalization of WGA. In contradistinction to the other lectins studied, WGA proved highly toxic to human pancreatic carcinoma cells in vitro. WGA binding to sialic acid residues of N-linked carbohydrate, cellular uptake and subsequent affinity to N-acetyl glucosamine appear to be necessary steps. Further analysis of this mechanism of profound toxicity may provide insight relevant to the treatment of pancreatic cancer.     

Regeneration of membrane sialic acid after neuraminidase treatment of leukemic granulocytes

Granulocytes from patients with chronic myelogenous leukemia (CML) were studied for their ability to regenerate surface sialic acid following treatment with Vibrio cholera neuraminidase (VCN) in vitro. Immediately after neuraminidase treatment, CML and normal granulocytes showed similar incorporation of radioactivity after surface labelling with sodium periodate/potassium-H3-borohydride (PI/BH3(4)). CML granulocytes treated with neuraminidase then incubated for 18 h in nutrient medium showed strikingly increased PI/BH3(4) labelling, usually greater than initial pretreatment values, consistent with a rapid reappearance of sialic acid in the cell membrane. This pattern was not seen in normal granulocytes. The aberrant regeneration of sialic acid in CML granulocytes in vitro could be inhibited by addition of 3 X 10(-6) M retinoic acid, suggesting either a direct effect on membrane glycoconjugate synthesis or an association with granulocyte differentiation.     

Expression of cell surface glycoproteins in human melanoma cell lines with different tumorigenic properties

Human malignant melanoma cell lines characterized by either a high or a low ability to grow subcutaneously in athymic nude mice have been examined for their cell-surface glycoproteins. Striking differences were demonstrated between these 2 groups. Cells from lines of low tumorigenicity (LT group) displayed twice as much Vibrio cholerae neuraminidase and galactose oxidase accessible glycoproteins as cells from lines of high tumorigenicity (HT group) and each group of cell lines could be characterized by specific glycoprotein profiles. LT and HT group cells displayed similar amounts of periodate accessible glycoproteins, but sialoglycoprotein profiles were characteristic for each group of cell lines. Furthermore, whereas 87% of the sialic acid released by V. cholerae neuraminidase came from cell surface glycoproteins in HT group cells, only 53-55% of the released sialic acid came from surface glycoproteins in LT group cells. These results suggest that human melanoma cell lines exhibiting different tumorigenicity in nude mice can also be characterized by differences in composition and organization within the plasma membranes of their cell-surface sialoglycoproteins.     

Some effects of neuraminidase on the in vitro interactions between spleen and mastocytoma (P815) cells

A recently advanced generalization is that a sialomucin coat surrounding tumour cells protects them from immunological surveillance. We have therefore studied the effects of neuraminidase (sialidase) on the immunolysis of P815 mastocytoma cells by sensitized spleen cells, in vitro, to test this hypothesis. In a very sensitive system in which immunolysis was quantitated by measurements of the release of ⁵¹Cr from labelled P815 cells, neuraminidase treatment of the target cells did not demonstrably make them more susceptible to attack by spleen cells. Killing proceeded in spite of a substantial contribution of sialic acid moieties to the net negative surface charge of the untreated P815 cells, and was not enhanced by enzymatic removal of substantial amounts of their bound surface sialic acids, as determined by cell electrophoresis. Thus, the generalization cannot be substantiated by our experiments. An interesting positive finding was that neuraminidase treatment of the killer, spleen cell population, enhances the ⁵¹Cr-release from their targets.     

Association of adriamycin-induced resistance to NK-mediated lysis with sialic acid level and immunological reactivity of transferrin receptors and glycophorin A

Adriamycin (ADM) can increase sialic acid content in K 562 cells and reduce their susceptibility to NK-mediated lysis. In this report, hypothetical relationship between this resistance and augmentation in sialylation has been investigated. Variations in the time of exposure to ADM showed that 12 hours were sufficient to cause maximal recruitment of benzidine-positive cells, growth inhibition and resistance to NK-mediated lysis. On the contrary, the membrane sialic acid density seemed stable and 24 hours of drug exposure were necessary to observe a clear rise in sialic acid. Neuraminidase treatment of control and ADM-treated K 562 cells was associated with an obvious enhancement in their susceptibility to NK-mediated lysis which can be explained by an increase in the target-effector binding ability as assessed by a direct conjugate-forming cell assay. However, the neuraminidase treatment did not modify the sensitivity difference to lysis between untreated and ADM-treated cells. As compared to control the reactivity of ADM-treated cells was higher with an antiglycophorin A (GPA) MAb and lower with an antitransferrin receptor (TFR) MAb. Kinetic studies suggested that GPA expression is a better index of ADM-induced resistance to NK-mediated lysis than TFR expression. In addition, neuraminidase treatment showed that TFR and GPA modulations induced by ADM can be correlated with sialylation alterations.     

Lectin histochemistry of normal and neoplastic nasopharyngeal epithelium.

The cell surface carbohydrate profile of formalin-fixed paraffin-embedded tissue sections of normal and neoplastic epithelium was evaluated using 9 plant lectins. Three lectins, namely Con A, RCA and WGA, showed a similar pattern and staining intensity from normal epithelium to metaplastic squamous epithelium and nasopharyngeal intraepithelial neoplasia (NPIN). However, a decrease in staining reactivity was observed in undifferentiated nasopharyngeal carcinoma. Significant differences in intensity and distribution were seen in UEA and cryptic PNA residue (after neuraminidase pretreatment) from normal nasopharyngeal epithelium to NPIN. Infiltrative undifferentiated carcinomas showed a heterogenous lectin binding pattern and altered intensity of lectin binding in one case of DBA and three cases of PNA (no neuraminidase pretreatment), suggesting a variation in expression of carbohydrate by tumour cells. These results indicate that neoplasia in nasopharyngeal epithelium is associated with alterations in terminal sialic acid, -Fucose residues and -Gal-D-GalNac residues present in the outer parts of glycoconjugates. SBA, VVL and BSL failed to stain any types of epithelia. Desialylation of tissues by preincubation with neuraminidase did not expose DBA, SBA, VVL and BSL binding sites. These findings may be used as a baseline for evaluation of lectin binding in preinvasive and invasive lesions of the nasopharynx.     

Muscarinic receptor coupling to intracellular calcium release in rat pancreatic acinar carcinoma

Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cholinergic receptor protein affinity labeled with the muscarinic antagonist [3H]propylbenzilylcholine mustard revealed a major polypeptide with molecular weight of 80,000-83,000 in both acinar carcinoma and normal acinar cells of rat pancreas. Muscarinic receptor protein is therefore conserved in pancreatic acinar carcinoma. A small but significant difference was detected in the affinity of carcinoma cell receptors (Kd approximately 0.6 nM) and normal cell receptors (Kd approximately 0.3 nM) for reversible binding of the muscarinic antagonist drug, N-methylscopolamine. In addition, carcinoma cell muscarinic receptors displayed homogeneous binding of the agonist drugs carbamylcholine (Kd approximately 31 microM) and oxotremorine (Kd approximately 4 microM), whereas normal cell receptors demonstrated heterogeneous binding, with a minor receptor population showing high affinity binding for carbamylcholine (Kd approximately 3 microM) and oxotremorine (Kd approximately 160 nM), and a major population showing low affinity binding for carbamylcholine (Kd approximately 110 microM) and oxotremorine (Kd approximately 18 microM). Both carcinoma and normal cells exhibited concentration-dependent carbamylcholine-stimulated increases in cytosolic free Ca2+, as measured by 45Ca2+ outflux assay and intracellular quin 2 fluorescence. However, carcinoma cells were observed to be more sensitive to Ca2+ mobilizing actions of submaximal carbamylcholine concentrations, demonstrating 50% maximal stimulation of intracellular Ca2+ release at a carbamylcholine concentration (approximately 0.4 microM) approximately one order of magnitude below that seen for normal cells. These results indicate altered muscarinic receptor coupling to intracellular Ca2+ release in acinar carcinoma cells, which manifests as a single activated receptor state for agonist binding, and increased sensitivity of Ca2+ release in response to muscarinic receptor stimulation.     

Sialic acids linked to glycoconjugates of Fas regulate the caspase-9-dependent and mitochondria-mediated pathway of Fas-induced apoptosis in Jurkat T cell lymphoma

To clarify the functions of sialic acids linked to glycoconjugates of Fas in Fas-induced apoptosis, Jurkat T cells, untreated and treated with neuraminidase, were incubated with anti-Fas monoclonal antibody, CH11. Apoptosis of Jurkat T cells induced by incubation with CH11 was enhanced by the pre-treatment with neuraminidase. By flow cytometry sialylated glycoconjugates were detected on the cell surface of Jurkat T cells using LFA lectin, which specifically reacts with sialic acid, and pre-treatment with Vibrio Cholerae neuraminidase resulted in desialylation of Jurkat cell surface glycoconjugates. The enhancement of Fas-induced apoptosis by pre-treatment with neuraminidase was inhibited by z-VAD-fmk, a broad caspase inhibitor, and Ac-LEHD-CHO, an inhibitor of caspase-9, but not by Ac-IETD-CHO an inhibitor of caspase-8 or 6, imipramine, an inhibitor of acidic sphingomyelinase, glutathione, an inhibitor of neutral sphingomyelinase and Fumonisin B1, an inhibitor of ceramide synthase. Mitochondrial membrane potentials (Deltapsim) measured with a Mitocapture assay kit demonstrated that the loss of Deltapsim involved in Fas-induced apoptosis was enhanced by pre-treatment with neuraminidase. Furthermore, Western blot analysis using polyclonal antibody (C-20) against Fas detected Fas at about 45 kDa, and pre-treatment with neuraminidase resulted in a reduction of the molecular weight of Fas of about 8 kDa. These data suggest that the enhancement of Fas-induced apoptosis by pre-treatment with neuraminidase was mediated by a caspase-9 dependent pathway closely associated with the loss of Deltapsim, not by activation of caspase-8, -6 or acidic and neutral sphingomyelinases, and that sialic acid linked to glycoconjugates of Fas may regulate Fas-induced apoptosis in human T cell lymphoma.