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Biomedical subjects

Y X Liu

Publications and source records attributed to Y X Liu.

At least 127 records · Page 7Linked to original sources

Synergism between granulosa and theca-interstitial cells in estrogen biosynthesis by gonadotropin-treated rat ovaries: studies on the two-cell, two-gonadotropin hypothesis using steroid antisera.

In mammalian ovaries, luteinizing hormone (LH) induces androgen biosynthesis by theca interna cells, whereas both follicle-stimulating hormone (FSH) and LH stimulate aromatase activity by granulosa cells. The joint action of these two types of cell and pituitary hormones forms the basis of a 2-cell, 2-gonadotropin hypothesis for biosynthesis of estrogen. We tested the synergism between these cell types using antisera against the estrogen precursors progesterone and testosterone. Ovaries were obtained from immature rats two days after a single injection of pregnant mare serum gonadotropin (PMSG). Granulosa cells were obtained by puncturing the follicles, and ovarian remnants were dissociated with collagenase to derive cells from the theca interstitium. Granulosa and theca interstitial cells were cultured alone or in combination for 20 h. Granulosa cells secreted negligible amounts of estrogen and testosterone, but contained high aromatase activity. Treatment with both FSH and human chorionic gonadotropin (hCG) increased production of progesterone in granulosa cells. In contrast, theca interstitial cells had negligible aromatase activity. Treatment with hCG, but not FSH, increased androgen production by theca interstitium; the hCG action was further augmented by the inclusion of exogenous progesterone. Furthermore, co-culture of gonadotropin-treated granulosa and theca interstitial cells resulted in substantial increases in estrogen production, indicating synergism between the two types of cell. The increases in estrogen production in the co-cultures were accompanied by decreases in progesterone content. To test the possibility that progesterone released by granulosa cells may serve as a substrate for production of androgen in theca cells, specific antiserum was used to adsorb progesterone present in the medium. Addition of the progesterone antibody inhibited gonadotropin-stimulated production of testosterone (39% decrease) and estrogen (64% decrease) by the combined cell cultures. The inhibitory effect on estrogen production could be reversed by the addition of progesterone (10(-6) M) or testosterone (10(-6) M) but not by the addition of a synthetic progestin, R5020. Since testosterone released from theca cells may serve as the substrate for aromatases in granulosa cells, specific testosterone antiserum was also used. Production of estrogens by the combined cell cultures was inhibited (78%) by the testosterone antiserum, but the inhibitory effect was completely reversed by exogenous testosterone or progesterone. Thus, synergistic interactions between granulosa and theca interstitial cells are important in effecting maximal estrogen biosynthesis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Identification and regulation of tissue plasminogen activator activity in rat cumulus-oocyte complexes.

Plasminogen activators convert plasminogen into plasmin, a serine protease that initiates extracellular proteolysis. Two types of plasminogen activator activities have recently been demonstrated in granulosa cells, and the proteolysis-inducing enzymes are believed to be involved in ovulation. However, little attention has been paid to the presence of these enzymes in oocytes. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by a fibrin overlay technique, we studied plasminogen activator activity in oocytes. Denuded oocytes collected from ovaries of hypophysectomized, estrogen-treated immature rats contained a tissue-type plasminogen activator (tPA), but not urokinase (uPA). In contrast, oocyte-free granulosa cells in these preantral follicles contained uPA, but not tPA. The tPA activity found in oocytes was plasminogen-dependent; incubation with increasing numbers (25-200) of denuded oocytes resulted in a dose-dependent increase in fibrinolysis only in the presence of plasminogen. Cellular localization of tPA was studied in the preantral follicles using an immuno-cytochemical method. Positive tPA staining was detected in the cytoplasm, but not in the germinal vesicle or zona pellucida of the oocytes. Furthermore, analysis using a reverse fibrin-overlay method did not reveal the presence of a plasminogen activator inhibitor. Culturing of denuded oocytes for 24 h increased the cellular content of tPA, but the enzyme activity was not further enhanced by treatment with FSH or forskolin. Also, no tPA activity was detected in the medium. We further studied plasminogen activator activities in the cumulus-oocyte complexes. Although only tPA activity was detected in freshly obtained cumulus-oocyte complexes, incubation for 24 h increased both tPA and uPA activity. Furthermore, tPA, but not uPA, activity was stimulated by treatment with FSH or forskolin. This was accompanied by the secretion of tPA into the medium. The identity of tPA and uPA in the cumulus-oocyte complexes was further confirmed by immunoprecipitation with specific antibodies. Isolation of denuded oocytes and cumulus cells after hormonal stimulation of the cumulus-oocyte complexes suggested that tPA activity was stimulated in both cell types and that the cumulus cells may mediate the action of FSH and forskolin on oocytes. In conclusion, the detection and regulation of tPA activity in cumulus-oocyte complexes suggest possible involvement of this enzyme in ovulation or the process of cumulus cell expansion and dispersion. Changes in oocyte tPA content may also serve as an indicator of oocyte development.

Animals↗

Anti-idiotypic antibodies raised against anti-metenkephalin antibodies in rabbits.

Anti-idiotypic antibodies were raised in rabbits immunized against immunoglobulins purified from anti-metenkephalin antibody. The antibodies competed dose-dependently with metenkephalin in binding to anti-metenkephalin antibody on a solid phase metenkephalin enzyme-linked immunosorbent assay (ELISA). The titers of the antiidiotypic activity appeared characteristically in transient peaks and troughs in alternation with anti-metenkephalin activities and were retained after purification on various affinity columns.

Animals↗

[Interaction of human embryo cells derived from different tissues with lymphoblastoid cell lines].

Cells from skin and muscle of two month human embryo and from nasopharynx, tonsil, lung and kidney of four month human embryo were cocultivated with lymphoblastoid cells--B95-8 and Raji. It is found that all the epithelial cells, such as the focal growth of epithelioid cells from the tonsil and nasopharynx as well as kidney cells from the human embryo can not adhere to any kind of lymphoblastoid cells; but fibroblasts from muscle and skin of two month embryo or from nasopharynx, tonsil and lung of four month embryo have the ability of sticking to lymphoblastoid cells. However, only the fibroblastoid cells from the nasopharynx of human embryo have the ability to stick and to fuse with lymphoblastoid cells--B95-8 in cocultivation and to activate the EBV production. The mechanism of this phenomenon and its biological functions will be studied in detail.

Cell Adhesion↗

[Fetal liver cell transfusion in chemotherapy of malignant tumors and blood diseases].

From Feb. to Dec. 1985, 15 patients were treated with fetal liver cell transfusion (FLT). They were 10 cases with malignant tumors treated by chemotherapy (tumor group) and 5 with blood diseases (blood group). Under strict aseptic technique, 3 1/2-6 month old fetus was selected for preparing suspension of the fetal liver cells on the superclean table. The speed of FLT should be increased gradually from slow to rapid. There are 1.8 X 10(8)-4 X 10(12) fetal liver cells in a fetus of more than 5 months old, in which most are CFU-C. In the tumor group, after FLT, white blood cell, platelet and hemoglobin increased by 700-3,900/mm3, 5,000-116,000/mm3 and 0.5-2.0 gm/mm3 respectively. But in the blood group, they increased by 800-1,300/mm3, 14,000-84,000/mm3 and 0.4-11.0 gm/mm3. In most of the cases, these hematological indexes reached up to the highest in 2 weeks after FLT. It suggests that FLT can improve the peripheral blood picture obviously and stimulate bone marrow. Experiment confirms that there are considerable hematopoietic stem cells in the fetal liver, by which the functions of hematopoiesis and immunity are able to recover. It is more marked in the tumor group than in the blood group. FLT provides a favourable condition for high dose chemotherapy.

Agranulocytosis↗

Selenium intake and metabolic balance of 10 men from a low selenium area of China.

Selenium intake and urinary and fecal Se excretion of 10 healthy men from a low Se area in China were determined for three consecutive days, in summer, fall, and winter of 1983, and the spring of 1984 while self-selected diets were being consumed. Mean daily Se intake was 8.8 micrograms/day with a range of 2.3-35.5 micrograms/day, and was far below the recommended range of safe and adequate Se intake of 50-200 micrograms Se/day (National Academy of Sciences/National Research Council). Mean urinary and fecal Se outputs were 3.7 and 3.4 micrograms Se/day, respectively. Mean Se balance during this time was +1.8 micrograms Se/day. Apparent absorption of Se approximated 57%. The low Se intake in this area is a cause for concern since the residents of Molimo may be at risk for Se deficiency diseases.

Adult↗

Bioavailability of selenium to residents in a low-selenium area of China.

For 8 wk 5 groups of 10 men each were given 0.5 g/day DL-methionine, 150 micrograms Se/day as sodium selenite with or without methionine or 150 micrograms Se/day as selenomethionine with or without methionine. Twenty subjects received placebo as controls. Initially plasma Se rose more rapidly than RBC Se. Increases in Se levels were significantly greater with selenomethionine than with the selenite supplement. In the placebo and methionine supplemented groups neither plasma nor RBC Se varied significantly over the course of the study. Supplementation with selenium resulted in marked increases in plasma and RBC GSH-Px within 2 and 4 wk, respectively. Plasma and RBC GSH-Px activity did not differ significantly between Se-supplemented groups. These studies suggest that selenomethionine-Se was more effective in raising plasma and RBC Se than was selenite-Se. Methionine supplements may increase the bioavailability of selenium in severely deficient subjects.

Adult↗

Autocrine role of estrogens in the augmentation of luteinizing hormone receptor formation in cultured rat granulosa cells.

The effects of estrogens on gonadotropin-stimulated luteinizing hormone (LH) receptor formation were examined in primary cultures of rat granulosa cells. Granulosa cells were cultured for 3 days with increasing concentrations of follicle-stimulating hormone (FSH) in the presence or absence of native and synthetic estrogens. Follicle-stimulating hormone stimulated LH receptor formation in a dose-dependent fashion, and estrogens enhanced the FSH-stimulated LH receptor content by decreasing the apparent ED50 of FSH. At 6.25 ng/ml FSH, the enhancement in LH receptor was estrogen dose dependent, with an ED50 value of about 3 X 10(-9) M for 17 beta-estradiol. The increased LH receptor content seen in cells treated with FSH and estrogen was correlated with increased cAMP production by these cells in response to LH stimulation. Time course studies revealed enhancement of FSH-stimulated LH receptor induction at 48 and 72 h of culture. Granulosa cells were also cultured with FSH for 2 days to induce functional LH receptors, then further cultured for 3 days with LH in the presence or absence of estrogens. At 30 ng/ml LH, increasing concentrations of estrogens maintained LH receptor content in a dose-dependent fashion, with their relative estrogenic potencies in keeping with reported binding affinities to estrogen receptors. An autocrine role of estrogens on LH receptor formation was further tested in granulosa cells treated with FSH and an aromatase substrate (androstenedione) to increase estrogen biosynthesis. Cotreatment with semipurified estrogen antibodies partially blocked the FSH stimulation of LH receptors, whereas nonimmune serum was ineffective. Also, inclusion of diethylstilbestrol prevented the inhibitory effect of the estrogen antibodies. Thus, local estrogens in ovarian follicles may play an autocrine role in granulosa cells to enhance LH receptor formation and to increase granulosa cell responsiveness to the LH surge, with subsequent ovulation and adequate corpus luteum formation.

Animals↗

Isolation and characterization of a novel nucleic acid binding protein from calf lenses.

The transitional process of lens cellular differentiation is accompanied by several unique morphological and biochemical changes. Pyknosis or apoptosis of the nucleus involves extensive degradation of genetic materials. In an attempt to search for a gene product responsible for such a regulatory process, we have adopted DNA-cellulose affinity chromatography to enrich the specific binding protein. A binding protein was isolated by high salt (0.8M KCl) wash of the lens polysomal fraction and purified to apparent homogeneity by DNA-cellulose affinity column and chromatofocusing. The nucleic acid binding protein has an apparent molecular weight of 36,000, designated as regulatory factor 36 (RF-36), as determined by SDS/PAGE. Amino acid composition analysis indicated that RF-36 contains high proportions of glycine, alanine, characteristic of the core heteronucleus RNP proteins. Comparative immunological studies with other DNA binding protein antigen (e.g. helix destabilizing protein) suggest the existence of some common overlapping determinant. However, when monoclonal anti-RF-36 was used as immunoprobe, no cross immunoactivity was detected between these homologous binding proteins, suggesting some antigenic diversity among these two nucleic acid binding proteins from different organisms.

Amino Acids↗

Effect of (E)-5-(2-bromovinyl)-2'-deoxyuridine on several parameters of Epstein-Barr virus infection.

The selective and potent anti-herpesvirus drug, (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVdU), has been examined for its inhibitory effects on several parameters of Epstein-Barr virus (EBV) infection in the lymphoblastoid cell lines Raji, P3HR-1, B-95-8 and P3 hybrid cells (a human embryo oropharyngeal cell line fused with a nasopharyngeal carcinoma cell line). At a dosage of 0.03 to 0.1 mM, BVdU caused a marked inhibition of (i) spontaneous viral capsid antigen (VCA) expression in B-95-8 and P3 hybrid cells, (ii) VCA expression and DNA synthesis in B-95-8 cells induced with croton oil and n-butyrate, (iii) early antigen (EA) expression and DNA synthesis in Raji cells superinfected with EBV, and (iv) VCA expression and DNA synthesis in B-95-8 cells superinfected with EBV. In its inhibitory effects on these various parameters of EBV infection, BVdU appears to be comparable to acyclovir [9-(2-hydroxyethoxymethyl)guanine], another selective anti-herpesvirus drug which has been previously recognized as an effective inhibitor of EBV replication.

Acyclovir↗

Follow-up studies on Epstein-Barr virus IgA/VCA antibody-positive persons in Zangwu County, China.

Serological mass surveys were carried out in Zangwu County, China, using an immunoenzymatic test. 3,533 persons were found to have Epstein-Barr virus (EBV) IgA/VCA antibody among 148,029 persons age 30 years and older who were tested during 1978-1980. Among the IgA/VCA antibody-positive persons, 55 nasopharyngeal carcinoma (NPC) cases were detected. Follow-up studies were carried out yearly on the IgA/VCA antibody-positive persons for 1-3 years, and 32 additional NPC patients were diagnosed. IgA/VCA antibody was detected 8-30 months (average, 13 months) prior to the clinical diagnosis of stage I NPC. There was no marked difference in geometric mean titers of IgA/VCA antibody between the period before onset of NPC and after diagnosis at stage I, but antibody titers were higher during stages II-IV. The NPC detection rates for all persons tested serologically and for IgA/VCA antibody-positive persons, respectively, was 2- and 82-fold the annual incidence of NPC in the general population of the same age group. These data further indicate that serological testing is valuable for the diagnosis of NPC, especially in its early stages, and that EBV may play an important role in the development of NPC.

Adult↗