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J Liu

Publications and source records attributed to J Liu.

At least 757 records · Page 42Linked to original sources
PubMed2000

A checkpoint that monitors cytokinesis in Schizosaccharomyces pombe.

Cell division in Schizosaccharomyces pombe is achieved through the use of a medially positioned actomyosin ring. A division septum is formed centripetally, concomitant with actomyosin ring constriction. Genetic screens have identified mutations in a number of genes that affect actomyosin ring or septum assembly. These cytokinesis-defective mutants, however, undergo multiple S and M phases and die as elongated cells with multiple nuclei. Recently, we have shown that a mutant allele of the S. pombe drc1(+)/cps1(+) gene, which encodes a 1,3-(beta)-glucan synthase subunit, is defective in cytokinesis but displays a novel phenotype. drc1-191/cps1-191 cells are capable of assembling actomyosin rings and completing mitosis, but are incapable of assembling the division septum, causing them to arrest as binucleate cells with a stable actomyosin ring. Each nucleus in arrested cps1-191 cells is able to undergo S phase but these G(2) nuclei are significantly delayed for entry into the M phase. In this study we have investigated the mechanism that causes cps1-191 to block with two G(2) nuclei. We show that the inability of cps1-191 mutants to proceed through multiple mitotic cycles is not related to a defect in cell growth. Rather, the failure to complete some aspect of cytokinesis may prevent the G(2)/M transition of the two interphase-G(2) nuclei. The G(2)/M transition defect of cps1-191 mutants is suppressed by a mutation in the wee1 gene and also by the dominant cdc2 allele cdc2-1w, but not the cdc2-3w allele. Transient depolymerization of all F-actin structures also allowed a significant proportion of the cps1-191 cells to undergo a second round of mitosis. We conclude that an F-actin and Wee1p dependent checkpoint blocks G(2)/M transition until previous cytokinesis is completed.

Actins↗
PubMed2000

Cell cycle-dependent localization of the CDK2-cyclin E complex in Cajal (coiled) bodies.

We have found that CDK2 and cyclin E, but not cyclin A, accumulates within Cajal bodies (CBs) in a cell cycle-dependent fashion. In the absence of cyclin E, CDK2 is not enriched in the CB compartment, suggesting that the translocation of CDK2 to CBs is dependent on cyclin E. CDK2 and cyclin E could be recruited to CBs as a functional complex or CBs may serve as 'docking stations' for CDK2-cyclin E activation by CAKs during the G(1)/S transition. Notably, CDK7-cyclin H-Mat1 complexes are known to accumulate in CBs. Treatment of cells with inhibitors of either CDKs (olomoucine, 200 microM) or RNA polymerase I (actinomycin D, 0.05 microgram/ml), results in a striking reorganization of CDK2 and p80 coilin to the nucleolar periphery. Furthermore, we demonstrate that p80 coilin can be phosphorylated by purified CDK2-cyclin E complexes in vitro. Thus coilin and other CB proteins appear to be downstream targets of CDK2-cyclin E complex-mediated signaling pathways regulating cell cycle progression and controlling aspects of CB function. Possible roles for CDK2 and cyclin E in the well-documented association of CBs, histone gene clusters and RNA 3' end processing factors are discussed.

Animals↗
PubMed2000

Expression of low and high density lipoprotein receptor genes in human adrenals.

Corticosteroids are synthesized from cholesterol which may arise from de novo synthesis or from the uptake of low or high density lipoproteins (LDL or HDL). In the present study, we compared the expression and regulation patterns of LDL receptor and CLA-1 (CD36 and LIMPII Analogous-1, an HDL receptor) genes in adult human adrenocortical tissues to shed more light on the relative contribution of LDL and HDL in human adrenal steroidogenesis. By screening 64 normal and pathological adrenal samples by Northern blotting, we found a positive correlation between LDL receptor and CLA-1 mRNA expression in the adrenal tissues (r=0.547; spearman rank correlation test P<0.01). Adrenal tissues adjacent to Cushing's adenomas contained consistently less LDL receptor and CLA-1 mRNA than normal adrenals (Mann-Whitney P<0.05). In primary cultures of normal adrenal cells, accumulation of both LDL receptor and CLA-1 mRNAs was upregulated by ACTH in a dose- and time-dependent manner, with an earlier induction of LDL receptor than CLA-1 mRNA expression. (Bu)(2)cAMP also increased the levels of these two mRNAs. Addition of LDL, but not HDL, into the culture medium increased cortisol production in untreated adrenocortical cells. Both LDL and HDL enhanced ACTH-induced cortisol production, with the effect of LDL much stronger than that of HDL. Our data show that LDL receptor and CLA-1's expression is ACTH-dependent and occurs in parallel in human adrenal tissues. LDL rather than HDL may be used as the preferential source of cholesterol for steroidogenesis in human adult adrenocortical cells.

Adrenal Glands↗
PubMed2000

Expression of inhibin alpha in adrenocortical tumours reflects the hormonal status of the neoplasm.

Inhibins are gonadal glycoprotein hormones whose main endocrine function is to inhibit pituitary FSH secretion. In addition to testes and ovaries, other steroid-producing organs are sites of inhibin alpha subunit expression. To study the role of inhibins in human adrenal gland, we screened a panel of 150 adrenals (10 normal adrenals, 25 adrenocortical hyperplasias, 65 adrenocortical adenomas, 30 adrenocortical carcinomas and 20 phaeochromocytomas) for inhibin alpha expression. mRNA levels of inhibin alpha subunit were studied in 57 samples and all tissues were stained immunohistochemically with an inhibin alpha subunit-specific antibody. Inhibin alpha mRNA was detected in all adrenocortical tissues. Virilizing adenomas possessed a 10-fold higher median inhibin alpha mRNA expression than did normal adrenals. Bilaterally and nodularly hyperplastic adrenals and other than virilizing adrenocortical tumours had their median inhibin alpha mRNA levels close to those of normal adrenals. Immunohistochemically, inhibin alpha subunit was detectable in all normal and hyperplastic adrenals, as well as in 73% of the adrenocortical tumours. However, the percentage of inhibin alpha-positive cells varied greatly in different tumour types. The median percentage of positive cells was 10 in non-functional and Conn's adenomas, 30 in Cushing's adenomas and 75 in virilizing adenomas. In malignant adrenocortical tumours the median percentage of inhibin alpha-immunopositive cells was 20 in non-functional carcinomas, 30 in Conn's carcinomas, 65 in Cushing's carcinomas and 75 in virilizing carcinomas. All phaeochromocytomas were negative for inhibin alpha subunit both at the mRNA level and immunohistochemically. Our data show that inhibin alpha subunit is highly expressed in both normal and neoplastic androgen-producing adrenocortical cells, with less expression in cortisol-producing and hardly any in aldosterone-producing cells. This suggests a specific role for inhibins in the regulation of adrenal androgen production. We did not find any significant difference in inhibin alpha expression between benign and malignant adrenocortical tumours. Thus inhibin alpha gene does not seem to have a tumour suppressor role in human adrenal cortex.

Adrenal Cortex↗
PubMed2000

Expression of activin A and its receptors in human pheochromocytomas.

Activin A (a homodimer of two activin betaA subunits) has been shown to induce the neuronal differentiation of rat pheochromocytoma PC12 cells. We studied activin A and its receptor gene expression in human pheochromocytomas in vivo and in vitro to clarify the potential involvement of activin A in the pathophysiology of these tumors. We first screened 20 pheochromocytomas and nine normal adrenal tissues for activin betaA mRNA expression. Northern blots hybridized with specific oligonucleotide probes detected weak signals for activin betaA transcripts in pheochromocytomas. Both type I and type II activin receptor (ActR-I, ActR-IB and ActR-II) mRNA expression was also detectable in the pheochromocytoma tissues. In primary cultures of pheochromocytoma cells, expression of activin betaA mRNA was readily detectable by Northern blotting, and secretion of activin A into the conditioned medium was confirmed by an enzyme-linked immunosorbent assay. The expression of activin betaA mRNA and secretion of activin A were induced by (Bu)(2)cAMP after 1 and 3 days of treatment (all P<0.05). A protein kinase inhibitor, staurosporine, inhibited the basal and (Bu)(2)cAMP-induced accumulation of activin betaA mRNA (P<0.05). In addition, induction of chromaffin phenotype by dexamethasone also inhibited the basal and (Bu)(2)cAMP-induced expression of activin A at both mRNA and protein levels (all P<0.05). In contrast, the expression of ActR-I and ActR-IB mRNAs was not affected by these agents in cultured pheochromocytoma cells. In summary, activin betaA subunit and activin receptors are expressed in human pheochromocytomas. Production of activin A in cultured pheochromocytoma cells is induced through the protein kinase A pathway, but reduced during chromaffin differentiation. Therefore, activin A may function as a local neurotrophic factor via an auto/paracrine manner in human pheochromocytomas.

Activin Receptors↗
PubMed2000

Regulation of immunoreactive inhibin A and B secretion in cultured human granulosa-luteal cells by gonadotropins, activin A and insulin-like growth factor type-1 receptor.

Inhibins are gonadal glycoproteins with endocrine effects on pituitary FSH secretion and para/autocrine effects on ovarian and testicular function. The purpose of this study was to investigate the endocrine and para/autocrine regulation of inhibin A and inhibin B secretion in human ovarian granulosa-luteal cells. The cells were obtained from women undergoing in vitro fertilization, and the primary cultures were treated with FSH, LH, human chorionic gonadotropin (hCG), activin A, 8-bromo cyclic AMP (8-BrcAMP), staurosporine (a protein kinase C inhibitor) and an antagonist of IGF action (type-1 IGF receptor antibody alpha IR3). The secretion of inhibins was measured by ELISA assays capable of reliably distinguishing between inhibin A and B. FSH, LH, hCG and 8-BrcAMP increased inhibin A secretion on average up to 180% (P<0.01), 192% (P<0.05), 210% (P<0.01) and 243% (P<0.01) respectively of the control level, while their stimulatory effect on inhibin B secretion was less pronounced (up to 167%, P<0.01; 139%, P<0.05; 127%, P>0.05; 133%, P>0.05 of the controls respectively). alpha IR3 decreased inhibin A and B secretion down to 70% (P<0.01) and 50% (P<0.01) respectively of the control. Staurosporine decreased inhibin B secretion down to 49% (P<0.01) of the control; its effect on inhibin A secretion was not significant. Activin A increased inhibin B secretion up to fourfold of the control (P<0.05) while its effect on inhibin A secretion was insignificant. We conclude that gonadotropins via the protein kinase A signal transduction pathway are the main positive regulators of inhibin A and B secretion in human granulosa-luteal cells. The protein kinase C signal transduction pathway seems to be important especially for inhibin B secretion. Locally produced IGFs are probably important inducers of the production of both forms of inhibin in human ovaries while activins seem to upregulate inhibin B secretion.

Activins↗
PubMed2000

Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs.

Thirty-two crossbred barrows were used to investigate the effects of dietary Ca:total P (tP) ratios in phytase-supplemented diets on the apparent absorption of P and Ca in the small intestine, cecum, and colon. Three Ca:tP ratio treatments (1.5:1, 1.3:1, or 1.0:1) were created by adjusting the amount of ground limestone added to the basal low-P grower (.39% tP including .07% added inorganic P) and finisher (.32% tP without added inorganic P) diets. All low-P ratio diets were supplemented with Natuphos phytase at 500 units/kg. A positive control diet without phytase supplementation contained adequate P and Ca to meet dietary requirements. At 123 kg, the pigs were slaughtered and the contents of ileum, cecum, and colon were collected. Lowering the dietary Ca:tP ratio in the diets containing phytase linearly increased (P < .01) the apparent absorption (% and g/d) of P in the small intestine, but Ca absorption was not affected. Pigs fed the low-P diet with a Ca:tP ratio of 1.0:1 had an apparent absorption (g/d) of P or Ca similar to that of pigs fed the control diet, which was adequate in Ca and P. Averaged across all diets, the apparent absorption of P was highest when measured at the cecum, and the apparent absorption of Ca was highest when measured at the colon. In conclusion, lowering the dietary Ca:tP ratio to 1.0:1 in a low-P diet containing phytase increased the apparent absorption of P in the small intestine. Furthermore, a significant amount of P was absorbed in the cecum.

6-Phytase↗
PubMed2000

Expression of two variants of growth hormone receptor messenger ribonucleic acid in porcine liver.

Transcription of GH receptor (GHR) mRNA is initiated from multiple promoters. Most GHR mRNA arise from GHR Promoter 1 (GHR P1) and GHR P2, which transcribe GHR 1A and GHR 1B mRNA, respectively. Our objective was to characterize the expression of GHR 1A and GHR 1B mRNA in liver of neonatal intact (1 d of age) and castrated (14, 28, and 42 d of age) male pigs (Exp. 1; n = 6 per age group), intact male pigs treated with recombinant porcine ST (rpST) or control (Exp. 2; 21, 42, and 77 d of age; n = 4 pigs per treatment per age), and pregnant gilts treated with rpST (n = 6) or control (n = 7) (Exp. 3). Tissue samples were collected at slaughter for mRNA analyses. The porcine GHR 1A and GHR 1B cDNA were cloned and were homologous to GHR cDNA isolated from other species. Ribonuclease protection assays were used to measure GHR 1A and GHR 1B mRNA. Liver expressed GHR 1A and GHR 1B mRNA, whereas muscle, uterus, and ovary expressed GHR 1B mRNA. The GHR 1A mRNA in the liver of neonatal intact and castrated male pigs (Exp. 1) was expressed at very low levels on d 1, 14, and 28, and two of six pigs expressed a high level of GHR 1A on d 42. The GHR 1B mRNA, however, was detected at all ages (d 1 through 42), and the amount of GHR 1B increased (P<.05) on d 42. The liver of intact male pigs (Exp. 2) expressed GHR 1B mRNA by 21 d, whereas a high level of GHR 1A mRNA was not detected until d 42 (P<.10). Administration of rpST had no effect on expression of GHR 1A or GHR 1B mRNA in pigs younger than 77 d (Exp. 2), but it tended to decrease (P<.10) GHR 1A mRNA but not GHR 1B mRNA in pregnant gilts (Exp. 3). In conclusion, GHR mRNA in porcine liver was composed of at least two variants (GHR 1A and GHR 1B). The GHR 1B mRNA was the major GHR mRNA in pig liver before 77 d of age. The GHR 1A mRNA increased after 42 d of age and tended to undergo specific down-regulation in response to rpST in pregnant gilts.

Animals↗
PubMed2000

p53 and Ki67 in adrenocortical tumors.

The p53 tumor-supressor gene has been reported as the most frequent genetic abnormality seen in human malignancies. Here we studied immunohistochemically the expression of p53 in a large series of adrenocortical tumors. The proliferative activity was assessed by the expression of Ki67. Tumor material consisted of 60 adrenocortical adenomas and 27 adrenocortical carcinomas. A tumor was scored as positive for p53 if more than 10% of the cells showed nuclear staining. All adrenocortical adenomas were negative for p53 and the percentage of Ki67 positive cells was mostly 1-2% but never exceeded 5%. Hormonal activity did not reflect the proliferation index. Adrenocortical carcinomas, however, behaved differently depending on hormonal activity. 10/13 of non-functional , 0/3 Conn's, 3/7 Cushing's and 3/4 virilizing carcinomas were positive for p53. The proliferative activity was also higher in non-fuctional carcinomas compared with hormonally active tumors. Our data show that majority of adrenocortical carcinomas are positive for p53, whereas all adenomas are negative. Hormonal activity of carcinomas reflects both p53 status and proliferation index. Thus, immunohistochemical levels of p53 and Ki67 are higher in hormonally inactive adrenocortical carcinomas.

Adenoma↗
PubMed2000

Expression of vascular endothelial growth factor in human adrenals.

Angiogenesis is an important component in many biological processes and also in pathologic conditions including neoplastic diseases. Vascular endothelial growth factor (VEGF) is a secreted endothelial cell-specific growth factor, which is induced by tissue hypoxia and is angiogenic in vivo. Adrenal gland is a well-vascularized organ, and the roles of VEGF in normal adrenal and in adrenal tumorigenesis is not well characterized. We therefore investigated VEGF mRNA expression in normal human adrenals and in cultured adrenocortical cells. VEGF mRNA was constantly expressed in normal adrenals as well as in cultured adrenocortical cells. The mRNA levels were increased after 24h stimulation with either ACTH or cAMP. The effect of cAMP was dose-dependent. This suggests that ACTH-induced VEGF mRNA expression is mediated via protein kinase A dependent pathway.

Adrenal Glands↗
PubMed2000

Detection of the human immunodeficiency virus regulatory protein tat in CNS tissues.

Neuropathologically, human immunodeficiency virus (HIV) is associated with a range of inflammatory disorders, extensive cortical neuronal loss, and dendritic and synaptic damage. Although the mechanisms resulting in these abnormalities are still unclear, the neurotoxic effects are thought to be due in part to viral products including the tat gene product. We have previously shown that Tat when presented to neurons extracellularly interacts with neuronal cell membranes to cause neuronal excitation and toxicity in fmole amounts. To determine the role of Tat in mediating HIV encephalitis (HIVE), we detected tat mRNA and protein in tissue extracts of nine patients with HIVE and seven patients without HIVE. Despite long autopsy times and significant degradation, tat mRNA was detected in 4/9 patients with HIVE but not in any of the seven patients without dementia. Similarly, the env mRNA was also detected in 5/9 patients with HIVE but not in the patients without HIVE. However, vif mRNA was detected in both groups of patients with (5/9) or without (2/7) HIVE. Using protein extracts from the brains of the same groups of patients we were unable to detect Tat by enzyme linked immunosorbant assay (ELISA) (sensitivity of 2 ng Tat/ml of brain tissue). However, Tat could be detected immunohistochemically and in protein extracts from the brains of rhesus macaques with encephalitis due to a chimeric strain of HIV and simian immunodeficiency virus (SHIV). Our observations support the role of Tat in the neuropathogenesis of HIV and SHIV encephalitis.

AIDS Dementia Complex↗
PubMed2000

All-trans retinoic acid modulates fas expression and enhances chemosensitivity of human medulloblastoma cells.

Retinoic acid (RA) can promote human medulloblastoma cells Med-3 toward differentiation but is not sufficient to induce cell death, suggesting its limited effect on medulloblastomas. On the other hand, the differentiated tumour cells have been supposed to be more sensitive to chemotherapeutic drugs. To elucidate this possibility for medulloblastoma cells, 10 microM/l RA, 1.0 microg/ml cisplatin (CP) and their half-dosage combinations were utilized in this study to treat Med-3 cells and their influences in cell proliferation, morphology and death patterns were evaluated. In parallel, the expressions of Fas and its ligand (FasL) were analyzed by immunocytochemical staining and Western blot hybridization. Anti-Fas antibody was used to incubate the Med-3 cells pretreated by 10 microM/l RA or 1.0 microg/ml CP. It was revealed that RA and CP could inhibit cell growth but rarely induce apoptosis. Combination of half doses each of RA and CP effectively caused most of tumour cells to die of apoptosis within 6 days. FasL molecules in 29 kDa and 37 kDa were detected in Med-3 cells with and without the treatments. The Fas molecule around 30 kDa and located in the cytoplasm was found in the normally cultured cells and the cells treated by CP. An additional 45 kDa Fas band with the appearance of its cell surface labeling was detected in the cells treated by 10 microM/l RA and by 5 microM/l RA + 0.5 microg/ml CP. The anti-Fas antibody could efficiently induce apoptosis only in the cell populations pretreated by RA. Our data thus suggest that RA can enhance the chemosensitivity of human medulloblastoma Med-3 cells presumably via modulating the Fas expression pattern. The RA/CP combined regimen would be a potential therapeutic approach for medulloblastomas.

Antineoplastic Agents↗
PubMed2000

Frequent expression of soluble Fas and Fas ligand in Chinese stomach cancer and its preneoplastic lesions.

To investigate the frequency and pattern of Fas and FasL expression in the gastric mucosa at different stages of gastrocarcinogenesis, the combined examinations of pathology, immunocytochemistry and Western blot hybridisation were performed on the cancer specimens as well as their preneoplastic and non-cancerous counterparts. The frequencies of Fas and FasL expression were found to be 6.3% (1/16) and 62.5% (10/16) in non-cancerous mucosa, 60% (6/10) and 80% (8/10) in atrophic gastritis, 75% (9/12) and 83% (10/12) in intestinal metaplasia, 100% in both dysplasia Grades II (20/20) and Grade III (15/15) and 4 types of gastric carcinomas (74/74). Two forms of FasL protein in 37 kDa and 26 kDa were detected in all FasL+ cases. Soluble Fas (30 kDa) but not the membrane-type (43 kDa) is predominantly expressed in the Fas+ cases. Our data thus suggest a close correlation of soluble Fas with stomach tumour progression P<0.01. The sFas protein, together with the tumor-derived soluble and membrane FasL, may confer on the transforming and transformed gastric epithelial cells an immune advantage enabling escape from endogenous and exogenous suicide signal(s).

Fas Ligand Protein↗
PubMed2000

[Quick RNA extraction from specified morphological region in frozen sections and its RT-PCR application].

OBJECTIVE: To establish a method for quick RNA extraction from specified histological region in frozen sections so as to avoid cross RNA contamination among different types of cells/tissues and to achieve specific RT-PCR amplification. METHODS: Prepare serial frozen sections 5microm in thickness for histological and immunocytochemical examination. Based on the results, make one or two pieces of 20 microm section and select a specified region for quick RNA extraction and RT-PCR analysis. RESULTS: RNA extraction can be finished within an hour and the data of RT-PCR obtained within 6 hours. Besides the high performance, this method can efficiently exclude pseudo positive or pseudo negative results caused by inter-cellular RNA contamination. CONCLUSION: This method is an important complement to the current techniques for RNA extraction and RT-PCR.

Frozen Sections↗
PubMed2000

Flow cytometric DNA analyses of epithelial dysplasia of the esophagus.

OBJECTIVE: To investigate, with flow cytometry, DNA aneuploidy as a marker of early carcinogenesis in dysplastic esophageal lesions. STUDY DESIGN: DNA content of exfoliated cells from 789 cases of esophageal dysplasia (including mild dysplasia, 195 cases; moderate dysplasia, 383 cases; and severe dysplasia, 211 cases) was determined with a FACS 420 flow cytometer. RESULTS: Cellular DNA content was closely related to the severity of dysplasia. The carcinogenesis rate in patients with dysplasia showed that DNA aneuploidy was significantly higher than in patients showing DNA diploidy. CONCLUSION: DNA aneuploidy in dysplastic lesions is a very important early signal of carcinogenesis. Patients with dysplastic lesions showing DNA aneuploidy should be treated and closely followed.

Aneuploidy↗
PubMed2000

[Ultrastructure study of hepatocellular carcinoma treated by diamminedichloroplatinum].

OBJECTIVE: To study the changes of the ultrastructure of hepatocellular carcinoma (HCC) after being treated by diamminedichloroplatinum. METHODS: The changes of morphological feature of HCC treated by diamminedichloroplatinum were observed with transmission electron microscope. RESULTS: The cells lost its specialized cell surface elements such as microvilli and cell-cell junctions. The nucleus chromatin condensed and segregated. The cytoplasm was hollow. The ribosomes on the endoplasmic reticulum reduced or lacked. The glycogen granules reduced. The mitochondria vacuolalized or produced flocculi, and the apoptotic bodies could be found. CONCLUSION: The changes of the ultrastructure of HCC treated by diamminedichloroplatinum correspond with the procedure of programmed cell death. Apoptosis of HCC cells can be induced by diamminedichloroplatinum.

Adult↗