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Biomedical subjects

Sheng Cui

Publications and source records attributed to Sheng Cui.

At least 19 recordsLinked to original sources

Expression of androgen receptor and its co-localization with estrogen receptor-alpha in the developing pituitary gland of sheep fetus.

No information is known concerning the expression of androgen receptor (AR) and its co-localization with estrogen receptor alpha (ERalpha) in the developing pituitary of sheep fetus. In the present study, we detected AR expression and its co-localization with ERalpha in the anterior pituitary of sheep fetus from day 60 of gestation to the postnatal by dual immunochemistry. The results showed that both AR immunoreactivity (AR-ir) and ERalpha immunoreactivity (ERalpha-ir) were predominantly localized in the nuclei of LH positive gonadotropes. The cell counting results showed that the percentage of the anterior pituitary cells expressing AR fluctuated from 13.51 +/- 0.92 to 17.05 +/- 1.83% during the examined stages, but there were no significant differences between sexes and among ages examined (P > 0.05). However, the proportion of AR-ir cells containing LH markedly increased from day 60 of gestation to the neonatal (P < 0.05). The percentage of AR-ir cells expressing ERalpha-ir significantly increased from day 60 of gestation to the neonatal, respectively (P < 0.05), but no significant differences were seen between genders at each stage examined. These results indicate that both AR and ERalpha are mainly expressed in the gonadotropes of anterior pituitary gland of sheep fetuses, whereas the functions and interaction of AR and ERalpha expressions in the developing pituitary gland are required to be elucidated further.

Animals↗

Different quaternary structures of human RECQ1 are associated with its dual enzymatic activity.

RecQ helicases are essential for the maintenance of chromosome stability. In addition to DNA unwinding, some RecQ enzymes have an intrinsic DNA strand annealing activity. The function of this dual enzymatic activity and the mechanism that regulates it is, however, unknown. Here, we describe two quaternary forms of the human RECQ1 helicase, higher-order oligomers consistent with pentamers or hexamers, and smaller oligomers consistent with monomers or dimers. Size exclusion chromatography and transmission electron microscopy show that the equilibrium between the two assembly states is affected by single-stranded DNA (ssDNA) and ATP binding, where ATP or ATPgammaS favors the smaller oligomeric form. Our three-dimensional electron microscopy reconstructions of human RECQ1 reveal a complex cage-like structure of approximately 120 A x 130 A with a central pore. This oligomeric structure is stabilized under conditions in which RECQ1 is proficient in strand annealing. In contrast, competition experiments with the ATPase-deficient K119R and E220Q mutants indicate that RECQ1 monomers, or tight binding dimers, are required for DNA unwinding. Collectively, our findings suggest that higher-order oligomers are associated with DNA strand annealing, and lower-order oligomers with DNA unwinding.

Binding, Competitive↗

Toxic effects of zearalenone and alpha-zearalenol on the regulation of steroidogenesis and testosterone production in mouse Leydig cells.

Zearalenone (ZEA) and its derivative alpha-zearalenol (alpha-ZOL) are produced by fungi of the genus Fusarium and, after ingestion via contaminated cereals, may lead to animal fertility disturbances and other reproductive pathologies. The previous study demonstrated the toxic effects of ZEA and alpha-ZOL through disturbances in male fertility and other reproductive pathologies in mice. In this study, we further examined the direct biological effects of ZEA and alpha-ZOL on steroidogenesis production, primarily in Leydig cells of mice. Mature mouse Leydig cells were purified by Percoll gradient centrifugation and the cell purity was determined by 3beta-hydroxysteroid dehydrogenase (3beta-HSD) staining. To examine ZEA and alpha-ZOL-induced biological consequences, we measured testosterone secretion and transcription level of 3 key steroidogenic enzymes including 3beta-HSD-1, P450scc and StAR, in ZEA and alpha-ZOL/human chorionicgonadotropin (hCG) co-treated cells. Our results showed that ZEA and alpha-ZOL (10(-4) M, 10(-6) M and 10(-8) M) significantly suppressed hCG (10 ng/ml)-induced testosterone secretion. The suppressive effect is correlated with a decrease in the level of transcription of 3beta-HSD-1, P450scc, and StAR (P<0.05).

3-Hydroxysteroid Dehydrogenases↗

Detection and estrogen regulation of leptin receptor expression in rat dorsal root ganglion.

Leptin receptor (OB-R) is a polypeptide consisting of a single transmembrane-spanning component. OB-R widely distributes in various tissues, including the peripheral nervous system (PNS). However, there is no data about the expression of OB-R in the dorsal root ganglion (DRG). In the present study, we first detected the expressions of OB-R protein and mRNA in the rat DRG using the methods of immunohistochemistry, western blot and reverse transcriptase polymerase chain reaction (RT-PCR). Estrogen is known to influence different functions on the DRG. In this study, we observed that 17beta-estradiol (E(2)) can increase the expressions of OB-R protein and mRNA (P<0.05) in ovariectomized rat DRG and these actions can be prevented by tamoxifen which is a specific estrogen receptors (ERs) antagonist. In addition, the results of dual labeling of OB-R with the two ER isoforms, ERalpha and ERbeta indicate that 100% colocalization of ERalpha with OB-R and about 15% colocalization of ERbeta with OB-R in DRG neurons. These results indicate that OB-R is expressed in the rat DRG and E(2) may up-regulate the expression of the OB-R protein and mRNA in an ERalpha-dependent way.

Animals↗

LIM homeodomain proteins Islet-1 and Lim-3 expressions in the developing pineal gland of chick embryo by immunohistochemistry.

LIM homeodomain proteins Islet-1 and Lim-3 expression and their role in nervous tissue and endocrine glands have been reported; however, nothing is known concerning Islet-1 and Lim-3 expression in the developing pineal gland of the chick embryo. The aim of the present study was to determine the ontogeny of Islet-1 and Lim-3 expression in the developing pineal gland of chick embryo using immunohistochemistry. The results showed that Islet-1 and Lim-3 immunopositive cells were first detected in the pineal evagination of chick embryos at day 4 (E4) and E4.5 of incubation, respectively. In the later developing stages, both Islet-1 and Lim-3 immunopositive cells were consistently detected in the follicular and parafollicular pinealocytes throughout the pineal gland. The relative percentage of Islet-1 immunopositive (Islet-1+) cells relative to the total cells was about 6% at E4.5, and then kept increasing (P < 0.05) and reached about 40% by E12.5; this was followed by no obvious changes until the chicks were newly hatched. The change in Lim-3 immunopositive (Lim-3+) cell number was parallel to that of Islet-1, although Lim-3+ cell were significantly fewer than Islet-1+ cell numbers from E4.5 to E8.5 (P < 0.05). Dual immunohistochemical staining results showed that almost all the Lim-3+ cells expressed Islet-1 at every stage examined, and about 90% of Islet-1+ cells were proliferating cell nuclear antigen negative. These results suggest that both Islet-1 and Lim-3 may be involved in regulating the development and functional maturation of the pineal gland, although further studies are required in elucidating the functional roles of Islet-1 and Lim-3 and the related mechanisms.

Animals↗

A new method for translating the asymmetric Ni/Cr-mediated coupling reactions from stoichiometric to catalytic.

[reaction: see text] A new method has been developed for effectively translating the degree of asymmetric induction and the chemical yield achieved in the stoichiometric asymmetric Ni/Cr-mediated coupling to a catalytic asymmetric process via a chiral sulfonamide ligand. It has also been shown that the Ni catalyst plays a central role. Among a number of the Ni catalysts, the 2,9-dimethylphenanthroline/NiCl(2) complex (7) has been found to be the most effective.

Benzene Derivatives↗

Surprisingly efficient catalytic Cr-mediated coupling reactions.

[reaction: see text] With use of 1 mol % of Cr catalyst 5, surprisingly efficient Cr-mediated couplings of aldehydes with various types of nucleophiles have been realized. The catalyst set of Cr catalyst 5 and Ni catalyst 4 is used for alkenylation, alkynylation, and arylation, whereas the catalyst set of Cr catalyst 5 and CoPc (cobalt phthalocyanine) is used for 2-haloallylation, alkylation, and propargylation. Only the Cr catalyst 5 is required for allylation. The reaction rates in DME and THF have been found significantly faster than that in MeCN.

Aldehydes↗

Developmental changes of Islet-1 and its co-localization with pituitary hormones in the pituitary gland of chick embryo by immunohistochemistry.

Although Islet-1 expression in the pituitary gland of early mouse embryo has been previously described, there are no reports concerning the correlation of Islet-1 expression with lineage restrictions in cell types at the later stages of pituitary development. The role of Islet-1 in chickens is also unknown. The purpose of this study was to follow, by using immunohistochemistry, the ontogeny of pituitary Islet-1 and the various cell types that contain Islet-1 throughout chick embryo development. A few Islet-1-immunopositive (Islet-1(+)) cells were first detected in the pituitary primordium in two out of six embryos at embryonic day 5.5 (E5.5), most of the Islet-1(+) cells being ventrally located. As development progressed, many more Islet-1(+) cells were observed throughout the pars distalis. The relative percentage of Islet-1(+) cells amongst the total Rathke's pouch cells was 4.4% at E6.5. This increased significantly, reaching 11.1% by E10.5, followed by no significant change until hatching. Dual immunohistochemistry showed that adrenocorticotrophs, somatotrophs and lactotrophs did not express Islet-1. The cellular types expressing Islet-1 included luteinizing-hormone-positive (LH(+)) gonadotrophs and thyroid-stimulating-hormone-positive (TSH(+)) thyrotrophs. The cells co-expressing LH and Islet-1 were initially detected at E6.5, the proportion of LH(+) cells possessing Islet-1 being about 4%; this increased to 63% at E14.5, followed by no significant changes until hatching. TSH and Islet-1 co-localized cells were first observed at E10.5, with about 37% TSH(+) cell expressing Islet-1; this increased to about 50% by E16.5, after which there was no evident change until hatching. These results suggest that Islet-1 is involved in determining the cell lineages, proliferation, differentiation and maintenance of hormone-secreting functions of pituitary gonadotrophs and thyrotrophs of chick embryo.

Animals↗

Expression of cysteine sulfinate decarboxylase (CSD) in male reproductive organs of mice.

Cysteine sulfinate decarboxylase (CSD) is the rate-limiting biosynthetic enzyme of taurine, but it is still controversial whether the male reproductive organs have the function to synthesize taurine through CSD pathway. The present study was thus undertaken to detect CSD expression in male mouse reproductive organs by RT-PCR, Western blot and immunohistochemistry. The results show that CSD is expressed both at the mRNA and protein levels in the testis, epididymis and ductus deferens. The relative levels of both CSD mRNA and protein increase from the testis to the epididymis and to the ductus deferens. Immunohistochemical results demonstrate that the main cell types containing CSD are Leydig cells of testis, epithelial cells and some stromal cells throughout the efferent ducts, epididymis and ductus deferens. These results suggest that male genital organs have the function to produce taurine through the CSD pathway, although quantifying the relation of CSD expression to taurine synthesis and the exact functions of taurine in male genital organs still need to be elucidated in future studies.

Animals↗

Biochemical analysis of the DNA unwinding and strand annealing activities catalyzed by human RECQ1.

RecQ helicases play an important role in preserving genomic integrity, and their cellular roles in DNA repair, recombination, and replication have been of considerable interest. Of the five human RecQ helicases identified, three are associated with genetic disorders characterized by an elevated incidence of cancer or premature aging: Werner syndrome, Bloom syndrome, and Rothmund-Thomson syndrome. Although the biochemical properties and protein interactions of the WRN and BLM helicases defective in Werner syndrome and Bloom syndrome, respectively, have been extensively investigated, less information is available concerning the functions of the other human RecQ helicases. We have focused our attention on human RECQ1, a DNA helicase whose cellular functions remain largely uncharacterized. In this work, we have characterized the DNA substrate specificity and optimal cofactor requirements for efficient RECQ1-catalyzed DNA unwinding and determined that RECQ1 has certain properties that are distinct from those of other RecQ helicases. RECQ1 stably bound to a variety of DNA structures, enabling it to unwind a diverse set of DNA substrates. In addition to its DNA binding and helicase activities, RECQ1 catalyzed efficient strand annealing between complementary single-stranded DNA molecules. The ability of RECQ1 to promote strand annealing was modulated by ATP binding, which induced a conformational change in the protein. The enzymatic properties of the RECQ1 helicase and strand annealing activities are discussed in the context of proposed cellular DNA metabolic pathways that are important in the maintenance of genomic stability.

Adenosine Triphosphatases↗

RECQ1 helicase interacts with human mismatch repair factors that regulate genetic recombination.

Understanding the molecular and cellular functions of RecQ helicases has attracted considerable interest since several human diseases characterized by premature aging and/or cancer have been genetically linked to mutations in genes of the RecQ family. Although a human disease has not yet been genetically linked to a mutation in RECQ1, the prominent roles of RecQ helicases in the maintenance of genome stability suggest that RECQ1 helicase is likely to be important in vivo. To acquire a better understanding of RECQ1 cellular and molecular functions, we have investigated its protein interactions. Using a co-immunoprecipitation approach, we have identified several DNA repair factors that are associated with RECQ1 in vivo. Direct physical interaction of these repair factors with RECQ1 was confirmed with purified recombinant proteins. Importantly, RECQ1 stimulates the incision activity of human exonuclease 1 and the mismatch repair recognition complex MSH2/6 stimulates RECQ1 helicase activity. These protein interactions suggest a role of RECQ1 in a pathway involving mismatch repair factors. Regulation of genetic recombination, a proposed role for RecQ helicases, is supported by the identified RECQ1 protein interactions and is discussed.

Adenosine Triphosphatases↗

Ontogeny of estrogen receptor (ER) alpha and its co-localization with pituitary hormones in the pituitary gland of chick embryos.

Estrogen is involved in regulating the development and hormone secretion of the anterior pituitary gland following its binding to estrogen receptors (ERs) expressed on pituitary cells. However, the pituitary is comprised of several cell types, and to date, there is no data about the specific cell types expressing ERs in embyonic chick pituitary. We therefore followed, by immunohistochemistry, the ontogeny of the pituitary ER alpha (ERalpha), and the cell types expressing ERalpha throughout chick embryo development. ERalpha immunoreacitivity was restricted to the nuclei of pituitary cells. ERalpha-immunopositive (ERalpha(+)) cells were first detected at embryonic day 6.5 (E6.5), after which ERalpha(+) cells were consistently detected throughout the anterior pituitary gland, although the density of ERalpha(+) cells in the caudal lobe of the pars distalis was higher than that in the cephalic lobe. The proportion of ERalpha(+) cells in the pituitary was about 6% at E8.5; expression increased to 22% by E18.5 of gestation, with no additional change until hatching. Double-labeling of ERalpha and pituitary hormones showed that the dominant cell types expressing ERalpha were gonadotrophs immunopositive for luteinizing hormone (LH); the proportion of ERalpha(+) cells expressing LH increased throughout gestation and reached approximately 57% at hatching. About 2%-6% of thyroid-stimulating-hormone-immunopositive and 1%-2% prolactin-immunopositive cells expressed ERalpha at later stages of embryonic development, but no growth-hormone-positive or adrenocorticotropic-hormone-positive cells expressed ERalpha during the embryonic period. Thus, gonadotrophs are the main cell population expressing ERalpha in the anterior pituitary gland of chick embryo, and ERalpha is involved in regulating the development of the pituitary gland and the maturation of the hormone-secreting function.

Animals↗

Effects of male and female sex steroids on the development of normal and the transient Froriep's dorsal root ganglia of the chick embryo.

Sex steroids can influence developmental processes and support the survival of neurons in the embryonic central nervous system. Recent studies have shown that estrogen receptors are also expressed in the peripheral nervous system, in the dorsal root ganglia (DRG) of chick embryos. However, no studies have examined the effects of sex steroids on development of embryonic DRG. In the present study, 0.2 microg, 1.0 microg, 5.0 microg 10 microg, 20 microg, 25 microg, and 40 microg doses of testosterone or estradiol were delivered to chick embryos at Hamburger and Hamilton stage 18 (E3). The actions of these doses of sex steroids on the development of the C5DRG (fifth cervical ganglion, a "normal" DRG) and C2DRG (a transient ganglion known as a "Froriep's DRG") were then evaluated by quantifying ganglionic volumes, cell number, proliferation, and apoptosis after 1 day of growth to stage 23. We found that both testosterone and estradiol promoted proliferation of cells in both normal DRG and the Froriep's ganglia. By contrast, estradiol significantly increased the number of apoptotic cells, while testosterone strongly inhibited apoptosis. These actions of sex steroids on DRG development were dose-dependent, and C5DRG and C2DRG showed different sensitivities to the applied sex steroids. In addition, the present results demonstrated that specific ER and AR inhibitors (tamoxifen and flutamide) did not influence the effects of 5 microg E2 and 5 microg T on C2 and C5DRG significantly. These results demonstrate that male and female sex steroids can modulate DRG development through an epigenetic mechanism, as had been shown for the central nervous system.

Androgen Antagonists↗

Expression of estrogen receptors in the efferent ductule of male sheep fetuses during gestation.

There is as yet no report about the developmental changes of estrogen receptors (ERs) in the male reproductive system of the sheep fetus. In the present study, the testis, efferent ductule, and epididymis of sheep fetuses were collected at days 70, 90, and 120 of gestation and in the newborn lamb. ER alpha (ERalpha) and ER beta (ERbeta) were detected by immunohistochemistry. The results showed that ERbeta staining was negative in all of the examined tissues throughout gestation, whereas ERalpha immunoreactivity was only located in the nuclei of the efferent ductule epithelium. In addition, both ERalpha staining intensity and the number of ERalpha-positive cells were higher at day 90 of gestation, compared with that at day 70 and at birth. These results suggest that estrogen may play important roles in efferent ductule development in sheep fetuses.

Animals↗

Analysis of the unwinding activity of the dimeric RECQ1 helicase in the presence of human replication protein A.

RecQ helicases are required for the maintenance of genome stability. Characterization of the substrate specificity and identification of the binding partners of the five human RecQ helicases are essential for understanding their function. In the present study, we have developed an efficient baculovirus expression system that allows us to obtain milligram quantities of recombinant RECQ1. Our gel filtration and dynamic light scattering experiments show that RECQ1 has an apparent molecular mass of 158 kDa and a hydrodynamic radius of 5.4 +/- 0.6 nm, suggesting that RECQ1 forms dimers in solution. The oligomeric state of RECQ1 remains unchanged upon binding to a single-stranded (ss)DNA fragment of 50 nt. We show that RECQ1 alone is able to unwind short DNA duplexes (<110 bp), whereas considerably longer substrates (501 bp) can be unwound only in the presence of human replication protein A (hRPA). The same experiments with Escherichia coli SSB show that RECQ1 is specifically stimulated by hRPA. However, hRPA does not affect the ssDNA-dependent ATPase activity of RECQ1. In addition, our far western, ELISA and co-immunoprecipitation experiments demonstrate that RECQ1 physically interacts with the 70 kDa subunit of hRPA and that this interaction is not mediated by DNA.

Adenosine Triphosphatases↗

The nervous supply to the nasal cavity of the Bactrian camel (Camelus bactrianus).

The Bactrian camel is an important domestic animal in some of the desert and semi-desert areas of the world. However, there is no detailed report about the nervous supply to the nasal cavity of the Bactrian camel. In the present study, seven heads of adult Bactrian camels were collected and the nerve distribution in the nasal cavity was dissected grossly. The results demonstrated that the nerves supplying to the nasal cavity included the olfactory nerve, the ethmoidal nerve from the ophthalmic nerve, and the caudal nerve from the maxillary nerve. The general patterns of nervous distribution in the nasal cavity of the Bactrian camel corresponded with those of other domestic animals. However, the terminal nerve was not observed by this gross anatomical method in the Bactrian camel.

Animals↗

The nerve supply to the orbit of the Bactrian camel.

The Bactrian camel is an important domestic animal in some desert and semi-desert areas of the world, but there is no detailed description of the nervous supply to the orbit in this animal. In this study, 10 heads of adult Bactrian camels were collected and the nerves supplying the orbit were examined grossly. The results showed that the nerves supplying the orbit included the optic nerve, oculomotor nerve, trochlear nerve, abducent nerve, the branches of ophthalmic nerve and the maxillary nerve. The anatomical characteristics of the origins, courses and distributions of the nerves to the orbit in the Bactrian camel were described and compared with those of other domestic animals. These results have importance for further research on the comparative neuroanatomy, neurophysiology and the related clinical surgery of the camel.

Animals↗

Parallel changes between the percentage of fetal pituitary cells immunoreactive to oestrogen receptor alpha and the concentration of 17beta-oestradiol in fetal and maternal plasma during gestation in sheep.

The present study was designed to investigate the relationship between fetal sheep pituitary oestrogen receptor (ER) alpha expression and changes in fetal and maternal plasma 17beta-oestradiol (E2) concentrations during gestation. The results revealed that immunoreactivity for ER was located in the nuclei and distributed throughout the fetal pituitary gland during gestation. The percentage of ERalpha-positive cells was approximately 2% of the total cell population in female fetuses at Day 60 of gestation, increased to approximately 7% and 13% of the total cell population at Days 90 and 120 of gestation, respectively, and then declined to approximately 10% at birth. The fetal plasma E2 concentrations were approximately 19 and 71 pg mL(-1) at Days 90 and 120 of gestation, respectively, and decreased to 22 pg mL(-1) after birth. In male fetuses, plasma E2 concentrations and the percentage of ERalpha-positive cells were similar to values in female fetuses throughout gestation, except on Day 120 when the plasma E2 level in female fetuses was significantly higher than in male fetuses. These data demonstrate that changes in the percentage of fetal pituitary ERalpha-positive cells parallel fetal plasma E2 concentrations throughout gestation.

Animals↗