OBJECTIVETo investigate the expression of inhibin B betaB subunits in human testicular tissues.

METHODSEighty-three cases of the azoospermia underwent testicular biopsy. In accordance with the pathological alterations of spermatogenesis, the samples were divided into four groups: Sertoli-cell-only syndrome (n = 21); hypospermatogenesis (n = 20), maturation arrest (n = 24) and almost normal spermatogenesis (n = 18). Immunohistochemical staining for inhibin B betaB subunits was conducted on the paraffin-embedded sections of different spermatogenesis to localize inhibin B betaB subunits in the seminiferous tubules.

RESULTSImmunohistochemically, positive products of inhibin B betaB subunits were found in both the seminiferous tubules and interstitial tissues of the testis as brown or yellow particles in the cytoplasm. Leydig cells and early intermediate spermatogenic cells showed a very strong positivity; Sertoli cells in the seminiferous tubules were mostly positive; peritubular myoid cells showed a weak positive staining; but no positive expression of inhibin B betaB subunits was found in late spermatids and mature sperm.

CONCLUSIONInhibin B may be produced by both Sertoli cells and early spermatogenic cells in the seminiferous tubules.

Adult ; Azoospermia ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Inhibin-beta Subunits ; biosynthesis ; Male ; Testis ; metabolism ; pathology

OBJECTIVETo examine the expression of activin A (ACT A) and transforming growth factor-beta 1 (TGF-beta 1) during mandibular lengthening and elucidate the difference between the role of ACT A and TGF-beta 1 during mandibular distraction osteogenesis.

METHODSkeletally mature white new zealand rabbits were established right mandibular distraction osteogenesis model. The regenerating tissue of animals' lengthened mandibes were harvested at different time points to have immunohistochemistric research of ACT A, TGF-beta 1 protein and analysis ACT A, TGF-beta 1 mRNA by using RT-PCR semiquantitative mean.

RESULTSAT the end of latency period day, positive stain of ACT A were found in the osteoblasts while positive stain of TGF-beta 1 was found in mesenchymal cells. At the end of distraction phase, fibrosis tissue had no stain of ACT A, but had strong stain of TGF-beta 1. At the period of fixation days of 20 days, both cytoplasm of osteoblasts and extracellular matrix in primary mineralization front were strongly stained of ACT A. The osteoblasts, osteoid and osteocytes in peripheral new bone zone were moderately stained of ACT A. TGF-beta 1 had strongly positive stained in fibrosis zone and weekly positive stained in primary mineralization front and peripheral new bone zone. There were also broad activin A stains in cytoplasm of osteoblasts, osteoid and cytoplasm of ACT A, TGF-beta 1 in osteocytes after distraction for 30 days. Activin A mRNA began to express at the end of latency period. Expression for activin A mRNA increased gradually along with the beginning of distraction and at the peak in distraction of 10 days and 20 days, while TGF beta 1 mRNA increased at the peak at the end of latency period.

CONCLUSIONACT A and TGF beta 1 have different role during rabbit mandibular distraction osteogenesis.

Activins ; analysis ; physiology ; Animals ; Female ; Immunohistochemistry ; Inhibin-beta Subunits ; analysis ; physiology ; Mandible ; surgery ; Osteogenesis, Distraction ; Rabbits ; Transforming Growth Factor beta ; analysis ; physiology ; Transforming Growth Factor beta1

Inhibin, which is important for normal gonadal function, acts on the pituitary gonadotropins to suppress folliclestimulating hormone (FSH) secretion. The level and cellular localization of the inhibin isotypes, alpha, beta(A) and beta(B), in the testis of mice were examined during postnatal development in order to determine if inhibin expression is related to testicular maturation. Mouse testes were sampled on postnatal days (PNDs) 1, 3, 6, 18, 48 and 120, and analyzed by Western blotting and immunofluorescence. Western blot analysis showed very low levels of inhibin alpha, beta(A) and beta(B) expression in the testes at days 1 to 6 after birth. The levels then increased gradually from PND 18 to 48-120, and there were significant peaks at PND 48. Inhibin alpha, beta(A) and beta(B) were detected in testicular cells during postnatal development using immunohistochemistry. The immunoreactivity of inhibin alpha was rarely observed in testicular cells during PND 1 to 6, or in the cytoplasmic process of Sertoli cells surrounding the germ cells and interstitial cells during PND 18 to 120. Inhibin beta(A) and beta(B) immunoreactivity was rarely observed in the testis from PND 1 to 6. On the other hand, it was observed in some spermatogonial cells, as well as in the interstitial space between PND 48 and PND 120. We conclude that the expression of inhibin isotypes increases progressively in the testis of mice with increasing postnatal age, suggesting that inhibin is associated with a negative feedback signal for FSH in testicular maturation.
Aging/*physiology ; Animals ; Gene Expression Regulation/*physiology ; Inhibin-beta Subunits/genetics/*metabolism ; Inhibins/genetics/*metabolism ; Male ; Mice ; Mice, Inbred ICR ; Protein Isoforms/metabolism ; Protein Transport/*physiology ; Testis/*metabolism

Carbonic Anhydrase IV ; genetics ; Chloride Channels ; genetics ; Colorectal Neoplasms ; genetics ; Databases, Genetic ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Inhibin-beta Subunits ; genetics

OBJECTIVEThe purposes of our study were to investigate the association between maternal urinary phthalate metabolites and the levels of inhibin B (INHB) and insulin-like factor 3 (INSL3) in the cord blood in a Chinese pregnant population.

METHODSMaternal urine samples in the third trimester of pregnancy of 69 participants were collected and stored, and the samples of cord blood (10 ml) were collected at delivery between June 2011 and September 2012 in a comprehensive hospital of gynecology and obstetrics in Tianjin, China.Four phthalate metabolites, monomethyl phthalate (MMP), monoethyl phthalate (MEP), monobutyl phthalate (MBP), and mono-2-ethylhexyl phthalate (MEHP) were measured in the urine samples using liquid chromatography-tandem mass spectrometry. The levels of INHB, INSL3 in the cord blood were tested by ELISA. Associations of phthalate exposure with INHB and INSL3 levels were determined by spearman correlation and multiple regression model analysis.

RESULTSThe median concentrations of observed metabolites in descending order were 49.74 µg/L for MMP, 24.96 µg/L for MEHP, 19.52 µg/L for MEP and 17.73 µg/L for MBP. The median concentrations of INHB and INSL3 were 89.09 and 106.21 ng/L.Significant negative associations between INHB and MMP(β' = -0.252), MEP(β' = -0.363) or the sum value (∑PAEs) (β' = -0.346) were found by the multiple regression model analysis. For INSL3, only the sum value (β' = -0.313) was inversely significantly associated with the levels of INSL3 in the cord blood.

CONCLUSIONSMaternal urinary phthalate metabolites were associated with INHB and INSL3 in the cord blood in a Chinese population.

Adult ; Diethylhexyl Phthalate ; analogs & derivatives ; urine ; Female ; Fetal Blood ; chemistry ; Humans ; Infant, Newborn ; Inhibin-beta Subunits ; blood ; Insulin ; blood ; Male ; Maternal Exposure ; Phthalic Acids ; urine ; Pregnancy ; Proteins ; Testicular Hormones ; blood ; Young Adult

OBJECTIVETo investigate the effects of the Chinese herbal medicines Fructus Lycii and Radix Astragali on the function of the Sertoli cells in the rat testis and their mechanisms.

METHODSSertoli cells from the testes of the SD rats aged 18 - 22 days were isolated and cultured. The effects of Fructus Lycii, Radix Astragali and the combined administration of the two on the proliferation of Sertoli cells in vitro were detected by MTT assay, and their effects on the level of INHbetaB mRNA transcription in Sertoli cells in vitro were investigated in both normal environment and peroxide-damaging environment by RT-PCR.

RESULTSThe proliferation of Sertoli cells was promoted by either Fructus Lycii or Radix Astragali at high concentration (P < 0.05), and significantly promoted by the combined administration at high concentration (P <0.01). Sertoli cell INHbetaB transcription was significantly up-regulated by Fructus Lycii, Radix Astragali and their combined administration in vitro (P < 0.01). When the level of INHbetaB mRNA in Sertoli cells significantly dropped (P < 0.01) in the presence of injury induced by peroxide (H2O2), it could be elevated by Radix Astragali (P < 0.05) and significantly up-regulated by Fructus Lycii or the combined administration in vitro (P < 0.01).

CONCLUSIONFructus Lycii, Radix Astragali and the combined administration of the two could promote and protect INHbetaB mRNA in Sertoli cells in vitro.

Animals ; Astragalus membranaceus ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; In Vitro Techniques ; Inhibin-beta Subunits ; biosynthesis ; genetics ; Lycium ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects

OBJECTIVETo examine the expression changes of activin beta A, beta C, beta E and follistatin mRNA in the development of rat hepatic fibrosis induced by carbon tetrachloride (CCl(4)).

METHODSHepatic fibrosis was induced in rats by subcutaneous injections of 40% carbon tetrachloride oily solution for a period of 1 to 7 weeks. After carbon tetrachloride injection of 1, 2, 3, 4, 5, 6, and 7 weeks, the 6-12 rats were killed every time. The kinetics of activin beta A, beta C, beta E and follistatin mRNA expression were assessed by the semi-quantity RT-PCR.

RESULTSActivin beta A, beta C, beta E and follistatin mRNA could be detected in normal rat livers. After CCl(4) injection for 2 or 3 weeks, beta A mRNA was transiently decreased and became undetectable, then increased gradually. After CCl injection for 6 and 7 weeks, beta A mRNA level was significantly higher than controls (P<0.01). beta C mRNA could be detected after CCl(4) injection for 1 to 4 weeks and was significantly increased after 5 weeks over controls (P<0.05). beta E mRNA could not be detected after CCl(4) injection for 1 to 5 weeks, but significantly increased after CCl(4) injection for 6 or 7 weeks compared with controls (P<0.01). Except for normal rat liver, no follistatin mRNA was detected in rats after CCl(4) injection.

CONCLUSIONSActivins and follistatin have different expression changes in the development of hepatic fibrosis and the imbalance of activins and follistatin expression may involve in the formation of hepatic fibrosis.

Activins ; genetics ; Animals ; Carbon Tetrachloride ; Follistatin ; Gene Expression ; Inhibin-beta Subunits ; genetics ; Liver Cirrhosis, Experimental ; chemically induced ; genetics ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVE: To determine the effect of soy isoflavones on cell proliferation and the transcription levels of follicle-stimulating hormone receptor (FSHR), inhibin α (INHα), INHβB, androgen binding protein (ABP), transferrin (Tf) and vimentin in testis sertoli cells in SD rats. METHODS: Sertoli cells were cultured in vitro, exposed to daidzein at 0.03, 0.3, 3, and 30 μmol/L and genistein at 0.05, 0.5, 5 and 50 μmol/L, respectively. MTT was used to detect the proliferation of sertoli cells. Real-time PCR was used to detect the relative mRNA expressions of FSHR, INHα, INHβB, ABP, Tf and vimentin. RESULTS: Compared with control groups, cell proliferation and the relative mRNA expression levels of INHβB and ABP in the treated cells showed no significant alternation. The INHα mRNA expression levels were increased in 0.3 and 3 μmol/L Dai and 0.05 μmol/L Gen, while the mRNA expression levels of FSHR were downregulated in 30 μmol/L Dai and Gen at all concentrations. Tf mRNA expression levels were downregulated in 30 μmol/L Dai and 5 μmol/L and 50 μmol/L Gen, and the mRNA expression levels of vimentin were downregulated in 3 and 30 μmol/L Dai and 50 μmol/L Gen. CONCLUSION Soy Isoflavones may have potential detrimental effect on the male reproductive system, as they may impact the function of sertoli cells by downregulating the transcription levels of some important proteins.
Androgen-Binding Protein ; metabolism ; Animals ; Inhibin-beta Subunits ; metabolism ; Inhibins ; metabolism ; Isoflavones ; adverse effects ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, FSH ; metabolism ; Sertoli Cells ; drug effects ; Soybeans ; chemistry ; Testis ; cytology ; drug effects ; Transferrin ; metabolism

OBJECTIVETo investigate the roles of mouse erythroid differentiation and denucleation factor (MEDDF), a novel factor cloned in our laboratory recently, in erythroid terminal differentiation.

METHODSMouse erythroleukemia (MEL) cells were transfected with eukaryotic expression plasmid pcDNA-MEDDF. Then we investigated the changes on characteristics of cell growth by analyzing cells growth rate, mitotic index and colony-forming rate in semi-solid medium. The expressions of c-myc and beta-globin genes were analysed by semi-quantitative RT-PCR.

RESULTSMEL cells transfected with pcDNA-MEDDF showed significant lower growth rate, mitotic index, and colony-forming rate in semi-solid medium (P<0.01). The percentage of benzidine-positive cells was 32.8% after transfection. The expression of beta-globin in cells transfected with pcDNA-MEDDF was 3.43 times higher than that of control (MEL transfected with blank vector, pcDNA3.1), and the expression of c-myc decreased by 66.3%.

CONCLUSIONSMEDDF can induce differentiation of MEL cell and suppress its malignancy.

Activins ; genetics ; pharmacology ; Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Friend murine leukemia virus ; Globins ; biosynthesis ; genetics ; Inhibin-beta Subunits ; genetics ; pharmacology ; Leukemia, Erythroblastic, Acute ; metabolism ; pathology ; Mice ; Proto-Oncogene Proteins c-myc ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; Transfection ; Tumor Cells, Cultured