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Background & Aim: Shortage of donor hepatocytes limits hepatocyte transplantation for clinical application. Induced hepatic stem cells (iHepSCs) have capacities of self‐renewal and bipotential differentiations. Here, we investigated whether iHepSCs could be extensively expanded, and whether they could differentiate into sufficient functional hepatocytes as donors for transplantation therapy after their extensive expansions. Methods: Murine extensively expanded iHepSCs (50‐55 passages) were induced to differentiate into iHepSC‐Heps under a chemically defined condition. iHepSC‐Heps were proved for carrying morphological hepatocyte characters and hepatocytic functions including low‐density lipoprotein uptake, glycogen storage, CLF secretion, ICG uptake and release, Alb secretion, urea synthesis and metabolism‐relative gene expressions respectively. Next, both iHepSCs and iHepSC‐Heps were transplanted into Fah‐/‐ mice respectively. Both liver repopulation and alleviation of liver function were compared between two transplantation groups. Results: Murine iHepSCs still maintained the capacities of self‐renewal and bipotential differentiations after extensive expansion. The efficiency for the functional hepatocyte differentiation from extensively expanded iHepSCs reached to 72.64%. Transplantations of both extensively expanded iHepSCs and iHepSC‐Heps resulted in liver engraftment in Fah‐/‐ mice. Survival rate of Fah‐/‐ mice recipients and level of liver repopulation were 50% and 20.32 ± 4.58% respectively in iHepSC‐Heps group, while 33% and 10.4 ± 4.3% in iHepSCs group. Conclusions: Extensively expanded iHepSCs can efficiently differentiate into hepatocytes in chemical defined medium. Transplantation of iHepSC‐Heps was more effective and more efficient than transplantation of iHepSCs in Fah‐/‐ mice. Our results suggested an innovative system to obtain sufficient hepatocytes through hepatic differentiation of iHepSCs generated by lineage reprogramming. [ABSTRACT FROM AUTHOR]

Objective. The aim of this study was to investigate the potential role of IL-10 in regulating the receptivity marker HOXA10 in the endometrium of women with adenomyosis. Methods. The expression levels of IL-10, HOXA-10, STAT3, and p-STAT3 in the endometrium of women with adenomyosis and controls were examined by means of western blotting and immunohistochemistry. The expression of the HOXA10 protein in Ishikawa cells treated with rIL-10 was examined by western blotting. The attachment rate of BeWo cell spheroids to Ishikawa cells treated with rIL-10 was expressed as a percentage of the total number of spheroids. Results. The expression levels of HOXA10 and IL-10 in the adenomyosis group were significantly lower than those in the control group, and there was a positive correlation between HOXA10 and IL-10 protein levels in all the women examined. rIL-10 increased HOXA10 expression in a concentration- and time-dependent manner by inducing the phosphorylation of STAT3 in Ishikawa cells. Treatment with rIL-10 promoted the attachment of BeWo spheroids to Ishikawa cells, which was reversed by the inhibition of STAT3 phosphorylation. The expression of p-STAT3 in the adenomyosis group was significantly lower than that in the control group, and there was a positive correlation between IL-10 and p-STAT3 protein levels in all the women examined. Conclusions. Both IL-10 and HOXA10 levels in the endometrium are significantly reduced in women with adenomyosis compared with those in control women. The phosphorylation of STAT3 has been proven to be a critical mediator between IL-10 and HOXA10, which may play critical roles in embryo implantation. [ABSTRACT FROM AUTHOR]

Polycystic ovary syndrome (PCOS) is a common endocrine, metabolic and heterogeneous disorder in women of reproductive age, the exact etiology of which remains unknown. To unravel the molecular mechanisms underlying the hyperandrogenic phenotype of PCOS, prenatally androgenized (PNA) mice were used to mimic this phenotype in women with PCOS. Using microarray analysis, 1188 differentially expressed genes, including 671 upregulated and 517 downregulated genes, were identified in ovaries from PNA mice. Five differentially expressed genes (Aldh1a7, Bhmt, Mtr, Nrcam, Ptprg) were validated, and decreased MTR expression was shown in ovaries of PNA mice. In addition, results from qRT-PCR showed decreased MTR expression in granulosa cells (GCs) from women with the hyperandrogenic phenotype of PCOS. Serum levels of S-adenosyl methionine (SAM), the downstream product of MTR, were also decreased in PNA mice and women with the hyperandrogenic phenotype of PCOS. Our study provides evidence that the hyperandrogenic phenotype of PCOS is linked to abnormal folate one-carbon metabolism. [ABSTRACT FROM AUTHOR]

Highlights: [•] miR-145 protects against cardiomyocyte hypertrophy induced by the βAR agonist ISO. [•] GATA6 is a direct functional target gene of miR-145. [•] miR-145 attenuates GATA6 nuclear translocation through inhibition of GSK3β phosphorylation. [ABSTRACT FROM AUTHOR]

Activin, a member of the transforming growth factor-β superfamily, promotes the growth of preantral follicles and the proliferation of granulosa cells. However, little is known about the role of microRNAs in activin-mediated granulosa cell proliferation. Here, we reported a dose- and time-dependent suppression of microRNA-181a (miR-181a) expression by activin A in mouse granulosa cells (mGC). Overexpression of miR-181a in mGC suppressed activin receptor IIA (acvr2a) expression by binding to its 3′-untranslated region (3′-UTR), resulting in down-regulation of cyclin D2 and proliferating cell nuclear antigen expression, leading to inhibition of the cellular proliferation, while overexpression of acvr2a attenuated the suppressive effect of miR-181a on mGC proliferation. Consistent with the inhibition of acvr2a expression, miR-181a prevented the phosphorylation of the activin intracellular signal transducer, mothers against decapentaplegic homolog 2 (Smad2), leading to the inactivation of activin signaling pathway. Interestingly, we found that miR-181a expression decreased in ovaries of mice at age of 8, 12, and 21 days, as compared with that in ovaries of 3-day old mice, and its level was reduced in preantral and antral follicles of mice compared with that in primary ones. Moreover, the level of miR-181a in the blood of patients with premature ovarian failure was significantly increased compared with that in normal females. This study identifies an interplay between miR-181a and acvr2a, and reveals an important role of miR-181a in regulating granulosa cell proliferation and ovarian follicle development. [ABSTRACT FROM AUTHOR]

Problem The objective of this study was to determine whether there was any association between the peripheral blood CD4+ CD25+ Foxp3+ regulatory T cells ( Treg cells) and implantation success in patients undergoing in vitro fertilization ( IVF) treatment. Method of study Prospective observational study of 101 randomly selected women who underwent IVF treatment for tubal factor from May 2011 to June 2011. The percentage of peripheral blood Treg cells and the expression levels of Foxp3 and CTLA4 mRNA in peripheral blood mononuclear cells ( PBMCs) were recorded and their relations to IVF treatment outcomes were analyzed. Results Treg cells were significantly elevated in the pregnant group ( P = 0.03). The expression level of Foxp3 mRNA in PBMCs from pregnant group also significantly increased ( P = 0.02). A receiver operating characteristic analysis (area under curve = 0.631) found that those women with Treg cells >0.6%, the pregnancy rate and live birth rate were much higher as compared to women with Treg cells below this level ( P < 0.05). An increase of Treg cells in the peripheral blood was associated with a better IVF treatment outcome ( OR 4.3, 95% CI = 1.76-10.48), with a sensitivity of 64%, specificity of 71%. Conclusion An elevated level of circulating Treg cells was associated with increased rates of pregnancy and live birth in IVF treatment. [ABSTRACT FROM AUTHOR]

Abstract: Allophycocyanin is a phycobiliprotein with various biological and pharmacological properties. An expression vector was constructed using CpeS as the bilin lyase for the allophycocyanin β subunit, resulting in overexpression of a fluorescent allophycocyanin β-subunit in Escherichia coli. A high-density cell culture was developed using a continuous feeding strategy. After 16 h of culture, the dry cell density reached 21.4 g l−1, the expression of the allophycocyanin β-subunit was 0.86 g l−1 broth, and the relative chromoprotein yield was 81.4%. The recombinant protein showed spectral features similar to native allophycocyanin, which provide an efficient methodology for large-scale production of this valuable fluorescent protein. [Copyright &y& Elsevier]

Chemotherapy treatment in women can frequently cause damage to the ovaries, which may lead to primary ovarian insufficiency (POI). In this study, we assessed the preventative effects of hyaluronic acid (HA) in immunosuppressive drug-induced POI-like rat models and investigated the possible mechanisms. We found that HA, which was reduced in primary and immunosuppressant-induced POI patients, could protect the immunosuppressant-induced damage to granulosa cells (GCs) in vitro. Then we found that HA blocked the tripterygium glycosides (TG) induced POI-like presentations in rats, including delayed or irregular estrous cycles, reduced 17 beta-estradiol(E2) concentration, decreased number of follicles, destruction of follicle structure, and damage of reproductive ability. Furthermore, we investigated the mechanisms of HA prevention effects on POI, which was associated with promotion of GC proliferation and PGRMC1 expression. In conclusion, HA prevents chemotherapy-induced ovarian damage by promoting PGRMC1 in GCs. This study may provide a new strategy for prevention and treatment of POI. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to determine whether there was any association between peripheral blood NKT-like cells and IVF treatment outcome. In this prospective study, 124 women who underwent IVF treatment for tubal factor were randomly selected in June and July 2011. Blood samples were obtained on the day of vaginal oocyte retrieval prior to the procedure. The percentages of peripheral blood NKT-like cell subsets and the expression levels of mRNA encoding MCP-1 and TNFα in PBMCs were recorded, and their relation to IVF treatment outcomes was analyzed. CD3+CD56+CD16+ NKT-like cells were significantly elevated in the pregnant group. The expression level of MCP-1 and TNFα mRNAs in PBMCs from the pregnant group was also increased. Furthermore, both MCP-1 and TNFα mRNA expression were positively correlated with the percentage of CD3+CD56+CD16+ NKT-like cells (r =0.55 and r =0.70, respectively). A ROC analysis (AUC=0.634) found that in those women with CD3+CD56+CD16+ NKT-like cells >1.85%, the pregnancy rate and live birth rate were higher than in women with cells below this level. An increase in CD3+CD56+CD16+ NKT-like cells was associated with a better IVF treatment outcome (OR 4.89, 95% CI=1.86–12.88), with a sensitivity of 47%, specificity of 85%, PPV of 89%, and NPV of 56%. We conclude that an elevated level of circulating CD3+CD56+CD16+ NKT-like cells is associated with increased rates of pregnancy and live births in IVF treatment. [Copyright &y& Elsevier]

Abstract: Members of the KLFs family of transcription factors play roles in maternal endometrium development during embryo implantation. However, the specific role of KLF12 in endometrium development has not yet been described. In this study, we showed that KLF12 expression in human endometrial stromal cells (HESCs) was significantly decreased after decidualization stimulated by 8-Br-cAMP and medroxyprogesterone acetate (MPA). The adenovirus-mediated overexpression of KLF12 in HESCs significantly repressed the expression and secretion of decidualization biomarker genes and their products decidual prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP-1) induced by 8-Br-cAMP and MPA. Moreover, CHIP and luciferase reporter assays demonstrated that KLF12 bound to a CAGTGGG element within the decidual prolactin promoter and decreased decidual PRL promoter (dPRL/-2000Luc) activation in a sequence-specific manner. Taken together, these findings suggest KLF12 is a negative regulator of human endometrial stromal cell decidualization. [Copyright &y& Elsevier]