Cloning of Rabbit Bone Morphogenetic Protein 15 and Its Expression During in vitro Maturation of Rabbit Oocytes

Partial cDNA sequence of rabbit BMP15 was cloned by RT-PCR from rabbit ovaries, showing a similarity of 83%-90% with the BMP15 nucleotide sequences in humans, mice, ovine, sheep, cows and pigs. The expression of BMP15 in rabbit cumulus-oocyte complexs during oocytes in vitro maturation （IVM） was measured by fluorescent quantitative RT-PCR method. BMP 15 was expressed at low levels in immature oocytes and increased to the highest level at 16h of IVM, which coincides with the time of cumulus cell expansion, then declined slowly under IVM cultivation. The expression pattern of BMP 15 suggested that it might be important in cumulus expansion in rabbits.

Transforming growth factor β1 is a differentially expressed candidate protein of congestive heart failure with Qi-deficiency-blood-stasis syndrome

OBJECTIVE: To investigate the role of tongue coating fluid protein in regulation of congestive heart failure(CHF) in Qi-deficiency-blood-stasis syndrome.METHODS: We studied patients with CHF(3 patients with Qi-deficiency-blood-stasis syndrome and 3 without Qi-deficiency-blood-stasis syndrome) to investigate differentially expressed proteins. We also included a control group. A biotin label-based antibody array was used for testing tongue coating fluid samples from patients. Net-work analysis of these differentially expressed proteins was conducted using the STRING database,which can predict the relations between differentially expressed proteins and CHF with Qi-deficiency-blood-stasis syndrome.RESULTS: A total of seven differentially expressed proteins were identified, and among these, transforming growth factor β1(TGF-β1) gets a particular attention for us has drawn specific attention.Network analysis showed a homologous relationship of TGF-β1 with bone morphogenetic protein15, which is associated with myocardial fibrosis.CONCLUSION: Occurrence and development of CHF may result from certain DE-proteins and associated signaling pathways. TGF-β1 protein may be a candidate marker for assessing the risk of CHF in Qideficiency-blood-stasis syndrome.

Effect of Er-Xian decoction(二仙汤) on autoimmune premature ovarian failure in mice

OBJECTIVE:To investigate the therapeutic effect of Er-Xian decoction(二仙汤,EXD)on the autoimmune premature ovarian failure(POF)in mice,and its regulatory mechanisms.METHODS:Female BALB mice were treated with a single intraperitoneal injection of zona pellucida 3(ZP3).One week later,mice received low(5 g/kg),moderate(10 g/kg)and high(20 g/kg)doses of EXD by gastrogavage once daily for 90 d.Premarin(0.03 mg/kg)served as the positive control group.Serum levels of estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)were measured by enzyme-linked immunosorbent assay,lymphocyte subtypes were analyzed,and follicular structure differences were observed by hematoxylin andeosin staining.The main mechanisms of POF was investigated by immunohistochemical analysis.RESULTS:Serum E2 levels in POF model mice were decreased,whereas FSH and LH levels were dramatically increased.Serum levels of LH and FSH were reduced in POF model mice treated with EXD(moderate and high doses)and premarin,while serum level of E2 were increased after POF model mice had been treated with EXD and premarin.The CD3+T,CD4+T,CD4+T/CD8+T ratio of mice in the positive control group and high and medium dose groups of EXD increased(P<0.05),and the number of CD8+T cells decreased significantly(P<0.05)when compared with the model group.Bone morphogenetic protein 15(BMP15)and Akt were repressed in autoimmune POF model mice,whereas high expression was observed in control mice and those treated with EXD(moderate and high doses)and premarin.CONCLUSION:EXD is effective in treating ZP3-induced POF in mice and increased expression of BMP-15,and Akt is represented in the mechanism accounting for this therapeutic effect.

Where are the theca cells from: the mechanism of theca cells derivation and differentiation

Mammalian follicles are composed of oocytes,granulosa cells,and theca cells.Theca cells form in the secondary follicles,maintaining follicular structural integrity and secreting steroid hormones.Two main sources of theca cells exist:Wilms tumor 1 positive(Wt1+)cells native to the ovary and Gli1+mesenchymal cells migrated from the mesonephros.Normal folliculogenesis is a process where oocytes,granulosa cells,and theca cells constantly interact with and support each other through autocrine and paracrine mechanisms.The proliferation and differentiation of theca cells are regulated by oocyte-derived factors,including growth development factor 9 and bone morphogenetic protein 15,and granulosa cell-derived factors,including desert hedgehog,Indian hedgehog,kit ligand,insulin-like growth factor 1,as well as hormones such as insulin and growth hormones.Current research on the origin of theca cells is limited.Identifying the origin of theca cells will help us to systematically elaborate the mechanisms of follicular formation and development.

Effects of Dehydroepiandrosterone on Embryo Quality and Follicular Fluid Markers in Patients with Diminished Ovarian Reserves

Background:To examine the effects of dehydroepiandrosterone(DHEA)on in vitro fertilization(IVF)intracytoplasmic sperm injection(ICSI)and the levels of follicular fluid(FF)markers,namely,anti-Müllerian hormone(AMH),insulin-like growth factor(IGF)-1,bone morphogenetic protein(BMP)-15,and growth differentiation factor(GDF)-9,in patients with diminished ovarian reserves(DORs).Methods:116 patients with DOR were randomized into two groups,DHEA group and control group.Each group contained 58 patients.The DHEA group received 75 mg/d of DHEA for 12 weeks prior to the start of IVF treatment,while the control group entered IVF treatment directly.All patients were treated with the same ovarian stimulation protocol.The primary outcome was high-quality embryo yield.Other IVF parameters,such as the clinical pregnancy rate,embryo survival rate,and intact blastomere rate,were compared between the two groups.FF samples from patients of both groups were collected to measure the levels of AMH,IGF-1,DHEA-sulfate,BMP-15,and GDF-9.Blood was also collected on day 3 of the menstrual cycle to define the baseline hormonal profile and to examine ovarian reserve markers.Results:The high-quality embryo yield was higher in DHEA group than that in control group(P=0.033).AMH and IGF-1 concentrations in FF were significantly higher in DHEA group than that in the control group(2.83±1.14 ng/L vs.1.37±0.55 ng/L,P=0.000;94.02±38.28 ng/L vs.74.03±25.46 ng/L,P=0.004,respectively).The BMP-15 level was also higher in DHEA group(relative expression were 1.80±0.41)than that in control group(relative expression were 0.79±0.16,P<0.0001);however,there was no difference in GDF-9 expression between the two groups(relative expression were 1.29±0.54 and 1.16±0.50 respectively,P>0.05)and in the clinical pregnancy rate between the two groups(13.79%vs.7.27%,respectively,P>0.05).Conclusions:In women with DOR undergoing IVF treatment,pretreatment with DHEA may increase the number of high-quality embryos,which may be due to increased levels of AMH,IGF-1,and BMP-15 in the FF.