vvi-miPEP172b and vvi-miPEP3635b increase cold tolerance of grapevine by regulating the corresponding MIRNA genes.

As a kind of small molecular weight proteins, many peptides have been discovered, including peptides encoded by pri-miRNA (miPEPs). Similar as traditional phytohormone or signaling molecular, these peptides participate in numerous plant growth processes. MicroRNAs (miRNAs) play an important regulatory role in plant stress response. While the roles of miPEPs in response to abiotic stress has not been studied now. In this study, to explore whether miPEPs could contribute to low temperature (4ºC) tolerance of plants, the expression pattern of 23 different vvi-MIRs were analyzed by qRT-PCR in 'Thompson Seedless' (Vitis vinifera) plantlets under cold stress (4ºC) firstly, and vvi-MIR172b and vvi-MIR3635b which showed an elevated expression levels were selected to identify miPEPs. Through transient expression, one small open reading frame (sORF) in each of the two pri-miRNAs could increase the expression of corresponding vvi-MIR, and the amino acid sequences of sORFs were named vvi-miPEP172b and vvi-miPEP3635b, respectively. The synthetic vvi-miPEP172b and vvi-miPEP3635b were applied to the grape plantlets, and the tissue culture plantlets exhibited a higher cold tolerance compared with the control groups. These results revealed the effective roles of miPEPs in plant cold stress resistance for the first time, providing a theoretical basis for the future application of miPEPs to agricultural production.

A miRNA-Encoded Small Peptide, vvi-miPEP171d1, Regulates Adventitious Root Formation.

One of the biggest challenges in clonal propagation of grapevine (Vitis vinifera) is difficulty of rooting. Adventitious root initiation and development are the critical steps in the cutting and layering process of grapevine, but the molecular mechanism of these processes remains unclear. Previous reports have found that microRNA (miRNA)-encoded peptides (miPEPs) can regulate plant root development by increasing the transcription of their corresponding primary miRNA. Here, we report the role of a miPEP in increasing adventitious root formation in grapevine. In this study, we performed a global analysis of miPEPs in grapevine and characterized the function of vvi-miPEP171d1, a functional, small peptide encoded by primary-miR171d. There were three small open reading frames in the 500-bp upstream sequence of pre-miR171d. One of them encoded a small peptide, vvi-miPEP171d1, which could increase the transcription of vvi-MIR171d Exogenous application of vvi-miPEP171d1 to grape tissue culture plantlets promoted adventitious root development by activating the expression of vvi-MIR171d Interestingly, neither exogenous application of the vvi-miPEP171d1 peptide nor overexpression of the vvi-miPEP171d1 coding sequence resulted in phenotypic changes in Arabidopsis (Arabidopsis thaliana). Similarly, application of synthetic ath-miPEP171c, the small peptide encoded by the Arabidopsis ortholog of vvi-MIR171d, inhibited the growth of primary roots and induced the early initiation of lateral and adventitious roots in Arabidopsis, while it had no effect on grape root development. Our findings reveal that miPEP171d1 regulates root development by promoting vvi-MIR171d expression in a species-specific manner, further enriching the theoretical research into miPEPs.

Quantitative histological analysis and ultrastructure of the aging human testis.

PURPOSE: To quantitatively assess the histological and ultrastructural changes resulting from aging in the human testis. METHODS: Age-related histological and ultrastructural changes were evaluated using light microscopy, transmission electron microscopy (TEM) and immunohistochemistry on 41 testicular samples obtained from elderly men and, respectively, assigned to group A (n = 20), 54-69 years old or group B (n = 21), 70-89 years old. Testicular samples derived from 17 young men were used for control. RESULTS: The numbers of Sertoli cells in the aged groups were significantly lower than that in the controls (p < 0.05). With the exception of the Sertoli cell ratios (germ cells/Sertoli cells) of spermatogonia and primary spermatocytes, results showed lower levels of the Sertoli cell ratios of round spermatids and elongated spermatids in the elderly men compared with the young men (p < 0.05). A similar degenerative pattern of the organelles was shown in germ cells and Sertoli cells in the aging testes under TEM. Immunohistochemistry revealed an increased apoptosis index (AI) (0.81 ± 0.13) accompanied by a decreased proliferation index (PI) (30.08 ± 4.86) in the group B (p < 0.05), while both AI and PI were similar between the group A (0.54 ± 0.06; 36.38 ± 7.38) and the controls (0.50 ± 0.15; 40.55 ± 7.92) (p > 0.05). CONCLUSIONS: Aging has negative influence on testicular morphology and spermatogenesis, and the failure of spermatogenic cell development is evident from the spermatid level.

Mifepristone in combination with prostaglandins for termination of 10-16 weeks' gestation: a systematic review.

OBJECTIVES: Medical regimens using mifepristone in combination with prostaglandins have been widely available for women undergoing termination of pregnancy (TOP) at 10-16 weeks' gestation in China. We undertook a systematic review to compare different regimens of mifepristone with prostaglandins for TOP at 10-16 weeks' gestation. METHODS: We searched multiple electronic databases for English and Chinese language reports (1990-2007) including MEDLINE, the Cochrane Library and the Chinese Biomedical Literature Database. Included were trials comparing mifepristone with prostaglandins (misoprostol, gemeprost or carboprost (PG05)) to each other for women at 10-16 weeks' gestation. Outcomes were successful abortion rates, induction-to-expulsion time, blood loss and side effects. Data were processed with RevMan 5 software. RESULTS: Nineteen trials comparing mifepristone with prostaglandin (misoprostol and PG05) were found of which 14 contributed to meta-analyses (4206 women). The quality of reports was poor. The effectiveness of vaginal mifepristone/misoprostol was super than mifepristone/PG05 (RR 1.14, 95%CI 1.05-1.22) as was induction-to-expulsion time, blood loss and side effects. When comparing misoprostol/mifepristone 150mg to misoprostol/mifepristone 200mg, no difference in TOP success rates were found (RR 0.98, 95%CI 0.96-1.01). Misoprostol vaginally compared to orally significantly increased the TOP success rate (RR 1.12, 95%CI 1.01-1.24). Gastrointestinal symptoms and fever occurred more often in misoprostol oral group (RR 1.67, 95%CI 1.46-1.91). CONCLUSIONS: Medical regimens of mifepristone/prostaglandins were effective and safe for TOP at 10-16 weeks' gestation. Misoprostol was super than PG05, and misoprostol vaginally was found to have better effectiveness than misoprostol orally. Further research should evaluate the relative effectiveness of medical methods compared to surgery.

Effects of hydrosalpinx on pinopodes, leukaemia inhibitory factor, integrin beta3 and MUC1 expression in the peri-implantation endometrium.

OBJECTIVES: To compare the expression of pinopodes, LIF, integrin beta(3) and MUC1 in the peri-implantation endometrium of women with and without hydrosalpinx. STUDY DESIGN: A prospective observational study in an assisted reproductive unit in a university teaching hospital, including 20 women with hydrosalpinx and 21 women without hydrosalpinx. Endometrial biopsies were performed on day LH+7 or +8. The proportion and density of pinopodes were assessed by scanning electron microscopy. LIF, integrin beta3 and MUC1 were evaluated with immunohistochemical staining. RESULTS: The proportion and the density of pinopodes were not significantly different between the hydrosalpinx and control groups. The LIF, integrin beta(3), and MUC1 expression were significantly reduced in both glandular epithelial cells and endometrial lumen of the hydrosalpinx group when compared with those of the control group. The expression of integrin beta(3) in stromal cells was also significantly lower in the hydrosalpinx group. CONCLUSIONS: The proportion and the density of pinopodes in the peri-implantation endometrium were not affected by the presence of hydrosalpinx while LIF, integrin beta(3) and MUC1 were significantly reduced in patients with hydrosalpinx.

Ubiquitin carboxyl-terminal hydrolase L1 contributes to the oocyte selective elimination in prepubertal mouse ovaries.

Apoptosis of abnormal oocytes is essential for defective oocyte elimination during prepubertal ovary development, and the ubiquitin system regulates the cell apoptosis via the degradation of specific proteins. Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin system, and the UCH-L1-dependent apoptosis is important for spermatogenesis. In the present study, the change in the number of follicles and the expression of UCH-L1 in oocytes were determined in prepubertal mouse ovaries by immunohistochemical techniques. A significant decrease in the follicular pool was found in prepubertal mouse ovaries during the period of day 21 to day 28 after birth, and accordingly, the UCH-L1 protein expression was increased, to some degree in association with Jun activation domain-binding protein 1 (Jab1) and cyclin-dependent kinase inhibitor p27(Kipl). The increased UCH-L1 protein, together with the corresponding changes of Jab1 was detected in morphologically abnormal oocytes of prepubertal ovaries. Through the immunofluorescent colocalization, UCH-L1 was shown concentrating in abnormal oocytes, and a parallel change in Jab1 was also seen. The affinity analysis confirmed the interaction between UCH-L1 and Jab1 in ovaries. These results suggest that UCH-L1 plays an important role, possibly in association with Jab1 and p27(Kipl), in selective elimination of abnormal oocytes during mouse prepubertal development.

The uterine expression of SEC63 gene is up-regulated at implantation sites in association with the decidualization during the early pregnancy in mice.

BACKGROUND: Sec63 is a key component of the protein translocation machinery in the mammalian endoplasmic reticulum (ER), and involved in the post-translation processing of secretory proteins. The aim of this study was to determine the expression pattern of SEC63 gene in mouse uterus during the early pregnancy. METHODS: Real-time quantitative PCR and Western blot analyses were used to evaluate the alteration in levels of uterine SEC63 gene expression during the peri-implantation period in mice. Further, both in situ hybridization and immunohistochemical analyses were performed to examine the spatial localization of SEC63 gene expression in mouse uterine tissues. The presence of Sec63 protein in human uterine tissue was also detected by immunohistochemical analysis. Statistical analysis was carried out using Tukey test. RESULTS: Uterine SEC63 gene expression was up-regulated and predominantly localized in mouse decidual cells during days 5-8 of pregnancy. More interestingly, Sec63 protein was also detected in human decidua of 10-week pregnancy, whereas was not observed in human endometrial tissues both at proliferative and secretory phases of menstrual cycle. CONCLUSION: The pattern of SEC63 gene expression is consistent with a possible role for SEC63 in decidualization.

Effects of ovarian stimulation on endometrial integrin beta3 and leukemia inhibitory factor expression in the peri-implantation phase.

OBJECTIVE: To investigate the effects of ovarian stimulation on endometrial receptivity in the peri-implantation phase. DESIGN: Comparison of integrin beta3 and leukemia inhibitory factor (LIF) expression in endometrial biopsies from moderate responders and high responders with temporally matched natural cycles. SETTING: Clinical and experimental research. PATIENT(S): Patients with cancelled embryo transfer were evaluated. Moderate responders were defined as E2 5,000-15,000 pmol/L on the day of hCG administration (group B, n = 7). High responders were defined as E2 >15,000 pmol/L on the day of hCG administration (group C, n = 8). Healthy and fertile volunteers were used as natural controls (group A, n = 10). INTERVENTION(S): Endometrial biopsy on the day of LH+7/hCG+7. MAIN OUTCOME MEASURE(S): Immunohistochemistry of integrin beta3 and LIF and imaging analysis. RESULT(S): Endometrial integrin beta3 and LIF expressions in the peri-implantation phase were significantly lower in stimulated cycles (including both moderate and high responders) compared to natural controls. Expression of LIF in glandular epithelium in high responders was lower than that in moderate responders. CONCLUSION(S): These data demonstrated that suggested markers of endometrial receptivity were reduced in stimulated cycles compared with natural cycles, and more so in high response cycles compared with moderate response cycles.

Effects of ovarian high response on implantation and pregnancy outcome during controlled ovarian hyperstimulation (with GnRH agonist and rFSH).

BACKGROUND: The study was aimed at investigating the effects of ovarian high response during controlled ovarian hyperstimulation (COH) on implantation and pregnancy outcome in fresh IVF/ICSI cycles, and subsequent frozen-thawed embryo transfer (FET) cycles. METHODS: An analysis of 1,196 cycles using a long protocol with GnRHa and rFSH was performed. A serum oestrial level (peak E(2)) was obtained on the day of hCG administration, and patients were grouped by peak E(2) percentile distribution into 3 groups. Normal responder was set as cut-off concentrations between percentile (P)25 and P75 (Group A: 1,199-3,047 pg/ml, 595 cycles). Moderate high responders were classified as peak E(2) between P75-P90 (Group B: 3,048-4,127 pg/ml, 180 cycles). For the high response group, the E(2) cut-off concentration was set as P90 and above (Group C: >or=4,128 pg/ml, 119 cycles). Oocyte/embryo parameters and clinical outcomes were compared among the 3 groups in fresh cycles and subsequent FET cycles. RESULTS: Comparisons between groups revealed no difference in the quality of oocyte retrieved and in fertilisation rates. Group C showed decreased trends in implantation and pregnancy rates compared with Group A, but statistical significance was reached only for the difference in implantation rates. Implantation and pregnancy rates in FET cycles were similar among the 3 groups. CONCLUSIONS: High serum estrogen levels were detrimental to implantation, but not to the quality of oocytes, which may be due to an adverse effect on endometrial receptivity in COH cycles.