To study the impact of sperm DNA fragmentation index (DFI) on the assisted reproductive treatment (ART) outcome, including the rate of good quality embryos, clinical pregnancy and miscarriage. Retrospective cohort study We conducted a retrospective analysis of infertile couples undergoing IVF or ICSI treatments at our reproductive center from May 2012 to April 2018. Only infertile couples caused by cervical factors, male factors, pelvic factors, fallopian tube factors or sexual disturbances were included in the study. Inclusive couples are divided into four subgroups according to the age of the female, including the subgroup less than 30 years old, the 30 to 35 year old subgroup, the 35 to 40 year old subgroup and the subgroup more than 40 years old.The sperm DNA fragmentation was evaluated by SCSA assay and flow cytometry. With DFI=30 as the threshold, each subgroup was divided into low DFI group and high DFI group and the ART outcome were respectively compared. 4841 cycles were included, 4117 of which in low DFI group and 724 in high DFI group.The average age of female and male in high DFI group was slightly higher than low DFI group(subgroup less than 30 years old:30.22±31.69 vs 31.69±2.84, P<0.05) in each subgroups.Despite the slight age difference, the baseline factors , including the level of FSH (P>0.05), number of sinus follicles (P>0.05), were not different between low DFI group and high DFI group of four subgroups. There was no statistical difference in oocytes number, 2PN fertilization rate, cleavage rate, number of available embryos(P>0.05) of all 4 subgroups. There were no significant differences in biochemical pregnancy rate(417/676 vs 33/61, P>0.05), clinical pregnancy rate(386/412 vs 32/33, P>0.05), miscarriage rate(48/386 vs 4/32, P>0.05) between low DFI group and high DFI group in four subgroups. The good quality embryo rate of high DFI group were significantly lower than low DFI group in couples received both IVF and ICSI in the subgroup less than 30 years old (IVF: 0.27±0.40 vs 0.16±0.33, P<0.05; ICSI: 0.05±0.20 vs 0.16±0.35, P<0.05), the 30 to 35 year old subgroup (IVF: 0.28±0.41 vs 0.15±0.35, P<0.05; ICSI: 0.05±0.20 vs 0.16±0.34, P<0.05). In the subgroup more than 40 years old, high scores blastocyst rate in high DFI group is significantly lower than in the low DFI group (IVF: 0.18±0.58 vs 0.03±0.18, P<0.05; ICSI: 0.17±0.38 vs 0.00±0.00, P<0.05). This is a well-controlled and large size retrospective study assessing whether DFI affect the ART outcome. By strictly controlled the confounding female factors, we concluded that DFI had adverse impacts on embryo quality in both IVF and ICSI. Interestingly, unlike previous studies, we found that DFI does not affect biochemical pregnancy rate, the clinical pregnancy rate and miscarriage rate after strict control of the woman's factors.

To compare the in vitro fertilisation (IVF) outcomes of poor ovarian responders among women with laparoscopically diagnosed minimal–mild endometriosis (Group A), moderate–severe endometriosis (Group B) and those without endometriosis (Group C). The comparisons were made separately for age groups younger than 35 years and 35 years or older.

Claudin-5, one of the dominant tight junctions (TJs) proteins, plays an important role in maintaining the barrier function in the blood brain and retinal barrier. This study aimed to investigate the effect of claudin-5 overexpression on the interactions of claudin-1 and -2 and barrier functions in primary cultured human retinal pigment epithelium cells (HRPECs) and human retina endothelial cells (HRECs). Lentivirus was used to mediate the overexpression of claudin-5 in retinal cells. Significantly increased mRNA and protein levels of claudin-5 were detected in the transfection group. After the transfected cells grew on the transwell membrane for three weeks, a stable monolayer cell barrier model was established in vitro. The claudins expressions analysis showed that overexpressed claudin-5 significantly increased the expression of claudin-1, while it decreased the expression of claudin-2 in both mRNA and protein level. Co-IP experiments and barrier function assay revealed that claudin-5 overexpression promoted the interactions of claudin-1 and claudin-2 and enhanced the barrier function of retinal cells. Intriguingly, the exogenous expression of claudin-5 induced new interaction pattern between claudin-5 and claudin-1 or -2 in HRPECs, which do not have endogenous claudin-5 expression. In addition, claudin-5 overexpression decreased cell mobility and the sprouting capability of vessel tube formation in vitro. This study demonstrated that claudin-5 has a positive regulation in the formation of retinal barrier. Claudin elements and their interactions can be modulated and that such dynamic properties are important for the functions of TJs, ranging from the regulation of retinal barrier integrity to junction-associated signaling mechanisms.

Using microRNA (miRNA) expression array, we identified that miR-7 was deregulated in colorectal cancer (CRC). We studied the biological role and molecular target of miR-7 in CRC. miR-7 was downregulated in six out of seven colon cancer cell lines. Ectopic expression of miR-7 suppressed colon cancer cell proliferation (P<0.05), induced apoptosis (P<0.05) and caused cell-cycle arrest in G1 phase (P<0.05). The tumor suppressive function of miR-7 was further confirmed in nude mice (P<0.05). The 3′-untranslated region (3′UTR) of Yin Yang 1 (YY1) mRNA contains an evolutionarily conserved miR-7 binding site using in silico searches, luciferase reporter assay and western blot analysis confirmed that miR-7 directly bound to YY1 3′UTR to negatively regulate the protein expression of YY1 in colon cancer cell lines HCT116 and LOVO. Intriguingly, knock-down of YY1 in three colon cancer cell lines (HCT116, LOVO and DLD1) consistently suppressed cell proliferation (P<0.01) and induced apoptosis (P<0.01), indicating the opposite functions of miR-7 and YY1 in CRC. Consistent with these data, ectopic expression of YY1 promoted cell growth by increasing proliferation (P<0.01) and suppressing apoptosis (P<0.001). The tumorigenic ability of YY1 was further confirmed in vivo in xenograft-nude mouse model (P<0.01). In addition, pathway analyses revealed that the oncogenic effect by YY1 was associated with inhibiting p53 and modulating its downstream effectors p15, caspase cascades and C-Jun, and activating Wnt signaling pathway through activating β-catenin, anti-apoptotic survivin and fibroblast growth factor 4. Furthermore, multivariate analysis revealed that patients with YY1 protein high expression had a significant decrease in overall survival, and Kaplan–Meier survival curves showed that these patients had significantly shorter survival than others (P<0.0001). In conclusion, MiR-7 is a novel miRNA with tumor suppressive function in colon cancer by targeting oncogenic YY1. YY1 promotes colon cancer growth through inhibiting p53 and promoting Wnt signaling pathways and serves as an independent prognostic biomarker for CRC patients.

Aims: KH902 is a fusion protein that can bind vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) through its binding ligand taken from the domains of VEGF receptor 1 and VEGF receptor 2 (VEGFR2). This study was to investigate the effects of intravitreal injection of KH902 on the retinas of streptozotocin-induced diabetic rats. Methods: Two weeks after induction of diabetes, the left eyes of diabetic rats in each group received an intravitreal injection of phosphate-buffered saline (PBS), Avastin or KH902 solution, respectively. Four weeks after intravitreal injection, retinal electrophysiological function and the integrity of inner blood retinal barrier (iBRB) were measured by electroretinogram and Evans blue perfusion. The protein levels of VEGF signal pathway were assayed by western blot. The expression and distribution of claudin-5 and occludin were analysed by double immunofluorescent staining under confocal microscope. The expression of VEGFR2 and PlGF was measured by immunohistochemistry. Results: Four weeks after intravitreal injection, KH902-treated rats had better retinal electrophysiological function, less retinal vessel leakage and lower levels of VEGFR2, PI3K, AKT, p-AKT, p-ERK and p-SRC than PBS or Avastin-treated rats. The distribution of claudin-5 and occludin in the retinal vessels of diabetic rats treated by KH902 was smoother and more uniform than those of diabetic rats treated by PBS or Avastin. The expression of PlGF and VEGFR2 in KH902-treated rats was decreased compared with those in PBS or Avastin-treated rats. Conclusions: KH902 could improve retinal electrophysiological function and inhibit the breakdown of iBRB by inhibiting the expression of VEGFR2, PlGF and PI3K, and the activation of SRC, AKT and ERK.