Ischemia/reperfusion (IR) induces additional damage during the restoration of blood flow to ischemic myocardium. Urolithin B (UB) is one of the gut metabolites of ellagitannins, a class of antioxidant polyphenols, which was found to be protective against oxidative stress in multiple organs. However, the role of UB in cardiovascular disease remains elusive. Adult Sprague Dawley rats were subjected to left anterior descending artery ligation for 30 min followed by 120 min of reperfusion, with or without UB treatment. In vitro, the H9c2 cardiomyocytes were subjected to hypoxia (94 %N-2/5 %CO2/1 %O-2) for 3 h, followed by reoxygenation (74 %N-2/5 %CO2/21 % O-2) for 3 h (HR). UB was found to decrease myocardial infarct size and attenuate the cardiac dysfunction in the rats after IR, and protect against HR injury in H9c2 cardiomyocytes. Mechanistically, UB inhibited autophagy by. activating Akt/mTOR/ULK1 pathway and protected against oxidative stress and caspase 3-dependent cell apoptosis. In particular, UB induced accumulation of p62 and its interaction with Keap1, which promoted Nrf2 nuclear translocation during HR insult. Of note, the protection of UB against superoxide production and apoptotic cell death was compromised with Nrf2 gene silencing. Taken together, our findings suggested that UB protected against myocardial IR injury at least partially via the p62/Keap1/Nrf2 signaling pathway, which highlights the potential of UB as a novel therapy for ischemic heart disease.

Objective To evaluate the effect of mild hypothermia on the expression of mitofusin 2 (Mfn2) during cerebral ischemia-reperfusion (I/R) in rats. Methods Sixty healthy clean-grade male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups (n=20 each) using a random number table method: sham operation group (group S), group I/R and mild hypothermia group (group H). In I/R and H groups, cerebral I/R was induced by inserting a nylon thread with rounded tip into the right internal carotid artery which was occluded for 2 h and then released for reperfusion.In group H, the surface cooling was started immediately after removing the nylon thread, and the rectal temperature was maintained at 32-34 ℃ for 3 h. The neurological deficit were assessed and scored at 24 and 48 h of reperfusion.Animals were then sacrificed, the cerebral cortex in ischemic area was then removed for determination of the viable neuron count (by HE staining), apoptotic neurons (by TUNEL), and expression of Mfn2 (by Western blot). Apoptosis rate was calculated. Results Compared with group S, the neurological deficit score and apoptosis rate were significantly increased, and the count of viable neurons was decreased, the expression of Mfn2 was down-regulated at each time of reperfusion in I/R and H groups (P<0.05). Compared with group I/R, the neurological deficit score and apoptosis rate were significantly deereased, and the count of viable neurons was increased, and the expression of Mfn2 was up-regulated at each time of reperfusion in group H (P<0.05). Conclusion The mechanism by which mild hypothermia attenuates cerebral I/R injury may be associated with up-regulating the expression of Mfn2 in cerebral cortex of rats. Key words: Hypothermia, induced; Reperfusion injury; Brain; Mitochondrial proteins

Abstract Background The intraflagellar transport protein 140 homolog (IFT140) is involved in the process of intraflagellar transport (IFT), a process that is essential for the formation and maintenance of most eukaryotic cilia and flagella. Variants IFT140 have been reported to account for ciliopathy but association with male fertility has never been described in humans. Here we report the identification of two novel variants of IFT140 which caused spermatogenic dysfunction and male infertility. Methods Whole‐exome sequencing was performed in a 27‐year‐old infertile man presented with severe oligozoospermia, asthenozoospermia, and teratozoospermia (OAT) without other physical abnormality. Sanger sequencing was used to verify gene variants in the patient, his healthy brother, and their parents. Morphology and protein expression in the patient's sperm were examined by transmission electron microscopy (TEM) and immunofluorescence staining. Function of gene variants was predicted by online databases. Results Compound heterozygous variants of IFT140: exon16: c.1837G > A: p.Asp613Asn and exon31: c.4247G > A: p.Ser1416Asn were identified in the patient, both of which showed autosomal recessive inheritance in his family, and had extremely low allele frequency in the population. Morphological abnormalities of the head, nucleus, and tails and the absence of IFT140 from the neck and mid‐piece of the patient's spermatozoa were observed. Mutation Taster database predicted a high probability of damage‐causing by both variations. Conclusion This study for the first time reported IFT140 variants that cause infertility in humans.

This study aimed to study the expression of homeobox (HOX)A11-ASI (HOXA11 antisense RNA) long noncoding RNA (IncRNA) and the expression of homeobox A (H0XA9, HOXAI0, HOXA11, and HOXA13) genes in the eutopic (EU) and ectopic (EC) endometria of women with peritoneal endometriosis. A total of 30 women undergoing laparoscopic surgery for peritoneal endometriosis and 15 infertile women without endometriosis were enrolled in this study. Peritoneal EC tissue samples were obtained through surgery. The EU tissues were obtained by curettage. The EC and EU IncRNA and messenger RNA (mRNA) expression levels were measured using real-time reverse transcriptase-polymerase chain reaction. The HOXA11-ASI IncRNA and H0XA9, HOXA10, HOXA11, and H0XAI3 mRNA were expressed at significantly lower levels in the EU than in the EC, that is, in women with peritoneal endometriosis (P <.05). The expression levels of HOXA10 and HOXA11 in the EU were significantly lower in women with peritoneal endometriosis compared to the control group participants (P <.05), whereas the levels of IncRNA (HOXA11-ASI), H0XA9, and HOXA 13 did not differ significantly between the 2 patient groups (P >.05). In conclusion, the study findings suggest that HOXA11-ASI IncRNA may play a role in the development of peritoneal endometriosis, but HOXA11-ASI may not influence endometrial receptivity in endometriosis-associated infertility.

Cell autophagy and cell apoptosis are both observed in the process of hypoxia-induced ischemic cerebral infarction (ICI). Unc-51 like autophagy activating kinase 1 (Ulk1) and FUN14 Domain-containing Protein 1 (FUNDC1) are both involved in the regulation of cell autophagy. This study aimed to investigate the regulatory effects of Ulk1 and FUNDC1 on hypoxia-induced nerve cell autophagy and apoptosis. Cell viability was measured using cell counting kit-8 (CCK-8) assay. Cell apoptosis was detected using Annexin V-PE/7-ADD staining assay. qRT-PCR was used to quantify the mRNA levels of Ulk1 and FUNDC1 in PC-12 cells. Cell transfection was performed to up-regulate the expression of Ulk1. 3-Methyladenine (3-MA) was used as autophagy inhibitor and rapamycin was used as autophagy activator in our experiments. SP600125 was used as c-Jun N-terminal kinase (JNK) inhibitor. Western blotting was performed to analyze the expression levels of key factors that are related to cell autophagy, apoptosis and JNK pathway. We found that hypoxia simultaneously induced apoptosis and autophagy of PC-12 cells. The activation of Ulk1 and FUNDC1 were also found in PC-12 cells after hypoxia induction. Overexpression of Ulk1 promoted the activation of FUNDC1 and prevented PC-12 cells from hypoxia-induced apoptosis. Suppression of Ulk1 had opposite effects. Furthermore, we also found that JNK pathway participated in the effects of Ulk1 overexpression on PC-12 cell apoptosis reduction. To conclude, Ulk1/FUNDC1 played critical regulatory roles in hypoxia-induced nerve cell autophagy and apoptosis. Overexpression of Ulk1 prevented nerve cells from hypoxia-induced apoptosis by promoting cell autophagy.

Oligoasthenoteratozoospermia (OAT) is characterized as low sperm count, decreased sperm motility and structural abnormalities of the sperm head in the same patient. However, very few studies reported the genetic alterations associated with OAT. Here we report a 38-year-old patient with OAT from a consanguineous family, with 2-6 million/mL sperm density, 2.1-3.8% normal sperm morphology and immotile sperm. Whole-exome sequencing (WES) identified homozygous variant c.1259A > G:p.Y420C in the TDRD6 gene. TDRD6 is a testis specific expressed protein that was localized to the chromatoid bodies in germ cells and played an important role in the nonsense-mediated decay pathway. This rare variant co-segregated with the OAT phenotype in this family. Bioinformatic analysis also suggested the variant a pathogenic mutation. Two intracytoplasmic sperm injection (ICSI) cycles were carried out in the patient's wife, but she did not become pregnant after embryo transfer. So the mutations in TDRD6 may be associated with human male infertility and early embryonic lethality.

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women and is characterised by polycystic ovaries, hyperandrogenism and chronic anovulation. Although the clinical and biochemical signs of PCOS are typically heterogeneous, abnormal folliculogenesis is considered a common characteristic of PCOS. Our aim is to identify the altered miRNA and mRNA expression profiles in the cumulus cells of PCOS patients to investigate their molecular function in the aetiology and pathophysiology of PCOS. In this study, the miRNA expression profiles of the cumulus cell samples isolated from five PCOS and five control patients were determined by an miRNA microarray. At the same time, the altered mRNA profiles of the same cumulus cell samples were also identified by a cDNA microarray. From the microarray data, 17 miRNAs and 1263 mRNAs showed significantly different expression in the PCOS cumulus cells. The differentially expressed miRNA-509-3p and its potential target gene (MAP3K8) were identified from the miRNA and mRNA microarrays respectively. The expression of miRNA-509-3p was up-regulated and MAP3K8 was down-regulated in the PCOS cumulus cells. The direct interaction between miRNA-509-3p and MAP3K8 was confirmed by a luciferase activity assay in KGN cells. In addition, miRNA-509-3p mimics or inhibitor transfection tests in KGN cells further confirmed that miRNA-509-3p improved oestradiol (E2) secretion by inhibiting the expression of MAP3K8 These results help to characterise the pathogenesis of anovulation in PCOS, especially the regulation of E2 production.