Persimmon leaf extract alleviates chronic social defeat stress-induced depressive-like behaviors by preventing dendritic spine loss via inhibition of serotonin reuptake in mice.

BACKGROUND: Fresh or dried Persimmon leaves (Diospyros kaki Thunb.) exhibit preventive effects on cardiovascular and cerebrovascular diseases. However, their antidepressant effects and underlying mechanisms are unclear. Thus, we investigated mechanisms responsible for Persimmon leaf extract (PLE) activity on chronic social defeat stress (CSDS)-induced depressive-like behaviors in mice. METHODS: CSDS was used as a mouse model of depression. We performed the sucrose preference test (SPT), forced swim test (FST), and tail suspension test (TST) to identify depressive-like behavior. Spine density and dendritic morphology were assessed using Golgi staining. Neurochemicals were quantified by microdialysis, doublecortin by immunofluorescence, and cAMP using an ELISA kit. Finally, the levels of cortical proteins of phosphorylated cAMP-response element binding protein (CREB), brain-derived neurotrophic factor (BDNF), postsynaptic density synapsin-1 and protein 95 (PSD95) were quantified by western blot. 16S rRNA gene sequencing was used to detect fecal microbiota. RESULTS: Treatment of CSDS-subjected mice with PLE (30.0-60.0 mg/kg, i.g.) enhanced sucrose preference, decreased immobility times in the TST and FST but did not affect locomotor activity. Furthermore, persistent social defeat stress decreased dendritic spine density and dendritic length in the brain, as well as decreased PSD95 and synapsin-1 expression. PLE, interestingly, inhibited dendritic spine loss and increased synaptic protein levels. PLE also increased brain levels of 5-HT, cAMP, phosphorylated (p)-CREB, BDNF, PSD95, and synapsin-1 in mice subjected to CSDS. Furthermore, PLE increased their doublecortin-positive cell count in the hippocampal dentate gyrus. CSDS mice represented a distinct fecal microbiota cluster which differed compared with normal C57BL/6J mice, and the phenotype was rescued by PLE. CONCLUSIONS: PLE alleviated CSDS-induced depressive behaviors and spinal damage by suppressing serotonin reuptake and activating the cAMP/CREB/BDNF signaling pathway. Simultaneously, PLE influenced the composition of the fecal microbiota in CSDS-subjected mice.

[Impact of sperm function on the fertilization rate of IVF].

Objective: To analyze sperm function-related risk factors and their value in predicting the fertilization rate (FR) of in vitro fertilization (IVF). METHODS: This retrospective study included 668 cases of IVF performed in the Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology from July 2018 to April 2021, which were divided into a low-FR group (FR ≤ 65%, n = 107) and a high-FR group (FR > 65%, n = 561). We compared the sperm volume, sperm concentration, percentages of progressively motile sperm (PMS) and morphologically normal sperm (MNS), acrosome reaction rate, acrosin activity and DNA fragmentation index (DFI) between the two groups, analyzed the correlation between sperm parameters and FR by Spearman's correlation analysis, and the value of sperm function-related risk factors in predicting the FR of IVF using binary logistic regression analysis and receiver-operator characteristic (ROC) curve. RESULTS: There were statistically significant differences in the infertility type of the female patients (P < 0.05) but not in the sperm volume, sperm concentration, PMS, MNS, and acrosome reaction rate (P > 0.05). The DFI was significantly lower in the high-FR than in the low-FR group (ï¼»14.31 ± 4.46ï¼½% vs ï¼»15.35 ± 5.68ï¼½%, P = 0.034) and the acrosin activity remarkably higher in the former than in the latter group (ï¼»102.11 ± 47.18ï¼½ vs ï¼»91.98 ± 42.61ï¼½ µIU/106, P = 0.039). Spearman's correlation analysis showed that the FR was correlated negatively with DFI (r = 0.090, P = 0.020) and positively with acrosin activity (r = 0.079, P = 0.042). Primary infertility and DFI were found to be unfavorable factors while acrosin activity a favorable factor for the FR of IVF. The areas under the ROC curve (AUC) of DFI and acrosin activity were 0.571 and 0.562, respectively. CONCLUSIONS: DFI and acrosin activity are risk factors and have a predictive value for the fertilization rate of IVF.

Rabdosia serra alleviates dextran sulfate sodium salt-induced colitis in mice through anti-inflammation, regulating Th17/Treg balance, maintaining intestinal barrier integrity, and modulating gut microbiota.

Rabdosia serra (R. serra), an important component of Chinese herbal tea, has traditionally been used to treat hepatitis, jaundice, cholecystitis, and colitis. However, the chemical composition of R. serra and its effect against colitis remain unclear. In this study, the chemical composition of the water extract of R. serra was analyzed using ultra performance liquid chromatography coupled with a hybrid linear ion trap quadrupole-orbitrap mass spectrometer (UPLC-LTQ-Orbitrap-MS). A total of 46 compounds, comprising ent-kaurane diterpenoids, flavonoids, phenolic acids, and steroids, were identified in the water extract of R. serra, and the extract could significantly alleviate dextran sulfate sodium salt-induced colitis by improving colon length, upregulating anti-inflammatory factors, downregulating proinflammatory factors, and restoring the balance of T helper 17/T regulatory cells. R. serra also preserved intestinal barrier function by increasing the level of tight junction proteins (zonula occludens 1 and occludin) in mouse colonic tissue. In addition, R. serra modulated the gut microbiota composition by increasing bacterial richness and diversity, increasing the abundance of beneficial bacteria (Muribaculaceae, Bacteroides, Lactobacillus, and Prevotellaceae_UCG-001), and decreasing the abundance of pathogenic bacteria (Turicibacter, Eubacterium_fissicatena_group, and Eubacterium_xylanophilum_group). Gut microbiota depletion by antibiotics further confirmed that R. serra alleviated colitis in a microbiota-dependent manner. Overall, our findings provide chemical and biological evidence for the potential application of R. serra in the management of colitis.

MicroRNAs secreted by human embryos could be potential biomarkers for clinical outcomes of assisted reproductive technology.

Introduction: MicroRNAs (miRNAs) are important regulators of many biological functions, including embryo implantation and development. Recently, it has been reported that miRNAs in biofluids are predictive for physiological and pathological processes. Objectives: In this study, we aim to investigate whether the miRNAs secreted by human embryos in culture medium can be used as embryonic biomarkers. Methods: The culture media were prospectively collected from embryos of patients at reproductive medicine center with informed consent. A high-throughput miRNA sequencing method was applied to detect the miRNA profiles in the human embryo culture media. After bioinformatics analysis and screening of differentially expressed miRNAs, quantitative real-time polymerase chain reaction (qRT-PCR) assay was subsequently performed to further confirm the sequencing results with mixed samples. Furthermore, we performed droplet digital PCR (ddPCR) to verify the target miRNAs at single sample level. Receiver operating characteristic (ROC) analyses were performed for differentially expressed miRNAs. Results: Compared with embryos with failed pregnancy, the embryos with successful pregnancy secreted different miRNA profiles into the culture media, which were predicted to be involved in multiple biological processes. Validated by droplet digital polymerase chain reaction (ddPCR), the expression of hsa-miR-26b-5p and hsa-miR-21-5p in the culture media of cleavage embryos with successful pregnancy was significantly lower than that of embryos with failed pregnancy. Moreover, the Receiver Operating Characteristic (ROC) curve analysis indicated that hsa-miR-26b-5p and hsa-miR-21-5p could serve as potential biomarkers for reproductive outcomes. Conclusion: Together, our findings highlight the important predictive potential of miRNAs secreted by human embryos in culture media, which is meaningful for non-invasive embryo selection in assisted reproductive technology.

Bone morphogenetic protein 6 expression in cumulus cells is negatively associated with oocyte maturation.

Bone morphogenetic protein 6 (BMP6) is a regulatory peptide secreted by oocytes and granulosa cells that locally regulates folliculogenesis and follicular development. To determine BMP6 location, we studied BMP6 expression in human follicles using immunohistochemistry, real-time polymerase chain reaction (RT-PCR) and western blot analysis. RT-PCR was performed on 354 individual cumulus cell (CC) masses from 48 women to investigate the relationship between BMP6 mRNA expression in CCs and oocyte developmental potential. Results showed that BMP6 protein was mainly located in oocytes from preantral follicles and in granulosa cells from antral follicles. BMP6 mRNA expression was much higher in oocytes than in CCs and mural granulosa cells (mGCs) from preovulatory follicles (p < 0.01), and BMP6 protein level was higher in CCs than in mGCs (p < 0.05). BMP6 mRNA expression was higher in CCs from immature oocytes than in those from mature oocytes (p < 0.05). However, BMP6 mRNA expression in CCs was not associated with oocyte fertilization, embryo morphological grading, or implantation. In conclusion, BMP6 was mainly expressed in oocytes at all human follicular developmental stages and BMP6 mRNA expression in CCs may be negatively correlated with oocyte maturation. BMP6 expression could therefore be used as a biomarker of oocyte maturation.

Cell-free mitochondrial DNA in human follicular fluid: a promising bio-marker of blastocyst developmental potential in women undergoing assisted reproductive technology.

BACKGROUND: Cell-free mitochondrial DNA (cf-mtDNA) in body fluids has attracted much attention for the purpose of monitoring disease because of the clinical advantages. This study investigated whether the cf-mtDNA content in human follicular fluid samples was associated with oocyte and embryo developmental competence. METHODS: We collected 225 individual follicular fluid samples from 92 patients undergoing conventional in vitro fertilization (n = 53) or intracytoplasmic sperm injection (n = 39). cf-mtDNA and cell-free nuclear DNA (cf-nDNA) were measured using real-time quantitative PCR for the ND1 and ß-globin genes. Multivariate logistic regression and linear regression were used to analyze data. RESULTS: The relative cf-mtDNA content (cf-ND1/cf-ß-globin ratio) in follicular fluid was significantly lower in the group showing blastocyst development than in the non-blastocyst group (P = 0.030). Additionally, the relative cf-mtDNA content was significantly and positively correlated with the age of the female patient (P = 0.009), while the relative cf-mtDNA content for older women (≥38 years old) with anti-Müllerian hormone (AMH) ≤1.1 ng/ml was significantly higher than in those with AMH > 1.1 ng/ml (P <0.05). The cf-nDNA content was significantly positively correlated with the antral follicle count (P = 0.012), and significantly negatively correlated with both the number of days of stimulation and the total dose of gonadotropin administration (P = 0.039 and P = 0.015, respectively). Neither cf-mtDNA nor cf-nDNA levels in follicular fluid were associated with oocyte maturation, fertilization, or Day 3 embryo morphological scoring. CONCLUSIONS: The relative cf-mtDNA content in human follicular fluid was negatively correlated with blastulation and positively correlated with the patient age, indicating that it is a promising bio-marker to evaluate oocyte developmental competence.