ATP5D Is a Potential Biomarker for Male Fertility.

Background: Infertility is a global medical and social problem that affects human health and social development. At present, about 15% of couples of the right age in the world are infertile. As all we know, genetic defects are the most likely underlying cause of the pathology. ATP5D is also known as the delta subunit of mitochondrial ATP synthase. Mitochondria maintain sperm vitality, capacitation, acrosome reaction, and DNA integrity through ATP. Mitochondrial damage can trigger energy synthesis disorders, resulting in decreased sperm quality and function or even disappearance. The specific role of ATP5D in regulation of the male reproductive system remains elusive. Methods: In this study, semen from normal and infertile males were collected and their indicators were examined by analysis of routine sperm parameters; ATP5D protein content in semen was examined by ELISA. Singer sequencing was used to detect whether there was a mutated of ATP5D in semen. Meanwhile, ATP5D knockout (KO) and knockin (KI) male mice were selected at 8-12 weeks of age and mated with adult wild-type (WT) female mice for more than two months to assess their fertility and reproductive ability. Morphological changes in tissues such as testes and epididymis were observed by HE staining; spermatozoa were taken from the epididymis of the mice; sperm counts were performed and morphological changes were observed by Diff-Quik staining. Results: The results showed that the expression of ATP5D in infertile males was significantly lower than that in normal males (P < 0.001) and the normal morphology rate of spermatozoa was much lower than that of normal males, and the sequencing results showed no mutations. The animal reproductive experiments showed no significant changes in the number of fertility in KO/KI mice compared with WT mice, but the duration of fertility was significantly longer (P = 0.02). The testicular cells in KO mice were loosely arranged and disorganized, the lumen was larger, the interstitial cells were atrophied, and the number of spermatozoa was reduced and the malformation rate was higher in WT males. This suggests that ATP5D is an essential protein for sperm formation and fertility in male mice and may be used as a biomarker of male fertility. Conclusion: This study found ATP5D correlated with male infertility and the expression levels were significantly reduced in the seminal plasma of all male infertile patients without gene mutations. KO male significantly prolonged fertility time and impaired testicular histomorphology. This suggests that ATP5D may be associated with spermatogenic function and fertility in male mice and may be used as a biomarker for male fertility. Future studies are required to elucidate the potential mechanisms. The trial registration number is KLL-2021-266.

Miracle fruit seed as a potential supplement for the treatment of learning and memory disorders in Alzheimer's disease.

Currently, the treatment of Alzheimer's disease (AD) is still at the stage of symptomatic treatment due to lack of effective drugs. The research on miracle fruit seeds (MFSs) has focused on lipid-lowering and antidiabetic effects, but no therapeutic effects have been reported in AD. The purpose of this study was to provide data resources and a potential drug for treatment of AD. An AD mouse model was established and treated with MFSs for 1 month. The Morris water maze test was used to assess learning memory function in mice. Nissl staining was used to demonstrate histopathological changes. MFSs were found to have therapeutic implications in the AD mouse model, as evidenced by improved learning memory function and an increase in surviving neurons. To explore the mechanism of MFSs in treating AD, network pharmacological approaches, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and molecular docking studies were carried out. Based on the network pharmacology strategy, 74 components from MFS corresponded to 293 targets related to the AD pathology. Among these targets, AKT1, MAPK3, ESR1, PPARG, PTGS2, EGFR, PPARA, CNR1, ABCB1, and MAPT were identified as the core targets. According to the relevant number of core targets, cis-8-octadecenoic acid, cis-10-octadecenoic acid, 2-dodecenal, and tetradecane are likely to be highly correlated with MFS for AD. Enrichment analysis indicated the common targets mainly enriched in AD and the neurodegeneration-multiple disease signaling pathway. The molecular docking predictions showed that MFSs were stably bound to core targets, specifically AKT1, EGFR, ESR1, PPARA, and PPARG. MFSs may play a therapeutic role in AD by affecting the insulin signaling pathway and the Wnt pathway. The findings of this study provide potential possibilities and drug candidates for the treatment of AD.

Lu Tong Ke Li protects neurons from injury by regulating inflammation in rats with brain trauma.

Currently, there is no effective therapy for traumatic brain injury (TBI). Therefore, this study was conducted to determine the protective effect of Lu Tong Ke Li (LTKL), a Chinese medicine, for TBI in experimental animals. The TBI rat model was induced using the modified Feeney's protocol. The rats were divided into four groups: Sham group, Control group, LTKL  lower-dose group (LTL, 2 g/kg/day, p.o.), and LTKL higher-dose group (LTH, 4 g/kg/day, p.o.). The Neurological Severity Score (NSS) was used to examine neurological function. Magnetic resonance imaging was performed to check the brain tissue lesions in rats. Cell apoptosis in the damaged area was evaluated using the Terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling assay. Reverse-transcription polymerase chain reaction was used to investigate the expression of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin 1ß (IL-1ß), and interleukin 10 (IL-10). The TBI rat model was successfully constructed. Neurological function was enhanced at 14, 21, and 28 days post TBI in the LTH groups, indicated by gradually decreased NSS scores. Administration of LTH led to fewer brain defects in the damaged area, and the number of apoptosis cells in the brain injury area markedly decreased. LTKL treatment led to upregulation of IL-10 expression and downregulation of TNF-α and IL-1ß expressions at the molecular level. LTKL can improve the neurobehavior of TBI. The neuroprotective effect was probably related to regulation of inflammation cytokines. Our results provide crucial evidence of the potentially useful application of LTKL in the therapy of TBI in clinic practice in the future.

An integrated analysis of hypoxic-ischemic encephalopathy-related cell sequencing outcomes via genes network construction.

Hypoxic-ischemic encephalopathy (HIE) is one of the main causes of morbidity and severe neurological deficits in neonates. This study aimed to find core genes and their potential roles in HIE with the help of single-cell sequencing (SCS) technology and genes network construction. We collected and screened an HIE genes data set from the Pubmed database to analyze differential expression, and the differential values of genes were ≥3 or ≤-3 in gene expression. We constructed a protein-protein interaction (PPI) network by the string, which was also verified by Cytoscape 3.8.2. Functional enrichment analysis was performed to determine the characteristics and pathways of the core genes. We examined two meaningful papers and integrated all genes by SCS, which were classified into 12,093 genes without duplicates, 217 shared genes, and 11,876 distinct genes. Among 217 genes, the signal transducer and activator of transcription (STAT) family was the most targeted gene in the PPI network. Moreover, Gene Ontology and Kyoto encyclopedia of genes and genome analysis showed that the process in response to virus and the JAK-STAT signaling pathway play significant roles in HIE. We also found that 54 screened genes were highly expressed, while three genes (B2M, VIM, and MRPS30) were different in the heat map and differential genes expression exhibition. VIM, as an essential portion of the brain's cytoskeleton, is closely linked to STAT and neurologic development. From the findings of SCS and bioinformatics predictive analytics model, our outcomes provided a better understanding of the roles of STAT, the JAK-STAT signaling pathway, and VIM, which can pave an alternative avenue for further studies on HIE progression.

Single cell sequencing technology and its application in Hypoxic ischemic encephalopathy research.

Hypoxic ischemic encephalopathy (HIE) is the common etiology of neonatal morbidity and mortality, which exerts a negative seriously influence for the growth and development of children, and even threatens their life. Therapeutic methods are timely not adopted, it will cause serious irreversible damage to the neonatal nervous system. As no promising therapeutic strategies exist currently, it is important to elucidate the pathological mechanism for HIE, which requires us to explore the nucleic acid molecules, protein, and cell function in HIE patients, and to understand the process of the onset and progression, then research and invent better treatment methods and therapeutic drugs. Single cell sequencing (SCS) exhibits an distinctive advantages in cells research because of the particularity of each cell. This method solves an puzzle about heterogeneit, which could not be solved with multi cell sample research, and provides a new idea and perspective for the un-elucidated and events further analyzed, such as the behaviors, mechanisms and the relationship between single cell and organism in cell population. It also plays an extremely significant role in the basic research and precision medicine. Some studies have suggested that SCS serves a vital function in the study of HIE. Therefore, this review is aim to elaborate SCS and hypoxic-ischemic brain injury, and trace the role of microglia in HIE, and prospect its unknown and undiscovered mechanism by SCS.