Recent studies have shown that shorter periods of ejaculatory abstinence may enhance certain sperm parameters, but the molecular mechanisms underlying these improvements are still unclear. This study explored whether reduced abstinence periods could improve semen quality, particularly for use in assisted reproductive technologies (ART). We analyzed semen samples from men with normal sperm counts ( n = 101) and those with low sperm motility or concentration ( n = 53) after 3-7 days of abstinence and then after 1-3 h of abstinence, obtained from the Reproductive & Genetic Hospital of CITIC-Xiangya (Changsha, China). Physiological and biochemical sperm parameters were evaluated, and the dynamics of transfer RNA (tRNA)-derived fragments (tRFs) were analyzed using deep RNA sequencing in five consecutive samples from men with normal sperm counts. Our results revealed significant improvement in sperm motility and a decrease in the DNA fragmentation index after the 1- to 3-h abstinence period. Additionally, we identified 245 differentially expressed tRFs, and the mitogen-activated protein kinase (MAPK) signaling pathway was the most enriched. Further investigations showed significant changes in tRF-Lys-TTT and its target gene mitogen-activated protein kinase kinase 2 ( MAP2K2 ), which indicates a role of tRFs in improving sperm function. These findings provide new insights into how shorter abstinence periods influence sperm quality and suggest that tRFs may serve as biomarkers for male fertility. This research highlights the potential for optimizing ART protocols and improving reproductive outcomes through molecular approaches that target sperm function.
Male ; Humans ; Spermatozoa/metabolism* ; RNA, Transfer/genetics* ; Sperm Motility/genetics* ; Adult ; Semen Analysis ; Sexual Abstinence/physiology* ; Sperm Count ; DNA Fragmentation

The aryl hydrocarbon receptor (AhR) plays a crucial role in regulating many physiological processes. Activating the AhR-CYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases. Here, a practical in situ cell-based fluorometric assay was constructed to screen AhR-CYP1A1 axis modulators, via functional sensing of CYP1A1 activities in live cells. Firstly, a cell-permeable, isoform-specific enzyme-activable fluorogenic substrate for CYP1A1 was rationally constructed for in-situ visualizing the dynamic changes of CYP1A1 function in living systems, which was subsequently used for discovering the efficacious modulators of the AhR-CYP1A1 axis. Following screening of a compound library, LAC-7 was identified as an efficacious activator of the AhR-CYP1A1 axis, which dose-dependently up-regulated the expression levels of both CYP1A1 and AhR in multiple cell lines. LAC-7 also suppressed macrophage M1 polarization and reduced the levels of inflammatory factors in LPS-induced bone marrow-derived macrophages. Animal tests showed that LAC-7 could significantly mitigate DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice, and markedly reduced the levels of multiple inflammatory factors. Collectively, an optimized fluorometric cell-based assay was devised for in situ functional imaging of CYP1A1 activities in living systems, which strongly facilitated the discovery of efficacious modulators of the AhR-CYP1A1 axis as novel anti-inflammatory agents.

Despite therapy with potent antiviral agents, chronic hepatitis B (CHB) patients remain at high risk of hepatocellular carcinoma (HCC). While metabolites have been rediscovered as active drivers of biological processes including carcinogenesis, the specific metabolites modulating HCC risk in CHB patients are largely unknown. Here, we demonstrate that baseline plasma from CHB patients who later developed HCC during follow-up exhibits growth-promoting properties in a case-control design nested within a large-scale, prospective cohort. Metabolomics analysis reveals a reduction in long-chain acylcarnitines (LCACs) in the baseline plasma of patients with HCC development. LCACs preferentially inhibit the proliferation of HCC cells in vitro at a physiological concentration and prevent the occurrence of HCC in vivo without hepatorenal toxicity. Uptake and metabolism of circulating LCACs increase the intracellular level of acetyl coenzyme A, which upregulates histone H3 Lys14 acetylation at the promoter region of KLF6 gene and thereby activates KLF6/p21 pathway. Indeed, blocking LCAC metabolism attenuates the difference in KLF6/p21 expression induced by baseline plasma of HCC/non-HCC patients. The deficiency of circulating LCACs represents a driver of HCC in CHB patients with viral control. These insights provide a promising direction for developing therapeutic strategies to reduce HCC risk further in the antiviral era.

Notum, a negative feedback regulator of the Wnt signaling, has emerged as a promising target for treating glucocorticoid-induced osteoporosis (GIOP). This study showcases an efficient strategy for discovering the anti-Notum constituents from herbal medicines (HMs) as novel anti-GIOP agents. Firstly, a rapid-responding near-infrared fluorogenic substrate for Notum was rationally engineered for high-throughput identifying the anti-Notum HMs. The results showed that Bu-Gu-Zhi (BGZ), a known anti-osteoporosis herb, potently inhibited Notum in a competitive-inhibition manner. To uncover the key anti-Notum constituents in BGZ, an efficient strategy was adapted via integrating biochemical, phytochemical, computational, and pharmacological assays. Among all identified BGZ constituents, three furanocoumarins were validated as strong Notum inhibitors, while 5-methoxypsoralen (5-MP) showed the most potent anti-Notum activity and favorable safety profiles. Mechanistically, 5-MP acted as a competitive inhibitor of Notum via creating strong hydrophobic interactions with Trp128 and Phe268 in the catalytic cavity of Notum. Cellular assays showed that 5-MP remarkably promoted osteoblast differentiation and activated Wnt signaling in dexamethasone (DXMS)-challenged MC3T3-E1 osteoblasts. In dexamethasone-induced osteoporotic mice, 5-MP strongly elevated bone mineral density (BMD) and improved cancellous and cortical bone thickness. Collectively, this study constructs a high-efficient platform for discovering key anti-Notum constituents from HMs, while 5-MP emerges as a promising anti-GIOP agent.

Human carboxylesterase 2A (hCES2A) plays pivotal roles in prodrug activation and hydrolytic metabolism of ester-bearing chemicals. Targeted inhibition of intestinal hCES2A represents a feasible strategy to mitigate irinotecan-triggered gut toxicity (ITGT), but the orally active, selective, and efficacious hCES2A inhibitors are rarely reported. Here, a novel drug-like hCES2A inhibitor was developed via three rounds of structure-based drug design (SBDD) and structural optimization. Initially, donepezil was identified as a moderate hCES2A inhibitor from 2000 US Food and Drug Administration (FDA)-approved drugs. Following two rounds of SBDD and structural optimization, a donepezil derivative (B7) was identified as a strong reversible hCES2A inhibitor. Subsequently, nine B7 carbamates were rationally designed, synthesized and biologically assayed. Among all synthesized carbamates, C3 showed the most potent time-dependent inhibition on hCES2A (IC₅₀ = 0.56 nmol/L), excellent specificity and favorable drug-like properties. C3 could covalently modify the catalytic serine of hCES2A with high selectivity, while this agent also showed favorable safety profiles, high intestinal exposure, and impressive effects for ameliorating ITGT in both human intestinal organoids and tumor-bearing mice. Collectively, this study showcases a rational strategy for developing drug-like and serine-targeting covalent inhibitors against target serine hydrolase(s), while C3 emerges as a promising orally active drug candidate for ameliorating ITGT.

Hypoxic injury (HI) in the prenatal period often causes neonatal neurological disabilities. Due to the difficulty in obtaining clinical samples, the molecular and cellular mechanisms remain unclear. Here we use vascularized cerebral organoids to investigate the hypoxic injury phenotype and explore the intercellular interactions between vascular and neural tissues under hypoxic conditions. Our results indicate that fused vascularized cerebral organoids exhibit broader hypoxic responses and larger decreases in panels of neural development-related genes when exposed to low oxygen levels compared to single cerebral organoids. Interestingly, vessels also exhibit neural protective effects on T-box brain protein 2⁺ intermediate progenitors (IPs), which are markedly lost in HI cerebral organoids. Furthermore, we identify the role of bone morphogenic protein signaling in protecting IPs. Thus, this study has established an in vitro organoid system that can be used to study the contribution of vessels to brain injury under hypoxic conditions and provides a strategy for the identification of intervention targets.
Organoids/pathology* ; Animals ; Mice ; Hypoxia, Brain/metabolism* ; Brain/blood supply* ; Neurons/metabolism*

Objective To retrieve,evaluate and summarize the best evidence of early mobilization in awake patients on extracorporeal membrane oxygenation,and to provide a reference for clinical practice.Methods UpToDate,BMJ Best Practice,Registered Nurses'Association of Ontario,National Guideline Clearinghouse,National Institute for Health and Care Excellence,Yimaitong,Joanna Briggs Institute Library,Cochrane Library,CINAHL,PubMed,SinoMed,CNKI,Wanfang Database,Vip Database and Extracorporeal Life Support Organization Website were researched to collect the literature,including clinical guidelines,expert consensuses,evidence summaries,systematic reviews,and well-designed original studies.The time limit for retrieval was until June 2023.The quality of literature and the level of evidence were evaluated by the evaluation criteria and evidence grading system of J BI Evidence-Based Health Care Center.Results 14 pieces of the literature were included,including 2 clinical guidelines,4 expert consensuses,5 systematic reviews,2 cohort studies and 1 case series.Totally 33 pieces of evidence were summarized,covering 7 aspects:adaptation conditions for the implementation of awake ECMO,team composition,comprehensive assessment,pre-mobilization preparation,mobilization content,prevention and control of adverse events,and effect evaluation.Conclusion The study summarizes the best evidence of early mobilization in awake patients on extracorporeal membrane oxygenation.It is suggested that medical institutions establish a professional team for the early mobilization of awake ECMO patients,apply the best evidence to standardize the early mobilization process,and formulate an individualized mobilization program.

Objective:To explore the value of salivary gland imaging based on deep learning and Delta radiomics in assessing salivary gland injury after 131I treatment in post-thyroidectomy thyroid cancer patients. Methods:A retrospective analysis on 223 patients (46 males, 177 females, age(47.7±14.0) years ) with papillary thyroid cancer, who underwent total thyroidectomy and 131I treatment in Affiliated Hospital of Guilin Medical University between December 2019 and January 2022, was conducted. All patients underwent salivary gland 99Tc mO 4- imaging before and after 131I therapy. The patients were categorized according to salivary gland function based on 99Tc mO 4- imaging results (normal salivary gland vs salivary gland injury), and divided into training and test sets in a ratio of 7∶3. A ResNet-34 neural network model was trained using images at the time of maximum salivary gland radioactivity and those based on background radioactivity counts for structured image feature data. The Delta radiomics approach was then used to subtract the image feature values of the two periods, followed by feature selection through t-test, correlation analysis, and the least absolute shrinkage and selection operator( LASSO) algorithm, to develop logistic regression (LR), support vector machine (SVM), and K-nearest neighbor (KNN) predictive models. The diagnostic performance of 3 models for salivary gland function on the test set was compared with that of the manual interpretation. The AUCs of the 3 models on the test set were compared (Delong test). Results:Among the 67 cases of the test set, the diagnostic accuracy of 3 physicians were 89.6%(60/67), 83.6%(56/67), and 82.1%(55/67) respectively, with the time required for diagnosis of 56, 74 and 55 min, respectively. The accuracies of LR, SVM, and KNN models were 91.0%(61/67), 86.6%(58/67), and 82.1%(55/67), with the required times of 12.5, 15.3 and 17.9 s, respectively. All 3 radiomics models demonstrated good classification and predictive capabilities, with AUC values for the training set of 0.972, 0.965, and 0.943, and for the test set of 0.954, 0.913, and 0.791, respectively. There were no significant differences among the AUC values for the test set ( z values: 0.72, 1.18, 1.82, all P>0.05). Conclusion:The models based on deep learning and Delta radiomics possess high predictive value in assessing salivary gland injury following 131I treatment after surgery in patients with thyroid cancer.

Objective To analyze the relationship of serum S100 calcium binding protein A4(S100A4)and pentraxin-3(PTX3)levels with left atrial appendage thrombosis in patients with NVAF.Methods A total of 120 elderly NVAF patients treated in our hospital from March 2020 to March 2023 were enrolled in this study.According to their echocardiograms,they were divided into a left atrial appendage thrombosis group(40 cases)and a non-thrombosis group(80 cases).Serum S100A4 and PTX3 levels were detected.Spearman correlation analysis was applied to ana-lyze the relationship between serum S100A4 and PTX3 levels and left atrial appendage thrombo-sis.Logistic regression analysis was conducted to analyze the factors affecting left atrial appendage thrombosis.Results The serum levels of S100A4 and PTX3 were higher in the thrombosis group than the non-thrombosis group(P＜0.01).The serum levels of S100A4 and PTX3 were positively correlated with left atrial appendage thrombosis(r=0.497,P=0.000;r=0.555,P=0.000).Heart failure,CHA2DS2-VASc score,B-type natriuretic peptide,uric acid,S100A4 and PTX3 were risk factors for left atrial appendage thrombosis in NVAF patients(P＜0.05,P＜0.01).Combination of serum S100A4 and PTX3 in predicting left atrial appendage thrombosis formation in NVAF patients had an AUC value of of 0.949(95％CI:0.893-0.981).Conclusion Serum S100A4 and PTX3 levels are increased in NVAF patients,they are related to left atrial appendage thrombosis,and their serum levels have certain predictive value for left atrial appendage thrombosis.