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

OBJECTIVE: To explore the functional roles and underlying mechanisms of neferine in the context of angiotensin II (Ang II)-induced hypertension and vascular dysfunction. METHODS: Male mice were infused with Ang II to induce hypertension and randomly divided into treatment groups receiving neferine or a control vehicle based on baseline blood pressure using a random number table method. The hypertensive mouse model was constructed by infusing Ang II via a micro-osmotic pump (500 ng/kg per minute), and neferine (0.1, 1, or 10 mg/kg), valsartan (10 mg/kg), or double distilled water was administered intragastrically once daily for 6 weeks. A non-invasive blood pressure system, ultrasound, and hematoxylin and eosin staining were performed to assess blood pressure and vascular changes. RNA sequencing and network pharmacology were employed to identify differentially expressed transcripts (DETs) and pathways. Vascular ring tension assay was used to test vascular function. A7R5 cells were incubated with neferine for 24 h and then treated with Ang II to record the real-time Ca²⁺ concentration by confocal microscope. Immunohistochemistry (IHC) and Western blot were used to evaluate vasorelaxation, calcium, and the extracellular signal-regulated kinase (ERK)1/2 pathway. RESULTS: Neferine treatment effectively mitigated the elevation in blood pressure, pulse wave velocity, aortic thickening in the abdominal aorta of Ang II-infused mice (P<0.05). RNA sequencing and network pharmacology analysis identified 355 DETs that were significantly reversed by neferine treatment, along with 25 potential target genes, which were further enriched in multiple pathways and biological processes, such as ERK1 and ERK2 cascade regulation, calcium pathway, and vascular smooth muscle contraction. Further investigation revealed that neferine treatment enhanced vasorelaxation and reduced Ca²⁺-dependent contraction of abdominal aortic rings, independent of endothelium function (P<0.05). The underlying mechanisms were mediated, at least in part, via suppression of receptor-operated channels, store-operated channels, or voltage-operated calcium channels. Neferine pre-treatment demonstrated a reduction in intracellular Ca²⁺ release in Ang II stimulated A7R5 cells. IHC staining and Western blot confirmed that neferine treatment effectively attenuated the upregulation of p-ERK1/2 both in vivo and in vitro, which was similar with treatment of ERK1/2 inhibitor PD98059 (P<0.05). CONCLUSIONS Neferine remarkably alleviates Ang II-induced elevation of blood pressure, vascular dysfunction, and pathological changes in the abdominal aorta. This beneficial effect is mediated by the modulation of multiple pathways, including calcium and ERK1/2 pathways.
Animals ; Angiotensin II ; Male ; Benzylisoquinolines/therapeutic use* ; Network Pharmacology ; Blood Pressure/drug effects* ; Sequence Analysis, RNA ; Mice ; Hypertension/chemically induced* ; Mice, Inbred C57BL ; Calcium/metabolism*

OBJECTIVE: The objective of our study was to evaluate the vaccine effectiveness (VE) of the pentavalent rotavirus vaccine (RV5) among < 5-year-old children in three provinces of China during 2020-2024 via a propensity score-matched test-negative case-control study. METHODS: Electronic health records and immunization information systems were used to obtain data on acute gastroenteritis (AGE) cases tested for rotavirus (RV) infection. RV-positive cases were propensity score matched with RV-negative controls for age, visit month, and province. RESULTS: The study included 27,472 children with AGE aged 8 weeks to 4 years at the time of AGE diagnosis; 7.98% (2,192) were RV-positive. The VE (95% confidence interval, CI) of 1-2 and 3 doses of RV5 against any medically attended RV infection (inpatient or outpatient) was 57.6% (39.8%, 70.2%) and 67.2% (60.3%, 72.9%), respectively. Among children who received the 3rd dose before turning 5 months of age, 3-dose VE decreased from 70.4% (53.9%, 81.1%) (< 5 months since the 3rd dose) to 63.0% (49.1%, 73.0%) (≥ 1 year since the 3rd dose). The three-dose VE rate was 69.4% (41.3%, 84.0%) for RVGE hospitalization and 57.5% (38.9%, 70.5%) for outpatient-only medically attended RVGE. CONCLUSION Three-dose RV5 VE against rotavirus gastroenteritis (RVGE) in children aged < 5 years was higher than 1-2-dose VE. Three-dose VE decreased with time since the 3rd dose in children who received the 3rd dose before turning five months of age, but remained above 60% for at least one year. VE was higher for RVGE hospitalizations than for medically attended outpatient visits.
Humans ; Rotavirus Vaccines/immunology* ; China/epidemiology* ; Case-Control Studies ; Child, Preschool ; Infant ; Rotavirus Infections/epidemiology* ; Male ; Propensity Score ; Female ; Vaccine Efficacy ; Gastroenteritis/virology* ; Vaccines, Attenuated ; Rotavirus

Objective To comparatively analyze the clinical characteristics of primary bilateral macronodular adrenal hyperplasia(PBMAH)and adrenal cortisol-producing Adenoma(CPA),and enhance the understanding of two diseases.Methods The clinical data of 85 PBMAH patients(PBMAH group)and 195 CPA patients(CPA group)diagnosed at Department of Endocrinology,the First Medical Center of Chinese PLA General Hospital,from September 2014 to August 2024 were retrospectively analyzed.The demographic characteristics,comorbidities,biochemical indicators,adrenocorticotropic hormone-cortisol(ACTH-F)levels,and adrenal imaging features and treatment conditions were compared between the two groups.Results(1)General characteristics:Compared with CPA group,PBMAH group had older age at diagnosis and a higher proportion of male patients.(2)Clinical characteristics:Compared with CPA group,PBMAH group had a longer disease duration,a higher proportion of subclinical Cushing's syndrome(CS),and a higher proportion of hypertension,impaired glucose tolerance/diabetes,bone mass reduction or osteoporosis,with higher serum potassium levels,and the differences were statistically significant(P<0.01).(3)Hormone levels:Both PBMAH and CPA groups showed ACTH-F rhythm disorder,significantly increased cortisol levels and suppressed ACTH.Compared with PBMAH group,CPA group had stronger autonomous cortisol secretion ability,manifested by increased midnight serum cortisol(F0:00),16:00 serum cortisol(F16:00),24-hour urinary free cortisol(24 h UFC)levels and lower 8:00 serum ACTH(ACTH8:00)and 16:00 serum ACTH(ACTH16:00)(P<0.01).After low-dose dexamethasone suppression test(LDDST),CPA group showed lower suppression rates of ACTH and cortisol,and higher proportions of paradoxical elevation in serum cortisol and 24 h UFC compared with PBMAH(P<0.01).Conclusions PBMAH has a longer disease course and higher proportions of comorbid metabolic disorders than CPA,mostly manifested as subclinical Cushing's syndrome.CPA has stronger autonomous cortisol secretion ability,with cortisol less likely to be suppressed after LDDST and more obvious paradoxical elevation of cortisol and 24 h UFC.

Objective To investigate the clinical and pathological characteristics of adrenal cortical carcinoma(ACC),compare differences between hypercortisolism and non-functional ACC,and assess the diagnostic value of indicators such as Ki-67 index.Methods The clinical data of 57 ACC patients admitted to the First Medical Center of Chinese PLA General Hospital from January 2015 to March 2025 were retrospectively analyzed.According to the results of endocrine function assessment,47 of these patients were divided into hypercortisolism group(n=19)and non-functional group(n=28).The differences in clinical and pathological characteristics between the two groups were compared,and non-parametric tests and Spearman correlation analysis were used to explore the relationship between Ki-67 index and tumor stage as well as imaging features.Results Among the 57 patients,there were 20 males and 37 females,with a male-to-female ratio of 1:1.85.The age ranged from 16 to 76 years,and the age at diagnosis was(48.7±13.3)years.The tumor diameter was(10.53±4.14)cm.The tumors were located on the right side in 12 cases(21.1%),on the left side in 34 cases(59.6%),and bilaterally in 11 cases(19.3%).Among them,16 cases(28.1%)were complicated with glucose metabolism disorders,31 cases(54.3%)had hypertension,and 20 cases(35.1%)had hypokalemia.According to ENSAT staging,there were 0 cases in stage Ⅰ,15 cases(26.3%)in stage Ⅱ,24 cases(42.1%)in stage Ⅲ,and 18 cases(31.6%)in stage Ⅳ.Endocrine function assessment was completed in 47 of the 57 patients,including 28 cases(59.6%)of non-functional ACC and 19 cases(40.4%)of hypercortisolism(including 1 case of hypercortisolism combined with increased sex hormone secretion).Compared with non-functional group,hypercortisolism group had a significantly higher prevalence of hypertension(P=0.014),later ENSAT stage(P=0.010),and a higher proportion of hypervascularization(P=0.048).The median Ki-67 index was 20%(10%-40%),showing no significant correlation with either the maximum tumor diameter or SUVmax value,but it was related to ENSAT staging,with Ki-67 index in stageⅣ patients being significantly higher than that in stage Ⅱ(P=0.032).Immunohistochemistry results showed that the positive rate of Inhibin-α was 84.8%,and the positive rate of Melan-A was 40.9%.Conclusions ACC is a rare malignant endocrine tumor.ACC patients with hypercortisolism are more likely to be complicated with hypertension,have later staging,and more common hypervascular manifestations.Clinically,their endocrine function should be prioritized for assessment,and more active treatment strategies should be adopted.Diagnosis should be combined with imaging characteristics(such as hypervascularization)and immunohistochemical indicators(Ki-67,Inhibin-α,Melan-A).The significant increase in Ki-67 is in the advanced stage can serve as an important prognostic indicator to guide individualized treatment.

The engineering application of microbially induced carbonate precipitation(MICP)is limited by pH-dependent conformational dynamics of urease.Focusing on the α-subunit urease from Sporosarcina pasteurii,this study integrated conductivity experiments and constant-pH molecular dynamics simulations to analyze active site conformational dynamics and catalytic function across pH 3-11.Results showed that under neutral conditions(pH 7-8),key histidine residues(HIS139/HIS249)exhibited minimal dis-placement(＜0.5 ?),the longest hydrogen bond lifetime(＞8 ps),highest conformational stability(root mean square deviation,RMSD:0.15-0.18 nm),and optimal catalytic activity(conductivity change rate:0.03 mS/cm·min-1,CaCO3 precipitation:3.84 g).Extreme pH(pH 3/11)induced structural collapse(displacement up to 1.8 ?)and complete activity loss.Simulations revealed that neutral pH sta-bilizes a protonation-dependent cooperative allosteric network by maintaining active site cavity volume(～120 ?3)and moderate conformational coherence(correlation coefficient～0.8).This work deciphers the molecular mechanism of pH-regulated urease dynamics through protonation states,providing theoreti-cal support for MICP applications in acidic mine tailing remediation and alkaline soil stabilization.

Objective:To construct and validate a three-level early warning model of moderate to severe ovarian hyperstimulation syndrome (OHSS) in assisted reproductive technology.Methods:A case-control study was conducted. Totally 10 181 infertility patients who underwent in vitro fertilization treatment in Reproductive Medicine Center, Women and Children's Medical Center Affiliated to Guangzhou Medical University from April 2013 to April 2023 were retrospectively analyzed and divided into modeling group (8 145 cases) and validation group (2 036 cases) by random number table method. The clinical data of the two groups were analyzed, and the risk factors affecting the occurrence of moderate and severe OHSS after oocytes retrieval were screened by multi-factor logistic regression analysis. The early warning model was established and the column diagram was drawn at three nodes which were before ovarian stimulation, before trigger and 3 d after oocyte retrieval. The receiver operating characteristic (ROC) curve and calibration curve were used to verify the models. Results:The antral follicle count (AFC, OR=1.045, 95% CI: 1.020-1.071, P<0.001), anti-Müllerian hormone (AMH)>3.36 μg/L ( OR=7.135, 95% CI: 2.084-24.432, P=0.002) and number of cycles ( OR=0.149, 95% CI: 0.022-1.026, P=0.049) were included in the pre-stimulation prediction model. AFC ( OR=1.046, 95% CI: 1.018-1.074, P=0.001), AMH>3.36 μg/L ( OR=5.780, 95% CI: 1.661-20.116, P=0.006), gonadotropin releasing hormone-agonist protocols ( OR=3.895, 95% CI=1.913-7.931, P<0.001), estrogen peak≥18 350 pmol/L ( OR=2.258, 95% CI: 1.092-4.666, P=0.028), the number of follicles with a diameter of ≥10 mm>20 ( OR=2.377, 95% CI: 1.092-5.172, P=0.029) were included in the pre-trigger prediction model. AMH>3.36 μg/L ( OR=8.374, 95% CI: 2.417-29.019, P=0.001), estrogen peak≥18 350 pmol/L ( OR=3.947, 95% CI: 1.533-10.167, P=0.004), total number of oocytes retrived ( OR=1.042, 95% CI: 0.996-1.090, P=0.025), abdominal distension ( OR=60.181, 95% CI: 22.515-160.854, P<0.001), fresh transplantation ( OR=21.766, 95% CI: 7.119-66.544, P<0.001), human chorionic gonadotropin trigger ( OR=17.752, 95% CI: 3.993-78.924, P<0.001) were included in the prediction model of 3 d after oocyte retrieval. The areas under ROC curves of the three models were 0.830 (95% CI: 0.782-0.878), 0.859 (95% CI: 0.812-0.906) and 0.948 (95% CI: 0.919-0.977), respectively. The areas under ROC curves of the validation groups of the three models were 0.922 (95% CI: 0.880-0.965), 0.936 (95% CI: 0.886-0.986), and 0.971 (95% CI: 0.938-0.999), respectively. The calibration curve indicated that the early-warning evaluation model has good stability. Conclusion:The three-level early warning model of moderate and severe OHSS has good differentiation, reliable predictability and clinical practicability, which is conducive to the dynamic and continuous assessment of the risk of moderate and severe OHSS, adjustment of treatment plan at any time, and timely adoption of effective preventive measures.

Objective:To construct and validate a three-level early warning model of moderate to severe ovarian hyperstimulation syndrome (OHSS) in assisted reproductive technology.Methods:A case-control study was conducted. Totally 10 181 infertility patients who underwent in vitro fertilization treatment in Reproductive Medicine Center, Women and Children's Medical Center Affiliated to Guangzhou Medical University from April 2013 to April 2023 were retrospectively analyzed and divided into modeling group (8 145 cases) and validation group (2 036 cases) by random number table method. The clinical data of the two groups were analyzed, and the risk factors affecting the occurrence of moderate and severe OHSS after oocytes retrieval were screened by multi-factor logistic regression analysis. The early warning model was established and the column diagram was drawn at three nodes which were before ovarian stimulation, before trigger and 3 d after oocyte retrieval. The receiver operating characteristic (ROC) curve and calibration curve were used to verify the models. Results:The antral follicle count (AFC, OR=1.045, 95% CI: 1.020-1.071, P<0.001), anti-Müllerian hormone (AMH)>3.36 μg/L ( OR=7.135, 95% CI: 2.084-24.432, P=0.002) and number of cycles ( OR=0.149, 95% CI: 0.022-1.026, P=0.049) were included in the pre-stimulation prediction model. AFC ( OR=1.046, 95% CI: 1.018-1.074, P=0.001), AMH>3.36 μg/L ( OR=5.780, 95% CI: 1.661-20.116, P=0.006), gonadotropin releasing hormone-agonist protocols ( OR=3.895, 95% CI=1.913-7.931, P<0.001), estrogen peak≥18 350 pmol/L ( OR=2.258, 95% CI: 1.092-4.666, P=0.028), the number of follicles with a diameter of ≥10 mm>20 ( OR=2.377, 95% CI: 1.092-5.172, P=0.029) were included in the pre-trigger prediction model. AMH>3.36 μg/L ( OR=8.374, 95% CI: 2.417-29.019, P=0.001), estrogen peak≥18 350 pmol/L ( OR=3.947, 95% CI: 1.533-10.167, P=0.004), total number of oocytes retrived ( OR=1.042, 95% CI: 0.996-1.090, P=0.025), abdominal distension ( OR=60.181, 95% CI: 22.515-160.854, P<0.001), fresh transplantation ( OR=21.766, 95% CI: 7.119-66.544, P<0.001), human chorionic gonadotropin trigger ( OR=17.752, 95% CI: 3.993-78.924, P<0.001) were included in the prediction model of 3 d after oocyte retrieval. The areas under ROC curves of the three models were 0.830 (95% CI: 0.782-0.878), 0.859 (95% CI: 0.812-0.906) and 0.948 (95% CI: 0.919-0.977), respectively. The areas under ROC curves of the validation groups of the three models were 0.922 (95% CI: 0.880-0.965), 0.936 (95% CI: 0.886-0.986), and 0.971 (95% CI: 0.938-0.999), respectively. The calibration curve indicated that the early-warning evaluation model has good stability. Conclusion:The three-level early warning model of moderate and severe OHSS has good differentiation, reliable predictability and clinical practicability, which is conducive to the dynamic and continuous assessment of the risk of moderate and severe OHSS, adjustment of treatment plan at any time, and timely adoption of effective preventive measures.

The engineering application of microbially induced carbonate precipitation(MICP)is limited by pH-dependent conformational dynamics of urease.Focusing on the α-subunit urease from Sporosarcina pasteurii,this study integrated conductivity experiments and constant-pH molecular dynamics simulations to analyze active site conformational dynamics and catalytic function across pH 3-11.Results showed that under neutral conditions(pH 7-8),key histidine residues(HIS139/HIS249)exhibited minimal dis-placement(＜0.5 ?),the longest hydrogen bond lifetime(＞8 ps),highest conformational stability(root mean square deviation,RMSD:0.15-0.18 nm),and optimal catalytic activity(conductivity change rate:0.03 mS/cm·min-1,CaCO3 precipitation:3.84 g).Extreme pH(pH 3/11)induced structural collapse(displacement up to 1.8 ?)and complete activity loss.Simulations revealed that neutral pH sta-bilizes a protonation-dependent cooperative allosteric network by maintaining active site cavity volume(～120 ?3)and moderate conformational coherence(correlation coefficient～0.8).This work deciphers the molecular mechanism of pH-regulated urease dynamics through protonation states,providing theoreti-cal support for MICP applications in acidic mine tailing remediation and alkaline soil stabilization.