Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

Aim To investigate the effect of oroxylin A(OA)on apoptosis in Ishikawa cell line of endometrial cancer and the underlying mechanism through the phosphatidylinositol-3 kinase/protein kinase B(PI3K/AKT)signaling pathway.Methods Ishikawa cells were treated with different concentrations of OA(0,4,8,10,12,and 20 μmol·L-1)for 24 h-72 h,the cell viability was detected by CCK-8 assay,apoptosis was detected by flow cytometry,and the protein ex-pression levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),PI3K/AKT,recombinant cytochrome P450 1B1(CYP1B1),and catechol-O-methyltransferase(COMT)were detected by Western blot technique.Results OA inhibited the prolifera-tion of Ishikawa cells in a concentration-and time-de-pendent manner.Compared with the blank control group,the expression of Bax protein increased signifi-cantly,while the expression of Bcl-2 protein decreased significantly with the increase of OA concentration.The expression of COMT protein increased significant-ly,while the expression of CYP1B1 protein decreased significantly.PI3K/AKT:IGF-1(PI3 K agonist)sup-plementation reversed the effect,the expression of COMT protein significantly decreased,and the expres-sion of CYP1B1 protein significantly increased.Con-clusions OA exerts anti-tumor effects in Ishikawa cells of endometrial cancer,which may be related to cell apoptosis mediated by the inhibition of the PI3K/AKT signaling pathway.

Biochemistry,as a fundamental course for science and engineering majors related to biology and chemistry,holds a significant position in the curriculum.The course team at Dalian Minzu University is committed to teaching innovation,adopting the outcome-based education(OBE)concept for teaching de-sign and incorporating ideological and political elements,in order to achieve the dual goals of knowledge transmission and value guidance.The team has established a three-dimensional teaching goal of"knowl-edge,morality,and ability",covering"consolidating core knowledge,cultivating moral sentiment,and enhancing innovation ability".Through a multi-dimensional integrated teaching method of"three integra-tions and five combinations",multiple rounds of teaching practice have been carried out in the applied chemistry major using"classification and specificity of enzyme"as an example.The output of teaching re-sults and survey questionnaires show that students highly recognize the teaching design and its"process-based learning"evaluation method,fully reflecting the student-centered teaching idea.Research has shown that OBE design combined with ideological and political elements can effectively promote students' knowl-edge acquisition,moral growth,and innovation ability improvement in the course of Biochemistry.This teaching design not only helps students construct correct worldviews,outlooks on life,and values,but also significantly enhances their innovative thinking and practical abilities.This teaching design can not only ef-fectively improve the teaching quality of the course,but also provide new perspectives and ideas for the teaching design of Biochemistry,realizing the organic integration of professional knowledge imparting and i-deological and political education,and has certain innovation and practical significance.

A method for simultaneous determination of 21 kinds of Aconitum alkaloids(ATS)in biological specimens and infused liquor using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed.The biological samples were pretreated with methanol-acetonitrile(1∶2,V/V)for protein precipitation,while infused liquors were diluted 100-fold with acetonitrile,followed by centrifugation,and filtration by a 0.22-μm membrane.Chromatographic separation was carried out on an EC-C18 column using gradient elution with the mixture of 10 mmol/L ammonium acetate and 0.2%formic acid as mobile phase A and acetonitrile as mobile phase B.With this method,all the analytes were separated within 9.5 min.The samples were detected in positive ESI mode with dynamic multiple reaction monitoring(MRM)and quantified via external standard calibration.The results showed that the concentrations of the analytes in the range of 2-1000 ng/mL had excellent linearity(R2>0.9992)with the peak area.The developed method was successfully used for detection of 21 kinds of aconitum alkaloids,with limits of detection of 0.5-2 ng/mL,quantification limits of 2-6 ng/mL,intra/inter-day precision≤6.0%,spiked recoveries of 89.4%-100.9%,extraction recoveries of 74.2%-104.4%,and matrix effects ranging from-11.1%to 9.2%in blood/urine.The method was applied to detection of 12 samples from 4 fatal aconite poisoning cases,and all 21 kinds of ATS with total alkaloid concentrations of 0.04-4.18 μg/mL in blood and 154.96-422.83 μg/mL in medicinal liquors were detected.Tissue distribution revealed that the order of concentrations from highest to lowest is as follows:urine(157.22 μg/mL)>gastric contents(51.37 μg/mL)>kidney(21.6 μg/g)>whole blood(4.18 μg/mL)>liver(0.03 μg/g).This method showed many advantages such as simple pretreatment,low detection limits,accurate quantification,broad analyte coverage,and superior anti-interference capability in complex matrices,proving ideal for forensic and toxicological analysis of aconitum alkaloids.

Objective To observe the effect of emodin on lung injury in rats with stroke associated pneumonia(SAP)through regulating Janus kinase 2(JAK2)/signal transducer and activators of transcription 3(STAT3)signaling pathway.Methods The SAP rat model was established by modified thread embolization method combined with intratracheal injection of Pseudomonas aeruginosa method.The successfully modeled rats were randomly divided into model group,emodin low-and high-dose groups and emodin high-dose+coumermycin(JAK2 activator)group,with 12 rats in each group,and another 12 SD rats were selected as the sham-operated group.After the intervention,the partial pressure of arterial blood gas indexes such as carbon dioxide(PaCO2)and partial pressure of oxygen(PaO2)were measured,the ratio of lung wet/dry mass(W/D)was determined,the inflammatory cell count in the bronchoalveolar lavage fluid(BALF)was detected,the histopathological feature in the lung tissues was observed by hematoxylin-eosin(HE)staining and the pathological injury was scored,enzyme-linked immunosorbent assay(ELISA)was used to detect myeloperoxidase(MPO)activity in the lung tissues and interleukin 6(IL-6)and cyclooxygenase 2(COX-2)levels in BALF and serum,and Western Blot was used to detect the expression of JAK2/STAT3 pathway key proteins in lung tissues.Results Compared with the sham-operated group,the model group rats showed that more obvious pathological injury in lung tissues was seen,and PaCO2 value,ratio of lung W/D,white blood cell count and eosinophils count in BALF,pathological injury score,MPO activity in lung tissues,IL-6 and COX-2 levels in BALF and serum,and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissues were significantly increased,and PaO2 value was significantly decreased,the differences being statistically significant(P<0.05);compared with the model group,the emodin low-and high-dose groups rats showed that the pathological injury in lung tissues was reduced,PaCO2 value,ratio of lung W/D,white blood cell count and eosinophils count in BALF,pathological injury score,MPO activity in lung tissues,IL-6 and COX-2 levels in BALF and serum,and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissues were all decreased,and PaO2 value was increased,the differences being statistically significant(P<0.05),and the improvement effect was more stronger in high-dose emodin group;coumermycin attenuated the improvement effect of emodin on the above various indexes in the model rats.Conclusion Emodin can reduce the inflammation levels in SAP rats through inhibiting the activation of JAK2/STAT3 signaling pathway,thus reducing the lung tissue damage and improving the lung function of rats.

OBJECTIVE: To explore and quantify the association of hot night exposure during the sperm development period (0-90 lag days) with semen quality. METHODS: A total of 6,640 male sperm donors from 6 human sperm banks in China during 2014-2020 were recruited in this multicenter study. Two indices (i.e., hot night excess [HNE] and hot night duration [HND]) were used to estimate the heat intensity and duration during nighttime. Linear mixed models were used to examine the association between hot nights and semen quality parameters. RESULTS: The exposure-response relationship revealed that HNE and HND during 0-90 days before semen collection had a significantly inverse association with sperm motility. Specifically, a 1 °C increase in HNE was associated with decreased sperm progressive motility of 0.0090 (95% confidence interval [ CI]: -0.0147, -0.0033) and decreased total motility of 0.0094 (95% CI: -0.0160, -0.0029). HND was significantly associated with reduced sperm progressive motility and total motility of 0.0021 (95% CI: -0.0040, -0.0003) and 0.0023 (95% CI: -0.0043, -0.0002), respectively. Consistent results were observed at different temperature thresholds on hot nights. CONCLUSION Our findings highlight the need to mitigate nocturnal heat exposure during spermatogenesis to maintain optimal semen quality.
Humans ; Male ; Semen Analysis ; Adult ; Sperm Motility ; Hot Temperature/adverse effects* ; China ; Middle Aged ; Spermatozoa/physiology* ; Young Adult

Objective:To evaluate the efficacy of peritoneal dialysis (PD) in the treatment of acute kidney injury (AKI) in critically ill neonates.Methods:It was a retrospective study. The baseline characteristic data, PD protocols, PD catheter placement methods and clinical outcomes of AKI neonates who underwent PD in the General Hospital of Ningxia Medical University between July 2015 and December 2024 were collected and analyzed.Results:(1) Among the 8 neonates with AKI, gestational age was (30.38±6.02) weeks, and birth weight was 1 397.5 (839.0, 2 312.5) g, with 6 premature infants. The time from birth to AKI onset was 144 (48, 294) hours. The leading cause of AKI was sepsis (6/8). The treatment time of PD was (93.12±37.20) hours. (2) Renal function recovery: After PD treatment, urine output was significantly increased ( Z=-3.29, P<0.001), and serum creatinine was significantly decreased ( t=2.66, P=0.032). (3) Hyperkalemia: Six out of 8 patients presented with hyperkalemia, which significantly decreased after PD treatment ( t=3.37, P=0.008). (4) Acid-base balance:Five out of 8 neonates had metabolic acidosis, and 3 of 5 neonates achieved basically complete correction (including lactic acidosis). There was no statistically significant difference in acid-base balance indicators before and after PD treatment (all P>0.05). (5) PD-related complications: Two out of 8 patients experienced peritoneal dialysate leakage, and no other PD-related complications occurred. (6) Outcomes: The hospital stay was 27.0 (8.0, 57.5) days. Four out of 8 neonates survived, while the other 4 neonates died after withdrawal of treatment. The primary cause was multiple organ failure. Conclusions:PD is a safe and effective treatment for neonatal AKI, facilitating early renal recovery and correction of electrolyte and acid-base imbalances.

Objective:To explore the impact of the Yiqi Huoxue formula combined with nucleos(t)ide analogs (NAs) antiviral therapy on the serum N-glycan abundance in patients with hepatitis B-related liver fibrosis so as to clarify the application value of the oligosaccharide chain test (GT) for dynamic monitoring of the liver fibrosis progression.Methods:Sixty-two cases diagnosed with chronic hepatitis B at the Department of Hepatology, the Third Hospital of Hebei Medical University, between January 2020 and December 2022 were enrolled and divided into a Yiqi Huoxue Formula (YQHX) combined with NAs group and an NAs monotherapy group ( n=31), with 31 cases in each group for a total of 96 weeks of follow-up. Patient's basic clinical characteristics and liver stiffness measurement (LSM) were collected. GT was used simultaneously to detect the serum N-glycan profile and abundance changes. The nonparametric Mann-Whitney U test was used to compare the differences between the two groups. Enumeration data were expressed as number of cases and percentages (%). The χ2 test was used to compare constituent ratios between two or more groups. Correlation analysis was performed using the Spearman method, with P<0.05 considered statistically significant. Results:The proportion of patients was significantly higher in the YQHX combined with NAs group than the NAs monotherapy group [61.29% (19/31) vs. 9.68% (3/31), P<0.05] with no progression in liver fibrosis staging following 96 weeks of follow-up. The abundance of the N-glycan marker peak 8 [triantennary N-glycan (NA3)] had resulted in significant change for liver fibrosis improvements ( P<0.05), which predicts liver fibrosis progression and reversal in populations sensitive to traditional Chinese medicine. Conclusion:The combined application of Yiqi Huoxue formula and NAs can significantly promote the improvement rate of hepatitis B-related liver fibrosis. Serum N-glycan peak 8 may serve as a potential biomarker for monitoring the reversal of hepatitis B-related liver fibrosis.

Objective:To analyze the association of apolipoprotein E(ApoE),soluble semaphoring 4D(sSema4D),and cyclophilin A(CypA)levels with unfavorable outcome in patients with chronic heart failure(CHF).Methods:We retrospectively enrolled 108 patients with CHF admitted in the Fifth Affiliated Hospital of Zhengzhou Universi-ty between March 2019 and March 2022.According to the outcome during 6-month follow-up,patients were di-vided into favorable outcome group(n=71)and unfavorable outcome group(n=37).ApoE,sSema4D and CypA levels were compared between the two groups,and multivariate Logistic regression was employed to identify the risk factors of unfavorable outcome within 6-month follow-up in CHF patients.Results:Compared to patients in the favorable outcome group,those in the unfavorable outcome group had significant higher proportion of NYHA classⅣ(59.46％vs.25.35％),serum creatinine[(80.74±3.89)μmol/L vs.(71.36±3.63)μmol/L],ApoE[(69.87±4.25)mg/L vs.(47.36±3.17)mg/L],sSema4D[(916.62±7.32)ng/L vs.(426.42±6.25)ng/L]and CypA[(6.74±1.32)ng/L vs.(4.38±0.72)ng/L](P＜0.001 all).Multivariate Logistic regression indicated that NY-HA class Ⅳ(OR=2.782,95％CI 1.166～6.636),serum creatinine(OR=29.893,95％CI 6.782～131.768),ApoE(OR=12.046,95％CI 4.694～30.913),sSema4D(OR=8.390,95％CI 2.260～31.146)and CypA(OR=6.486,95％CI 1.780～23.635)were independent risk factors for unfavorable outcome within 6-month follow-up in CHF patients(P＜0.05 or＜0.01).Conclusion:The levels of ApoE,sSema4D and CypA were closely associated with short-term unfavorable outcome in patients with chronic heart failure.