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*

Objective To elucidate the effect of Deduhonghua-7 powder and its main components on liver fibrosis induced by carbon tetrachloride(CCl4)by regulating chemokine receptor 1(CCR1)and DNA methyltransferase 1(DNMT1).Methods C57BL/6 mice were randomly divided into blank group,model group,Deduhonghua-7 powder group(0.75 g·kg-1 Deduhonghua-7 powder),Scabiosa Atropurea group(0.75 g·kg-1 Scabiosa Atropurea)and Luteolin-L,-H groups(0.02 and 0.04 g·kg-1 Luteolin,respectively).Except the blank group,the other groups were intrabitoneally injected with 10％CC14 olive oil solution to establish liver fibrosis model,and all groups were given 0.5％CMC-Na dissolved intragastric drug once a day for 4 weeks,and blood and liver tissues were collected after the last administration.The serum CCR1,DNMT1,α-smooth muscle actin(α-smA)and precollagen Ⅲ contents were detected by enzyme linked immunosorbent assay;the CCR1,DNMT1,α-smA,Collagen Ⅰprotein expression were detected by Western blot.Result The contents of serum DNMT1 in blank group,model group,Deduhonghua-7 powder group,Luteotin-L,-H groups and Scabiosa Atropurea group were(4.56±0.69),(6.09±0.59),(5.21±0.33),(4.99±0.68),(5.03±0.45)and(5.17±0.61)pg·mL-1;the contents of CCR1 were(13.38±0.47),(11.20±0.73),(12.97±0.80),(12.89±0.75)(12.88±0.95)and(12.92±0.58)pg·mL-1;the contents of α-smA were(181.80±24.50),(281.30±26.60),(220.90±22.30),(193.70±16.10),(199.30±17.70)and(205.80±14.70)pg·mL-1;the contents of procollagen Ⅲ were(49.29±8.26),(77.56±8.61),(67.56±5.63),(55.47±7.07),(64.93±8.66)and(59.66±8.51)pg·mL-1;the relative expression levels of DNMT1 protein in liver tissues were 0.08±0.03,0.26±0.08,0.13±0.01,0.12±0.05,0.13±0.05 and 0.15±0.03;the CCR1 protein relative expression levels were 0.18±0.03,0.13±0.02,0.21±0.06,0.22±0.07,0.17±0.07 and 0.18±0.06;the α-smA protein relative expression levels were 0.03±0.01,0.27±0.11,0.09±0.05,0.12±0.08,0.09±0.07 and 0.11±0.07;the expression of Colagen Ⅰ protein were 0.09±0.04,0.65±0.22,0.28±0.12,0.26±0.19,0.30±0.22 and 0.25±0.12.The differences of above indicators between the model group and the blank group,between the Deduhonghua-7 powder group,Luteotin-L,-H groups,Scabiosa Atropurea group and the model group were statistically significant(all P＜0.05).Conclusion Luteotein is one of the pharmacodynamic substance bases of Deduhonghua-7 powder to alleviate liver fibrosis induced by CC14.It can inhibit or delay the formation of liver fibrosis through CCR1,DNMT1,α-smA and Collagen Ⅰ proteins.

Objective To investigate the toxicokinetic differences of 3,4-methylenedioxy-N-methylamphetamine(MDMA)and its metabolite 4,5-methylene dioxy amphetamine(MDA)in rats af-ter single and continuous administration of MDMA,providing reference data for the forensic identifica-tion of MDMA.Methods A total of 24 rats in the single administration group were randomly divided into 5,10 and 20 mg/kg experimental groups and the control group,with 6 rats in each group.The ex-perimental group was given intraperitoneal injection of MDMA,and the control group was given intraperi-toneal injection of the same volume of normal saline as the experimental group.The amount of 0.5 mL blood was collected from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.In the continuous administration group,24 rats were randomly divided into the experi-mental group(18 rats)and the control group(6 rats).The experimental group was given MDMA 7 d by continuous intraperitoneal injection in increments of 5,7,9,11,13,15,17 mg/kg per day,respectively,while the control group was given the same volume of normal saline as the experimental group by in-traperitoneal injection.On the eighth day,the experimental rats were randomly divided into 5,10 and 20 mg/kg dose groups,with 6 rats in each group.MDMA was injected intraperitoneally,and the con-trol group was injected intraperitoneally with the same volume of normal saline as the experimental group.On the eighth day,0.5 mL of blood was taken from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.Liquid chromatography-triple quadrupole tandem mass spectrometry was used to detect MDMA and MDA levels,and statistical software was employed for data analysis.Results In the single-administration group,peak concentrations of MDMA and MDA were reached at 5 min and 1 h after administration,respectively,with the largest detection time limit of 12 h.In the continuous administration group,peak concentrations were reached at 30 min and 1.5 h af-ter administration,respectively,with the largest detection time limit of 10 h.Nonlinear fitting equations for the concentration ratio of MDMA and MDA in plasma and administration time in the single-administration group and continuous administration group were as follows:T=10.362C-1.183,R2=0.974 6;T=7.397 3C-0.694,R2=0.961 5(T:injection time;C:concentration ratio of MDMA to MDA in plasma).Conclusions The toxicokinetic data of MDMA and its metabolite MDA in rats,obtained through single and continuous administration,including peak concentration,peak time,detection time limit,and the relationship between concentration ratio and administration time,provide a theoretical and data foundation for relevant forensic identification.

Objective To investigate the independent risk factors of postoperative femoral head necrosis for pediatric femoral neck fractures.Methods The clinical data of 122 children with femoral neck fracture who underwent surgery in our hospital from July 2008 to July 2021 were retrospectively analyzed.The femoral head necrosis was counted,and the children were divided into the femoral head necrosis group and the femoral head normal group according to the presence of femoral head necrosis,then the risk factors of femoral head necrosis were analyzed.Results Of the 122 children,22 cases(18.03％)had femoral head necrosis.The postoperative femoral head necrosis for pediatric femoral neck fractures may be related to the age,Delbet classification,initial displacement degree,injury mechanism,operation time,internal fixation method and reduction quality(P＜0.05),while which has no significant correlation with the gender,side,time from injury to operation,or reduction method(P＞0.05).The results of Logistic regression analysis showed that age＞10 years,Delbet type of Ⅰ to Ⅱ,initial displacement degree of type Ⅲ,high-energy injury and unacceptable reduction were the independent risk factors of postoperative femoral head necrosis for pediatric femoral neck fractures(P＜0.05).Conclusion Older children,or patients with Delbet type of Ⅰ to Ⅱ,large degree of initial displacement,high-energy injury,and unacceptable reduction are the independent risk factors of postoperative femoral head necrosis for pediatric femoral neck fractures.

Mitral annular calcification(MAC)is an age-related,chronic,degenerative change in localized fibrous support structures,and current research suggests that it is a process similar to the active onset of atherosclerosis and aortic valve calcification,both of which are accompanied by the deposition of lipoprotein(α)[Lp(α)]and the formation of chronic inflammatory foci.Among them,Lp(α)is the hot spot of research.In recent years,the relationship between Lp(α)and aortic valve calcification has received extensive attention.A large number of studies have demonstrated that elevated Lp(α)and its associated oxidized phospholipids(OxPL)can promote aortic valve calcification and stenosis through multiple calcium-regulated pathways,but the pathophysiological process of MAC is much more complex and unclear,and there has been a preliminary exploration of the relationship between Lp(α)and MAC.To make the current relationship between the two clearer,and thus provide new possibilities for preventing or delaying MAC,the paper will review the three aspects of MAC,Lp(α),and the research progress between the two.

The detailed knowledge of the morphological structure, drainage pathways and patterns, the first tier lymph node of the cardiac lymphatic and its relationship with the circulatory system has not yet been completed. Although, the cardiac lymphatics had been described with renewed interest in past years, which was attributed to the transparent nature of lymphatic vessels that are difficult to be observed. In this study, cardiac lymphatics of the goat heart were perfused by a direct microinjecting technique with a radiopaque mixture. This demonstrated the subepicardial and subendocardial lymph capillary networks communicating with transmyocardial lymph vessels and then entering to subepicardial collecting lymph vessels that were directed toward the atrio-ventricular sulcus where they form a confluence from which the main cardiac lymph channels. We also found that: 1) the quantity and caliber of collecting lymph vessels varied in each goat heart; 2) drainage patterns of lymph vessels in the goat heart were different in individuals; 3) the first tier lymph node that each major lymph vessel drained to was different; and 4) multiple lymphatic-venous anastomosis sites have been confirmed to exist in the subepicardium of the left and right ventricles of each goat heart, which may be the morphological structure to accelerate the return of intercellular fluid to the venous system during excessive exercise of the heart. Therefore, the information may provide reference for further study in physiological and pathological conditions of the human heart.

The detailed knowledge of the morphological structure, drainage pathways and patterns, the first tier lymph node of the cardiac lymphatic and its relationship with the circulatory system has not yet been completed. Although, the cardiac lymphatics had been described with renewed interest in past years, which was attributed to the transparent nature of lymphatic vessels that are difficult to be observed. In this study, cardiac lymphatics of the goat heart were perfused by a direct microinjecting technique with a radiopaque mixture. This demonstrated the subepicardial and subendocardial lymph capillary networks communicating with transmyocardial lymph vessels and then entering to subepicardial collecting lymph vessels that were directed toward the atrio-ventricular sulcus where they form a confluence from which the main cardiac lymph channels. We also found that: 1) the quantity and caliber of collecting lymph vessels varied in each goat heart; 2) drainage patterns of lymph vessels in the goat heart were different in individuals; 3) the first tier lymph node that each major lymph vessel drained to was different; and 4) multiple lymphatic-venous anastomosis sites have been confirmed to exist in the subepicardium of the left and right ventricles of each goat heart, which may be the morphological structure to accelerate the return of intercellular fluid to the venous system during excessive exercise of the heart. Therefore, the information may provide reference for further study in physiological and pathological conditions of the human heart.

The detailed knowledge of the morphological structure, drainage pathways and patterns, the first tier lymph node of the cardiac lymphatic and its relationship with the circulatory system has not yet been completed. Although, the cardiac lymphatics had been described with renewed interest in past years, which was attributed to the transparent nature of lymphatic vessels that are difficult to be observed. In this study, cardiac lymphatics of the goat heart were perfused by a direct microinjecting technique with a radiopaque mixture. This demonstrated the subepicardial and subendocardial lymph capillary networks communicating with transmyocardial lymph vessels and then entering to subepicardial collecting lymph vessels that were directed toward the atrio-ventricular sulcus where they form a confluence from which the main cardiac lymph channels. We also found that: 1) the quantity and caliber of collecting lymph vessels varied in each goat heart; 2) drainage patterns of lymph vessels in the goat heart were different in individuals; 3) the first tier lymph node that each major lymph vessel drained to was different; and 4) multiple lymphatic-venous anastomosis sites have been confirmed to exist in the subepicardium of the left and right ventricles of each goat heart, which may be the morphological structure to accelerate the return of intercellular fluid to the venous system during excessive exercise of the heart. Therefore, the information may provide reference for further study in physiological and pathological conditions of the human heart.

The detailed knowledge of the morphological structure, drainage pathways and patterns, the first tier lymph node of the cardiac lymphatic and its relationship with the circulatory system has not yet been completed. Although, the cardiac lymphatics had been described with renewed interest in past years, which was attributed to the transparent nature of lymphatic vessels that are difficult to be observed. In this study, cardiac lymphatics of the goat heart were perfused by a direct microinjecting technique with a radiopaque mixture. This demonstrated the subepicardial and subendocardial lymph capillary networks communicating with transmyocardial lymph vessels and then entering to subepicardial collecting lymph vessels that were directed toward the atrio-ventricular sulcus where they form a confluence from which the main cardiac lymph channels. We also found that: 1) the quantity and caliber of collecting lymph vessels varied in each goat heart; 2) drainage patterns of lymph vessels in the goat heart were different in individuals; 3) the first tier lymph node that each major lymph vessel drained to was different; and 4) multiple lymphatic-venous anastomosis sites have been confirmed to exist in the subepicardium of the left and right ventricles of each goat heart, which may be the morphological structure to accelerate the return of intercellular fluid to the venous system during excessive exercise of the heart. Therefore, the information may provide reference for further study in physiological and pathological conditions of the human heart.