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Abstract Extracellular vesicles (EVs) are highly heterogeneous, and different EV subpopulations from various origins mediate different biological effects. The separation of different subpopulations of EVs from mixtures is critical but challenging. Epididymosomes are secreted by the epididymal epithelium and play a key role in sperm maturation and function. However, limited access to human epididymal tissue and epididymal fluid has hampered further study of epididymosomes and their potential clinical applications. Here, we established a novel strategy based on flow cytometry sorting to isolate human CD63‐positive epididymosomes from ejaculate. We identified CD52, a membrane‐located protein expressed exclusively in the epididymis, as the sorting marker for human epididymosomes. Then, CD63‐positive epididymosomes were isolated from human semen using a flow cytometry sorting instrument and concentrated. Additionally, we observed that isolated CD63‐positive epididymosomes improved sperm function more than other CD63‐positive seminal EV subpopulations did, demonstrating the successful isolation of a subpopulation of epididymosomes from human semen and their potential application in the clinic.

Abstract Background Small extracellular vesicles (EVs), exemplified by exosomes, mediate intercellular communication by transporting proteins, mRNAs, and miRNAs. Post-translational modifications are involved in controlling small EV secretion process. However, whether palmitoylation regulates small EV secretion, remains largely unexplored. Methods Vacuole Membrane Protein 1 (VMP1) was testified to be S-palmitoylated by Palmitoylation assays. VMP1 mutant plasmids were constructed to screen out the exact palmitoylation sites. Small EVs were isolated, identified and compared between wild-type VMP1 or mutant VMP1 transfected cells. Electron microscope and immunofluorescence were used to detect multivesicular body (MVB) number and morphology change when VMP1 was mutated. Immunoprecipitation and Mass spectrum were adopted to identify the protein that interacted with palmitoylated VMP1, while knock down experiment was used to explore the function of targeted protein ALIX. Taking human Sertoli cells (SCs) and human spermatogonial stem cell like cells (SSCLCs) as a model of intercellular communication, SSCLC maintenance was detected by flow cytometry and qPCR at 12 days of differentiation. In vivo, mouse model was established by intraperitoneal injection with palmitoylation inhibitor, 2-bromopalmitate (2BP) for 3 months. Results VMP1 was identified to be palmitoylated at cysteine 263,278 by ZDHHC3. Specifically, palmitoylation of VMP1 regulated its subcellular location and enhanced the amount of small EV secretion. Mutation of VMP1 palmitoylation sites interfered with the morphology and biogenesis of MVBs through suppressing intraluminal vesicle formation. Furthermore, inhibition of VMP1 palmitoylation impeded small EV secretion by affecting the interaction of VMP1 with ALIX, an accessory protein of the ESCRT machinery. Taking SCs and SSCLCs as a model of intercellular communication, we discovered VMP1 palmitoylation in SCs was vital to the growth status of SSCLCs in a co-culture system. Inhibition of VMP1 palmitoylation caused low self-maintenance, increased apoptosis, and decreased proliferation rate of SSCLCs. In vivo, intraperitoneal injection of 2BP inhibited VMP1 palmitoylation and exosomal marker expression in mouse testes, which were closely associated with the level of spermatogenic cell apoptosis and proliferation. Conclusions Our study revealed a novel mechanism for small EV secretion regulated by VMP1 palmitoylation in Sertoli cells, and demonstrated its pivotal role in intercellular communication and SSC niche.

Abstract Background Dietary patterns play an important role in regulating serum uric acid levels in the body, but evidence for the association between different kinds of plant-based and animal-based dietary patterns and individual serum uric acid levels is scarce and inconsistent. Methods We analyzed data from the sixth wave of the China Health and Nutrition Survey. The plant-based diet of 7,806 participants was determined using three consecutive 24-hour dietary recalls, and latent profile analysis was used to identify dietary patterns among participants. Serum uric acid levels were analyzed using the enzymatic colorimetric method. The association between intakes of different types of dietary pattern and individual serum uric acid levels was analyzed using linear regression analysis, after adjusting for confounding variables. Results We identified three types of plant-based dietary patterns, namely, low tuber starches and vegetable plant-based diet (LTVP), high cereal, tuber starches and vegetable plant-based diet (HCTVP), and high legume and fruit plant-based diet (HLFP). We also identified three types of animal-based dietary patterns, namely, high milk and egg animal-based diet (HMiEA), low egg and fish animal-based diet, and high meat and fish animal-based diet (HMeFA). Significant coefficients for participant serum uric acid levels were observed for the HCTVP diet (β = −0.022, P = 0.031) and HMeFA diet (β = 0.061, P

Groundwater in mining areas play an important supporting role in the surrounding residents' living and the construction of coal industries. Mine drainage during coal mining will change the original groundwater circulation process in mining areas, leading to changes in the groundwater environment. Therefore, clarifying the mutual transformation relationship between the “three waters” is the key to the scientific utilization of water resources in mining areas. Chemical components and hydrogen and oxygen isotopes of the groundwater in the Pingyu mining area are combined to determine the characteristics of groundwater circulation after large-scale drainage of the mine, identify the origin and controlling factors of the major components in the groundwater. Hydrohemical analyses show that most of the groundwater are of HCO3—Ca·Mg type, The hydrochemical components are controlled by the weathering and dissolution of silicate, and the ions in the groundwater mainly come from water-rock interactions and the waste generated by human activities, with concentration of \begin{document}${\rm{NO}}_3^- $\end{document} exceeding the standard. The hydrogen and oxygen isotopic data demonstrates that the groundwater in the study area receives recharge from modern precipitation, and there is a close hydraulic relationship between the groundwater in the unconsolidated aquifers and the groundwater in the karst aquifer. After the mine drainage, the groundwater circulation process in the mining area changes, the karst groundwater changes from supporting the upper pore groundwater to accepting the leakage recharge of the upper pore groundwater, and then discharges to the surface water by the mine drainage. The mine drainage becomes the main groundwater drainage way in the mine area. The research results can provide scientific and effective basis for groundwater development management and land subsidence prevention in mining areas.

ABSTRACTChimeric antigen receptor (CAR) T cell immunotherapy has demonstrated success in the treatment of hematological malignancies; however, its efficacy and applications in solid tumors remain limited. Immunosuppressive factors, particularly inhibitory checkpoint molecules, restrict CAR T cell activity inside solid tumors. The modulation of checkpoint pathways has emerged as a promising approach to promote anti-tumor responses in CAR T cells. Programmed cell death protein 1 (PD1) and T cell immunoreceptor with Ig and ITIM domains (TIGIT) are two critical immune-checkpoint molecules that suppress anti-tumor activity in T cells. Simultaneous targeting of these two inhibitory molecules could be an efficient checkpoint modulation strategy. Here, we developed a PD1-TIGIT chimeric immune-checkpoint switch receptor (CISR) that enhances the efficacy of CAR T cell immunotherapy by reversing the inhibitory checkpoint signals of PD1/PDL1 and/or TIGIT/CD155. In addition to neutralizing PDL1 and CD155, this chimeric receptor is engineered with the transmembrane region and intracellular domain of CD28, thereby effectively enhancing T cell survival and tumor-targeting functions. Notably, under simultaneous stimulation of PDL1 and CD155, CISR-CAR T cells demonstrate superior performance in terms of cell survival, proliferation, cytokine release, and cytotoxicity in vitro, compared with conventional CAR T cells. Experiments utilizing both cell line- and patient-derived xenotransplantation tumor models showed that CISR-CAR T cells exhibit robust infiltration and anti-tumor efficiency in vivo. Our results highlight the potential for the CISR strategy to enhance T cell anti-tumor efficacy and provide an alternative approach for T cell-based immunotherapies.

The surface modification of aramid fiber plain fabric (PPTA) was conducted through phthalic anhydride treatment and anhydrous aluminum chloride (AlCl3) catalysis, aiming to enhance the interfacial bonding strength between aramid fiber fabric and bisphenol A diglycidyl ether (DGEBA) resin. The surface morphologies and structures of PPTA fiber before and after modification were characterized using scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and X-ray diffractometry. The mechanical properties of the PPTA/DGEBA composite were evaluated using a universal mechanical testing machine. The results demonstrate that when the concentration of phthalic anhydride is 0.3 mol/L, the tensile strength, bending strength and interlaminar shear strength of PPTA/DGEBA composites reach the maximum value, which are increased by 17.94%, 44.18%, and 15.94% compared with the unmodified sample, respectively. After a 0.5-h catalytic modification, the PPTA/DGEBA composites exhibited significantly enhanced tensile strength, bending strength, and interlaminar shear strength, achieving respective increments of 32.28%, 24.91%, and 29.10% compared to the modified samples without catalyst addition. Moreover, the overall mechanical properties of the aramid fiber fabrics and composites were substantially improved, which are more suitable for structural applications.

Introduction: Metal corrosion detection is important for protecting lives and property. X-ray inspection systems are widely used because of their good penetrability and visual presentation capability. They can visually display both external and internal corrosion defects. However, existing X-ray-based defect detection methods cannot present and estimate the dense corrosion depths. To solve this problem, we propose a dense metal corrosion depth estimation method based on image segmentation and inpainting.Methods: The proposed method employs an image segmentation module to segment metal corrosion defects and an image inpainting module to remove these segmented defects. It then calculates the pixel-level dense corrosion depths using the X-ray images before and after inpainting. Moreover, to address the difficulty of acquiring training images with ground-truth dense corrosion depth annotations, we propose a virtual data generation method for creating virtual corroded metal X-ray images and their corresponding ground-truth annotations.Results: Experiments on both virtual and real datasets show that the proposed method successfully achieves accurate dense metal corrosion depth estimation.Discussion: In conclusion, the proposed virtual data generation method can provide effective and sufficient training samples, and the proposed dense metal corrosion depth estimation framework can produce accurate dense corrosion depths.

Introduction: The presence of connected and automated vehicles (CAV) in mixed traffic flows with different market penetration rates (MPRs) in urban road scenarios has a significant effect on fuel consumption and exhaust emissions.Methods: Therefore, in this study, real-world road networks and traffic data are simulated using SUMO based on actual data from a survey. The fuel consumption and emission benefits of CAVs in mixed traffic flows are well-evaluated, and the energy-saving performance of CAVs under low-speed vehicle interference is tested. In addition, this study explores both the energy consumption and emissions of purely electric vehicles.Results: The results show that with 100% CAV penetration, fuel vehicles have a maximum reduction in fuel consumption of 18% and a maximum increase in average speed of 31.6%, while the energy consumption of electric vehicles increases due to communication, detection, and collaboration between CAVs.Discussion: However, the results clearly demonstrate that the carbon emissions of electric vehicles are significantly lower than fuel vehicles. In addition, the increase in low-speed vehicles will result in an increase in energy consumption and emissions. Therefore, increasing the percentage of electric vehicles on the roads and transitioning from manual to autonomous driving systems is crucial to curbing carbon emissions.

Abstract Background Sperm, during epididymal transit, acquires microRNAs(miRNAs), which are crucial for embryonic development. However, whether sperm miRNAs influenced by an obstructive epididymal environment affect embryonic development remains unknown. Method The sham operation and vasectomy were performed in C57BL/6 J mice to create the control group (CON) and the obstructive epididymal environment group(OEE) group, respectively. The morphology of the testis and epididymis was observed using hematoxylin and eosin staining (HE staining) to establish the OEE mice model. The sperm quality test, intracytoplasmic sperm injection (ICSI), and epididymosomes fusion were employed to observe the effect of the obstructive epididymal environment on sperm and resultant embryonic development. The alteration of the sperm small RNA (sRNA) profile was analyzed by sRNA sequencing. RT-qPCR and DNA methylation were applied to observe the effect of obstructive epididymis on the expression of sperm miRNAs. The miRNAs microinjection was used to explore the impacts of sperm miRNAs on embryonic development. Results We confirmed postoperative 8-week mice as the OEE mice model by examining the morphology of the testis and epididymis. In the OEE group, we observed that sperm quality degraded and the development potential of embryos was reduced, which can be saved by the normal epididymal environment. The sperm sRNA sequencing revealed that the expression of the developmental miR-17–92 cluster and the Sfmbt2 miRNA cluster was downregulated in the OEE group. The expression of these two miRNA clusters in epididymis was also downregulated and regulated by DNA methylation. However, the downregulation of either the miR-17–92 cluster or the Sfmbt2 miRNA cluster in normal zygotes did not impair embryonic development. Conclusion The obstructive epididymal environment influences sperm quality and resultant embryonic development, as well as the abundance of the developmental miR-17–92 cluster and the Sfmbt2 miRNA cluster in sperm, but these miRNA clusters are not the cause of abnormal embryonic development. It implies that epididymis is important in early embryonic development and may play a potential role in sperm epigenome.

Abstract Background Human primordial germ cells (hPGCs) initiate from the early post-implantation embryo at week 2–3 and undergo epigenetic reprogramming during development. However, the regulatory mechanism of DNA methylation during hPGC specification is still largely unknown due to the difficulties in analyzing early human embryos. Using an in vitro model of hPGC induction, we found a novel function of TET proteins and NANOG in the hPGC specification which was different from that discovered in mice. Methods Using the CRISPR–Cas9 system, we generated a set of TET1, TET2 and TET3 knockout H1 human embryonic stem cell (hESC) lines bearing a BLIMP1-2A-mKate2 reporter. We determined the global mRNA transcription and DNA methylation profiles of pluripotent cells and induced hPGC-like cells (hPGCLCs) by RNA-seq and whole-genome bisulfite sequencing (WGBS) to reveal the involved signaling pathways after TET proteins knockout. ChIP-qPCR was performed to verify the binding of TET and NANOG proteins in the SOX17 promoter. Real-time quantitative PCR, western blot and immunofluorescence were performed to measure gene expression at mRNA and protein levels. The efficiency of hPGC induction was evaluated by FACS. Results In humans, TET1, TET2 and TET3 triple-knockout (TKO) human embryonic stem cells (hESCs) impaired the NODAL signaling pathway and impeded hPGC specification in vitro, while the hyperactivated NODAL signaling pathway led to gastrulation failure when Tet proteins were inactivated in mouse. Specifically, TET proteins stimulated SOX17 through the NODAL signaling pathway and directly regulates NANOG expression at the onset of hPGCLCs induction. Notably, NANOG could bind to SOX17 promoter to regulate its expression in hPGCLCs specification. Furthermore, in TKO hESCs, DNMT3B-mediated hypermethylation of the NODAL signaling-related genes and NANOG/SOX17 promoters repressed their activation and inhibited hPGCLC induction. Knockout of DNMT3B in TKO hESCs partially restored NODAL signaling and NANOG/SOX17 expression, and rescued hPGCLC induction. Conclusion Our results show that TETs-mediated oxidation of 5-methylcytosine modulates the NODAL signaling pathway and its downstream genes, NANOG and SOX17, by promoting demethylation in opposition to DNMT3B-mediated methylation, suggesting that the epigenetic balance of DNA methylation and demethylation in key genes plays a fundamental role in early hPGC specification.

Abstract Background Although association of depressive symptoms with cigarette or alcohol is well documented, the dose–response relationship between them is rarely studied. This study aims to evaluate dose–response relationships of cigarette and alcohol consumption with depressive symptoms in Chinese middle-aged and elderly men, providing evidence to guide cigarette and alcohol control. Methods This multiple-center, cross-sectional study including 5965 Chinese men aged 40–79 years was conducted in 2013–2016 in China. Depressive symptoms were evaluated by Beck Depression Inventory-Short Form. History of cigarette smoking and alcohol drinking were collected with a structured questionnaire. Prevalence of depressive symptoms was compared depending on cigarette and alcohol consumption. Adjusted odds ratios (OR) and 95% confidence interval (CI) were estimated by binary logistic regression. Interpolation analysis was applied to test dose–effect relationships. Results A parabolic-shaped relationship was observed between cigarette consumption and depressive symptoms. Compared to never smokers, 59.0% (OR = 1.59, 95% CI 1.30–1.94) and 29.0% (OR = 1.29, 95% CI 1.08–1.54) higher odds of depressive symptoms were observed in men smoking 20 cigarettes/day (P = 0.092). An inverted J-shaped relationship was observed between alcohol consumption and depressive symptoms. Compared to never drinkers, a tendency of higher prevalence of depressive symptoms (OR = 1.16, 95% CI 0.99–1.36) was observed in men drinking 280 g/week. Conclusions Associations of cigarette smoking and alcohol drinking with depressive symptoms differ with consumption in middle-aged and elderly men. Health-care providers should exercise great caution on depressive symptoms in conducting cigarette and alcohol control.

Abstract Background The most serious condition of male infertility is complete Sertoli cell-only syndrome (SCOS), which refers to the lack of all spermatogenic cells in the testes. The genetic cause of SCOS remains to be explored. We aimed to investigate the genetic cause of SCOS and assess the effects of the identified causative variant on human male germ cells. Methods Whole-exome sequencing was performed to identify potentially pathogenic variants in a man with complete SCOS, and Sanger sequencing was performed to verify the causative variant in this man and his father and brother. The pathogenic mechanisms of the causative variant were investigated by in vitro differentiation of human-induced pluripotent stem cells (hiPSCs) into germ cell-like cells. Results The homozygous loss-of-function (LoF) variant p.His244ArgfsTer31 (c.731_732delAT) in PIWIL2 was identified as the causative variant in the man with complete SCOS, and the same variant in heterozygosis was confirmed in his father and brother. This variant resulted in a truncated PIWIL2 protein lacking all functional domains, and no PIWIL2 expression was detected in the patient’s testes. The patient and PIWIL2 −/− hiPSCs could be differentiated into primordial germ cell-like cells and spermatogonial stem cell-like cells (SSCLCs) in vitro, but the formation and maintenance of SSCLCs were severely impaired. RNA-seq analyses suggested the inactivation of the Wnt signaling pathway in the process of SSCLC induction in the PIWIL2 −/− group, which was validated in the patient group by RT-qPCR. The Wnt signaling pathway inhibitor hindered the formation and maintenance of SSCLCs during the differentiation of normal hiPSCs. Conclusions Our study revealed the pivotal role of PIWIL2 in the formation and maintenance of human spermatogonial stem cells. We provided clinical and functional evidence that the LoF variant in PIWIL2 is a genetic cause of SCOS, which supported the potential role of PIWIL2 in genetic diagnosis. Furthermore, our results highlighted the applicability of in vitro differentiation models to function validation experiments.

Accurate indoor occupancy information extraction plays a crucial role in building energy conservation. Vision-based methods are popularly used for occupancy information extraction because of their high accuracy. However, previous vision-based methods either only provide 2D occupancy information or require expensive equipment. In this paper, we propose a cost-effective indoor occupancy information extraction system that estimates occupant positions and trajectories in 3D using a single RGB camera. The proposed system provides an inverse proportional model to estimate the distance between a human head and the camera according to pixel-heights of human heads, eliminating the dependence on expensive depth sensors. The 3D position coordinates of human heads are calculated based on the above model. The proposed system also associates the 3D position coordinates of human heads with human tracking results by assigning the 3D coordinates of human heads to the corresponding human IDs from a tracking module, obtaining the 3D trajectory of each person. Experimental results demonstrate that the proposed system successfully calculates accurate 3D positions and trajectories of indoor occupants with only one surveillance camera. In conclusion, the proposed system is a low-cost and high-accuracy indoor occupancy information extraction system that has high potential in reducing building energy consumption.

Off-axis parabolic mirrors have extensive applications in X-ray optics, with the precision of their curvature directly impacting grazing-incidence focusing performance. Notably, the off-axis parabolic surface has non-rotating and non-symmetrical characteristics. Ultra-precision raster grinding utilizing a diamond wheel is a common method. Crucially, establishing an optimal wheel path stands as the key to ensuring surface accuracy during off-axis paraboloid grinding. In this study, according to the double curvature property of an off-axis parabolic surface, two different wheel paths were compared: one tracing the meridian direction (parabolic generatrix) and the other following the arc vector direction (arc). The results showed that the wheel path in raster grinding stepping along the arc vector direction can obtain a smaller scallop height and higher surface accuracy. The surface accuracy of one step along the arc vector direction is 9.6 μm, and that of the other step along the meridian direction is 32.6 μm. A model of the scallop height was established based on the relative relationship between adjacent wheel paths, and the error is within 5%. According to the correlation between scallop height and shape error, we conducted an analysis of the spatial distribution of shape errors under varying wheel paths. The wheel path that steps along the arc vector is more suitable for raster grinding of the off-axis paraboloid. The above study can provide theoretical guidance for the wheel path planning of off-axis parabolic mirrors with high surface accuracy.

Abstract Background Studying human germ cell development and male infertility is heavily relied on mouse models. In vitro differentiation of human pluripotent stem cells into spermatogonial stem cell-like cells (SSCLCs) can be used as a model to study human germ cells and infertility. The current study aimed to develop the SSCLC induction protocol and assess the effects of the developed protocol on SSCLC induction. Methods We examined the effects of valproic acid (VPA), vitamin C (VC) and the combination of VPA and VC on the SSCLC induction efficiency and determined the expression of spermatogonial genes of differentiated cells. Haploid cells and cells expressed meiotic genes were also detected. RNA-seq analysis was performed to compare the transcriptome between cells at 0 and 12 days of differentiation and differently expressed genes were confirmed by RT-qPCR. We further evaluated the alteration in histone marks (H3K9ac and H3K27me3) at 12 days of differentiation. Moreover, the SSCLC induction efficiency of two hiPSC lines of non-obstructive azoospermia (NOA) patients was assessed using different induction protocols. Results The combination of low concentrations of VPA and VC in the induction medium was most effective to induce SSCLCs expressing several spermatogonial genes from human pluripotent stem cells at 12 days of differentiation. The high concentration of VPA was more effective to induce cells expressing meiotic genes and haploid cells. RNA-seq analysis revealed that the induction of SSCLC involved the upregulated genes in Wnt signaling pathway, and cells at 12 days of differentiation showed increased H3K9ac and decreased H3K27me3. Additionally, two hiPSC lines of NOA patients showed low SSCLC induction efficiency and decreased expression of genes in Wnt signaling pathway. Conclusions VPA robustly promoted the differentiation of human pluripotent stem cells into SSCLCs, which involved the upregulated genes in Wnt signaling pathway and epigenetic changes. hiPSCs from NOA patients showed decreased SSCLC induction efficiency and Wnt signaling pathway gene expression, suggesting that SSC depletion in azoospermia testes might be associated with inactivation of Wnt signaling pathway. Our developed SSCLC induction protocol provides a reliable tool and model to study human germ cell development and male infertility.

In order to improve the bearing capacity of a bridge under dynamic impact and at the same time to improve the durability and safety of the bridge, the bridge should be jacked up, and most of the current research on bridge jacking upgrading is on synchronous jacking upgrading. There has been little research on single-pier jacking upgrading, especially the dynamics response problem under single-pier jacking upgrading of the bridge. This paper is based on the principle of vehicle–bridge coupling, and the vehicle was analyzed at different speeds by using the established model, vehicle weight, and multi-lane driving, comparing the dynamic response of the bridge before and after the single-pier jacking retrofit. The study analyzed the impact of the bridge single-pier jacking retrofit on the bridge impact coefficient, and evaluated the impact coefficient of the bridge from the perspective of dynamics. A comprehensive evaluation of the bridge single-pier jacking retrofit project was carried out. The following conclusions are drawn: the single-pier jacking modification of the bridge increased the bridge’s capacity under dynamic impact and also enhanced the durability and safety of the bridge.

The traditional gauge wheel has poor performance in reducing the adhesion to soil and constructing seed furrow, which results in lower seeding quality of the planter. To reduce the adhesion of the gauge wheel to the soil and build a well-structured seed furrow, an elastic gauge wheel with soil retention groove and irregular cavity was designed in this study. The soil retention groove built ridges on both sides of the seed furrow and avoided the gauge wheel compacting the seed furrow sidewalls. The irregular cavity increased the elasticity of the gauge wheel and allowed the wheel to squeeze the soil on both sides of the seed furrow, which reduced the soil adhesion of the wheel and built stable ridges. Soil moisture content was chosen as the experimental factor for comparative tests to evaluate the soil adhesion and the constructed seed furrow of the gauge wheel with an irregular cavity and the traditional gauge wheel. The experimental results showed that the viscosity reduction rate of the gauge wheel with the irregular cavity was not less than 12.61%. Compared with the traditional gauge wheel, the seed furrow constructed by the irregular cavity gauge wheel had ridges on both sides and less backfill soil, and the soil compaction of sidewalls decreased by 18.16%. The field experiment was designed using the Box–Behnken design. The working speed, downforce, and planting depth were taken as experimental factors, and the soil adhesion of the gauge wheel and the consistency of planting depth were taken as evaluation indicators. The optimal operating parameters of planter obtained by Design-Expert 8.0.6 software were as follows: the working speed was 8 km·h−1, the downforce was 844 N, and the planting depth was 65 mm. The verification test of the optimal operating parameters showed that the soil adhesion mass of the gauge wheel was 123.65 g and the coefficient of variation of the planting depth was 5.35%. This study provides a reference for the mechanized construction method of seed furrow by precision planter and the structural design and performance optimization of gauge wheels.

Abstract The fertilizing spermatozoon is a highly specialized cell that selects from millions along the female tract until the oocyte. The paternal components influence the oocyte activation during fertilization and are fundamental for normal embryo development; however, the sperm-oocyte interplay is in a continuous debate. This review aims to analyze the available scientific information related to the role of the male gamete in the oocyte activation during fertilization, the process of the interaction of sperm factors with oocyte machinery, and the implications of any alterations in this interplay, as well as the advances and limitations of the reproductive techniques and diagnostic tests. At present, both PLCζ and PAWP are the main candidates as oocyte activated factors during fertilization. While PLCζ mechanism is via IP3, how PAWP activates the oocyte still no clear, and these findings are important to study and treat fertilization failure due to oocyte activation, especially when one of the causes is the deficiency of PLCζ in the sperm. However, no diagnostic test has been developed to establish the amount of PLCζ, the protocol to treat this type of pathologies is broad, including treatment with ionophores, sperm selection improvement, and microinjection with PLCζ protein or RNA.

Emerging viral infections continuously pose a threat to human wellbeing. Several RNA viruses have managed to establish access to the male reproductive tract and persist in human semen. The sexual transmission of the virus is of critical public concern. The epidemiological inferences are essential to understand its complexity, particularly the probability of viral transmission from asymptomatic patients or those in the incubation period or from the patient who was previously infected and now fully recovered. From the clinical perspective, negative impacts in the male reproductive tract associated with RNA virus infection have been described, including orchitis, epididymitis, impaired spermatogenesis, and a decrease in sperm quality, which can affect male fertility at different time intervals. The disruption of anatomical barriers due to inflammatory responses might enable the viral invasion into the testis, and the immune privilege status of testes might facilitate a sustained persistence of the virus in the semen. In this review, the current knowledge about other RNA viruses that affect male reproductive health provides the framework to discuss the impact of the SARS-CoV-2 pandemic. The molecular mechanisms, sexual transmission, and viral impacts for mumps, HIV, Zika, and Ebola viruses are explored. We discuss the currently available information on the impact of SARS-CoV-2 and its sequelae in the male reproductive tract, particularly regarding presence in semen, its impact on sexual organs, and sperm quality. To date, no sexual transmission of SARS-CoV-2 has been reported, whereas the identification of viral particles in semen remains conflicting. In the purview of the earlier conducted analyses, it is essential to investigate further the long-term health impacts of SARS-CoV-2 on the male reproductive tract.

Abstract Background As the precursors of sperm and eggs, human primordial germ cells (hPGCs) emerge as early as weeks 2 to 3 of post-implantation development. Recently, robust hPGC induction models have been established in vitro with different protocols, but global 5mC/5hmC epigenetic reprogramming is not initiated in vitro. Previous studies found that vitamin C can enhance Tet (ten-eleven translocation) enzyme expression and improve 5hmC level in cells. But the effect of vitamin C supplementation on hPGC in vitro induction is still unknown. Methods We generated a gene-edited human embryonic stem cell (hESC) line carrying a BLIMP1-mkate2 reporter by CRISPR/Cas9 technology and used flow cytometry to optimize the PGC differentiation protocol; meanwhile, the expression of PGC genes (BLIMP1, TFAP2C, SOX17, OCT4) was evaluated by qRT-PCR. When different concentrations of vitamin C were added to the induction medium, the percentage of hPGCLCs (hPGC-like cells) was analyzed by flow cytometry; dot blot and ELISA were used to detect the levels of 5hmC and 5mC. The expression of TET enzymes was also evaluated by qRT-PCR. Results We optimized the PGC differentiation protocol with the BLIMP1-mkate reporter hESCs, and the efficiency of PGC induction in vitro can be improved to 30~40%. When 50 μg/mL vitamin C was added, the derived hPGCLCs not only upregulated the expression of key genes involved in human early germ cell development such as NANOS3, TFAP2C, BLIMP1, and SOX17, but also increased the levels of 5hmC and TET enzymes. Conclusions Taken together, supplementation of vitamin C can promote the in vitro induction of hPGCLCs from hESCs, which might be related to vitamin C-mediated epigenetic regulations during the differentiation process.

Abstract Background Aberrant DNA damage of germ cells, which impairs spermatogenesis and lowers fertility, is an important factor contributing to male infertility. MicroRNAs (miRNAs) play a significant role in the expression and regulation of multiple genes during spermatogenesis. Our previous study found much lower miR-424 (murine homologue miR-322) levels in the seminal plasma of infertile patients with high DFI(DNA Fragmentation Index)than in the fertile group. However, the mechanism by which miR-322 regulates germ cells during spermatogenesis remains unknown. Methods In this study, we successfully established a GC-2 cell model of miR-322 downregulation resulting in impaired spermatogenesis. And the cell viability were measured using Cell Counting Kit-8 (CCK-8; Dojindo, Japan) and MTT (Sigma Aldrich, USA). Immunofluorescence assay was used to detect cell damage and the expression of apoptosis-related proteins were measured using real-time quantitative PCR and Western blot analysis. Target genes were predicted and verified by online database retrieval and Dual-luciferase reporter gene assay. Results We observed evident decreases in the cell viability of GC-2 cells along with remarkable increases in apoptosis after miR-322 inhibition. While the expression of apoptosis-related genes, including Bax and caspases 3, 9, and 8 greatly increased in GC-2 cells after miR-322 downregulation, that of the anti-apoptotic Bcl-2 gene decreased. Ddx3x was found to be the direct target of miR-322. MiR-424 was then detected in the seminal plasma of infertile patients with high DFI(DNA Fragmentation Index); this miRNA was down-regulated but Ddx3x was upregulated in the infertile group. Conclusion MiR-322 plays a key role in promoting GC-2 cell apoptosis by directly regulating Ddx3x expression. MiR-424 downregulation in infertile men may induce spermatogenic cell apoptosis and sperm DNA damage by directly acting on the target gene locus Ddx3x, resulting in male infertility.

Maternal effects of heat shock are reported for some species of insects, but little is known about such effects in the western flower thrips (WFT) Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). WFT is a pest of vegetables in greenhouses worldwide. It is susceptible to high temperatures in its natural environment and is controlled using heat treatment in China. WFT population growth is suppressed by a brief exposure to a high temperature of 40°C or 45°C in the laboratory. To explore the mechanism by which high temperatures suppress the growth of WFT populations, as well as the effects of multiple heat treatments on WFT, we recorded the duration of development and survival of immature WFT, and the sex ratio (female/male) and fecundity of F1, F2, F3 and F4 adult females that developed after a single heat shock, and those of F2 offspring after a double heat shock. We also recorded the longevity and ovarian structure of adult females of the treated generation (P) and their F1, F2 and F3 offspring after a single heat shock. In addition, we determined whether the effects of a heat shock on second instar nymphs and adults differed. The results indicate that exposure of the parental generation to 41°C or 45°C for 2 h significantly prolonged the duration of development, reduced survival of immature WFT and altered the sex ratio (female/male), longevity and fertility of their adult female offspring. The effects of a heat shock of 41°C persisted for two generations, whilst the effect of heat shock of 45°C persisted for three generations. In addition, double heat shocks had more pronounced effects than a single heat shock. Heat shock administered to second instar nymphs resulted in a decrease in the number of ovarioles, whilst a heat shock administered to adults resulted in ovariole deformity. The maternal effects of heat shock in terms of the biological parameters of WFT, structure and number of ovarioles, are critical in determining the suppression of the growth at high temperatures of WFT populations.

Silicon carbide (SiC) is a promising semiconductor material for making high-performance power electronics with higher withstand voltage and lower loss. The development of cost-effective machining technology for fabricating SiC wafers requires a complete understanding of the deformation and removal mechanism. In this study, molecular dynamics (MD) simulations were carried out to investigate the origins of the differences in elastic–plastic deformation characteristics of the SiC polytypes, including 3C-SiC, 4H-SiC and 6H-SiC, during nanoindentation. The atomic structures, pair correlation function and dislocation distribution during nanoindentation were extracted and analyzed. The main factors that cause elastic–plastic deformation have been revealed. The simulation results show that the deformation mechanisms of SiC polytypes are all dominated by amorphous phase transformation and dislocation behaviors. Most of the amorphous atoms recovered after completed unload. Dislocation analysis shows that the dislocations of 3C-SiC are mainly perfect dislocations during loading, while the perfect dislocations in 4H-SiC and 6H-SiC are relatively few. In addition, 4H-SiC also formed two types of stacking faults.

Background The age‐related decline in testosterone levels is thought to be of great importance for male aging and cardiovascular diseases. However, data are controversial on whether abnormal sex hormones are linked to the presence of cardiovascular diseases and it is also uncertain how blood pressure modifies the association between testosterone levels and major cardiovascular diseases. Methods and Results This is a multicenter, population‐based, cross‐sectional study of 6296 men conducted between 2013 and 2016. Basic information and clinical symptoms were obtained by questionnaires. Blood pressure and plasma levels of total testosterone, sex hormone–binding globulin, luteinizing hormone, and free testosterone were determined in men in a multistage random, cluster sampling in 6 provinces of China. There were 5786 Chinese men (mean [SD] age 55.0 [10.1] years) included after exclusion criteria were applied; 37.2% (2150) of them were diagnosed with hypertension. Total testosterone, free testosterone, and sex hormone–binding globulin were inversely associated with the prevalence of hypertension. Age >65 years or body mass index ≥24 negatively impacted the inverse correlation between testosterone levels and hypertension, whereas smoking and family history of hypertension strengthened the correlation. In participants with grade 2 hypertension, total testosterone was positively associated with the presence of stroke, and luteinizing hormone was also positively correlated with cardiovascular and cerebrovascular diseases. Conclusions Lower total testosterone could be a promising risk marker for prevalent hypertension. Both low and high levels of testosterone are associated with greater cardiovascular risk. Primary hypogonadism may be a risk marker for major cardiovascular diseases in men with severe hypertension.

Background: The current study aimed to determine the impact of SARS-CoV-2 infection on male fertility. Methods: This is a single-center, hospital-based observational study that included autopsied testicular and epididymal specimens of deceased COVID-19 male patients (n=6) and recruited recovering COVID-19 inpatients (n=23) with an equal number of age-matched controls, respectively. We performed histopathological examinations on testicular and epididymal specimens, and also performed TUNEL assay and immunohistochemistry. Whereas, we investigated the semen specimen for sperm parameters and immune factors. Findings: Autopsied testicular and epididymal specimens of COVID-19 showed the presence of interstitial edema, congestion, red blood cell exudation in testes, and epididymides. Thinning of seminiferous tubules was observed. The number of apoptotic cells within seminiferous tubules was significantly higher in COVID-19 compared to control cases. It also showed an increased concentration of CD3+ and CD68+ in the interstitial cells of testicular tissue and the presence of IgG within seminiferous tubules. Semen from COVID-19 inpatients showed that 39.1% (n=9) of them have oligozoospermia, and 60.9% (n=14) showed a significant increase in leucocytes in semen. Decreased sperm concentration, and increased seminal levels of IL-6, TNF-α, and MCP-1 compared to control males were observed. Interpretation: Impairment of spermatogenesis was observed in COVID-19 patients, which could be partially explained as a result of an elevated immune response in testis. Additionally, autoimmune orchitis occurred in some COVID-19 patients. Further research on the reversibility of impairment and developing treatment are warranted. Funding: This study was supported by Ministry of Science and Technology of China Plan, Hubei Science and Technology Plan, National Key Research and Development Program of China, HUST COVID-19 Rapid Response Call, China and National Natural Science Foundation of China; these funding bodies are public institutions, and they had no role in study conception, design, interpretation of results, and manuscript preparation.

At present, the etiology of most non-obstructive azoospermia (NOA) remains unclear. In vitro generation of patient-specific induced pluripotent stem cells (iPSCs) is an effective approach for exploring the mechanisms of human disease. Here, we established iPSCs from two patients with idiopathic NOA and differentiated them into primordial germ cell–like cells (PGCLCs) in vitro. Compared with iPSCs derived from normal fertile men, the NOA patient-specific iPSCs show decreased efficiency of PGCLC formation in vitro. Particularly, the embryoids derived from NOA patient-specific iPSCs show defects in the expression of early primordial germ cell (PGC) genes. The transcriptome analysis reveals the expression patterns of key human PGC genes are generally similar in PGCLCs differentiated from all iPSC lines, and the differentially expressed genes were enriched with gene ontology (GO) of cell cycle and apoptosis regulation. Moreover, the PGCLCs derived from NOA patient-specific iPSCs might have initiated epigenetic reprogramming at a very early stage. Thus, the NOA patient-specific iPSCs exhibit poor response to germ cell induction in vitro, which may be related to the regulation of apoptotic process. These findings provide a foundation for future research on mechanism of male infertility.

Sertoli cell-only syndrome (SCOS) is a severe phenotype of male infertility; autosomal gene defects are thought to be the causes for this disease. The iPSC line generated from a SCOS patient carrying a mutation in PIWIL2 gene expresses pluripotent markers, has a normal karyotype and the mutation c.731_732delAT in PIWIL2 gene and is able to differentiate into three germ layers. This cell line will help to study the pathogenesis of SCOS, and the roles of PIWIL2 in human germ cells development and spermatogenesis.

Abstract Background Serum leptin levels are augmented in obese infertile men and in men with azoospermia. They also correlate inversely with sperm concentration, motility and normal forms. The mechanisms underlying the adverse effects of excess leptin on male reproductive function remain unclear. The present study aimed to evaluate the effects of exogenous leptin on sperm parameters in mice and to explore the underlying mechanisms. Methods We treated normal adult male mice with saline, 0.1, 0.5 or 3 mg/kg leptin daily for 2 weeks. After treatment, serum leptin levels, serum testosterone levels, sperm parameters and testicular cell apoptosis were evaluated. Blood testis barrier integrity and the expression of tight junction-associated proteins in testes were also assessed. We further verified the direct effects of leptin on tight junction-associated proteins in Sertoli cells and the possible leptin signaling pathways involved in this process. Results After treatment, there were no significant differences in body weights, reproductive organ weights, serum leptin levels and serum testosterone levels between leptin-treated mice and control mice. Administration of 3 mg/kg leptin reduced sperm concentration, motility and progressive motility while increasing the percentage of abnormal sperm and testicular cell apoptosis. Mice treated with 3 mg/kg leptin also had impaired blood testis barrier integrity, which was related to decreased tight junction-associated proteins in testes. Leptin directly reduced tight junction-associated proteins in Sertoli cells, JAK2/STAT, PI3K and ERK pathways were suggested to be involved in this process. Conclusions Exogenous leptin negatively affects sperm parameters and impairs blood testis barrier integrity in mice. Leptin reduced tight junction-associated proteins in Sertoli cells, indicating that leptin has a direct role in impairing blood testis barrier integrity. Given the function of blood testis barrier in maintaining normal spermatogenesis, leptin-induced blood testis barrier impairment may be one of the mechanisms contributing to male subfertility and infertility.

The fifth-generation (5G) technology makes it widely applicable to connected vehicles. This would entail numerous transmitted data in communication networks and frequent information interactions between vehicles and other terminals, thus leading connected vehicles to be vulnerable to attacks from external communication interfaces. This paper analyzes potential security threats of 5G vehicular network and focuses on intrusion detection methods for in-vehicle networks. We choose four experiment scenarios from potential attacks for in-vehicle networks and collect real car data to compile various attack databases for the first time. In order to find appropriate methods to identify different attacks, four light-weight intrusion detection methods are presented to recognize abnormal behaviors of in-vehicle networks. Furthermore, our study undertakes the detection performance comparison between four detection methods with considering comprehensive evaluation metrics. The evaluation results provide optimal light-weight detection solution for in-vehicle networks. This paper facilitates the understanding of the advantages of the test methods in detection performance for in-vehicle networks and promotes the application of detection technology to deal with the security issues of automotive industry.

To clarify the influence mechanism of strain rate effect on deformation characteristics of aluminum nitride (AlN) ceramics, some varied-velocity nanoscratching tests were carried out using a Berkovich indenter in this paper. The deformation characteristics of the scratched grooves were observed using the scanning electron microscope. The experimental results showed higher scratch speed would lead to shallower penetration depth, fewer cracks, and indenter fewer slipping, which was more conducive to the plastic deformation of AlN ceramics. Considering the strain rate effect and the elastic recovery of material, a model for predicting the Berkovich indenter penetration depth under edge-forward mode was established. The prediction results were consistent with the experimental data, and the error was less than 5%, indicating that the model is effective. Based on the Boussinesq field, the Cerruti field, and the Sliding bubble field, a strain rate dependent scratch stress field model was established. The stress field revealed higher scratch speed may significantly reduce the maximum principal stress in the stress field under the indenter, which is the fundamental reason for reducing the crack damage and promoting the plastic deformation. The above study can provide theoretical guidance for reducing the processing damage of AlN ceramics.

A recent study found that three recurrent deletions of X chromosome linked copy number variations (CNVs), CNV64, CNV67 and CNV69 were associated with idiopathic male infertility in Spanish and Italian populations, especially CNV67 resembling the azoospermia factor deletions. That merits further investigations among different populations. This study was conducted to examine the prevalence of the three CNVs deletions and their associations with idiopathic male infertility in Chinese Han population. The present study included a large population of 1550 Chinese Han subjects recruited between 2014 and 2016. In total, 714 infertile participants were diagnosed as idiopathic infertility with different conditions (288 with non-obstructive azoospermia, 210 oligozoospermia and 216 asthenospermia) and 836 fertile participants (vasectomized men). The fertile participants were recruited from the representative areas: the north (Hebei and Shanxi), center (Hubei and Jiangsu), and south (Guangdong) of China. All patients were recruited from Hubei province. A multiplex PCR system was established to screen the deletion of the three CNVs, and deletion was confirmed by general PCR. Similar rates of these deletions were observed in infertile men and fertile participants (Hubei), and among the different conditions of infertility. Moreover, CNV64 and CNV67 map distribution geographically differed across China. The three CNVs in fertile groups of other regions were similar, except for Guangdong. No association between the three CNVs deletions and idiopathic male infertility was observed. CNV67 is rare in central China, albeit large sample size study for confirmation is warranted. It seems that the association between these CNVs deletions and idiopathic male infertility is ethnic dependent. There is still need to screen the CNVs deletions in other ethnicities. We suggested to consider the stratification patterns and geographic differences when prescribing CNVs deletions screening as a test in male infertility.

Aiming at the characteristics of the periodic stacking structure of a lithium-ion battery core and the corresponding relationship between the air-coupled ultrasonic transmission initial wave and the wave propagation mode in each layer medium of a lithium-ion battery, the homogenized finite element model of a lithium-ion battery was developed based on the theory of pressure acoustics and solid mechanics. This model provided a reliable method and basis for solving the visualization of ultrasonic propagation in a lithium-ion battery and the analysis of ultrasonic time-frequency domain characteristics. The finite element simulation analysis and experimental verification of a lithium-ion battery with a near-surface stomata defect, near-bottom stomata defect and middle-layer stomata defect were performed. The results showed that the air-coupled ultrasonic transmission signal can effectively characterize the stomata defect inside a lithium-ion battery. The energy of an air-coupled ultrasonic transmission signal is concentrated between 350−450 kHz, and the acoustic diffraction effect has an important influence on the effect of the ultrasonic and stomata defect. Based on the amplitude response characteristics of the air-coupled ultrasonic transmission wave in the stomata defect area, a C-scan of the lithium-ion battery was performed. The C-scan result verified that air-coupled ultrasonic testing technology can accurately and effectively detect the pre-embedded stomata defect and natural stomata defect in a lithium-ion battery, which is able to promote and expand the application of the technology in the field of electric energy security.

As a traditional Chinese drink, tea is favored for its rich flavor and its medicinal functionality. In this study, the in vitro bioactivities of Wushanshencha (WST; a local tea from Chongqing, China), which is processed mainly from the leaves of the wild Malus hupehensis (Pamp.) Rehd.). We assessed the scavenging capacity of tea extracts on 1, 1-diphenyl-2-picrylhydrazyl (DPPH); 2, 2′-azino-bis (3-ethylbenzthiazoline-6- sulphonic acid) diammonium salt (ABTS); and hydroxyl (OH) free radicals, and demonstrate the high antioxidant activity and dose-dependent relationship of these extracts. We also detail the anti-mutagenic effect of these tea extracts against the Salmonella typhimurium TA98 strain induced by the 2, 7-diaminofluorene (2, 7-AF) mutagen and the TA100 strain induced by the N-methyl-N′-nitro- N- nitrosoguanidine (MNNG) mutagen at concentrations of 1.25 and 2.50 mg/plate, respectively, with the high-dose groups showing better results. We investigated the anticancer mechanisms of WST extracts (40, 100, and 160 μg/mL) in HepG2 human hepatoma cells via 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay and quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR). The results showed that the proliferation of HepG2 cells was significantly inhibited in a dose-dependent manner by the tea extracts. Moreover, apoptosis in HepG2 cells was induced via upregulation of Caspase-3, Caspase-7, Caspase-8, Caspase-9, p21, p53, and Bax as well as downregulation of Bcl-2 apoptosis-associated factors, as assessed via mRNA expression levels after treating with WST extracts. The expression of inflammation-related factors, e.g., NF-κB, and Cox-2, was significantly downregulated by the WST extracts, demonstrating its inflammatory properties. Together, these observations indicated that WST extracts have anti-inflammatory and anti-cancer properties. In addition, high-performance liquid chromatography (HPLC) analysis showed that WST extracts contained chlorogenic acid, 4-hydroxycinnamic acid, isoquercitrin, taxifolin, quercitrin, rosmarinic acid, myricetin, baicalin, neosperidin dihydrochalcone, and quercetin. As such, WST appears to be an effectively functional drink, due to its rich functional components and anti-cancer activity.

The finite models of honeycomb sandwich composite with intact and embedded debonding defects are constructed. The sound pressure in fluid domain and the stress strain problem in solid domain are related by acoustic-structure coupling method, which visually shows the propagation process and modal characteristics of the acoustic wave inside the honeycomb sandwich composite. The simulation results show that the transmission longitudinal wave T1 (transmission initial wave) can effectively characterize debonding defects of honeycomb sandwich composite. However, in the actual detection of honeycomb sandwich composite, there are some problems, such as poor Signal-to-noise ratio (SNR) of received signal, incognizable transmission initial wave. In order to solve these problems, this paper proposes to apply polyphase coded pulse compression technique to air-coupled ultrasonic testing system. The actual test results show that the SNR of received signal is effectively improved, the transmission initial wave can be effectively identified, and the compressed signal has a good response to debonding defect. The air-coupled ultrasonic testing C scan result of honeycomb sandwich composite verifies the rationality and correctness of the theoretical simulation and signal processing technique, which promotes industrial application of air-coupled ultrasonic testing technique in the aerospace field.

[This corrects the article DOI: 10.1371/journal.pone.0164116.].

Decreased total testosterone (TT) is the recommended metric to identify age-related hypogonadism. However, average TT and the extent to which it varies by age, can vary substantially among different populations. Population-specific reference ranges are needed to understand normal versus abnormal TT levels. Therefore, the goal for this study was to describe androgen concentrations and their correlates among Western Chinese men. We completed a population-based, cross-sectional study including 227 young adults (YA) (20-39 years) and 939 older adults (OA) (40-89 years). We measured TT, sex-hormone binding globulin (SHBG), luteinizing hormone (LH), testosterone secreting index (TSI), and calculated free testosterone (cFT). Reference ranges for this population were determined using average YA concentrations. Multivariable regression models were used to predict hormone concentrations adjusting for age, waist-to-height ratio (WHR), marital status, education, occupation, smoking, alcohol, blood glucose, and blood pressure. Among OA, 3.8% had low TT, 15.2% had low cFT, 26.3% had low TSI, 21.6% had high SHBG, and 6.1% had high LH. Average cFT was significantly lower in OA (0.30 nmol/L; standard deviation (SD): 0.09) versus YA (0.37; SD: 0.11) but TT was not different in OA (16.82 nmol/L; SD: 4.80) versus YA (16.88; SD: 5.29). In adjusted models increasing age was significantly associated with increased SHBG or LH, and decreased cFT or TSI; however, TT was not significantly associated with age (β = 0.02 nmol/L; 95% confidence interval (CI): -0.01, 0.04). Higher WHR was associated with significantly decreased TT, SHBG, TSI, and LH. The only variable significantly related to cFT was age (β = -0.0033; 95% CI:-0.0037, -0.0028); suggesting that cFT measurements would not be confounded by other lifestyle factors. In conclusion, cFT, but not TT, varies with age in this population, suggesting cFT may be a better potential marker for age-related androgen deficiency than TT among Western Chinese men.

The mouse is an established and popular animal model for studying reproductive biology. Epididymal mouse sperm, which lack exposure to secretions of male accessory glands and do not precisely represent ejaculated sperm for the study of sperm functions, have been almost exclusively used in studies. We compared ejaculated and epididymal sperm in an in vitro fertilization setting to examine whether ejaculated sperm enter cumulus-oocyte complexes more efficiently. In order to prepare sperm for fertilization, they were incubated under capacitating conditions. At the outset of incubation, ejaculated sperm stuck to the glass surfaces of slides and the incidences of sticking decreased with time; whereas, very few epididymal sperm stuck to glass at any time point, indicating differences in surface charge. At the end of the capacitating incubation, when sperm were added to cumulus-oocyte complexes, the form of flagellar movement differed dramatically; specifically, ejaculated sperm predominantly exhibited increased bending on one side of the flagellum (a process termed pro-hook hyperactivation), while epididymal sperm equally exhibited increased bending on one or the other side of the flagellum (pro-hook or anti-hook hyperactivation). This indicates that accessory sex gland secretions might have modified Ca2+ signaling activities in sperm, because the two forms of hyperactivation are reported to be triggered by different Ca2+ signaling patterns. Lastly, over time, more ejaculated than epididymal sperm entered the cumulus oocyte complexes. We concluded that modification of sperm by male accessory gland secretions affects the behavior of ejaculated sperm, possibly providing them with an advantage over epididymal sperm for reaching the eggs in vivo.

Secretory azoospermia is a severe form of male infertility caused by unknown factors. DAX-1 is predominantly expressed in mammalian reproductive tissues and plays an important role in spermatogenesis because Dax-1 knockout male mice show spermatogenesis defects. To examine whether DAX-1 is involved in the pathogenesis of secretory azoospermia in humans, we sequenced all of the exons of DAX-1 in 776 patients diagnosed with secretory azoospermia and 709 proven fertile men. A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified. Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1. This novel mutation (p. V385L) of DAX-1 is the first to be identified in association with secretory azoospermia, thereby highlighting the important role of DAX-1 in spermatogenesis.

This paper examines the emergence of complex volatility in dynamic asset markets when there are heterogeneous agents. A discrete formulation is studied with two categories of market participants, fundamentalist traders who buy when the asset price is below the fundamental value and sell when it is above and noise traders who use moving average technical trading rules that can lead them to chase trends. Agents switch from one type of strategy to the other according to relative returns. A variety of outcomes are studied using numerical simulation, including variation of market price responsiveness to changes in excess demand, in switching behavior, and the introduction of noise. Bifurcation analysis of certain parameters is presented.

BACKGROUND: The great interest in cell-free mRNA, microRNA (miRNA) as molecular biomarkers for clinical applications, and as 'signaling' molecules for intercellular communication highlights the need to reveal their physical nature. Here this issue was explored in human cell-free seminal mRNA (cfs-mRNA) and miRNA (cfs-miRNA). METHODOLOGY/PRINCIPAL FINDINGS: Selected male reproductive organ-specific mRNAs, miRNAs, and piRNAs were quantified by quantitative real-time PCR in all experiments. While the stability of cfs-miRNA assessed by time-course analysis (up to 24 h at room temperature) was similar with cfs-mRNA, the reductive changes between cfs-miRNA and cfs-mRNA after filtration and Triton X-100 treatment on seminal plasma were very different, implying their different physical nature. Seminal microvesicles (SMVs) were then recovered and proportions of cfs-mRNA and cfs-miRNA within SMVs were quantified. The amounts of SMVs- sequestered cfs-mRNAs almost were the same as total cfs-mRNA, and were highly variable depending on the different sizes of SMVs. But most of cfs-miRNA was independent of SMVs and existed in the supernatant. The possible form of cfs-miRNA in the supernatant was further explored by filtration and protease K digestion. It passed through the 0.10-µm pore, but was degraded dramatically after intense protease K digestion. CONCLUSIONS/SIGNIFICANCE: The predominant cfs-mRNA is contained in SMVs, while most cfs-miRNA is bound with protein complexes. Our data explained the stability of extracellular RNAs in human semen, and shed light on their origins and potential functions in male reproduction, and strategy of developing them as biomarkers of male reproductive system.

No-tillage planters need to be equipped with row cleaners to remove post-harvest plant residue from the seedbed. The two-disc row cleaners cannot effectively remove the plant residue at high speed because the working depth is unstable, which leads to poor seeding quality of the seeder. A straw cleaning unit with double air springs was designed to achieve better straw cleaning performance at high speeds. The analysis of the mechanical characteristics of the double air spring system showed that it enabled separate adjustment of force and stiffness. A dynamic model of the straw cleaning unit was established, and the effectiveness of the double air spring system with adjustable stiffness in stabilizing the working depth of the row cleaners was analyzed. Field experiments were conducted to evaluate the straw cleaning performance and consistency of downforce against the ground of the straw cleaning unit with double air springs at different high speeds. The results showed that the stiffness of the double air spring system for better straw cleaning performance of the straw cleaning unit was different at different working speed, and the required stiffness increased as the working speed increased; When the working speed was 8- 12 km/h, the coefficient of variation of cleaned strip width was 6.9%-12.1%, the straw cleaning rate was 81.6%-92.2% and the root mean square error of downforce was 19.93-28.63 N; the coefficient of variation of cleaned strip width was moderately positively correlated with the root mean square error of downforce, and the cleaned strip width consistency was better when the root mean square error of downforce was lower than 25.00 N. [ABSTRACT FROM AUTHOR]