Caspase-1 Regulates the Apoptosis and Pyroptosis Induced by Phthalocyanine Zinc-Mediated Photodynamic Therapy in Breast Cancer MCF-7 Cells.

Photodynamic therapy (PDT) is an innovative and perspective antineoplastic therapy. Tetra-α-(4-carboxyphenoxy) phthalocyanine zinc (TαPcZn)-mediated PDT (TαPcZn-PDT) has shown antitumor activity in some tumor cells, but the manner in which caspase-1 is involved in the regulation of apoptosis and pyroptosis in the TαPcZn-PDT-treated breast cancer MCF-7 cells is unclear. Therefore, effects of TαPcZn-PDT on cytotoxicity, cell viability, apoptosis, pyroptosis, cellular reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), caspase-1, caspase-3, and nuclear transcription factor-κB (NFκB) in MCF-7 cells was firstly examined in the present study. The findings demonstrated that TαPcZn-PDT resulted in the increase in cytotoxicity and the percentage of apoptotic and pyroptotic cells, the reduction in cell viability and ΔΨm, the production of ROS and the activation of caspase-1, caspase-3 and NFκB in MCF-7 cells. Furthermore, the results also revealed that siRNA-targeting caspase-1 (siRNA-caspase-1) attenuated the effect of TαPcZn-PDT on apoptosis, pyroptosis and the activation of caspase-1, caspase-3 and NFκB in MCF-7 cells. Taken together, we conclude that caspase-1 regulates the apoptosis and pyroptosis induced by TαPcZn-PDT in MCF-7 cells.

Homozygous mutation in SLO3 leads to severe asthenoteratozoospermia due to acrosome hypoplasia and mitochondrial sheath malformations.

BACKGROUND: Potassium channels are important for the structure and function of the spermatozoa. As a potassium transporter, the mSlo3 is essential for male fertility as Slo3 knockout male mice were infertile with the series of functional defects in sperm cells. However, no pathogenic variant has been detected in human SLO3 to date. Here we reported a human case with homozygous SLO3 mutation. The function of SLO3 in human sperm and the corresponding assisted reproductive strategy are also investigated. METHODS: We performed whole-exome sequencing analysis from a large cohort of 105 patients with asthenoteratozoospermia. The effects of the variant were investigated by quantitative RT-PCR, western blotting, and immunofluorescence assays using the patient spermatozoa. Sperm morphological and ultrastructural studies were conducted using haematoxylin and eosin staining, scanning and transmission electron microscopy. RESULTS: We identified a homozygous missense variant (c.1237A > T: p.Ile413Phe) in the sperm-specific SLO3 in one Chinese patient with male infertility. This SLO3 variant was rare in human control populations and predicted to be deleterious by multiple bioinformatic tools. Sperm from the individual harbouring the homozygous SLO3 variant exhibited severe morphological abnormalities, such as acrosome hypoplasia, disruption of the mitochondrial sheath, coiled tails, and motility defects. The levels of SLO3 mRNA and protein in spermatozoa from the affected individual were reduced. Furthermore, the acrosome reaction, mitochondrial membrane potential, and membrane potential during capacitation were also afflicted. The levels of acrosome marker glycoproteins and PLCζ1 as well as the mitochondrial sheath protein HSP60 and SLO3 auxiliary subunit LRRC52, were significantly reduced in the spermatozoa from the affected individual. The affected man was sterile due to acrosome and mitochondrial dysfunction; however, intra-cytoplasmic sperm injection successfully rescued this infertile condition. CONCLUSIONS: SLO3 deficiency seriously impact acrosome formation, mitochondrial sheath assembly, and the function of K+ channels. Our findings provided clinical implications for the genetic and reproductive counselling of affected families.

Identification and validation of glomerulotubular crosstalk genes mediating IgA nephropathy by integrated bioinformatics.

BACKGROUND: IgA nephropathy (IgAN), which has been reported as the most prevalent glomerulonephritis globally, is the major contributor to end-stage renal diseases. This bioinformatics study aimed to explore glomerulotubular crosstalk genes and dysregulated pathways relating to the pathogenesis of IgAN. METHODS: The microarray datasets from the Gene Expression Omnibus (GEO) database were searched. Weighted gene co-expression network analysis (WGCNA) and differentially expressed genes (DEGs) of both glomeruli and tubulointerstitium were conducted individually. The co-expression gene modules of glomeruli and tubulointerstitium were compared via gene function enrichment analysis. Subsequently, the crosstalk co-expression network was constructed via the STRING database and key genes were mined from the crosstalk network. Finally, key genes were validated using another GEO dataset (GSE99340) containing RNA-seq data of IgAN and lupus nephritis, and their potential diagnostic values were shown using receiver operating characteristic (ROC) analysis. RESULTS: Five hundred eighty-three DEGs and eight modules were identified in glomerular samples, while 272 DEGs and four modules were in tubulointerstitial samples. There were 119 overlapping DEGs between the two groups. Among the distinctive modules, four modules in glomeruli and one module in tubulointerstitium were positively associated with IgAN. While four modules in glomeruli and two modules in tubulointerstitium were negatively associated with IgAN. The top ten key genes screened by CytoHubba were ITGAM, ALB, TYROBP, ITGB2, CYBB, HCK, CSF1R, LAPTM5, FN1, and CTSS. Compared with lupus nephritis, there were significant differences in the expression levels of CYBB, CTSS and TYROBP (P < 0.05), while other key genes showed no significant difference. Meanwhile, CYBB, CTSS, and TYROBP demonstrated possible diagnostic significance. CONCLUSIONS: The crosstalk genes confirmed in this study may provide novel insight into the pathogenesis of IgAN. Immune-related pathways are associated with both glomerular and tubulointerstitial injuries in IgAN. The glomerulotubular crosstalk might perform a role in the pathogenesis of IgAN.

EPO activates PI3K-IKKα-CDK1 signaling pathway to promote the proliferation of Glial Cells under hypoxia environment.

Erythropoietin (EPO), supports the function and survival of neurons through astrocytes and has a protective role in neonatal asphyxia brain injury; yet, its mechanism of action remains unclear. As a neuroprotective factor, EPO is also used in the treatment of various diseases, such as neurodegenerative diseases, Parkinson's disease, traumatic brain injury, by decreasing inflammatory reaction, resisting apoptosis, and lowering oxidative stress. The aim of this study was to examine the effect and mechanism of EPO on promoting human brain glial cell proliferation under hypoxia in vitro. Under CoC12-induced hypoxia, after adding EPO, high-throughput sequencing was used to screen out meaningful up-regulated and significant differentially expressed genes PI3K, IKKα CDK1 related to proliferation, and make further verification by qPCR and western blotting. Under hypoxia, EPO promoted cell proliferation and the expression of PI3K while this effect was inhibited (along with a decrease of downstream genes IKKα and CDK1 decreased) after adding PI3K inhibitor to cell culture. EPO can promote cell proliferation and CDK1 expression, while after inhibiting CDK1 expression, the promotion of EPO on cell proliferation was eliminated. These data proved that EPO promotes the proliferation of U251 cells by activating the PI3K-IKKα-CDK1 signaling pathway under CoC12-induced hypoxia.

[Causes of failure in autologous sperm preservation in the human sperm bank and countermeasures].

OBJECTIVE: To analyze the causes of failure in autologous sperm cryopreservation (ASCP) in the human sperm bank and propose some countermeasures to improve the success rate of ASCP and promote it in human sperm banks. METHODS: We retrospectively analyzed the reasons for and causes of failure in ASCP 1 156 males undergoing ASCP in the Human Sperm Bank of Guangdong Province from January 1, 2016 to December 31, 2019. RESULTS: Of the 1 156 cases of ASCP, 857 (74.1%) succeeded and 299 (25.9%) failed, with a failure rate of 56.1% (96/171) in the microdissection testicular sperm extraction (micro-TESE) group, 29.9% (67/224) in the reproduction insurance group, 21.2% (27/100) in the non-tumor disease group and 17.2% (109/525) in the tumor group, with statistically significant difference between the four groups (χ2 = 109.926, P < 0.01). The causes of failed ASCP included failure to extract semen (14.0% ï¼»42/299ï¼½), failure to meet the standard of sperm cryopreservation (67.6% ï¼»202/299ï¼½), giving up ASCP for low semen quality (7.4% ï¼»22/299ï¼½), and giving up ASCP for some other reasons (11.0% ï¼»33/299ï¼½), including worry about the reliability of cryopreservation technology (6.0% ï¼»18/299ï¼½), suspicion about the complexity of the ASCP procedures (3.0% ï¼»9/299ï¼½) and expectation for fertility recovery after chemotherapy withdrawal (2.0% ï¼»6/299ï¼½). CONCLUSIONS: In view of different causes of failure in ASCP, human sperm banks can provide individualized cryopreservation schemes, including guidance with masturbation or the use of sperm extraction instrument in sperm extraction, strengthening sperm preservation-related education and the awareness of reproduction protection and earlier sperm preservation among cancer patients, promoting the cryopreservation of microsamples of motile sperm by microsample or single sperm freezing, and development of testis tissue cryopreservation to preserve the fertility of children with cancer.

A Pilot Study on the Effects of Yi Jin Jing Exercise for Older Men with Benign Prostatic Hyperplasia.

Benign prostatic hyperplasia and its associated lower urinary tract symptoms seriously affect both the physical and mental health of older men. In order to determine the efficiency of Chinese Qigong Yi Jin Jing on prostate health in older individuals, thirty participants were randomized into either an Yi Jin Jing group (n = 15) or a control group (n = 15). After the 6-month intervention, the Yi Jin Jing group showed a significant decrease in international prostate symptom score and a significant increase in maximal urinary flow rate (compared with the control group p = .005, p = .001, respectively). Also, testosterone level increased and estrogen/testosterone ratio decreased in the Yi Jin Jing group (compared with the baseline p = .004, p = .002, respectively); estrogen level and estrogen/testosterone ratio were lower in the Yi Jin Jing group (compared with the control group p = .029, p = .012, respectively). The results showed that Yi Jin Jing is a promising way to reduce the risk of benign prostatic hyperplasia-lower urinary tract symptoms in older men.

Circular RNA circ_001350 regulates glioma cell proliferation, apoptosis, and metastatic properties by acting as a miRNA sponge.

Glioma is an aggressive malignancy with increasing incidence and threatens people's health worldwide. Accumulating evidence revealed that circular RNAs (circRNAs) play important functions in cancers. A previous study demonstrated that circ_001350 was elevated in glioma tissue samples than nontumorous tissue specimens screened by high-throughput microarray. The level of circ_001350 in glioma tissue specimens and cell lines was detected by quantitative real-time polymerase chain reaction. The Fisher exact test was carried out to estimate the correlation of circ_001350 level with clinical characteristics. Cell proliferation, apoptosis, and motility abilities were detected using cell counting kit-8, clonogenic, flow cytometry, and transwell experiments, respectively. The potential target of circ_001350 was identified by the luciferase assay. circ_001350 level was significantly enhanced in glioma tissue specimens and cells. Further, elevated expression of circ_001350 was closely linked to patients' clinical severity. Knockdown of circ_001350 could inhibit cell proliferation and metastatic properties and increase apoptotic cells. circ_001350 could directly bind to miR-1236 and regulate its expression to exert oncogenic functions. Collectively, circ_001350 directly sponges miR-1236, thus contributing to malignant progression of glioma.

[Differentially expressed long non-coding RNAs in spermatozoa with different fertilizing abilities].

OBJECTIVE: To search for conception capacity-related long non-coding RNAs (lncRNA) and explore their possible roles in fertilization. METHODS: We obtained 10 semen samples, 5 of high and the other 5 of low fertilizing ability, extracted large RNAs, established a cDNA library, and performed RNA sequencing with the HiSeq 2000 sequencing system. Using the bioinformatics method, we assembled and predicted lncRNAs, screened differentially expressed genes between the two groups by NOIseq, analyzed the lncRNAs with the box plot and volcano plot, and determined their expression patterns by hierarchical cluster analysis. We examined the functional classification of differentially expressed lncRNAs by pathway and gene ontology (GO) enrichment and predicted those of some lncRNAs by lncRNA-mRNA interaction analysis and intersection analysis with up- and down-stream cis-acting elements. RESULTS: A total of 147 1615 lncRNAs were identified in all the semen samples, including 463 596 novel ones and 8 019 known ones, with 4 052 differentially expressed lncRNAs, 985 upregulated and the other 3 067 downregulated. Box plot and volcano plot filtering analyses showed statistically significant differences in the expressions of the lncRNAs between the two groups, and so did hierarchical cluster analysis. GO functional annotations manifested the involvement of the differentially expressed lncRNAs in the metabolic process, biological regulation, membrane and organelle formation, and protein-nucleotide binding. Pathway analysis showed that the differentially expressed lncRNAs were related to transport and catabolism, cell motility, signaling molecular interactions, signaling transduction, and signaling pathways in the development and immune systems. The functions of the 5 lncRNAs predicted were shown to be associated with sperm motility, acrosomal reaction and signal transduction during fertilization. CONCLUSIONS: Differentially expressed lncRNAs may play an important role in fertilization and become biomarkers for the assessment of sperm quality.

Isolation and antiviral activity of water-soluble Cynomorium songaricum Rupr. polysaccharides.

The plant, Cynomorium songaricum Rupr., is used as a traditional medicine in China and Mongolia. In the present study, two new water-soluble polysaccharides isolated from C. songaricum Rupr. were purified by successive Sephadex G-75 and G-50 column chromatographies and then characterized by high resolution NMR and IR spectroscopies. The molecular weights of two polysaccharides were determined by an aqueous GPC to be [Formula: see text] = 3.7 × 10(4) and 1.0 × 10(4), respectively. In addition, it was found that the polysaccharide with the larger molecular weight was an acidic polysaccharide. It was found that the iodine-starch reaction of both isolated polysaccharides was negative and the methylation analysis gave 2, 4, 6-tri-O-methyl alditol acetate as a main product. NMR and IR measurements and sugar analysis revealed that both polysaccharides had a (1 â†’ 3)-α-d-glucopyranosidic main chain with a small number of branches. After sulfation, the sulfated C. songaricum Rupr. polysaccharides were found to have a potent inhibitory effect on HIV infection of MT-4 cells at a 50% effective concentration of 0.3-0.4 µg/ml, a concentration that has almost the same high activity as standard dextran and curdlan sulfates, EC50 = 0.35 and 0.14 µg/ml, respectively. The 50% cytotoxic concentration was low, CC50>1000 µg/ml. In addition, the interaction between the sulfated polysaccharides and poly-l-lysine as a model protein compound was investigated by a surface plasmon resonance to reveal the anti-HIV mechanism.

Multi-layered carbon accommodation of MnO2 enabling fast kinetics for highly stable zinc ion batteries.

Large-scale durable aqueous zinc ion batteries for stationary storage are realized by spray-coating conductive PEDOT(Poly(3,4-ethylenedioxythiophene)) wrapping MnO2/carbon microspheres hybrid cathode in this work. The porous carbon microspheres with multiple layers deriving from sucrose provide suitable accommodation for MnO2 active materials, exposing more redox active sites and enhancing the contact surface between electrolyte and active materials. As a result, MnO2/microspheres are adhered to the current collector by a conductive PEDOT coating without any binder. The ternary design retards the structural degradation during cycling and shortens the electron and ion transport path, rendering the full batteries high capacity and long cycle stability. The resulting batteries perform the capacity of 277, 227, 110, 85 and 50 mAh/g at 0.2, 0.5, 1, 2 and 5 A/g, respectively. After 3000 cycles the initial capacity retains 86%, and 80% after 5000 cycles. GITT indicates PEDOT wrapping MnO2/microspheres cathode enables better ion intercalating kinetics than conventional MnO2. The work could represent a novel and significant step forward in the studies on the large-scale application of zinc ion batteries.

Copper-dependent control of uptake, translocation and accumulation of cadmium in hyperaccumlator Sedum alfredii.

Cadmium (Cd) is detrimental to plant growth and threatens human health. Here, we investigated the potential for remediation of Cd-contaminated soil with high copper (Cu) background using Cd hyperaccumulator ecotype (HE) Sedum alfredii. We assessed effects of Cu on Cd accumulation, compartmentation and translocation in HE S. alfredii, and compared with those in a related non-accumulator ecotype (NHE). We found that Cu supply significantly induced Cd accumulation in roots and shoots of long-term soil-cultivated HE S. alfredii. A large fraction of root Cd was accumulated in the organelles, but a small fraction was stored in the cell wall. Importantly, Cu addition reduced Cd accumulation in the cell wall and the organelles in root cells. Furthermore, leaf cell capacity to sequestrate Cd in the organelles was greatly improved upon Cu exposure. We also found that genes involving metal transport and cell wall remodeling were distinctly regulated to mediate Cd accumulation in HE S. alfredii. These findings indicate that Cu-dependent decrease of root cell-wall-bound Cd, and stimulation of efflux/influx of organelle Cd transport in root and leaf cells plays a role in the dramatic Cd hyperaccumulation expressed in naturally survived HE S. alfredii.

Traditional Use, Phytochemistry, Pharmacology, Toxicology and Clinical Applications of Persicae Semen: A Review.

Persicae Semen (Taoren), the seed of mature peaches consumed as both food and medicine, is native to the temperate regions of China, distributed in the provinces of North and East China, and currently cultivated worldwide. The primary components of Persicae Semen include volatile oil, protein, amino acids, amygdalin, and prunasin, all of which have pharmacological properties, such as anti-inflammatory, antioxidant, and immune regulatory effects, and are clinically used in the treatment of gynecological, cardiovascular, cerebrovascular, orthopedic, and digestive system diseases. This review provides a comprehensive perspective on the resource status, ethnopharmacology, phytochemistry, pharmacology, and toxicology, as well as the trend of Persicae Semen patent, global distribution, and clinical applications. This review will help facilitate the development and utilization of Persicae Semen in clinical settings.

Arabidopsis COPT1 copper transporter uses a single histidine to regulate transport activity and protein stability.

Copper acquisition and subsequent delivery to target proteins are essential for many biological processes. However, the cellular levels of this trace element must be controlled because of its potential toxicity. The COPT1 protein rich in potential metal-binding amino acids functions in high affinity copper uptake at the plasma membrane of Arabidopsis cells. The functional role of these putative metal-binding residues is largely unknown. Through truncations and site-directed mutagenesis, we identified His43, a single residue within the extracellular N-terminal domain as absolutely critical for copper uptake of COPT1. Substitution of this residue with leucine, methionine or cysteine almost inactivated transport function of COPT1, implying that His43 fails to serves as a copper ligand in the regulation of COPT1 activity. Deletion of all extracellular N-terminal metal-binding residues completely blocked copper-stimulated degradation but did not alter the subcellular distribution and multimerization of COPT1. Although mutation of His43 to alanine and serine retained the transporter activity in yeast cells, the mutant protein was unstable and degraded in the proteasome in Arabidopsis cells. Our results demonstrate a pivotal role for the extracellular residue His43 in high affinity copper transport activity, and suggest common molecular mechanisms for regulating both metal transport and protein stability of COPT1.

Chemotherapy-Sensitized In Situ Vaccination for Malignant Osteosarcoma Enabled by Bioinspired Calcium Phosphonate Nanoagents.

How to effectively treat malignant osteosarcoma remains clinically challenging. Programmed delivery of chemotherapeutic agents and immunostimulants may offer a universal strategy for killing osteosarcoma cells while simultaneously eliciting in situ antitumor immunity. However, targeted chemoimmunotherapy lacks a reliable delivery system. To address this issue, we herein developed a bioinspired calcium phosphonate nanoagent that was synthesized by chemical reactions between Ca2+ and phosphonate residue from zoledronic acid using bovine serum albumin as a scaffold. In addition, methotrexate combination with a phosphorothioate CpG immunomodulator was also loaded for pH-responsive delivery to enable synergistic chemoimmunotherapy of osteosarcoma. The calcium phosphonate nanoagents were found to effectively accumulate in osteosarcoma for nearly 1 week, which is favorable for exerting the vaccination effects in situ by maturing dendritic cells and priming CD8+ T cells to suppress the osteosarcoma progression and pulmonary metastasis through controlled release of the three loaded agents in the acidic tumor microenvironment. The current study may thus offer a reliable delivery platform for achieving targeted chemotherapy-induced in situ antitumor immunity.

Study on the Work Hardening and Metamorphic Layer Characteristics of Milling Contour Bevel Gears.

High temperature and strain will occur in the cutting area during dry milling of contour bevel gears, which causes plastic deformation of the workpiece, resulting in changes in the physical properties of the machined surface's metamorphic layer, reducing the quality of the workpiece's machined surface. Therefore, it is necessary to investigate the properties of the metamorphic layer and the work hardening behavior of the machined surfaces of contour bevel gears. The paper first establishes a single-tooth finite element simulation model for a contour bevel gear and extracts the temperature field, strain field and strain rate at different depths from the machined surface. Then, based on the simulation results, the experiment of milling contour bevel gears is carried out, the microscopic properties of the machined metamorphic layer are studied using XRD diffractometer and ultra-deep field microscopy, and the work hardening behavior of the machined metamorphic layer under different cutting parameters is studied. Finally, the influence of the cutting parameters on the thickness of the metamorphic layer of the machined surface is investigated by scanning electron microscopy. The research results can not only improve the surface quality and machinability of the workpiece, but are also significant for increasing the fatigue strength of the workpiece.

Gadolinium-Bisphosphonate Nanoparticle-Based Low-Dose Radioimmunotherapy for Osteosarcoma.

Osteosarcoma is a malignant osteogenic tumor with a high metastatic rate commonly occurring in adolescents. Although radiotherapy is applied to treat unresectable osteosarcoma with radiation resistance, a high dose of radiotherapy is required, which may weaken the immune microenvironment. Therefore, there is an urgent need to develop novel agents to maximize the radiotherapeutic effects by eliciting immune activation effects. In this study, we synthesized therapeutic gadolinium-based metal-bisphosphonate nanoparticles (NPs) for osteosarcoma treatment that can be combined with radiotherapy. The gadolinium ion (Gd) was chelated with zoledronic acid (Zol), a commonly used drug to prevent/treat osteoporosis or bone metastases from advanced cancers, and stabilized by ovalbumin (OVA) to produce OVA-GdZol NPs. OVA-GdZol NPs were internalized into K7M2 osteosarcoma cells, showing a high sensitization effect under X-ray irradiation. Cell pretreatment of OVA-GdZol NPs significantly enhanced the radiation therapeutic effect in vitro by reducing the cell colonies and increased the signal of γH2AX-positive cells. More importantly, OVA-GdZol NPs promoted the maturation of bone marrow-derived dendritic cells (BMDCs) and M1 polarization of macrophages. The inhibitory effect on K7M2 osteosarcoma of OVA-GdZol NPs and X-ray radiation was evident, indicated by a significantly reduced tumor volume, high survival rate, and decreased lung metastasis. Meanwhile, both innate and adaptive immune systems were activated to exert a strong antitumor effect. The above results highly suggest that OVA-GdZol NPs serve as both radiosensitizers and immune adjuvants, suitable for the sequential combination of vaccination and radiotherapy.

Homozygous mutation in DNALI1 leads to asthenoteratozoospermia by affecting the inner dynein arms.

Asthenozoospermia is the most common cause of male infertility. Dynein protein arms play a crucial role in the motility of sperm flagella and defects in these proteins generally impair the axoneme structure and affect sperm flagella function. In this study, we performed whole exome sequencing for a cohort of 126 infertile patients with asthenozoospermia and identified homozygous DNALI1 mutation in one patient from a consanguineous family. This identified homozygous mutation was verified by Sanger sequencing. In silico analysis showed that this homozygous mutation is very rare, highly pathogenic, and very conserved. Sperm routine analysis confirmed that the motility of the spermatozoa from the patient significantly decreased. Further sperm morphology analysis showed that the spermatozoa from the patient exhibited multiple flagella morphological defects and a specific loss in the inner dynein arms. Fortunately, the patient was able to have his child via intracytoplasmic sperm injection treatment. Our study is the first to demonstrate that homozygous DNALI1 mutation may impair the integration of axoneme structure, affect sperm motility and cause asthenoteratozoospermia in human beings.

Circulating microRNAs in seminal plasma as predictors of sperm retrieval in microdissection testicular sperm extraction.

Background: Because of focal spermatogenesis in some nonobstructive azoospermia (NOA) patients, testicular spermatozoa can be retrieved by microdissection testicular sperm extraction (micro-TESE) for intracytoplasmic sperm injection (ICSI) to achieve successful fertilization. Currently, testicular biopsy is widely performed for the prognosis of micro-TESE; however, it might miss foci with active spermatogenesis because of the 'blind manner' of puncture, highlighting the needs for biomarkers that could indicate actual spermatogenesis conditions in the testis. Thus, we screened microRNAs in the seminal plasma for potential biomarkers to provide a non-invasive and reliable preoperative assessment for micro-TESE. Methods: We screened the seminal plasma microRNAs from NOA patients with and without sperm retrieval (n=6 in each group) together with fertile men (n=6) by RNA sequencing, and the selected microRNAs were validated by quantitative polymerase chain reaction (qPCR). Next, a predictive model was established by performing ordered logistic regression using the qPCR data of 56 specimens, and the predictive accuracy of this model was evaluated using 40 more specimens in a blind manner. Results: Four microRNAs (hsa-miR-34b-3p, hsa-miR-34c-3p, hsa-miR-3065-3p, and hsa-miR-4446-3p) were identified as biomarkers, and the predictive model Logit = 2.0881+ 0.13448 mir-34b-3p + 0.58679 mir-34c-3p + 0.15636 mir-3065-3p + 0.09523 mir-4446-3p was established by machine learning. The model provided a high predictive accuracy (AUC =0.927). Conclusions: We developed a predictive model with high accuracy for micro-TESE, with which NOA patients might obtain accurate assessment of spermatogenesis conditions in testes before surgery.

Evaluating an external quality assurance program for semen analysis in China during 2009-2020.

INTRODUCTION: Semen analysis (SA) plays a key role in guiding treatments of male reproductive diseases and infertility due to male factors; however, it remains challenging to conduct an accurate SA due to lack of standardization, highly subjective assessments, and problems with automated procedures. Therefore, quality assurance (QA) and teaching courses are essential for making the laboratory results more consistent. MATERIALS AND METHODS: The external quality assurance (EQA) scheme was organized by national human sperm bank technology training bases in Guangdong province in China between 2009 and 2020. Until 2020, 124 laboratories from China participated in the EQA program. The EQA scheme per year has been organized involving two semen aliquots for sperm concentration, two video recordings for motility, and two smears for sperm morphology. All samples used in the EQA scheme were obtained from different healthy donors or patients. RESULTS: We estimated that the median coefficient of variation (CV) of sperm concentration, ignoring the method used, was 26.6%. Using a 100 µm deep counting chamber led to a decreasing CV of 13.6%. For sperm motility, the median CV of nonprogressive motility was high (50.8%), but the CV of progressive motility (13.2%), immotile sperm (14.3%), and total motility (11.8%) were acceptable. The morphology assessment revealed large variability (44.4%) irrespective of the classification criteria. DISCUSSION: The reduction of interlaboratory variability is still a challenge during SA in China. Therefore, it is critical to increase awareness of joining EQA schemes and establish standardized training centers to follow WHO-recommended procedures toward Chinese standards.

Effects of varicocele and microsurgical varicocelectomy on the metabolites in semen.

The influence of varicocele and microsurgical varicocelectomy on semen quality remains unclear. Few studies have investigated the relationship between semen metabolism and the abnormalities in reproductive function caused by varicocele, however, there is no study on the changes of semen metabolism after microsurgical varicocelectomy. Here, we used the non-targeted and targeted metabolic analysis to investigate the different metabolites in seminal plasma within normal, varicocele, and varicocelectomy groups. We clearly showed that varicocele significantly affects semen metabolism, and microsurgical varicocelectomy can reverse this metabolic abnormality. Moreover, we characterized the landscape of three dipeptides in the seminal plasma of patients with varicocele that have not been identified previously in human tissues or biofluids. Interestingly, the levels of these three dipeptides decreased after microsurgical varicocelectomy coincident with an improvement in semen quality. Western blotting confirmed the downregulation of DPEP3 (dipeptidase 3) in the varicocele group and the upregulation of DPEP3 in the varicocelectomy group. Furthermore, we found that eight metabolites may be helpful to distinguish varicocele patients from normal subjects. Our results may be applied to earlier diagnosis or to predict the outcome of microsurgery for varicocele.