Ureaplasma urealyticum infection and apoptosis of spermatogenic cells

Aim: To study the relationship between Ureaplasma urealyticum (UU) infection and apoptosis of human spermato-genic cells. Methods: Spermatogenic cells were observed under light microscope with Wright-Giemsa staining andby means of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling(TUNEL) technique. Results: Apoptotic rate of UU-infected males (15.5 % ± 6.8 % ) was significantly higherthan that of controls (5.2 % ± 2.3 % ). Conclusion: Apoptosis of spermatogenic cells can be caused by UU in-fection, which provides further evidence for UU-induced male infertility. (Asian J Androl 1999 Sep ; 1: 127 - 129)

Quantitative (stereological) study of incomplete spermatogenic suppression induced by testosterone undecanoate injection in rats

Aim: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. Methods: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector. Results: In response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged. Conclusion: Double inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gona-dotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels.

Gene functional research using polyethylenimine-mediated in vivo gene transfection into mouse spermatogenic cells

Aim： To study polyethylenimine （PEI）-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method. Methods： PEI/DNA complexes were introduced into the seminiferous tubules of mouse testes using intratesticular injection. Transfection efficiency and speciality were analyzed on the third day of transfection with fluorescent microscopy and hematoxylin staining. The long-lasting expression of the GFP-NYD-SP12 fusion protein and its subcelluar localization in spermatogenic cells at different stages were analyzed with fluorescent microscopy and propidium iodide staining. Results： With the mediation of PEI, the GFP-NYD-SP12 fusion gene was efficiently transferred and expressed in the germ cells （especially in primary spermatocytes）. Transfection into Sertoli cells was not observed. The subcellular localization of the GFP-NYD-SP2 fusion protein showed dynamic shifts in spermatogenic cells at different stages during spermatogenesis. Conclusion： PEI can efficiently mediate gene transfer into spermatocytes. Thus, it might be useful for the functional research of spermatogenic-cell specific genes such as the NYD-SP12 gene. In our gtudy, the NYD-SP12 protein was visualized and was involved in the formation of acrosome during spermatogenesis. Our research will continue into the detailed function of NYD-SP12 in spermatocytes. （Asian J Androl 2006 Jan; 8： 53-59）

Simulated Microgravity Conditions and Carbon Ion Irradiation Induce Spermatogenic Cell Apoptosis and Sperm DNA Damage

Objective To investigate the effect of simulated microgravity and carbon ion irradiation （CIR） on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment. Methods Sperm DNA damage indicated by DNA fragmentation index （DFI） and high DNA stainability （HDS） was measured by sperm chromatin structure assay （SCSA）. Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax （the expression levels of p53） and proliferating cell nuclear antigen （PCNAI were measured by immunoblotting; p53 and PCNA were located by immunohistology. Results HDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups （P〈0.05） compared with those in the control group, however, the PCNA expression varied to a certain degree, p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes. Conclusion The findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protectint~ astronauts and space traveler＇s health and safety.

Association of USP26 haplotypes in men in Taiwan, China with severe spermatogenic defect

Aim： To complete comprehensive haplotype analysis of USP26 for both fertile and infertile men. Methods： Two hundred infertile men with severe oligospermia or non-obstructive azoospermia were subjected to sequence analysis for the entire coding sequences of the USP26 gene. Two hundred men with proven fertility were genotyped by primer extension methods. Allele/genotype frequencies, linkage disequilibrium （LD） characteristics and haplotypes of fertile men were compared with infertile men. Results： The allele frequencies of five single nucleotide polymor- phisms （370-37 linsACA, 494T〉C, 576G〉A, ss6202791C〉T, 1737G〉A） were significantly higher in infertile patients than control subjects. The major haplotypes in infertile men were TACCGA （28% of the population）, TGCCGA （15%）, TACCAA （8%）, TGCCAA （6%）, TATCAA （5%） and CATCAA （5%）. The major haplotypes for the control subjects were TACCGA （58% of the population）, CACCGA （7%）, CATCGA （6%） and TGCCGA （5%）. Haplotypes TGCCGA, TATCAA, CATCAA, CATCGC, TACCAA and TGCCAA were over-transmitted in patients with spermato- genic defect, whereas haplotypes TACCGA, CACCGA, and CATCGA were under-transmitted in these patients. Conclusion： Some USP26 alleles and haplotypes are associated with spermatogenic defect in the Han nationality in Taiwan, China.

LM23 is a novel member of the Speedy/Ringo family at the ：rossroads of life and death of spermatogenic cell

LM23 is a gene specifically expressed in the testis of Rattus norvegicus, as previously reported by our laboratory. The aim of the study is to further investigate the biological function of LM23. Several bioinformatic tools were utilized, including PROSITE and BLAST. To determine the subcellullar localization of LM23, a polyclonal antibody specific for LM23 was generated via the immunization of rabbits. The LM23gene was cloned from rat testis tissue, and LM23 protein was expressed in Escherichia co/i. The biological function of LM23 was analyzed with microarray analysis and immunohistochemistry, using a rat model of LM23 gene knockdown. The results suggested that LM23 belongs to the Speedy/Ringo family. LM23 regulated the GI/S and G2/M transitions of the cell cycle during spermatogenesis. Downregulation of the LM23gene during spermatogenesis could lead to the activation of both the Fas-FasL pathway and the mitochondrial pathway. These novel findings indicate that LM23 has a diverse array of functions that are important in both the life and death of the spermatogenic cell.

Medical therapy for spermatogenic failure

Medical treatment of men with primary spermatogenic failure remains largely ineffective in contrast to those with secondary testicular failure. Treatment has been attempted with a multitude of agents ranging from hormones to nutritional supplements （antioxidants）. While some studies have demonstrated benefit to some treatments, no treatments have consistently demonstrated efficacy nor has it been possible to reliably identify patients likely to benefit. Idiopathic spermatogenic failure likely results from multiple discrete defects in sperm production that are as yet unidentified. A better understanding of these defects will yield more effective treatment options and appropriate triage of patients to specific therapeutic regimens. This review focuses on the rationale and current evidence for hormonal and antioxidant therapy in medical treatment of male infertility, spermatogenic failure in particular. Although empiric medical therapy for spermatogenic failure has been largely replaced by assisted reproductive techniques, both treatment modalities could play a role, oerhaos as combination therapy.

20(R)-ginsenoside Rg3,a product of high-efficiency thermal deglycosylation of ginsenoside Rd,exerts protective effects against scrotal heat-induced spermatogenic damage in mice

Heat stress(HS)reaction can lead to serious physiological dysfunction associated with cardiovascular and various organ diseases.Ginsenoside Rg3(G-Rg3)is a representative component of ginseng rare saponin and can protect against multiple organs,also used as functional food to adjust the balance of the human body,but the therapeutic effect and molecular mechanism of G-Rg3 on male diseases under HS are underexplored.The aim of the present study,G-Rg3 was prepared through the efficient conversion of ginsenoside Rd and investigate the contribution of G-Rg3 to testicular injury induced exposure to HS.All mice were divided into four groups as follows:normal group,HS group,and HS+G-Rg3(5 and 10 mg/kg)groups.G-Rg3 was administered orally for 14 days,then exposed to a single scrotal heat treatment(43°C,18min)on the 7th day.After HS treatment,the morphology of testis and epididymis changes,and caused a significant loss of multinucleated giant cells,desquamation of germ cells in destructive seminiferous tubules,and degenerative Leydig cells,further destroying the production of sperm.After administration G-Rg3(5 and 10 mg/kg/day)for 2 weeks,the spermatogenic-related indexes of testosterone levels and superoxide dismutase(SOD)activity,glutathione(GSH)content significantly(p<0.01)increase compared with the HS group.Moreover,G-Rg3 treatment effectively ameliorated the production of malondialdehyde(MDA)(p<0.05 or p<0.01).Importantly,G-Rg3 exhibited the protective potential against HS-induced injury not only suppressing the protein levels of heme oxygenase-1(HO-1),hypoxia-inducible factor-1α(HIF-1α),and heat shock protein 70(HSP70)but also modulating the Bcl-2 family(p<0.01 or p<0.001)and activation of mitogen-activated protein kinase(MAPK)signaling pathways(p<0.01).For most of the parameters tested,the HS+G-Rg3(10 mg/kg)group exhibited potent effects compared with those exhibited by the low dose(5 mg/kg)group.In conclusion,the present study demonstrated that G-Rg3 exerted protective effects against HS-induced testicular dysfunction via inhibiting the MAPK-mediated oxidative stress and apoptosis in mice.

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction

Aim： To detect the anti-follicle-stimulating hormone （FSH） antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. Methods： The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay （ELISA）. The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling （HOS） test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy （TEM）. Results： The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa （swollen） was 45.1% ±3.5% in the FSH antibody-positive group and 59.1% ± 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti- FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. Conclusion： These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

Age-dependent expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis

Aim： To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis during postnatal development. Methods： The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results： （1） In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. （2） In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. （3） The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion： The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.

Effects of Local Testicular Heating on Bcl-2 and Bax Protein Expression in Spermatogenic Cells in Rats

Objective To investigate the expression of Bcl-2 and Bax in spermatogenic cells induced by local testicular heating in rats Methods Forty adult male SD rats were divided into experimental group and control group at random. According to day 0.5, 1, 3 and 6 after local testicular heating, each group was divided into 4 subgroups： experimental subgroup （n=6） and control subgroup （n=4）. The expression of Bcl-2 and Bax in the spermatogenic cells was detected on day 0.5, 1, 3 and 6 after heat exposure by using immunohistochemistry. Results Compared with control groups, the ratio of positive cells and content of Bcl-2 positive cells significantly decreased in all experimental subgroups （P〈0.01）. The content of Bax positive cells increased in all experimental subgroups （P〈0.01）, the ratio of positive cells which had no significant difference （P〉0.05） except 6 d group decreased （P〈0.01 ）. Redistribution of Bax from a cytoplasmic to perinuclear or nuclear localization could be observed after heating. Conclusions Expression of Bcl-2 would decrease and Bax would increase with redistribution in spermatogenic cells in rats after heating. The change of Bcl-2 and Bax expression in spermatogenic cells would be correlated with the spermatogenic cell apoptosis induced by heating.

Study on the Regulation of Bcl-2 Gene on Rat Spermatogenic Cells Apoptosis in Transcription Level

Objective To detect the change of Bcl 2 gene expression in the apopototic process of spermatogenic cells in rat with vasoligation and vasostomy, and to find out the relationship between the transcription of Bcl 2 and the apoptosis of spermatognic cells Materials & Methods Sixty adult male Sprague Dawley rats in 3 groups were operated with vasoligation and vasostomy. Then hybridization in situ with hypersensitive Bcl 2 RNA probe was used to detect the change of Bcl 2 mRNA. Results The transcription of Bcl 2 gene in spermatogenic cells was obviously inhibited in the vasoligation group compared with that in the control group (P<0.05), and the transcription in the vasostomy group showed no difference from that of the control group. Conclusion Bcl 2 gene has an anti apoptotic effect in rats with vasostomy, and there was a transcriptional regulation of Bcl 2 gene in rat spermatogenic cell during the period of pre vasoligation to post vasoligation and to post vasosotomy.

A Study on Location of Ureaplasma Urealyticum in Spermatogenic Cells

A study on the location of Ureaplasma Urealyticum (UU) in spermatogenic cells in seminal fluid from infertile males was made using rabbit anti-UU antibodies labelled with horse peroxidase (HRP). The results showed that the positive rates of UU in spermatogenic cells were 39.8% (113/284) and 42.2% (35/83)in the infertile males and the males whose wives had history of habitual abortion, respectively. The positive rates of UU were 45.8% (38/83) and 31.6% (30/95)in the patients with positive and negative anti-sperm antibodies, respectively. A high positive rate of UU (73.3%) was found in the infertile males with oligospermia (<20×109/L). The medicines of tetracyclines, such as minomycin, could effectively inhibit the growth of UU, thus converting the UU-positive into negative and improving the patients'fertility.

Cuproptosis mediates copper-induced testicular spermatogenic cell death

Cuproptosis,a novel mechanism of programmed cell death,has not been fully explored in the context of spermatogenic cells.Thisstudy investigated the potential involvement of cuproptosis in spermatogenic cell death using a mouse model of copper overload.Sixty male Institute of Cancer Research(ICR)mice were randomly divided into four groups that received daily oral gavage withsodium chloride(control)or copper sulfate(CuSO_(4))at 50 mg kg^(−1),100 mg kg^(−1),or 200 mg kg^(−1),for 42 consecutive days.Micesubjected to copper overload exhibited a disruption in copper homeostasis.Additionally,significant upregulated expression of keycuproptosis factors was accompanied by a significant rise in the rates of testicular tissue cell apoptosis.Immunohistochemicalanalysis revealed the presence of ferredoxin 1(Fdx1)in Sertoli cells,Leydig cells,and spermatogenic cells at various stages oftesticular development,and the Fdx1-positive staining area was significantly increased in copper-overloaded mice.Mitochondrialdysfunction and decreased adenosine triphosphate levels were also observed,further implicating mitochondrial damage undercuproptosis.Further analyses revealed pathological lesions and blood−testis barrier destruction in the testicular tissue,accompaniedby decreased sperm concentration and motility,in copper-overloaded mice.In summary,our results indicate that copper-overloadedmice exhibit copper homeostasis disorder in the testicular tissue and that cuproptosis participates in spermatogenic cell death.These findings provide novel insights into the pathogenic mechanisms underlying spermatogenic cell death and provide initialexperimental evidence for the occurrence of cuproptosis in the testis.

Urokinase Receptor Gene Expression Inhibited by siRNA Cocktails in Co-culture of Spermatogenic Cells with Sertoli Cells from 3-Week-Old Rat

Objective To study the functions of urokinase receptor （uPAR） during spermatogenesis by adding targeted siRNA cocktails to the co-culture of Sertoli cell with spermatogenic cells. Methods The seminiferous tubule samples from SD rats aged 20-23 d were digested with collagenase for 15-20 rain, then were cut into smaU fragments. Tubular fragments were digested with collagenase again for 5 -10 min, then gently resuspended in F12/ DMEM supplemented with vitamins and hormones. Cell samples were seeded in plate and cultured at 32 ℃ for 2 weeks. After 48 h, siRNA cocktails at a concentration of 15 nmol/L or 30 nmol/L were introduced into this cocultured Sertoli/spermatogenic cells. After then for 48 h, expression of uPAR mRNA was analyzed by real-time RT-PCR. Results Some spermatogenic cells took place morphologic changes and generated an obvious flagellum. The expression levels of uPAR mRNA silencing with siRNA cocktails at a concentration of 15 nmol/L or 30 nmol/L were significantly lower than those in nontransfected group （P〈0.05）; the inhibition rate was 63.5% or 76.7%, respectively. Conclusion The meiosis and spermiogenesis could take place in this culture system, and the siRNA cocktails can effectively inhibit the gene expression of uPAR in co-cultured Sertoli/spermatogenic cells.

The association of stromal antigen 3(STAG3) sequence variations with spermatogenic impairment in the male Korean population

The stromal antigen 3(STAG3)gene,encoding a meiosis-specific cohesin component,is a strong candidate for causing male infertility,but little is known about this gene so far.We identified STAG3 in patients with nonobstructive azoospermia(NOA)and normozoospermia in the Korean population.The coding regions and their intron boundaries of STAG3 were identified in 120 Korean men with spermatogenic impairments and 245 normal controls by using direct sequencing and haplotype analysis.A total of 30 sequence variations were identified in this study.Of the total,seven were exonic variants,18 were intronic variants,one was in the 5’-UTR,and four were in the 3'-UTR.Pathogenic variations that directly caused NOA were not identified.However,two variants,c.3669+35C>G(rs1727130)and+198A>T(rs1052482),showed significant differences in the frequency between the patient and control groups(P=0.021,odds ratio[OR]:1.79,95%confidence interval[Cl]:1.098-2.918)and were tightly linked in the linkage disequilibrium(LD)block.When pmir-rs1052482A was cotransfected with miR-3162-5p,there was a substantial decrease in luciferase activity,compared with pmir-rs1052482T.This result suggests that rsl052482 was located within a binding site of miR-3162-5p in the STAG33'-UTR,and the minor allele,the rs1052482T polymorphism,might offset inhibition by miR-3162-5p.We are the first to identify a total of 30 single-nucleotide variations(SNVs)of STAG3 gene in the Korean population.We found that two SNVs(rs1727130 and rsl052482)located in the 3'-UTR region may be associated with the NOA phenotype.Our findings contribute to understanding male infertility with spermatogenic impairment.

Bioinformatic identification of key genes and molecular pathways in the spermatogenic process of cryptorchidism

This study aims to determine key genes and pathways that could play important roles in the spermatogenic process of patients with cryptorchidism.The gene expression profile data of GSE25518 was obtained from the Gene Expression Omnibus(GEO)database.Microarray data were analyzed using BRB-Array Tools to identify differentially expressed genes(DEGs)between high azoospermia risk(HAZR)patients and controls.In addition,other analytical methods were deployed,including hierarchical clustering analysis,class comparison between patients with HAZR and the normal control group,gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,and the construction of a proteineprotein interaction(PPI)network.In total,1015 upregulated genes and 1650 downregulated genes were identified.GO and KEGG analysis revealed enrichment in terms of changes in the endoplasmic reticulum cellular component and the endoplasmic reticulum protein synthetic process in the HAZR group.Furthermore,the arachidonic acid pathway and mTOR pathway were also identified as important pathways,while RICTOR and GPX8 were indentified as key genes involved in the spermatogenic process of patients with cryptorchidism.In present study,we found that changes in the synthesis of endoplasmic reticulum proteins,arachidonic acid and the mTOR pathway are important in the incidence and spermatogenic process of cryptorchidism.GPX8 and RICTOR were also identified as key genes associated with cryptorchidism.Collectively,these data may provide novel clues with which to explore the precise etiology and mechanism underlying cryptorchidism and cryptorchidism-induced human infertility.

The association between the two more common genetic causes of spermatogenic failure:a 7-year retrospective study

Chromosomal abnormalities and Y chromosome microdeletions are considered to be the two more common genetic causes of spermatogenic failure.However,the relati on ship between chromosomal aberrations and Y chromosome microdeletio ns is still un clear.This study was to investigate the incidenee and characteristics of chromosomal aberrations and Y chromosome microdeletions in infertile men,and to explore whether there was a correlation between the two genetic defects of spermatogenic failure.A 7-year retrospective study was conducted on 5465 infertile men with nonobstructive azoospermia or oligozoospermia.Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding techniques.Y chromosome microdeletions were screened by multiplex PCR amplification with six specific sequence-tagged site(STS)markers.Among the 5465 infertile men analyzed,371(6.8%)had Y chromosome microdeletions and the prevalence of microdeletions in azoospermia was 10.5%(259/2474)and in severe oligozoospermia was 6.3%(107/1705).A total of 4003(73.2%)infertile men underwent karyotyping;370(9.2%)had chromosomal abnormalities and 222(5.5%)had chromosomal polymorphisms.Karyotype analysis was performed on 272(73.3%)patients with Y chromosome microdeletions and 77(28.3%)had chromosomal aberrations,all of which involved sex chromosomes but not autosomes.There was a sign ifica nt d iff ere nee in the frequency of chromosomal abno rmalities betwee n men with and without Y chromosome microdeletions(P<0.05).

Network Pharmacology-based Strategy for Predicting Active Ingredients and Potential Targets of Cuscutae semen and Lycii fructus in Spermatogenic Dysfunction

Background:Traditional Chinese Medicine(TCM)Cuscutae semen(SC,Tusizi from Cuscuta chinensis Lam.)and Lycii fructus(FL,Gouqi from Lycium barbarum L.),are also used as a herb pair(SC-FL)to treat various ailments,including spermatogenic dysfunction(SD),a disease responsible for low fertility in males.Objective:Herein,we will further determine the bioactive components,component targets and partial molecular mechanisms of the herb pair for the treatment of SD.Methods:We employed the traditional Chinese medicine systems pharmacology database in combination with Ultra high performance liquid chromatography(UHPLC)analysis to analyze the active ingredients of the SC-FL herbal pair and explore its possible targets and underlying mechanism in treatment of spermatogenic dysfunction(SD).Moreover,we used a Sprague-Dawley rat model,generated by using glucosides of Tripterygium wilfordii(GTW),to evaluate the effect of SC-FL and the underlying mechanism on SD and reliability of certain key targets and pathways obtained from the pharmacology analysis.Results:We identified 56 active ingredients in SC-LF affecting 41 overlapping gene signals that influenced SD treatment outcomes.262 Gene Ontology(GO)terms and 170 pathways were yielded under analyses of Gene Ontology,Kyoto Encyclopedia of Gene and Genome pathway from which we predicted that cell proliferation and apoptosis were the primary biological processes(BP)involved in the treatment of SD by SC-LF.The results showed that SC-FL treatment significantly improved the testicular organ coefficient along with sperm count and motility,while reduced testicular damage and testicular tissue cell apoptosis in SD model.Mechanistically,SC-FL significantly upregulated the expression levels of anti-apoptotic proteins AR and BCL2 and downregulated those of pro-apoptotic proteins BAD,BAX,cleaved caspase 3,and CASP3.Conclusion:Collectively,these results indicated that SC-FL elicited a protective effect by potentially regulating apoptosis,thus suggesting that it will represent an effective reagent for male infertility.

Expert recommendations on data collection and annotation of two dimensional ultrasound images in azoospermic males for evaluation of testicular spermatogenic function in intelligent medicine

Testicular two-dimensional ultrasound is a testing modality that is often used to evaluate azoospermia and other related diseases.With the continuous development of deep learning in recent years,the combination of deep learning and testicular ultrasound appears unstoppable despite a lack of relevant standards.One of the major problems associated with the digitization of ultrasound images is the uneven quality of data however,and a standardized data source and acquisition process has not yet been developed.Such a standard could fill the current gap,and establish acquisition criteria for ultrasound images of testes during the male reproductive period,including grayscale ultrasound,shear wave elastography,and contrast-enhanced ultrasound.By following these guidelines the quality of testicular ultrasound images would be improved and standardized,which would lay a solid foundation for the standardization of testicular ultrasound images,and assist automated evaluation of testicular spermatogenic function of whole testis in azoospermic males.