Complete Global Total Electron Content Map Dataset based on a Video Imputation Algorithm VISTA.

Ionospheric total electron content (TEC) derived from multi-frequency Global Navigation Satellite System (GNSS) signals and the relevant products have become one of the most utilized parameters in the space weather and ionospheric research community. However, there are a couple of challenges in using the global TEC map data including large data gaps over oceans and the potential of losing meso-scale ionospheric structures when applying traditional reconstruction and smoothing algorithms. In this paper, we describe and release a global TEC map database, constructed and completed based on the Madrigal TEC database with a novel video imputation algorithm called VISTA (Video Imputation with SoftImpute, Temporal smoothing and Auxiliary data). The complete TEC maps reveal important large-scale TEC structures and preserve the observed meso-scale structures. Basic ideas and the pipeline of the video imputation algorithm are introduced briefly, followed by discussions on the computational costs and fine tuning of the adopted algorithm. Discussions on potential usages of the complete TEC database are given, together with a concrete example of applying this database.

Global Driving of Auroral Precipitation: 1. Balance of Sources.

The accurate determination of auroral precipitation in global models has remained a daunting and rather inexplicable obstacle. Understanding the calculation and balance of multiple sources that constitute the aurora, and their eventual conversion into ionospheric electrical conductance, is critical for improved prediction of space weather events. In this study, we present a semi-physical global modeling approach that characterizes contributions by four types of precipitation-monoenergetic, broadband, electron, and ion diffuse-to ionospheric electrodynamics. The model uses a combination of adiabatic kinetic theory and loss parameters derived from historical energy flux patterns to estimate auroral precipitation from magnetohydrodynamic (MHD) quantities. It then converts them into ionospheric conductance that is used to compute the ionospheric feedback to the magnetosphere. The model has been employed to simulate the 5-7 April 2010 Galaxy15 space weather event. Comparison of auroral fluxes show good agreement with observational data sets like NOAA-DMSP and OVATION Prime. The study shows a dominant contribution by electron diffuse precipitation, accounting for ∼74% of the auroral energy flux. However, contributions by monoenergetic and broadband sources dominate during times of active upstream solar conditions, providing for up to 61% of the total hemispheric power. The study also finds a greater role played by broadband precipitation in ionospheric electrodynamics which accounts for ∼31% of the Pedersen conductance.

Exosomes released from Sertoli cells contribute to the survival of Leydig cells through CCL20 in rats.

Reciprocal communication between Sertoli and Leydig cells occurs in the testes; however, the detailed mechanisms involved are not completely understood. Exosomes can communicate within neighboring or distant cells to regulate cell function. Our aim was to determine whether exosomes released from Sertoli cells can regulate the survival of Leydig cells. We found that exosomes released from rat primary Sertoli cells could be internalized by Leydig cells in vitro, and promote the survival of Leydig cells, as assessed by optical density at 450 nm, compared to untreated control (mean ± SD: 0.95 ± 0.04 vs 0.79 ± 0.03, P < 0.05). When the exosomes were injected into the interstitial area of rat testis, they could also be internalized by Leydig cells in vivo. To investigate if exosomes released from Sertoli cells can reach Leydig cells in vivo, exosomes were injected into the efferent duct, from where they entered the interstitial space from seminiferous tubules, which indicated that they may cross the blood-testis barrier (BTB). Further in vitro studies found that exosomes released from Sertoli cells significantly increased CC-chemokine ligand 20 (Ccl20) mRNA (mean ± SD: 2.79 ± 0.08 vs 0.98 ± 0.04, P < 0.01) and protein (mean ± SD: 1.08 ± 0.06 vs 0.53 ± 0.05 ng/ml, P < 0.01) levels in Leydig cells, compared to the untreated Leydig cells. CCL20 promoted the phosphorylation of AKT (protein kinase B) in Leydig cells, compared to untreated control (mean ± SD: 0.074 ± 0.002 vs 0.051 ± 0.002, P < 0.01). In conclusion, our results demonstrated that exosomes released by Sertoli cells may cross the BTB and promote the survival of Leydig cells. The findings may add new evidence for Sertoli-Leydig cell communication.

Comparison of intracytoplasmic sperm injection (ICSI) outcomes in infertile men with spermatogenic impairment of differing severity.

The extent of spermatogenic impairment on intracytoplasmic sperm injection (ICSI) outcomes and the risk of major birth defects have been little assessed. In this study, we evaluated the relationship between various spermatogenic conditions, sperm origin on ICSI outcomes, and major birth defects. A total of 934 infertile men attending the Center for Reproductive Medicine of Ren Ji Hospital (Shanghai, China) were classified into six groups: nonobstructive azoospermia (NOA; n = 84), extremely severe oligozoospermia (esOZ; n = 163), severe oligozoospermia (sOZ, n = 174), mild oligozoospermia (mOZ; n = 148), obstructive azoospermia (OAZ; n = 155), and normozoospermia (NZ; n = 210). Rates of fertilization, embryo cleavage, high-quality embryos, implantation, biochemical and clinical pregnancies, abortion, delivery, newborns, as well as major birth malformations, and other newborn outcomes were analyzed and compared among groups. The NOA group showed a statistically lower fertilization rate (68.2% vs esOZ 77.3%, sOZ 78.0%, mOZ 73.8%, OAZ 76.6%, and NZ 79.3%, all P < 0.05), but a significantly higher implantation rate (37.8%) than the groups esOZ (30.1%), sOZ (30.4%), mOZ (32.6%), and OAZ (31.0%) (all P < 0.05), which was similar to that of Group NZ (38.4%). However, there were no statistically significant differences in rates of embryo cleavage, high-quality embryos, biochemical and clinical pregnancies, abortions, deliveries, major birth malformations, and other newborn outcomes in the six groups. The results showed that NOA only negatively affects some embryological outcomes such as fertilization rate. There was no evidence of differences in other embryological and clinical outcomes with respect to sperm source or spermatogenic status. Spermatogenic failure and sperm origins do not impinge on the clinical outcomes in ICSI treatment.

Seminal exosomal miR-210-3p as a potential marker of Sertoli cell damage in Varicocele.

BACKGROUND: Varicocoele-associated stressors, such as hypoxia and heat, can damage cell function and viability, and some exosomal biomarkers released from impaired cells may reflect the cell status in testis. OBJECTIVES: To find if seminal exosomal microRNAs can reflect the Sertoli cell function in varicocoele. MATERIALS AND METHODS: Experimental left varicocoele rat model was established (n = 24), and patients with different grades of varicocoele (n = 104) were enrolled. Primary rat Sertoli cells were isolated with enzymatic hydrolysis. Exosomes were isolated from primary rat Sertoli cells, rat epididymis tissue, and human seminal plasma with polymer-based precipitation method. Exosomal microRNAs were quantified with qPCR. Inhibin-B was detected with enzyme immunoassay. The correlation analysis between microRNA and inhibin-B was evaluated with Spearman's correlation. RESULTS: We screened 12 previously reported hypoxia-responsive microRNAs in the primary rat Sertoli cells and found that 4 exosomal microRNAs increased significantly in response to in vitro hypoxia treatment (P < .05). Of the 4 microRNAs, only miR-210-3p was upregulated in the rats with experimental varicocoele (P < .01). In the patients with varicocoele, we found that seminal exosomal miR-210-3p significantly increased in patients with grade II and III varicocoele (P < .01), and miR-210-3p negatively correlated with sperm count (P < .01) and seminal inhibin-B expression (r = -0.39, P < .01). For the 30 patients with microsurgical varicocelectomy, the operation notably decreased miR-210-3p (P < .01). DISCUSSION AND CONCLUSION: Seminal exosomal miR-210-3p may be a novel, sensitive, and non-invasive biomarker of Sertoli cell damage in varicocoele.

Nitrifying biomass can retain its acclimation to 2,4,6-trichlorophenol.

Many municipal wastewater treatment plants in China receive industrial wastewater that contains inhibitory organic chemicals, such as chlorinated phenols. For the common aerobic biological treatment, nitrification is a key step, but nitrifying bacteria are notably sensitive to inhibition by chlorinated phenols. In this work, normal activated sludge (containing nitrifying biomass) was acclimated to 2,4,6-trichlorophenol (TCP). The acclimated biomass had more than 2-fold faster nitrification kinetics than normal biomass when exposed to TCP, and it also achieved effective TCP removal in parallel. When suddenly exposed to TCP after as much as two months without TCP input, the acclimated nitrifying biomass retained effective nitrification and TCP biodegradation: The nitrification rate and TCP removal rate were 0.325 mM/h and 0.049 mM/h for the acclimated biomass, compared to only 0.165 mM/h and 0.001 mM/h for normal biomass. Resistance to TCP inhibition also was retained for 5 generations of sub-culturing without TCP exposure. High-throughput sequencing confirmed that the acclimated biomass contained nitrifying bacteria and heterotrophic bacteria capable of degrading TCP, although the key genera changed during sub-culturing.

Bioavailable electron donors from ultrasound-treated biomass for stimulating denitrification.

Finding low-cost electron donors to drive denitrification is an important target for many municipal wastewater treatment plants (MWTPs). Excess sludge (biomass) potentially is a low-cost electron donor generated internally to the MWTP, but it has to be made more bioavailable. Aerobic and anoxic biomasses were treated with ultrasound, and their supernatants were used as electron donors for stimulating denitrification. The supernatant from ultrasound-treated anoxic biomass achieved 54% faster nitrate-N removal than did supernatant from the treated aerobic biomass, and the supernatant of untreated biomass was ineffective as an electron donor. UV illumination of the supernatants further enhanced the rates, with increments of 19% and 14%, respectively for the aerobic and anoxic supernatants. Sodium acetate at a range of initial concentrations was compared as a readily bioavailable electron donor to gauge the acceleration impact of the supernatants as equivalent bioavailable chemical oxygen demand (COD). The total chemical oxygen demand (TCOD) of the supernatant harvested from anoxic biomass without UV illumination was 76% bioavailable, while its bioavailable TCOD was 78% after UV illumination. For the supernatant from the aerobic biomass, the bioavailable fractions were, respectively, 56% and 58% without and with UV illumination. The greatest impact for converting excess biomass into a source of bioavailable electron donor to drive denitrification came from ultrasound treatment of the biomass, which disrupted the biomass to form particulate chemical oxygen demand (PCOD) that was bioavailable. PCOD was at least 51% bioavailable, and it contributed no less than 82% of the bioavailable COD.

Comparison and outcomes of nonobstructive azoospermia patients with different etiology undergoing MicroTESE and ICSI treatments.

BACKGROUND: The purpose of the study was to compare clinical, laboratory, histological features, microdissection testicular sperm extraction (MicroTESE) and intracytoplasmic sperm injection (ICSI) treatment outcomes of nonobstructive azoospermia (NOA) patients of various etiologies, and to investigate ICSI outcomes using fresh and frozen thawed sperms from MicroTESE, so to explore an optimal MicroTESE-ICSI procedure for NOA couples. METHODS: A retrospective analysis was made in 595 NOA patients undergoing MicroTESE from January 2013 to December 2017. The men were classified into six groups based on etiology. Patients' age, history, hormone profile, testis volume, testicular histology, sperm retrieval, fertile and pregnancy outcomes of ICSI were included for analysis. RESULTS: A total of 595 NOA patients were included in this study, with 446 (75.0%) were idiopathic NOA, 66 (11.1%) were Klinefelter syndrome (KS), 34 (5.7%) with microdeletion of the AZFc, 33 (5.5%) cases had the history of cryptorchidism, 13 (2.2%) had a history of mumps orchitis, and 3 (0.5%) cases underwent chemotherapy. The overall sperm retrieval rate (SRR) was 40.3% (240/595), SRR of the cryptorchidism (84.8%, 28/33) and mumps orchitis (84.6%, 11/13) groups were much higher than that of other groups, the SRR of idiopathic group was the lowest (31.8%, 142/446). One hundred and ninety-eight ICSI cycles utilizing MicroTESE sperm were retrospectively analyzed, including 155 fresh MicroTESE ICSI cycles and 43 frozen-thawed MicroTESE ICSI cycles. Fertilization rate, cleavage rate, and clinical pregnancy rate of fresh sperm group were slightly higher than those in frozen thawed MicroTESE sperm group, but high qualified embryo rate of fresh sperm group was lower than frozen thawed group. The differences were of no statistical meaning. CONCLUSIONS: Etiology may be an effective prognostic factor for SRR in NOA patients. NOA of definite etiology, such as cryptorchidism, has high SRR, while idiopathic NOA, the most common type of NOA, has the lowest SRR. Using of frozen thawed sperm from MicroTESE had similar pregnant outcome to that of fresh sperm, so cryopreservation of testicular sperm seems to be more suitable and of great benefit in these cases and good results can also be expected when oocyte retrieval and ICSI are not performed at the same time.

Acclimation of nitrifying biomass to phenol leads to persistent resistance to inhibition.

It is common that biological wastewater-treatment processes are exposed to inputs of toxic compounds, such as phenolics. Due to their slow growth rate, nitrifying bacteria are most susceptible to inhibition that can lead to loss of nitrification capacity. Here, a microbial community containing nitrifying bacteria was acclimated to phenol, and it developed resistance to phenol inhibition and maintained nitrification activity. For the phenol-acclimated biomass, the NH4+-N removal rates were almost unaffected when it was suddenly exposed to phenol. Heterotrophic synthesis and nitrification rates contributed 76% and 24% of the total NH4+-N removal respectively during phenol removal, but the nitrification rate increased significantly once phenol was removed and mineralized. In contrast, the NH4+-N removal rates decreased sharply for normal (unacclimated) nitrifying biomass when it was exposed to phenol. The phenol-acclimated biomass retained its resistance to phenol inhibition for at least two months after acclimation, and addition of the phenol-acclimated biomass to the normal biomass conferred resistance to phenol inhibition. Community analysis of the phenol-acclimated biomass showed an increase in families known to contain strains able to biodegrade phenolics. Taken together, the results indicate that the main impact of phenol acclimation was enrichment of phenol-biodegrading bacteria, which allowed rapid removal and mineralization of phenol and, consequently, alleviation of phenol's inhibition of nitrification.

Simultaneous anaerobic and aerobic transformations of nitrobenzene.

Aerobic biodegradation of nitrobenzene (NB) produces nitrophenol (NP), which has stronger toxicity than NB. Anaerobic biodegradation of NB produces aniline (AN), which has weaker toxicity, but is a dead-end product in anaerobic conditions. Accumulation of AN should be overcome by coupling anaerobic and aerobic transformations: NB is transformed to AN in an anaerobic zone of the bioreactor, and AN is then transformed in an aerobic zone. A vertical baffled bioreactor (VBBR) was employed for NB biodegradation with a controlled dissolved oxygen (DO) concentration. NB biodegradation was accelerated by simultaneous anaerobic and aerobic transformations, since AN was biotransformed by a mono-oxygenase reaction. Adding exogenous electron donor (acetate) enhanced NB removals when the DO concentration was ∼0.5 mg/L, because the donor accelerated mono-oxygenations of NB and AN. Coupling anaerobic and aerobic transformations can be a valuable strategy for biodegrading organic compounds that undergo aerobic and anaerobic biotransformations.

A novel micro-straw for cryopreservation of small number of human spermatozoon.

Cryopreservation of few spermatozoa is still a major challenge for male fertility preservation. This study reports use a new micro-straw (LSL straw) for freezing few spermatozoa for intracytoplasmic sperm injection (ICSI). Semen samples from 22 fertile donors were collected, and each semen sample was diluted and mixed with cryoprotectant in a ratio of 1:1, and then frozen using three different straws such as LSL straw (50-100 µl), traditional 0.25 ml and 0.5 ml straws. For freezing, all straws were fumigated with liquid nitrogen, with temperature directly reducing to -130--140°C. Sperm concentration, progressive motility, morphology, acrosome integrity, and DNA fragmentation index were evaluated before and after freezing. After freezing-thawing, LSL straw group had significantly higher percentage of sperm motility than traditional 0.25 ml and 0.5 ml straw groups (38.5% vs 27.4% and 25.6%, P < 0.003). Sperm motility and acrosomal integrity after freezing-thawing were significantly lower than that of before freezing. However, there was no significant difference in morphology, acrosome, and DNA integrity between the three types of straws (P > 0.05). As LSL straws were thinner and hold very small volume, the freezing rate of LSL straw was obviously faster than 0.25 ml straw and 0.5 ml straws. In conclusion, LSL micro-straws may be useful to store few motile spermatozoa with good recovery of motility for patients undergoing ICSI treatment.

Association of a TDRD1 variant with spermatogenic failure susceptibility in the Han Chinese.

PURPOSE: Piwi-interacting RNAs (piRNAs) are a broad group of noncoding small RNAs that have important biological functions in germline cells and can maintain genome integrity via silencing of retrotransposons. In this study, we aimed to explore the associations between genetic variants of important genes involved in piRNA biogenesis and male infertility with spermatogenic impairment. METHODS: To this end, five single-nucleotide polymorphisms (SNPs) in the ASZ1, PIWIL1, TDRD1, and TDRD9 genes were genotyped by TaqMan allelic discrimination assays in 342 cases of nonobstructive azoospermia (NOA) and 493 controls. RESULTS: The SNP rs77559927 in TDRD1 was associated with a reduced risk of spermatogenic impairment. The genotypes TC and TC + CC showed odds ratios and 95 % confidence intervals of 0.73 (0.55-0.98, P = 0.034) and 0.73 (0.56-0.97, P = 0.030), respectively, in patients with NOA compared with those in the controls. CONCLUSION: Thus, our results provided the first epidemiological evidence supporting the involvement of TDRD1 genetic polymorphisms in piRNA processing genes in determining the risk of spermatogenic impairment in a Han Chinese population.

VEGF/VEGFR2 Signaling Regulates Germ Cell Proliferation in vitro and Promotes Mouse Testicular Regeneration in vivo.

Vascular endothelial growth factor (VEGF) plays fundamental roles in testicular development; however, its function on testicular regeneration remains unknown. The objective of this study was to explore the roles VEGF/VEGFR2 signaling plays in mouse germ cells and in mouse testicular regeneration. VEGF and the VEGFR2 antagonist SU5416 were added to culture medium to evaluate their effects on spermatogonial stem cell line (C18-4 cells) proliferation. Testicular cells obtained from newborn male ICR mice were grafted into the dorsal region of male BALB/c nude mice. VEGF and SU5416 were injected into the graft sites to assess the effects of the VEGF and VEGFR2 signaling pathways on testicular reconstitution. The grafts were analyzed after 8 weeks. We found that VEGF promoted C18-4 proliferation in vitro, indicating its role in germ cell survival. HE staining revealed that seminiferous tubules were reconstituted and male germ cells from spermatogonia to spermatids could be observed in testis-like tissues 8 weeks after grafting. A few advantaged male germ cells, including spermatocytes and spermatids, were found in SU5416-treated grafts. Moreover, VEGF enhanced the expression of genes specific for male germ cells and vascularization in 8-week grafts, whereas SU5416 decreased the expression of these genes. SU5416-treated grafts had a lower expression of MVH and CD31, indicating that blockade of VEGF/VEGFR2 signaling reduces the efficiency of seminiferous tubule reconstitution. Collectively, these data suggest that VEGF/VEGFR2 signaling regulates germ cell proliferation and promotes testicular regeneration via direct action on germ cells and the enhancement of vascularization.

Endothelial nitric oxide synthase (eNOS) T-786C, 4a4b, and G894T polymorphisms and male infertility: study for idiopathic asthenozoospermia and meta-analysis.

Recent studies on the eNOS gene and male infertility show that expression of eNOS regulates normal spermatogenesis in the testis, and the eNOS gene variants (T-786C, 4a4b, and G894T) are potentially involved in impairment of spermatogenesis and sperm function. Thus, we conducted this association and meta-analysis study to further validate whether variants of those three loci affected the risk of idiopathic asthenozoospermia (AZS) and male infertility. Approximately 340 Chinese idiopathic AZS patients and 342 healthy men were included for this case-control study, genotyped by gel electrophoresis analysis or direct sequencing of PCR products. The eNOS mRNA isolated from the semen of patients was further examined by quantitative real-time PCR. Also, a meta-analysis of association between eNOS gene polymorphisms and male infertility was performed. A significant association was identified on allelic level between 4a4b variant and AZS in our study (chi-squared = 7.53, corrected P = 0.018, odds ratio (OR) = 1.808), while there were no significant difference of T-786C and G894T for asthenozoospermia in both genotype and allele distributions. In addition, expression of eNOS was up-regulated in patients compared with controls (about 2.4-fold, P < 0.001). Furthermore, the results of the meta-analysis support the conclusion that the T-786C and 4a4b loci were associated with male infertility in both Asian and Caucasian populations. Our study provides genetic evidence for the eNOS gene being a risk factor for idiopathic AZS and male infertility. Considering genetic differences among populations and complex pathogenesis of male infertility, more validating studies using independent samples are suggested in the future.

Mesenchymal stem cells overexpressing Ihh promote bone repair.

BACKGROUND: Indian hedgehog (Ihh) signaling pathway is known to play key roles in various aspects of normal endochondral bone development. This study tested the potential roles of high Ihh signaling in the context of injury-induced bone regeneration. METHODS: A rabbit tibia defect model was established to test the effects of the implant of Ihh/mesenchymal stem cells (MSCs)/scaffold complex. Computed tomography (CT), gross observation, and standard histological and immunohistological techniques were used to evaluate the effectiveness of the treatment. In vitro studies with MSCs and C3H10T1/2 cells were also employed to further understand the cellular and molecular mechanisms. RESULTS: We found that the implanted Ihh/MSCs/scaffold complex promoted bone repair. Consistently, in vitro study found that Ihh induced the upregulation of chondrocytic, osteogenic, and vascular cell markers, both in C3H10T1/2 cells and MSCs. CONCLUSIONS: Our study has demonstrated that high Ihh signaling in a complex with MSCs enhanced bone regeneration effectively in a clinically relevant acute injury model. Even though the exact underlying mechanisms are still far from clear, our primary data suggested that enhanced chondrogenesis, osteogenesis, and angiogenesis of MSCs at least partially contribute to the process. This study not only has implications for basic research of MSCs and Ihh signaling pathway but also points to the possibility of direct application of this specific paradigm to clinical bone repair.

Association study between polymorphisms of PRMT6, PEX10, SOX5, and nonobstructive azoospermia in the Han Chinese population.

A previous genomewide association study of nonobstructive azoospermia (NOA) in the Han Chinese population identified three risk loci (rs12097821, rs2477686, and rs10842262) and provided strong evidence for a genetic influence in male infertility. However, recently, a follow-up study of these single nucleotide polymorphism (SNP) loci in the Japanese population showed that none of them were significantly associated with NOA. Therefore, we conducted an association study, consisting of 550 NOA cases and 555 normal controls, to further validate whether the risk of those three SNPs still existed in an independent Han Chinese male population. The association studies did not support the association of rs12097821 and rs2477686 with NOA for both genotype and allele distributions, but rs10842262 in the SOX5 gene was significantly associated with NOA (chi square = 9.31; P = 0.0095 and chi square = 9.27; P = 0.0023, respectively). Our study provides genetic evidence for SOX5 polymorphism in NOA, contributing to predicting males at high risk of NOA in Han Chinese population. Considering genetic differences among populations, future validating studies in independent samples are suggested.

Ectopic expression of Sonic Hedgehog in a cryptorchid man with azoospermia: a case report.

A 30-year-old man presented with a left undescended testis, right testicular deficiency and azoospermia. Testicular biopsy revealed an absence of spermatocytes and increased numbers of Leydig cells in the undescended testis. Additional comparative analyses were undertaken to explore Sonic Hedgehog (Shh) immunostaining in the testis of juvenile and adult mice, in the testis of the patient with cryptorchidism, and in archival testicular tissue from a patient with obstructive azoospermia and a patient with prostate cancer. Shh immunostaining was demonstrated in spermatocytes in juvenile and adult mouse testis and in the patients with obstructive azoospermia and prostate cancer, suggesting that Shh signalling is involved in normal spermatogenesis. In the patient with cryptorchidism, Shh immunostaining was localized to the Leydig cells, which suggests that Shh might be involved in the abnormal expansion of the Leydig cell population in the testis. These preliminary data on the appearance of Shh protein during normal spermatogenesis might provide the basis for further investigations to clarify the role of Shh signalling in spermatogenesis during normal and pathogenic testis development.

[Pre-freezing equilibration improves post-thaw human semen parameters].

OBJECTIVE: To investigate the effect of pre-freezing equilibration on the cryo-survival of human sperm and to optimize the protocol of direct fumigation for the freeze-thawing of human sperm. METHODS: We collected 50 semen samples from healthy donors, each subjected to cryopreservation with 3 different methods: non-equilibration freezing (Group A), 10-min equilibration at room temperature before freezing (Group B), and 10-min equilibration at 4 degrees C before freezing (Group C). We examined all the post-thaw semen samples by computer-assisted semen analysis for the sperm motility parameters, and detected the sperm vitality and deformity index (SDI). RESULTS: The recovery rate of progressive sperm motility was (61.88 +/- 16.94)% in Group C, remarkably higher than in A ([48.61 +/- 16.44]%) and B ([49.41 +/- 13.77]%) (P < 0.05), but with no significant difference between the latter two. And there were no significant differences in sperm vitality and SDI among the three groups. CONCLUSION: Ten-minute equilibration at 4 degrees C before freezing can evidently improve the progressive motility of sperm in addition to its advantages of easy operation and controllable experimental condition.

[Desert hedgehog regulates the proliferation and differentiation of Leydig cells: an update].

Hedgehog signaling pathway plays an important role in regulating the development of endocrine glands. Desert hedgehog (Dhh) has become a recent focus for its regulation of testis development, especially of Leydig cells. Dhh, as a Sertoli cell product, regulates the proliferation and differentiation of Leydig cell lineage and functions to secrete testosterone through a paracrine signaling mechanism. Testosterone, as the most important sex hormone of the male, plays a critical role in testis development, spermatogenesis and maintenance of normal masculinization. Therefore, normal Dhh signaling pathway ensures normal spermatogenic function. Researches on the Hedgehog signaling pathway in the testis have a potential significance for studying the pathophysiological mechanisms of androgen deficiency and dyszoospermia, as well as for the clinical treatment of these diseases.

[Reconstructed seminiferous tubules from ectopic grafting of testicular cells].

OBJECTIVE: To investigate the ectopic grafts of mouse testicular cells by observing the reconstruction of seminiferous tubules, colonization of spermatogenic cells and spermatogenesis using immunodeficient mice as recipients. METHODS: The testes of newborn male ICR mice were digested to obtain single cell suspension. The cells were then mixed with matrigel and subcutaneously grafted into the dorsal region of the male nude mice. The mice were castrated after the operation and the grafts were dissected from 5 of the nude mice at 4, 6, 8 and 10 weeks, respectively. The success rates of transplantation and the graft diameters were calculated, and the structure of the reconstituted seminiferous tubules, colonization of the germ cells and spermatogenesis were observed by HE staining and immunohistochemistry. RESULTS: All the mice recipients survived after the testicular cell transplantation. Within 10 weeks after the operation, tissue masses could be observed, with the diameter increased from (3.91 +/- 0.71) mm at 4 weeks to (6.69 +/- 0.50) mm. Neovascularization was detected at the surface of the masses and seminiferous tubule structures found in the grafts. The germ cells that developed from spermatogonia to round spermatids were observed, but with no sperm in the tubules. Germ cells, Sertoli cells and Leydig cells were identified by immunochemical detection of Mvh, Gata4 and P450Scc in the grafts at 8 weeks. CONCLUSION: Seminiferous tubules could be ectopically reconstructed from suspension of neonatal mouse testicular cells. Ectopic grafting provided a preferable model for the studies on testis tissue engineering and interactions between testicular cells during testicular development and spermatogenesis.