Nerve growth factor promotes human sperm motility in vitro by increasing the movement distance and the number of A grade spermatozoa

Nerve growth factor (NGF) was first found in the central nervous system and is now well known for its multiple pivotal roles in the nervous system and immune system. However, more and more evidences showed that NGF and its receptors TrkA and p75 were also found in the head and tail of spermatozoa, which indicate the possible effect of NGF on the sperm motility. Nevertheless, the exact role of NGF in the human sperm motility remains unclear until now. In this study, we investigated the effect of NGF on human sperm motility, and the results showed that NGF could promote human sperm motility in vitro by increasing the movement distance and the number of A grade spermatozoa. Further analysis demonstrated that NGF promoted the sperm motility in a dose‐dependent manner in vitro. These results may facilitate the further studies on human fertility and assisted reproduction techniques.     

添加黄芪注射液对人精子线粒体功能的影响

目的 观察添加黄芪注射液对人精子线粒体功能的影响。方法 弱精子症精子与黄芪注射液共孵化,用若丹明(Rh123)和碘化吡啶(PI)染色并用流式细胞技术检测人精子线粒体功能。结果 添加黄芪组线粒体功能良好的活精子数(9.831.65)明显高于对照组(6.500.41),与对照组比较差异有显著性(P<0.05)。添加黄芪组(42.493.73)的线粒体荧光值也明显高于对照组(31.074.49),与空白对照组比较差异有显著性(P<0.05)。结论 体外添加黄芪注射液能增强弱精子症精子线粒体的活性,为黄芪作为体外添加剂提高精子活力提供理论基础。     

Evidence suggesting that germ cells influence the bidirectional secretion of androgen binding protein by the seminiferous epithelium demonstrated by selective impairment of spermatogenesis with busulphan

Androgen binding protein (ABP) was measured in the serum, testes and epididymides of adult rats up to 105 days after the induction of reversible impairment of spermatogenesis by a single injection of busulphan. This treatment decreased testicular and epididymal weights within 7-21 days after treatment, reaching a minimum at 63 days with partial recovery by 105 days. The testicular and epididymal content of sperm was unchanged up to 42 days after busulphan administration, was reduced considerably at 63 days and thereafter increased towards control values. The serum and testicular concentrations of testosterone were normal at all times after treatment, even though serum LH levels were increased at 42 and 63 days. Serum levels of FSH were also increased at 43 and 63 days after treatment. A biphasic pattern in the serum levels of ABP was observed. Concentrations were low up to 43 days post treatment when only the early germ cell types were depleted from the seminiferous epithelium and when the testicular and epididymal contents of ABP were normal. Serum levels of ABP increased as the more mature germ cells were depleted in numbers and the testicular and epididymal contents of ABP declined. It is concluded that bidirectional secretion of ABP into the interstitium (serum) and into the seminiferous tubular lumen by Sertoli cells is influenced considerably by the population of germ cells that are present in the seminiferous epithelium.     

The effects of female age on the outcome of testicular sperm extraction and intracytoplasmic sperm injection in infertile patient with azoospermia

Introduction: Testicular sperm extraction (TESE) is well-defined procedure for surgical sperm retrieval in obstructive and non-obstructive azoospermia. This study was focused on the effectiveness of testicular sperm extraction and intracytoplasmic sperm injection (ICSI) for azoospermic men with different female age subgroups.Materials and methods: A total of 107 men with azoospermia underwent TESE and ICSI treatment. The women were examined in three groups 20–29, 30–34 and 35 years or older. The main outcome in this study was fertilization and pregnancy rates with TESE and ICSI.Results: Spermatozoa were successfully retrieved during 97 of 107 (90.7%) TESE attempts, resulting in the fertilization of 286 of 563 (50.4%) injected metaphase II oocytes. Two hundred and fifty-five of them were transferred (89.8%). The clinical pregnancy rate and ongoing pregnancy rate per embryo transfer were 22.5% and 20.6% respectively. When comparing the fertilization and pregnancy rates, it was observed that women between the ages of 20–29 years had significantly higher pregnancy rates than women over 34 years of age (p < 0.05).Conclusion: The female age is a major factor in determining successful implantation in ICSI.     

NGF promotes mitochondrial function by activating PGC‐1α in TM4 Sertoli cells

Nerve growth factor (NGF), which is required for the survival and differentiation of the nervous system, has been proved to play important roles in male reproductive physiology. Several studies have focused on the roles of NGF in the testes. However, no study has reported on the mechanism of paracrine and autocrine actions of NGF in Sertoli cells. This study showed that NGF stimulated mitochondrial activity and biogenesis in TM4 Sertoli cells. Moreover, our results demonstrated that peroxisome proliferator‐activated receptor‐gamma coactivator‐1α is a possible downstream key target of the NGF signalling pathway. In a 3‐nitropropionic acid cell model, NGF treatment attenuated mitochondrial activity defect and depolarisation. This NGF‐triggered signalling may help in discovering new therapeutic targets for certain male infertility disorders.     

Effect of the age of broodstock males on sperm function during cold storage in the trout (Oncorhynchus mykiss)

The knowledge of sperm quality in the broodstock males of different ages is a prerequisite to identify the reproductive ability of cultivated fish for the hatchery management. Thus, in this work, we analysed sperm function of the semen stored of broodstock males of rainbow trout (Oncorhychus mykiss) in different reproductive ages (2, 3 and 4 years old). Sperm samples of each reproductive age were stored in Storfish^® during 10 days at 4°C, and then, motility, viability, mitochondrial function (MMP), superoxide anion () level and DNA fragmentation (DNA_frag) were assessed. The results demonstrated that sperm function parameters were affected significantly by the age of the males and the time of storage. Motility, viability and MMP significantly decreased, and DNA_frag and level increased with the age increment and the time of storage. In conclusion, sperm quality of 2 and 3 years old were superior to those of 4 years old, based on higher quality of various sperm functions such as motility, viability, MMP, DNA integrity and level during short‐term storage. This information must be considered for optimum utilization of broodstock males in aquaculture.     

Apoptosis precedes detachment of germ cells from the seminiferous epithelium after hormone suppression by short-term oestradiol treatment of rats

Spermatogenesis is a highly synchronized process in which FSH and testosterone are considered the major regulators. Nevertheless, the mechanism by which these hormones act on germ cells is unclear. Cell adhesion has been proved to play an essential role in the regulation of programmed cell death in epithelial cells and it is now known that FSH and testosterone withdrawal results in the triggering of apoptosis as well as germ cell detachment from the seminiferous epithelium. Therefore, it seemed important to investigate whether the triggering of apoptosis in germ cells by experimental hormone suppression occurred as a result of their previous detachment from the epithelium. To achieve this goal, adult male rats were injected with 50 μg oestradiol benzoate for 1, 2, 3, 4, 5 or 10 days to suppress gonadotrophin secretion and thus intratesticular levels of testosterone. Germ cell apoptosis was assessed in testes from these animals by in situ 3' end-labelling of DNA fragments and quantified in seminiferous tubule sections at stages VII–VIII. Serial sections throughout the epididymides from these animals were analysed to search for immature germ cells detached from the epithelium. These cells were scored and quantified in non-consecutive randomly selected epididymal sections. Our data indicate that the triggering of apoptosis in germ cells precedes germ cell detachment, suggesting that detachment of germ cells from the epithelium, occurring after hormone suppression, is not necessary for germ cell apoptosis.     

Proliferation of Sertoli cells during development of the human testis assessed by stereological methods

Sertoli cells were studied using stereological methods in testes obtained from five children who were stillborn, and 31 individuals between 3 months and 40 years of age, who had suffered from sudden, unexpected death. The mean nuclear volume of the Sertoli cells, the numerical density of Sertoli cells, and the total number of Sertoli cells per individual were determined by point- and profile-counting of 0.5 micron sections. The nuclear volume of Sertoli cells increased from a median of 120 microns3 (range 53-130) during the period of 3 months to 10 years to 210 microns3 (170-260) in adults (greater than 25 years). The numerical density of Sertoli cells decreased from a median of 1200 X 10(6)/cm3 (870-1400) during childhood (3 months to 10 years) to 140 X 10(6)/cm3 (110-260) in adults (greater than 25 years). The total number of Sertoli cells per individual increased significantly from a median of 260 X 10(6) (130-520) during the late foetal period to 1500 X 10(6) (850-2900) in individuals from 3 months to 10 years of age. A further increase was found during puberty as the number of Sertoli cells in adults (greater than 25 years) was 3700 X 10(6) (2500-5600). These results indicate that significant qualitative and quantitative changes in the population of Sertoli cells take place after birth.     

Accuracy of the normal sperm morphology value by Sperm Quality Analyzer IIC: comparison with the strict criteria

The aim of the present study is to investigate the accuracy of the normal sperm morphology value by Sperm Quality Analyzer IIC (SQA IIC), which was developed to provide a rapid and low-cost quantitative evaluation of semen quality. Normal sperm morphology was assessed using SQA IIC in comparison with that by the strict criteria in 62 semen samples. Normal sperm morphology value by SQA IIC was based on the studies of three traditional sperm parameters from over 4000 fresh, untreated semen samples, while the strict criteria was based on the method by Kruger et al. The mean +/- SD of percent normal morphology by SQA IIC and the strict criteria were 37.6 +/- 10.9% (range 15-52) and 19.9 +/- 8.2 (range 1-34), respectively. There was a significant correlation of the sperm morphology assessment between the two methods (r=0.454, p < 0.001). Using the cut-off value of >30% normal morphology by SQA IIC, the positive predictive value and the negative predictive value of the 'normal' strict criteria were 79.6% (39/49) and 46.2% (6/13), respectively. These results indicate that SQA IIC might be used as an initial screening test for the evaluation of sperm morphology. However, sperm morphological assessment by the strict criteria should be performed in order to make decisions in planning strategies for the treatment of infertile couples.     

Downregulation of PARVA promotes metastasis by modulating integrin-linked kinase activity and regulating MAPK/ERK and MLC2 signaling in prostate cancer

BackgroundMetastasis is the predominant cause of mortality in prostate cancer (PCa); however, the underlying mechanisms are largely uncharted. Here, we found that Parvin alpha (PARVA) is downregulated in PCa and its loss is associated with clinical metastasis. We further explored the mechanistic basis of this finding.MethodsThe mRNA expression of PARVA was identified by analysis of the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) data sets. Immunohistochemistry (IHC) analysis was performed to evaluate the PARVA expression pattern in 198 PCa tissues, and 36 metastatic lymph node tissues. The function and molecular mechanism by which PARVA affects PCa were investigated in vitro using knockdown and overexpression cell lines. The effect of PARVA in cell proliferation, migration, and invasion in PCa cells was detected by MTS assay and Transwell assay. Real-time polymerase chain reaction (PCR) and Western blot analysis were used to assess the gene expression in mRNA and protein level.ResultsThe microarray data analysis indicated that PARVA was drastically downregulated in primary and metastatic PCa compared with normal and primary samples, respectively (all P<0.001). Multivariate Cox regression analysis suggested that downregulation of PARVA in PCa was an independent prognostic factor for poor biochemical recurrence (BCR)-free survival (P<0.01). IHC analysis confirmed that PARVA was frequently downregulated in metastatic and primary PCa tissues (All P<0.001). Furthermore, PARVA expression was found to be associated with Gleason score, pathological stage, extracapsular extension, and lymph node invasion (All P<0.05). Knockdown of PARVA triggered cell migration and invasion in vitro, whereas overexpression of PARVA reverted the invasive phenotypes. Mechanistic investigations identified that overexpression of PARVA repressed the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) phosphorylation via inhibiting the integrin-linked kinase (ILK) biological function. With knockdown of ILK, the downregulated MAPK/ERK phosphorylation and Myosin Light Chain 2 (MLC2) expression by PARVA overexpression were abolished, indicating that the PARVA effect on PCa is ILK/MAPK/ERK pathway dependent.ConclusionsOur study revealed that loss of PARVA expression in PCa promotes metastasis by releasing the inhibition of ILK activity, followed by the activation of MAPK/ERK and MLC2 signaling.     

Circ_0061140 stimulates the malignant development of prostate cancer by targeting miR-1193

BackgroundThis study sought to explore the expression pattern in prostate cancer (PCa) tissues, as well as the regulatory effects of circ_0061140 on the proliferative potential of PCa cells.MethodsA quantitative real-time polymerase chain reaction (qRT-PCR) analysis was undertaken to detect circ_0061140 levels in 43 paired PCa tissues and adjacent normal tissues. After the knockdown of circ_0061140, changes in the proliferative potential of PCa cells and tumor growth in nude mice with PCa were detected. Finally, the relationship of circ_0061140 and miR-1193 in the development of PCa was assessed.ResultsThe results showed that circ_0061140 was upregulated in PCa tissues. PCa patients with higher Gleason score or larger sized tumors expressed higher levels of circ_0061140. Additionally, the knockdown of circ_0061140 inhibited the proliferative potential of PCa cells. MiR-1193 was the target gene binding circ_0061140, and its level was negatively regulated by circ_0061140. Finally, rescue experiments showed that miR-1193 was regulated by circ_0061140 in the development of PCa.ConclusionsCirc_0061140 is upregulated in PCa tissues, and is closely linked to Gleason score and tumor size in PCa. Additionally, it causes PCa cells to proliferate by negatively regulating miR-1193.     

Hydropenia may accelerates the progression of orthotopic bladder cancer induced by N-methyl-N-nitrosourea by increasing the expression levels of AQP1, AQP3, and AQP4

BackgroundIncreasing evidence has demonstrated aquaporins (AQPs) to be critical players in carcinogenesis. Here, we aimed to explore the role of hydropenia in the progression of bladder cancer (BCa), as well as to assess the expression of AQP1, AQP3, and AQP4 in bladder tissues from hydropenic and N-methyl-N-nitrosourea (MNU)-treated rats.MethodsAn orthotopic BCa model was induced by administering Sprague Dawley rats with MNU. A hydropenic rat model was established by administrating rats with 2/3 of the amount of water given to the control group. At week 8, the rats were sacrificed and their bladder tissues were collected. Then, pathological alterations in the rat bladders were assessed by hematoxylin and eosin staining. The RNA and protein expression levels of AQP1, AQP3, and AQP4 were determined by using qRT-PCR and western blot assays.ResultsAll of the rats (100%) administrated with MNU developed tumors, of which 5 were large (diameter, 0.5–1.0 cm), 10 were medium (diameter, 0.2–0.5 cm), and 5 were small (diameter, <0.2 cm) in size. The tumors were nodular and cauliflower shaped, with multiple satellite focus, and were accompanied by bleeding, ulcers, stones, and residual urine. Hematoxylin and eosin staining revealed that the bladder mucosa was incomplete, with a large amount of necrotic tissue and obvious leukocytic infiltration. The tumor volume in the MNU + hydropenia group was significantly larger than that in the MNU group. Noticeably, hydropenia exacerbated pathological changes induced by MNU administration. QRT-PCR and western blot analysis revealed that the MNU group, hydropenia group, and MNU + hydropenia group had significantly increased levels of AQP1, AQP3, and AQP4 compared to the control group, with the most dramatic increase seen in the MNU + hydropenia group.ConclusionsHydropenia exacerbates pathological alterations induced by MNU in rats with orthotopic BCa by increasing the expression levels of AQP1, AQP3, and AQP4. This study reveals a possible mechanism of the occurrence of BCa.     

阿司匹林通过抑制STAT3磷酸化下调MCL-1和VEGF mRNA和蛋白表达促进胆囊癌细胞凋亡、抑制增殖

目的 探讨阿司匹林对人胆囊癌细胞株GBC-SD细胞凋亡及增殖的影响及其潜在机制.方法 采用MTT法检测不同浓度梯度及作用时间下阿司匹林对GBC-SD细胞增殖的影响,流式细胞术检测阿司匹林对细胞凋亡的影响;Real-time PCR检测GBC-SD细胞中信号转导和转录激活因子3(STAT3)、髓细胞白血病因子-1(MCL...     

Measuring dynamics of caspase-3 activity in living cells using FRET technique during apoptosis induced by high fluence low-power laser irradiation

BACKGROUND AND OBJECTIVES: Low-power laser irradiation (LPLI) has been used for therapies such as curing spinal cord injury, healing wound etc. Yet, the mechanism of LPLI remains unclear. In order to determine the effects of high fluence LPLI on cell growth and caspase-3 activity, we have measured the dynamics of caspase-3 activity during cell apoptosis induced by high fluence LPLI treatment. STUDY DESIGN/MATERIALS AND METHODS: He-Ne laser was used to irradiate human lung adenocarcinoma cells (ASTC-a-1). Cell Counting Kit-8 was used for cytotoxicity assay. A fluorescent microscope was used to perform fluorescence resonance energy transfer (FRET) imaging. A luminescence spectrometer was used to acquire the fluorescent emission spectrum. Statistical analysis was performed with Student's paired t-test. RESULTS: Cytotoxicity assay showed that when light irradiation fluence exceeded 60 J/cm2, LPLI treatment induced ASTC-a-1 cell apoptosis in a fluence-dependent manner. FRET imaging and spectrofluorometric analysis demonstrated that caspase-3 was activated during high fluence LPLI-induced cell apoptosis. CONCLUSIONS: Using FRET technique, we have reported that high fluence LPLI can induce human lung adenocarcinoma cells (ASTC-a-1) apoptosis. The activation of caspase-3 plays an important role in the apoptotic process.     

Influence of the treatment of developmental dysplasia of the hip by the abduction brace on locomotor development in children

Purpose  

To assess the influence of treating developmental dysplasia of the hip (DDH) with the abduction brace on locomotor development in children.     

人脐带间充质干细胞对耐亚胺培南铜绿假单胞菌耐药性形成的影响

目的探讨人脐带间充质干细胞（hUCMSCs）体外对耐亚胺培南铜绿假单胞菌（IRPA）耐药性形成及对OprD2基因的影响。 方法本实验设立3个组,实验组为hUCMSCs组,对照组为细胞对照组（即人肺成纤维细胞组,NHLF组）和空白对照组。次抑菌浓度肉汤诱导PA耐药传导过程中,hUCMSCs组和NHLF组分别加入其与PA共培育所得的上清液,空白对照组加入细胞培养液,观察3组诱导耐药所需代数以及抑菌圈的大小。诱导耐药前后分别采用K-B法及real-time PCR法测定PA对常见抗菌药物的敏感性及OprD2基因的表达量。 结果经次抑菌浓度的亚胺培南诱导后,hUCMSCs组PA耐药性的出现较NHLF组及空白对照组延迟。NHLF组和空白对照组PA于诱导的第17代出现亚胺培南耐药,而hUCMSCs组PA于第19代出现耐药性。real-time PCR结果显示,诱导耐药后PA中OprD2表达量较诱导前出现减少或消失。其中hUCMSCs组PA OprD2的表达量减少至诱导耐药前的10.96%,而NHLF及空白对照组OprD2无表达,即诱导后出现OprD2基因缺失。 结论人脐带间充质干细胞具有延迟PA耐药性形成的作用,其机制可能是通过分泌抗菌肽LL-37和人β防御素-2从而抑制OprD2表达的减少,而外膜蛋白OprD2表达量减少或缺失是引起PA对亚胺培南耐药的原因。