Toxicity evaluation of Dolichandrone serrulata flower extract on vital and reproductive organs in adult male rats

Abstract Although Dolichandrone serrulata flower (DSF) aqueous extract has been shown to possess pharmacological properties, its systemic toxicity has still to be evaluated. The present study aimed to investigate the sub-chronic toxicity effect of DSF extract on biochemical parameters and histological structures of liver, kidney, testis, and epididymis plus vas deferens. Adult male rats were administered DSF at 100, 300, and 600 mg/kgBW via oral gavage for 48 consecutive days while control rats received distilled water. At the end of the experiment, blood, liver, kidney, testis, and epididymis plus vas deferens samples were collected to determine any changes to serum biochemical components including ALT, ALP, and creatinine levels and histological structures. The results revealed no significant difference in body weight and food or water consumption between control and the DSF-treated groups. It was found that DSF significantly increases the weight of epididymis plus vas deferens, while the kidney and liver showed a decrease in the high dose group (P value 0.05). Histological changes in these vital and reproductive tissues including fibrosis were not observed after administration but ALT, ALP, and creatinine levels were significantly altered in the treated groups (P value 0.05). These altered levels, however, were still within normal ranges. In conclusion, these findings demonstrated that D. serrulata flower extract had no sub-chronic toxicity on vital and reproductive structures but slightly altered some liver and kidney functions.

Resumo Como o extrato aquoso da flor de Dolichandrone serrulata (DSF) demonstrou ter algumas propriedades farmacológicas, sua toxicidade sistêmica ainda não foi avaliada. Este estudo teve como objetivo investigar o efeito da toxicidade subcrônica do extrato de DSF em parâmetros bioquímicos e estruturas histológicas do fígado, rim, testículo e epidídimo mais os vasos deferentes. Ratos machos adultos foram administrados com DSF em 100, 300 e 600 mg / kg de peso corporal por meio de gavagem oral por 48 dias consecutivos, enquanto os ratos controle receberam água destilada. No final do experimento, amostras de sangue, fígado, rim, testículo e epidídimo mais canais deferentes foram coletados para determinar as alterações dos componentes bioquímicos séricos, incluindo ALT, ALP e níveis de creatinina e estruturas histológicas. Os resultados revelaram que o peso corporal e o consumo de comida ou água do controle e de todos os grupos tratados com DSF não foram significativamente diferentes. Verificou-se que o DSF aumentou significativamente o peso do epidídimo mais os canais deferentes, mas o rim e o fígado diminuíram no grupo de alta dose (P valor 0,05). As alterações histológicas, incluindo fibrose de tais tecidos vitais e reprodutivos, não foram encontradas após a administração, mas os níveis de ALT, ALP e creatinina foram significativamente alterados nos grupos tratados (valor P 0,05). No entanto, seus níveis alterados ainda estavam em intervalos normais. Em conclusão, esses achados demonstraram que o extrato da flor de D. serrulata não apresentou toxicidade subcrônica nas estruturas vitais e reprodutivas, mas alterou ligeiramente algumas funções hepáticas e renais.

Estadios de Tanner y orquidometría de Prader en varones adolescentes. Estudio descriptivo transversal / Tanner stages and Prader orchidometry in male adolescents. A descriptive, cross-sectional study

Introducción. La escala de Tanner y el orquidómetro de Prader son los instrumentos más utilizados para evaluar el desarrollo puberal en los niños. La evaluación de la pubertad en la clínica solo es útil si se dispone de datos de referencia recientes y confiables de la misma población para comparar. Objetivo: evaluar la correlación entre los estadios de Tanner y el volumen testicular (VT) en adolescentes argentinos. Población y métodos. Diseño descriptivo transversal, realizado con varones saludables de 9 a 20 años de edad. Se excluyeron varones con patología urogenital y enfermedades que afectan el crecimiento testicular. La correlación entre estadios de Tanner y VT fue evaluada con pruebas no paramétricas. Resultados. Se evaluaron 367 varones con una edad de 13,8 ± 2,5 años. El VT aumentó en correlación a los estadios de Tanner (Spearman 0,943; p <0,001) con volúmenes significativamente diferentes, salvo en los estadios iniciales genital 1-2 (p 0,343) y vello púbico 1-2 (p 0,447). El 16 % (intervalo de confianza del 95 % 9,6-24,4 %; n = 17/106) de los varones peripuberales fue clasificado erróneamente basado en los estadios de Tanner. Conclusiones. Durante la pubertad masculina, el VT aumentó en correlación con los estadios de Tanner, pero no presentó diferencias significativas entre los estadios 1 y 2 de Tanner. Es fundamental el uso del orquidómetro de Prader para detectar el inicio puberal en varones.

Introduction. The Tanner scale and the Prader orchidometer are the instruments most commonly used to assess pubertal development in children. The assessment of puberty in the clinic is only useful if recent and reliable references in the same population are available for comparison. Objective: to assess the correlation between Tanner stages and testicular volume (TV) in Argentine adolescents. Population and methods. Study with a descriptive, cross-sectional design conducted in healthy boys aged 9­20 years. Male children and adolescents with urogenital conditions and disorders affecting testicular growth were excluded. The correlation between Tanner stages and TV was assessed using non-parametric tests. Results. A total of 367 male adolescents with an average age of 13.8 ± 2.5 years were assessed. TV increased in correlation to Tanner stages (Spearman: 0.943, p < 0.001) with significantly different volumes, except in the early genital 1-2 stages (p 0.343) and pubic hair 1-2 stages (p 0.447). Among peripubertal boys, 16% (95% confidence interval: 9.6­24.4%, n = 17/106) were wrongly classified based on Tanner stages. Conclusions. During male puberty, TV increased in correlation to Tanner stages, but no significant differences were observed between Tanner stages 1 and 2. Using the Prader orchidometer is critical to establish the onset of puberty in boys

The effectiveness of insulin therapy in the functional and morphological recovery of leydig cells in rats submitted to Diabetes mellitus

The effects of systemic insulin administration at different concentrations on the testicular tissue of diabetic adult rats, induced by streptozotocin, are evaluated by the morphological analysis of spermatogenic process. Twenty-four adult male rats were divided into 1) Control Group: they received citrate buffer, by intraperitoneal injection; 2) Diabetic Group: induced by intraperitoneal injection of streptozotocin (60 mg. kg-1 of body weight); 3) Insulin 50%: induced diabetes treated with half of standard dosage of insulin; 4) Insulin 100%: induced diabetes treated with standard dose of insulin. After eight weeks, animals were weighted and anesthetized; testicles were removed and processed in resin. Body and testicular weight of diabetic rats decreased when compared to that of control. Parameters increased with insulin therapy. Testosterone levels were low in diabetic animals but rates recovered after insulin therapy. Nuclear diameter and volume of Leydig cells decreased in diabetic rats although they significantly increased after insulin therapy. Results showed that the administration of insulin in diabetic rats promoted a protective effect of testicular parenchyma, enhancing efficient recovery on testosterone levels and increase in daily sperm production.

The probable radioprotective role of Acacia nilotica L. against biochemical and cytogenetic disorders induced in gamma irradiated male rats

Gum Arabic (Acacia nilotica L.) is a respected plant that has many nutrients and curative practices. It hinders, improves, or manages many disorders. The radio-protective activity of Acacia nilotica was investigated against γ-rays-induced testicle damage in rats. Twenty-four rats were correspondingly distributed into 4 groups; control, Acacia nilotica (15mg/kg, daily for 30 days), γ-irradiated (5Gy γ-rays, single dose) and Acacia nilotica plus γ-rays treated groups. The plasma testosterone and total antioxidant status (TAS) were estimated. Lipid peroxidation; malondialdehyde (MDA), reduced glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), also glutathione peroxidase (GPx), catalase (CAT), tumor necrosis factor-α (TNF-α) with interleukin-1ß (IL-1ß), were determined in the testicle tissues. A testis weight, sperm count and motility, peripheral-blood and bone-marrow micronuclei (PMN and BMN), and frequency of chromosomal aberrations (CAs) were scored. A significant decline in the levels of plasma testosterone with TAS observed in the γ-irradiated rats. The results also showed significantly increased levels of testicle MDA, inflammatory markers, PMN, BMN and CAs frequencies and decrease in testes weight, sperm count and motility and levels of testicle antioxidants markers in gamma irradiated group. All these biochemical and fertility indices results were significantly enhanced in the Acacia nilotica plus γ-rays treated groups. However, the possible alleviate activity of Acacia nilotica on γ-rays-induced testicle injury in rats has not previously conversed, and this is the topic of this study.

Histopathological examination of the protective effect of intense exercise in apoptotic germ cell damage due to diabetes

Purpose: The aim of this study was to determine the protective and antioxidative effects of intensive exercise on streptozotocin (STZ)-induced testicular damage, apoptotic spermatognial cells death, and oxidative stress. Methods: 36 male Sprague Dawley rats were divided into three groups: control, diabetes, and diabetes+intensive exercise (IE) groups. Testicular tissues were examined histopathologically and antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) activity, as well as serum testosterone level, were measured. Results: Seminiferous tubules and germ cells were found to be better in the testis tissue of the intense exercise group than in the diabetes group. Diabetes suppressed antioxidant enzymes CAT, SOD, GPx and testosterone levels were significantly decreased, and increased MDA level in the diabetic group compared to diabetes+IE group (p < 0.001). Following four weeks of treatment, intensive exercise improved the antioxidant defense, significantly decreased MDA activity, and increased testosterone levels in testicular tissue in the diabetic group compared to diabetes+IE group (p < 0.01). Conclusion: STZ-induced diabetes causes damage to the testis tissue. In order to prevent these damages, exercise practice has become very popular nowadays. In present study, our intensive exercise protocol, histological, and biochemical analysis of the effect of diabetes on the testicular tissues is shown.

Testicular decompression and tunica vaginalis flap in human acute testicular torsion: modified step-by-step technique description and preliminary outcomes

ABSTRACT Objective To report the effects of a tunica vagina flap on testicular compartment syndrome. Methods This single-arm clinical trial was conducted from September 2020 to October 2021. Consecutive patients with suspected testicular torsion within 24 hours of pain onset were included. Patients with past testicular torsion, bilateral torsion, or previous atrophy were excluded. The tunica vaginalis was opened, and the intratesticular pressure was measured before testicular retrieval from the scrotum and detorsion (P1), after detorsion (P2), and after transverse incision (P3). A tunica vaginalis flap was performed and a new intratesticular pressure was recorded (P4). The contralateral testicular pressure was recorded before fixation (Pc). The minimum follow-up period was 6 months. Results Fifteen patients were recruited from September 2020 to October 2021. Nine patients completed the follow-up. The median age (range) was 15 years (9-19). The mean P1, P2, P3, P4, and Pc (range) were 43, 60, 23, 20, and 14mmHg, respectively. The atrophy rate was 66.3% and the viability was 88.9%. No major complications were observed. Conclusion The modified tunica vaginalis flap in acute testicular torsion decreased intratesticular pressure. Furthermore, normal testicular pressure can improve testicular preservation. It can also decrease testicular pressure to normal levels and preserve the testicular parenchyma.

Ubiquitin protein E3 ligase ASB9 suppresses proliferation and promotes apoptosis in human spermatogonial stem cell line by inducing HIF1AN degradation

Background Spermatogonial stem cells (SSCs) are critical for sustaining spermatogenesis. Even though several regulators of SSC have been identified in rodents, the regulatory mechanism of SSC in humans has yet to be discovered. Methods To explore the regulatory mechanisms of human SSCs, we analyzed publicly available human testicular single-cell sequencing data and found that Ankyrin repeat and SOCS box protein 9 (ASB9) is highly expressed in SSCs. We examined the expression localization of ASB9 using immunohistochemistry and overexpressed ASB9 in human SSC lines to explore its role in SSC proliferation and apoptosis. Meanwhile, we used immunoprecipitation to find the target protein of ASB9 and verified its functions. In addition, we examined the changes in the distribution of ASB9 in non-obstructive azoospermia (NOA) patients using Western blot and immunofluorescence. Results The results of uniform manifold approximation and projection (UMAP) clustering and pseudotime analysis showed that ASB9 was highly expressed in SSCs, and its expression gradually increased during development. The immunohistochemical and dual-color immunofluorescence results displayed that ASB9 was mainly expressed in nonproliferating SSCs. Overexpression of ASB9 in the SSC line revealed significant inhibition of cell proliferation and increased apoptosis. We predicted the target proteins of ASB9 and verified that hypoxia-inducible factor 1-alpha inhibitor (HIF1AN), but not creatine kinase B-type (CKB), has a direct interaction with ASB9 in human SSC line using protein immunoprecipitation experiments. Subsequently, we re-expressed HIF1AN in ASB9 overexpressing cells and found that HIF1AN reversed the proliferative and apoptotic changes induced by ASB9 overexpression. In addition, we found that ABS9 was significantly downregulated in some NOA patients, implying a correlation between ASB9 dysregulation and impaired spermatogenesis. Conclusion ASB9 is predominantly expressed in human SSCs, it affects the proliferation and apoptotic process of the SSC line through HIF1AN, and its abnormal expression may be associated with NOA.

Spermatic cord torsion: a retrospective analysis

ABSTRACT Objective To evaluate the time interval and possible delay in transportation to referral units for the treatment of testicular torsion. Methods We retrospectively analyzed all cases of spermatic cord torsion surgically treated at a university hospital between January 2018 to December 2021. We evaluated the time intervals, including pain onset until the first presentation (D1), interhospital transference time (D2), pain onset until urological evaluation in a tertiary service (D3), urological evaluation until surgery (D4), and time from pain onset to surgical treatment (D5). We analyzed demographic and surgical data, orchiectomy rates, and time intervals (D1-D5). Torsions presented to the first medical presentation within 6h were considered early for testicular preservation. Results Of the 116 medical records evaluated, 87 had complete data for the time interval analysis (D1 to D5) and were considered the total sample. Thirty-three had D1 ≤6h, 53 had D1 ≤24h (includes patients in the D1 ≤6h subgroup), and 34 had D1 >24h. The median time intervals of the total samples and subgroups D1 ≤6h, D1 ≤24h, and D1 >24h were D1 = 16h 42min, 2h 43min, 4h 14min and 72h, D2 = 4h 41min, 3h 39min, 3h 44min and 9h 59min; D3 = 24h, 6h 40min, 7h and 96h; D4 = 2h 20min, 1h 43min, 1h 52min and 3h 44min; D5 = 24h 42min, 8h 03min, 9h 26min and 99h 10min, respectively. Orchiectomy rates of the total sample, subgroups D1 ≤6h, D1 ≤24h, and D1 >24h were 56.32%, 24.24% (p<0.01), 32.08% (p<0.01), and 91.18% (p<0.01), respectively. Conclusion Late arrival at the emergency department or a long interhospital transference time determined a large number of patients who underwent orchiectomy. Thus, public health measures and preventive strategies can be developed based on the data from this study aiming to reduce this avoidable outcome.

Effects of Rooibos (Aspalathus linearis) on the Testicular Parenchyma of Adult BALB/C Mice Submitted to a Hypercaloric Diet

Oxidative stress due to obesity plays a detrimental role in the testicular microenvironment and sperm parameters. We explored the impact of a hypercaloric diet in male BALB/c mice as a condition to trigger damage to the spermatogenic process and the antioxidant effect of Aspalathus linearis as well. We used a hypercaloric diet in animals divided into 3 groups: Control, Hypercaloric diet control (HC) and Hypercaloric diet and Rooibos infusion (HCR). Morphometric parameters, enzyme dosages, cell viability, and tubular histopathology were evaluated. Body weight increased in HCR animals at weeks 3, 4, and 8. We found a reduction in seminiferous epithelium height, with an increase in the tubular diameter of the HCR group. Catalase levels were lower in HC and HCR, while carbonyl protein was decreased in HC. We estimate that it induces oxidative stress (OS) capable of affecting the seminiferous epithelium and that the infusion of A. linearis does not demonstrate a potential benefit in cell preservation.

Histo-Functional Improvement of Mammalian Testis after Platelets-Rich Plasma Treatment

Background: Among new therapies emerging in the medical field, the use of platelet-rich plasma (PRP) in human reproduction has not yet been explored. Platelet-rich plasma (PRP) has a potential effect on tissue repair through proliferation and differentiation of tissue progenitor cells. The aim of this study was to evaluate the effect of PRP on the testis structure and function in the rabbit model. Material and methods: A total of 30 male rabbits were recruited in this study. They were allocated into two groups (15 in each group) to receive an injection of PRP (PRP Group), or normal saline (Control Group) Results: there were statistically significant differences in Means of germinal layer width, Leydig cell number, and Sertoli cell number was significantly higher in the PRP group compared to that in the control group ( P < 0.05). The PRP group had a higher means of sperm concentration and normal morphology compared with the control groups (P < 0.05). Conclusion: the platelet-rich plasma is found to have a good potential effect on the testicular tissue that improved the histological and functional aspects and could be considered a promising future treatment for hypogonadism status in many disorders.

Significance of cancer testis?associated antigens (SPAG9 and FBXO39) in colon cancer

Background: Cancer testis antigens (CTA) are normally expressed in immune privileged tissues such as the testis. They are considered tumor-associated antigens because they are specifically expressed in different cancers. Their distinct nature rendered them appealing targets for cancer diagnosis, prognosis. and immunotherapy. We aimed to identify the association of two CTA genes with colon cancer (CC) in a cohort of Egyptian patients. Methods: We measured the relative gene expression levels of two CTAs: SPAG9 and FBXO39 in colonic tumor tissue and adjacent normal-appearing mucosa in 50 newly diagnosed colon cancer patients by real-time reverse transcription polymerase chain reaction. Gene expression was also studied in relation to demographic and pathological criteria. Results: SPAG9 and FBXO39 were overexpressed in 22% and 40% of cases, respectively. Overexpression of both genes was evident in 14% of cases. We report the significant expression of FBXO39 (P < 0.01) in tumor tissue compared to normal tissue. SPAG9 was significantly increased in large sized tumors compared to smaller sized tumors. Otherwise, there was no significant association between gene expression and the evaluated clinicopathological features (P > 0.05). Conclusions: SPAG9 and FBXO39 are possible CC diagnostic biomarkers. Further studies are warranted to validate our findings.

Expression of the homeobox proteins Dlx-5 and TLX1 during the development of cat testis / Expresión de las proteínas homeobox Dlx-5 y TLX1 durante el desarrollo del testículo de gato

SUMMARY: Recent studies have shown that homeobox proteins play an important role in the formation and development of tissues and organs in the embryonic period. In our study, the distribution of Dlx-5 and TLX proteins, which are members of the homeobox family, in the testis, epididymis and ductus deferens ducts of some cat breeds were investigated. For this purpose, in the study, 18 testes younger than six months (immature) and older than one year (mature) were examined under a light microscope using an immunohistochemical method (indirect streptavidin-biotin complex). While it was determined that Dlx-5 and TLX1 proteins were expressed at varying levels in cells in immature and mature cat testicles, epithelial cells of ductus epididymis and ductus deferens, and smooth muscle cells of ductus deferens, no differences were observed between cat breeds. Dlx-5 immunoreactivity was more intense in the testes, epididymis and deferens ducts of immature and mature compared to TLX1. These results suggested that both proteins play important roles in the development of male feline genital organs and in the secretion and differentiation of cells, and also further observation of Dlx-5 expression suggested that this protein may be more effective than TLX1 in testicular development and physiological processes.

RESUMEN: Estudios recientes han demostrado que las proteínas homeobox juegan un papel importante en la formación y desarrollo de tejidos y órganos en el período embrionario. En nuestro estudio, se investigó la distribución de las proteínas Dlx-5 y TLX, que son miembros de la familia homeobox, en los testículos, en el epidídimo y en los conductos deferentes de algunas razas de gatos. En el estudio fueron examinados, 18 testículos de animales menores de seis meses (inmaduros) y mayores de un año (maduros) bajo un microscopio óptico utilizando un método inmunohistoquímico (complejo indirecto de estreptavidina-biotina). Si bien se determinó que las proteínas Dlx-5 y TLX1 se expresaron en niveles variables en las células de los testículos de gatos inmaduros y maduros, las células epiteliales del epidídimo y del conducto deferente y las células del músculo liso del conducto deferente, no se observaron diferencias entre las razas de gatos. La inmunorreactividad de Dlx-5 fue más intensa en los testículos, epidídimo y conductos deferentes de gatos inmaduros y maduros en comparación con TLX1. Estos resultados sugieren que ambas proteínas tienen un rol importante en el desarrollo de los órganos genitales felinos masculinos y en la secreción y diferenciación de células, y también la observación de la expresión de Dlx-5 sugirió que esta proteína puede ser más efectiva que TLX1 en el desarrollo testicular y en los procesos fisiológicos.

Gene Expression Endpoints Following Subchronic Thiamethoxam Exposure in Adult Male Rabbits

Aim: to investigate the sub chronic toxicity of thiamethoxam on some parameters of reproductive performance in adult male rabbits including gene expression of LDH-C4, FSH? and LH? and GnRHR. Method: sixteen adult male Chinchilla rabbits were divided into two equal groups. Animals in the first group were treated orally with TMX at dose of 250 mg/kg body weight for 90 days. The second group was served as control. Result: Obtained results showed that TMX increased the relative weight of some reproductive organs including testis and prostate. Hormonal analysis revealed that, TMX induced a significant elevation in the serum testosterone level, while the concentrations of FSH and LH hormones did not exhibit any alterations between treated and control groups. In addition, LDH-C4, FSH? and LH? and GnRHR genes were down regulated in TMX treated group. Conclusion: Administration of thiamethoxam for 90 days in male rabbits induced a noticeable adverse effect on serum testosterone level and down regulated genes related to male rabbit reproductive performance

Testicular arteries anatomy applied to fowler-sthephens surgery in high undescended testis - a narrative review

ABSTRACT Objectives: In this review we will describe the testicular vessels anatomy and the implications of these vessels in surgical treatment of high undescended testis. Material and Methods: We performed a narrative review of the literature about the role of the testicular arteries anatomy in the treatment of high undescended testis. We also studied two human testes to illustrate the testicular vascularization. Results: Each testis is irrigated by three arteries: testicular artery (internal spermatic artery), a branch of the right aorta; deferential artery (vasal artery), a branch of the inferior vesicle artery that originates from the anterior trunk of internal iliac artery and cremasteric artery (external spermatic artery), a branch of the inferior epigastric artery. There are important communications among the three arteries with visible anastomotic channels between the testicular and deferential arteries. Conclusions: Laparoscopic transection of the testicular vessels by dividing the spermatic vessels (Fowler-Stephens surgery) is safe in patients with high abdominal testis due to the great collateral vascular supply between testicular, vasal and cremasteric arteries; also, two-stage Fowler-Stephens orchiopexy appears to carry a higher rate of success than the single stage approach.

CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice / 亚洲男科学杂志(英文版)

Gene expression analyses suggest that more than 1000-2000 genes are expressed predominantly in mouse and human testes. Although functional analyses of hundreds of these genes have been performed, there are still many testis-enriched genes whose functions remain unexplored. Analyzing gene function using knockout (KO) mice is a powerful tool to discern if the gene of interest is essential for sperm formation, function, and male fertility in vivo. In this study, we generated KO mice for 12 testis-enriched genes, 1700057G04Rik, 4921539E11Rik, 4930558C23Rik, Cby2, Ldhal6b, Rasef, Slc25a2, Slc25a41, Smim8, Smim9, Tmem210, and Tomm20l, using the clustered regularly interspaced short palindromic repeats /CRISPR-associated protein 9 (CRISPR/Cas9) system. We designed two gRNAs for each gene to excise almost all the protein-coding regions to ensure that the deletions in these genes result in a null mutation. Mating tests of KO mice reveal that these 12 genes are not essential for male fertility, at least when individually ablated, and not together with other potentially compensatory paralogous genes. Our results could prevent other laboratories from expending duplicative effort generating KO mice, for which no apparent phenotype exists.

Flurochloridone-induced apoptosis via IRE1α-JNK signaling pathway in mice testicular cells and TM4 cells / 环境与职业医学

Background Flurochloridone (FLC) can induce apoptosis in Sertoli cells, but the specific mechanism remains unknown. Objective To investigate the testicular cell apoptosis in mice as well as apoptosis and activation of endoplasmic reticulum stress in TM4 cell line induced by FLC through in vivo and in vitro study designs respectively, and study the role of inosital-requiring enzyme 1α (IRE1α)-c-Jun N-terminal kinase (JNK) signaling pathway in the process of FLC-induced apoptosis in TM4 cells through intervention study design. Methods Testicular tissues were collected from male C57BL/6 mice which were treated with 3, 15, 75, and 375 mg·（kg·d）−1 FLC by oral perfusion for 28 d. Apoptosis was observed by TUNEL staining, and the levels of apoptosis-related proteins were detected by Western blotting, including B-cell lymphoma-2 (Bcl-2), Bcl-2 interacting mediator of cell death (Bim), and Bcl-2 associated X protein (Bax). In the in vitro study, TM4 cells were treated with different concentrations of FLC (40, 80, and 160 μmol·L−1) for 6 h, then apoptosis rate was detected by flow cytometry, and the levels of apoptosis-related proteins (Bcl-2, Bim, and Bax) and endoplasmic reticulum stress-related proteins [glucose regulated protein 78 (GRP78), phosphorylated-protein kinase R like endoplasmic reticulum kinase (p-PERK), activating transcription factor 6 (ATF6), phosphorylated-inosital-requiring enzyme 1α (p-IRE1α), and phosphorylated-JNK (p-JNK)] were measured by Western blotting. In the intervention study, TM4 cells were pretreated with IRE1α phosphorylation inhibitor 4μ8C and JNK phosphorylation inhibitor SP600125 for 6 h, then treated with 160 μmol·L−1 FLC for 6 h. The levels of apoptosis-related proteins and endoplasmic reticulum stress-related proteins were measured by Western blotting, and cell viability was detected by cell counting kit-8. Results After the male C57BL/6 mice orally exposed to FLC for 28 d, apoptosis occurred in the seminiferous tubule. The protein expression level of Bcl-2, apoptosis inhibitor, was decreased in the 75 and 375 mg·(kg·d)−1 groups (P<0.05), and the protein expression levels of Bim and Bax, apoptosis promoters, were increased in the 75 and 375 mg·(kg·d)−1 groups respectively (P<0.05). The percentages of apoptotic cells in the 0, 40, 80, and 160 μmol·L−1 FLC groups were 2.7%±0.2%, 4.8%±1.3%, 9.4%±0.3%, and 13.2%±0.2%, respectively, increased significantly compared with the control group (P<0.05). The protein expression level of Bcl-2 also was decreased in the 160 μmol·L−1 FLC group (P<0.05), while the levels of Bim and Bax were increased in both of the 80 and 160 μmol·L−1 groups (P<0.05). The expression levels of endoplasmic reticulum stress-related proteins (GRP78, p-PERK, ATF6, p-IRE1α, and p-JNK) were increased (P<0.05) or showed a rising trend in TM4 cells. Pre-treatment with 4μ8C (25 and 50 μmol·L−1) and SP600125 (10 and 20 μmol·L−1) significantly down-regulated the protein expression levels of GRP78, p-IRE1α, p-JNK, and Bax induced by FLC (P<0.05) or in a downward trend. Both of the inhibitors alleviated the decreased cell viability induced by FLC (P<0.05) or in alleviating fashion. Conclusion FLC could induce apoptosis in mice testis and TM4 cell apoptosis through activating endoplasmic reticulum stress and IRE1α-JNK signaling pathway.

Expression of retinoic acid signaling pathway in mouse damaged testes induced by procymidone during adolescence / 环境与职业医学

Background Procymidone (PCM) exposure can cause damage to reproductive organs of male mice, but whether its mechanism is related to the retinoic acid (RA) signaling pathway is unclear. Objective To explore the possible mechanism of PCM-induced testes damage in adolescent mice. Methods Three-week-old ICR mice (n=64) were randomly divided into a control group and three dose groups (low, medium, and high), with 16 mice in each group. PCM was administered orally at 0, 50, 100, and 200 mg·kg−1·d−1 for 21 consecutive days. Serum and bilateral testes in each mouse were collected to detect content of testosterone in serum and to observe histological changes in testis section after the mice were sacrificed one week after cessation of drug administration. Real-time fluorescence quantitative PCR and Western blotting were used to detect the mRNA expression abundances of genes related to the RA signaling pathway and apoptosis genes Casp9 and Casp12, and the protein expression levels of CYP26A1, ALDH2, and CASP9 respectively. Results Compared with the control group, there was no significant change in the overall appearance and testicular appearance of mice in each dose group after the PCM exposure. According to pathological section observation, the testicular seminiferous tubules of mice in the low-dose group showed slight atrophy and reduced sperm production; the testes of mice in the medium- and the high-dose groups showed obvious pathological damage (e.g. dilated lumen of seminiferous tubules, damaged spermatogenic epithelium, decreased number of spermatogonia, and partial absence of sertoli cells); as the concentration of PCM increased, the degree of spermatogenic epithelial damage in mice gradually increased and the number of spermatozoa in the seminiferous tubules decreased. There were no significant differences in the distance between the anus and the genitals, testicular mass, testicular volume, and testicular organ coefficient among the four groups of mice (P>0.05). The body weights of the mice in the low-, medium-, and high-dose groups were (34.91±1.89), (34.88±1.75), and (32.94±1.37) g respectively, and that in the high-dose group was lower than that in the control group, (35.93±1.99) g, (P<0.05); the serum testosterone concentrations were (313.77±5.32), (305.31±3.47), and (304.80±5.28) pg·mL−1 respectively, which were lower than that in the control group, (319.05±1.92) pg·mL−1 (P<0.05); as the dose of PCM increased, the body weight and serum testosterone concentration showed decreasing trends. The mRNA expression levels of Stra6 and Rbp1 in the high-dose group were higher than those in the control group (P<0.05); the mRNA expression levels of Aldh2, Aldh1a1, Aldh1a3, Rarα, Rar\begin{document}$\beta $\end{document}, Rxrα and Rxr\begin{document}$\beta $\end{document} in the medium-and the high-dose groups were higher than those in the control group (P<0.05); the mRNA expression levels of Cyp26a1 and Cyp26b1 in the medium- and high-dose groups were lower than those in the control group(P<0.05); the mRNA expression levels of apoptosis genes Casp9 and Casp12 in the medium-and the high-dose groups were higher than those in the control group (P<0.05). The protein expression level of CYP26A1 in each exposure group was lower than that in the control group (P<0.05), and the expression level decreased with increasing concentration of PCM; the expression level of ALDH2 protein in the medium- and the high-dose groups and the protein expression level of CASP9 in each exposure group were higher than those in the control group (P<0.05), and the levels increased with increasing concentration of PCM. Conclusion PCM can damage the testis tissues of adolescent mice, where RA signaling pathway, Casp9 and Casp12 genes, and CASP9 protein are activated.

Research status of related factors and recent advances in the diagnosis and treatment of testicular atrophy after orchiopexy in children / 中华泌尿外科杂志

Cryptorchidism is one of the common diseases in children’s genitourinary system. Surgery is the first choice for clinical treatment. Testicular atrophy is a serious complication after operation. The related risk factors of testicular atrophy include the age of operation, the position of testis before operation and the mode of operation. At present, the possible treatments for testicular atrophy are hormone therapy, stem cell therapy and so on. This article reviews the related risk factors, fertility, diagnosis and treatment of testicular atrophy after orchiopexy in children.

Effects of thoracic X-ray irradiation on the spermatogenesis of adult male mice / 中华放射医学与防护杂志

Objective:To investigate the effect of thoracic X-ray irradiation on the spermatogenesis of adult male mice.Methods:A total of 24 healthy adult male C57BL/6 mice (6-8 weeks old) were randomly divided into radiation group (Radiation) and sham-radiation group (Sham), 12 mice in each group. The area of thoracic irradiation was 1.5 cm× 2 cm, and the dose rate was 3.04 Gy/min, 8 Gy/d for 3 consecutive days, 24 Gy in total. At 7 d and 21 d after thoracic irradiation, the bilateral testes and epididymal tails were stripped and the testicular index was calculated. The morphology of testis was examined by haematoxylin-eosin (HE) staining, then the diameter of seminiferous tubules and the thickness of seminiferous epithelium were measured. The sperms were collected from the bilateral epididymal tails for sperm counting. The level of apoptosis in testis and levels of apoptosis-related proteins were detected by TUNEL and Western blot, respectively.Results:Compared with Sham group, the morphology of testis and epididymis was seriously damaged, the diameter of seminiferous tubules significantly decreased at 21 d after irradiation ( t = 8.93, P < 0.05), and the seminiferous epithelium significantly decreased at 7 d and 21 d after irradiation ( t = 4.24, 12.77, P < 0.05). In addition, the number of sperms significantly decreased ( t = 4.30, 2.98, P < 0.05). The number of TUNEL positive cells in the seminiferous epithelium significantly increased at 7 d and 21 d after irradiation ( t = -2.73, -3.74, P < 0.05). Meanwhile, the level of cleaved Caspase-3 protein significantly increased at 7 d and 21 d after irradiation ( t = -2.96, -2.46, P < 0.05). The concentrations of SCF and GDNF did not change at 7 d after irradiation, but were significantly increased at 21 d after irradiation ( t = -10.46, -5.42, P < 0.05). Conclusions:The thoracic X-ray irradiation could lead to spermatogenesis disorder in male adult mice, and the induction of spermatogenic cell apoptosis and the secretory dysfunction of sertoli cells may be involved.

Acutely applied melatonin prevents CCl4-induced testicular lesions in rats: the involvement of the oxidative capacity and arginine metabolism

Abstract Carbon tetrachloride (CCl4) represents an organic chemical that causes reactive oxygen species derived organ disturbances including male infertility. Melatonin (MLT) is a neurohormone with strong antioxidant capacity, involved in numerous physiological processes. In this study we evaluated the capability of MLT, administered in a single dose of 50 mg/kg, to preserve the testicular tissue function after an acute administration of CCl4 to rats. The disturbance in testicular tissue and the effects of MLT after CCl4 exposure were estimated using biochemical parameters that enabled us to determine the tissue (anti)oxidant status and the intensity of arginine/nitric oxide metabolism. Also, the serum levels of testosterone and the histopathological analysis of tissue gave us a better insight into the occurring changes. A significant diminution in tissue antioxidant defences, arginase activity and serum testosterone levels, followed by the increased production of nitric oxide and extensive lipid and protein oxidative damage, was observed in the CCl4-treated group. The application of MLT after the CCl4 caused changes, clearly visible at both biochemical and histological level, which could be interpreted mainly as a consequence of general antioxidant system stimulation and a radical scavenger. On the other hand, the application of MLT exerted a limited action on the nitric oxide signalling pathway.