Objective: To investigate the clinicopathological characteristics of testicular biopsies from Klinefelter syndrome (KS) patients. Methods: The testicular biopsy specimens of 87 patients with KS (a total of 107 biopsy specimens) were collected from the Department of Pathology, Peking University Third Hospital, Beijing, China from January 2017 to July 2022. All patients were diagnosed as KS by peripheral blood karyotyping analysis. The testicular histopathologic features, testicular volume and hormone levels were evaluated retrospectively. The histopathologic analysis was used to assess the quantity and morphology of Leydig cells, the spermatogenic state of seminiferous tubules, the thickening of the basement membrane of seminiferous tubules and the changes of stroma. Results: Leydig cell proliferative nodules were seen in 95.3% (102/107) of KS testicular biopsy tissues. The eosinophilic inclusion bodies and lipofuscin in Leydig cells were found in 52.3% (56/107) and 57.9% (62/107) of specimens, respectively. The Sertoli cell only seminiferous tubules and the hyalinized tubules were found in 66.4% (71/107) and 76.6% (82/107) of the examined tissues, respectively. The tubules with complete spermatogenic arrest were found in 15.9% (17/107) of specimens, and 5.6% (6/107) of the specimens showed low spermatogenesis or incomplete spermatogenic arrest. In 85.0% (91/107) of the specimens, increased thick-walled small vessels with hyaline degeneration were identified. Conclusions: The most common features of KS testicular specimens are Leydig cell proliferative nodules, hyaline degeneration of seminiferous tubules and proliferation of thick-walled blood vessels. Testicular biopsy specimens of KS are rare. The pathologists can make a tentative diagnosis of KS based on the histological findings, combined with the ultrasound and laboratory results, which is helpful for further diagnosis and treatment of KS.
Male ; Humans ; Testis/pathology* ; Klinefelter Syndrome/pathology* ; Retrospective Studies ; Seminiferous Tubules/pathology* ; Biopsy

AIMTo evaluate the long term effect of experimental cryptorchidism on germ cell apoptotic rate and testicular sperm content in adult rats.

METHODSBilateral cryptorchidism was created in 40 adult male Sprague-Dawley rats by surgically manipulating the testes into the abdominal cavity and closing the internal inguinal ring. The rats were sacrificed and the testes removed 6 hours and 2, 4, 7, 21, 28 and 56 days after cryptorchidism. Germ cell apoptosis was quantified by means of TUNEL assay and apoptosis was confirmed using transmission electron microscopy.

RESULTSThe rate of apoptosis peaked at 4 days of cryptorchidism and then progressively declined to a nadir at 14 days of cryptorchidism. At 56 days of cryptorchidism, the germinal epithelium was largely depleted by the apoptotic process and only a few mature sperm were seen within the testis. At this point, a few tubules were seen to be repopulating with primary spermatocytes and the level of germ cell apoptosis began to increase marginally. Testicular sperm count (TSC) began to decline rapidly at day 7 of cryptorchidism. Only a few mature sperm were found in the testes of rats following 56 days of cryptorchidism. Multinucleated giant cells (MGC) were most numerous within the seminiferous tubules at day 4. At day 7, 35 % of MGCs were TUNEL positive. At all subsequent time points, however, MGCs fail to stain positive for apoptosis. This resumption of increased apoptosis coincided with the appearance of a population of primary spermatocytes in some seminiferous tubules. Moreover, there was not a corresponding increase in the number of mature sperm after 56 days of cryptorchidism.

CONCLUSIONThe decline in germ cell apoptosis after 4 days of cryptorchidism can be attributed to be the result of an overall depletion of germ cells. It appears that after a prolonged cryptorchidism (56 days), there is a limited resumption of spermatogenesis presumably as a result of a decrease in the maturing germ cells undergoing programmed cell death.

Animals ; Apoptosis ; Cryptorchidism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; pathology ; Sperm Count ; Spermatozoa ; pathology ; Testis ; pathology

The objective of this study was to investigate the effects of chronic stress on the testes of prepubertal and adult rats and to evaluate whether any alterations could be reversed when stress induction is ended. Seventy-six male rats were assigned to eight groups depending on the type of treatment (control or stressed), the age at which stress was initiated (prepubertal or adult), and the time of evaluation (immediate or late). Stress stimuli were applied for 6 weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimulus were included in SP-I and SA-I groups, respectively. The late prepubertal (SP-L) and adult (SA-L) groups of stressed rats were evaluated 6 weeks after the last stress stimulus. Age-matched rats were used as controls (CP-I, CA-I, CP-L, and CA-L groups). Application of stress stimuli to rats in the SP-I group resulted in body weight and seminiferous tubule diameter reduction. The rats in the SA-I group also showed several functional (testosterone level and sperm parameter) and morphological (testicular weight and seminiferous tubule diameter) reductions. The rats in the SP-L group showed increased body weight and intertubular compartment volumetric and absolute densities and reduced tubular compartment volumetric density. The rats in the SA-L group presented only reduced sperm viability. Stress stimuli promoted changes in the rats in all the study groups. The testes of the adult rats were the most affected by chronic stress. However, the stressed adult rats recovered well from the testicular alterations.
Aging/pathology* ; Animals ; Body Weight ; Chronic Disease ; Male ; Organ Size ; Rats ; Rats, Wistar ; Restraint, Physical ; Semen Analysis ; Seminiferous Tubules/pathology* ; Spermatogenesis ; Stress, Psychological/pathology* ; Testis/pathology* ; Testosterone/blood*

It has become increasingly evident within the last few decades that immunologic factors are involved in some aspects of the reproductive process and hence in the physiology and pathology of genital tract. Clear-cut demonstration of the antigenic power of the spermatozoa or of whole semen in heterogeneous inoculation was first presented toward the end of last century. Around 1952, real progress widening the conception of spermatogenesis was made when a selective destruction of germinal cell! was obtained in guinea pig by auto- or homologous sensitization with a single dose of homogenate prepared from testicular tissue to which Freund`s complete adjuvant was added. The testis lesions were accompanied by development of humoral antibodies and cellular immunity. The purpose of this study is to observe the effect of homologous sensitization with homogenated testicular tissue on the spermatogenesis and immune response in mice, dividing into three groups; the first group is to give a complex of testicular extracts and Freund`s complete adjuvant (10 mice), the second group is to give a Freund's complete adjuvant alone (5 mice), the third group is normal control group (5 mice). The results were as follows: 1. The histopathologic observations revealed that spermatogenesis was more or less adversely affected exception case in group I whereas it unaffected in group II and III. The impairment of spermatogenesis was, diminished number of spermatozoa, degenerated and exfoliated germinal cells in seminiferous tubules and epididymides. 2. Diffusion test and sperm agglutination test for antibody were negative in group I as well as group II and III, which suggested that the histopathologic changes might be caused by cell mediated immune response rather than humoral antibody.
Animals ; Antibodies ; Diffusion ; Fertilization ; Guinea Pigs ; Immunity, Cellular ; Immunologic Factors ; Mice* ; Pathology ; Physiology ; Semen ; Seminiferous Tubules ; Sperm Agglutination ; Spermatogenesis* ; Spermatozoa ; Testis

OBJECTIVETo determine whether testosterone-induced intra-testicular testosterone withdrawal and therefore spermatogenic impairment is associated with looser arrangement of spermatogenic cells in rats.

METHODSAdult male SD rats received intramuscular injection of testosterone undecanoate at 19 mg/(kg x 15 d) for 130 days, and then testicular tissue blocks were obtained for the preparation of methacrylate resin-embedded sections and observation of the changes in testicular histology.

RESULTSApart from such changes as impaired spermiogenesis and spermiation, apparently looser arrangement of spermatogenic cells was seen in 11.5% of the seminiferous tubule profiles, with radial cracks (empty spaces) running towards the tubule lumen being formed between lines, bundles or groups of spermatogenic cells (mainly spermatids and spermatocytes).

CONCLUSIONLooser arrangement of spermatogenic cells is one of the key histological changes resulting from intra-testicular testosterone withdrawal in rats.

Animals ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; cytology ; drug effects ; Spermatogenesis ; drug effects ; Testis ; cytology ; drug effects ; pathology ; Testosterone ; administration & dosage ; adverse effects

AIMTo observe the acute effect of the organophosphorous insecticide malathion on testicular function in mice.

METHODSThe effects of a single dose of malathion [240 mg/kg (1/12 LD(50))] on plasma acetylcholinesterase (ACE) activity, spermatozoa (epididymal cauda counts and teratozoospermia), testis and plasma testosterone concentration) were evaluated at day 1, 8, 16, 35 and 40 after treatment.

RESULTSThe sperm count was decreased significantly 24 h after treatment and teratozoospermia was increased at day 35 and 40. The height of the seminiferous epithelium and the diameter of tubular lumen were decreased at day 8. The percentage of tubular blockade was increased between day 8 and 35. A decrease in testosterone plasma level was observed at day 16 after treatment.

CONCLUSIONMalathion damages male reproduction. The depletion of seminiferous tubules and the increase in teratozoospermia may be a genotoxic damage to the renewing spermatogonia, but the possibility of spermatogenic/spermiogenic disfunction due to a decrease in the plasma testosterone level can not be ruled out.

Animals ; Insecticides ; pharmacology ; Malathion ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Seminiferous Tubules ; drug effects ; pathology ; Sperm Count ; Spermatogenesis ; drug effects ; Spermatozoa ; abnormalities ; Testosterone ; antagonists & inhibitors ; blood ; Time Factors

AIMTo quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion.

METHODSFourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate (EDS, 75 mg/kg) and the same number of animals were injected with normal saline as a control. At days 7 and 12 (after treatment), respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods.

RESULTSThe EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers (per testis) of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively.

CONCLUSIONThe primary spermatogenic lesions following EDS administration were (i) spermiation failure and (ii) detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.

Animals ; Epididymis ; drug effects ; pathology ; Injections, Intraperitoneal ; Leydig Cells ; drug effects ; pathology ; Male ; Mesylates ; administration & dosage ; toxicity ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; pathology ; Testis ; cytology ; drug effects ; growth & development ; pathology

PURPOSE: Testicular microlithiasis (TM) is a relatively rare clinical entity of controversial significance characterized by the existence of hydroxyapatite microliths located in the seminiferous tubules. The aim of this study was to observe the natural course of changes in the calcific density of pediatric TM. MATERIALS AND METHODS: We included a total of 23 TM patients undergoing scrotal ultrasound (US) on at least two occasions from July 1997 to August 2014. We retrospectively analyzed the patient characteristics, clinical manifestations, specific pathological features, and clinical outcomes. We measured the calcified area and compared the calcific density between the initial and final USs. RESULTS: The mean age at diagnosis was 11.3+/-4.6 years, and the follow-up period was 79.1+/-38.8 months (range, 25.4-152.9 months). During the follow-up period, no patients developed testicular cancer. Calcific density on US was increased in the last versus the initial US, but not to a statistically significant degree (3.74%+/-6.0% vs. 3.06%+/-4.38%, respectively, p=0.147). When we defined groups with increased and decreased calcification, we found that diffuse TM was categorized into the increased group to a greater degree than focal TM (10/20 vs. 4/23, respectively, p=0.049). In addition, five of eight cases of cryptorchidism (including two cases of bilateral cryptorchidism) were categorized in the increased calcification group. CONCLUSIONS: Diffuse TM and cryptorchidism tend to increase calcific density. Close observation is therefore recommended for cases of TM combined with cryptorchidism and cases of diffuse TM.
Adolescent ; Calcification, Physiologic ; *Calculi/complications/epidemiology/pathology/physiopathology ; Child ; Cryptorchidism/diagnosis/etiology ; Densitometry/methods ; Follow-Up Studies ; Gonadoblastoma/diagnosis/etiology ; Humans ; Male ; Republic of Korea ; Scrotum/*ultrasonography ; Seminiferous Tubules/*pathology ; *Testicular Diseases/complications/epidemiology/pathology/physiopathology ; *Testicular Neoplasms/diagnosis/epidemiology/etiology

AIMTo evaluate the immunohistopathological changes in the contralateral testis of rats after an experimental spermatic cord torsion.

METHODSMale Sprague-Dawley rats of 45-50 days old were subjected to a 720 degree unilateral spermatic cord torsion for 10, 30 and 80 days (experimental group, E), respectively or sham operation (control group, C). Histopathology of the contralateral testis as well as germ cell apoptosis were studied using the Terminal Deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) technique. The number of testicular lymphocytes, mast cells and macrophages, and the expression of tumor necrosis factor-alpha (TNF-alpha) and its receptor (TNFR1) in testicular cells of the contralateral testis were quantified by histochemistry and immunohistochemistry. TNF-alpha concentration in testicular fluid was determined by ELISA.

RESULTSIn the contralateral testis of rats from the E group, the maximal degree of damage of the germinal epithelium was seen 30 days after torsion. At this time we observed in the E group vs. the C group increases: (i) the number of testicular T-lymphocytes; (ii) the number of testicular mast cells and macrophages; (iii) the percentage of macrophages expressing TNF-alpha; (iv) TNF-a concentration in testicular fluid; (v) the number of apoptotic germ cells; and (vi) the number of TNFR1+ germ cells.

CONCLUSIONExperimental spermatic cord torsion induces, in the contralateral testis, a focal damage of seminiferous tubules characterized by apoptosis and sloughing of germ cells. Results suggest humoral and cellular immune mediated testicular cell damage in which macrophages and mast cells seem to be involved in the induction of germ cell apoptosis through the TNF-alpha/TNFR1 system and in the modulation of the inflammatory process.

Animals ; Apoptosis ; Disease Models, Animal ; Functional Laterality ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; pathology ; Spermatic Cord Torsion ; pathology ; surgery ; Testis ; pathology ; Tumor Necrosis Factor-alpha ; analysis

The toxicity of acrylamide was evaluated through mutagenicity of Salmonella, chromosome aberration of Chinese hamster lung fibroblasts, micronucleus formation in mice and reproductive toxicity in rats. Based on Ames test, acrylamide showed mutagenic potency for strains TA98 and TA100. Moreover, both chromosomal aberration assay and micronucleus assay indicated that acrylamide might have genotoxic potency; the chromosomal aberration frequencies were observed to be proportional to acrylamide concentrations of 5-50 mM, and acrylamide significantly increased micronuclei in peripheral blood cells of mice at doses of higher than 72.5 mg/kg. Male rats were treated with acrylamide at doses of 0, 5, 15, 30, 45, or 60 mg/kg/day for 5 consecutive days, and the toxicity of acrylamide was observed. In the group treated with the highest dose of acrylamide (60 mg/kg/day), the loss of body weight and reduced testis weight were observed. Also the epididymides weights were reduced significantly in all the groups treated with acrylamide. The number of sperms in cauda epididymidis decreased significantly in an acrylamide dose-dependent manner. Rats treated with 60 mg/kg/day of acrylamide showed several histopathological lesions in the seminiferous tubules. There were thickening and multiple layering of the tubular endothelium, and the formation of many multinucleated giant cells in seminiferous tubules. Taken together, acrylamide not only causes the genotoxicity of eukaryotic cells and mice but also shows the toxicological effects on reproductive system in male rats.
Acrylamide/*toxicity ; Animals ; Body Weight ; Carcinogens/*toxicity ; Chromosome Aberrations/chemically induced ; Cricetinae ; Cricetulus ; Epididymis/*drug effects/pathology ; Histocytochemistry ; Male ; Mice ; Mice, Inbred ICR ; Micronucleus Tests ; Mutagenicity Tests ; Organ Size ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules/*drug effects/pathology ; Sperm Count