Secondary Sexual Characteristics Research Articles

Analysis of single nucleotide polymorphisms of the metabotropic glutamate receptors in a transgender population.

Gender incongruence (GI) is characterized by a marked incongruence between an individual's experienced/expressed gender and the assigned sex at birth. It includes strong displeasure about his or her sexual anatomy and secondary sex characteristics. In some people, this condition produces a strong distress with anxiety and depression named gender dysphoria (GD). This condition appears to be associated with genetic, epigenetics, hormonal as well as social factors. Given that L-glutamate is the major excitatory neurotransmitter in the central nervous system, also associated with male sexual behavior as well as depression, we aimed to determine whether metabotropic glutamate receptors are involved in GD. We analyzed 74 single nucleotide polymorphisms located at the metabotropic glutamate receptors (mGluR1, mGluR3, mGluR4, mGluR5, mGluR7 and mGluR8) in 94 transgender versus 94 cisgender people. The allele and genotype frequencies were analyzed by c2 test contrasting male and female cisgender and transgender populations. The strength of the associations was measured by binary logistic regression, estimating the odds ratio (OR) for each genotype. Measurement of linkage disequilibrium, and subsequent measurement of haplotype frequencies were also performed considering three levels of significance: P ≤ 0.05, P ≤ 0.005 and P ≤ 0.0005. Furthermore, false positives were controlled with the Bonferroni correction (P ≤ 0.05/74 = 0.00067). After analysis of allele and genotypic frequencies, we found twenty-five polymorphisms with significant differences at level P ≤ 0.05, five at P ≤ 0.005 and two at P ≤ 0.0005. Furthermore, the only two polymorphisms (rs9838094 and rs1818033) that passed the Bonferroni correction were both related to the metabotropic glutamate receptor 7 (mGluR7) and showed significant differences for multiple patterns of inheritance. Moreover, the haplotype T/G [OR=0.34 (0.19-0.62); P<0.0004] had a lower representation in the transgender population than in the cisgender population, with no evidence of sex cross-interaction. We provide genetic evidence that the mGluR7, and therefore glutamatergic neurotransmission, may be involved in GI and GD.

Markers of Fertility in Adolescents With Chronic Endocrinopathies at Transition From Paediatric to Adult Care

ABSTRACTObjectiveDuring the process of transition from paediatric to adult health care, counselling concerning fertility is an important issue and is based mainly on serum markers of gonadal function. Here, we analysed these markers in adolescents with various underlying endocrine diseases at the time of transition.MethodsAfter reaching near adult height and late puberty (girls: bone age [BA] ≥14 years, and boys: BA ≥16 years), we assessed stages of puberty according to Tanner and measured testes or ovarian volumes and serum markers of gonadal function (anti‐Mullerian hormone [AMH], inhibin B, 17β‐estradiol, testosterone).ResultsOne hundred and ten patients (56 females and 54 males) were included from May 2010 to March 2016 with multiple pituitary hormone deficiency (MPHD; n = 17), growth hormone deficiency (GHD; n = 35), Turner syndrome (TS; n = 27), short stature after being born small for gestational age (SGA; n = 20) and Klinefelter syndrome (KS; n = 11). Female and male adolescents exhibited mature secondary sexual characteristics. The levels of serum inhibin B and AMH were lower in TS and female MPHD than in GHD and SGA, each independently (p &lt; 0.05). The levels of serum AMH were higher whereas serum inhibin B were lower in male MPHD and KS (p &lt; 0.05). Ovary volumes were significantly smaller in patients with TS, and testicular volumes were smaller in patients with KS.ConclusionsAfter current established treatments with sex steroids, the development of secondary sexual characteristics was mature. However, impaired markers of fertility have been identified in patients with TS, KS and MPHD, reflecting gonadal dysgenesis in TS and KS, but gonadal immaturity in MPHD as gonadal gonadotropin stimulation is lacking throughout development. Consequently, in patients with MPHD, these markers cannot reliably predict individual fertility, which warrants consideration and incorporation in future treatment concepts.

Changes in secondary sexual characteristics of female red swamp crayfish (Procambarus clarkii) and relationship to ovarian development: Implications for intensive breeding of seedlings

A rapid selection method of mature Procambarus clarkii females for use in intensive breeding of seedlings is proposed based on evaluation of secondary sexual characteristics and variation patterns in this study. Following regular sampling in ponds, the weight, cephalothorax length, annulus ventralis length, body width, white patch coverage (WC) on the uropod, and uropod content weight-to-weight (WW) ratio of female P. clarkii were found to change significantly during ovarian development. The random forest model identified white patches on the uropod as secondary sexual characteristics of female P. clarkii. In fact, the ovarian maturity status could be determined based on the variable pattern of these white patches (distribution shape, coverage, and density), particularly WC. White patches waxed and waned during ovarian development and oviposition. Specifically, white patches were absent from the uropod of female P. clarkii before the avitellogenic stage; visible on the uropod in the early vitellogenic stage (WC < 30%); WC 30–45% occurred in the midvitellogenic stage; WC > 45% occurred in the late vitellogenic stage; and white patches disappeared from the uropod of female P. clarkii in the post-vitellogenic and resorptive stage. The accuracy of identifying mature female P. clarkii (late vitellogenic stage) based on the white patch pattern was 86.36%. Hence, the observation of white patch changes on the uropod might represent an effective method for rapidly identifying mature female P. clarkii with the potential to improve artificial intensive breeding.

The Role of MRI in Diagnosing Mayer-Rokitansky-Kuster-Hauser Syndrome: A Case Study

The Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome is a rare condition that results from the disturbance of embryonic paramesonephric duct development, which gives rise to varying degrees of malformation of reproductive organs. It is characterized by uterovaginal aplasia with normal secondary sexual characteristics and 46,XX karyotype. We report a 15-year-old female patient with MRKH. Pelvic MRI revealed cervical and uterine agenesis with the absence of the vagina. The diagnosis was confirmed based on radiological findings. The correct clinical and radiological diagnosis of MRKH by MRI is crucial for long-term management.

Proto-Endocrinological Theories of Masculinity/Femininity (1490-1904).

From the mid-seventeenth century, resorption of a testicular "ferment" and resorption of some part of the semen constituted reputable accounts of secondary sexual characteristics. Only in the early twentieth century was the latter, "recrementitious secretion" theory, explicitly considered superseded by one of internal secretion, an advance ushering in the hormone era. A reconstruction of these proto-endocrinological concepts is offered onward from the first, 1490 print edition of Galen's On Semen. Early modern physicians picking up from Galen deliberated widely on the medium and pathway of male and female testicular influences on "the entire body," including the mind, causing "femininity" and "masculinity" in physical, mental-temperamental, and behavioral terms. A switch is discernible from "heat and strength" (Galen) to blood-borne "virility" or testicular vapor (such as proposed in 1564 by Tomás Rodrigues da Veiga), to iatrochemical postulations of a "seminal ferment" (suggested in the late 1650s, perhaps independently, by Thomas Willis at Oxford and Lambert van Velthuysen in Utrecht), finally to a "seminal recrement" or "reabsorbed semen" concept soon after (emergent in the posthumous work of Giovanni Alfonso Borelli, among others). During the late eighteenth century, mounting controversy surrounded both the very idea of that concept and the involved anatomical pathways, informed by multiple experiments.

Infestation of the spider crab Menaethius monoceros (Epialtidae: Brachyura) from the Egyptian Red Sea coasts by Parasacculina pilosella (Polyascidae: Rhizocephala: Cirripedia

The results of this study showed that out of 287 individuals (135 males and 152 females) of the spider crab Menaethius monoceros collected from the Red Sea and the Gulfs of Suez and Aqaba, 15 individuals (6 males and 9 females) were infected with Parasacculina pilosella (Van Kampen & Boschma, 1925) (Polyascidae: Rhizocephala: Cirripedia: Crustacea). A total of 20 externae were recorded, including 11 singles, three doubles, and only one triple on the infected crab abdomens. The overall infection rate was 5.23% for the combined sexes at all sites, declined to 4.44% in males, but increased to 5.92% in females, reaching a higher rate of 16.07% in the populations of Marsa Alam and Shalateen sectors during summer. The examined externae have variable colors, sizes, and shapes based on their sexual maturity, which has been classified into four stages: immature or virgin, early mature, maturing, and late (spent externae). The largest number of externae was eight extruded on the 3rd abdominal segment, while it decreased to six, two, and one on the 4th, 2nd and 5th segments, respectively. The size of externae varied from 1.3 × 1.0 × 1.0 to 6.3 × 4.5 × 2.5 mm (Breadth × Height × Thickness). These results showed changes in the external secondary sexual characters of the infected males, represented by the remarkable broadness of the abdomen fringed with marginal setae and invagination of the sternum, but were limited in infected females.

Pubertal Delay Secondary to Hashimoto's Disease in the Hypothyroid Phase: About a Case in the Internal Medicine Department of the Gabriel Touré University Hospital in Bamako, Mali

Introduction: Pubertal delay is defined by the absence of development of secondary sexual characteristics beyond the age of 13 in girls or 14 years in boys. In children of puberty age, delayed puberty is common in long-standing, untreated hypothyroidism. We report a case of delayed puberty revealed by Hashimoto's disease in the hypothyroid phase diagnosed in the Internal Medicine department of the Gabriel Touré University Hospital in Bamako, Mali. Observation: This was a female patient, aged 20 years old, without any particular medico-surgical ATCD or notion of taking medication, who had consulted at the end of 2021, in the Internal Medicine department of the University Hospital Gabriel Touré from Bamako for primary amenorrhea and delayed puberty. At the end of the clinical and paraclinical examination, a hypometabolism syndrome and pubertal delay were noted. On an evolutionary level, development of secondary sexual characteristics (From T1 to T3, and appearance of menstruation) after one year of treatment. Conclusion: Delayed puberty may be the consequence of hypothyroidism. Opotherapy with L Thyroxine helps restore normal pubertal development.

Mayer–Rokitansky–Küster–Hauser syndrome managed with McIndoe’s vaginoplasty: a case series and literature review

Introduction: Mayer–Rokitansky–Küster–Hauser (MRKH) syndrome is a congenital anomaly characterized by the absence of the uterus and the upper two-thirds of the vagina. It is a rare congenital anomaly with an incidence of 1 in 5000 female live births. Case series: The authors describe three cases of females presenting with primary amenorrhoea who were diagnosed with MRKH syndrome. The patients were managed with McIndoe’s vaginoplasty with neovagina creation with an amnion graft. Discussion: Management of MRKH syndrome involves vaginoplasty with neovagina creation. The approach to neovagina creation can be done surgically or non-surgically. Non-surgical creation of the vaginal cavity involves serial use of vaginal dilators, while there are several ways for surgical creation of neovagina. The modified Abbe-McIndoe procedure using amnion to create neovagina is a minimally invasive, rapid, and simple procedure with no risk of immune rejection because the amnion membrane lacks histocompatibility antigens. In addition, the graft is also readily available, storable, and inexpensive. Conclusion: Diagnosis of MRKH syndrome can be made when a young female with primary amenorrhoea and normal secondary sexual characteristics has agenesis of the uterus, and upper two-thirds of the vagina revealed on ultrasonography or magnetic resonance imaging. The patient can be offered treatment with vaginoplasty with neovagina creation.

Insulin sensitivity, body composition and bone mineral density after testosterone treatment in transgender youth with and without prior GnRH agonist therapy

Background1.8% of youth identify as transgender; a growing proportion are transgender male (female sex, male gender identity). Many receive gonadotropin releasing hormone agonist (GnRHa) therapy to suppress endogenous puberty and/or will start testosterone to induce secondary sex characteristics that align with gender identity. Objective(s)To determine the effects of 12 months of testosterone on cardiometabolic health among transgender youth, including insulin sensitivity, body composition, and bone mineral density and whether changes in outcomes differ based on prior GnRHa treatment. MethodsParticipants (n = 19, baseline age 15.0 ± 1.0 years) were examined prior to and 12 months after testosterone therapy in a longitudinal observational study. Fasted morning blood draw, a 2-hour 75-gram oral glucose tolerance test, body composition and bone mineral density (dual-energy X-ray absorptiometry) were assessed at baseline and 12 months. Insulin sensitivity was estimated by HOMA-IR and Matsuda index. Changes were compared with mixed linear regression models evaluating time (baseline, 12 months), group (GnRHa treatment yes/no), and their interaction. ResultsIn the entire cohort, fasted insulin decreased (median [25,75 %ile]: −3 [-5, 0] mIU/L, p = 0.044) and 2-hour glucose increased (mean ± standard deviation): +18.5 ± 28.9 mg/dL, p = 0.013 from baseline after 12 months of testosterone therapy. There were no significant changes in HOMA-IR (p = 0.062) or Matsuda index (p = 0.096), nor by GnRHa status. Absolute (+6.2 [4.7, 7.5] kg, p = 0.016) and percent fat-free mass increased (+7.3 [5.4, 9.1] %, p = 0.003) and percent fat mass declined (−7.4 [-9.3, 5.3]%, p = 0.005) for the entire cohort. There were time*group interactions for absolute (p = 0.0007) and percent fat-free mass (p = 0.033). There were time*group interactions for bone mineral content (p = 0.006). ConclusionsTwelve months of testosterone in transgender adolescents resulted in changes in body composition and bone mineral density, with baseline differences between the +/-GnRHa group and convergence after 12 months. There were no changes in insulin sensitivity over time or between groups.

Alterations of the hormones follitropin and lutropin in the blood of young mice dosed with lindane

BackgroundThe gonadotropins lutropin and follitropin stimulate steroid synthesis in the gonads during gonadal maturation by promoting oogenesis and spermatogenesis. Endocrine disruptors such as lindane (γHCH) can alter the reproductive stage so and could alter puberty by interfering with lutropin and follitropin signaling. γHCH was classified by World Health Organization as “moderately restricted”, and in 2009 it is production and agricultural use was prohibited, under the Stockholm Convention, however, its use as a pharmaceutical treatment against lice and scabies is still allowed (and mainly, for use by the health sector). This study aimed to examine the effects of single-dose exposure to γHCH in young mice (Mus musculus) on lutropin and follitropin concentration, and to correlate gonadal maturity status and phenotypic characteristics.MethodsYoung γHCH-treated mice (16 females and 16 males) received a single dose of 25, 10, 5, or 0 µg/ml γHCH per gram of body weight, ocular route. Once secondary sexual characteristics were observed, the gonads were dissected and examined using histological techniques and the ovarian follicles were classified as dependent and independent of gonadotropins; the testicles were classified as inactive, early and late spermatogenesis; and the blood was processed with the lutropin and follitropin ELISA kit.ResultsThe results indicate significant differences in the concentration of lutropin and follitropin between males and females (Fisher p < 0.05). The lutropin and follitropin levels showed a tendency to decrease in females, whereas, in males they tended to increase as the dose of γHCH increased. In females, it was observed that at the follitropin and lutropin decreased 42.3% and 83.7%, respectively at dose of 25 µg/ml; while in males, follitropin increased 51.9% at the dose of 25 µg/ml, and at the dose of 10 µg/ml lutropin increased by 242.5%. Contrary to that, gonadal maturity increased in females and decreased in males with increasing γHCH dose, disagreeing with the idea that gonadotropins coincide with the onset of puberty and gonadal maturity.ConclusionsThe results allow us to infer that exposure to γHCH could promote non-monotonic responses; however, this does not seem to alter puberty in the doses and conditions of the present experiment.

No immediate attentional bias towards or choice bias for male secondary sexual characteristics in Bornean orang-utans (Pongo pygmaeus).

Primate faces provide information about a range of variant and invariant traits, including some that are relevant for mate choice. For example, faces of males may convey information about their health or genetic quality through symmetry or facial masculinity. Because perceiving and processing such information may have bearing on the reproductive success of an individual, cognitive systems are expected to be sensitive to facial cues of mate quality. However, few studies have investigated this topic in non-human primate species. Orang-utans are an interesting species to test mate-relevant cognitive biases, because they are characterised by male bimaturism: some adult males are fully developed and bear conspicuous flanges on the side of their face, while other males look relatively similar to females. Here, we describe two non-invasive computerised experiments with Bornean orang-utans (Pongo pygmaeus), testing (i) immediate attention towards large flanges and symmetrical faces using a dot-probe task (N = 3 individuals; 2F) and (ii) choice bias for pictures of flanged males over unflanged males using a preference test (N = 6 individuals; 4F). In contrast with our expectations, we found no immediate attentional bias towards either large flanges or symmetrical faces. In addition, individuals did not show a choice bias for stimuli of flanged males. We did find exploratory evidence for a colour bias and energy efficiency trade-offs in the preference task. We discuss our null results and exploratory results in the context of the evolutionary history of Bornean orang-utans, and provide suggestions for a more biocentric approach to the study oforang-utan cognition.

Strategies For The Treatment Of Chronic Bronchitis In Children, Taking Into Account Sexual Development

A study was carried out to study the characteristics of sexual development and measures of secondary prophylaxis in patients with chronic bronchitis. The study included 84 children with chronic bronchitis aged 10 to 16 years. Of them, 37 (44%) are girls and 47 (56%) are boys. Moderate - severe course of the disease was revealed in 43 (51,2%) children, severe - in 41 (48,8%). It was revealed that chronic bronchitis adversely affects the sexual development of children of both sexes. The pace of development of secondary sexual characteristics occurs gradually and does not complete until the end of puberty. The comprehensive stage-by-stage secondary prophylaxis we proposed for children with chronic bronchitis disease contributed to the improvement of both the underlying disease and sexual development.

Features of the biological development of schoolchildren in the Moscow region with different levels of health

BACKGROUND: The study of the health status of children and the dynamics of the physical and biological development of schoolchildren remains an urgent scientific issue both in our country and worldwide. The lifestyles of modern children and adolescents change based on regional standards for physical and biological development. AIM: To analyze the dynamics of puberty in modern schoolchildren depending on their health status and update biological development standards of schoolchildren in the Moscow region. MATERIALS AND METHODS: In the 2020s, an observational single-center cross-sectional study of the health status and physical and biological development of schoolchildren aged 12–16 years was performed in Moscow. Physical and biological development was assessed in each age and sex group according to standard methods using certified anthropometric instruments to measure height and weight. RESULTS: The study involved 508 schoolchildren with different levels of health, who were divided into groups 1 (n=107, 21.1%), 2 (n=208, 58.7%), and 3 (n=103, 20.2%). Diseases of the eye and its adnexal apparatus (24%) were prevalent functional abnormalities, and musculoskeletal system and connective tissue diseases were found in 20% of the sample. The level of biological development in groups 1 and 2 in comparison with group 3 did not differ significantly (р 0.05); however, later development of secondary sexual characteristics tended to occur in schoolchildren with chronic diseases. Significant (р=0.038) differences in the age at menarche in girls were found depending on the level of physical development. Thus, when developing standards for biological development, data of schoolchildren in groups 1 and 2 must be considered. CONCLUSION: The results of this study clearly demonstrated the prevalence of functional abnormalities and chronic diseases of various nosologies among schoolchildren in the Moscow region. The results of the study provided an update on the standards for the biological development of schoolchildren in Moscow.

A case of Mayer-Rokitansky-Kuster- Hauser syndrome type I

Rokitansky-Kuster-Hauser (MRKH) syndrome is a rare congenital disorder characterised by absence of uterus and vagina. Ovaries and fallopian tubes are normal and functional. Its incidence is 1 in 4500-5000 female. Patients present with primary amenorrhea with normal external genitalia and development of secondary sexual characteristics, and 46XX karyotype. It is usually diagnosed by MRI, CT or transabdominal USG, with MRI being the gold standard.

Clinical features and follow-up study on 55 patients with adolescence-onset methylmalonic acidemia

Objective: To investigate the clinical features and outcomes of adolescence-onset methylmalonic acidemia (MMA) and explore preventive strategies. Methods: This was a retrospective case analysis of the phenotypes, genotypes and prognoses of adolescence-onset MMA patients. There were 55 patients diagnosed in Peking University First Hospital from January 2002 to June 2023, the data of symptoms, signs, laboratory results, gene variations, and outcomes was collected. The follow-ups were done through WeChat, telephone, or clinic visits every 3 to 6 months. Results: Among the 55 patients, 31 were males and 24 were females. The median age of onset was 12 years old (range 10-18 yearsold). They visited clinics at Tanner stages 2 to 5 with typical secondary sexual characteristics. Nine cases (16%) were trigged by infection and 5 cases (9%) were triggered by insidious exercises. The period from onset to diagnosis was between 2 months and 6 years. Forty-five cases (82%) had neuropsychiatric symptoms as the main symptoms, followed by cardiovascular symptoms in 12 cases (22%), kidney damage in 7 cases (13%), and eye disease in 12 cases (22%). Fifty-four cases (98%) had the biochemical characteristics of methylmalonic acidemia combined with homocysteinemia, and 1 case (2%) had the isolated methylmalonic acidemia. Genetic diagnosis was obtained in 54 cases, with 20 variants identified in MMACHC gene and 2 in MMUT gene. In 53 children with MMACHC gene mutation,1 case had dual gene variants of PRDX1 and MMACHC, with 105 alleles. The top 5 frequent variants in MMACHC were c.482G>A in 39 alleles (37%), c.609G>A in 17 alleles (16%), c.658_660delAAG in 11 alleles (10%), c.80A>G in 10 alleles (10%), c.567dupT and c.394C>T both are 4 alleles (4%). All patients recovered using cobalamin, L-carnitine, betaine, and symptomatic therapy, and 54 patients (98%) returned to school or work. Conclusions: Patients with adolescence-onset MMA may triggered by fatigue or infection. The diagnosis is often delayed due to non-specific symptoms. Metabolic and genetic tests are crucial for a definite diagnosis. Treatment with cobalamin, L-carnitine, and betaine can effectively reverse the prognosis of MMA in adolescence-onset patients.

Radiological study of carrying angle among male and female and its clinical importance

: The long axes of the ulna and humerus form an acute angle when the upper limb is in its anatomical position and the elbow is fully extended is known as the carrying angle. It is typically larger in females than in males Anthropologists use the carrying angle to predict a person's sex; orthopaedic surgeons use it to surgically treat a variety of elbow disorders; and total elbow prosthetics. The study aimed to assess the carrying angle in an attempt to determine its value in both sexes, proposing a simple and reliable method for measuring it.The elbow radiographs taken in the normal anteroposterior and lateral views were assessed independently and uniformly by anatomists. A total of 70 adult radiographs between the ages of 18 to 76 years, 35 males and 35 females were gathered. To measure the angle, two lines were drawn: one along the mid-axis of the upper limb of the forearm, passing between the radius and ulna through the superior radioulnar joint, and the other along the mid-axis of the lower third of the humerus, where the angle is measured.The carrying angle is statistically more significant in females than in males among 70 radiographs. The p-value was 0.000 and the mean and SD for males were 14.3±2.740, while the mean and SD for females were 21.92±3.170.The current study concludes that there is a significant gender difference between females and males; carrying angle influences secondary sexual characteristics.

Delayed diagnosis of pituitary stalk interruption syndrome in a girl with short stature, spaniomenorrhea and normal secondary sexual characteristics: a case report

Delayed diagnosis of pituitary stalk interruption syndrome in a girl with short stature, spaniomenorrhea and normal secondary sexual characteristics: a case report

Is the deficiency in secondary sex characteristics a potential manifestation of SPENCD (Spondyloenchondrodysplasia)?

Is the deficiency in secondary sex characteristics a potential manifestation of SPENCD (Spondyloenchondrodysplasia)?

Mayer–Rokitansky–Kuster–Hauser Syndrome Type II with Mullerian Duct Aplasia–Renal Dysplasia–Cervical Somite Anomalies Association: A Case Report and Review of Literature

Mayer–Rokitansky–Kuster–Hauser (MRKH) syndrome, also known as Mullerian aplasia/Mullerian agenesis, is characterized by the absence of the uterus and upper two-thirds of the vagina in women who have normal ovarian function and secondary sexual characteristics. The syndrome occurs at a rate of approximately 1 in every 4500–5000 females. Its primary clinical presentation is primary amenorrhea. MRKH syndrome varies in severity and is classified into two types: type I, which is isolated, and type II, involving abnormalities in other organ systems, particularly the kidneys and skeleton. Due to the nature of this condition, individuals with MRKH syndrome may experience significant psychological challenges, highlighting the importance of counseling. This case report describes the diagnosis of type II MRKH syndrome in a 14-year-old girl with primary amenorrhea, also associated with Mullerian duct aplasia–renal dysplasia–cervical somite anomalies.

Isolation, culture, characterization and the functional study of testicular Leydig cells from Hezuo pig.

Testosterone, an important sex hormone, regulates sexual maturation, testicular development, spermatogenesis and the maintenance of secondary sexual characteristics in males. Testicular Leydig cells are the primary source of testosterone production in the body. Hezuo pigs, native to the southern part of Gansu, China, are characterized by early sexual maturity, strong disease resistance and roughage tolerance. This study employed type IV collagenase digestion combined with cell sieve filtration to isolate and purify Leydig cells from the testicular tissue of 1-month-old Hezuo pigs. We also preliminarily investigated the functions of these cells. The results indicated that the purity of the isolated and purified Leydig cells was as high as 95%. Immunofluorescence analysis demonstrated that the isolated cells specifically expressed the 3β-hydroxysteroid dehydrogenase antibody. Enzyme-linked immunosorbent assay results showed that the testosterone secretion of the Leydig cells cultured invitro (generations 5-9) ranged between 1.29-1.67 ng/mL. Additionally, the content of the cellular autophagy signature protein microtubule-associated protein 1 light chain 3 was measured at 230-280 pg/mL. Through this study, we established an invitro system for the isolation, purification and characterization of testicular Leydig cells from 1-month-old Hezuo pigs, providing a reference for exploring the molecular mechanism behind precocious puberty in Hezuo pigs.