Poor outcome in congenital mesoblastic nephroma with TPM3::NTRK1 fusion: a case report from multi-disciplinary treatment to molecular tumor board.

Background: Congenital mesoblastic nephroma (CMN) is a rare renal tumor with good prognosis in children; however, cellular CMN is a special subtype with poor prognosis. The ETV6 fusion gene has been found in some cellular CMNs, whereas CMNs with TPM3::NTRK1 fusion gene have not been reported. This study aims to share the progression and treatment of a case of CMNs with TPM3::NTRK1 fusion gene, in order to provide experience for the diagnosis and treatment of such specific diseases. Case Description: We report a case of CMN with TPM3::NTRK1 fusion gene and a 3-year course of disease that originated during the fetal period. The child experienced rapid tumor progression 22 months after birth, followed by tumor recurrence 3 months after complete resection of CMN. Although traditional chemotherapy could not prevent the tumor progression. The tropomyosin receptor kinase (TRK) inhibitor larotrectinib resulted in significant inhibitory effects on metastatic lesions in the lungs, liver, and peritoneum. However, the patient ultimately died as the tumor became resistant to larotrectinib. Conclusions: CMN, is a rare pediatric renal tumor that warrant prompt surgical management. A watchful waiting approach may allow for aggressive growth of metastatic disease, as seen in this case of cellular CMN with TPM3::NTRK1 fusion gene, TRK inhibitors can play significant roles in the treatment of CMN with TPM3::NTRK1 fusion gene, but we still need to pay attention to the phenomenon of drug resistance to larotrectinib caused by site mutations of TRKA.

Comparison of proximal and distal laparoscopic ureteroureterostomy for complete duplex kidneys in children.

OBJECTIVE: To compare efficacy of proximal and distal laparoscopic ureteroureterostomy (UU) for complete duplex kidneys in children. METHODS: Patients who underwent laparoscopic UU for complete duplex kidneys between December 2016 and July 2022 were reviewed retrospectively. 71 patients who had normal lower pole moiety without vesicoureteral reflux (VUR) were recruited. All of them underwent ultrasound, voiding cystourethrography (VCUG), renal scintigraphy, and magnetic resonance urography preoperatively. Proximal laparoscopic UU was performed in 35 patients and distal laparoscopic UU in 36 patients. Double J stents were placed in normal lower pole moieties. Clinical data, including general information, diagnosis, surgical management, imaging characteristics, clinical symptoms and postoperative complications (classified according to the modified Clavien-Dindo classification), and length of stay were recorded. Measurement date comparisons between groups were performed by t test, counting date were analyzed by chi-square test. RESULTS: The study consisted of 71 patients (56 females and 15 males) with complete duplex kidneys (41 in left kidney and 30 in right kidney). The patients' mean age was 34 m (range 3-161 m) and follow-up ranged from 25 to 81 m. No significant difference was found in age and follow-up time between the two groups. Laparoscopic UU was performed in all patients successfully. The operation time of the two groups was 108.42 ± 26.95 min for distal UU vs 121.46 ± 35.15 min for proximal UU(p = 0.14). No significant difference in postoperative complications was seen between the two groups (22.2% vs 31.4%, p = 0.345). However, in terms of the grading of postoperative complications, the proximal UU group had a higher grade (3 of them had a grade of IV) and more serious complications. CONCLUSIONS: There was no significant difference in the overall incidence of complications between distal and proximal UU. Compared with proximal laparoscopic UU, distal laparoscopic UU is easier to perform with less injury to the peripheral tissues. Postoperative complications of proximal UU are more serious and more difficult to manage. We recommend complete duplex kidney ureteral reconstruction with distal UU.

Clinical features and treatment options for pediatric adrenal incidentalomas: a retrospective single center study.

BACKGROUND: The aim of this study was to investigate the clinical features and treatment options for pediatric adrenal incidentalomas(AIs) to guide the diagnosis and treatment of these tumors. METHODS: The clinical data of AI patients admitted to our hospital between December 2016 and December 2022 were collected and retrospectively analyzed. All patients were divided into neonatal and nonneonatal groups according to their age at the time of the initial consultation. RESULTS: In the neonatal group, 13 patients were observed and followed up, and the masses completely disappeared in 8 patients and were significantly reduced in size in 5 patients compared with the previous findings. Four patients ultimately underwent surgery, and the postoperative pathological diagnosis was neuroblastoma in three patients and teratoma in one patient. In the nonneonatal group, there were 18 cases of benign tumors, including 9 cases of ganglioneuroma, 2 cases of adrenocortical adenoma, 2 cases of adrenal cyst, 2 cases of teratoma, 1 case of pheochromocytoma, 1 case of nerve sheath tumor, and 1 case of adrenal hemorrhage; and 20 cases of malignant tumors, including 10 cases of neuroblastoma, 9 cases of ganglioneuroblastoma, and 1 case of adrenocortical carcinoma. CONCLUSIONS: Neuroblastoma is the most common type of nonneonatal AI, and detailed laboratory investigations and imaging studies are recommended for aggressive evaluation and treatment in this population. The rate of spontaneous regression of AI is high in neonates, and close observation is feasible if the tumor is small, confined to the adrenal gland and has no distant metastasis.

pH-responsive, self-sculptured Mg/PLGA composite microfibers for accelerated revascularization and soft tissue regeneration.

Biodegradable Mg/polymer composite fibers offer a promising therapeutic option for tissue injury because of bioactive Mg2+ and biomimetic microstructure. However, current studies are limited to the contribution of Mg2+ and the single microstructure. In this study, we designed Mg/poly (lactic-co-glycolic acid) (Mg/PLGA) composite microfibers that significantly enhanced angiogenesis and tissue regeneration synergistically by Mg2+ and self-sculptured microstructure, due to spontaneous in situ microphase separation in response to the weakly alkaline microenvironment. Our composite microfiber patch exhibited superior performance in the adhesion, spreading, and angiogenesis functions of human umbilical vein endothelial cells (HUVECs) due to the joint contribution of the hierarchically porous microstructure and Mg2+. Genomics and proteomics analyses revealed that the Mg/PLGA composite microfibers activated the cell focal adhesion and angiogenesis-related signaling pathways. Furthermore, the repair of typical soft tissue defects, including refractory urethral wounds and easily healed skin wounds, validated that our Mg/PLGA composite microfiber patch could provide favorable surface topography and ions microenvironment for tissue infiltration and accelerated revascularization. It could cause rapid urethral tissue regeneration and recovery of rabbit urethral function within 6 weeks and accelerate rat skin wound closure within 16 days. This work provides new insight into soft tissue regeneration through the bioactive alkaline substance/block copolymer composites interactions.

Modified onlay island flap versus tubularized incised plate urethroplasty for hypospadias reoperation.

This study aimed to assess the outcomes of modified onlay island flap (MOIF) urethroplasty for hypospadias reoperation. A retrospective analysis was conducted using clinical data on children reoperated with MOIF or tubularized incised plate (TIP) urethroplasty for hypospadias in the Children's Hospital of Nanjing Medical University (Nanjing, China) from December 2016 to September 2020. A total of 90 children with hypospadias who failed primary repair and were reoperated on with MOIF (MOIF group, n = 46) or TIP urethroplasty (TIP group, n = 44) were recruited. All children were postoperatively followed up for at least 6 months. Six patients with postoperative urinary fistula, 2 patients with glans dehiscence, and 1 patient with urethral diverticulum were reported in the MOIF group, while 7 patients with postoperative urinary fistula, 2 patients with urethral stricture, and 1 patient with glans dehiscence were reported in the TIP group. Postoperative infection, flap necrosis and other complications were not recorded. A total of 37 (80.4%) patients in the MOIF group and 34 (77.3%) patients in the TIP group were successfully reoperated. Although the success rates of MOIF and TIP urethroplasty are comparable, pediatricians should master different types of repair techniques for individualized hypospadias reoperation. MOIF urethroplasty can be recommended after failure of the primary repair.

A novel onlay urethroplasty for hypospadias with mild chordee after degloving: modified for complete removal of scar tissue underlying the urethral plate and for long-term outcomes.

Urethral plate (UP)-preserving urethroplasty is simple and has few complications, but it may affect the development of penis in the long term and lead to recurrent chordee. In this study, we used obliquely cut UP to repair hypospadias with mild chordee after degloving (15°-30°) and compared the results with onlay urethroplasty to explore its rationality and feasibility. Between April 2018 and October 2020, 108 hypospadias patients underwent onlay urethroplasty or modified onlay urethroplasty. Clinical data were prospectively collected, and medium-term outcomes were assessed at follow-up. The complications were compared between the two groups. Forty-four patients underwent the modified onlay procedure (Group I), with follow-up time (mean ± standard deviation [s.d.]) of 23.2 ± 4.5 (range: 17-31) months. Sixty-four patients underwent a standard onlay procedure (Group II), with follow-up time (mean ± s.d.) of 39.7 ± 3.9 (range: 32-46) months. There was no difference in age at surgery. The urethral defect length and operative time were longer in Group I. Six cases of fistula and one case each of stricture and diverticulum were reported in Group I. In Group II, 11 cases of fistula and one case each of stricture and diverticulum were reported. The complication rates were 18.2% and 20.3% in Group I and Group II, respectively (P > 0.05). These medium-term follow-up results demonstrate that the modified onlay procedure (oblique cut UP urethroplasty) is a safe and feasible technique for hypospadias with mild chordee after degloving. Compared with standard onlay urethroplasty, this modified procedure is conducive to the complete removal of scar tissue underlying the UP without increasing the risk of surgical complications.

Comparative study of one-stage and the novel two-stage urethroplasty using the transected urethral plate and transverse preputial island flap (TPIF) for severe hypospadias: a single-center experience.

BACKGROUND: To evaluate a novel two-stage method of urethroplasty using a transected urethral plate and transverse preputial island flap (TPIF) for proximal hypospadias with severe chordee. METHODS: We retrospectively analyzed 91 cases of proximal hypospadias, who had undergone either one-stage or two-stage urethroplasty. Comparisons between the two groups were made based on age, glans diameter, the length of the urethral defect, and the rate of complications. RESULTS: There was no significant difference in age at surgery, glans diameter, and the length of urethral defect between the two groups. In the one-stage group, 18 cases of urinary fistula and one case each of glans dehiscence, urethral diverticulum, and urethral diverticulum with concurrent glans dehiscence, were reported. All patients were treated using urinary fistula repair or urethroplasty. In the novel two-stage group, no cases of urinary fistula were reported after the secondary urethroplasty. However, 4 cases showed dehiscence at the glans, 2 were treated through reoperation but the 2 other 2 patients reoperation. The success rate was 62.50% and 88.57% in the one-stage and two-stage groups, respectively (P<0.05). CONCLUSIONS: Two-stage urethroplasty using a tubularized urethral plate and TPIF can significantly reduce the incidence of urethral fistula of the proximal hypospadias with severe chordee. However, it may increase the number of operations that need to be performed on children who do not need staged surgery. Although we could not prove it through this study, we believe that the necessity of two-stage urethroplasty should be determined based on the development of the glans, the severity of penile curvature, and the length of the urethra defect.

Patient transfer influences the prognosis of pediatric patients operated for testicular torsion: a single-center experience.

BACKGROUND: Testicular torsion (TT) remains an emerging issue for pediatric patients. This study's objective was to determine risk factors of TT outcomes in the Chinese pediatric cohort from a single center. METHODS: We performed a retrospective cohort study of patients who met the inclusion criteria in our center from 2013 to 2018. Clinical demographics, and TT-related data, including laterality, torsion degree, torsion direction, position to the cavity of tunica vaginalis, enorchia, and outcomes, were reviewed and extracted from medical charts and databases. Statistical analysis was conducted to compare and figure out the risk factors of TT outcomes. RESULTS: A total of 84 pediatric patients were enrolled in this study. Among these variables, age, duration of symptoms, and the patient transfer were significantly associated with the outcomes of TT (P<0.005). Patients transferred from other medical facilities underwent more orchiectomy than those without transfer (univariate analysis, P<0.0001; multivariate analysis, P=0.001). No difference was found in other variables between patients with or without transfer (P>0.05). Moreover, transferred patients tended to show worse testicular function. CONCLUSIONS: We have reported that age, duration of symptoms, and patient transfer were three significant risk factors in this cohort. Patients with transfer tended to suffer from more orchiectomy and worse prognosis of TT.

[Application of the lateral preputial fascial island flap in hypospadias reoperation].

Objective: To evaluate the application and effect of the lateral preputial fascial island flap (LPFIF) in hypospadias reoperation. METHODS: We retrospectively analyzed the clinical data on the children patients undergoing hypospadias reoperation with LPFIF in our Department of Urology from December 2016 to June 2019. RESULTS: A total of 85 cases were included in this study, including 18 cases of LPFIF, 19 cases of Duplay technique, 25 cases of tubularized incised plate urethroplasty (TIP) and 23 cases of Mathieu urethroplasty. The patients were aged from 18 months to 12 years and 8 months, averaging 4.6 years. Postoperatively, all the children were followed up for ≥ 6 months, which found satisfactory appearance of the penis body and glans, the urethral orifice in the normal position of the glans, and the external orifice of the urethra fissured. Urethral fistula occurred in 2 cases and glans dehiscence in 1 after LPFIF; 5 of the patients presented urethral fistula after Duplay; 3 developed urethral fistula and 1 urethral stricture after TIP; 6 showed urethral fistula and 2 glans dehiscence after Mathieu urethroplasty. No postoperative urethral stricture, urethral diverticulum or flap necrosis occurred in any of the cases. Hypospadias reoperation succeeded in 83.3% (15/18) of the cases after LPFIF. The urine flow curve of the LPFIF cases was bell-shaped or high flat-shaped, with a maximum urinary flow rate of 8.56 ± 3.99 ml/s and an average urinary flow rate of 5.23 ± 2.32 ml/s, not significantly different from those of the TIP and Duplay cases. CONCLUSIONS: There is no statistically significant difference in the success rate of hypospadias reoperation between TIP and LPFIF. Urethroplasty with LPFIF can be used as one of the surgical options for hypospadias reoperation.

Use of double-J ureteric stents post-laparoscopic pyeloplasty to treat ureteropelvic junction obstruction in hydronephrosis for pediatric patients: a single-center experience.

OBJECTIVES: We aimed to investigate the safety concerns associated with placing double-J ureteric stents post-laparoscopic pyeloplasty surgery for congenital ureteropelvic junction obstruction (UPJO) and hydronephrosis. METHODS: A total of 1349 patients with postoperative double-J stent placement at our center were included. Clinical variables for enrolled patients were collected by two independent authors. We compared clinical variables and the efficacy of stenting post-laparoscopic pyeloplasty. RESULTS: The mean age of the patients was 4.23 ± 2.39 years. A total of 58.49% of patients were diagnosed with left UPJO with hydronephrosis and 33.95% were diagnosed with right UPJO. Furthermore, 7.56% of patients had bilateral UPJO. In all cases, 96.96% of indwelling double-J stents were successfully removed 4 weeks post-surgery. A total of 3.04% of the patients still required further management, including stent migration to the renal pelvis (0.37%), stent migration to the bladder (0.30%), prolapse of the stent through the ureter (0.15%), blockage of stents (1.85%), and fouling of stents (0.37%). CONCLUSIONS: Double-J ureteric stents used after laparoscopic pyeloplasty for treating UPJO in hydronephrosis for pediatric patients is a safe, feasible, and beneficial method, which can be recommended for routine procedures. However, caution should be practiced for follow-up and removal using this method.

Risk factors affecting the prognosis of urinary tract infection in Chinese pediatric patients: Single-center experience.

BACKGROUND: Urinary tract infection (UTI) is common in children. The purpose of this retrospective study was to determine the various risk factors that usually affect the prognosis of UTI in children diagnosed with the disease. METHODS: In the present retrospective study, we enrolled all pediatric patients diagnosed with UTI and hospitalized between 1 January 2013 and 31 July 2016 at Nanjing Children's Hospital. We then collected all the relevant patient clinical demographics and characteristics. RESULTS: The study involved 2,092 pediatric patients diagnosed with UTI. On logistic regression analysis, factors that could affect the prognosis of pediatric UTI were complications, hospitalization, intensive care unit (ICU) admission, type of UTI, urine culture results, blood lymphocytes, urine nitrites (NIT) and antibiotics (unstandardized coefficients, 0.06, <0.001, -0.28, 0.32, <0.001, 0.01, -0.11, 0.01, respectively, all P < 0.001). CONCLUSION: Complications, hospitalization, type of UTI, urine culture results, blood lymphocytes, and antibiotics had a significant, positive association with UTI prognosis. Meanwhile, ICU admission and urine NIT had a negative association with prognosis.

Comparative study of urethroplasties to reduce urethral strictures in patients with severe hypospadias.

OBJECTIVE: Transverse island pedicle flap (TIPF) plus transected urethral plate-preserving urethroplasty is increasingly used for treatment of severe hypospadias. We aimed to reduce the occurrence of urethral strictures in patients undergoing such procedures. METHODS: Sixty-five patients with severe hypospadias were enrolled. Thirty-two patients underwent onlay-tube-onlay urethroplasty (Group A), and 33 patients underwent modified Duplay urethroplasty (Group B). Postoperative complications were recorded, including fistulas, urethral strictures, and diverticula. RESULTS: Three patients (9.4%) in Group A and 10 patients (30.3%) in group B had urethrocutaneous fistulas. Three patients (9.4%) in Group A and 0 patients (0%) in Group B had urethral strictures. No patient in the two groups had symptoms of diverticulum or penile chordee. The results of uroflowmetry were better in Group B than Group A, when comparing uroflow patterns. CONCLUSIONS: TIPF plus transected urethral plate-preserving urethroplasty can lower the occurrence of stricture, which is a challenging complication. The occurrence of stricture was lower in patients who underwent modified Duplay urethroplasty, and neourethral function and quality were better in these patient. Thus, this modified procedure can be used for treatment of severe hypospadias.

A new modification of the Duckett technique for one-stage repairing urethral plate transected hypospadias: Another option for severe hypospadias?

OBJECTIVES: The study aimed to evaluate the new modification of the Duckett technique in decreasing the incidence of urethral strictures for urethral plate transected hypospadias and also explored its clinical application. METHODS: Thirty-three patients (aged 7 months to 12 years, mean age 2.8 years) who underwent repair of primary hypospadias using the new one-stage urethroplasty were enrolled. Clinical data, including cosmetic and its complications, and uroflowmetry data were documented. Uroflowmetry data of 19 patients who underwent Duckett urethroplasty were used as a comparison. RESULTS: The length of the urethral defect ranged from 2.5 to 5.0 cm. The postoperative follow-up was 14-30 months. Ten patients (30.3%) had fistulas; no patients had strictures or diverticula. All ten fistulas were small (<0.5 cm) and repaired with fistula repairing operation. The appearance of the penis remained satisfactory, and the meatus was located in the normal anatomic position. Among 17 patients who underwent uroflowmetry, all patients were bell-shaped or platforms, and Q max was 7.37 ± 2.45 ml/s. Compared with 14 of 19 patients who underwent Duckett urethroplasty, the urethral function achieved with new one-stage urethroplasty was significantly better (p < 0.05). CONCLUSIONS: The incidence of strictures was dramatically lowered in patients with proximal hypospadias. Small fistulas are common complications and can be repaired easily. Based on the uroflow pattern results, the quality of neourethra and function of it were better than Duckett urethroplasty. These preliminary results suggested that the modified procedure seems to be reliable and can be a suitable option for proximal hypospadias.

Unilateral congenital giant megaureter with renal dysplasia compressing contralateral ureter and causing bilateral hydronephrosis: a case report and literature review.

BACKGROUND: Congenital giant megaureter (CGM) is uncommon in the pediatric population. The major clinical presentations are marked protruberances and abdominal cysts. CASE PRESENTATION: We reported a case of CGM with almost the whole left ureter dilation accompanied with a 1 cm stricture at the entrance of the bladder and renal dysplasia, immediately compressing the contralateral ureter and causing bilateral hydronephrosis for the first time. At one-stage of the operation, a left nephrostomy with a right ureterolysis were performed, and a poor left kidney function was found. Then, the left kidney and ureter were cut off by nephroureterectomy at the second-stage. Eventually, the follow-up showed that the patient recovered well by abdominal ultrasound. CONCLUSION: Based on the findings of these reported literatures, CGM is rare. The physical and imaging examinations are essential for the diagnosis of CGM, and the appropriate treatment methods should be performed based on patients' specific condition.

Retrospective analysis of individual risk factors for urethrocutaneous fistula after onlay hypospadias repair in pediatric patients.

BACKGROUND: To retrospectively identify the individual risk factors for the urethrocutaneous fistula (UCF) in pediatric patients after hypospadias repair (HR) with onlay island flap urethroplasty. METHODS: A total of 167 patients who underwent primary HR at Nanjing Medical University Affiliated Children Hospital from January 2009 to December 2012 were enrolled. Clinical data including the patient' age at HR, hypospadias type and urethral defect length were documented. RESULTS: Among 167 patients, 12.6% patients (n = 21) developed UCF after HR. Postoperative UCF occurred in 3.9% (3/76) cases at age of 0-2 years, 14.3% (9/63) at 2-4 years, 20.0% (2/10) at 4-6 years and 38.9% (7/18) at 6-12 years. The incidences of UCF were 12.0% (3/25), 11.4% (5/132) and 30.0% (3/10) for distal, middle and proximal types of hypospadias. As to the urethral defect length, the incidences of UCF were 8.2% (5/61) in patients with a length of ≤ 2 cm, 12.8% (9/70) in 2-3 cm, 22.6% (7/31) in 3-4 cm and 0% (0/5) in above 4 cm. Older age at HR was significantly associated with the high incidence of UCF formation (P = 0.004), while the hypospadias type and urethral defect length did not affect it (P = 0.264 and P = 0.312, respectively). CONCLUSIONS: The patient' age at HR was a risk factor for the UCF formation after HR, and treatment of HR within two years old might be with the least incidence of UCF.

[Comparisons of efficacy and safety of tolterodine and oxybutynin in children with idiopathic overactive bladder].

OBJECTIVE: To compare the efficacy and safety of tolterodine and oxybutynin in the treatment of idiopathic overactive bladder in children. METHODS: A total of 204 children with idiopathic overactive bladder were randomly divided into three groups (n=68 each): placebo, tolterodine-treated and oxybutynin-treated. The efficacy and safety were evaluated two weeks after treatment. RESULTS: The effective rate was 25% in the placebo group, 89% in the tolterodine-treated group, and 92% in the oxybutynin-treated group. The effective rate in the two treatment groups was significantly higher than that in the placebo group (P<0.05). There was a similar efficacy between the two treatment groups. The incidence of adverse events in the tolterodine-treated group (28%) was significantly lower than that in the oxybutnin-treated group (57%) (P<0.05). CONCLUSIONS: Tolterodine has a similar efficacy to oxybutynin in the treatment of idiopathic overactive bladder in children, with better safety in pharmacotherapy.

[Effects of Di (2-ethylhexyl) phthalate on the testis and testicular gubernaculum of fetal KM mice].

OBJECTIVE: To explore the effects of Di (2-ethylhexyl) phthalate (DEHP) on the testis and testicular gubernaculum of fetal KM mice in vivo and to investigate the mechanism of DEHP-induced cryptorchidism. METHODS: Thirty healthy pregnant KM mice were randomly and equally divided into a blank control group, a corn oil control group and a DEHP group. The pregnant mice in the latter group were exposed to DEHP by gavage at the dose of 500 mg/kg body weight per day from gestation day 12 (GD12) through gestation day 19 (GD19). The effects of DEHP were observed on the number of fetuses per pregnancy, the ratio of male to female pups, the weight of the testis, the morphology and location of the testis and gubernaculum, the relative testis-bladder neck distance (TBD) and cranial suspensory ligament (CSL) residual. The expressions of the androgen receptor (AR), estrogen receptor (ER) and actin and proliferating cell nuclear antigen (PCNA) in the gubernaculum were detected by immunohistochemistry. RESULTS: DEHP reduced the testis weight and TBD, induced different degrees of testis maldescent, but produced no obvious effect on the body weight, the number of fetuses per pregnancy, the sex ratio and the testis gubernacular morphology. Under the light microscope, hypotrophy was seen in all the testis seminiferous tubules, spermatogenic cells and Sertoli cells, marked Leydig cell hyperplasia was noted, and the positive expression of AR in the gubernaculum was decreased in the DEHP group (P < 0.01). CONCLUSION: DEHP could cause dysfunction of the testis gubernaculum via its anti-androgen effect, induce cryptorchidism, and cause dysplasia and dysfunction of Sertoli cells, Leydig cells and spermatogenic cells in fetal mice.

[Di(2-ethylhexyl) phthalate affects the testes and leydig cells of neonatal KM mice].

OBJECTIVE: To explore the effects of di(2-ethylhexyl)phthalate (DEHP) on neonatal mice's testes and Leydig cells in vivo. METHODS: Pregnant mice were exposed to DEHP at the dose of 100 mg/kg, 200 mg/kg or 500 mg/kg (body weight) per day by gavage from gestation day 12 (GD 12) through postnatal day 3 (PND 3), respectively. The testis and body weights, testicular histopathology and the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) of the neonatal mice were investigated. RESULTS: The body and testis weights of the male mice's offspring were significantly reduced following DEHP exposure. Leydig cell morphology was affected significantly by DEHP as compared with the controls. Leydig cells obviously increased in the neonatal mice's testes on PND 15 and PND 30 when exposed to DEHP (500 mg/[kg x d]). Activities and positive area of the steroidogenic enzymes 3beta-HSD immunoexpression decreased markedly when exposed to DEHP (100 mg/[kg x d] or 200 mg/[kg x d]). Image analysis showed a decrease in the activities of 3beta-HSD in the animals exposed to DEHP (500 mg/[kg x d]), but an increase in the positive area of 3beta-HSD immunoexpression as compared with the control animals on PND 15 (P < 0.01). CONCLUSION: DEHP affects the Leydig cell morphology, the activity of 3beta-HSD, the testis and body weights and the testicular histopathology of neonatal mice, and it may function as an antiandrogenic agent.

[Isolation, cultivation, identification and functional study of fetal mice Leydig cells in vitro].

OBJECTIVE: To explore the methods of isolation, cultivation, purification, identification of the fetal mice testis Leydig cell and to observe its biological characteristics in vitro. METHODS: Leydig cells were isolated by 0.03% collagenase (type I) from fetal mice testis and cultured in DMEM/F12 medium. The identity and purity of Leydig cell were assessed by 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD). Cell viability was measured by trypan blue. Testosterone level in the medium of cultured Leydig cells was measured in various culture phases and cell density by radioimmunoassay. RESULTS: The purity of Leydig cell was (45.10 +/- 1.66)% before culture, and (81.17 +/- 2. 32)% 72 h after culture. The level of testosterone secreted by Leydig cells could be detected in the medium and its level was associated with the density and time of cultured Leydig cells. The secretion capacity of testosterone by single Leydig cell decreased gradually during the culturing period. CONCLUSION: The fetal Leydig cells isolated from fetal mice testis have high purity. It can be cultured and kept the secretion ability of testosterone for a few days in vitro. This system can provide a valuable model for further study on the cellular function of the Leydig cells of fetal mice.

[Histopathological changes of the cryptorchid testis and epididymis of mice exposed to DEHP].

OBJECTIVE: To investigate the effects of DEHP on the incidence of cryptorchidism and the histopathological changes of the testis and epididymis in mice. METHODS: Forty KM mice were divided into 5 groups randomly: normal control group (n=8), corn oil group (n=8), DES group (n=8), low DEHP group (n=9), and high DEHP group (n=7). DEHP[100 or 500 mg/(kg x d)], DES[100 microg/(kg x d)] and corn oil were administrated respectively to the pregnant mice from gestation day 12 (GD 12) until postnatal day 3 (PND3). The incidence of cryptorchidism with testicular morphologic and histological changes in the male pups was observed. RESULTS: Significantly increased incidence of cryptorchidism was seen in the male pups with increasing exposure to DEHP. Cryptorchid testicular volumes and weights significantly decreased compared with the control. Abnormal pathological and ultrastructural alternations of seminiferous epithelium and Leydig cells were also observed. Meanwhile less or no sperm was found in the lumen of the epididymis. CONCLUSION: During the critical period of male sexual differentiation, exposure to DEHP may lead to a significantly high incidence of cryptorchidism, testicular damage and fertility impairment in the male offspring. The effects of DEHP are dosage-dependent.