[Effects of ethane dimethane sulfonate on fetal Leydig cells in neonatal rats].

OBJECTIVE: To investigate the effects of ethane dimethane sulfonate (EDS) on fetal Leydig cells (FLCs) in neonatal rats. METHODS: In the study, 40 male neonatal SD rats were divided into control group and experimental groups. The rats in the experimental groups aged 3 days (PND3) were intraperitoneally injected with one single EDS (75, 100 and 125 mg/kg). The samples were collected on PND7. The body and testes was weighed, and the serum level of testosterone was detected. One testis was for histological analysis (3ß-hydroxysteroid dehydrogenase stainining), and the other was frozen in refrigerator for molecular determination (RT-PCR, Western blot). RESULTS: Compared with the control group, on day 4 after EDS treatment, significant decrease of serum testosterone was observed in the 75, 100 and 125 mg/kg experimental groups [(0.542 ± 0.117) µg/L, (0.124 ± 0.021) µg/L, (0.113 ± 0.015) µg/L, vs. (0.834 ± 0.172) µg/L, P<0.05]. Meanwhile, fetal Leydig cells in clusters disappeared with a lower expression level of Hsd3b1 and Cyp17a1 after EDS treatment in the testes of neonatal male rats in EDS (100 mg/kg)-treatment group (P<0.001). CONCLUSION: Ethane dimethane sulfonate can specifically deplete the fetal Leydig cells in testes of neonatal rats. Thus we could establish the FLCs' depletion model to know more about these testicular interstitial cells.

[Clinical characteristics of pediatric victims in the Lushan and Wenchuan earthquakes and experience of medical rescue].

OBJECTIVE: To get a more comprehensive understanding of the clinical characteristics of pediatric victims in earthquake and to summarize the experience of medical rescue. METHODS: The clinical information was collected from the pediatric victims who were admitted to West China Hospital, Sichuan University following the Lushan earthquake in 2013 and Wenchuan earthquake in 2008. The clinical data were compared between the pediatric victims in the two earthquakes. RESULTS: Thirty-four children under 14 years of age, who were injured in the Lushan earthquake, were admitted to the West China Hospital before April 30, 2013. Compared with the data in the Wenchuan earthquake, the mean age of the pediatric victims in the Lushan earthquake was significantly lower (P<0.01), and the mean time from earthquake to hospitalization was significantly shorter (P<0.01). In the Lushan earthquake, 67.6% of the injured children had variable limb fractures; traumatic brain injury was found in 29.4% of hospitalized children, versus 9.5% in the Wenchuan earthquake (P<0.05). Among the 34 children, no amputation and death occurred, and all the 13 severe cases started to recover. CONCLUSIONS: There were higher proportions of severely injured children and children with traumatic brain injury in the Lushan earthquake than in the Wenchuan earthquake. But these cases recovered well, which was possibly due to timely on-site rescue and transfer and multi-sector, multi-institution, and multidisciplinary cooperation.

[Induced differentiation of human adipose-derived stem cells into Leydig cells in vitro].

OBJECTIVE: To explore the feasibility of inducing the differentiation of human adipose tissue-derived stem cells (ADSCs) into Leydig cells in vitro. METHODS: We isolated ADSCs by digestion with Collagenase I from the subcutaneous adipose tissue, cultured them in the DMEM/F12 medium with 10% fetal bovine serum, and detected the expression of vimentin by immunohistochemistry. We exposed the ADSCs to different concentrations of human chorionic gonadotropin (HCG) for different times, determined the expression of StAR mRNA by real-time PCR, and measured the HCG-induced proliferation of ADSCs by MTT. After a week of induction by HCG and DMSO, we conducted 3beta-HSD immunohistochemistry, and detected the testosterone level in the supernatant and lysis of the cells by radioimmunoassay. RESULTS: The ADSCs grew well with a positive expression of vimentin. The expression of the StAR gene was positively correlated with the increased concentration of HCG, reaching the peak at HCG 10 U/ml in 1 week culture. The proliferation of ADSCs was significantly increased by HCG induction. A positive expression of 3beta-HSD was observed after 1 week induction with HCG 10 U/ml and DMSO 3.2 x 10(-6)mol/L. CONCLUSION: HCG enhances the expression of the StAR gene and the proliferation of ADSCs. Induced by HCG 10 U/ml and DMSO 3.2 x 10(-6) mol/L, ADSCs tend to differentiate into Leydig cells.

[In utero exposure to dichlorvos induces apoptosis of Leydig cells in rats].

OBJECTIVE: To observe the influence of the organophosphate insecticide dichlorvos on the apoptosis of Leydig cells in the male offspring of the SD rats exposed to dichlorvos, and to investigate the role of the changes of Leydig cells in genitourinary malformation. METHODS: Twenty-one pregnant SD rats were divided into a corn oil control group and 6 dichlorvos groups, the former given by gavage 1.0 ml corn oil daily, and the latter dichlorvos at the dose of 1, 4, 8, 16, 20 and 24 mg/kg daily from the 12th to 17th day of conception. After birth, 5 male neonates were randomly selected from each of the control and dichlorvos groups, and their testes were harvested to be analyzed by HE staining, immunohistochemistry with anti-caspase-3 antibodies and DAPI fluorescent staining. At 90 days after birth, another 5 of the male offspring were taken from each group and their testes were collected for the same analyses. RESULTS: Statistically significant differences were found in the number of both the caspase-3 positive and DAPI labeled Leydig cells in the testes of the rat offspring between the corn oil and the 4, 8, 16, 20 and 24 mg/kg dichlorvos groups (P < 0.05), but not between the control and the 1 mg/kg dichlorvos groups (P > 0.05). The apoptosis of Leydig cells was increased in the male offspring of the dichlorvos-exposed SD rats in a dose-dependent manner. CONCLUSION: Exposure of pregnant rats to dichlorvos can increase the apoptosis of Leydig cells in the male offspring, which, in turn, may reduce the number of Leydig cells, interfere with the testis function during the embryonic period, and damage the development of the genitourinary system.

Chordee without hypospadias: report of 79 Chinese prepubertal patients.

A series of Chinese prepubertal patients with congenital chordee without hypospadias is presented and the clinical data described. From July 1999 to September 2006, 79 boys with congenital chordee without hypospadias were treated in the Department of Pediatric Surgery, West China Hospital of Sichuan University, China. The ages ranged from 21 months to 14 years, with a mean of 76.8 months (6.4 years). The patients were categorized according to structural defect into 4 groups, with the aid of intraoperative artificial erection. Group I included those with skin tethering (28 cases, 35.4%); group II, fascial chordee (22, 27.8%); group III, corporal disproportion (10, 12.7%); and group IV, urethral tethering (19, 24.1%). Chordee-related structural defect was considered the only criterion for classification, and urethral dysgenesis influenced the choice of surgical procedure. The chordee in group I patients was corrected with penile degloving; group II, release of dense fibrous tissue in addition; group III, dorsal-midline-plication-based correction; and group IV, longitudinal-island-flap-urethroplasty-based repair. At a mean follow-up of 14.8 months (range, 2 to 63), all patients had penile straightening except 1 group III patient with residual curvature that was managed upon reoperation. Glans dehiscence occurred in 1 group II patient who underwent a tubularized incised plate urethroplasty. Urethrocutaneous fistula and urethral stricture were found in 2 group IV patients who underwent island flap urethroplasty. With the categorization based on structural defect, chordee without hypospadias may be managed well with minimized complications.

[Pathological changes in the testes of the rats with hypospadia induced by dichlorvos].

OBJECTIVE: To study the mechanism of dichlorvos leading to hypospadia of rats. METHODS: From the 12th to the 17th day of conception, 20 pregnant female rats (the experiment group) were given 10 mg/(kg x d) dichlorvos, while another 10 (the control group) administered 1.5 ml 0.9% NaCl/day. Out of 88 male newborns of the 20 experimental mother rats, 22 had hypospadia, while out of the 33 male newborns of the 10 controls, none had the problem. Five hypospadia newborns from the experiment group and another 5 normal ones from the control group were raised to sexual maturity, and then their testes were excised and embedded in paraffin, and the tissue sections were analyzed by regular HE staining and SP immunohistochemical staining with Calretinin. RESULTS: HE staining showed that the number of Leydig cells in the testis tissues of the hypospadia rats decreased significantly compared with the normal ones, but no change was observed either in the number or in the morphology of the seminiferous tubules. Moreover, the Calretinin positive Leydig cells were reduced dramatically in the testes of the hypospadia rats. CONCLUSION: Pregnant female rats, when exposed to dichlorvos, may cause reduction of testis Leydig cells in their male offsprings. Thus the probable mechanism of rat hypospadia induced by dichlorvos may lie in the decrease of the testosterone level caused by damage to Leydig cells from dichlorvos toxicity.

[Autocrine regulation for transforming growth factor-beta 1 by fibroblast in vitro].

OBJECTIVE: To explore the autocrine regulation for the transforming growth factor-beta 1(TGF-beta 1) by the fibroblasts in vitro. METHODS: Fibroblasts were cultured in vitro which isolated from the normal prepuce by circumcision. TGF-beta 1 concentration were determined with ELISA for the different passages and the different time at the same passage. RESULTS: The TGF-beta 1 concentration was different for the distinct passage of fibroblasts and achieved the peak (450 ng/L) at the sixth passage. Among the same fibroblasts, the TGF-beta 1 concentration was different in the various days and passage of the summit (680 ng/L) on the fifth day, which was 2.5 times as high as that of the first day. CONCLUSION: The fibroblasts possess the ability of autocrine of the TGF-beta 1 and have some regularity. It will provide the theory basis for the research about TGF-beta 1 regulation mechanism and the medical application about salvia miltiorrhiza.