Risk Factors for Testicular Atrophy in Children With Testicular Torsion Following Emergent Orchiopexy.

Objective: To analyze the risk factors for testicular atrophy (TA) in children with testicular torsion (TT) following emergent orchiopexy. Methods: Clinical data of patients with TT undergoing orchiopexy were retrospectively reviewed, including age at surgery, affected side, delayed surgery (12-24 h and more than 24 h), echogenicity of testicular parenchyma on ultrasonography (ETPU), testicular blood flow on Color Doppler ultrasonography (CDUS), surgical findings (intraoperative blood supply, the degree of torsion, and surgical approaches), and follow-up. The primary outcome was the rate of TA after orchiopexy. The secondary outcome was the testicular volume loss (TVL) between the affected testis and the contralateral. Results: A total of 113 patients were enrolled in this study with a median age of 11 years. The median follow-up was 21 months. Patients had a median TVL of 51.02% and 44 (38.94%) of them developed severe TA during follow-up. TA was significantly associated with age at surgery (P < 0.0001), delayed surgery (P = 0.0003), ETPU (P = 0.0001), and intraoperative blood supply (P = 0.0005). Multivariate logistic regression analysis showed that school-age children (OR = 0.069, P < 0.001) and puberty (OR = 0.177, P = 0.007) had a decreased risk of TA compared with preschool children, and that heterogeneous ETPU (OR = 14.489, P = 0.0279) and delayed surgery >24 h (OR = 3.921, P = 0.040) increased the risk of TA. Multivariate analysis demonstrated that ETPU (F = 16.349, P < 0.001) and delayed surgery (F = 6.016, P = 0.003) were independent risk factors for TVL. Conclusions: Age at surgery, delayed surgery, and ETPU may play a crucial role in predicting the TA in children with TT following emergent orchiopexy. Moreover, blood flow measured by CDUS could not predict the outcome properly.

Anatomy and management of upper moiety vascular variation in children with duplex kidney.

PURPOSE: To describe the upper pole vascular anatomy of duplex kidney and provide our experience with laparoscopic upper pole partial nephrectomy (LUPPN). METHODS: A retrospective study was performed among patients with duplex kidney who underwent computed tomography angiography at one single institution, some of whom were subsequently treated with LUPPN. According to imaging results and intraoperative findings, the arterial supply to the upper moiety of a duplex system was classified based on number and branching pattern. RESULTS: A total of 84 children were included in the study. Twenty patients (23.8%) were managed conservatively and LUPPN in lateral position was performed in the others. All laparoscopic procedures were successfully completed as planned without conversion. No major intraoperative complications occurred. Of these patients, 68 cases (73.1%) were supplied with one branch of the renal artery. The vascular anatomy of duplex kidney was classified into three patterns according to the variation of arteries. In 71 cases (76.3%), the renal artery separated into two or more arteries near the renal parenchyma, called perihilar arterial branching. Other branching patterns featured accessory renal arteries and branches of the adrenal artery. Gender was not significantly associated with the vascular number (p = 0.19) and the pattern of variation (p = 0.83). CONCLUSIONS: LUPPN is an effective technique for children with duplex kidney. The upper renal moiety is mainly supplied by one branch of the renal artery and the most common pattern is perihilar arterial branching. Determining vascular variation before surgery might be beneficial to avoid intraoperative hemorrhage and accidental vessel injury. TYPE OF STUDY: Retrospective cohort study. LEVEL OF EVIDENCE: Level IV.

Land use and land cover change in agro-pastoral ecotone in Northern China: A review.

Land use/cover change (LUCC) is the most direct manifestation of the interaction between human activities and natural ecosystems. In recent years, due to the increasing human activities, regional environment has been dramatically changed. As one of heavily influenced and fragile and vulnerable ecosystems, the agro-pastoral ecotone in Northern China is a "hot spot" for land use/cover change research. Based on a literature review of LUCC in the agro-pastoral ecotone in Northern China, we first systematically summarized the progress of the boundary definitions of the agro-pastoral ecotone, and then synthesized the current findings, methods, procedures, topics, environmental impacts and adaption of LUCC in the region. Finally, we proposed that few comprehensive, process-based ecosystem simulations and eco-environmental impact studies had been reported in the current LUCC research and called for more multi-disciplinary, multi-methods, and multi-scale researches in the future LULC research in this area.

Retrospective analysis to determine outcomes of patients with bilateral Wilms tumor undergoing nephron sparing surgery: 15-year tertiary single-institution experience.

PURPOSE: To describe our clinical experience with nephron sparing surgery (NSS) for bilateral Wilms tumor and evaluate the outcomes of patients treated at one of the largest pediatric medical centers in China. METHODS: Medical records of children with bilateral Wilms tumor undergoing NSS in the Children's Hospital of Chongqing Medical University during a 15-year period were retrospectively analyzed. Data collected were composed of age at surgery, tumor response, tumor rupture during resection, final pathologic margins, use of radiation therapy, pathology reports, renal function, and patient survival. RESULTS: A total of 18 eligible patients (10 males, 8 females) with bilateral Wilms tumor at a mean age of 2.28 ± 1.12 years were identified. The administration of preoperative chemotherapy did not result in universally successful outcomes. All children underwent successfully unilateral or bilateral NSS, of which one had positive pathologic margins and five received radiation therapy postoperatively. The rates of tumor rupture and positive lymph nodes involvement were 11.1 and 19.4%, respectively. The pathological study showed favorable histology and unfavorable histology in 32 and 4 kidneys, respectively. The 4-year event-free survival and overall survival rates were 68.18 and 85.56%. In univariable analysis, tumor histology (p = 0.0028) and disease stage (p = 0.0303) appeared significantly associated with overall survival. After a median follow-up period of 41.5 months (range 10-89), three of the surviving patients were diagnosed with hypertension and one had renal insufficiency. CONCLUSIONS: Our experience suggests that NSS has become a feasible and effective option with good oncologic outcomes. Further research, ideally in a multicenter randomized manner, is warranted to better assess the role of NSS in this challenging clinical scenario.

Diagnosis and treatment of urethral prolapse in children: 16 years' experience with 89 Chinese girls.

OBJECTIVE: To review our experience and results in the diagnosis and treatment of urethral prolapse (UP) in Chinese girls. PATIENTS AND METHODS: We conducted a retrospective chart review of 89 consecutive girls (aged <16 years) with UP and without other complications, who received treatment for UP from January 1999 to January 2015 (a study period of 16 years) at the Children's Hospital of Chongqing Medical University, China. Data analysed included: age, symptoms, clinical findings, predisposing factors, management, and outcomes. RESULTS: The presenting symptoms in the 89 girls were: mass (54 girls), bleeding (34), and dysuria/straining at micturition (one). In all, 14 patients received conservative treatment as their symptoms were mild, and 75 were successfully treated by excision of the prolapsed urethral mucosa or ligation over a Foley catheter, as their symptoms were severe and recurred too frequently to be managed conservatively. The mean postoperative length of stay for ligation was 7.76 days and for excision was 4.57 days. Ligation over a Foley catheter had a longer hospital stay. CONCLUSIONS: UP is a rare condition occurring in prepubertal girls, evidenced by a urethral mass and bleeding. Increased physician awareness and early recognition of UP avoids unnecessary examinations and patient anxiety.

A 22-year retrospective study: educational update and new referral pattern of age at orchidopexy.

OBJECTIVES: To determine the current age at orchidopexy in China and whether changing targets have altered practice, as research suggesting progressive deterioration in an undescended testis (UDT) has led to the reduction in the target age for orchidopexy to 6-12 months but it is still unknown whether changing targets have altered practice. PATIENTS AND METHODS: The demographics of orchidopexies performed in the Children's Hospital of Chongqing Medical University between 1993 and 2014 were reviewed. A survey of the general publics' awareness of UDT and survey of primary healthcare practitioners' current opinion on age at orchidopexy and referral patterns were performed. RESULTS: In all, 3784 orchidopexies were performed over 22 years. The median age at orchidopexy fell between 1993 and 2014. There was an initial drop in the median age for orchidopexy between 2000 and 2010 (36 months) compared with 1993 and 2000 (48 months) (P < 0.05); however, beyond the corresponding target age (<18 months). The age for orchidopexy between 2010 and 2014 was also beyond the corresponding target age (6-9 months). The survey of the general public showed that 0.98% had knowledge of UDT and none of them knew about the target age for orchidopexy in the survey of 5393 cases. In all, 63.46% of them were told about the UDT by healthcare practitioners at the 1-4 months postnatal baby check. Furthermore, only 2% of the healthcare practitioners knew the recommended age for orchidopexy was 6-9 months and only 14.3% of them would directly make a surgical referral to paediatric surgery specifically at this point. CONCLUSIONS: The recommended orchidopexy age is not being achieved and we recognise the national need to address this. The approach should include the right cognition of cryptorchidism among the general public and earlier primary care referral directly from the routine postnatal baby check to a specialist centre prepared to undertake surgery in this age group.

[Long-term exposure to PM2.5 from automobile exhaust results in reproductive dysfunction in male rats].

OBJECTIVE: To explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats. METHODS: Forty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 µg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 µg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression. RESULTS: The conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier. CONCLUSION: Long-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.

[Testicular teratoma in children: Analysis of 64 cases].

OBJECTIVE: To improve the diagnosis and treatment of testicular teratoma in children by analysis of clinical data. METHODS: We retrospectively analyzed the clinical data about 64 cases of testicular teratoma treated in the Children's Hospital of Chongqing Medical University from 1995 to 2014. RESULTS: Sixty-one of the cases presented painless scrotal mass with a sense of bearing down and the other 3 cases were confirmed because of empty scrotum diagnosed as cryptorchidism. The level of serum alpha fetal protein ( AFP) was obviously increased in 46 cases but normal in the other 18 preoperatively. Ultrasonography manifested abnormal inhomogeneous echo zones with calcification or necrosis. X-ray examination presented patchy or curvilinear high-density shadows in 28 cases. Forty-one of the patients underwent testis-sparing surgery (TSS) , 20 received high inguinal orchiectomy, and 3 refused surgical treatment. Pathological examination revealed 3 mature germinal layers in the 49 cases of mature teratoma and immature germinal tissue, including the original neural tube, and 11 cases of immature teratoma. The mature cases were exempted from chemotherapy, while the immature cases received the combination of cisplatin, etoposide, and bleomycin (PEB). The patients were followed up for 2 years postoperatively, which revealed no recurrence or metastasis. CONCLUSION: Most children with testicular teratoma presented painless scrotal mass with a sense of bearing down and with abnormal serum AFP in most cases. Ultrasonography and plain radiography of the scrotum contribute to the diagnosis of the tumor. TSS is the main treatment option and intraoperative frozen-section can help the surgeons decide on the surgical mode. Postoperative chemotherapy is necessitated for immature teratoma but not for mature cases.

The cyclophosphamide metabolite, acrolein, induces cytoskeletal changes and oxidative stress in Sertoli cells.

The aim of this study is to explore the mechanism by which acrolein (ACR), a metabolite of cyclophosphamide (CP), induces immature Sertoli cell cytoskeletal changes. Sertoli cells obtained from rats were cultivated and treated with 50 and 100 µM ACR. XTT assays were performed to detect cell viability. Activities of superoxide dismutase (SOD), glutathione peroxidases (GSH-Px), and catalase (CAT), as well as total anti-oxidation competence (T-AOC) were examined. Superoxide anion levels were detected by a fluorescent probe. Cell ultrastructure changes were observed by transmission fluorescent microscope. Actin filament (F-actin) distribution was detected by immunofluorescence, and ERK and p38MAPK expression were detected by western blot analysis. ACR significantly decreased the viability of Sertoli cells in a dose- and time-dependent manner. T-AOC and the antioxidant activity of SOD, CAT and GSH-Px, were decreased in ACR-treated groups compared with the control group. The levels of reactive oxygen species (ROS) in ACR-treated Sertoli cells were increased. In addition, characteristics of cell apoptosis such as mitochondrial swelling, aggregated chromatin, condensed cytoplasm, nuclei splitting, and nuclei vacuolization were observed in ACR-treated cells. Furthermore, ACR-treatment also induced microfilament aggregation, marginalization and regionalization. The expression levels of ERK and p38MAPK were also increased in ACR-treated cells in a dose- and time-dependent manner. ACR, a major CP metabolite, impairs the cytoskeleton which is likely caused by induction of the oxidative stress response through up-regulation of ERK and p38MAPK expression.

Mutation screening of BMP4 and Id2 genes in Chinese patients with congenital ureteropelvic junction obstruction.

UNLABELLED: Ureteropelvic junction obstruction (UPJO) is the most common congenital anomaly of the urinary tract. Evidence has shown that BMP4 and Id2 play crucial roles in nephrogenesis, alterations of which may cause ureteral developmental anomalies. Here, we directly sequenced the coding sequences in BMP4 and Id2 genes of 108 unrelated Chinese patients with ureteropelvic junction stenosis. One missense mutation c.485G> A (p.R162Q) in BMP4 and two synonymous mutations (c.1167T> C in BMP4 and c.108A> G in Id2) were detected in three cases. None of these variations were present in the 150 normal controls. Comparative amino acid sequence alignments of BMP4 in humans and other vertebrate orthologs show that p.R162 located to a highly conserved amino acid residue. Moreover, computational analysis predicted that R162Q probably infect the function of BMP4 protein. CONCLUSION: The mutation c.485G> A in BMP4 might be one of the causes of human UPJO. Further functional studies are required to validate the association between this variation and UPJO. Otherwise, Id2 mutations do not seem to be involved in this disease.

A rare case of simultaneous occurrence of Wilms' tumor in the left kidney and the bladder.

Wilms' tumor is the most common malignant solid tumor of the kidneys in children. Extrarenal Wilms' tumor is extremely rare. Herein, we report an 8-month-old boy with a chief complaint of frequent micturition and dysuria for 10 days. Physical examination and ultrasonography evaluation revealed simultaneous involvement of neoplasms in the left kidney and the bladder. Following excision of the masses, both were identified as Wilms' tumor by histopathology and immunohistochemistry. The two neoplasms are presumed to have developed independently because of the different pathological manifestation.

Di-(2-ethylhexyl) phthalate upregulates ATF3 expression and suppresses apoptosis in mouse genital tubercle.

OBJECTIVES: To investigate the effect of di-(2-ethylhexyl) phthalate (DEHP) on the expression of activating transcription factor 3 (ATF3) and apoptosis of fetal mouse genital tubercle (GT). METHODS: In this developmental toxicity study, pregnant C57BL/6 mice were exposed to corn oil or DEHP (100 or 500 mg/kg/day) from embryonic day 12 (ED12) to ED16. Apoptosis was characterized by Terminal transferase dUTP nick end labeling (TUNEL) assay. Using RT-PCR and western blot, the expressions of ATF3 and apoptosis-related genes (P53, Bcl-2 and Bax) were investigated. RESULTS: Apoptosis of fetal mouse GT cells notably decreased after DEHP treatment. DEHP activated ATF3 both at the mRNA and protein levels in GT. Furthermore, pro-apoptotic P53 was downregulated and the ratio of anti-apoptotic (Bcl-2)/pro-apoptotic (Bax) was not significantly changed. CONCLUSIONS: These results suggest that DEHP may induce external genital defects via a mechanism involving apoptosis, which might correlate with the regulation of ATF3 and P53 expressions.

[Hemodynamic and histological effects of unilateral testicular torsion on the contralateral testis in immature rats].

OBJECTIVE: To observe the hemodynamic and histological effects of unilateral testicular torsion on the contralateral testis in immature rats, and compare the results of different treatments. METHODS: Testicular torsion models were established in 3-week-old rats and randomized into a normal control, a testicular torsion, a reposition and an orchiectomy group. The systolic peak velocity of the right testicular artery was measured by color Doppler before and 8, 12, 24 and 72 h after the operation. Histological observations of the right testes were performed 2 h after testicular torsion, 12 h after testicular reposition and orchiectomy and when the rats were 9 weeks old. RESULTS: The blood supply of the immature right testes increased continuously after testicular torsion of the left side. Interstitial edema and ultrastructure changes were observed in the testicular torsion, reposition and orchiectomy groups. The right testis weight was significantly greater in both the testicular torsion and orchiectomy groups than in the normal control group of the 9-week-old rats (P < 0.01). No significant differences were noted in the right testicular seminiferous tubule diameter (STD) , count measure spermatogenic (CMSE) and testicular biopsy score (TBS) among the four groups (P > 0.05). CONCLUSION: Unilateral testicular torsion increases blood supply and induces histological changes in the contralateral testis in immature rats. Reposition and orchiectomy following light injury are prognostic of similar results.

[Phyllanthus urinaria extract promotes N-cadherin expression in nitrogen mustard-disrupted testicular tissues in vivo].

OBJECTIVE: To explore the protective effect of the Phyllanthus Urinaria (PU) extract on the N-cadherin expression in the testicular tissues disrupted by nitrogen mustard (HN2) in vivo. METHODS: HN2 was intraperitoneally injected into male KM mice at the dose of 5 mg/kg to make reproductive toxicity models, and at the same time PU was administered for intervention at the dose of 125 mg/kg, 250 mg/kg and 500 mg/kg. N-cadherin distribution, mRNA and protein expression in the testicular tissues were detected by immunohistochemistry, RT-PCR and Western blotting. RESULTS: N-cadherin was mainly distributed in the membrane and cytoplasm of Sertoli cells at the basement of seminiferous epithelia, Leydig cells and peritubular cells, scarcely expressed in the basement of seminiferous epithelia and peritubular cells after HN2 administration. The expressions of mRNA and proteins of N-cadherin were significantly elevated with the increased dose of PU (P < 0.01). Compared with the normal control, the distribution and expression of N-cadherin showed no significant differences in either the high-dose PU group or the HN2 with high-dose PU intervention group (P > 0.05). CONCLUSION: The PU extract can effectively promote the N-cadherin expression in the testis tissues disrupted by HN2.

[Surgical correction of buried penis in children by solution and fixation of the penile venter].

OBJECTIVE: To search for a simple and effective surgical procedure for the treatment of the buried penis in children. METHODS: We designed a new surgical procedure for the treatment of the buried penis, making a "V" incision in the venter of the radix penis, solving the superficial and deep fascia and the abnormal attachment of the dartos muscle, and fixing the left and right side of the radix penis respectively to 1 and 11 o'clock points in the fascia of the pubis nodus in the knee-chest position. A total of 38 children with the buried penis underwent the newly designed procedure and were followed up for 2-6 months. RESULTS: All the patients were discharged with desirable cosmetic appearance of the penis and completely revealed penile shafts. Follow-up showed satisfactory penile shape in 35 cases (92.11%), slightly buried penis in 2 and cicatricial constriction of the ostium praeputiale in 1. CONCLUSION: The newly designed surgical procedure of solution and fixation of the penile venter is simple and effective for the treatment of the buried penis in children and well worthy to be introduced into clinical practice.

Long-term coculture of spermatogonial stem cells on sertoli cells feeder layer in vitro.

OBJECTIVE: To construct a cell model of spermatogonial stem cells (SSCs) cocultured on Sertoli cells feeder layer in vitro, and study the proliferation characteristics of SSCs. METHODS: Sertoli cells and SSCs were separated from testes of 14-15 days and 6 days KM mice respectively by two-step enzyme digestion. SSCs were seeded on the Sertoli cells layer at 5 days in culture. The clones of SSCs on the sertoli cells layer were detected, and cast-off cells in culture medium were counted. RESULTS: SSCs began to proliferate and differentiate 24 hours after being cultured on the Sertoli cells layer, and there were a few paired (Ap) cell clones. With more time of culture, the number of Ap cell clones decreased gradually, meanwhile the number of aligned (Aal) cell clones increased, then Aal cell colonies retained stable quantity after 120 hours in culture, it could retain (51.2 +/- 5.8) days under the condition of the culture medium being changed every 4 or 5 days. CONCLUSION: SSCs can proliferate colonially on the feeder layer of Sertoli cells, and retain stable morphous and quantity. SSCs cultured on Sertoli cell feeder layer provide a cell model for studying SSCs biological behavior and interruptions of drugs or toxins on spermatogenesis in vitro. Coculture

[Effect of cyclophosphamide on spermatogonial stem cells].

OBJECTIVE: To perform a preliminary study on the dysfunction of spermatogonial stem cells (SSCs) induced by cyclophosphamide. METHODS: According to development stage of spermatogenesis in Wistar rat, 72 rats were divided into three groups, 1, 3 and 9 weeks groups, respectively. Then 24 rats per group were randomly divided into experimental and control groups, and 100 mg/kg cyclophosphamide was injected by intraperitoneal injection in experimental groups, with the same volume of normal saline in the control. Apoptosis of spermatogenic cells were detected after 24 hours by TUNEL. Then 60 one-week rats, whose germ cells were only SSCs, were randomly divided into experimental and control group. We detected c-Kit by immunohistochemistry and cell cycle by flow cytometry at 24 hours, 3 and 9 weeks. RESULTS: There was no significant difference in apoptosis of germ cells in 1 week between experimental group and control group ( P > 0.05); however, there were significant differences among other groups. The ratio of S stage and c-Kit expression of spermatogonial cells were decreased in one-week rats at 24 hours, 3 and 9 weeks in experimental group compared with the control, respectively (P < 0.01). CONCLUSION: Cyclophosphamide does not significantly induce SSCs apoptosis. It may be more important to interfere the proliferation and differentiation of SSCs.