Objective To summarize the phenotypic features of an unrecognized leukoencephalopathy in infants sharing same clinical features,and to better understand the disease and provide new evidence for identification of new leukoencephalopathy. Methods Clinical and follow-up data of 13 patients with unrecognized infantile leukoen-cephalopathy were collected from Peking University First Hospital from January, 2006 to December, 2014. Results (1) There were 7 male and 6 female. The average age of onset was 11 months (4-25 months). Thirty-eight percent (5/13 cases) of patients had incentives before the onset;all of the cases had acute onset and rapid motor function regression. Fifteen percent (2/13 cases) of the patients suffered from seizures in the course of the disease. Patients′condition became stable,and cognition and motor function improved gradually 1 month after onset. No patient died till the last follow-up. (2) Imaging features:magnetic resonance imaging (MRI) of the patients was characterized by im-plicating deep white matter,presenting T1 hypointense,T2 and fluid attenuated inversion recovery ( FLAIR) hyperin-tense in the periventricular area. All of MRI showed massive and symmetric lesions with heterogeneous signal and cystic degeneration. DWI showed patch or massive hyperintense in some of the lesions. The follow-up MRI showed the original lesions decreased in 88% ( 8/9 cases ) of patients, and white matters atrophied in 55% ( 5/9 cases ) of patients;the cystic degeneration still existed and even expanded;DWI showed regional linear or spot hyperintense in 88% (8/9 cases) of patients,which was smaller than before,and distributed around the original lesions. Conclusions The patients with leukoencephalopathy caused by unknown pathogenic gene were much likely to be mitochondrial leukoencephalopathy. This study provided evidence for further exploration of new pathogenic genes causing leu-koence-phalopathy.

ObjectiveTo investigate the effects of diet-induced obesity on the developmental process of testes in pubertal Sprague Dawley (SD) rats and explore the possible reversibility. MethodsSixty one-month-old male SD rats were randomly divided into a control group ( n =10) and a model group ( n =50 ), which were fed on a normal diet and a high-fat diet, respectively. After 8 weeks, all the rats in the control group and 10 rats randomly picked out from the model group were killed. The serum testosterone and estradiol levels were measured by enzymelinked inununosorbent assay. Their left testes sections were stained by HE method, and the histology was observed under optical microscope and the spermatogenic activity was evaluated by Johnsen scoring system. The remaining 40 rats in the model group were further randomly divided into 3 subgroups: continued high-fat diet subgroup ( n =13), resume normal diet subgroup (n =13), and weight-loss subgroup (n =14). The continued high-fat diet subgroup was fed by high-fat diet, while the other two subgroups were fed by normal diet. Rats in weight-loss group took normal diet with running 20 min/d. After 6 weeks, the same parameters were assessed using the same methods. ResultsAfter 8 weeks, compared with the control group, the testosterone level of the model group significantly decreased (P =0.024) and the estradiol level significantly increased ( P =0. 017). The result of HE staining showed that the spermatogenic cell layers decreased, with part of seminiferous tubule experiencing atrophy.The number of Leydig cell also decreased and lipo vacuole was seen in the interstitial tissue of testis. The Johnsen score of the model group was significantly lower than that of the control group (P =0.000). The testosterone level was significantly lower in the continued high-fat diet subgroup than that in resume normal diet subgroup ( P =0.001 ) and weight-loss subgroup ( P =0.000), and was significantly lower in resume normal diet subgroup than that in weight-loss subgroup ( P =0.001 ). The estradiol level was significantly higher in continued high-fat diet subgroup than that in resume normal diet subgroup ( P =0.001 ) and weight-loss group ( P =0.000 ), and was significantly higher in resume normal diet subgroup than that in weight-loss group ( P =0.001 ). HE staining showed that, pathological changes aggravating and worsening compared with the control group, the model group had significantly decreased seminiferous tubule cell layers, with some seminiferous tubules experiencing atrophy.The Johnsen score was significantly higher in weight-loss subgroup than that in the other two subgroups ( P =0.000and 0.001, respectively). The Johnsen score was negatively correlated with body weight ( r =- 0.962, P =0.000), and positively correhted with the serum testosterone level ( r =0.916, P =0.000 ). Conclusions High-fat diet can induce pubertal obesity in male SD rats, which is featured by testicular hypoplasia, decreased spermatogenesis, and endocrine dysfunction. Physical exercise may improve the conditions. The degree of obesity may be negatively correlated with the spermatogenic function.

ObjectiveTo evaluate the effect of inlay buccal mucosa graft with tubularized incised urethral plates (TIP) urethroplasty for hypospadias repair. MethodsFrom January 2005 to December 2010,a total of 343 cases of hypospadias underwent a buccal mucosa graft with TIP urethroplasty.The patients aged from 6 months to 61 years,mean 7 years.One hundred and forty-seven were primary surgery cases,and 196 cases had a history of failed surgery.In primary cases,124 were distal type and 23 were proximal type.There were 137 distal type cases and 59 proximal types in failed case group.A buccal mucosa graft was harvested from lower lip,fixed into the incised urethral plates in primary cases.In re-do cases,open urethral and remove scar tissue were necessary. ResultsThe width of the buccal mucosa ranged 0.5 - 2.5 cm,length ranged 1.0 - 8.5 cm.All the patients were followed up for 6 - 36 months,average 16 months.In primary group,fistula occurred in 14 cases (9.5％ ),and urethral stricture in 2 cases (1.4％),in which 1 proximal case occurred both with urethral stricture and fistula; the overall success rate was 89.8％.In re-do group,fistula occurred in 32 cases ( 16.3％ ),and stricture in 13 cases (6.6％),in which 1 distal and 5 proximal cases occurred with both urethral stricture and fistula; the overall success rate was 80.1％. ConclusionsInlay buccal mucosal graft with TIP technique is an effective instrumentality for hypospadias repair because of its high successful rate,good cosmetic effect and without affecting on oral appearance and function in donor site.

Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.

Objective To evaluate if strategic urethrostomy could reduce complications in complicated hypospadias repair.Methods From January 2016 to August 2018,165 patients of complicated hypospadias were reviewed according to inclusion criteria.They were divided into three groups of one-stage repair (group A,n =86),two-stage repair using Bracka procedure (group B,n =49) and strategic urethrostomy (group C,n =30).The median age was 26 months in group A,24 months in group B and 28 months in group C.The median length of urethral defect was 3.0 (2.0-10.0) cm,4.0 (2.5-10.0) cm and 3.8(2.5-11.0) cm in the different three groups,respectively.No difference showed in age or length of urethral defect among 3 groups.Three groups were compared with rates of urethral fistula,urethral stricture and urethral diverticulum.Results After average of 20.0 months follow-up,the urethral stricture incidence of group C [3.3％ (1/30)] was significantly lower than that of group A [22.0％ (19/86),P =0.023] and group B[24.5％ (12/49),P =0.032].The complications were found in 7 patients with urethral fistula and 11 patients with urethral diverticulum in group A,4 patients with urethral fistula and 3 patients with urethral diverticulum in group B,none patient with urethral fistula nor urethral diverticulum in group C,respectively.The incidence of urethral fistula was 8.1％ (7/86),8.2％ (4/49) and 0 in the three groups,and the rate of urethral diverticulum was 12.8％ (11/86),6.1％ (3/49) and 0,respectively.None difference was shown neither in the incidence of urethral fistula nor urethral diverticulum among the three groups(P ＞ 0.05).Conclusions Strategic urethrostomy is a novel and effective method for complicated hypospadias repair.Application of strategic urethrostomy can significantly decrease urethral stricture and improve success rate in complicated hypospadias repair.

A reliable UPLC-MS method was developed for the simultaneous determination of 4 chemicals (Sudan Ⅰ, tetrabromobisphenol A, tris ( 1, 3-dichloroisopropyl ) phosphate and tris ( chlorisopropyl ) Phosphate) in children products. The samples were ultrasonic extracted with acetonitrile, and then the four chemicals were separated on a C18 column in 3 min. Results showed that the limit of quantification of the method was between 5 and 500μg/kg. The calibration curves were linear within 2-3 orders of magnitude with typical correlation coefficient above 0 . 9995 . The recoveries ranged from 83 . 7% to 97 . 8% with three addition levels. The sensitivity, recovery and selectivity of the method could fully meet the requirements of practical work.

AIM:We investigated how AT 1-R stimulated by mechanical stresses induces cardiac fibrosis .METHODS:We produced in vivo cardiac pressure overload model in angiotensinogen knockout ( ATG-/-) mice and in vitro mechanically-stretched cell model in cultured neonatal cardiac cells of ATG-/-mice both lack the participation of Ang II .RESULTS: Pressure overload for 4 weeks in ATG-/-mice induced myocardial hypertrophy accompanied by the significant interstitial fibrosis , however , the TGF-β, a key regulatory factor of fibrosis, was not significantly increased in these ATG-/-mice.Meanwhile, the inhibitor for AT1-R significantly inhibited mechani-cal stress-induced cardiac fibrosis in these ATG-/-models whereas inhibition of TGF-βdid not.CONCLUSION:The results showed that mechanical stress-induced fibrotic responses through AT 1-R required the phosphorylation of Smad 2 but not the involvement of TGF-β.

Objective: To reveal the clinical and genetic features of neonatal/infantile epileptic disorders caused by KCNQ2 mutations and to provide a clue for the treatment and prognosis evaluation. Methods: Twenty-two patients were collected in the Department of Pediatrics, Peking University First Hospital from April 2007 to July 2016.The phenotype-genotype analysis was conducted of the neonatal/infantile epileptic patients in whom a KCNQ2 mutation was identified by the targeted next generation sequencing. Results: Twenty-two de novo KCNQ2 missense mutations from 22 patients with neonatal/infantile epileptic disorders were found.These patients had an onset of epilepsy in early infancy (median age: 2 days). The seizure type of the first onset was mainly focal seizure.Atypical absence epilepsy, a novel phenotype of KCNQ2 mutation-induced epilepsies was found.The mortality of these patients was high, as 5 patients of the 22 patients died in the follow-up period, 4 of which might result from sudden unexpected death in epilepsy.In the 22 patients, 8 patients with anti-epileptic monotherapy became seizure-free.Of the 8 patients with a monotherapy, 3 patients were treated with valproic acid and no clinical onset was observed. Conclusions This study expands the phenotype of KCNQ2-related epileptic disorders.These patients have high mortality.Valproate acid is the potentially effective monotherapy for these patients.

Objective:To evaluate the safety and effectiveness of the model repair strategy for preputial defect in hypospadias surgery.Methods:From February 2017 to December 2022, 59 children in our hospital with an average age of (6.9±3.2) years were retrospectively analyzed. All of them were children with multiple failed hypospadias operations. According to the condition of penile scrotum skin, different methods of penile skin reconstruction were selected. Among them, 20 patients underwent penile skin flap reconstruction with simple foreskin and multiple small incision reduction, 22 patients with penile and scrotal transposition underwent penile lateral scrotum flap to complete ventral penis coverage, and 12 patients underwent middle scrotum flap with pedicled flap to cover ventral urethra. The penis was covered by inferior epigastric artery perforator flap in 5 patients.Results:The wound healed completely one week after the operation and no obvious scar formation was observed. The ventral flap of the penis was covered by the lateral scrotal flap of the penis, and no flap necrosis and wound infection were observed. The ventral urethra was covered with pedicled skin flap in the middle scrotal suture. The flap survived without scrotal hematoma. The inferior epigastric artery perforator flap was used to cover the patients with penile defect, all the flaps survived, and no complications such as wound infection and abdominal hernia occurred. No urethral fistula or urethral diverticulum was reported in all the patients. Urethral stricture occurred in 1 case of scrotal suture pedicled flap group and 1 case of lateral penis scrotal flap group, which was cured after expansion. One case with small incision and one case with lateral scrotal skin were cured after secondary repair by Mathieu method.Conclusions:For children with a history of multiple hypospadias operations, coverage of ventral skin defects of the penis is essential to reduce complications and obtain good appearance. This study summarized four effective strategies for covering penile defect from simple to complex. The small incision was used to reduce the expansion of all flaps. The lateral penile scrotal flap is more suitable for patients with penile scrotal transposition. The pedicled flap of scrotal suture and inferior superficial artery perforator flap are suitable for the repair of larger defect area.

Objective:To investigate the clinical efficacy of labial mucosa graft coupled preputial island flap urethroplasty for recurrent penile curvature after hypospadias repair.Methods:The data of 35 patients with recurrent penile curvature after hypospadias repair who underwent labial mucosa graft coupled preputial island flap urethroplasty in four hospitals from January 2021 to March 2023 were retrospectively reviewed. The median age of the patients was 15(13, 18)years old, five of them had penile dehiscence, and four had urethrocutaneous fistula. All patients received labial mucosa graft coupled preputial island flap urethroplasty, the penile flexion was confirmed by artificial erection test, and the urethra is obliquely cut at the most obvious curvature of the penis. By removing the ventral fibrotic scar tissue, the length of the urethral defect was measured. The urethral plate was reconstructed with the same length of labial mucosa graft and an adjacent preputial island flap was transplanted to cover the reconstructed urethra plate to form a new urethral lumen. The inner urethral stent tube was extracted three days after the operation, and the outer urethral stent tube was removed three weeks after the surgery. Diagnostic urethral exploration was performed 5 weeks post-operation, uroflowmetry examination was performed seven weeks after the procedure, and the perioperative and postoperative follow-up data were analyzed.Results:All patients underwent a successful surgery, with the average duration of the procedure being (114.7±20.1)minutes. After mean follow-up of (15.5±5.9)months, no penile curvature occurred, and 32 patients were successfully repaired. Uroflowmetry examination was achieved in 21 patients at the seventh week after surgery showed their mean maximum flow rate was (18.4±3.7)ml/s. Three cases (8.6%, 3/35) developed urethrocutaneous fistula, who were repaired successfully 6 months after surgery.Conclusions:Utilizing the combination of labial mucosa grafts and preputial island flaps for urethroplasty has been proven to be a reliable and safe option for individuals with recurrent penile curvature following hypospadias repair, with a high success rate and few complications.