Objective:To retrospectively evaluate the clinical effect of mitral valve repair for rheumatic mitral stenosis.Methods:We retropectively analyze the clinical datd of 50 rheumatic mitral disease patients undergoing mitral valve repair from January 2016 to March 2019, the clinical outcome was compaired with those of patients undergoing mitral valve replacement. The operation time, cardiopulmonary bypass time, blood loss, ICU time, hospital stay, and postoperative cardiac function were analyzed, and followed up for 2 years to assess mitral regurgitation, cardiac function, and complication rates.Results:The time of cardiopulmonary bypass and ascending aorta occlusion in the valve repair group were longer than those in the valve replacement group ( P<0.05), and the postoperative ventilator assistance time, ICU stay time, and hospital stay were shorter than those in the valve replacement group ( P<0.05). After 2 years of follow-up, no patients died in the two groups. The rehospitalization rate in the valve repair group was lower than that in the replacement group ( P<0.05), and there was no significant difference in the reoperation rate between the groups ( P>0.05); There was 1 case (2%) of moderate mitral valve regurgitation in the mitral valve repair group, no moderate or severe mitral valve stenosis, no paravalvular leakage in the mitral valve replacement group, and no significant difference between the two groups ( P>0.05). The left ventricular end-diastolic diameter and left ventricular ejection fraction in the mitral valve repair group were significantly better than those in the mitral valve replacement group ( P<0.05). Conclusion:Mitral valve repair is effective in treating rheumatic mitral stenosis. It is beneficial to protect heart function, reduce postoperative anticoagulation complications, and does not increase the rate of reoperation. It is a safe, effective and feasible treatment.

Objective To investigate the proper procedure for repairing different urethrocutaneous fistulas after primary urethroplasty for hypospadias.Method There were 101 cases,whose age ranged from 27 months to 171 months (mean 61 months),underwent urethrocutaneous fistula repairing secondary to the primary hypospadiasis urethroplasty from January 2010 to December 2015.The methods of the repairing were chosen mainly on the site and the size of the fistula,which included three types.The coronal fistula with a thin band of tissue stretching between the glans wings was classified as type Ⅰ (n =24).For the rest of the small fistulas at penis coronal ditch and penis body,the small fistula (diameter ＜ 3 mm) was classified as type Ⅱ (n =57) and the large fistula (diameter ≥3 mm) was classified as type Ⅲ(n =30).The urethroplasty was performed in the type Ⅰ cases.The ligation and transfixion was performed in type lⅡ cases.And the tension free repairing with continuous suture was performed in type Ⅲ cases.De-epithelization dartos fascia flap or tunica vaginalis flap covering was performed in all cases.After removing the catheter,all cases were followed-up at least 1 year.The successive operation was termed as no complication,such as urethral stricture,urethral diverticulum and urethrocutaneous fistula.Result Totally 111 fistulas were repaired by the methods described above.The mean followed-up duration was 32 months (ranging 12-48months).Total recurrence of fistula was 11.7％ (13/111) in different type fistulas,including 16.7％(4/24) in type Ⅰ,3.5％ (2/57) in type Ⅱ and 23.3％ (7/30) in type 1Ⅲ.Conclusions Different classification and treatment of urethrocutaneous fistula after urethroplasty by the site and size of fistula can improve the outcome of the operation.It is necessary to cover the tissue tightly to a new urethra during the operation.

BACKGROUNDTo clone DNA sequence of human telomerase reverse transcriptase (hTERT) promoter, and study its transcriptional activities in various human lung cancer cells and normal lung cell.

METHODShTERT promoter of 1.1 kb was amplified with polymerase chain reaction (PCR) method, utilizing DNA of 293 cell as template. After DNA sequencing with correct result, the hTERT promoter was inserted into luci-ferase reporter vectors (pGL3-Basic) to reconstruct a recombinant plasmid named pGL3-hTERTp. Then the reconstructed plasmid was transiently transfected into lung cancer cell lines A549, SPC-A-1, LTEPa-2, NCI-H446, YTMLC, GLC-82, A2 and human embryonic pulmonary fibroblast cell line MRC-5. The transcriptional activities of hTERT promoter in various cells were determined by measuring the luciferase activities after 48 hours of transfection.

RESULTSElectrophoresis demonstrated that cloned hTERT promoter was about 1.1 kb, and DNA sequencing showed a same sequence as registered in GenBank. The cloned hTERT promoter was located between 1 126 bp and 43 bp in upstream of the transcription start site ATG and the length was 1 086 bp. The recombinant plasmid pGL3-hTERTp was confirmed by double digestion and PCR method with correct results. Luciferase assay showed there were different transcriptional activities of hTERTp in various lung cancer cell lines, but not in the MRC-5 cell line.

CONCLUSIONSThe hTERT promoter cloned in this study has transcriptional activities in various lung cancer cell lines but not in normal cell. It may act as control element in tumor-targeting gene therapy with hTERT.

Objective To investigate the effect of replacing a smaller-size catheter after tubularized incised plate (TIP) urethroplasty on postoperative urethral complications.Methods The data of 116 hypospadias patients underwent TIP urethroplasty performed by the same urologist in our hospital from January 2014 to December 2017 were retrospectively analyzed.The age of patients ranged from 0.5-15.4 years,with the median age of 1.5 years.Meatal location was distal in 47 (39.5％),midshaft in 62 (52.1％) and proximal in 10(8.4％) patients.Unhealthy urethral plate occurred in 49 (42.2％) patients.F8 catheter was used for urethroplasty in 92 (79.3 ％) patients,F10 in 17 (14.7％),and F12 in 7 (6.0％).According to the pattern of urinary diversion,patients were divided into two groups.Conventional catheter group in 56 patients (group A):the catheter was chosen in as large size as possible for urethroplasty without tension.Replace a smaller-size catheter group in 60 patients (group B):replace a smaller-size catheter after urethroplasty on the basis of group A.There were no statistically significant differences in age,meatal location and catheter size between the two groups (P ＞ 0.05).Results The mean follow up was 23 months (range 6-66 months).There were 73(62.9％) cases of orifice overflow during indwelling catheterization.There were 5 (4.3％) patients needed further reoperation result from postoperative urethral complications,including 4 cases of fistula and 1 case of meatal stenosis.The process of replacing a smaller-size catheter after urethroplasty in group B was simple and without adverse reactions.There were 15 (26.8％) cases of orifice overflow in group A and 58 (96.7％) in group B.There were statistically significant differences in orifice overflow between the two groups (P ＜ 0.05).There were 5 (8.9％) cases of postoperative urethral complications in group A and 0 case in group B.There were statistically significant differences in postoperative urethral complications (P ＜ 0.05).Conclusions The process of replacing a smaller-size catheter after TIP urethroplasty in was simple and subsequently contributed to less postoperative urethral complications.The results of replacing a smaller-size catheter after TIP urethroplasty were superior to that of no replacing conventionally.

Objective To explore the change of urine proteins from nephrostomy tube and renal function in patients with very severe hydronephrosis after nephrostomy in order to determine the treatment strategy of either renal preservation or resection.Methods Retrospective analysis of clinical data from March 2015 to March 2017 of 28 cases with congenital unilateral severe hydronephrosis in children's hospital affiliated to Zhejiang University,including 19 boys and 9 girls,with 20 cases on the left and 8 on the right,an average age of 26.6 (0.3 to 122.0) months at the first consultation and 2 cases of urinary biochemical abnormalities.Front to rear diameter for puncture were between 5.7 and 7.6 cm,with an average of 6.7 cm,and the differential renal function (DRF) for the affected kidneys were 6.98 ％-9.89 ％ (average 8.65 ％) before puncture.The children underwent percutaneous renal perforation fistula.According to the recovery of renal function after perforation fistula,those who underwent nephrectomy were recruited as a nephrectomy group and the children whose kidney were preserved as kidney preservation group.Comparison 24 hours before and 3 months after puncture was made beteween groups regarding the proportion of urine,pH of the renal pelvis urine,oαl microglobulin (α1-MG),albumin (Alb),β2 microglobulin (β2-MG),immunoglobulin G (IgG),transferrin (TRF),and the recovery of the damaged renal function (direnal function DRF).Results Twenty-eight cases of percutaneous nephrolithotomy were successful without puncture complications,with 10 cases in the nephrectomy group and 18 cases in the renal preservation group.The difference of renal pelvis urine from the affected kidney of the nephrectomy group showed no statistical significance 24 hours before and 3 months after the procedure when comparing the following parameters,including urinary specific gravity (1.006 ± 0.007 vs.1.009 ± 0.005),pH value (7.74 ± 0.41 vs.7.70 ± 0.32),urine creatinine value [(462.20 ± 158.01) μmol/L and (449.20 ± 143.41) μmol/L],αl microglobulin [(14.03 ± 4.360) mg/L vs.(13.84 ± 2.55) mg/L],β2 microglobulin [(0.76 ± 0.28) mg/L vs.(0.65 ±0.14) mg/L],immunoglobulin G [(1 292.20 ± 303.50) mg/L vs.(1 175.33 ± 376.67) mg/L],and kidney function (DRF) [(8.86 ±0.67) ％ vs.(9.90 ± 1.26)％].While microalbumin [(8 644.40 ± 829.54) mg/L vs.(3 145.10 ± 1 445.02) mg/L] and transferrin [(445.70 ± 46.71) mg/L vs.(214.30 ± 40.13) mg/L,P ＜ 0.05] were significantly increased 3 months after than 24 h before the procedure in nephrostomy group.There were no significant differences of the urine specific gravity 24h before and 3 months after the procedure in the kidney preserved group (1.003 ± 0.003 vs.1.005 ± 0.003,P ＞ 0.05).The creatinine value of 3 months after the procedure was significantly increased than that 24 hours within the procedure [(654.50 ± 154.52tμmol/L) vs.(423.94 ± 172.74) μmol/L],and the urine pH (7.28 ± 0.32 vs.7.91 ± 0.56),α1 microglobulin [(5.85 ± 0.38) mg/L vs.(12.58 ± 3.40) mg/L],microalbumin [(571.50 ± 167.14) mg/L vs.(2 343.28 ± 576.22) mg/L],β2 microglobulin [(0.14 ±0.05)mg/L vs.(0.53 ±0.13) mg/L],immunoglobulin G [(247.38 ±75.29) mg/L vs.(1 026.44 ±245.42)mg/L],and transferrin [(67.64 ± 16.34) mg/L vs.(249.17 ±78.62) mg/L] were significantly decreased 3 months after than 24 hours before the procedure.The renal function was higher 3 months after than 24 hours within the procedure [(8.53 ± 0.80) ％ vs.(20.50 ± 7.87) ％,P ＜ 0.05].Conclusions The increase of creatinine value and the decrease of urine pH,α1 microglobulin,microalbumin,β 2 microglobulin,immunoglobulin G and transferrin in renal pelvis urine suggested that renal function recovered after renal puncture,which is important for next-step plan of treatment strategy.

Objective To investigate the effect of a modified preputial flap urethroplasty in twostage treatment of severe hypospadias.Methods The clinical characteristics of the severe hypospadias patients (41 cases) who underwent the staged urethroplasty by using the procedure of preset urethral plate with preputial flap from January 2015 to December 2016 were analyzed retrospectively.We used a modified method (modified group,23 cases):Form the distal urethra with the transected distal urethral plate by using tubularize incised plate (TIP) procedure during the first stage operation,after completely straightened the penis,urethral plate was preseted with transverse preputial flap at the penis shaft.While in the traditional group (18 cases),urethral plate was preseted with preputial flap by using Bracka procedure after transecting urethral plate.The corresponding missing part of urethra underwent urethroplasty at the second stage operation six months later.The patients in the modified group were 9 to 18 months old,and the median age was 13 months;meatus were located at the penis shaft in 10 cases,scrotum in 12 cases,perineum in 1 case.The patients in the traditional group were 9 to 18 months old,and the median age was 13 months;meatus were located at the penis shaft in 6 cases,scrotum in 10 cases,and perineum in 2 cases.There was no statistic difference in age and meatus position between the two groups.Results During the first stage operation,distal urethra was repaired by 13-19 mm,with an average of (14.5 ± 1.3) mm,and the proximal urethra was repaired by 0-6 mm,with an average of (3.1 ± 2.4) mm at the modified group.While at the traditional group,the length of proximal urethra was repaired of 0-9 mm,with an average of (5.6:±:2.9) mm.The urethral length required for reconstruction was measured during the second stage operation,with an average of (26.3:t:4.4) mm in the modified group and (40.5 ± 3.3) mm in the traditional group (P ＜ 0.05).There were 3 case of urethral stricture after removed the catheter,with 2 cases in the modified group and 1 case in the traditional group.Postoperative follow-up was 2 to 3 years,with an average of 2.4 years.There were 3 cases (3/23,13.0％) of urethral fistula in the modified group and 3 cases (3/18,16.7％) in the traditional group.Ascended testis occured in 2 patients in the traditional group after operation.No case of urethrostenosis,diverticulum,chordee or concealed pennis was recorded.There was no significant difference in postoperative complications between the two groups (P ＞ 0.05).Conclusions The modified staged preputial flap method shorten the new forming urethra by making full use of its own materials at the second stage operation,which was helpful to reduce complications.

Objective To review the clinical characteristics of urethral prolapse in female children and summarize our experience of treatment.Methods A retrospective analysis of the clinical characteristics of 102 patients with urethral prolapse from January 2007 to December 2017 was conducted at The Children's Hospital of Zhejiang University School of Medicine.The age of the patients ranged from 8-156 months with an median of 80 months.The presenting symptoms in the 102 girls were:bleeding in 57 patients (55.9％),mass in 31 patients (30.4％),and dysuria/urinary frequency,urgent and pain in 14 patients (13.4％).In all,58 patients were managed conservatively with Sitz baths as their masses were small,39 underwent prolapse reduction under topical anesthesia and Sitz baths because their mass were large,and 5 patients were treated by excision of the prolapsed urethral mucosa with four-quadrant excisional technique because thrombosed urethral prolapse at first visit.Results A total of 89 patients were cured after conservative treatment (87.3％),8 patients were converted to surgical treatment because frequent recurrence with conservative treatment.No urethral stricture,active hemorrhage and recurrent were found in 13 patients after operation.Conclusions The most common clinical manifestations of urethral prolapse are urethral mass and bleeding.Most patients can be cured by conservative treatment.The patients whose symptoms were severe or suffered from frequent recurrence of urethral prolapse should be managed with surgical excision.

Objective To investigate the application of pedicled tunica vaginalis flap cover new urethra in redo and second-stage hypospadias repair.Methods Retrospective analysis was performed in 45 redo hypospadias repair cases,aged from 1 year 3 months to 9 years 4 months (median 5 years 3month).All patients were divided into urethral dehiscence after hypospadias repair (group A) and the second stage surgery of two-stage hypospadias repair (group B) based on medical history.According to the site of dehiscence,the group A were further divided into midshaft urethral dehiscence (A1) and glans and distal urethral dehiscence with urethral opening located on midshaft (A2).After new urethra was completed,the appropriate size of the pedicledtunica vaginalis flap was transferred to cover the new urethra.Results There was no scrotal hematoma occurred in all 45 cases,and somecomplications occurred,including scrotal swelling in 1 case,high-riding testicle in 2 cases,penile clockwise torsion in 1 case,urethrocutaneous fistula in 6 cases.All the 45 cases were followed up for 1 to 4 years,with mean of 1.8 years.One more urethrocutaneous fistula occurred in group A2 and 1 more urethrocutaneous fistula with urethral stricture occurred in group B.There was no penile curvature and urethral diverticulum occurred in all cases.Conclusions Pedicled tunica vaginalis flap could be used as waterproofing layer to cover new urethra in redo hypospadias repair and presented a low incidence rate of postoperative complications.

Disorders of sex development (DSD) refer to a group of diseases characterized by abnormal congenital development of chromosomes, gonad or genitals with different pathophysiological changes and clinical manifestations. DSD is more common in neonates and adolescents, and neonates often show genital abnormalities while adolescents show abnormal sexual development during puberty. It is the international consensus that the scope of DSD should include basic clinical evaluation (internal and external genitalia and endocrine hormones), diagnostic confirmation (chromosome, genetic diagnosis), psychological assessment for children and family, treatment (sex assignment, hormone replacement and surgical intervention), potential fertility protection and long-term follow-up, which require the expertise of pediatric endocrinology, pediatric urology, clinical psychology, genetic disciplines, medical images and other related disciplines; that is, individualized management of children with DSD requires an experienced multidisciplinary team (MDT). This article reviews the recent progress on the evaluation, diagnosis and management of disorders of sex development.
Consensus ; Disorders of Sex Development ; diagnosis ; therapy ; Humans ; Medicine ; trends ; Sexual Development

Objective:To investigate the effect of microgravity-mediated Notch1 signaling on macrophage polarization on bone homeostasis.Methods:The animal model was constructed by tail-limb suspension(HLS)to simulate the microgravity environment.The animals were grouped into Control group,HLS group,HLS+NC group,HLS+si group,HLS+rhNF-κB group.ELISA was used to detect the content of TNF-α and IL-1β in serum of rats.TUNEL staining was used to detect the apoptosis of bone tissue.Immunofluo-rescence was used to detect the polarization of macrophages in bone tissue.The rat osteoblast CP-R091 microgravity model was con-structed by simulating the microgravity environment with a rotating wall bioreactor.The cell experiments were divided into Control group,HLS group,HLS+NC group,HLS+si group,HLS+rhNF-κB group.CCK-8 test was used to detect the proliferation activity of cells in each group,and AO test was used to test the apoptosis rate of cells in each group.PCR was used to detect the expression of os-teogenesis-related genes in bone tissues and cells.Western blot was used to detect the expression of Notch1,hair division-related en-hancer-1(HES-1),and Notch pathway ligand 1(Jagged1)in bone tissues and cells of each group.Results:Compared with control group,the contents of TNF-α and IL-1β in the serum of the rats in the HLS group,the apoptosis rate,and the proportion of M1 macro-phages were significantly increased.Compared with HLS group,the HLS+si group could obviously partially reverse the change trend of the above parameters,while HLS+rhNF-κB group significantly changed the above parameter values.Compared with control group,the proliferation activity of the cells in the HLS group was significantly reduced,and the apoptosis rate was significantly increased.Com-pared with HLS group,the HLS+si group could obviously partially reverse the change trend of the above parameters,while the HLS+rhNF-κB group made the above parameter values worse.The expressions of the osteogenesis-related genes collagen type Ⅰ(COL1),osteocalcin(OCN)and Runt-related transcription factor 2(RUNX2)in bone tissues and cells in the microgravity environment were significantly decreased,while the expressions of Notch-1,Hes-1 and Jagged1 were significantly increased,and the differences were statistically significant(all P<0.05).Conclusion:Microgravity-mediated Notch1 signaling regulates M1/M2 polarization of macro-phages,participates in cell proliferation and apoptosis in bone tissue,and affects the progress of bone homeostasis.