Objective To investigate the treatment options of clomiphene(CC) resistant polycystic ovary syndrome (PCOS) related infertility. Methods Figty nine PCOS patients with CC resistant anovulation were accepted and treated by the following three protcols :(1)follicle stimulating hormone (FSH) group, 49 cycles; (2)FSH +pulsatile gonadotropin releasing hormone(GnRH) group, 13 cycles; (3)conventional in vitro fertilization (IVF)_embryo transfer(ET) group, 19 cycles. Suppressive treatment on serum luteinising hormone (LH)and testosterone (T) levels was given in the first and second groups in advance. Serum estradiol level; pregnancy rate; miscarriage rate, ovarian hyperstimulation syndrome (OHSS) and multiple pregnancy rate were compared among the three groups. Results The pregnancy rate in FSH group, FSH+GnRH group and IVF ET group werte 37%,54% and 22% respectively. The highest rate and multiple pregnancy rate was found in IVF ET group. Conclusion The rationale treatment options for CC resistant PCOS related infertility was the addition pre suppressive treatment, low dose FSH stimulated regimen subsequent pulsatile GnRH infusion. IVF were only accepted after failure with gonadotropin therapy.

Objective To study the clinical features of complicated upper urinary tract infection in children,aiming to provide a reference for clinical diagnosis and treatment of the disease.Methods The clinical data of 68 cases with complicated upper urinary tract infection hospitalized at the Department of Urinary Surgery NO.1,Children's Hospital of Shenzhen between January 2013 and August 2015 were retrospectively analyzed.Results A total of 68 cases,in which 44 were male,24 were female,and repeated infections were found in 15 cases.Fever was the most common clinical manifestation(48 cases,70.59％),while frequent urination and odynuria were rare(9 cases,13.24％).A total of 57 strains had been cultured form the submitted specimens which were obtained from the 68 cases,including 41 strains of gram-negative bacteria,15 strains of gram-positive bacteria and 1 strain of fungus.Two different strains were cultured in 3 children.Fourteen strains of Escherichia coli had 11 extended spectrum beta lactamases (ESBLs)-positive strains.In the 11 strains of klebsiella pneumoniae,8 trains were ESBLs-positive.The drug resistant rates of gram-negative bacteria to Ampicillin and Cefuroxime sodium were both more than 90％.The sensitive rates to Piperacillin/Tazobactam was more than 90％.Thirty-one cases of complicated upper urinary tract infection were cured by administering Piperacillin/Tazobactam,while 15 cases were cured by changing Cefuroxime sodium to Piperacillin/Tazobactam according to the drug sensitivity results.Conclusions Clinical manifestations of complicated upper urinary tract infection are untypical,and fever is the most common symptom.Repeated infection is common.The gram-negative bacteria is the dominant pathogen causing the complicated upper urinary tract infection.ESBLs-positive bacteria accounts for high proportion.The drug resistance rate to penicillin and the first,second generation of the cephalosporin is high.The drug sensitive rate of piperacillin tazobactam is high,with good prognosis.

[Objective] This study compared outcomes of in vitro maturation (IVM) and in vitro fertilization (IVF) intracytoplasmic sperm injection (ICSI) cycles after IVM of immature germinal vesicle (GV) oocytes.[Methods] ICSI was performed on metaphase II (MII) oocytes retrieved in 163 IVF-ICSI cycles (group I;n ＝ 987) or matured from GV stage oocytes in IVF-ICSI ( group II;n ＝ 132) and 37 IVM cycles ( group III;n ＝ 235).Fertilization and cleavage rates and embryo quality were compared among the three groups.[Results] The fertilization rate,cleavage rate and top quality embryos rate were higher in group I than group II and group III (84.9%,98.1%,and 61.6%;72.0%,90.5% and 22.1%l;75.3%,94.4%,and 25.1%,respectively).Blastomere numbers and morphology scores were highest in group I (P ＜ 0.05),but no significant differences existed between group II and group III.[Conclusion] The morphology of embryos developed from in vivo MII oocytes was superior to those from in vitro matured MII oocytes.No significant difference was observed in embryo morphology from immature GV oocytes in IVF and IVM cycles.

AIM: To explore the activation effect of calcium ionophore A23187 on unfertilized human mature oocytes after conventional in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHODS: Thirty-seven unfertilized mature oocytes from IVF and 41 after ICSI were included in our experiment. They were incubated in 5 ?mol/L calcium ionophore A23187 for 5 minutes. Second polar body extrusion and pronuclear formation were recorded 12-16 hours later. The activated oocytes were cultured for another 2 days in vitro. RESULTS: Activation rate of unfertilized oocytes from conventional IVF and ICSI were 64.9%(24/37)and 73.2%(30/41), respectively. Among 41 unfertilized oocytes after ICSI treated with A23187, 30 were activated and 24 had 2 polar body (PB) and 2 pronuclear (PN). But for the unfertilized oocytes from conventional IVF only 20% activated oocytes had 2 PN and 2 PB. The percentage difference of oocytes containing 2 PB and 2 PN between the two groups was significant ( P

AIM: To investigate the optimal materials and culture system of human primordial germ cells (PGCs) in order to establish human embryonic germ (EG) cell lines. METHODS: Human embryos of different gestational age were collected to isolate human PGCs. The isolated human PGCs were cultured in different medium and on different feeder layers, then their growth, proliferation and differentiation in different culture systems were observed. RESTILTS: The formation rate of primary colonies was higher when human PGCs were obtained with enzyme-mechanical method from 8-and 9-weeks gestational age human embryos than that from 7-weeks. Human PGCs grew better and maintained undifferentiating when mouse embryonic fibroblast or STO cells served as feeder layers and in conditional medium with hLIF, hbFGF, hSCF. CONCLUSION: 8-and 9-week gestational age human embryo are optimal material for isolating human PGCs. Enzyme-mechanical method is simple and available to isolate human PGCs. Feeder layer and growth factors are necessary for human PGCs culture in vitro.

Objective Try to determine the relationship between blastocyst quality,the clonal growth of inner cell mass(ICM)and the establishment of human embryonic cell line. Methods Coculture D 3 discarded embryos with mouse embryonic fibroblast cells(MEFs).Then remove trophoectoderm by immunosurgery after getting different quality blastocysts.Culture ICM and passage these cells on MEFs. Results Human embryonic stem cells derived from good quality blastocysts could be passaged further than that from poor quality blastocysts,and ICMs growing fast could be passaged more quickly and efficiently.Conclusion The establishment of human embryonic stem cells is closely related with blastocyst quality and the original growth of ICM.

Objective To investigate influence of chromosomal translocations on early embryo development and to evaluate the efficacy and feasibility of preimplantation genetic diagnosis (PGD)techniques through clinical analysis on PGD cycles. Methods Embryo development, efficacy of PGD and clinical outcome of 100 cycles were studied retrospectively, including 23 cycles with Robertsonian translocations, 19 cycles with reciprocal translocations, and 58 cycles for α-Thalassaemia. Results Among 354 embryos biopsied by PGD for translocations, 321 (90. 7% ) presented fluorescence in situ hybridization (FISH) results. The rate of normal/balanced embryos in the Robertsonian translocation was 38. 3% (64/167),which was significantly higher than 20. 8% (32/154) in the reciprocal translocation group. Amplification was achieved in 443 blastomeres from 537 embryos in Thalassaemia group, which given to an amplification efficiency rate of 82. 5% ( 443/537 ). Totally, 140 normal homozygous, 112 heterozygotes and 155 affected homozygous embryos were identified, while 36 embryos had uncertain result. The successful diagnostic rate was 75.8% (407/537). After 3 days in the translocation groups, the rate of normal and/or balanced translocations in biopsed embryos with ≥7 cells was 34. 4% (77/224), which was significantly higher than 19. 6% ( 19/97 ) of biopsed embryos with ＜ 7 cells. After 4 days, the compaction rate in normal/balanced embryos was 59.4% ( 57/96 ), which was significantly higher than 34. 2% ( 77/225 ) in imbalanced embryos significantly. Seventy-five embryos transferred in 37 cycles with translocations group led to clinical pregnancy rate of 27.0% (10/37), and 170 embryos transferred in 58 cycles with Thalassaemia got a clinical pregnancy rate of 43. 1% ( 25/58 ) . Conclusions PGD can provide management efficiently for both chromosome translocations and Thalassaemia. Translocations might have slightly negative impact on embryo development before implantation.

Objective To compare the diagnostic efficiency between blastomere preimplantation genetic diagnosis (PGD) and polar body PGD for chromosomal translocation carriers. Methods Group A had 8 cycles using whole painting probes for the first polar body diagnosis, while group B had 29 cycles using two subtelomeric probes and one centromeric probe for the blastomere diagnosis. Results The fertilization rate of group A was significantly lower than group B [66. 1% (72/109) vs 85.2% (304/357) , P < 0.05]. There was no significant difference in the successful biopsy rate between two groups. However, group A had a significantly higher loss rate during fixation and higher no signal rate after fluorescence in situ hybridization [ FISH, 9. 6% (12/104) vs 1.6% (4/252), 11.2% (10/89) vs 3.0% (7/233) ]. Totally, the diagnostic efficiency in group A (72. 5% ,79/109 ) was significantly lower than that in group B( 89. 8%, 230/256, P < 0. 05 ). Although both the clinical pregnancy rate( 3/7 ) and implantation rate( 22. 2% ,4/18 ) of group A were higher, the differences were not statistically significant ( P > 0.05 ). Conclusion Both methods can be used efficiently in the PGD for chromosomal translocation carriers. Blastomere PGD has a higher diagnostic rate.

【Objective】To perform preimplanation genetic diagnosis by using dual color fluorescent in-situ hybridization (FISH).【Methods】The chemical and mechanical division methods were used to perform embryo biopsy in 3 cases of Robertsonian translocation t (13q14q).Vysis LSI 13q14 and Tel Vysion 14q probes were used to detect the blastomeres biopsied from the IVF embryos of the patients.FISH analysis was performed to select normal or balanced karyotype embryos ,which then were transfered into the uterus.【Results】Total of 23 oocytes were retrieved in 3 treatment cycles.Fertilization rate was 79%.14 embryos were available for embryo biopsy.Among them,9 embryos were biopsied by chemical division method,with further cleavage rate of 67%;5 embryos were biopsied by mechanical division method,with further cleavage rate of 40%.Single embryo was diagnosed as normal karyotype or balanced respectively in 2 treatment cycles.Both of them were transfered into the uterus.One clinical normal on-going pregnancy was achieved,the diagnosis was confirmed by amniocyte karyotype analysis.【Conclusion】Preimplanation genetic diagnosis can be used to resolve the problem of fertility for Robertsonian Translocation Carriers.

Objective:To evaluate the therapeutic efficacy of interposition urethroplasty on glandular hypospadias and severe penile curvature.Methods:A total of 9 cases of congenital glandular hypospadias and 2 cases of congenital severe penile curvature (>30°) treated with the interposition urethroplasty technique in the Shenzhen Children′s Hospital from November 2008 to December 2019 were retrospectively analyzed.The mean age of initial surgery was 40 (25-109) months.Two cases were surgically treated with one-staged interposition urethroplasty and the remaining were treated with two-staged interposition urethroplasty.They were followed up for 8 months to 12 years, including the penile morphology, urination, urethrocutaneous fistula, urethral stricture and uroflowmetry.Continuous variables were presented as ± s deviation, Student′s t-test was used for comparison between groups. Results:The median length of interposition urethra was 3.2 (2.2-4.2) cm.The cosmetic appearance of penis was good without residual chordee, urethral stricture or urethral diverticulum in all patients.Two cases had urethrocutaneous fistula(one-staged repair and two-staged repair were performed in one case respectively), which were successfully repaired by re-operation.The maximum of uroflowmetry in patients at 3 months [(8.3±1.0) mL/s] and 6 months [(6.7±1.9) mL/s] after surgery was significantly lower than that of children in healthy control group [(10.5±3.7) mL/s] ( t=3.221, 3.864, all P<0.05). However, there was no significant difference in 1-year maximum of uroflowmetry postoperatively between surgically treated patients [(10.5±3.7) mL/s] and healthy control group ( P>0.05). Conclusions:Interposition urethroplasty is an effective and safe treatment for glandular hypospadias and/or severe penile curvature in children.