Objective To compare the effect of different cryoprotectants and different concentrations on controlled?rate freezing of human ovarian tissues. Methods Ovarian tissues were sampled from 15 patients undergoing benign ovarian tumor surgery. Cortical slices were frozen by controlled?rate freezing using three cryoprotectants,propanediol,ethanediol,and dimethylsulphoxide,and the concentration of each cryoprotectant was 1.5 mol/L or 2.0 mol/L. Cortical slices obtained from each patient were processed with each cryopreservation procedure simultaneously. Morphology of fol?licles was studied by light and electron microscopy and the normal rate was compared with that of the fresh tissues from the patient. Results There were no significant differences in distribution of follicles of different developmental stages between each group(P<0.05). Light microscopy showed 1.5 mol/L EG and 2.0 mol/L EG groups had the best freezing effect,and the difference in the morphologically normal rate of follicles was not statisti?cally significant compared to fresh controls(P>0.05). However,the difference was statistically significant for 1.5 mol/L PROH,2.0 mol/L PROH, 1.5 mol/L DMSO and 2.0 mol/L DMSO groups(P<0.05). Electron microscopy showed the oocyte quality declined after controlled?rate freezing pro?cedure. However,the statistical analysis was not conducted due to little data. The viability of granulosa cells was significantly declined after all the freezing procedures compared to that of the fresh control tissues(P<0.01). The number of morphologically normal granulosa cells was slightly higher in the tissues which had been cryopreserved with 2.0 mol/L PROH and 1.5 mol/L EG,but no significant differences were found between any of two frozen groups(P>0.05). Conclusion Controlled?rate freezing using 1.5 mol/L EG as the cryoprotectant can better save oocytes and granulosa cells. It is a preferable freezing procedure for ovarian tissues.

Objective To summarize experience and prognosis of repeat renal transplantation after graft loss due to BK virus nephropathy (BKVN).Methods The clinical data of 4 adult patients undergoing repeat transplantation after previous allograft loss due to BKVN were collected and analyzed retrospectively.Results Three of four patients had documented allograft loss caused by BKVN and underwent retransplantation 5 months,9 months and 9 months respectively after hemodialysis with confirmed clearance of viremia.Allograft nephrectomy was performed on 1 of 3 patients 4 months before retransplantation.Maintenance immunosuppression was CsA + MMF + Pred,Tac + MMF + Pred and CsA + Pred in these 3 patients respectively.During the follow-up period of 9 months,5 months and 26 months,viremia kept negative and allografts function stabled normally without recurrence of BKVN.The cause of allograft loss was not illustrated in the other patient before retransplantation,which was performed without dialysis or allograft nephrectomy.BK virus was not monitored routinely after the operation.Four months later,his serum creatinine rose up to 400μmol/L and BKVN recurrence was proved by pathological analysis of the biopsy samples of the first and the second transplantation.Tac was switched to CsA and his serum creatinine declined to 260 μnol/L at 20th month.Conclusion Retransplantation can be performed on the patients with previous allograft loss due to BKVN.Allograft nephrectomy,clearance of viremia,monitoring BK virus and timely adjustment of immunosuppression were the keys to guarantee successful retransplantation.

Objective: To investigate the clinical manifestation, cytogenetics, gene mutations and prognostic factors of chronic neutrophilic leukemia (CNL) . Methods: 16 CNL cases, according to WHO (2016) -definition, were reviewed retrospectively. Identifications of the CSF3R, ASXL1, SETBP1, CALR and MPL mutations were performed by direct sequencing. JAK2 V617F mutation was detected by AS-PCR. Results: Of the 16 CNL patients, the median age was 64 (43-80) years with a male predominance of 75% (12/16) . The median hemoglobin was 114 (81-154) g/L, with median WBC of 41.20 (26.05-167.70) (10⁹/L and median PLT of 238 (91-394) ×10⁹/L.The median level of marrow fibrosis (MF) was 1 (0-3) degree. There was no other cytogenetic abnormalities except t (1;7) (p32;q11) , +21 and 14ps+ for each. All the 16 CNL patients harbored CSF3R T618I mutation. ASXL1 mutations were identified in 81% (13/16) , while SETBP1 mutations were confirmed in 63% (10/16) . The CALR K385fs*47 mutation was found. There was no mutation in JAK2 V617F or MPL in the above 16 patients. The median overall survival (OS) of patients presented with WBC≥50×10⁹/L at diagnosis (11 months) was significantly shorter than of WBC<50×10⁹/L (39 months, P=0.005) . Conclusion CSF3R T618I mutation was specific for CNL. The median OS of CNL patients was 24 months, and WBC≥50×10⁹/L at diagnosis was an unfavorable prognostic factor.