Cord blood is the main source of stem cell outside of bone marrow and peripheral blood, and cord blood transplantation, especially unrelated cord blood transplantation (UCBT) has been widely used. The major challenges in UCBT are delayed engraftment and graft rejection. Many approaches have been investigated to enhance collection of HSC and HPC in cord blood units,sinduding use of double unit UCBT, adopt reduced intensity conditioning (RIC) approach, amplification of cord blood cells in vivo or ex vivo, and co-infusion with a haploidentical T cell depleted graft or mesenchymal stem cells(MSC),or injecting cord blood cells directly into the bone marrow.Those approaches may greatly increase the clinical use of cord blood cells for transplantation.

Objective To clone human beta-2 adrenoceptor gene from human bladder smooth muscle and to construct its antisense eukaryotic expression vector. Methods The ?_2-AR full length cDNA was cloned from human detrusor cells through RT-PCR and subcloned into clone vector (pUC18).The objective gene was then cut from ClaⅠ/HindⅢ sites of pUC18 with restriction endonulcease and subcloned into pLNCX vector in trans-direction.Finally the constructed ?_2-AR gene antisense expresstion vector was identified through restriction endonuclease analysis and sequencing. Results The sequence of cloned ?_2-AR full length cDNA was certified by comparison with the database of the Genebank.The constructed antisense eukaryotic expression vector was proved to be same with designed by restriction endonuclease analysis and sequencing. Conclusions ?_2-AR full length cDNA was cloned and its antisense eukaryotic expression vector was successfully constructed.This technique establishs the foundation for the further research on drug treatment of bladder dysfunction.

Objective To investigate the efficacy and safety of posaconazole in the prophylaxis or salvage treatment of invasive fungal disease (IFD) in patients with hematological diseases.Methods 25 patients with hematological diseases receiving posaconazole treatment from Feb 2014 to Feb 2015 were analyzed retrospectively.The patients' average age was 32.6 years old (16-64 years old).18 patients received posaconazole for IFD prophylaxis, and 7 patients for IFD salvage treatment.Results 18 patients receiving posaconazole for IFD prophylaxis had no clinical manifestation of IFD (that is no case of breakthrough fungal infection) during treatment or in the 12 weeks after treatment, and the average prophylaxis period was 21 d (14-35 d).Among 7 patients receiving posaconazole for IFD salvage treatment, 6 patients were effective, including 4 cases cured and 2 cases effective.There were no obviously side effects of posaconazole in these patients.Conclusion Posaconazole has good clinical response for IFD in the hematologic diseases patients whether in prophylaxis or in salvage treatment.

Objective:To explore the clinical characteristics, diagnosis and treatment of acute leukemia patients during pregnancy.Methods:The clinical data of 16 cases with acute leukemia during pregnancy from January 2009 to December 2018 in the First Affiliated Hospital of USTC were retrospectively analyzed. The diagnosis and treatment regimens, pregnancy outcome, the early fetus and survival status of patients were also analyzed.Results:All 16 leukemia cases were confirmedly diagnosed and classified by bone marrow puncture, including 13 cases of acute myeloid leukemia (5 cases of non-acute promyelocytic leukemia and 8 cases of acute promyelocytic leukemia) and 3 cases of acute lymphoblastic leukemia. At the time of confirmed diagnosis, 6 patients were in first trimester, 6 cases in second trimester and 4 cases in late trimester. As for pregnancy outcome, 1 patient had natural birthing, 5 patients underwent cesarean operation, 9 patients underwent artificial abortion and 1 patient had spontaneous abortion. Chemotherapy was performed in 15 patients during pregnancy, 11 patients received chemotherapy for treatment of primary disease after pregnancy, 3 patients died during the treatment. During the follow-up of 13 cases, 8 patients survived and 5 patients lost follow-up.Conclusions:Early diagnosis of acute leukemia during pregnancy is very important. Bone marrow puncture should be performed timely to make clear diagnosis when blood routine result is abnormal during antenatal care. Multidisciplinary consultation should be initiated in time, and the best treatment plan should be worked out to guard against serious complications during pregnancy.

Objective To set up a real-time quantitative PCR approach for detection and quantification for bcr-abl transcripts in CML patients,and detect minimal residual disease (MRD) in CML by real-time quantitative PCR (RQ-PCR)and evaluate the significance of MRD detection.Methods The ber-abl.fusion gene expression in 80 patients with CML was analyzed by RQ-PCR. The patients were divided into three groups according to the different treatment, allogeneic hematopoietic stem cell transplantation group,imatinib group and hydroxyurea group. The change of bcr-abl fusion gene was monitored in CML patients before and after treatment.Results The average of RQ-PCR detection on newly diagnosed patients with CML in chronic phase was 6847.67 copies / 104 cells,the accelerated phase was 306 176.08 copies / 104 cells,and the average results were 944.33, 2.37, 0.29, 0 copies / 104 cells after allogeneic hematopoietic stem cell transplantation one month,6 months,12 months or 24 months respectively.The average of RQ-PCR detection after use imatinib mesylate 3 months was 3720.23 copies / 104 cells and not be detected after one year. The average was 7290.11 and 3143.24 copies / 104 cells after hydroxyurea treatment 0 and 9 months respectively.The difference in first two groups was not significant (t=1.74,P=0.17), but the difference between the third group and the first two groups was significant (t=3.74,P=0.01.t=2.97,P=0.02). The upregulation of bcr-abl transcript levels could be detected when disease progression. The transcripts level in accelerated phase was significantly higher than that in chronic phase. Conclusion RQ-PCR can be used to detect the MRD,monitor the treatment outcome,predict disease recurrence and give early intervention.

ObjectiveTo explore the genetic abnormalities of multiple myeloma (MM)patients by fluorescence in situ hybridization (FISH).MethodsWith the application of FISH,sequence specific DNA probes (IGH,DI3S319/p53 and 1q21/RB1) were applied to detect 14q32 rearrangement,del(13q14),del (17p13)and gain of lq21.Forty-four MM patients were enrolled in this study.ResultsThirty-two cases (72.7 ％) detected by FISH had genetic abnormality in 44 cases,lq21 amplification was observed in 11 cases (25.0 ％),while RB1 deletion in 17 cases (38.6 ％),D13S319 deletion in 16 cases (36.4 ％),p53 deletion in 6 cases(13.6 ％)and 14q32 translocation in 19 cases(43.2 ％).The patients with one abnormality was detected in 10 cases(22.7 ％),two abnormalities in 11 cases(25.0 ％),three abnormalities in 8 cases (18.2 ％),4 abnormalities in 3 cases(6.8 ％).28 were found to undergo split-phase by conventional cytogenetic in 44 patients.The patients with genetic abnormalities detected by conventional G-banding was 2 cases (7.14 ％),the difference with that in FISH was significant (P ＜0.05).Genetic abnormalities compared with clinical parameters showed that β2-MG in IGH gene abnormal patients were significantly higher than those without such abnormalities (P ＜0.05).Patients with bone marrow plasma cells of lq21 amplification were higher than those with normal karotypes(P ＜0.05),CRE was significantly higher among lq21 amplification and p53 deletion patients (P ＜0.05),CRP was significantly higher among p53 deletion patients (P ＜0.05).No significant difference was oberserved in relationship of the chromosome aberration and age,the chromosome aberration and stage.ConclusionThe most common genetic abnormalities in MM is IGH rearrangement and absence of RB1 and D13S319,followed by lq21 amplification,the least is p53 deletion.FISH is a rapid and sensitive technique to refine chromosome aberrations in MM.The specific detection for genomic features of MM is proved to be correlative with its clinicopathologic characteristics and the prognosis.

Objective To study the curative efficacy and safety of single-unit umbilical cord blood transplantation (sUCBT) for malignant hematologic diseases,which is provided by China's public cord blood bank.Methods We retrospectively analyzed 409 cases of malignant hematologic diseases who accepted myeloablative single-unit unrelated donor UCBT without ATG at our center between May 2008 and December 2016.A comparative analysis was made on the total nuclear cells (TNC) of the umbilical cord blood before freezing and after thawing,the cells of CD34+,the recovery rate of cells and the clinical effect of UCBT.Result 409 units of umbilical cord blood used in UCBT respectively came from eight China's public cord blood banks.The average TNC of 409 units of umbilical cord blood before freezing and after the tubular recovery were respectively 18.5 × 108 and 16.34 × 108 (p =0.000).The average recovery rate of the tubular recovery was 88.5％,and there was significant difference among cord blood banks (P =0.000).The average TNC of umbilical cord blood before freezing and transfusion were respectively 18.5 × 108 and 15.86 × 108 (p =0.000).The average recovery rate of umbilical cord blood transfusion was 85.9％,with the difference being significant among cord blood banks (P =0.000).The average number of CD34+ cells before freezing and after the tubular recovery was 11.18 × 106and 8.68 × 106 (p =0.000).The average recovery rate of CD34+ cells after the tubular recovery was 80.75 ％,with the difference being significant among the cord blood banks (P =0.000).At 42nd day after UCBT,the cumulative incidence of neutrophil engraftment was 95.4％,and the median time of the engraftment was 17 days (11-38 days).The cumulative incidence of platelet engraftment at 120th day was 84.6％,and the median time of the engraftment was 36 days (14-93 days).The cumulative incidence of erythrocyte engraftment at 60th day was 92％,and the median time of engraftment was 22 days (9d-60 days).After the umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of engraftment.The P values for cumulative incidence of neutrophil,platelet and erythrocyte engraftment were respectively 0.004,0.01 and 0.000 2,with the differences being statistically significant.At 100th day after UCBT,the cumulative incidence of Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees of acute graft-versus-host disease (aGVHD) was respectively 28.63％ and 15.7％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of aGVHD.There was no significant difference between Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees (P =0.809 and 0.68 respectively).At 3rd year after UCBT,the cumulative incidence of relapse was 15.89％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the cumulative incidence of relapse (P =0.898).At 3rd year after UCBT,the overall survival (OS) rate and disease free survival (DFS) rate were respectively 66.7％ and 59％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in OS and DFS.There was no significant difference in OS and DFS (P =0.566 and 0.703 respectively).At 3rd year after sUCBT,the rate of graft-versus-host diseases/relapse-free survival (GRFS) was 54.3％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the rate of GRFS (P =0.449).Conclusion The umbilical cord blood provided by China's public cord blood bank was used in UCBT.It has a high safety and good efficacy in treating malignant hematologic diseases.But it needs to set up the standardized and normalized quality-control system of umbilical cord blood for China's public cord blood bank.

Objective To analyze the epidemiologic and clinical features of post-engraftment blood stream infection (pePSI) after unrelated cord blood transplant (UCBT) in our hospital,and provide the basis for empiric antibacterial treatment.Methods 484 patients with hematological malignancies who received single-unit high intensity myeloablative UCBT in our hospital between April 2011 and November 2017 were enrolled.The incidence,etiology of BSI and associated mortality,drug resistance rate in the post-engraftment phase were investigated.Results Totally 25 episodes of BSI among 22 patients in the post-engraftment phase were documented,and the incidence of peBSI was 5 ％.Gram-negative organisms predominated over Gram positive,with Escherichia coli being the most frequent Gram-negative organism isolated (31.5％).Among Gram positive organisms,methicillin resistant Staphylococcus (MRS) was the most frequent species isolated (66％).Nearly 33％ of Escherichia coli isolates and 60％ Klebsiella pneumonia isolates were carbapenem-resistant.All Grampositive bacteria were sensitive to vaneomyein and linezolid.Among the 22 patients,14 patients were cured and survived (63％) eventually.Conclusion The most frequent causative agents of the peBSI after UCBT were Escherichia coli,Klebsiella pneumonia and MRS,etc.Combined antibacterial treatment including a carbapenem or beta lactamase inhibitor can be used for patients suffering fever in the post-engraftment phase as empiric antibacterial therapy.Vaneomyein and linezolid can be used as the first-line therapy for Gram-positive bacteria.

Objective: To compare the efficacy of unrelated cord blood transplantation (UCBT) and HLA-identical sibling peripheral blood stem cell transplantation (PBSCT) for the treatment of adult hematological malignancies. Methods: From April 2011 to December 2015, a total of 81 patients receiving single-unit UCBT and 57 patients receiving HLA-identical sibling PBSCT were enrolled in this study. All of the patients received myelablative conditioning. Cyclosporine combined with mycophenolate mofetil was adopted for GVHD prophylaxis. Results: The cumulative incidence of neutropil engraftment at day-42 was 95.0% and 100% in UCBT and sibling PBSCT groups, respectively (P=0.863) . Platelet engraftment at day 100 was 87.3% (95%CI 76.8%-93.1%) in UCBT group, which was significantly lower than that of sibling PBSCT group[98.2% (95%CI 87.3%-99.7%) ] (P=0.005) . There were no significant differences in terms of Ⅱ-Ⅳ acute GVHD or Ⅲ-Ⅳ acute GVHD in two groups (P=0.142, 0.521) . The 3-year chronic GVHD and extensive chronic GVHD were 14.9% (95%CI 5.2%-23.5%) and 11.2% (95%CI 2.9%-18.7%) , respectively in UCBT group, which was significantly lower than that of sibling PBSCT group[35.2% (95%CI 19.4%-47.8%) , 31.4% (95%CI 16.2%-43.9%) ] (P=0.008, 0.009) . The 3-year transplant-related mortality (TRM) was similar between two groups (30.1% vs 23.2%, P=0.464) . The relapse rate at 3-year in UCBT group[12.9% (95%CI 6.6%-21.5%) ]was significantly lower than that in sibling PBSCT group[24.3% (95%CI 13.5%-36.8%) ] (P=0.039) . There were no significant differences in terms of overall survival (OS) and disease-free survival (DFS) between two groups (58.6% vs 54.8%, P=0.634; 57.0% vs 52.4%, P=0.563) . But GVHD-free and relapse-free survival (GRFS) in UCBT group [55.7% (95%CI 44.1%-65.8%) ]was significantly higher than that of sibling PBSCT group[42.9% (95%CI 29.8%-55.3%) ] (P=0.047) . Conclusions For adult hematological malignancies, the incidences of acute GVHD and TRM were similar between UCBT and sibling PBSCT recipients, and the incidences of chronic GVHD and relapse were lower in UCBT recipients. UCBT recipients had higher GRFS rate although OS and DFS were similar between two groups, which may reflect the real recovery and better quality of life following UCBT.

Objective: To explore the clinical efficacy and safety of unrelated umbilical cord blood transplantation (UCBT) in the treatment of refractory and relapsed acute leukemia (AL) patients. Methods: The clinical data of 22 refractory and relapsed AL patients who were treated with UCBT as salvage therapy from November 2009 to May 2017 were retrospectively analyzed. All patients received a myeloablative conditioning regimen for prevention of graft-versus-host disease (GVHD) with cyclosporine A (CSA)/short course of mycophenolate mofetil (MMF). Results: ①Of 22 patients, 9 cases were male and 13 female. The median age was 23 (15-44) years and median weight of 52.5 (43-82) kg. All patients were transplanted with a median umbilical cord blood nucleated cells of 3.07 (1.71-5.30)×10⁷/kg (by weight), the median CD34⁺ cells was 1.60 (0.63-3.04)×10⁵/kg (by weight). ②The myeloid cumulative implantation rate was 95.5% (95%CI 45.2-99.7%) after transplantation of 42 d, with the median implantation time of 19 (13-27) d. The platelet cumulative implantation rate after transplantation of 120 d was 81.8% (95%CI 54.2-93.6%), the median implantation time of 42 (20-164) d. ③The incidence of Ⅱ-Ⅳ, Ⅲ-Ⅳ aGVHD and the 2 year cumulative incidence of cGVHD were 36.4%, 13.6% and 40.3% respectively. ④ The transplant related mortality (TRM) after transplantation of 180d was 22.7%, 2 year cumulative rate of relapse was 18.7% (95%CI 3.6-42.5%), 2 year disease-free survival rate (DFS) and overall survival rate (OS) were 53.7% and 58.1%, respectively. Conclusion The preliminary results show that the use of UCBT is safe and effective for refractory and relapsed AL patients who fail to respond to conventional chemotherapy.