Objective To study the apoptosis of multiple myeloma cell line KM-3 induced by NK cells. Methods WST-1 assay was used to detect the killing effect of KM-3 cells treated with NK cells at different effector(E):target(T) ratio. Flow cytometry was applied to analyze Annexin-V+/PI- apoptotic cells and the mitochondrial transmembrane potential. Results NK cells could significantly kill KM-3 cells in a dosand time-dependent manner (P <0.05). After KM-3 cells- were treated with NK cells for 48 hours, the Annexin-V+/PI- cells were increased obviously in dose-dependence (P <0.05). The Annexin-V+PI- cells were increased in time-dependence when treated with NK cells(E:T ratio at 10:1) (P<0.05). The mitochondrial transmembrane potential of KM-3 cells treated with NK cells were significantly decreased in dose-and time-dependence (P < 0.05). Conclusion NK cell can kill KM-3 cells and induce apoptosis in a dose-and time-dependence manner.

Objective To establish a new methA of detecting vWF function. Methas The capability of vWF to bind collagen was evaluated with ELISA. Results The assay′s sensitivity was 0.001 U/ml. Coefficient of variation for inner-batch and inter-batch were 3.34 and 6.70 respectively.The vWF:CBA value of plasma was(90.24?22.87)% in 20 normal subjects. The vWF:CBA value was (31.94?27.36)% in 54 vWD, (35.22?20.02)% in 10 type 1 vWD, (8.74?6.38)% in 10 type 2A vWD and (0.70?0.58)% in 6 type 3 vWD,the values of all four vWD groups were lower than that of normal group( P

Objective To investigate mRNA and protein expression of aproliferation-inducing ligand (APRIL) in peripheral blood mononuclear cell and plasma of patients with B cells non-Hodgkin lymphoma (B-NHL) pre- or post- chemotherapy and to explore the role of APRIL in the B-NHL. Methods The mRNA and protein expression of APRIL were detected by real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) and enzyme-linked immunosorbent assay(ELISA), respectively. According to the standard curves,the quantitative levels of target mRNA and protein were determined. Results The detection linear range of targeted mRNA by RFQ-PCR was 101-109 pg/ml, and the coefficient of variation values for both intra- and inter-experimental reproducibility ranged 1.69 %-5.99 % and 6.35 %-10.12 %, respectively. To detect expression of APRIL protein by ELISA, the correlation coefficient of standard curves reached 0.9922. Before or after chemical treatment, the expression levels of APRIL mRNA and protein in patients with B-NHL were significantly higher than those in normal control (P ＜0.01), while the expression levels in post-treatment patients with Ⅲ and Ⅳ stage were lower than those of pre-treatment patients with corresponding stage (P ＜0.05, respectively), but those of pre- and post-treatment patients with Ⅰ / Ⅱ stage were not different (P ＞0.05).Conclusion The expression level of APRIL mRNA is similar to that of APRIL protein. APRIL may be involved in pathogenesis and development of B-NHL. Moreover, APRIL may be related to the burden of B-NHL and it may be considered as a targeted molecule for B-NHL.

Objective To explore the clinical diagnosis and treatment of hyperthyroidism patients coexisted thyroid carcinoma.Methods A retrospective analysis was made in 15 cases with hyperthyroidism patients coexisted thyroid carcinoma who were operated from Nov.2008 to Dec.2017 in Department of General Surgery of Jian'an District Peoples' Hospital.Results The incidence of hyperthyroidism patients coexisted thyroid carcinoma was 8.19％(15/183) in our study.All 15 patients included 12 papillary thyroid carcinoma,1 follicular thyroid carcinoma,1 medullary thyroid carcinoma,and 1 follicular papillary carcinoma.Among them,papillary thyroid microcarcinoma accounted for 66.67％(10/15).All 15 postoperative patients were followed up and the mean time was 13.25 months.Neither recurrence nor mortality occurred during the period.Conclusions Hyperthyroidism patients coexisted thyroid carcinoma have no characteristic clinical manifestations or specific diagnosis indicators.The prognosis can be good through strengthening awareness,improving vigilance,comprehensive analysis and surgical treatment.

Objective: To investigate the effect of minimal residual disease (MRD) monitoring by multiparameter flow cytometry (MFC) pre-conditioning on prognosis of acute myeloid leukemia in first complete remission (CR₁-AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) , and to explore the value of MRD monitoring by MFC in the prognosis evaluation on allo-HSCT in CR₁-AML. Methods: Between April 2012 and March 2015, consecutive 186 patients with CR₁-AML who underwent allo-HSCT were analyzed retrospectively. MRD in BM before conditioning was detected by eight-color MFC. Any level of residual disease was considered to be MRD positive. Results: ①Of 186 patients, MRD was negative in 151 patients, positive in 35 patients (<1% in 25 patients and 1% to 3% in 10 patients) . ② With the median follow up of 18 (5-41) months, two-year DFS was 80.0% (95%CI 68.5%-92.3%) . Univariate analysis showed that MRD positive patients had lower DFS[62.9% (95%CI 50.6%-75.2%) vs 88.9% (95%CI 76.6%-100.0%) , P<0.001], higher relapse[11.4% (95%CI 4.1%-29.0%) vs 3.3% (95% CI 0.6%-20.9%) , P=0.003] and higher NRM [25.7% (95% CI 8.1%-43.3%) vs 7.9% (95% CI 1.3%-26.5%) , P=0.001] after HSCT compared with that of MRD negative patients. Secondary AML showed lower DFS than primary AML [60.0% (95% CI 42.4%-76.6%) vs 86.0% (95% CI 68.4%-100.0%) , P=0.004]. ③Multivariate analysis indicated that MRD positive pre-HSCT was the independent risk factor on DFS [HR=4.565 (95%CI 2.918-9.482) , P<0.001], relapse [HR=5.854 (95%CI 1.538-22.288) , P=0.010] and NRM [HR=3.379 (95%CI 1.361-8.391) , P=0.009] after allo-HSCT in CR₁-AML. Conclusion MRD positive pre-conditioning was the only negative impact factor for patients with CR₁-AML after allo-HSCT. MRD by MFC can be used to assess the prognosis of CR₁-AML after allo-HSCT.

Objective: To analyze the clinical characteristics and prognosis of 34 cases of acute myeloid leukemia (AML) with FLT3 internal tandem duplication (FLT3-ITD) and MLL gene rearrangement. Methods: The clinical data of 34 AML patients with FLT3-ITD and MLL gene rearrangement was compared and analyzed for the therapeutic efficacy, prognostic factors when treated with chemotherapy, chemotherapy combined with targeted therapy or allogenic hematopoietic stem cell transplantation (allo-HSCT). Results: Of the thirty-four cases with median age 41 (4-71) years old, 63.6% presented with white blood cells (WBC) greater than 30×10⁹/L, 39.4% greater than 50 × 10⁹/L respectively on admission. M₅ (35.3%) made up the highest proportion. The cytogenetic abnormality reached 61.8%, of which the complex cytogenetic abnormality accounted for 11.8%. Eleven patients (32.35%) had both FLT3-ITD and MLL gene abnormalities. In addition to FLT3 and MLL abnormalities, 23 patients (67.6%) had one or more other gene abnormalities (multiple gene abnormalities). Of the 34 cases, 29.4% patients went into complete remission (CR) after two courses of chemotherapy. 20.6% (7 patients) went into CR after 3 or more courses of chemotherapy. The rate of early relapse in the CR group was 52.9%. Patients with WBC>50×10⁹/L or multiple gene abnormalities had a lower remission rate (7.7%, 5.4%) after two courses of chemotherapy. CR rate for the patients with more than three gene abnormalities was 0. The total 2-year overall survival (OS) in the 34 patients was 28.8% (95% CI 13.5%-46.0%) and the disease-free survival (DFS) was 27.1% (95% CI 12.5%-44.0%). Of the 18 patients treated with chemotherapy alone or chemotherapy combined with targeted therapy, 17 cases died within 2 years and 1 lost follow-up after giving up treatment. For the 16 patients received allo-HSCT, the 3-year OS was 43.4% (95% CI 13.7%-70.4%) and DFS 42.7% (95% CI 13.4%-69.7%). Conclusion AML patients with FLT3-ITD and MLL gene rearrangement often presented with M₅, accompanied by hyperleukocytosis, cytogenetic or multiple gene abnormalities. Those patients were observed to have low response rate and high early relapse when treated with chemotherapy without allo-HSCT. Patients had multiple gene abnormalities may be an important poor prognostic factor. Allo-HSCT is an effective treatment which could significantly improve the prognosis and survival of AML patients with FLT3-ITD and MLL gene abnormalities.

OBJECTIVETo evaluate the efficacy of HLA- haploidentical donor hematopoietic transplantation (Haplo- HSCT)for severe aplastic anemia (SAA)by compared with the same period of unrelated donor transplantation (UD- HSCT).

METHODSOf a cohort of 50 SAA patients between September 2012 and July 2014, 26 patients underwent UD- HSCT and 24 patients Haplo- HSCT.

RESULTSOS rate was 91.3% with a median follow-up of 9 (2-26)months. According to transplant type, there was no significant difference between UD- and Haplo-HSCT (96.1%vs 86.0%,P=0.30). 3 of 50 (6%)patients had primary engraft failure. Haplo- HSCT developed higher significantly incidence of Ⅱ- Ⅳ aGVHD (37.5%vs 3.83%,P=0.003)and cGVHD (37.5%vs 15.3%,P=0.030)than UD-HSCT. Haplo-HSCT also had significantly higher incidences of CMV viremia (78.2%vs 46.1%,P=0.005)and EBV viremia (43.1%vs 16.0%,P=0.040), respectively than UD-HSCT. But the incidences of hemorrhagic cystitis were similar between two transplant types (39.1%vs 23.0%,P=0.120).

CONCLUSIONThis study showed favorable outcome of Haplo-HSCT for SAA, which was comparable with UD-HSCT.

Objective: To investigate three different types of donor hematopoietic stem cell transplantation (HSCT) for intermediate and high-risk myelodysplastic syndrome (MDS) . Methods: Between August 2001 and May 2015, 167 consecutive patients with MDS in intermediate and high-risk who underwent allogeneic HSCT were analyzed retrospectively. Results: With the median follow up of 60 (12-177) months, The total 5-year DFS was 67.8% (95%CI 60.0%-75.6%) . Among three different types of donor, 5-year DFS rates were 68.0% (95%CI 54.1%-81.9%) in MSD-HSCT vs 77.4% (95%CI 62.1%-92.7%) in MUD-HSCT vs 64.0% (95% CI 52.4%-75.6%) in Haplo-HSCT (P=0.632) , respectively. Univariate analysis showed that median disease course before HSCT was the influencing factor of DFS (P=0.018) . Five-year relapse and TRM had no correlation with the above-mentioned factor. Conclusions Haplo-HSCT for intermediate and high-risk MDS achieved similar effect produced by MUD or MSD, Haplo-HSCT could be used as an important alternative donor. allo-HSCT must be performed on intermediate and high-risk MDS patients as early as possible after diagnosis.

Objective: To analyze the effect of NCCN (2015) risk stratification on prognosis of patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Retrospective analysis of 258 patients with AML in CR (186 cases in CR₁, 72 cases in CR₂) who underwent allogeneic HSCT in our hospital between April 2012 and March 2015 according to NCCN (2015) risk stratification. Of them, 63 cases were classified as low risk, 112 cases intermediate risk and 83 cases high risk. Results: ①With the median follow up of 18 (5-41) months, two-year disease free surviva (DFS) in 258 patients was 78.0% (95% CI 60.4%-96.6%) . Two-year DFS in AML after transplantation was 78.6% (95% CI 61.0%-96.2%) in low risk, 76.0% (95% CI 84.0%-93.6%) in intermediate risk and 80.3% (95% CI 62.7%-97.9%) (P=0.886) in high risk groups respectively. ②Univariate analysis showed that DFS has no significant difference in patient age, the median disease course before HSCT, the WBC number at the beginning of the disease, blood routine and chromosomes examination before transplantation, extramedullary disease before transplantation, disease status before transplantation, conditioning regimen, donor type, donor and recipient sex, recipient blood type, transfused MNC number, transfused CD34⁺ cell number and transfused CD3⁺ cell number. DFS was significant lower in primary AML than that in secondary AML (P=0.006) and also lower in MRD positive than that in MRD negative (P=0.003) . The accumulative relapse was significant higher in CR₂ compared to that in CR₁ (P=0.046) . Accumulative non-relapse mortality (NRM) was significanlyt higher in secondary AML compared to that in primary AML (P=0.004) and also higher in MRD positive compared to that in MRD negative (P=0.010) . ③Multivariate analysis showed that MRD positive was the only significant factor in DFS and NRM. Conclusion Allo-HSCT treatment of AML CR patients could achieve a high efficacy, which is similar between CR₁ and CR₂ patients. There is no significant correlation between NCCN (2015) risk stratification and the prognosis of AML patients with allo-HSCT treatment. Pre-conditioning MRD status monitored by multiparameter flow cytometry was the only impact factor on DFS and NRM in allo-HSCT for CR-AML patients.