Comparison of the characteristics of NK cells after two different methods of expansion and observation of the clinical efficacy in patients who relapsed post allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

Objective: To explore the differences in the biological effects of different expansion systems on natural killer (NK) cells, as well as the safety and preliminary clinical efficacy in the treatment of patients with recurrence after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Peripheral blood cells from healthy donors were stimulated with either CD3 combined with CD52 or K562 feeder cells loaded with IL-21/4-1BB to induce NK cell expansion. Changes in the NK cell phenotype, cytokine secretion, and cytotoxicity before and after expansion were detected. We also evaluated the safety and clinical efficacy of two different expansion strategies for patients received NK infusion. Results: Compared with the CD3/CD52 monoclonal antibody amplification system, the feeder cell expansion group had a higher purity of NK cells and higher expression ratios of NK cell surface activation receptors such as DNAM-1 and NKp30, while inhibitory receptor CTLA-4 expression was low and NKG2D/CD25/CD69/ Trail/PD-1/TIM-3/TIGIT had no statistically significant differences between the groups. Further functional results showed that the expression level of KI67 in NK cells after expansion in the two groups increased significantly, especially in the feeder cell expansion group. Simultaneously, the perforin and granzyme B levels of NK cells in the feeder cell expansion group were significantly higher than in the CD3/CD52 expansion group. A retrospective analysis of eight patients who received monoclonal antibody-expanded NK cell reinfusion and nine patients with trophoblast cell-expanded NK cell reinfusion was done. The disease characteristics of the two groups were comparable, NK cell reinfusion was safe, and there were no obvious adverse reactions. Clinical prognostic results showed that in the CD3/CD52 monoclonal antibody amplification group, the MRD conversion rate was 50% (2/4) , and the feeder cell expansion group was 50% (3/6) . After 5 years of follow-up from allo-HSCT, three patients in the monoclonal antibody expansion group had long-term survival without leukemia, and the remaining five patients had died; two patients died in the feeder cell expansion group, and the other six patients had long-term survival. Six cases had GVHD before NK cell reinfusion, and GVHD did not aggravate or even relieved after NK cell reinfusion. Conclusions: Preliminary results show that the biological characteristics of NK cells with diverse expansion strategies are significantly different, which may affect the clinical prognosis of patients with recurrence or persistent minimal residual disease after HSCT. The two groups of patients treated with NK cells from different expansion strategies had no obvious adverse reactions after NK cell infusion, but efficacy still needs to be further confirmed.

A retrospective comparative study of haplotype hematopoietic stem cell transplantation and human leukocyte antigen-matched sibling donor hematopoietic stem cell transplantation in the treatment of acute B-lymphocyte leukemia / 中华血液学杂志

Objective: To investigate whether haplotype hematopoietic stem cell transplantation (haplo-HSCT) is effective in the treatment of pre transplant minimal residual disease (Pre-MRD) positive acute B lymphoblastic leukemia (B-ALL) compared with HLA- matched sibling donor transplantation (MSDT) . Methods: A total of 998 patients with B-ALL in complete remission pre-HSCT who either received haplo-HSCT (n=788) or underwent MSDT (n=210) were retrospectively analyzed. The pre-transplantation leukemia burden was evaluated according to Pre-MRD determinedusing multiparameter flow cytometry (MFC) . Results: Of these patients, 997 (99.9% ) achieved sustained, full donor chimerism. The 100-day cumulative incidences of neutrophil engraftment, platelet engraftment, and grades Ⅱ-Ⅳ acute graft-versus-host disease (GVHD) were 99.9% (997/998) , 95.3% (951/998) , and 26.6% (95% CI 23.8% -29.4% ) , respectively. The 3-year cumulative incidence of total chronic GVHD was 49.1% (95% CI 45.7% -52.4% ) . The 3-year cumulative incidence of relapse (CIR) and non-relapse mortality (NRM) of the 998 cases were 17.3% (95% CI 15.0% -19.7% ) and 13.8% (95% CI 11.6% -16.0% ) , respectively. The 3-year probabilities of leukemia-free survival (LFS) and overall survival (OS) were 69.1% (95% CI 66.1% -72.1% ) and 73.0% (95% CI 70.2% -75.8% ) , respectively. In the total patient group, cases with positive Pre-MRD (n=282) experienced significantly higher CIR than that of subjects with negative Pre-MRD [n=716, 31.6% (95% CI 25.8% -37.5% ) vs 14.3% (95% CI 11.4% -17.2% ) , P<0.001]. For patients in the positive Pre-MRD subgroup, cases treated with haplo-HSCT (n=219) had a lower 3-year CIR than that of cases who underwent MSDT [n=63, 27.2% (95% CI 21.0% -33.4% ) vs 47.0% (95% CI 33.8% -60.2% ) , P=0.002]. The total 998 cases were classified as five subgroups, including cases with negative Pre-MRD group (n=716) , cases with Pre-MRD<0.01% group (n=46) , cases with Pre-MRD 0.01% -<0.1% group (n=117) , cases with Pre-MRD 0.1% -<1% group (n=87) , and cases with Pre-MRD≥1% group (n=32) . For subjects in the Pre-MRD<0.01% group, haplo-HSCT (n=40) had a lower CIR than that of MSDT [n=6, 10.0% (95% CI 0.4% -19.6% ) vs 32.3% (95% CI 0% -69.9% ) , P=0.017]. For patients in the Pre-MRD 0.01% -<0.1% group, haplo-HSCT (n=81) also had a lower 3-year CIR than that of MSDT [n=36, 20.4% (95% CI 10.4% -30.4% ) vs 47.0% (95% CI 29.2% -64.8% ) , P=0.004]. In the other three subgroups, the 3-year CIR was comparable between patients who underwent haplo-HSCT and those received MSDT. A subgroup analysis of patients with Pre-MRD<0.1% (n=163) was performed, the results showed that cases received haplo-HSCT (n=121) experienced lower 3-year CIR [16.0% (95% CI 9.4% -22.7% ) vs 40.5% (95% CI 25.2% -55.8% ) , P<0.001], better 3-year LFS [78.2% (95% CI 70.6% -85.8% ) vs 47.6% (95% CI 32.2% -63.0% ) , P<0.001] and OS [80.5% (95% CI 73.1% -87.9% ) vs 54.6% (95% CI 39.2% -70.0% ) , P<0.001] than those of MSDT (n=42) , but comparable in 3-year NRM [5.8% (95% CI 1.6% -10.0% ) vs 11.9% (95% CI 2.0% -21.8% ) , P=0.188]. Multivariate analysis showed that haplo-HSCT was associated with lower CIR (HR=0.248, 95% CI 0.131-0.472, P<0.001) , and superior LFS (HR=0.275, 95% CI 0.157-0.483, P<0.001) and OS (HR=0.286, 95% CI 0.159-0.513, P<0.001) . Conclusion: Haplo HSCT has a survival advantage over MSDT in the treatment of B-ALL patients with pre MRD<0.1% .

A risk score system for stratifying the risk of relapse in B cell acute lymphocytic leukemia patients after allogenic stem cell transplantation / 中华医学杂志(英文版)

BACKGROUND@#For patients with B cell acute lymphocytic leukemia (B-ALL) who underwent allogeneic stem cell transplantation (allo-SCT), many variables have been demonstrated to be associated with leukemia relapse. In this study, we attempted to establish a risk score system to predict transplant outcomes more precisely in patients with B-ALL after allo-SCT.@*METHODS@#A total of 477 patients with B-ALL who underwent allo-SCT at Peking University People's Hospital from December 2010 to December 2015 were enrolled in this retrospective study. We aimed to evaluate the factors associated with transplant outcomes after allo-SCT, and establish a risk score to identify patients with different probabilities of relapse. The univariate and multivariate analyses were performed with the Cox proportional hazards model with time-dependent variables.@*RESULTS@#All patients achieved neutrophil engraftment, and 95.4% of patients achieved platelet engraftment. The 5-year cumulative incidence of relapse (CIR), overall survival (OS), leukemia-free survival (LFS), and non-relapse mortality were 20.7%, 70.4%, 65.6%, and 13.9%, respectively. Multivariate analysis showed that patients with positive post-transplantation minimal residual disease (MRD), transplanted beyond the first complete remission (≥CR2), and without chronic graft-versus-host disease (cGVHD) had higher CIR (P  < 0.001, P = 0.004, and P  < 0.001, respectively) and worse LFS (P  < 0.001, P = 0.017, and P  < 0.001, respectively), and OS (P  < 0.001, P = 0.009, and P  < 0.001, respectively) than patients without MRD after transplantation, transplanted in CR1, and with cGVHD. A risk score for predicting relapse was formulated with the three above variables. The 5-year relapse rates were 6.3%, 16.6%, 55.9%, and 81.8% for patients with scores of 0, 1, 2, and 3 (P  < 0.001), respectively, while the 5-year LFS and OS values decreased with increasing risk score.@*CONCLUSION@#This new risk score system might stratify patients with different risks of relapse, which could guide treatment.

Association of Persistent Minimal Residual Disease with Poor Outcomes of Patients with Acute Myeloid Leukemia Undergoing Allogeneic Hematopoietic Stem Cell Transplantation / 中华医学杂志(英文版)

Background@#Several studies have shown that detection of minimal residual disease (MRD) in acute myeloid leukemia (AML) is an independent prognostic factor. This study aimed to evaluate the significance of dynamic MRD pretransplantation on outcome of AML patients receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*Methods@#We retrospectively analyzed 145 consecutive AML patients undergoing allo-HSCT in complete remission status between June 2013 and June 2016. MRD was determined with multiparameter flow cytometry after the first and second courses of chemotherapy and pre-HSCT.@*Results@#In matched sibling donor transplantation (MSDT) settings, patients with positive MRD had higher cumulative incidence of relapse (CIR) than those without MRD after the first (32.3 ± 9.7% vs. 7.7 ± 3.1%, χ = 3.661, P = 0.055) or second course of chemotherapy (57.1 ± 3.6% vs. 12.5 ± 2.7%, χ = 8.759, P = 0.003) or pre-HSCT (50.0 ± 9.7% vs. 23.0 ± 3.2%, χ = 5.547, P = 0.019). In haploidentical SCT (haplo-SCT) settings, the MRD status at those timepoints had no significant impact on clinical outcomes. However, patients with persistent positive MRD from chemotherapy to pre-HSCT had higher CIR than those without persistent positive MRD both in MSDT and haplo-SCT settings. Patients with persistent positive MRD underwent MSDT had the highest relapse incidence, followed by those with persistent positive MRD underwent haplo-SCT, those without persistent MRD underwent haplo-SCT, and those without persistent MRD underwent MSDT (66.7 ± 9.2% vs. 38.5 ± 6.0% vs. 18.8 ± 8.7% vs. 12.0 ± 1.0%, χ = 20.763, P < 0.001). Multivariate analysis showed that persistent positive MRD before transplantation was associated with higher CIR (hazard ratio [HR] = 1.69, 95% confidence interval [CI]: 1.200-2.382, P = 0.003), worse leukemia-free survival (HR = 1.812, 95% CI: 1.168-2.812, P = 0.008), and overall survival (HR = 2.354, 95% CI: 1.528-3.627, P < 0.001).@*Conclusion@#Our results suggest that persistent positive MRD before transplantation, rather than positive MRD at single timepoint, could predict poor outcome both in MSDT and haplo-SCT settings.

Allogeneic hematopoietic stem cell transplantation for chronic myelomonocytic leukemia: a report of 12 patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To retrospectively review the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for chronic myelomonocytic leukemia (CMML).</p><p><b>METHODS</b>The engraftment, graft versus host disease (GVHD), infection, relapse and survival of 12 CMML patients received allo-HSCT were observed. The clinical outcome of allo-HSCT for CMML was analyzed.</p><p><b>RESULTS</b>Twelve (7 males and 5 females) CMML patients with a median age of 39 years old received allo-HSCT including 7 from HLA-matched sibling and 5 from haploidentical related donors. All 12 patients achieved engraftment. The median time of neutrophil engraftment and platelet engraftment were 15 (11 - 20) days and 13 (11 - 18) days, respectively. 4 patients occurred acute GVHD, and 3 occurred chronic GVHD. After the median follow-up of 17.5 months (12 - 32 months), the overall survival, disease free survival and relapse rate were 66.7%, 66.7%, and 16.7%, respectively.</p><p><b>CONCLUSION</b>Allo-HSCT can improve the survival of patients with CMML, and is a effective therapy for CMML.</p>

The kinetics and prognosis of platelet reconstitution after unmanipulated haploidentical stem cell transplantation without in vitro T cell depletion / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the kinetics of platelet reconstitution and its prognostic significance in patients received unmanipulated haploidentical stem cell transplantation (Haplo-HSCT) without in vitro T cell depletion.</p><p><b>METHODS</b>A total of 291 patients received Haplo-HSCT without in vitro T cell depletion between January 2007 to December 2008 were retrospectively reviewed. They were categorized into 3 groups according to the platelet count on day 30, day 60 and day 90: (1) persistent thrombocytopenia (Group A) was defined as the platelet count never reached 50×10⁹/L on the three time points; (2) unstable thrombocytopenia (Group B): the platelet count recovered to a level of 50×10⁹/L by day 30 or 60 or 90, yet did not reach a level of more than 100×10⁹/L; (3) non-thrombocytopenia (Group C): the platelet count was higher than 100×10⁹/L on day 90. The kinetics of platelet reconstitution, overall survival (OS) and treatment related mortality (TRM) were compared between 3 groups.</p><p><b>RESULTS</b>Of the 291 consecutive patients, 288 cases engrafted successfully and 262 cases were platelet transfusion independent. The median intervals of neutrophil and platelet engraftment were 13 (9-29) days and 17 (7-180) days, respectively. The cumulative incidence of grade III-IV acute graft versus host disease (GVHD) on day 100 and chronic GVHD at 3 years were 14.7% and 56.4% respectively. OS and TRM at 3 years were 64.6% and 22.3% respectively. At the end of the follow-up, 266 cases were platelet transfusion independent: including 71 (24.4%) cases in Group A, 147 (50.5%) in Group B and 73 (25.1%) in Group C. OS in group A, B and C was 38.0%, 69.4% and 80.8% (P<0.05) respectively. TRM in Group A, B and C was 53.5%, 17.7% and 1.4% (P<0.05) respectively. Persistent thrombocytopenia was related with lower OS and higher TRM in multivariate analysis.</p><p><b>CONCLUSION</b>Persistent thrombocytopenia was common after Haplo-HSCT without in vitro T cell depletion, and patients with persistent thrombocytopenia have poor OS and higher TRM.</p>

The relationship between serum resistin level and blood sugar, blood lipids and thyroid hormone in patients with hyperthyroidism / 中国地方病学杂志

ObjectiveTo undertake a controlled, prospective study to investigate the relationship between serum resistin level and blood sugar, blood lipids and thyroid hormone in patients with hyperthyroidism. Methods Clinical data of 50 patients with hyperthyriodism were collected from 2008 to 2009 in department of endocrinology the Second Hospital of Harbin Medical University. All patients were newly diagnosed cases that received no medication. At the same time, 40 healthy persons underwent physical examination in the hospital were taken as control group. Diabetes, obesity, hypertension, and high cholesterol were excluded from the two groups. Serum resistin level was determined using ELISA method. Chemiluminescence was used to check the levels of fasting insulin, free triiodothyronine (FT3), free thyroxin (FT4), and thyroid stimulating hormone (TSH). Glucose oxidase method determination-peroxidase-anti-peroxidase(GOD-PAP) was used to check the levels of fasting blood-glucose.Cholesterol oxidase method was used to check the levels of total cholesterol(T-CH). Glycerol-phosphate oxidase was used to check the levels of triglyceride(TG). Enzymes colorimetric method was used to check high density level of lipoprotein cholesterol(HDL-C) and low density level of lipoprotein cholesterol (LDL-C), and the height, weight,waist circumference, and hip circumference were measured. Body mass index (BMI) and insulin resistance index (HOMA-IR) were calculated. Groups were compared using t test and correlation analysis using Pearson correlation test, and the relationship between serum resistin level and blood sugar, blood lipids and thyroid hormone were analyzed using multiple linear stepwise regression analysis method. Results Serum glucose[(5.2 ± 0.7)mmol/L],resistin concentrations[(132.1 ± 41.3)μg/L], FT3[(19.8 ± 8.7) pmol/L], FT4[(54.1± 29.6)pmol/L], fasting insulin levels[(7.9 ± 2.8)mU/L] and HOMA-IR(2.3 ± 1.0) were significantly higher compared with controls group [(4.7 ± 0.5)mmol/L, (65.1 ± 5.9)μg/L, (4.1 ± 0.6)pmol/L, (14.3 ± 2.2)pmol/L, (6.4 ± 2.7)mU/L, (1.5 ±1.2); t =4.64, 10.17, 11.42, 8.49, 4.48, 9.42, P＜ 0.01 or ＜ 0.05)]. T-CH[(3.7 ± 0.8) mol/L], LDL-C[(1.8 ±0.6)mol/L], TSH[(0.01 ± 0.01 )mU/L] were significantly lower compared with controls group[(4.6 ± 0.7)mol/L,(2.3 ± 0.7)mol/L, (1.80 ± 0.90)mU/L; t =5.30, 3.33, 14.48; all P＜ 0.01)]. Pearson correlation analysis showed that resistin and FT3, FT4 and HOMA-IR was positively correlated, respectively (r =0.719, 0.790, 0.396, P ＜ 0.01or＜ 0.05), resistin and T-CH and LDL was negatively correlated, respectively(r =- 0.364, - 0.519, P ＜ 0.05or ＜ 0.01). Multiple linear regression analysisshowed that resistin and FT3, FT4, and HOMA-IR was positively correlated, respeetively(r =0.756, P ＜ 0.01 ). Conclusion Resistin and FT3 and FT4 are related. Resistin might play important roles in insulin resistance and glucose and lipid metabolism disorders in patients with hyperthyriodism.

Significance of Galectin-3 and HBME-1 expression in differential diagnosis of thyroid nodules / 中国地方病学杂志

Objective To study the difference of galectin-3 and HBME-1 expression between benign and malignant thvreid nodules and to evaluate their clinical value.Methods Using immunohistochemical SP method,the paraffin-embedded tissues of 30 ewes papillary thyroid carcinoma(PTC)and 30 cases nodular goiter(NG) were examined to deteet the expression of Galectin-3 and HBME-1.Results The rates of positive staining of Galectin-3.HBME-1.Galectin-3 combined with HBME-1 in PTC were 93.3%(28/30),83.3%(25/30)and 100.0%(30/30),respectivdy,were higher than that of NG[20.0%(6/30),6.7%(2/30)and 26.7%(8/30)].There were significant differences between PTC and NG(X2＝32.85,35.65,34.74,all P＜0.01).The sensitivity,specificity,positive predietive value.negative predictive value and accuracy rate of using Galectin-3,HBME-1,Galectin-3 combined with HBME-1 in differentiation NG from FrIE were 93.3%,83.3%,100.0%;80.0%,93.3%,73.3%;82.3%,92.6%,78.9%:92.3%,84.8%,1 00%;86.7%,88.3%,86.7%,respectively.Conclusion Immunohistochemical stains of Galectin-3 and HBME-1 may be used as two lnarkel's for differentiating PTC and NG,it is especially sensitive when the two markers are jointly used.

Association of E23K polymorphism of inwardly rectifying K~+channel 6.2 gene with the phenotype of type 2 diabetes and glucose-lowering effect of gliclazide / 中华内分泌代谢杂志

The influences of E23K polymorphism of inwardly rectifying K~+channel 6.2 (Kir6.2) gene on the clinical phenotype of type 2 diabetes and glucose-lowering effect of gliclazide were investigated.The result showed that E23K polymorphism did not influence glucose-lowering effect of gliclazide,but serum creatinine level of patients with K/K genotype was higher than those of E/E and E/K genotypes (P