Objective To explore the clinical characteristics of patients with diffuse large B-cell lymphoma (DLBCL) accompanied with KMT2D gene mutation and the impact of its co-mutated genes on prognosis. Methods Clinical data of 155 newly diagnosed DLBCL patients were obtained. The second-generation sequencing method was used to detect 475 hotspot genes, including KMT2D mutation. Patients were divided into the KMT2D mutation group and KMT2D wild-type group based on the presence or absence of KMT2D gene mutation. Clinical characteristics, differences in co-mutated genes, and survival differences between the two groups were compared. Results The frequency of KMT2D mutation was 31%, which is predominantly observed in elderly patients (P=0.07) and less in the double-expressor phenotype (P=0.07). Compared with the KMT2D wild-type group, KMT2D gene mutation was associated with higher co-mutation rates of CDKN2A (OR=2.82, P=0.01) and BCL2 (OR=3.84, P=0.016), while being mutually exclusive with MYC gene mutation (OR=0.11, P=0.013). In univariate survival analysis, no statistically significant difference in overall survival (OS) was found between the KMT2D mutation group and the wild-type group (P=0.54). Further analysis of the prognostic significance of KMT2D with other gene mutations indicated that patients with KMT2D^mutBTG2^mut had poorer OS than those with KMT2D^wt BTG2^mut (P=0.07) and KMT2D^wt BTG2^wt (P=0.05). On the contrary, patients with KMT2D^mut CD79B^mut had better OS than those with KMT2D^mut CD79B^wt (P=0.09), with no prognostic impact observed for other co-mutated genes. Multivariate Cox regression analysis revealed that Ann Arbor stages Ⅲ and Ⅳ (HR=2.751, 95%CI: 1.169-6.472, P=0.02), elevated LDH levels (HR=2.461, 95%CI: 1.396-4.337, P=0.002), Ki-67 index>80% (HR=1.875, 95%CI: 1.066-3.299, P=0.029), and KMT2D^mutBTG2^mut(HR=4.566, 95%CI: 1.348-15.471, P=0.015) were independent risk factors for OS in patients with DLBCL (P<0.05). Conclusion DLBCL patients with KMT2D mutation often have multiple gene mutations, among which patients with a co-mutated BTG2 gene have poor prognosis.

To investigate the clinical features and prognostic significance in myelodysplastic syndrome (MDS) patients with DTA (DNMT3A,TET2,ASXL1) mutations. Clinical characteristics of 140 patients diagnosed as de novo MDS at People′s Hospital of Xinjiang Uygur Autonomous Region from September 2015 to December 2019 were retrospectively analyzed. Next-generation sequencing was used to detect 34 related genes in MDS patients. DTA mutations and the correlation with progression-free survival (PFS) and overall survival (OS) in MDS patients were evaluated. Among 140 MDS patients, DTA mutations was detected in 62 (44.3%) patients. And the positive rate of DTA mutations in IPSS-R lower-risk group was 65.4%, significantly higher than that of higher-risk group (31.8%)( P=0.000). Compared with the non-mutated group, patients with DTA mutations had a lower rate of conversion to leukemia (9.7% vs . 29.5%, P=0.004).Survival analysis showed that PFS in patients with DTA mutations was comparable as that in MDS patients without DTA mutations ( P=0.787), but the median OS was significantly shorter (16 months vs . 20 months, P=0.022).According to IPSS-R classification, the median OS in patients with and without DTA mutation was only statistically significant in the higher-risk group (15 months vs. 18 months, P=0.034).Among 62 patients with DTA mutations, 60 (96.8%) had additional gene mutations. DTA mutations were not independent prognostic factors when mutation frequency is greater than 10% were considered in Cox regression model ( P>0.05). DTA mutations often developed in the early stage of MDS, therefore they were more common in IPSS-R lower-risk subgroup which was correlated to the low rate of conversion to leukemia. In conclusion, DTA mutations are not associated with disease progression, but predict unfavorable survival when other add-on genes are mutated.

Objective:To investigate the expression levels of CD4+ CD25+ FOXP3+ Treg cells and interleukin(IL)-10 in serum of patients with myelodysplastic syndrome (MDS) - refractory anemia (RA) and refractory hematopenia with multilineage dysplasia (RCMD), and to evaluate the effect of cyclosporine on CD4+ CD25+ FOXP3+ Treg cells in MDS patients.Methods:From January 2016 to January 2018, 25 MDS-RA and RCMD patients and 13 healthy controls were selected from people′s Hospital of Xinjiang Uygur Autonomous Region for retrospective analysis.The expression of CD4 + CD25 + Foxp3 + Treg and IL-10 in peripheral blood samples were detected by flow cytometry and enzyme-linked immunosorbent assay.The expression of CD4 + CD25 + Foxp3 + Treg and IL-10 in MDS-RA and RCMD patients before and 6 months after the treatment with CSA based immunosuppressive regimen was detected.Results:Of the 25 patients, 13 (52%) were effective and 12 (48%) were ineffective.The proportion of CD4 + CD25 + Foxp3 + Treg in CD4 + T cells of MDS group was significantly higher than that of healthy control group [(0.37 ± 0.10)% and (0.12 ± 0.06)% respectively, t= 2.02, P< 0.001]. The level of IL 10 in MDS group was significantly higher than that in healthy control group ((7.16±1.27) μg /L and (2.75 ± 1.06) μg /L, t= 2.03, P< 0.001). The ratio of CD4 + CD25 + Foxp3 + Treg cells in MDS group was lower than that in MDS Group ((0.15±0.06)% and (0.26±0.08%), t= 1.71, P< 0.001), and the level of IL 10 in MDS group was lower than that in MDS Group ((3.22±1.01) μg /L and (4.25±1.22) μg /L, t= 2.06, P= 0.030). The proportion of CD4 + CD25 + Foxp3 + Treg in peripheral blood of 25 MDS patients was positively correlated with the level of IL-10 expression ( r= 0.35, P= 0.02). Conclusion:The expression of CD4+ CD25+ FOXP3+ Treg cells and IL-10 increased in MDS patients increased, but decreased after cyclosporine treatment.

To investigate the expression of microRNA-34a (miR-34a) in patients with chronic lymphocytic leukemia (CLL) in Xinjiang Uygur and Han nationalities and its prognostic significance. Our data showed that miR-34a expression in Uygur and Han CLL patients was significantly higher than that in their respective healthy controls, while miR-34a levels were similar between Uygur and Han patients. By comparing with known prognostic factors, receiver operating characteristic (ROC) curves showed that miR-34a was a good predictive factor for the prognosis of CLL (demarcation value was 3.567 6). Survival analysis was further performed according to miR-34a expression level, that low expression of miR-34a translated into poor prognosis.

Objective To explore the clinical and laboratory characteristics of chronic B lymphocyte proliferation disease (B-CLPD) without typical lymphoid proliferation. Methods The clinical records of patients with B-CLPD only characterized by pancytopenia form January 2007 to March 2016 in hematology department of Xinjiang Uygur Autonomous Region People ' s Hospital were collected, and the cell morphology, bone marrow pathology, cytogenetics and molecular characteristics were retrospectively analyzed. Results The median age of 11 patients was 68 years old. The lymphocyte ratio of peripheral blood smears in all patients increased in different level (0.36-0.68), but absolute lymphocyte count was normal or decreased (0.59×109-1.99×109). Lymphocyte-like plasma cell or small numbers of plasma cell can be seen in the bone marrow smears of 4 cases and lymphocytes with irregular burr-like protrusions were observed in 2 cases while there were no characteristic morphological changes in remained 5 cases. Immunophenotypical analysis showed that all patients expressed CD19, CD20, CD22, SmIg, not expressed CD5, CD10 which with scores of 0-2 according to chronic lymphocytic leukemia (CLL) points system;CD103, CD11C, CD25 and FMC7 were highly expressed in 2 cases while there were no characteristic expression in remaining cases. There were no abnormal karyotypes observed from the conventional cytogenetic and fluorescence in situ hybridization (FISH) analysis (both of IgH/CCND1, bcl-2/IgH were negative) in all patients. 8 patients were found IgH gene rearrangement, MYD88L256P and BRAF V600E was positive in 5 cases and 2 cases respectively. 5 cases were diagnosed as Waldenstrom macroglobulinemia, 3 cases were B-CLPD, 2 cases were hairy cell leukemia, 1 case was nodal marginal zone B-cell lymphoma after comprehensive analysis of their clinical and laboratory data. Conclusion Even if there are no increased peripheral blood lymphocytes in pancytopenia patients, it is necessary to perform bone marrow smears, immunophenotyping, IgH gene rearrangement, cytogenetics and other molecular laboratory tests to exclude B-CLPD, and reduce misdiagnosis.

Objective To investigate the prognostic significance of serum C-reactive protein (CRP) in diffuse large B-cell lymphoma (DLBCL).Methods 106 cases of newly diagnosed DLBCL patients with average age of 55.23 years old were analyzed retrospectively.Determine the level of serum CRP and other factors,including tumour stage,revised Internation Prognostic Index (R-IPI) before the treatment.After median follow-up of 62 months,the relationships between serum CRP and overall survival (OS) and event-free survival (EFS) were observed.Results 106 DLBCL patients were divided into 4 Ann Arbor stages.Median serum CRP levels significantly increased in higher Ann Arbor stages [stage Ⅰ (9.49±2.73) mg/L,stage Ⅱ (14.96± 2.60) mg/L,stage Ⅲ (22.03±3.39) mg/L,stage Ⅳ (28.96±4.56) mg/L] (all P ＜ 0.05).The serum CRP level was positively correlated with tumor stage (r =0.833,P =0.000),R-IPI (r =0.344,P =0.000),and no correlated with age (r =-0.118,P ＞ 0.05),sex (r =0.085,P ＞ 0.05).The patients with serum CRP levels ＞19.0 mg/L had shorter 5-year ESF and OS than patients with serum CRP levels ≤ 19.0 mg/L (P ＜ 0.05).Univariate and multivariate Cox regression models showed that old age,high R-IPI score and high CRP level were independent adverse prognostic factors.Conclusion In DLBCL patients,high CRP level not only reflects tumor burded,but also delivers prognostic information.

OBJECTIVETo evaluate the significance of T-cell antigen receptor (TCR) gene rearrangement among patients with acquired pure red cell aplastic anemia (A-PRCA).

METHODSFor 16 patients with A-PRCA, an immunosuppressive regimen based on cyclosporin A (CsA) was applied. Rearrangement of the TCR gene was detected by PCR, and T lymphocyte subsets in peripheral blood specimens was detected with flow cytometry.

RESULTSFive patients had presented with TCR clonal rearrangement and were positive for both TCR γ and TCR δ. The blood of 13 patients have returned to normal with the treatment, which included 3 cases with bone marrow returning to normal. In 7 cases, the red cell hyperplasia of bone marrow is still down, but has increased with the treatment. Three patients were close to cure, 7 showed remission, 3 were improved, but 3 were ineffective. The rate of effective treatment in those with TCR rearrangement (2/5) was significantly lower than that those without (11/11, chi-square=8.123, P < 0.05). Compared with those without the TCR gene rearrangement, the Th cells and proportion of Th/Ts were significantly lower, while the Ts cell (CD3+CD8+) were significantly higher in those with the rearrangement (P < 0.05).

CONCLUSIONTCR gene rearrangement may play a role in the pathogenesis of A-PRCA. CsA is effective for the treatment of A-PRCA, but patients presenting clonal TCR gene rearrangement may response poorly to the treatment.

Adult ; Aged ; Female ; Gene Rearrangement, T-Lymphocyte ; Humans ; Male ; Middle Aged ; Red-Cell Aplasia, Pure ; etiology ; genetics ; immunology

Objective To detect and analyse acquired pure red cell aplasia (PRCA) T lymphocyte subsets distribution and to assess its condition and cyclosporine immune function of T lymphocytes subsets.Methods Flow cytometry was applicated to detect peripheral blood T lymphocyte subsets of acquired PRCA patients before and after 3 months of treatment with cyclosporine-based immunosuppressive regimen and normal controls.Results Among 22 patients,17 cases of blood returning normal (three cases of bone marrow returning normal),the total effective rate was 95.5 ％ (3 cases of cure,14 cases of remission,4 cases of improvement,1 case of ineffectiveness).Th (CD3+ CD4+) cells and Th/Ts ratios in acquired PRCA patient group were lower than those in the normal control group,while Ts (CD3+ CD8+) cells was higher than that in the normal control group,the differences were statistically significant (P ＜ 0.05).After 3 months of treatment,Th cells and Th/Ts ratio were higher than those before,and Ts cells were decreased compared with the previous (P ＜ 0.05).Conclusion Disorders of T lymphocyte subsets in acquired PRCA patients lead to immune dysfunction,however,cyclosporine can improve T lymphocyte subsets in patients with imbalance,which is an effective way to treat the disease with significant curative effect and mild adverse reactions.

Objective To investigate the characteristics of chromosome kayotypes and the relationship between the prognosis and chromosome karyotypes in subtypes of myelodysplastic syndromes (MDS).Methods The study retrospectively analyzed the characteristics of chromosome karyotype of initially diagnosed 151 MDS patients and investigated the rate and time of leukemia transformation and mortality,IPSS score,and compare the ethnic differences of Han and Uyghurs.Results Abnormal karyotype detection rate was 55.0 ％ (83/151),including simple abnormalities was 53.0 ％ (44/83),complex abnormalities was 47.0％ (39/83).h showed that common abnormal karyotype include-5/5q-,-7/7q-,+8,-20/20q-,-X/-Y,i(17q),9p-/9q-,+21.IPSS score had differences among subtypes (x2 =117.802,P ＜ 0.01).The detection rates of abnormal chromosome had significantly differences between each group,the abnormal karyotype detection rate in high-risk group was significantly higher than those in low risk group and moderate group(P ＜ 0.05).Followup 31 months (5-68 months) and found that the rates of leukemia transformation and mortality were 25.2 ％ (38/151) and 43.7 ％ (66/151),the rates of leukemia transformation and mortality in abnormal karyotype group were significantly higher than those in normal karyotype group (P ＜ 0.05).The median survival time in abnormal karyotype was shorter than that in normal one.The distribution of Han and Uyghur patients with MDS subtypes,the characteristics of abnormal karyotype,the rates of leukemia transformation and the rates of mortality had no statistical difference (all P ＞ 0.05).Conclusion Abnormal chromosome karyotype is important index for disease progression and prognosis of MDS patients.

Objective To investigate the characteristics of the abnormal karyotype and normal karyotype with myelodysplastic syndrome(MDS).Methods A retrospective analysis of 131 MDS patients was conducted.The cell morphology between abnormal karyotype and normal karyotype was compared.Results Of 131 MDS patients,71 cases (56.5％)had clonal chromosomal abnormalities.Pelger nuclear myeloid and lymphoid small megakaryocytes in abnormal karyotype group was significantly higher than the normal karyotype group (P ＜ 0.05).Megaloblastic erythroid-like change,double-nucleated red blood cells,multinucleated red blood cells,the petals nuclear,nuclear fragmentation;the myeloid uneven particle distribution,nuclear pulp imbalance,megaloblastic degeneration,vacuoles,AUER,dual-core; single-round,multi-roundnuclear megakaryocytes,the two groups showed no significant differences (P ＞0.05).Conclusion Pelger nuclear myeloid,lymphoid small megakaryocytes had significantly higher incidence of abnormal karyotype MDS compared with normal karyotype cell dysplasia,there was some correlation between abnormal karyotype and cell morphology.