A neural pathway for social modulation of spontaneous locomotor activity (SoMo-SLA) in Drosophila.

Social enrichment or social isolation affects a range of innate behaviors, such as sex, aggression, and sleep, but whether there is a shared mechanism is not clear. Here, we report a neural mechanism underlying social modulation of spontaneous locomotor activity (SoMo-SLA), an internal-driven behavior indicative of internal states. We find that social enrichment specifically reduces spontaneous locomotor activity in male flies. We identify neuropeptides Diuretic hormone 44 (DH44) and Tachykinin (TK) to be up- and down-regulated by social enrichment and necessary for SoMo-SLA. We further demonstrate a sexually dimorphic neural circuit, in which the male-specific P1 neurons encoding internal states form positive feedback with interneurons coexpressing doublesex (dsx) and Tk to promote locomotion, while P1 neurons also form negative feedback with interneurons coexpressing dsx and DH44 to inhibit locomotion. These two opposing neuromodulatory recurrent circuits represent a potentially common mechanism that underlies the social regulation of multiple innate behaviors.

A pilot study of orelabrutinib treatment in three cases of refractory/relapsed autoimmune haemolytic anaemia/Evans syndrome.

Currently, there is no effective treatment for refractory/relapsed (R/R) autoimmune haemolytic anaemia (AIHA), associated with poor quality of life. Bruton tyrosine kinase inhibitors have begun to be used in some autoimmune diseases. We initiated the clinical trial of orelabrutinib treatment on R/R AIHA/Evans Syndrome, which is in progress. The preliminary results showed that nine of the 12 enrolled patients responded to orelabrutinib treatment. Here, we reported three cases who have completed the treatment and were followed up for 6 months, achieving complete or partial remission. Orelabrutinib is expected to become a new second-line treatment for R/R AIHA/Evans syndrome.

Thymocyte selection-associated high mobility box protein regulates T lymphocytes exhaustion in patients with myelodysplastic syndromes by inhibiting PI3K/AKT/mTOR pathway.

Myelodysplastic syndromes (MDS) patients often experience CD8+ T lymphocytes exhaustion, which plays a crucial role in the development of MDS. However, the specific role of thymocyte selection-associated high mobility box protein (TOX) in the CD8+ T lymphocytes exhaustion in MDS patients remains unclear. In this study, we investigated the role of TOX in CD8+ T lymphocytes exhaustion in patients with MDS. The expression of TOX, inhibitory receptors (IRs), and functional molecules in peripheral blood T lymphocytes of MDS patients and normal controls were detected using flow cytometry. Lentiviral transduction was used to create stable TOX-knockdown CD8+ T lymphocytes, and small interfering RNA (si-RNA) was used to knock down TOX in Jurkat cells. The expression of TOX was found to be significantly higher in CD8+ T lymphocytes of MDS patients compared to normal controls. This was associated with upregulated IRs and reduced expression of functional molecules such as Granzyme and Perforin. Myelodysplastic syndromes patients with higher TOX expression had poor clinical indicators and shorter survival. Knockdown of TOX using sh-RNA partially reverses the exhausted phenotype and enhances the lethality of CD8+ T lymphocytes. Moreover, the knockdown of TOX using si-RNA in Jurkat cells improved cell proliferation activity, down-regulated IRs and activated PI3K/AKT/mTOR signaling pathway. TOX promotes the exhaustion of CD8+ T lymphocytes by inhibiting PI3K/AKT/mTOR pathway, and targeted inhibition of TOX could partially restore the effector functions and activity of CD8+ T lymphocytes.

Increased ferroptosis of erythrocytes is associated with myelodysplastic syndromes.

Anemia is the most common symptom in patients with myelodysplastic syndromes (MDS). Programmed cell death of erythrocytes is one of the contributing factors to anemia. Ferroptosis is a newly identified form of iron-dependent cell death. The aim of this study is to investigate whether anemia in MDS patients is associated with ferroptosis of nucleated erythrocytes(NEs).We detected lipid peroxidation levels, Fe2+ contents, cell death rates, glutathione (GSH) and malondialdehyde (MDA) levels in bone marrow CD235a+ NEs of MDS patients. Expression levels of ferroptosis-related molecules (ACSL4, GPX4, and SLC7A11) were evaluated through qRT-PCR and Western Blotting. Correlation between these markers and clinical parameters were analyzed. To further substantiate that the mode of cell death with CD235a+ NEs of MDS patients was attributed to the ferroptosis pathway, we applied Fer-1 to inhibit ferroptosis. Cell viability was assessed using CCK8, and changes in ferroptosis-related indicators were simultaneously evaluated. We discover that the ferroptosis level of bone marrow NEs in MDS patients was increased, which is related to anemia and iron overload. Ferroptosis might be one of the causes of anemia in MDS patients.

A nomogram model for predicting the efficacy of cyclosporine in patients with pure red cell aplasia.

Pure red cell aplasia (PRCA) is a rare bone marrow disorder characterized by a severe reduction or absence of erythroid precursor cells, without affecting granulocytes and megakaryocytes. Immunosuppressive therapies, particularly cyclosporine, have demonstrated efficacy as a primary treatment. This study aims to develop a predictive model for assessing the efficacy of cyclosporine in acquired PRCA (aPRCA). This retrospective study encompasses newly treated aPRCA patients at the General Hospital of Tianjin Medical University. Diagnosis criteria include severe anemia, and absolute reticulocyte count below 10 × 109/L, with normal white blood cell and platelet counts, and a severe reduction in bone marrow erythroblasts. Cyclosporine therapy was administered, with dose adjustments based on blood concentration. Response to cyclosporine was evaluated according to established criteria. Statistical analysis involved logistic multi-factor regression, generating a predictive model. The study included 112 aPRCA patients with a median age of 63.5 years. Patients presented with severe anemia (median Hb, 56 g/L) and reduced reticulocyte levels. Eighty-six patients had no bone marrow nucleated erythroblasts. Primary PRCA accounted for 62 cases (55.4%), and secondary PRCA accounted for 50 cases (44.6%). Univariate analysis revealed that ferritin, platelet to lymphocyte ratio (PLR), and CD4/CD8 ratio influenced treatment response. Multivariate analysis further supported the predictive value of these factors. A prediction model was constructed using ferritin, PLR, and CD4/CD8 ratio, demonstrating high sensitivity and specificity. The ferritin, PLR, and CD4/CD8-based nomogram showed good predictive ability for aPRCA response to cyclosporine. This model has potential clinical value for individualized diagnosis and treatment of aPRCA patients.

High TOX expression on CD8+ T cells in pure red cell aplasia.

Thymocyte selection-associated high-mobility group box protein (TOX) is an important molecule regulating the development and exhaustion of T lymphocytes. Our aim is to investigate the role of TOX in the immune pathogenesis of pure red cell aplasia (PRCA). TOX expression of CD8+ lymphocytes from the peripheral blood of patients with PRCA was detected by flow cytometry. Additionally, the expression of immune checkpoint molecules PD1 and LAG3 and cytotoxic molecules perforin and granzyme B of CD8+ lymphocytes was measured. The quantity of CD4+CD25+CD127low T cells was analyzed. TOX expression on CD8+ T lymphocytes in PRCA patients was significantly increased (40.73 [Formula: see text] 16.03 vs. 28.38 [Formula: see text] 12.20). The expression levels of PD1 and LAG3 on CD8+ T lymphocytes in PCRA patients were significantly higher than those in the control group (34.18 [Formula: see text] 13.26 vs. 21.76 [Formula: see text] 9.22 and 14.17 [Formula: see text] 13.74 vs. 7.24 [Formula: see text] 5.44, respectively). The levels of perforin and granzyme in CD8+ T lymphocytes of PRCA patients were 48.60 [Formula: see text] 19.02 and 46.66 [Formula: see text] 25.49, respectively, which were significantly higher than those of the control group (31.46 [Formula: see text] 7.82 and 16.17 [Formula: see text] 4.84, respectively). The number of CD4+CD25+CD127low Treg cells in PRCA patients was significantly decreased (4.30 [Formula: see text] 1.27 vs. 1.75 [Formula: see text] 1.22). In PRCA patients, CD8+ T cells were activated and exhibited overexpression of TOX, PD1, LAG3, perforin, and granzyme B, while regulatory T cells decreased. These findings suggest that T cell abnormality plays a critical role in the pathogenesis of PRCA.

The relationship between NICU stress and neurodevelopmental outcomes of preterm infants: A multi-center prospective cohort study in China.

PURPOSE: To examine the relationship between NICU stress exposure and the neurodevelopmental outcomes of preterm infants. DESIGN AND METHODS: A multicenter, prospective cohort study was conducted between May 2021 and June 2022. Preterm infant participants (28-34 weeks gestational age) were recruited at birth from three NICUs of three tertiary hospitals by convenience sampling. The NICU stress includes acute NICU stress and chronic NICU stress which were measured over the total NICU hospitalization for each infant using the Neonatal Infant Stressor Scale (NISS). Neurodevelopmental outcomes of preterm infants were assessed at 3 months corrected age (CA) using the Ages and Stages Questionnaire, Third Edition (ASQ-3). RESULTS: Of one hundred and thirty preterm infant participants, 108 preterm infants were included into analysis. Results showed that acute NICU stress exposure significantly predicted the neurodevelopmental abnormalities in communication function (RR: 1.001, 95%CI: 1.000-1.001, p = .011), while chronic NICU stress exposure was significantly associated with the problem-solving function (RR: 1.003, 95%CI: 1.001-1.005, p = .002) at 3 months CA. No significant associations were found between NICU stress exposure and other dimensions of neurodevelopmental outcomes, including gross motor, fine motor, and personal-social functions. CONCLUSION: NICU stress exposure demonstrated a significant predicting relationship with abnormalities in communication and problem-solving functions of preterm infants at 3 months CA. PRACTICE IMPLICATIONS: During the NICU hospitalization, neonatal health caregivers should systematically monitor the NICU stress exposure to prevent neurodevelopmental problems in preterm infants.

MicroRNA-21-5p Regulates CD3+T Lymphocytes Through VCL and LTF in Patients with Immune Thrombocytopenia.

BACKGROUND: Immune thrombocytopenia (ITP) is an autoimmune hemorrhagic disease with distinct clinical manifestations such as extensive skin petechiae, mucosal bleeding, and even visceral hemorrhage. In this study, CD3+T lymphocytes from ITP patients were screened for differentially expressed genes. The expression of miR-21 and miR-155 in T lymphocytes of ITP patients were investigated. The downstream target genes of miR-21 and miR-155 were also searched for the correlation between differentially expressed genes of ITP. METHODS: Differential gene screening was performed using the GSE43177 data set in the GEO database, and the expression of miR-21 and miR-155 in T lymphocytes of ITP patients was verified by qPCR. The interaction network of core downstream target genes and ITP differentially expressed genes of miR-21 and miR-155 were constructed with the STRING database, and the associated factors were verified by qPCR. RESULTS: In ITP patients, the expression of CD8+T lymphocytes increased, the expression of CD4+T lymphocytes decreased, and the ratio of CD4+/CD8+T cells decreased. Fourteen genes were differentially expressed in CD3+T lymphocytes, all of which were upregulated, and the expression of S100A8 was increased in ITP patients. The expression of miR-21-5p and miR-155-5p increased in CD3+T lymphocytes of initial ITP patients. The core down-stream target gene VCL of miR-21 was associated with the differentially expressed genes such as LTF, LCN2, and DEFA4 in the interaction network. VCL expression was decreased and LTF expression was increased in ITP patients. CONCLUSIONS: S100A8 plays an important role in the regulation of CD3+T lymphocytes in ITP patients. MiR-21-5p regulates the differentially expressed gene LTF by inhibiting the core downstream target gene VCL and participates in the immune mechanism of T lymphocytes in ITP patients. MiR-155-5p is also involved in the immunoregulatory mechanism of T lymphocytes in ITP patients.

Multiplex ligation-dependent probe amplification identifies copy number changes in normal and undetectable karyotype MDS patients.

Chromosomal abnormalities play an important role in classification and prognostication of myelodysplastic syndrome (MDS) patients. However, more than 50% of low-risk MDS patients harbor a normal karyotype. Recently, multiplex ligation-dependent probe amplification (MLPA) has emerged as an effective and robust method for the detection of cytogenetic aberrations in MDS patients. To characterize the subset of MDS with normal karyotype or failed chromosome banding analysis, we analyzed 144 patient samples with normal karyotype or undetectable through regular chromosome banding analysis, which were subjected to parallel comparison via fluorescence in situ hybridization (FISH) and MLPA. MLPA identifies copy number changes in 16.7% of 144 MDS patients, and we observed a significant difference in overall survival (OS) (median OS: undefined vs 27 months, p=0.0071) in patients with normal karyotype proved by MLPA versus aberrant karyotype cohort as determined by MLPA. Interestingly, patients with undetectable karyotype via regular chromosome banding indicated inferior outcome. Collectively, MDS patients with normal or undetectable karyotype via chromosome banding analysis can be further clarified by MLPA, providing more prognostic information that benefit for individualized therapy.

Clinical study on empirical and diagnostic-driven (pre-emptive) therapy of voriconazole in severe aplastic anaemia patients with invasive fungal disease after intensive immunosuppressive therapy.

The aim of this study is to compare the curative effect of empirical with diagnostic-driven (pre-emptive) therapy of voriconazole in severe aplastic anaemia patients (SAAs) with invasive fungal disease (IFD) after intensive immunosuppressive therapy (IST). Patients undergoing voriconazole antifungal therapy were randomized to empirical therapy group and diagnostic-driven therapy group. Empirical therapy group accounted for 48.5% of all cases, and diagnostic-driven therapy group accounted for 51.5%. The morbidity of IFD (probable and proven cases) was slightly increased in diagnostic-driven therapy group compared with empirical therapy group (P > 0.05). The total effective rate was 62.1%. The effective rate in empirical therapy group was 78.1%, which was significantly increased compared with diagnostic-driven therapy group (47.1%) (P < 0.05). This value was especially significant in possible IFD cases (P < 0.05). The efficacy of possible IFD cases in empirical therapy group was the best (84%) followed by the probable and proven cases in empirical therapy group (57.1%). In diagnostic-driven therapy group, the efficacy of possible IFD cases was 50%, and the efficacy of probable and proven cases was only 37.5%. The difference was statistically significant (P < 0.05). Absolute neutrophil count (ANC) is the key anti-infection factor. The efficacy of patients whose ANC ˂ 0.1 × 109/L was 39.28%, which was significantly reduced compared with that of patients whose ANC ≥ 0.1 × 109/L (78.95%) (P < 0.05). This finding was especially obvious in diagnostic-driven therapy group. As empirical therapy is superior to diagnostic-driven therapy, we recommend that empirical therapy should be started for high-risk patients, and efforts should be made to definitively diagnose the disease.

Abnormal Macrophage Polarization in Patients with Myelodysplastic Syndrome.

BACKGROUND: This study is aimed at assessing the subsets of bone marrow macrophages in patients with myelodysplastic syndrome (MDS) and exploring the role of macrophages in the pathogenesis of MDS. METHODS: Thirty-eight newly diagnosed MDS patients were enrolled in the Department of Hematology of General Hospital of Tianjin Medical University from June 2015 to June 2016. Bone marrow monocytes and macrophage subsets (M1/M2) were detected in patients with MDS and normal controls by flow cytometry. M1 macrophages were cultured in vitro, and the expression of IL-1ß and TNF-α mRNA was measured using real-time polymerase chain reaction. RESULTS: Compared with the normal control group, the proportion of bone marrow monocytes was higher (2.11 ± 0.93% vs. 3.66 ± 3.38%), and the mean fluorescence intensity of surface molecule CD14 was lower in the higher-risk (HR) MDS group (639.05 ± 359.78 vs. 458.26 ± 306.72, p < 0.05). The ratio of M2 macrophages to monocytes was higher in patients with HR-MDS (1.82 ± 2.47% vs. 3.93 ± 3.81%, p < 0.05). The ratio of M1 to M2 macrophages was lower in the HR-MDS group (3.50 ± 3.22 vs. 1.80 ± 0.88, p < 0.05). The expression of IL-1ß and TNF-α mRNA in M1 macrophages was significantly lower in the MDS group (p < 0.05). CONCLUSIONS: Patients with MDS had abnormal macrophage polarization, which may be involved in the alteration of bone marrow microenvironments.

Immunological characteristics and effect of cyclosporin in patients with immune thrombocytopenia.

OBJECTIVE: Immune thrombocytopenia (ITP) is well-known as an antibody-mediated autoimmune disease, and it is easy to get response but often turns to relapse or refractory. Cyclosporin is a traditional immunosuppressant and had a good effect on ITP patients. In this paper, we retrospectively analyze the immunological characteristics and therapeutic effect of cyclosporin in 220 patients with ITP. METHODS: All newly diagnosed ITP patients in the Department of Hematology, Tianjin Medical University General Hospital from June 2018 to December 2020 were enrolled and divided into four groups according to the expression of autoantibodies and the occurrence of prodromal infection. The basic data and immune indexes of ITP patients in each group were collected. The clinical immunological characteristics of patients in each group and the therapeutic effect of cyclosporin in each group were analyzed. RESULTS: Multi-autoantibody ITP patients were more likely to have low serum albumin and high gamma globulin, and the ratio of albumin to globulin decreased. In addition, the level of IgA and IgG increased and the level of complement C3 and C4 decreased more frequently than those in other groups. The number of CD3+T lymphocytes, especially CD3+CD4+T lymphocytes, decreased in ANA+ITP patients. The number of CD16+CD56+NK cells, pDC/DC ratio, and pDC/mDC ratio were higher than those in other groups. The expression of IL-6 and the proportion of CD19+B lymphocytes increased in two groups of ITP patients with abnormal autoantibodies. The patients of pro-infected group were more likely to suffer from coagulation disorder. After treatment with cyclosporin, the response rate increased and the 3-month relapse rate decreased in all ITP patients, and the therapeutic effect of patients with high megakaryocyte number was significantly higher than that of patients with low megakaryocyte number. The impact factors that influence the effect of glucocorticoid and(or) IVIG were the number of CD3+CD8+T lymphocytes, CD4/CD8 cell ratio, and the number of CD19+B lymphocytes. The independent impact factor of cyclosporin therapeutic response rate was the number of CD3+T lymphocytes. CONCLUSIONS: ITP is a heterogeneous disease, recurrence may occur during or rapidly after treatment. Cyclosporine included treatment can improve the effective rate of ITP and reduce the relapse rate within 3 months. The number of CD3+T lymphocytes was the only impact factor that influence the therapeutic effect of cyclosporin in ITP patients.

Analysis of clinical characteristics of 92 patients with paroxysmal nocturnal hemoglobinuria: A single institution experience in China.

OBJECTIVES: We performed a retrospective analysis to investigate the clinical characteristics and therapeutic strategies of Chinese paroxysmal nocturnal hemoglobinuria (PNH) patients, and assessed the efficacy and safety of glucocorticoid in PNH patients. METHODS: The clinical data of 92 PNH cases in our hospital were analyzed, including clinical manifestation, laboratory examination, treatment efficacy, and survival. RESULTS: The main clinical manifestations of these patients included hemoglobinuria, anemia, fatigue, dyspnea, headache, abdominal pain, and erectile dysfunction. Glucocorticoid is still the first-line treatment for PNH patients to control hemolytic attack, and the short-term remission rate (12 months) is 79.01% (64/81). Meanwhile, the overall survival (OS) of 10 years after diagnosis was estimated as 70.77% (46/65). Moreover, Cox proportional risk model for multivariate analysis showed that the increase in LDH multiple, thrombosis complications, and complicated with bone marrow failure were the independent adverse prognostic factors affecting the survival of PNH patients. CONCLUSION: Paroxysmal nocturnal hemoglobinuria patients in mainland China have various clinical features, while lower incidences of thrombosis and renal damage. Thrombosis and bone marrow failure are two complications with worse prognosis.

A Study of Immune Functionality of Newly Diagnosed Severe Aplastic Anemia Patients with Virus Infection.

BACKGROUND: Previous research showed that virus infection is correlated with the occurrence, development, and prognosis of AA. This study was designed to explore the influence of virus infection on the immune functionality and immunosuppressive therapy (IST) efficiency of newly diagnosed SAA patients. METHODS: Fifty-six newly diagnosed SAA patients combined with virus infection treated in the Hematology Department of Tianjin Medical University General Hospital from October 2004 to July 2014 were studied. Various immune parameters were tested and compared for SAA patients with and without virus infection. RESULTS: When compared with SAA patients without corresponding virus infection, SAA patients with CMV-IgM, PVB19-IgM, and EBV infection had increased CD8+ T cell percentage, decreased CD4+/CD8+ T cell ratios, and increased CD8+HLA-DR+/CD8+ percentage. The absolute value of CD8+ T cell of CMV-IgM group had increased as well. The CMV-IgM and PVB19-IgM groups showed decreased CD4+ T cell percentage, and decreased CD4+HLA-DR+/CD8+HLA-DR+ ratio. The PVB19-IgM group exhibited decreased CD4+HLA-DR+/CD4+ percentage, increased Th1 percentage and increased pDC percentage. Patients with EB virus infection showed lower NK cell percentage. Three years after IST, the treatment is significantly less effective for the SAA patients combined with virus infection than those without. CONCLUSIONS: CMV, PVB19, and EBV infection worsen the immune functionality abnormality of newly diagnosed SAA patients and reduce the IST efficiency.

[GDF11 level in patients with myelodysplastic syndrome and its clinical significance].

OBJECTIVE: To detect the expression level of growth differentiation factor 11 (GDF11) in patients with myelodysplastic syndrome (MDS), and to evaluate the relationship between GDF11 level and erythropoiesis functions. METHODS: A total of 44 MDS patients (18 low-risk group patients and 26 high-risk group patients) in Department of Hematology in Tianjin Medical University General Hospital and 10 normal controls were selected from September 2014 to June 2015. The concentration of GDF11 in peripheral blood was detected using enzyme-linked immunosorbent assay (ELISA). GDF11 mRNA expression in bone marrow mononuclear cells (BMMNC) was detected using RT-PCR method. The percentage of erythroid cells (CD235a) in bone marrow was detected by flow cytometry. The correlation between these indexes and erythropoiesis functions (including red blood cell count (RBC), hemoglobin level (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), reticulocyte percentage (Ret%), and proportion of nucleated eryhrocyte in bone marrow) were evaluated. RESULTS: (1) The GDF11 level in peripheral blood was significantly higher in high-risk group ((128.67±47.62) µg/L) than in low-risk group ((65.96±36.55) µg/L, P<0.01) and in normal controls ((29.76±10.10) µg/L, P<0.01), also significantly higher in low-risk group than in normal controls (P<0.05). The concentration of GDF11 in severe/moderate anemic MDS patients ((80.97±9.94) µg/L) was higher than that in normal controls/ mild anemic MDS patients((66.82±19.52) µg/L), but with no statistically significant (P>0.05). (2) The percentages of CD235a(+) cells in high-risk and low-risk groups were 38.49%±5.42% and 42.64%±7.36%, respectively, showing no statistically significant difference (P>0.05). (3) In high-risk group, the GDF11 level in peripheral blood was negatively correlated with Hb, RBC and Hct in peripheral blood (r=-0.437, -0.430, -0.306, all P<0.05), and positively correlated with nucleated eryhrocyte, Ret%, MCV and CD235a(+) cells in bone marrow (r=0.465, 0.392, 0.505, 0.387, all P<0.05), but not correlated with MCH and MCHC (both P>0.05). In low-risk group, the GDF11 level in peripheral blood was positively correlated with CD235a(+) cells in bone marrow (r=0.429, P<0.05), and not correlated with Hb, RBC, Ret%, MCHC, MCV, MCH, Hct and nucleated eryhrocyte (all P>0.05). (4) The mRNA expression of GDF11 in MDS patients (39.82±14.55) was higher than that in the controls (1.84±0.64, P<0.01). CONCLUSIONS: GDF11 level in peripheral blood is higher in patients with MDS and increases with the disease risk. The more severe the anemia, the higher the GDF11 level. It may be closely correlated with erythropoiesis indicators in MDS.

[CD22 signal abnormalities in the pathogenesis of immune related pancytopenia].

OBJECTIVE: To investigate the expression of CD22 and its downstream signal molecule spleen tyrosine kinase (SYK) and their phosphorylation of B lymphocytes in patients with immune related pancytopenia(IRP), and to explore the role of CD22 in pathogenesis of IRP. METHODS: The expression of CD22, SYK and their phosphorylation, along with the expression of IgG and IgM, which obtained from B lymphocytes in peripheral blood of 46 patients with IRP(22 new diagnosed and 24 remitted patients returned to normal after treatment), 22 healthy controls and 12 chronic lymphocytic leukemia(CLL) patients from February to December 2014 were analyzed by flow cytometry. And the mRNA expression of CD22 in peripheral blood mononuclear cell was determined by real-time quantitative PCR. RESULTS: The ratios of CD22+ cells and phosphorylated CD22(pCD22)+ cells of B lymphocytes in new diagnosed group (60. 03% ± 20. 94% 71. 32% ± 11. 16%) were significantly higher than those in remission group (46. 92% ± 20. 04%, 55. 82% ± 14. 42%), normal control group (46. 86% ± 17. 78%, 53. 28% ± 14. 76%) and CLL group (39. 74% ± 18. 96%, 59. 07% ± 17.09%) (all P <0.05). The ratios of phosphorylated SYK( pSYK) + cells in the four groups had the same trend (all P <0. 05). The ratio of pCD22+ cells/pSYK+ cells in new diagnosed group was significantly lower than that in normal control group and CLL group (27. 39 (5. 06 - 102. 70) vs 55. 95 (15. 25 - 298. 53), 56. 92(5. 60 - 228. 96), both P <0. 05), and pCD22+ cells positively correlated to pSYK+ cells ( r = 0. 341, P < 0. 05). The expression of IgG in new diagnosed group and remission group was significantly higher than that in normal control group, and the expression of IgM in new diagnosed group was significantly higher than that in normal control group and CLL group (all P <0. 05). The expression levels of CD22 mRNA in new diagnosed group was significantly higher than that in remission group, normal control group and CLL group (all P <0. 05). CONCLUSIONS: The BCR signal pathway of B lymphocyte in IRP patients is enhanced, and the quantity and function of CD22 are increased, while which are still insufficient to inhibit B cell proliferation, and these may have some relationships with the pathogenesis of IRP. [Key words] Pancytopenia; Antigens, CD22; Immune related pancytopenia; Spleen tyrosine kinase; Phosphorylation

[Clinical analysis on malignant clonal hematopoiesis in severe aplastic anemia patients with immunosuppressive therapy].

OBJECTIVE: To investigate the clinical characteristics and risk factors of monosomy 7 malignant clonal evolution in patients with severe aplastic anemia (SAA) treated with combined immunosuppressive therapy (IST). METHODS: The clinical data of SAA patients treated with IST who had monosomy 7 malignant clonal evolution from October 2004 to January 2012 were analyzed respectively. RESULTS: Six patients (4.2%) had monosomy 7 clonal evolutions. The median time to monosomy 7 was 36 (12-75) months after IST. All 6 patients were diagnosed myelodysplastic syndromes (MDS). Among them, 3 patients transformed to acute myeloid leukemia following MDS. The time was 24, 45 and 51 months after IST. The median following time was 42 (17-84) months. Four patients died during the following time. The median time from MDS to death was 9 (5-17) months. Among them, three patients died with infection, one died with cerebral hemorrhage. Six patients had the clinical characteristics that they had no response to IST after 6 months, high monocyte percentage in one month after IST combined with recombinant human granulocyte colony stimulating factor (rHu-GCSF) and agranulocytosis in 3 months after IST. CONCLUSION: Poor myeloid response to IST suggests malignant clonal hematopoiesis and poor prognosis in SAA patients.

[Clinical analysis of 70 cases of polycythemia vera].

OBJECTIVE: To analyze the clinical characteristics and laboratory data, treatment and prognosis of polycythemia vera (PV) and provide evidence for screening of high-risk population and looking for measures to reduce complecations. METHODS: A retrospective study was performed among 70 patients with PV from May 2005 to May 2014, 43 males and 27 females, aged (56.6±13.1) to collect the data about characteristics, laboratory data, myelogram chromosome, karyotypes, BCR/ABL and JAK2 V617F genes, as well as lactate dehydrogenase (LDH) and so on. RESULTS: At diagnosis, there were 42 cases (60.00%) had symptoms, 25 cases (35.71%) had thrombosis and embolism. Hemorrhage occurred in 3 cases (4.29%), splenomegaly in 48 cases (68.57%), hepatomegaly in 7 cases (10.00%). The average hemoglobin at diagnosis was 195.17 g/L, the white blood cells count was 10.12×10(9)/L, the platelet count was 295×10(9)//L. The chromosome karyotypes were all normal. The positive percentage of JAK2 V617F mutation was 87.69% (57/65). The disease outcomes were myelofibrosis for 3 paitents, death from ineffective treatment after transforming to myelofibrosis and then biphenotype acute leukemia for 1 patient, and death from cardiorespiratory failure for 2 patients. The level of erythropoietin in JAK2 V617F mutated group were significantly lower than those in wild-type JAK2 V617F group (P<0.05). The level of hemoglobin and platelet counts in JAK2 V617F mutated group were significantly higher than those in wild-type JAK2 V617F group (both P<0.05). CONCLUSION: PV is one of meyloproliferation neoplasm, characterized by abnormally increasing blood cells, thrombosis and transforming to other myeloproliferative neoplasms.

[Quantity and function of regulatory B cells of the patients with immune thrombocytopenia].

OBJECTIVE: To explore the quantity and function of regulatory B cells (Bregs) in peripheral blood (PB) of the patients with immune thrombocytopenia (ITP). METHODS: A total of 67 ITP patients (35 patients were newly diagnosed, 32 patients were in remission) were collected from November 2013 to October 2014 in our hospital. And 30 normal controls were enrolled in this study. Their PB Bregs (CD19⁺CD24(hi)CD38(hi) cells) and related molecules interleukin-10 (IL-10) in Bregs were detected by flow cytometry (FCM). Correlation between Bregs and clinical parameters of ITP patients was analysed. RESULTS: The percentage of Bregs in CD19⁺ cells of newly diagnosed ITP patients (3.09% ± 2.57%) was significantly lower than that of remitted ITP patients (7.78% ± 6.59%, P = 0.002) and controls (5.42% ± 3.31%, P = 0.003). The percentage of IL-10⁺ cells in Bregs of newly diagnosed ITP patients (32.30% ± 12.71%) was significantly lower than that of remitted ITP patients (71.73% ± 18.08%, P = 0.000) and controls (51.01% ± 17.08%, P = 0.026). And the percentage of IL-10⁺ cells in Bregs of remitted ITP patients was significantly higher than that of controls (P = 0.026). The level of Bregs of newly diagnosed ITP patients had significantly positive correlation with myeloid dendritic cells (mDC) (r = 0.644, P = 0.003) and PB platelet count (r = 0.327, P = 0.006). CONCLUSION: Bregs involved in the pathogenesis of ITP might be a potential therapeutic target of this disease.

[Clinical significance of serum bone metabolic markers in diagnosis and monitoring of myeloma bone disease].

OBJECTIVE: To explore the significance of serum bone metabolic markers in the diagnosis and monitoring of multiple myeloma bone disease(MBD). METHODS: Thirty-six newly diagnosed multiple myeloma (MM) patients who were treated in Department of Hematology, Tianjin Medical University General Hospital from January 2013 to December 2014 were collected. Bone morbidity was graded into two stages according to the radiographic evaluation of the skeleton: stage A (n=12) included patients with no lytic lesions or with osteoporosis alone; stage B (n=24) included patients with osteolytic lesions and/or a pathological fracture. All the patients achieved partial or complete remission after treated with bortezomib + dexamethasone + zoledronic acid regimen. A total of 25 aged- and gender-matched healthy individuals were enrolled in this study as controls. The levels of serum tartrate-resistant acid phosphatase isoform 5b (TRACP-5b), carboxy-terminal cross-linking telopeptide of type I collagen (CTX), osteocalcin (OCN), and procollagen I amino-terminal propeptide (PINP) were investigated by ELISA and electrochemiluminescence immunoassay (ECLIA). The differences of these bone metabolic markers before and after treatment, and at different stages of bone disease were observed. RESULTS: The value of TRACP-5b in the newly diagnosed MM was significantly higher than that in the healthy controls and after treatment(median 4.16 vs 2.63 U/L, P=0.014; 4.16 vs 2.61 U/L, P=0.037). Serum level of CTX in the newly diagnosed MM patients was significantly decreased after treatment (median: 0.26 vs 1.05 µg/L, P=0.003). The ratio of CTX/OCN and CTX/PINP decreased after treatment, but there were no significant differences (both P>0.05). The pretreatment level of serum TRACP-5b in stage B patients was higher than that of the healthy controls (median: 4.20 vs 2.63 U/L, P=0.015). The levels of serum CTX in stage A and stage B patients were both higher than that of the healthy controls (median: 1.16 vs 0.48 µg/L, P=0.002; 0.88 vs 0.48 µg/L, P=0.040). The levels of serum OCN and PINP were higher in stage A patients compared with stage B patients, but there were no significant differences (both P>0.05). The ratio of CTX/OCN and CTX/PINP of stage A and stage B patients all increased compared with those of the healthy controls, but there were no significant differences (all P>0.05). CONCLUSIONS: Bone damage of MM patients is improved after effective treatment, but bone imbalance still exists, indicating that the treatment of MBD is a long process. Abnormal serum levels of TRACP-5b and CTX are found before positive X-ray findings in MBD, suggesting that these biochemical markers could be used as indices for early diagnosis of MBD.