OBJECTIVE: To investigate the expression of CSF3R mutation in acute myeloid leukemia (AML) and analyze its clinical characteristics and prognosis. METHODS: A retrospective study was conducted in 212 patients with AML who were newly diagnosed in the Second Hospital of Shanxi Medical University from January 1th 2018 to June 30th 2021, including 22 patients with CSF3R mutations as mutation group and 190 patients with CSF3R wild type ［66 cases of them were screened by propensity score matching (PSM), as control group］. The early efficacy and survival between the two groups were compared. RESULTS: The median age of patients in the mutation group was 50(17-73) years old, and the ratio of male to female was 1.2:1 The main types were AML with maturation (11 cases) and acute myelomonocytic leukemia (9 cases). Prognostic stratification was carried out according to the risk stratification system of the European leukemia network in 2017, with 16 cases (72.73%) in the middle and high-risk group. At the initial diagnosis, the median count of white blood cell (WBC) was 44.75(1.30-368.71)×10⁹/L, among which 15 cases (68.18%) were >10×10⁹/L, and the median count of platelet (PLT) was 24(4-55)×10⁹/L. CSF3R T618I (68.18%) was a common mutation site, which had concomitant gene mutations, in which CEBPA mutation was the most common (10 cases, 45.45%), but only existed in CSF3R T618I mutation. The CR/CRi rate was 68.18% and 71.21% in the mutant group and the control group (P >0.05), the median over all survival time was 15 months and 9 months (P >0.05), and the median disease-free survival time was 8 months and 4 months (P >0.05), respectively. CONCLUSION Most AML patients with CSF3R mutation are middle-aged patients, the main types are AML with maturation and acute myelomonocytic leukemia, and most of them have middle and high-risk prognosis. CSF3R mutation may not be an independent prognostic marker for newly diagnosed AML patients.
Middle Aged ; Humans ; Male ; Female ; Aged ; Leukemia, Myelomonocytic, Acute ; Retrospective Studies ; Leukemia, Myeloid, Acute/diagnosis* ; Prognosis ; Mutation ; Receptors, Colony-Stimulating Factor/genetics*

OBJECTIVE: To investigate the difference of therapeutic effects on children with thalassemia at different age after hematopoietic stem cell transplantation. METHODS: The clinical data of children with thalassemia treated in our hospital were retrospectively analyzed. The children were divided into 2-5 years old group and 6-12 years old group. The success rate of implantation, transplant-related mortality, GVHD incidence, and other transplant-related complications, as well as thalassemia-free survival (TFS) were compared between the two groups. RESULTS: The incidence of GVHD, hemorrhagic cystitis and severe oral mucositis after transplantation in the 2-5 years old group were significantly lower than those in the 6-12 years old group, while there was no statistically significant difference in the TFS between the two groups. CONCLUSION Children in the low age (2-5 years old) group show fewer complications and higher quality of life after transplantation, therefore, stem cell transplantation at 2-5 years old is more conducive to rehabilitation of the children with thalassemia.
Child ; Child, Preschool ; Graft vs Host Disease/complications* ; Hematopoietic Stem Cell Transplantation ; Humans ; Quality of Life ; Retrospective Studies ; Thalassemia/therapy* ; beta-Thalassemia/therapy*

OBJECTIVE: To observe the safety of donor NK cell infusions in the settings of hematopoietic stem cell transplantation and after consolidation chemotherapy in elderly patients with acute myeloid leukemia (AML). METHODS: Forty patients with AML were included, in which 21 patients aged over 60 years were at the stage of complete remission (CR) and 19 patients that received allogeneic hematopoietic stem cell transplantation (allo-HSCT). Mononucleated cells were isolated from peripheral blood from the donors (for allo-HSCT) or healthy immediate family members (elderly AML). The cells were seeded into the flasks pre-coated with NK cell specific activators, and expanded in media containing recombinant human IL-15 and IL-2 for 14 days. The cells were transfused intravenously after the identification of quality control. Trypan blue exclusion test was used for the determination of cell viability and counting. Flow cytometry analysis was performed to assess the surface antigenic profile. Seventy-eight infusions of the cell products were received by the elderly patients with AML after consolidation chemotherapy, 11 infusions were received by the patients during allo-HSCT and 32 infusions 3 moths after transplantation. The safety of cell therapy, body temperature, blood pressure and other indexes were observe during and 48 hours after cell transfusion. Meanwhile, the occurrence and severity of acute graft-versus-host disease (GVHD) were documented. RESULTS: Flow cytometry analysis showed that the proportion of NK cells (CD3^-CD56⁺) in the mononucleated cells before culture was (14.10±4.22)% (n=121), and the proportion increased dramatically up to (87.29±8.75)% (n=121) after culture for 14 days, the number of NK cells increased to 753.47±140.13 times (n=121). The doses of the infused NK cells was (7.58±2.50)×10⁷/kg per infusion. Moderate fever occurred in three cases after multiple infusions, and the temperature restored to normal on the same day after treatment. Fever was observed in one patient after every infusion of four times in total. The temperature reached to 38.5-39.0 ℃ and returned to normal within 1-2 hours after adequate antipyretic treatment, and then there was no discomfort. No GVHD was observed in the elderly AML patients, while 6 cases that received allo-HSCT developed moderate acute GVHD, among them grade I in 5 cases and grade II in 1 case. No other severe toxicities were observed. CONCLUSION NK cell products with a high-purity could be obtained by ex vivo expansion with this protocol. The transfusion of these expanded cells is generally safe in the elderly patients with AML that have received chemotherapy or patients that received hematopoietic stem cell transplantation.
Aged ; Graft vs Host Disease/prevention & control* ; Hematopoietic Stem Cell Transplantation ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute/therapy* ; Middle Aged ; Recurrence

OBJECTIVE: To compare the effects of direct fluorescence in situ hybridization (D-FISH) detection without sorting and CD138 immunomagnetic bead sorting technology combined with FISH (MACS-FISH) on cytogenetic analysis of patients with multiple myeloma (MM). METHODS: FISH test results of 229 patients with initial MM were retrospectively analyzed. The patients were divided into two groups, 140 patients were tested with D-FISH and 89 patients with MACS-FISH. The combination probe was designed as P53, D13S319, RB1, 1q21, and IgH. Cytogenetic detection results were compared between the two groups. RESULTS: The total detection rate of cytogenetic abnormalities in D-FISH group was 52.9%, and that in MACS-FISH group was 79.8%. There was a significant difference in the cytogenetic abnormality rate between the two groups (P=0.020). The abnormal genes with the highest detection rate in the two groups were 1q21 and IgH, respectively, while the lowest was P53. There was no significant difference in the percentage of P53 positive cells (positive rate) between the two groups, while D13S319, RB1, 1q21, and IgH showed significant difference in positive cell rate (P=0.0002, P<0.0001, P=0.0033, P=0.0032). There was no significant correlation between the proportion of plasma cells (PC) detected by bone marrow morphology and cytogenetic abnormality rate in the D-FISH group, while there was a correlation between the proportion of PC detected by flow cytometry and cytogenetic abnormality rate (r=0.364). The PC proportion detected by bone marrow morphology and flow cytometry in the MACS-FISH group had no correlation with the cytogenetic abnormality rate and positive cell rate of the 5 genes mentioned above. Additionally, the PC proportion detected by bone marrow morphology and flow cytometry showed significant difference (P<0.0001). CONCLUSION CD138 immunomagnetic bead sorting combined with FISH technology can significantly improve the abnormality detection rate of MM cytogenetics.
Chromosome Aberrations ; Humans ; In Situ Hybridization, Fluorescence/methods* ; Multiple Myeloma/genetics* ; Retrospective Studies ; Syndecan-1/immunology* ; Tumor Suppressor Protein p53/genetics*

OBJECTIVE: To investigate the quantitative expression of immunophenotype of CD34 METHODS: Multi-parameter flow cytometry (FCM) was used to detect the proportion and mean fluorescence intensity (MFI) of each antigen of bone marrow CD34 RESULTS: Bone marrow blast cell proportion (P<0.01), RBC level (P<0.01), and Hb level (P<0.05) of high-risk MDS patients were higher, while EPO level (P<0.05) was lower than those of low-risk patients. The proportion of CD34 CONCLUSION The immunophenotype of CD34
Antigens, CD34 ; Bone Marrow ; Bone Marrow Cells ; Flow Cytometry ; Humans ; Immunophenotyping ; Myelodysplastic Syndromes

OBJECTIVE: To investigate the related factors influencing plasma transfusion efficacy so as to improve the plasma transfusion efficiency. METHODS: According to the clinical symptoms and the laboratorial results, the patients were divided into transfusion efficient and inefficient groups. A total of13090.8 units of plasma were transfused to 4423 patients. The clinical symptoms and the hemorrhage related index per- and pro-transfusion, plasma components sorts, storage time, and the dose of plasma (kg/ml) transfusion were analyzed. RESULTS: The largest transfusion volume of plasma were in intensive care unit (ICU) accounted for 30.36%, the largest blood plasma per patient transfusion was in cardiac surgery (3.96 U). The analysis of transfusion efficiency showed that in terms of patient age, there were difference in transfusion efficiency among the patients with different ages (P＜0.001). The effective transfusion rate in the group of age ＜18 was 53%, which was higher than that in group of age 18-60(41%) and group of age ＞60 (30%); in terms of sex, the effective transfusion rate in female group was higher than that in male group (42% vs 37%) (P＜0.001); in terms of transfusion plasma volume/body weight, there were differences in transfusion efficiency (P＞0.05). The multi-factor logistic regression analysis showed that there was no significant correlation among the plasma sorts, storage time of the plasma pre-transfusion and transfusion efficiency(P＞0.05). The analysis of the non-hemolytic fever reaction caused by plasma transfusion revealed that there was no statistical difference between the plasma and the leukocyte-depleted plasma groups (P＞0.05). CONCLUSION The plasma transfusion effectiveness relates with age and sex, but not relates with the transfusion plasma voume/body weight, plasma sorts, and the duration of storage.

To evaluate the right ventricular systolic function in uremia patients and the effects of maintenance hemodialysis on right ventricular systolic function by volume and strain parameters obtained by 4D RV Volume . Methods Seventy‐six patients with uremia and twenty‐two controls were selected . According to left ventricular ejection fraction ( LVEF) ,uremia patients were divided into normal LVEF uremia group and decreased LVEF uremia group . T hen normal LVEF uremia group was divided into maintenance hemodialysis group and non‐dialysis group . Conventional ultrasound parameters included :LVEF ,pulmonary artery systolic pressure ( PASP) and tricuspid annular plane systolic excursion ( T APSE‐2D) . 4D RV Volume parameters included : right ventricular end‐diastolic volume ( RVEDV ) , tricuspid annular plane systolic excursion ( T APSE‐4D ) ,right ventricular area change rate ( FAC ) ,right ventricular ejection fraction ( RVEF) and right ventricular free wall longitudinal systolic strain ( RV‐GLSfree ) . Results①Compared with the control group ,T APSE‐2D decreased significantly in the decreased LVEF uremia group ( P ＜0 .05) ,w hile there was no significant difference of T APSE‐2D in normal LVEF uremia group ( P ＞ 0 .05) . Compared with the control group and normal LVEF uremia group ,PASP increased significantly in the decreased LVEF uremia group ( P ＜ 0 .05 ) . Compared with the control group ,RVEDV increased significantly both in the normal LVEF and decreased LVEF uremia group ,w hich showed an increasing trend in these three groups ( P ＜0 .05) ,while T APSE‐4D ,FAC ,RVEF and RV‐GLSfree all decreased significantly and showed a decreasing trend in these three groups ( P ＜ 0 .05 ) . ② Compared with the control group , T APSE‐2D decreased significantly in non‐dialysis group ( P ＜0 .05) ,but there was no significant difference in uremia hemodialysis group ( P ＞0 .05) . Compared with the control group ,PASP and RVEDV increased and T APSE‐4D ,FAC ,RVEF and RV‐GLSfree decreased significantly in uremia hemodialysis group and non‐dialysis group ( P ＜ 0 .05 ) . Compared with non‐dialysis group , T APSE‐2D and T APSE‐4D increased significantly in hemodialysis group ( P ＜0 .05) ,while there was no significant difference in RVEDV ,FAC , RVEF and RV‐GLSfree in uremia hemodialysis group ( P ＞0 .05) . Conclusions 4D RV Volume could early and accurately evaluate the right ventricular systolic dysfunction in uremia patients . Furthermore ,w hen evaluating right ventricular systolic function in uremia patients treated with maintenance hemodialysis , indices such as right ventricular strain and volume parameters should be comprehensively considered .

OBJECTIVE: To investigate the effect of silencing LNK gene on the expression of EPO and EPOR in acute myeloid leukemia cells (THP-1). METHODS: THP-1 cells were cultured. The lentivirus was used as a vector to silence the LNK gene stably. After 72 hours of infection, GFP expression level was detected by the fluorescent inverted microscopy. The lentiviral Infection efficiencies were monitored by flow cytometry. The LNK silencing effect was confirmed. The mRNA expressions of EPO and EPOR were detected by RT-PCR. The protein levels of LNK, EPO and EPOR were detected by Western blot. RESULTS: At the time-point of 72 hours after lentivirus infection, the expression level of GFP was above 85% detected by fluorescent inverted microscopy. The infection efficiency was above 99% by flow cytometry. mRNA expressions of LNK, EPO and EPOR in LNK silencing group were signifycantly lower than those in control group (P＜0.05). The protein levels of LNK, EPO and EPOR in LNK silencing group were significantly lower than those in the control group (P＜0.05). CONCLUSION THP-1 cell line of LNK gene silencing has been successfully established,the LNK gene has been silenced, the expression of EPO and EPOR decrease, indicating that LNK may participate in the regulation of EPO and EPOR.
Blotting, Western ; Erythropoietin ; Gene Silencing ; Humans ; Proteins ; genetics ; Receptors, Erythropoietin ; THP-1 Cells

OBJECTIVE: To investigate the effect of LNK gene silencing and overexpression on the expression of STAT3 gene in human monocytic leukemia cells (THP-1). METHODS: THP-1 cells were cultured, and the lentivirus was used as a vector to silence and overexpres the LNK gene stably. After transfection for 72 hours, the GFP expression levels were observed by inverted fluorescence microscopy. The lentiviral transfection efficiencies were detected by flow cytometry. The effects of LNK silencing and overexpression were confirmed, and the expression of STAT3 mRNA was detected by RT-PCR. The protein levels of LNK and STAT3 were detected by Western blotting. RESULTS: The GFP expression level of THP-1 cells reached more than 85% after transfection with lentivirus for 72 hours, and the transfection efficiency of cells was above 99%. mRNA expressions levels of LNK and STAT3 in LNK silencing group were signifycantly lower than those in control group, while LNK and STAT3 mRNA levels in the LNK overexpression group was significantly higher than those in control group (P＜0.05). The protein expression levels of LNK and STAT3 in LNK silencing group were significantly lower than those in control group, while that in LNK overexpression group was significantly higher than that in control group (P＜0.05). CONCLUSION The THP-1 cell line with LNK gene silencing and overexpression has been successfully established. The LNK gene silencing resulted in decrease of STAT3 expression; LNK gene overexpression and leads to inereases of STAT3 expression indicating that LNK participates in the regulation of STAT3.
Cell Line, Tumor ; Gene Silencing ; Genetic Vectors ; Humans ; Lentivirus ; Proteins ; RNA, Small Interfering ; STAT3 Transcription Factor ; metabolism ; THP-1 Cells ; Transfection

OBJECTIVETo explore the value of of CD, MPO, Ki-67, C-MYC positive rates in the pathological tissues and C-MYC gene of patients with T-LBL/ALL for predicting Prognosis.

METHODSNinty cases of T-LBL/ALL patients in our hospital were selected and included in the T-LBL/ALL group, and 30 cases of lymphnode reactive hyperplasia were selected as control group. Immunohistochemical staining was used to detect the changes of CD, MPO, Ki-67 and C-MYC positive rate in 2 groups, and the changes of C-MYC gene were detected by fluorescence in situ hybridization.

RESULTSIn 90 patients with T-LBL/ALL, there were CD1a34 cases (37.8%), CD367 cases (74.4%), epsilon CD347 cases (52.2%), CD785 cases (94.4%), CD1033 cases (36.7%), CD3422 cases (24.4%), CD4348 cases (53.3%), CD45RO46 cases (51.1%), CD9988 cases (97.8%), TDT85 cases (94.4%); and CD23, CD20, and MPO all were negative; Ki-67>80% 47 cases (52.2% cases), Ki-67≤80%, 43 cases (47.8%). In 90 T-LBL/ALL patients, the positive rate of C-MYC (66.7%) was significantly higher than the control group (positive rate 0.0%) (P< 0.05); the Ki-67 index, mediastinal widening of T-LBL/ALL patients and the positive rate of C-MYC positively were correlated (P< 0.05). The overall survival rate (44.0%) of C-MYC negative patients was significantly higher than that of C-MYC positive patients (0.0%). The overall survival rate of C-MYC negative patients was significantly higher than that of C-MYC positive patients (P< 0.05).Ann Arbor staging, LDH, bone marrow involvement, mediastinal widening, Ki-67 positive index, and C-MYC protein expression of patients with T-LBL/ALL did not correlated with increased C-MYC gene breakage and copy number (P> 0.05).

CONCLUSIONThe overall survival rate of C-MYC positive patients decreases, which positively correlates with Ki-67 positive index and mediastinal width, suggesting that the prognosis of the patients with C-MYC protein expression is poorer.