Objective:To investigate the characteristics of lactate dehydrogenase A (LDHA) gene expression in bone marrow mononuclear cells of children with acute myeloid leukemia (AML), and its association with clinical efficacy and prognosis, and the clinical significances.Methods:A retrospective case series study was conducted. Forty newly-treated AML (non-acute promyelocytic leukemia) children confirmed by morphology, immunology, cytogenetics, and molecular biology at the Children's Hospital Affiliated to Zhengzhou University from February 2021 to March 2022 were included in the AML group, and the pediatric patients were given induction therapy, consolidation therapy and maintenance therapy according to the Chinese Children's Leukemia Group AML-2019 protocol. Eighteen children undergoing bone marrow examination for non-hematologic diseases (iron-deficiency anemia, idiopathic thrombocytopenic purpura, etc.) during the same period served as the control group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect LDHA mRNA expression levels in bone marrow mononuclear cells before treatment. Patients were categorized into high and low LDHA expression groups based on the median relative expression of LDHA mRNA. LDHA mRNA levels were compared between the AML and control groups, as well as among AML patients with different treatment responses. The distribution of LDHA high or low expression patients was analyzed across clinical characteristic subgroups. Kaplan-Meier method was used for overall survival analysis of children with high and low expression of LDHA, and log-rank test was used for inter group comparison.Results:There were 26 males (65.0%) and 14 females (35.0%) in the AML group, with an age of (4.9±3.3) years; there were 9 males (50.0%) and 9 females (50.0%) in the control group, with an age of (4.2±2.8) years; there was no statistically significant difference in age and gender between the two groups (both P > 0.05). The relative expression level of LDHA mRNA before treatment in bone marrow mononuclear cells of AML patients was higher than that of the control group (4.32±1.21 vs. 1.05±0.38), and the relative expression level of LDHA mRNA before treatment in AML patients who did not improve after induction therapy (8 cases) was higher than that in AML patients who achieved complete remission (CR) after induction therapy (32 cases) (5.89±1.45 vs. 3.76±1.02), and the differences were statistically significant (both P < 0.05). The median relative expression level of LDHA mRNA in AML patients was 2.90, and 20 patients with high LDHA expression and 20 patients with low LDHA expression were classified according to this value. The proportion of AML patients with LDHA high expression in the CR after induction therapy group was lower than that in the NR group [37.5% (12/32) vs. 8/8], and the difference was statistically significant ( P = 0.003). However, there was no statistically significant difference in the proportion of patients with high LDHA expression among the subgroups of gender, age ≥ 10 years, France-America-Britain classification, white blood cell count ≥ 50×10 9, and risk stratification (all P > 0.05). Forty AML children were followed up for a median of 23 months (ranging from 6 to 36 months); 9 cases (22.5%) died, including 2 cases with LDHA low expression and 7 cases with LDHA high expression, accounting for 10.0% and 35.0% of the two groups, respectively. Kaplan-Meier survival analysis showed that the overall survival of the LDHA low expression group was better than that of the LDHA high expression group, and the difference was statistically significant ( P = 0.036). Conclusions:LDHA is highly expressed in bone marrow mononuclear cells of children with AML, and its expression level may be related to the efficacy and overall survival, which may be a potential biomarker for prognostic evaluation.

Objective:To summarize the morphological characteristics of diffuse large B-cell lymphoma (DLBCL) cells and investigate the prognosis value of the characteristics and the number of DLBCL cells in bone marrow.Methods:Retrospective study. We collected 79 cases newly diagnosed with DLBCL in the First Affiliated Hospital of Zhengzhou University from January 2020 to August 2022. 30 cases newly diagnosed without bone marrow involvement were selected as controls, whose mean age 58 years (30-82 years). The DLBCL cells were evaluated by the bone marrow smear, biopsy and flow cytometry separetely.The detection rate of DLBCL cells in the bone marrow was compared, to analyse the relationship between the morphological characteristics of DLBCL in the smear, clinical characteristics and flow cytometry parameters, and the prognostic value of DLBCL detected in the bone marrow smear and its quantity was analyzed. Logistic regression was used to analyze the correlation between the detection of DLBCL cells in bone marrow smears and the age, clinical stage, and the number of extraderules involved organs. Multivariate Cox regression was used to analyze the influence of DLBCL cells detection and its number on the prognosis of patients.Results:(1) The positive rates of DLBCL cells in bone marrow biopsy, bone marrow smear and flow cytometry were 4.86%, 5.14% and 9.27% respectively. (2) The morphological characteristics of 79 cases in bone marrow smears were described: more than 2 times the volume of the cell body of the lymphocyte, the shape was different, round or quasi-round or irregular shape, can be seen pseudopodia or protrusion; The volume of cytoplasm was moderate, vacuoles were visible, and a few perinuclear areas were visible. The nucleus were different in shape, round or quasi-round or irregularly shaped, with a majority of them having multiple nuclei and a few of them having delicate and loose chromatin. Most nucleoli were medium or large obviously, with a majority of them having 1-2 nucleoli and a few having more than 3.Sergiosomes and hemophagocytosis were observed in some DLBCL cases, tumor cell aggregation phenomenon was observed in a few DLBCL cases, occasionally pathological mitosis.(3) DLBCL cells in bone marrow smear was positively related to the age of patients, clinical stage and the number of extranodal organs involved(regression coefficient were 2.012, 2.754, 2.028, P<0.05);The volume of DLBCL cells in bone marrow smear was positively correlated with the ratio of CD4 and CD8(regression coefficient is 2.545, P<0.05);The vacuoles in cytoplasm and the pseudopod of tumor were both negative relationship with the quantity of CD38 expressed on DLBCL cells(regression coefficient was -2.465, -3.045, P<0.05); (4) DLBCL cells in bone marrow smear was an independent risk factor for PFS and OS( RR=7.059, P<0.05); RR=5.409, P<0.05). Conclusion:The appearace of DLBCL cells in bone marrow smear with prognosis, and could be used for clinical staging.

Objective:To investigate the correlation between the number of circulating DLBCL cell and the marrow tumor cell burden and the prognostic indicators in patients with DLBCL, and to evaluate the feasibility of circulating DLBCL cell reflecting the marrow tumor burden and disease progression. Optimization of FCM for screening circulating DLBCL cell was done to monitor MRD and recurrence.Methods:We conducted a retrospective study in 75 diagnosed DLBCL patients in the First Affiliated Hospital of Zhengzhou University from June 2020 to February 2021, including 43 males and 32 females aged 61 (37-85) years. According to the diagnosis and treatment criteria, the patients were divided into initial and recurrence group ( n = 53), partial response(PR)group ( n=14) and complete response(CR)group ( n=8). According to the positive criteria of circulating DLBCL cells, 48 cases were divided into circulating DLBCL positive group and 27 cases were negative group. 30 anemia patients with non-B-cell tumor-related diseases were selected as the control group, including 16 males and 14 females, aged 52 (30-79) years. 70 healthy subjects, including 36 males and 34 females, aged 39 (25-57), were selected for methodology optimization. FCM was used to detect the ratio of marrow and circulating DLBCL cells in each group, and analyze the connection between circulating DLBCL cells and clinical indicators. Statistical analysis was performed using t test, χ 2 test, Kruskal-Wallis H test, Spearman rank correlation, and Logistic regression. Results:(1) Bone marrow and circulating DLBCL cells were not detected in CR group and control group; The positive rate of circulating DLBCL cells in the initial/recurrent group and PR groups was 75.47% and 57.14%, respectively. The proportion of bone marrow and circulating DLBCL cells was positively correlated in the two groups ( P value was <0.001 and 0.020, respectively). (2) The proportion of bone marrow and circulating DLBCL cells in the initial and recurrent groups, PR group, CR group and control group decreased successively ( P<0.05). The proportion of DLBCL cells was 27.72% (initial and recurrent bone marrow group), 26.92% (initial and recurrent circulating group), 3.23% (bone marrow PR group) and 1.67% (circulating PR group), respectively. (3) Compared with the negative group, the circulating DLBCL cell positive group had increased LDH, β 2-MG, and CMYC expression(≥80%), with decreased LYM, HGB<100 g/L, B symptoms, PD-L1 expression, and age ≥60 years, showing higher ECOG, aaIPI/IPI scores and Ann staging ( P<0.05). Age ≥60, B symptoms, and PD-L1 expression were independent risk factors for circulating DLBCL cells ( P<0.05). Conclusions:The detectable rate of circulating DLBCL cell could be improved by optimizing the preoperative treatment conditions of FCM. Circulating DLBCL cells can reflect the tumor burden and disease progression. Detecting circulating DLBCL cells may improve patients′ compliance.

Idiopathic thrombocytopenic purpura (ITP) is a common bloody disease with a high incidence in children, but its diagnostic method is exclusive diagnosis, and the existing detection techniques are mostly invasive, which may cause secondary injury to patients and also may increase the risk of disease. In order to make up for the lack of the detection method, this study made a preliminary exploration on the diagnosis of children's ITP from the perspective of infrared thermography. In this study, a total of 11 healthy children and 22 ITP children's frontal infrared thermal images were collected, and the pattern characteristic (PFD), average temperature (Troi) and maximum temperature (MAX) characteristics of 7 target areas were extracted. The weighted PFD parameters were correlated with the platelet count commonly used in clinical diagnosis, and the sensitivity and specificity of the weighted PFD parameters for children's ITP were calculated through the receiver operating characteristic curve (ROC). The final results showed that the difference of the weighted PFD parameters between healthy children and ITP children was statistically significant, and the parameters negatively correlated with platelet count. Under the ROC curve, the area under the curve (AUC) of this parameter is as high as 92.1%. Based on the research results of this paper, infrared thermography can clearly show the difference between ITP children and healthy children. It is hoped that the methods proposed in this paper can non-invasively and objectively describe the characteristics of ITP infrared thermal imaging of children, and provide a new ideas for ITP diagnosis.
Area Under Curve ; Child ; Humans ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; Thermography

Objective To setup a measurement of human bone marrow micromegakaryocyte which based on CD41a and PI double‐labeled flow cytometric analysis ,and study the significance in the diagnosis of MDS .Methods In 42 cases of MDS patients , their bone marrow megakaryocytes were obtained by Percoll density gradient separation medium .The megakaryocyte glycoproteinⅡb/Ⅲa(CD41a)were marked with fluorescein isothiocyanate through its corresponding monoclonal antibody ,and their DNA were marked with PI .Then the megakaryocyte ploidy was analyzed by flow cytometry(FCM ) .Results The method for micromegakaryo‐cyte identification and analysis was established .In 42 patients with MDS ,the detection rate of micromegakaryocyte was 90 .5 per‐cent by FCM analysis ,but only 54 .8 percent by Wright‐Giemsa staining test and 64 .3 percent by immunohistochemistry ,the differ‐ence among them was statistically significant(χ2 = 13 .640 ,P= 0 .001) .The 42 patients with MDS were divided into two groups (low‐risk group and high‐risk group) .The detection rates of micromegakaryocyte were 81 .8 percent in low‐risk group and 100 per‐cent in high‐risk group separately by FCM analysis ,the difference was statistically significant(χ2 =4 .019 ,P=0 .045) .Conclusion The detection rate of micromegakaryocyte by FCM with CD41a and PI double marker is higher than that by cytochemical staining . The detection rate of micromegakaryocyte in the high‐risk group is higher than that of the low‐risk group ,which shows that the de‐tection of micromegakaryocyte is of great significance for MDS prognosis assessment .

Dendritic Cells ; pathology ; Hematologic Neoplasms ; pathology ; Humans ; Skin

We report two cases of rituximab (RTX)-induced interstitial pneumonia in two lymphoma patients receiving RTX treatment. Interstitial pneumonia was successfully managed in these two cases after a one-week-long intervention with corresponding treatments without affecting further treatment of the primary disease. RTX-induced interstitial pneumonia is characterized by a latent onset with an unclear pathological mechanism and absence of typical symptoms. High-resolution CT scan can provide valuable evidence for early diagnosis of RTX-induced interstitial pneumonia, which might be attributed partially to an increased susceptibility to P. jirovecii and fungal infection due to prolonged RTX treatment.

We report two cases of rituximab (RTX)-induced interstitial pneumonia in two lymphoma patients receiving RTX treatment. Interstitial pneumonia was successfully managed in these two cases after a one-week-long intervention with corresponding treatments without affecting further treatment of the primary disease. RTX-induced interstitial pneumonia is characterized by a latent onset with an unclear pathological mechanism and absence of typical symptoms. High-resolution CT scan can provide valuable evidence for early diagnosis of RTX-induced interstitial pneumonia, which might be attributed partially to an increased susceptibility to P. jirovecii and fungal infection due to prolonged RTX treatment.

Objective To investigate the effect of simulated microgravity on the proliferation and differentiation of the human megakaryocyte cells in vitro. Methods The fourth generation rotating cell culture system (RCCS-4) was used to generate the simulated microgravity environment. The cell viability was assessed by trypan blue staining method. The proliferation of cells was assessed by cell counting method and CCK8 method. The CD41+/CD61+ cells rate and the cells cycle were detected by flow cytometry. The expression levels of thrombopoietin receptor (c-mpl) and transcription factors were detected with RT-PCR. Results After 24, 48, 72 h, culture under simulated microgravity resulted in a significant decrease in the cell number , proliferative activity, cells in the G2/M phase and levels of c-mpl mRNA expression in comparison with that under the normal gravity (P < 0.05). After 48 h and 72 h culture, CD41+/CD61+ cells ratio decreased and RUNX-1 mRNA expression was down-regulated in cells of the group SMG compared with that of the group NG (P < 0.05). Conclusion Microgravity can inhibit the proliferation and differentiation of human megakaryocyte cells in vitro. The mechanism may be that TPO/c-mpl pathway was inhibited by down regulating the expression of c-mpl which transcriptional inhibition lead to.