OBJECTIVE: To explore the application of targeted mRNA sequencing in fusion gene diagnosis of hematologic diseases. METHODS: Bone marrow or peripheral blood samples of 105 patients with abnormally elevated eosinophil proportions and 291 acute leukemia patients from January 2015 to June 2023 in the First Affiliated Hospital of Soochow University were analyzed and gene structural variants were detected by targeted mRNA sequencing. RESULTS: Among 105 patients with abnormally elevated eosinophil proportions, 6 cases were detected with gene structural variants, among which fusion gene testing results in 5 cases could serve as diagnostic indicators for myeloid neoplasms with eosinophilia. In addition, a IL3∷ETV6 fusion gene was detected in one patient with chronic eosinophilic leukemia, not otherwise specified. Among 119 patients with acute myeloid leukemia (AML), 38 cases were detected structural variants by targeted mRNA sequencing, accounting for 31.9%, which was significantly higher than 20.2% (24/119) detected by multiple quantitative PCR (P < 0.05). We also found one patient with AML had both NUP98∷PRRX2 and KCTD5∷JAK2 fusion genes. A total of 104 patients were detected structural variants by targeted mRNA sequencing in 172 cases with acute B-lymphoblastic leukemia who were tested negative by multiple quantitative PCR, with a detection rate of 60.5% (102/172). CONCLUSION Targeted mRNA sequencing can effectively detect fusion gene and has potential clinical application value in diagnosis and classificatation in hematologic diseases.
Humans ; Hematologic Diseases/diagnosis* ; RNA, Messenger/genetics* ; Oncogene Proteins, Fusion/genetics* ; Sequence Analysis, RNA ; Leukemia, Myeloid, Acute/diagnosis*

Humans ; Hearing Loss/etiology* ; Lupus Erythematosus, Systemic/complications* ; Prognosis

Objective:To explore the effect of case three-dimensional teaching method (CTTM) combined with multimedia teaching on the practice teaching of liver infectious diseases.Methods:Twenty-five interns of liver infectious diseases received by The Second Hospital of Nanjing from May 2019 to January 2020 were taken as the control group, and 25 interns of liver infectious diseases received from February 2020 to October 2020 were taken as the observation group. The control group was given the conventional teaching mode including the conventional case teaching and the conventional multimedia teaching, while the observation group was given CTTM integrated with multimedia teaching method, and both groups were taught for 4 weeks. After teaching, the examination results of liver infectious disease related theory and practice, the changes of core competence before and after teaching, and the recognition of teaching were compared between the two groups. SPSS 25.0 was used for t test and chi-square test. Results:After teaching, the examination results of liver infectious disease related theory [(92.45±7.23) vs. (81.21±7.11)] and practice [(93.27±6.68) vs. (81.23±6.26)] in the observation group were higher than those in the control group. There was no significant difference in the scores of core competence between the two groups before teaching; after teaching, the scores of autonomous learning, problem analysis and solving, doctor-patient communication and teamwork ability of the two groups were higher than those before teaching, and the scores of observation group were significantly higher than those of the control group ( P<0.05). The recognition rates of interns in the observation group were higher than those in the control group in the improvement of learning initiative, the consolidation of theoretical knowledge, the enhancement of practical ability and the improvement of learning efficiency ( P<0.05). Conclusion:In the teaching of liver infectious disease interns, CTTM integrated with multimedia teaching method can not only improve the examination results, and enhance the core competence, but also improve the recognition of interns on the teaching mode.

OBJECTIVE: To quantitatively evaluate the non-carcinogenic and carcinogenic risks of low-level dichloromethane(DCM) exposure. METHODS: A typical sampling method was used to select workers from an engine factory and an automobile parts factory as the research subjects. After hazard identification and hazard characteristic assessment, the occupational health risks of DCM in the workplace were quantitatively assessed by non-carcinogenic risk assessment and carcinogenic risk assessment. RESULTS: The highest exposure concentration of short-term of DCM was 43.6 mg/m~3 in the air of eight DCM job posts in these two enterprises, which was lower than the operational level of 300.0 mg/m~3(half of the occupational exposure limit in China). Under this premise, the non-carcinogenic risk assessment results showed that the non-carcinogenic risk caused by inhalation of DCM was unacceptable in workers of these eight job posts. The results of carcinogenic risk assessment showed that the carcinogenic risk caused by inhalation of DCM was acceptable for all the workers in these eight job posts. CONCLUSION: Low-level DCM exposure can cause unacceptable occupational health risks to workers. Quantitative risk assessment can be applied to assess exposure to low-level chemicals.

OBJECTIVE: To analyze the effect of clinical features, routine laboratory examination and related gene mutation on the OS of patients with myelodysplastic syndrome (MDS) after hematopoietic stem cell transplantation (HSCT). METHODS: 121 patients diagnosed as MDS and underwent hematopoietic stem cell transplantation in the First Affiliated Hospital of Soochow University from October 2013 to August 2018 were selected. Basic information of the patients was collected, and blood cells, bone marrow blasts at initial diagnosis, chromosomal karyotypes and gene mutations of the patients were detected.The effect of different factors on overall survival (OS) was analyzed by statistical method. RESULTS: Kaplan-Meier univariate analysis shows that OS was significanly different among different age groups. The 3-year OS rate of patients aged 0-29 years was (83.3±7.7) %, the 3-year OS rate in patients aged 30-49 years was (58.1±7.7 %), and the 3-year OS rate of patients aged 50-69 years was (31.0±22.6) %, which was statistically different (P＜0.05) between different groups. There were also significant differences in OS among patients with different transplantation types. 3-year OS rate: HLA-matched sibling HSCT＞unrelated HLA-matched HSCT＞haploidentical HSCT＞micro HSCT. The OS rate of patients with bone marrow blasts≥10% seems lower than blasts＜10%, but there was no statistical difference.The 3-year OS rate of patients with chromosomal karyotype complex abnormality was (47.7±11.5) %, and that of patients without complex abnormality was (80±4.2) % which was statistical difference (P＜0.05). Patients with DNMT3A, NRAS, TP53 and GATA2 mutations had shorter OS time compared with patients without mutation of these genes, which shows statistically significant (P＜0.05). COX multivariate analysis showed that age, chromosome karyotype, DNMT3A, TET2, GATA2 and NRAS were the independent factors influencing OS of patients after HSCT, with statistically significant difference. CONCLUSION age of patients, donor selection of HSCT, chromosome karyotype, DNMT3A, NRAS, TP53, GATA2 and TET2 gene mutations are all independent factors affecting the OS of patients after HSCT. Therefore, the assessment of the OS of MDS patients with transplantation requires comprehensive consideration.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Infant ; Infant, Newborn ; Middle Aged ; Myelodysplastic Syndromes ; Prognosis ; Retrospective Studies ; Siblings ; Survival Analysis ; Young Adult

OBJECTIVETo explore the role of bromodomain and extra terminal (BET) bromodomain in hematopoietic differentiation from human enbryonic stem cells (hESC).

METHODSThe effect of BET hematopoietic inhibitor I-BET151 on hematopoietic differentiation from hESC was detected by using a monolayer hematopoietic defferentiation model, immunofluorescence, flow cytometry and real-time PCR; moreover the role of I-BET151 in process of hematopoietic differentiation was explored by adding I-BET151 in different differentiation stages.

RESULTSThe analysis results of immunofluorescence, flow cytometry and real-time PCR showed that I-BET 151 significantly inhibited the generation of CD43 positive hematopoietic stem and progenitor cells (HSPCs). It was found that the addition of I-BET 151 in different stages, including APLNR lateral plate mesoderm production, CD34CD31 hemogenic endothelium (HEP) generation and endothelial-to-hematopoietic transition, significantly suppressed the generation of CD43 positive hematopoietic progenitor cells.

CONCLUSIONI-BET 151 inhibites hematopoietic differentiation from hESCs at several stages, suggesting that the BET bromodomain plays important roles in multiple stages of hematopoietic differentiation from hESCs.

Apelin Receptors ; Cell Differentiation ; Flow Cytometry ; Hemangioblasts ; Hematopoietic Stem Cells ; Human Embryonic Stem Cells ; Humans

OBJECTIVETo explore the differentiation-inducing potentiality of Pulsatilla saponin A on K562 cells.

METHODSPulsatilla saponin A of different concentrations was used to treat K562 cells; the benzidine staining and the hemoglobinometry were applied to measure the change of hemoglobin content; the flow cytometry (FCM) was used to detect the expression of CD71 and GPA on K562 cells.

RESULTSK562 cells treated with 4 µg/ml pulsatilla saponin A differentiated into the erythroid lineage. With the treatment of pulsatilla saponin A, the hemoglobin content in K562 cells increased significantly; CD71 and GPA expression on the K562 cell surface were up-regulated.

CONCLUSIONPulsatilla saponin A can induce K562 cells to differentiate into erythroid lineage.

Antineoplastic Agents ; Cell Differentiation ; Cell Lineage ; Erythroid Cells ; Humans ; K562 Cells ; Saponins

OBJECTIVETo explore the autophagy activity of CD34+ cells in bone marrow of MDS patients and its clinical significance.

METHODSThe activity of autophagy in bone marrow CD34+ cells from 20 MDS patients, 20 non-malignant anemia patients and 5 AML patients admitted in our hospital from October 2012 to March 2014 was detected by flow cytometry (FCM).

RESULTSThe autophagy activity in low risk MDS patients and non-malignant anemia patients were both significantly higher than that in both high risk MDS and AML patients (P<0.05), and more interestingly, the autophagy activity in MDS negatively correlated with World Health Organization classification-based prognostic system (WPSS) score (r=-0.877) .

CONCLUSIONThe autophagy activity CD34+ cells in the patients with MDS is higher than that in AML patients, and negatively correlated with WPSS scores, indicating that the decrease of autophagy activity maybe accelerate the genesis and development of MDS and relate with the prognosis of MDS patients.

Antigens, CD34 ; metabolism ; Autophagy ; Bone Marrow Cells ; cytology ; pathology ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; pathology ; Myelodysplastic Syndromes ; pathology ; Prognosis

OBJECTIVETo summarize the clinical characteristics as well as diagnosis and treatment in 1 case of acute myeloid leukemia(AML) with coexpression of Ph and inv(16).

METHODSA series of clinical tests, the cellular morphological, immunological, cytogenetic and molecular biological examinations of leukemia cells were performed.

RESULTSThe clinical characteristics of this patient were very common. The cellular morphology is similar to the AML with inv(16). The leukemia cells were stained positively for CD13, CD33, CD34, CD117 and HLA-DR. Karyotypic analysis showed a complex chromosome abnormality including inv(16) and Ph, and the FISH analysis showed that the percentage of rearrangement of CBFβ allele was over that of the BCR-ABL fusion signals. The obvious adverse events did not occur in this patient within 3 years.

CONCLUSIONPh as secondary aberration of inv(16) rarely occures in primary AML cases, and so far there have not been the clear criteria of diagnosis and treatment. The cytogenetic and molecular biology could provide the basis for diagnosis. Moreover, autologous hematopoietic stem cell transplantation combined with imatinib probably is one of the effective treatment methods.

Chromosome Aberrations ; Chromosome Disorders ; Chromosome Inversion ; Fusion Proteins, bcr-abl ; HLA-DR Antigens ; Humans ; Leukemia, Myeloid, Acute ; Philadelphia Chromosome

Most Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL) patients often show rapid recurrence and development of ABL kinase domain (KD) mutation after tyrosine kinase inhibitor (TKI) treatment. To further investigate the mechanism of Ph(+) ALL fast relapse after TKI treatment, ABL KD mutation in 35 Chinese Ph(+) ALL with TKI resistance was detected by direct sequencing. The results showed that 77.1% (27/35) Ph(+) ALL patients with TKI resistance had ABL KD mutation and 55.6% (15/27) Ph(+) ALL patients with ABL KD mutation had T315I. Interestingly, 77.8% (21/27) Ph(+)ALL showed ABL mutation G: C→A:T, including T315I, E255K and E459K. Furthermore, all the Ph(+) ALL patients with two or more ABL KD mutations collaborated with complex chromosome abnormality and all the TKI-resistant Ph(+) ALL patients, whose karyotype progressed from simple t (9;22) into complex, developed ABL KD mutation. Moreover, the expression level of uracil-DNA glycosylase UNG2, which inhibits G:C→A:T transition in genomic DNA, decreased in Ph(+) ALL with TKI-resistance compared to that in newly diagnosis Ph(+) ALL. It is concluded that there is a high frequent ABL KD G:C→A:T mutation and a high genomic instability in Chinese TKI-resistant Ph(+) ALL. In addition, the decreased UNG2 expression in TKI-resistant Ph(+) ALL probably contributes to their high rate of ABL KD G:C→A:T mutation.
Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; DNA Glycosylases ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Male ; Middle Aged ; Point Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Protein Kinase Inhibitors ; pharmacology ; Uracil-DNA Glycosidase ; genetics