Research Progress of Next Generation Sequencing in Acute Leukemia --Review / 中国实验血液学杂志

With the advent of precision medicine, next-generation sequencing (NGS) is playing an increasingly important role in clinical oncology diagnosis and treatment with its advantages of high sensitivity, high accuracy, high efficiency and operability. NGS reveals the genetic characteristics of acute leukemia(AL) patients by screening for specific disease-causing genes to identify occult as well as complex genetic mutations in patients with AL, leading to early diagnosis and targeted drug therapy for AL patients, as well as to predict disease recurrence by detecting mnimal residual disease (MRD) and analyzing mutated genes to determine patient prognosis. NGS plays an increasingly important role in the diagnosis, treatment and prognosis assessment in AL, providing a direction for the pursuit of precision medicine. This paper reviews the research progress of NGS in AL.

Pharmacokinetics of Recombinant Human Coagulation Factor Ⅷ Preparations in Patients with Severe Hemophilia A / 中国实验血液学杂志

OBJECTIVE@#To calculate the pharmacokinetic parameters of recombinant human coagulation factor Ⅷ using myPKFiT in patients with severe hemophilia A, and provide an individualized treatment plan for patients.@*METHODS@#A total of 42 patients with severe hemophilia A who were treated with recombinant human coagulation factor Ⅷ were included from January 2021 to December 2021. myPKFiT was used to calculate the pharmacokinetic parameters of FⅧ, and the individualized treatment plan for hemophilia A patients was formulated.@*RESULTS@#The median age of 42 patients with severe hemophilia A was 31（16-50） years old, the average weight was 54.0±9.9 kg, the half-life of FⅧ was 12.05±1.6 h, the time to more than 1% of the baseline was 62.3±15.3 h, and the 0 bleeding rate after the guidance of myPKFiT was significantly increased from 39% to 49%, the Annual bleeding rate was reduced from 3.6±2.5 to 2.1±2.0, and the Annual joint bleeding rate was reduced from 3.2±2.2 to 1.9±0.9, all of which were statistically different (P<0.05).@*CONCLUSION@#Individualized therapy in patients with severe hemophilia A who were guided by myPKFiT assay of pharmacokinetics parameters can significantly reduce the annual bleeding rate and annual joint bleeding rate of patients.

The Role of Platelet Exosomes in Atherogenic Thrombosis --Review / 中国实验血液学杂志

Exosomes are subtypes of extracellur vesicles containing a variety of cell-specific proteins, lipids and nucleic acids released during cell activation or apoptosis, and play the role of intercellur communication mediators in different physiological and pathological processes. With the development of research in recent years, the role of platelet-derived exosomes in cardiovascular diseases has attracted extensive attention. This paper reviews the role of platelet-derived exosomes in atherosclerotic thrombosis and the potential role of platelet-derived exosomes as biomarkers for the diagnosis and treatment of atherosclerotic thrombotic disease and the problems to be solved.

New Advances in the Treatment of Primary Central Nervous System Lymphoma--Review / 中国实验血液学杂志

Primary central nervous system lymphoma (PCNSL) is a rare aggressive non-Hodgkin's lymphoma outside the lymph nodes. At present, high-dose chemotherapy based on methotrexate is the standard induction therapy for newly diagnosed PCNSL, but the effective therapy of relapse/refractory and elderly PCNSL is still unclear. With the progress of clinical trials, new drugs and combined treatment method appear constantly, such as rituximab and ibrutinib, the remission rate of refractory and relapsed patients increased, while lenalidomide showed a good activity in the maintenance treatment of elderly patients. This review summarized briefly the recent advances of research on immunocheckpoint inhibitors, immunoregulatory agents, bruton tyrosine kinase (BTK) and PI3K/AKT/mTOR pathway inhibitors.

Reversal of Drug Resistance in K562/ADM Cells Caused by RA and the Related Mechanisms / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of ursane triterpenoids 3β,19α-dihydroxyursu-12-ene-23,28-dicarboxylic acid (Rotundioic acid, RA) on the sensitivity of adriamycin-resistant K562 cells (K562/ADM Cell) anti-tumor drug, and to explore the effect and mechanism of RA on the multidrug resistance of K562/ADM cells.@*METHODS@#CCK-8 method was used to detect the effect of RA on the sensitivity of K562 cells and K562/ADM cells to anti-tumor drug. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression level of mRNA and the protein in K562 and K562/ADM cells, and the effect of RA on the expression of MDR1 mRNA and P-gp in K562/ADM cells was also detected; Western blot was used to detect the expression of p-JNK, p-p38 and p-ERK1/2 in K562/ADM cells.@*RESULTS@#RA could increased the sensitivity of K562/ADM cells to adriamycin(the reversal factor was 1.61 times), the difference showed statistically significantly (P<0.05); the resistance factor of K562/ADM to ADM was 41.76 times. The expression of MDR1 mRNA in K562 cells was extremely low, and the protein product P-glycoprotein (P-gp) was almost not expressed; MDR1 mRNA and P-gp in K562/ADM cells were highly expressed; RA could down-regulate the expression levels of MDR1 and P-gp in K562/ADM cells. In addition, RA could upregulate the phosphorylation levels of p38 and ERK1/2 in K562/ADM cells, but it has no effect on the expression of p-JNK.@*CONCLUSION@#RA may participate in the regulation of MAPK signaling pathway by upregulating the expression levels of p-p38 and p-ERK1/2 in K562/ADM cells, and thus inhibit the transcription and translation levels of MDR1, and finally reverse the multidrug resistance of leukemia cells.

Expressing of linc-223 and miR-125a in Patients with Acute Leukemia and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To determine the expression level of linc-223 and miR-125a in the patients with adult acute leukemia (AL) and explore the relationship between the expression level and the occurrence, development, prognosis of leukemia.@*METHODS@#Bone marrow samples of 93 patients with AL treated in our hospital from January 2017 to September 2017 were enrolled, including 21 cases of acute lymphoblastic leukemia (ALL) and 72 cases of acute non-lymphocytic leukemia (ANLL). At the same time, bone marrow samples from 20 cases of non-malignant hematopathy patients in the same period were enrolled as control group. Real-time quantitative PCR (qRT-PCR) was used to test the expression level of linc-223 and miR-125a in bone marrow of 93 AL patients and the relationship between the level and the occurrence, development, prognosis of leukemia was analyzed.@*RESULTS@#There was no significant difference in the expression of linc-223 between AL patients (ANLL and ALL) and control group (P>0.05). Moreover, there was no significant correlation between linc-223 and PML-RARα gene or the remission rate of patients after treatment. The expression of miR-125a in ANLL patients was significantly lower than those in the control group (P0.05), and also for ALL and control group (P>0.05). In the newly treatment ANLL patients, the expression level of miR-125a showed negatively correlated with LDH level and the ratio of immature cells (r=-0.454, r=-0.400), but not with sex, degree of risk, peripheral blood leukocyte count, platelet count, hemoglobin content, WT1, CRP, etc. (P>0.05). There was a positive correlation between linc-223 and miR-125a in ANLL patients (r=0.296).@*CONCLUSION@#No abnormal expression of linc-223 was found in the bone marrow of AL patients, but miR-125a expression shows a low level and positively correlate with the expression level of linc-223 in ANLL, which is helpful for the diagnosis.

A new phenolic compound from endophytic fungus Aspergillus fumigatus of Euphorbia royleana / 中国中药杂志

This research was carried out to study the secondary metabolites of endophytic fungus Aspergillosis fumigatus from Euphorbia royleana. The endophytic fungus A. fumigatus was fermented by solid fermentation,and purified by various chromatographic methods after extraction. The structures of the compounds were identified by1 H-NMR,13 C-NMR and HSQC,HMBC spectra and physicchemical properties. Three compounds were isolated and their structures were identified as 3-( 3,4-dihydroxybenzoyl)-5-( 3,4-dihydroxyphenyl)-6-methyl-5,6-dihydro-2 H-pyran-2-one( 1),hydroxysydonic acid( 2) and 11-hydroxysydonic acid( 3). Compound 1 is a new compound.

Research Advances on the Pathogenesis of Primary Immune Thrombocytopenia--Review / 中国实验血液学杂志

Abstract　Immune thrombocytopenia (ITP) is a complex autoimmune disease characterized by less than 100×10/L platelet count in peripheral blood. The pathogenesis of ITP is complex and has not been fully elucidated. Currently, researches on the pathogenesis of ITP mainly focus on the disorders of humoral immunity and cellular immunity. In recent years, some new progress has been made in the study of this pathogenesis, including the platelet clearance pathway that is not dependent on Fc γ R mediation, the metalloproteinase (ADAM) 10 that can regulate T and B cells, and the abnormal expression of micro RNA in genetic factors. Under the joint action of multiple factors, the imbalance of the immune system in the body leads to the occurrence of ITP. This article reviews the research progress on humoral immunity, cellular immunity and other possible new pathogenesis of ITP in recent years.

Advances of Research on Application of Leukemia Xenotransplantation Model--Review / 中国实验血液学杂志

Abstract　　Tumor xenograft model (PDTX) derived from leukemia patients is an animal model in which the leukemia cells or primary cell lines of patients are transplanted directly into immunodeficient mice.The emergence of nude mice and SCID mice opened early xenotransplantation, then the NSG, NOG mice and the improved model and humanized mice based on there mice significantly improves the success rate of transplantation. The late presented transplantation of leukemia LSC and transplantation of patient-derived and induced pluripotent stem cells obtained based on iPSC technology provide new insight for the anderstanding leukemia genesis and development, and the new type humanized mouse model with normal lymphatic hematopoietic reconstruction provides a new platform of leukemia cell therapy and immunotherapy for leukemia therapy. PDTX is an important platform for the study of the pathogenesis and drug resistance mechanism of leukemia, as well as the development of new drugs and individualized treatment. In this paper, the recent progress in the construction and application of models of immunodeficient mice and their models is reviewed.

Efflect of miR-34a-Tangeted Regulation of HDACI on Apoptosis of AML Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the regulation of miR-34a on HDAC1 expression and its effect on the apoptosis of acute myeloid leukemia (AML) cells.@*METHODS@#miR-34a mimics, miR-34a inhibitor and miR-34a scramble were transfected into HL-60 cells. The effects of miR-34a expression levels on proliferation and apoptosis of HL-60 cell were detected by CCK8 assay and flow cytometry respectively. The expression of HDAC1 protein was assessed by Western blot after regulating miR-34a expression, the 3'UTR of HDAC1 was cloned and ligated to construct a dual luciferase reporter vector, and then the dual luciferase reporter assay was applied to verify the target of miR-34a, the expression vector pcDNA3.1-HDAC1 was constructed, the interaction of miR-34a and HDAC1 was analyzed by reversion test.@*RESULTS@#miR-34a over-expression could inhibit the proliferation of HL-60 cells and induce their apoptosis. Bioinformatics analysis indicated that the HDAC1 was a target gene of miR-34a. Western blot indicated that miR-34a overexpression down-regulated the expression of HDAC1. Dual luciferase reporter assay and reversion test showed that miR-34a could act at the 3-UTR of HDAC1 gene to regulate its expression.@*CONCLUSION@#miR-34a promotes the apoptosis of HL-60 cells via regulating HDAC1 expression.

Persistence of rabies antibody 3 years after postexposure prophylaxis with 2-1-1 regimen of Vero-cell rabies vaccine for human use and antibody response to two booster doses / 中华疾病控制杂志

Objective To evaluate the antibody persistence following rabies postexposure prophylaxis (PEP) with 2-1-1 regimen and antibody response to two booster doses. Methods A total of 314 healthy volunteers at year 1, year 2, year 3 who had received a complete rabies PEP using 2-1-1 regimen were recruited. Two booster doses of rabies vaccine were inoculated, and blood samples were obtained before and 14 days after two booster doses. Human rabies virus IgG antibody was evaluated by ELISA, and the antibody levels and antibody positive rates were analyzed. Results The antibody GMC of 303 people at year 1, year 2, year 3 after a complete immunization was 1.33 IU/mL, 1.04 IU/mL and 0.72 IU/mL, with an antibody positive rate of 77.78%, 66.67% and 55.56%, respectively. Among 282 people who received 2 doses for booster immunization, the antibody GMC at day 14 of 1 year, 2 year and 3 year immunization group was 16.83 IU/mL, 19.37 IU/mL and 21.05 IU/mL respectively, which was higher than that before booster immunization (t=16.54, P<0.001; t=13.85, P<0.001; t=16.02, P<0.001). The antibody positive rate was 100.00%, 99.00% and 100.00%, respectively. Conclusions The immune persistence of rabies antibody after PEP with antirabies vaccine using the 2-1-1 regimen is good so as to the immune response after 2 doses of booster immunization in 3 years is effective.

Research Progress on the Role of Musashi-2 in the Pathogenesis of Malignant Hematologic Diseases-Review / 中国实验血液学杂志

Previous studies have found that Musashi-2 (Msi-2) plays an essential role in regulating the gene expression of stem cell populations by inhibiting or activating the translation, and thus regulates stem cell function. Current research shows that Msi-2 involved in the regulation of a variety of malignant hematological diseases through different mechanisms; moreover abnormally expressed in a variety of malignant hematological diseases and involved in the occurrence, development of a variety of malignant hematological diseases. Further study on the role of malignant hematologic diseases will further clarify the pathogenesis of malignant hematologic diseases, and provide a new basis for the diagnosis and treatment of malignant hematological diseases. In this review, the structure and function of Msi-2 and its relationship with malignant hematologic diseases and its latest research progress are summarized.

Research Progress on miR-125 Family in Malignant Hematologic Diseases-Review / 中国实验血液学杂志

MicroRNAs (miRNAs) are a class of endogenous non-coding single-stranded small noncoding RNAs with the length of 20 to 23 nucleotides. MicroRNA-125 (miR-125) family, which is a highly conserved miRNA family, is consist of miR-125a, miR-125b-1 and miR-125b-2. Accumulating evidence demonstrated that miR-125 can be involved in various physiological and pathological processes in vivo. Importantly, it is closely related with the tumorigenesis and tumor development, including tumor cell proliferation, apoptosis, invasion and metastasis, metabolism and immune response. In malignant hematologic diseases, it is defined either as a oncogene, or as a tumor suppressor gene, even, closely related with the drug resistance in a variety of hematologic malignancies. MiR-125 is expected to become a new therapeutic target. Newly, the research of the relationship between miR-125 family and hematologic malignancies become increasing, including leukemia, lymphoma, multiple myeloma. In this review, the relationship between miR-125 family with malignant hematologic diseases and its latest research progress are summarized.

Research Progress on Role of Inflammasome in Development of Thrombotic Diseases -Review / 中国实验血液学杂志

Inflammasome is a group of polyprotein complexes located in the cytoplasm, its activation can induce the maturation and release of proinflammatory cytokines IL-1β and IL-18, and promote the early atherosclerosis. In the recent years, it is found that the inflammasome is activated in thrombotic deseases, moveover, the activated inflammasome and its activation induced cytokines promote the occurrence and development of thrombolic deseases, and show the unfavaourable effect on prognosis. With further exploration on the mechanisms of thrombotic diseases, the relationship between the inflammasome and thrombotic diseases increasingly become a hot spot of research. This review focuses on the action mechanisms of inflammasome in thrombotic diseases.

Research Progress on Application of CRISPR/Cas Genome Editing Technology in Hematological Diseases -Review / 中国实验血液学杂志

CRISPR/Cas genome editing technology is a newly developed powerful tool for genetic manipulation, which can be used to manipulate the genome at specific locations precisely, to restore the function of genetic defect cells, and to develop various disease models. In recentl years, with the advances of precise genome manipulation, CRISPR/Cas technology has been applied to many aspects of diseases research and becomes an unique tool to investigate gene function and discover new therapeutic targets for genetic diseases. Nowadays, CRISPR/Cas technology has been a hot research point in agriculture, graziery, biotechnology and medicine. This review focuses on the recent advances in CRISPR/Cas technology and its application in hematological diseases.

Expression and Significance of NLRP3, ASC and AIM2 in Patients with Acute Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the expression and significance of NLR family, pyrin domain containing 3 (NLRP3), apoptosis associated speck like protein containing a CRAD (ASC) and absent in melanoma 2 (AIM2) of patients with acute leukemia.</p><p><b>METHODS</b>The petipheral blood samples of 19 patients with ALL and 41 patients with ANLL as the AL group (each 20 cases of newly diagnosed, relapsed and complete remission group) and 20 cases of non-hematologic malignancies as the control group were collected from July 2013 to July 2014 in the First Affiliated Hospital of Gannan Medical University. The expression levels of NLRP3, ASC and AIM2 in peripheral blood plasma were determined by ELISA.</p><p><b>RESULTS</b>The expression levels of NLRP3, ASC and AIM2 in plasma of control and AL complete remission groups were significantly higher than those in newly diagnosed and relapsed groups, and were with statistical significance (P < 0.05), but there were no statistical signifirance between ALL and ANLL groups (P > 0.05).</p><p><b>CONCLUSION</b>The expression of NLRP3, ASC and AIM2 is down-regulated in the patients with acute leukemia, which maybe play a role of anti-leukemia, and provide a laboratory evidence for diagnosis and treatment of patients with acute leukemia.</p>

Research Progress on Long Non-Coding RNA in Hematological Malignancies--Review / 中国实验血液学杂志

Long non-coding RNA (LncRNA) is defined as a class of transcripts more than 200 nucleotides in length and without the protein-coding function. It has been found for years, however, that little is known about the potential role of LncRNA in humans. But recent studies showed that LncRNA can regulate the coding-gene expression and participate in effects of human body. Accumulating evidence demonstrated that LncRNA are involved in cancer incidence, development and progression.With further exploration on the mechanisms of tumors, the relationship between the long non-coding RNA and hematological malignancies increasingly become a hot research. This review focuses on the mechanisms of LncRNA in hematological malignancies.

Expression of HMGB1 and NF-κB in Patients with Acute Leukemia and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the expression of high mobility group box protein 1 (HMGB1) and nuclear factor-kappa B (NF-κB) in patients with acute leukemia and its significance.</p><p><b>METHOD</b>20 samples of bone marrow and peripheral blood from each acute leukemia groups (newly diagnozed, relapsed and complete remission groups) and 20 samples as control from patients with no-hematologic malignancies were collected. The expression level of HMGB1 in peripheral blood plasma was determined by ELISA; HMGB1 and NF-κB level in mononuclear cells were examined by RT-PCR. Western blot was used to determine HMGB1 and NF-κB protein levels. HMGB1 and NF-κB in bone marrow smears were determined by immnohistochemistry method (IHC).</p><p><b>RESULTS</b>The expression level of HMGB1 obviously increased in patients of newly diagnosed and relapsed groups, as compared with control group there was statistical significance (P < 0.05), but there was no obvious difference in expression level of HMGB1 between complete remission group and control group (P > 0.05). The expression level of HMGB1 and NF-kB in monnuclear cells of bone marrow in newly-diagnosed group and relapsed group was significantly higher than that in control group (P < 0.05), but the expression levels of HMGB1 and NF-kB in complete remisson group did not change (P > 0.05). The results of immnohistochemistry method indicated that the possitive expression of HMGB1 and NF-kB maily was found in bone marrow smears of newly diagnosed and relapsed groups.</p><p><b>CONCLUSION</b>HMGB1 is overexpressed in acute leukemia, which may be involved in the occurrence and development of acute leukemia by activating the NF-κB signaling pathway, HMGB1 may be a important index for observing therapeutic effectiveness and predicting recurrence of acute leukemia.</p>

Clinical study of 32 patients with adult Philadelphia chromosome-positive acute lymphoblastic leukemia / 中国实验血液学杂志

This study was aimed to evaluate the efficacy and safety of imatinib in the treatment of patients with adult Ph chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL). A total of 32 diagnosed adult Ph(+)ALL patients from July 2007 to February 2014 in our hospital were retrospectively analyzed and were divided into two groups: imatinib plus chemotherapy group and traditional chemotherapy group. The differences between two groups were analysed in disease-free survival time (DFS), overall survival time (OS) and toxicity. The G banding technigue was used to analyse the karyotype, and the flow cytometry was applyed to detect the immune markers on surface of cells. The results showed that all patients expressed B cell and hematopietic stem/progenitor cell immune markers, out of them 21 patients (65.6%) were with myeloid antigens, 27 patients with simple Ph (+) phenotype and 5 patients with additional chromosome abnormality. The DFS and OS of the imatinib group were statistically longer than those of the traditional chemotherapy group (14.3 ± 4.7 months vs 10.7 ± 3.8 months) (P < 0.05) and 22.6 ± 6.8 months vs 10.7 ± 3.8 months) (P < 0.05)). There was no significant difference in toxic effects between two groups (P > 0.05)). It is concluded that the all cases of adult Ph(+)ALL are with B cell phenotype and express hematopietic stem/progenitor cell antigen. They often accompanied by expression of myeloid antigens and additonal chromosome abnormality in genetics. The combination of imatinib with chemotherapy can prolong remission time and survival time for patients of non-hematopietic stem cell transplantation on the basis of no notably increasing the toxic effects.

Apoptosis of Burkitt's lymphoma Raji cell line induced by bortezomib / 中国实验血液学杂志

The aim of this study was to clarify whether bortezomib might induce apoptosis in Burkitt's lymphoma Raji cell line and its mechanism. Different concentrations of bortezomib were used to treat Raji cells and its effects of time and dose were observed. Cell morphology was observed under light microscope; flow cytometry was used to analyze cell apoptosis; RT-PCR was used to detect the expressions of NF-kappaB and p53 gene mRNAs. The results showed that the bortezomib could inhibit Raji cell growth within a certain range of treating time and dose. Apoptosis were induced in relation to time and dose. The expression of NF-kappaB mRNA and p53 mRNA decreased after treatment with bortezomib. It is concluded that the bortezomib can induce Raji cell apoptosis, which provides a theoretical basis for clinical treatment. NF-kappaB and p53 gene are supposed to participate in the bortezomib induced apoptosis of Raji cells.