Treatment response of a two-dose regimen of dose-adjusted inotuzumab ozogamicin in relapsed/refractory B-cell acute lymphoblastic leukemia / 中华血液学杂志

Objective: To observe the treatment response of a two-dose regimen of inotuzumab ozogamicin (inotuzumab), a monoclonal antibody targeting CD22, for patients with heavily treated relapsed/refractory B-cell acute lymphoblastic leukemia (R/R B-ALL), including those failed or relapsed after chimeric antigen receptor (CAR) -T-cell therapy. Methods: Pediatric and adult patients who received two doses of inotuzumab and who were evaluated after inotuzumab treatment were included. Antibody infusions were performed between March 2020 and September 2022. All patients expressed CD22 antigen as detected by flow cytometry (>80% leukemic cells displaying CD22) before treatment. For adults, the maximum dosage per administration was 1 mg (with a total of two administrations). For children, the maximum dosage per administration was 0.85 mg/m(2) (no more than 1 mg/dose; total of two administrations). The total dosage administered to each patient was less than the standard dosage of 1.8 mg/m(2). Results: Twenty-one patients with R/R B-ALL were included, including five children (<18 years old) and sixteen adults. Seventeen patients presented with 5.0% -99.0% leukemic blasts in the bone marrow/peripheral blood or with extramedullary disease, and four patients were minimal residual disease (MRD) -positive. Fourteen patients underwent both CD19 and CD22 CAR-T-cell therapy, four underwent CD19 CAR-T-cell therapy, and three underwent blinatumomab therapy. Eleven patients underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). After inotuzumab treatment, 14 of 21 patients (66.7% ) achieved a complete response (CR, one was MRD-positive CR), and all four MRD-positive patients turned MRD-negative. Four of six patients who failed recent CD22 CAR-T-cell therapy achieved a CR after subsequent inotuzumab treatment. Seven patients (33.3% ) demonstrated no response. Grade 1-3 hepatotoxicity occurred in five patients (23.8% ), one child with no response experienced hepatic veno-occlusive disease (HVOD) during salvage transplantation and recovered completely. Conclusion: For patients with heavily treated R/R B-ALL, including those who had undergone allo-HSCT and CD19/CD22 CAR-T-cell therapy, the two-dose regimen of inotuzumab resulted in a CR rate of 66.7%, and the frequency of hepatotoxicity and HVOD was low.

Prognostic significance of Notch1 protein expression in children with pediatric B-cell acute lymphoblastic leukemia / 中华实用儿科临床杂志

Objective To investigate Notch1 protein expression of leukemic cells in children with pediatric Bcell acute lymphoblastic leukemia(B-ALL) and its effect on prognosis.Methods The expression of Notch1 protein of leukemic cells in bone marrow smears was detected by immunohistochemistry(SABC) in primarily diagnosed childhood ALL,including 63 cases of B-ALL and 14 cases of T-ALL,a group of 34 children with no malignancy served as controls.Reverse transcription-ploymerase chain reaction was used to assay Notch1 mRNA expression in ALL patients.Results The incidence of Notch1 expression was 31.7％ in B-ALL patients,71.4％ in T-ALL patients,and 8.8％ in control group,respectively.Notch1 protein was aberrantly expressed in both B-ALL and T-ALL compared with the controls(P ＜ 0.05,P ＜ 0.001).Multivariate analysis revealed that the expression of Notch1 protein in B-ALL was not associated with patients' age,gender,WBC count at diagnosis(all P ＞ 0.05),it had no influence on the early treatment response.Nevertheless,the Kaplan-Meier curve of event-free survival showed that,in the patients without Notch1 protein expression,the long-term event-free survival rate was as high as 92.7 ％,in contrast,in the children with Notch 1 expression,the event-free survival was only 54.5 ％ (P =0.0054).Conclusions The expression of Notch 1 protein in pediatric B-ALL may predict a poor prognosis,and interfering with Notch1 signaling could be employed as a potential therapeutic target for those patients with Notch1 expression.

Biological characteristics of T-lineage acute lymphoblastic leukemia in 23 children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the biological characteristics of childhood T-lineage acute lymphoblastic leukemia (T-ALL) and their clinical significance.</p><p><b>METHODS</b>Immunophenotyping was performed by three-color flow cytometry analysis using CD45 /SSC gating in 23 children with newly diagnosed T-ALL. Meanwhile cytogenetic analysis was performed.</p><p><b>RESULTS</b>CD3(+) expression of T-lineage antigens was apparently higher than CD7(+) and CD5(+) expression. CD19(+) expression of B-lineage antigens was apparently higher than CD22(+), CD10(+) and CD20(+) expression. Myeloid antigen was expressed in 4 cases (17%). CD34(+) and HLA-DR(+) were observed in 4 cases (17%) and 5 cases (22%), respectively. cCD3(+) and cCD79(+) were expressed in 23 cases (100%) and 22 cases (96%), respectively. The chromosome detection in 8 cases with T-ALL showed hyperdiploid or Ph(+) chromosome (one case each). The fusion gene detection in 5 cases showed MLL rearrangements in two cases and positive SIL/TAL1 fusion gene in one case. CD3 expression was related with the complete remission rate.</p><p><b>CONCLUSIONS</b>Immunophenotyping is an important tool for diagnosis of T-ALL. However, the immunophenotype of T-ALL is heterogeneous. So, immunophenotyping along with cytogenetic and molecular genetic analysis is needed in the treatment and prognosis evaluation of T-ALL.</p>

Association of human parvovirus B19 infection and childhood idiopathic thrombocytopenic purpura: a meta analysis of Chinese literatures / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the relationship between human parvovirus B19 infection and childhood idiopathic thrombocytopenic purpura (ITP) by the principle of evidence based medicine.</p><p><b>METHODS</b>Papers related to the relationship between human parvovirus B19 infection and childhood ITP published between 1994 and 2008 were retrieved electronically from the Chinese Journals Full-text Database and the Wanfang Data. These relevant papers on case-control trials were statistically studied by meta analysis.</p><p><b>RESULTS</b>Eight papers that met the inclusion criteria were included for this meta analysis. Five hundred and sixteen cases of childhood ITP and 246 healthy controls were enrolled. The meta analysis showed that the incidence of human parvovirus B19 infection in the ITP group was significantly higher than that in the control group (OR=13.71, 95% CI=7.07-26.59, Z=7.75, p<0.01).</p><p><b>CONCLUSIONS</b>Human parvovirus B19 infection is closely associated with childhood ITP.</p>

Expression of CD147 and matrix metalloproteinase-9 in children with non-Hodgkin's lymphoma and its correlation with prognosis / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the expression of CD147 and matrix metalloproteinase-9 (MMP-9) in children with non-Hodgkin's lymphoma (NHL) and its correlation with clinical stage, tumor size, bone marrow invasion, immunological typing, serum lactate dehydrogenase (LDH) concentration, and prognosis.</p><p><b>METHODS</b>Specimens excised from NHL patients were prepared. Expression of CD147 and MMP-9 were tested by streptavidin-biotin complex (SABC) immunohistochemistry and its correlation with clinical results were analyzed.</p><p><b>RESULTS</b>The positive rate of CD147 expression was 73% (45/62), 17 cases were (-), 11 cases (+), 34 cases (++) and 21 cases (+++). The positive rate of MMP-9 expression was 81% (50/62), 12 cases were (-), 13 cases (+), 18 cases (++) and 19 cases (+++). The Spearman rank correlation analysis indicated that there was a positive correlation between CD147 and MMP-9 expressions in NHL (r(S) = 0.763, P = 0.034). Expression of CD147 was determined in relation to factors that included clinical bone marrow invasion, tumor size, LDH level as well as the clinical stage; expression of MMP-9 had a positive correlation with bone marrow invasion, tumor size and clinical phases. The 5-year survival rates (5YSR) were 78% (22/28) and 45% (15/34) in the cases whose CD147 expression was (-)-(+) and (++)-(+++), respectively, and 5YSR were 84% (21/25) and 43% (16/37) in the cases whose MMP-9 expression was (-)-(+) and (++)-(+++) respectively, the difference was significant. Cox multivariate analysis showed that both CD147 and MMP-9 were important prognostic factors.</p><p><b>CONCLUSION</b>The increased expression of CD147 and/or MMP-9 correlates with a poor clinical outcome in patients with NHL.</p>

Effects of SODD and survivin on leukemia cell apoptosis induced by chemotherapeutic drugs / 中国实验血液学杂志

This study was aimed to investigate the changes of silencer of death domains (SODD), survivin, caspase 3, caspase 8 and caspase 9 in the apoptotic process of human leukemia cells induced by chemotherapeutic drugs in order to explore the molecular mechanism of apoptotic modulatory genes and to search for the new target of chemotherapeutic drugs. After Jurkat cells were induced by chemotherapeutic drugs, the translocated phosphatidylserine was labeled with annexin V/PI, and the apoptosis incidence was measured by FCM; The expression changes of SODD, caspase 3, caspase 8 and caspase 9 were determined by Western blot; the changes of survivin mRNA and protein were determined by RT-PCR and immunohistochemistry SABC method respectively. The results indicated that high expressions of SODD and survivin could inhibit apoptotic signaling pathway; VCR down-regulated the function of SODD protein and effectively induced the apoptosis of Jurkat cells in a time-dependent manner and activates caspase 3 through the death receptor-mediated activation of caspase 8, in which caspase 9 and survivin were not degraded. It is concluded that SODD participates in the apoptotic process induced by VCR which induces the Jurkat cell apoptosis by downregulating expression of SODD protein and priming death receptor pathway. In the apoptotic process, the mitochondrion apoptotic pathway is not trigged.

Expression of survivin and its location in bone marrow cells of childhood acute leukemia: relationship to therapeutic efficacy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Survivin, a unique member of the inhibitor of apoptosis protein (IAP) family, plays an important role in regulating both apoptosis and cell division. Overexpression of survivin is associated with increased risk of recurrence and poor outcome in cancer patients. This study aimed to investigate the expression of survivin and its location as well as the relationship between cellular location and expression of survivin and the therapeutic efficacy at the cellular level.</p><p><b>METHODS</b>The expression of survivin protein was detected by immunohistochemical assay in bone marrow cells from 62 children with acute leukemia and 40 hospitalized children who did not have leukemia (Control group), and in a human acute T lymphocytic leukemia cell line (Molt-4 cells) treated in vitro with daunorubicin (DNR). Cell apoptosis was detected using flow cytometry.</p><p><b>RESULTS</b>Survivin protein was expressed in 41.9% of the 62 children with acute leukemia but in only 5.0% of the Control group (chi(2)=16.66; P < 0.01). The expression rate of survivin was 46.2% in cytoplasm and 53.9% in nucleus in the children with acute leukemia (chi(2)0.3077; P> 0.05). However, the remission rate of patients in whom survivin expression was seen in the nucleus was significantly higher than that in patients in whom survivin was expressed in cytoplasm after chemotherapy. The survivin expression in Molt-4 cells decreased remarkably by DNR treatment in a time and dosage-dependent manner. DNR treatment also induced survivin transllocation from cytoplasm to nucleus and cell apoptosis in a time and dosage-dependent manner.</p><p><b>CONCLUSIONS</b>Survivin may play an important role in the development and prognosis of childhood acute leukemia. The different expression pattern of survivin in the cytoplasm and the nucleus may be associated with therapeutic efficacy and prognosis in acute leukemia. DNR may reduce the survivin expression in leukemic cells and induce cell apoptosis.</p>

Regulation of NF-kappaB/P65 by MDM2 in acute lymphoblastic leukemia in childhood / 中华儿科杂志

<p><b>OBJECTIVE</b>MDM2 is considered a proto-oncogene due to its ability to inhibit P53 tumor-suppressor function. But, evidence showed that MDM2 might have a P53-independent role in tumorigenesis. MDM2 is over-expressed in human sarcoma and carcinoma. Recent studies showed that MDM2 might act as a transcriptional factor to modulate expressions of other genes involved in cell cycle regulation and transformation. In the present study, the investigators hypothesized that MDM2 directly affected NF-kappaB expression and function in a P53-independent manner.</p><p><b>METHODS</b>MDM2 was transfected to acute lymphoblastic leukemia (ALL) line EU-4 cells lacking P53 expression and expressing very low levels of MDM2. MDM2 and P65 expression in mRNA level and protein level were detected by Western blot and Northern blot after transfection. Since the expression of E-selectin is P65 dependent, E-selectin promoter-CAT construct and P65 and MDM2 expression plasmids were co-transfected to EU-4 cells. CAT activation was determined with ELISA. The effect of adriamycin (ADM) at the concentrations of 15 micro g/ml, 7.5 micro g/ml, 5 micro g/ml and 1 micro g/ml on MDM2-transfected EU-4 cells and the parent cells was detected by MTT assay.</p><p><b>RESULTS</b>The results showed that MDM2 up-regulated P65 expression at both mRNA and protein levels, and MDM2 increased P65-mediated transactivation of E-selectin promoter. Without P65, MDM2 had no effect on the transactivation of E-selectin. Moreover, MDM2 antisense could not change the transactivation of E-selectin. MTT results showed that the survival rate of MDM2 transfected EU-4 cells was higher than that of parental cells. The results suggested that MDM2 transfection increase drug resistance of EU-4 cells to ADM compared with parent cells.</p><p><b>CONCLUSION</b>(1) MDM2 up-regulated transcriptionally P65 expression. (2) MDM2 increased drug resistance of leukemia cells to ADM. (3) MDM2 elevated NF-kappaB activity in a P53-independent manner in childhood lymphoblastic leukemia cell line.</p>

Expressions of Silencer of Death Domains and p65 in Children with Acute Lymphoblastic Leukemia and Its Relationship with Chemotherapeutic Drugs / 实用儿科临床杂志

Objective To explore the expression of silencer of death domains(SODD) and its clinical significance and relationship with phospho-NF-?B-p65 proteins in bone marrow cells of acute lymphoblastic leukemia(ALL)in children,and the expression of SODD and phospho-NF-?B-p65 in Jurkat cells treated with chemotherapeutic drugs in order to find a new chemotherapeutic target.Methods The expressions of SODD and phospho-NF-?B-p65 proteins in bone marrow cells were detected by immunohistochemistry in 25 children with ALL.The apoptosis incidence was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-?B-p65 proteins were determined by Western blotting in Jurkat cells.Results It was found that the expression of SODD and active p65 expression in ALL were significantly higher than those in healthy control group.The expression of SODD and phospho-NF-?B-p65 proteins in the high-risk(HR) group was significantly higher than those in standard-risk(SR) group(Pa