Expression and Significance of Vitamin D Receptor Gene and NF-κB Pathway in Blood of Children with Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression, clinical significance and prognosis of vitamin D receptor (VDR) gene and NF-κB pathway in children with acute lymphoblastic leukemia (ALL).@*METHODS@#Thirty children with definitive diagnoses of ALL from December 2018 to December 2021 were selected as ALL group, and 30 healthy children under physical examination were selected as control group. Peripheral blood of all study subjects was collected. The VDR and NF-κB mRNA and protein expressions were detected by real-time quantitative PCR and Western blot, respectively. The relationship between mRNA expression of the above genes and clinical characteristics of children was retrospectively analyzed.@*RESULTS@#The relative expression of VDR mRNA in peripheral blood of children with ALL was significantly lower than that in the control group (P < 0.05), while NF-κB mRNA was higher (P < 0.001). The expression of NF-κB mRNA in ALL children with peripheral blood white blood cell count (WBC) < 50×109/L at initial diagnosis was significantly higher than those with WBC≥50×109/L (P < 0.01). The expression of NF-κB mRNA in ALL children with infection was significantly higher than that those without infection (P < 0.05). There were no significant differences in NF-κB mRNA expression between children with different sex, age, hemoglobin at initial diagnosis, platelet, immunologic typing, risk and induced response (P >0.05).@*CONCLUSION@#The expression of NF-κB is of value to diagnosis and prognosis of ALL in children.

Genomic and Clinical Analysis of Children with Acute Lymphoblastic Leukemia.

Objective: This study investigated the types and significance of mutant genes in children with acute lymphoblastic leukemia (ALL). Methods: The gene sequencing data of 89 ALL children were retrospectively analyzed. Log-rank test was used to analyze the effect of different numbers of mutant genes on the clinical characteristics of the patients and disease. Results: Known gene mutations were detected in 64% (57/89) of the children, including one gene mutation in 31% and two or more gene mutations in 33% of the patients. Gene sequencing showed that most mutations occurred in KRAS (17%), NRAS (15%), FLT3 (7%), TP53 (7%), and PTPN11 (7%), and functional clustering analysis showed that most were signaling pathway genes (50%). In the overall cohort, no association was found between clinical characteristics and gene mutation. The children were then classified into three groups: group A (no gene mutation), group B (one gene mutation), and group C (two or more gene mutations). Correlation analysis showed that group A had significantly more children with medium risk ALL (P = 0.037), and group C had markedly more children with high risk ALL (P = 0.001). Further analysis showed that children with mutant genes took significantly more time to enter the maintenance phase than children without mutations. Conclusion: Children with ALL had a high gene mutation rate, especially in KRAS and NRAS genes, and the mutant genes were mainly signal pathway-related. The gene mutations were significantly correlated with clinical phenotype and the time taken to enter the maintenance phase.

Fos-Related Antigen 1 May Cause Wnt-Fzd Signaling Pathway-Related Nephroblastoma in Children.

This study aimed to investigate the role of the primary Fos-related antigen 1 (Fosl-1) oncogene in nephroblastoma by studying 60 childhood nephroblastoma and 58 paraneoplastic carcinoma cases. The Fosl-1 expression was detected using immunohistochemistry. In vitro culture of nephroblastoma cells was performed by viral transfection to establish Fosl-1 overexpression and gene knockout models. Flow cytometry and nano-PCR were used to detect apoptosis and mRNA expression in related pathway genes. Immunohistochemical results showed that the positive expression of Fosl-1 in the nuclei of nephroblastoma tissue was 78%, among which metastasis rate was 61.7%; correspondingly, it was 8%, and 100% in adjacent tissues. The qPCR results indicated that MMP9, Wnt1, and Fzd1 were significantly upregulated after Fosl-1 overexpression compared with the normal embryonic tissue cells, control, and gene knockout groups (P <0.05). Fosl-1 could cause the occurrence, development, and metastasis of childhood nephroblastoma through wingless/int1/Frizzled-related signaling pathways.

The reversal effect of abnormal savda munziq total flavonoids on leukemia cell lines / 中国基层医药

Objective To investigate the reversal effect of abnormal savda munziq total flavonoids on leukemic cell lines.Methods CCK -8 method was used to detect the cytotoxicity of abnormal savda munziq total flavonoids on leukemia cells,by multivariate analysis of variance in the chemotherapy drug intervention,further calculated the reversal fold.Results CCK -8 results showed that with the increase of concentration of ASMq800(μg/mL)compared with 200(μg/mL)and 400(μg/mL),the A value of K562 /ADR cells gradually decreased,the growth curve suggested that with the increase of ASMq concentration in K562 /ADR cell,survival time was shortened,the inhibition of the cells was significantly increased.Different concentrations of ASMq on the adriamycin toxicity with increase of concen-tration of adriamycin,the inhibition of ASMq on multidrug resistance cell line was significantly higher.The reversal multiples calculated that with the increase of ASMq concentration 600 μg/mL and 900 μg/mL,the reversal index increased significantly 2.485 times and 3.651 times.Conclusion ASMq on human leukemia multidrug resistance cell line K562 /ADR could reverse the drug resistance,the reversal mechanism needs further study.

Reversal effect study of vitamin D on leukemia multidrug resistance cells / 中华实用儿科临床杂志

Objective To investigate the reversal effect of vitamin D on drug resistance leukemia cell lines Jurcat vitamin D/ADR,K562/ADR and its mechanism.Methods Non-toxic concentration of vitamin D was chosen by methyl thiazolyl tetrazolium,and on line concentration experiment ;the contents of vitamin D for soft erythromycin in the cell and the cell surface of P-glycoprotein(P-gp) expression level were detected by using flow cytometry instrument and vitamin D for intracellular mdr1 mRNA and mrp1 mRNA expression was detected by Real-time polymerase chain reaction;vitamin D for the change of intra cellular glutathione glycosides peptide (GSH) content was detected by adoping biochemistry method.Results Resistance reversal in experiments showed that vitamin D could decrease soft erythromycin in the accumulation of drug resistant cells,with a dose-dependent relationship; vitamin D treatment Jurcat/ADR,K562/ADR cells after 48 h,chemotherapy drugs could increase the cell concentration,and reduce P-gp and the expression of mdr1 and mrp1 gene ; vitamin D treatment Jurcat/ADR,K562/ADR after 48 h could decrease the intracellular GSH content in K562,K562/ADR,Jurcat and Jurcat/ADR.Conclusions Vitamin D has the drug resistance reversal agents to Jurcat/ADR,K562/ADR,which might be relevant to the inhibition of cell surface of P-gp proteins function and expression and reduction of the mdr1 and mrp1 expressions of resistance genes and reduction of intracellular GSH content.Vitamin D by influencing the above mechanism,which increases the drug concentration in cells,can effectively kill the tumor cells.

[Effects of Huangqi injection on infection factors in children with acute lymphoblastic leukemia].

OBJECTIVE: To investigate the effects of Huangqi injection on the infection factors in children with acute lymphoblastic leukemia (ALL) during remission induction chemotherapy. METHODS: Ninety-one children with ALL were divided into treatment (n=47) and control groups (n=44) by a randomized double-blind method. During remission induction chemotherapy, the treatment group was given Huangqi injection (0.5 mL/kg·d) for 35 days, while an equal volume of normal saline was used instead in the control group; the other supportive care was the same for the two groups. After remission induction chemotherapy, the incidence of infection, duration of infection, white blood cell and neutrophil counts, site of infection, and positive rate of pathogenic bacteria in secretion were compared between the two groups. RESULTS: Four cases in the treatment group dropped out of the study due to allergic reaction. After remission induction chemotherapy, compared with the control group, the treatment group had a significantly lower incidence of infection (P<0.05), a shorter duration of infection at any site (P<0.05), a higher neutrophil count after chemotherapy (P<0.05), and lower incidence rates of respiratory tract infection, urinary tract infection, blood infection, and skin and soft tissue infections (P<0.05). Gram-negative bacteria were the main pathogens. Among the infected children, the positive rate of pathogenic bacteria in secretion was significantly lower in the treatment group than in the control group (P<0.05). CONCLUSIONS: Huangqi injection may reduce bone marrow suppression caused by chemotherapy drugs and increase neutrophil count during remission induction chemotherapy to reduce the incidence and duration of infection in children with ALL.

Immunophenotyping of leukemic stem cells and chromosome karyotype characteristics in Uyghur leukemia pediatric patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the immunophenotype and chromosome karyotype characteristics of leukemic stem cells (LSC) in Uyghur leukemia pediatric patients.</p><p><b>METHODS</b>The morphological features of LSC in culture in vitro was observed by flow cytometry. The immunophenotype was assessed by detective flow cytometry. The chromosome karyotype was analyzed by R-banding technique.</p><p><b>RESULTS</b>The LSC showed suspended floating colonies growing in the culture medium, and grew well and proliferated constantly in culture over 8 months. Among the 13 children with AML, there were 10 CD34(+)CD38(-)CD123(+) and CD33(+) cases, 10 CD44(+) cases, 10 CD96(+) cases, and 5 CD90(+) cases. Among the 13 children with B-ALL, there were 6 CD34(+)CD20(-)CD19(+) cases, 7 CD9(+) cases, and 5 CD123(+) cases. Among the 9 children with acute T lymphoblastic leukemia (T-ALL), there were 5 CD34(+)CD7(-) and CD90(+) cases, and 4 CD123(+) cases. Among the 13 cases of AML, 5 cases showed chromosome translocation t(15;17), one case chromosome translocation t(8;21), and 7 cases showed no chromosome karyotype abnormality. Among the 22 ALL cases, there were chromosome translocation t(12;21) in 1 case, t(9;22) in 3 case, hyperdiploid in 2 cases, and 16 cases without karyotype abnormalities. Twenty-nine children received induction remission therapy. Among them, 12 died, including 9 CD96(-)positive cases and 3 CD96(-)negative cases, with a statistically significant difference (P < 0.05).</p><p><b>CONCLUSIONS</b>The LSC of Uyghur leukemia pediatric patients in Xinjiang express CD9 and CD19 in ALL, and express CD123 and CD90 simultaneously in ALL and AML. The expression of CD96 is one of factors of poor prognosis.</p>

[Adverse effects of methotrexate in the treatment of acute lymphoblastic leukemia in Uyghur and Han children].

OBJECTIVE: To study blood concentrations of methotrexate (MTX) in Uyghur and Han children with acute lymphoblastic leukemia (ALL), and to provide criteria for judging the incidence of adverse effects of MTX. METHODS: Twenty-eight children with ALL (15 Han children and 13 Uyghur children), who received high-dose MTX chemotherapy, were divided into >10 µmol/L and ≤10 µmol/L groups according to 24-hour blood concentration of MTX, and divided into >1.0 µmol/L and ≤1.0 µmol/L groups according to 48-hour blood concentration of MTX. Enzyme multiplied immunoassay was used to measure blood concentrations of MTX in the MTX-treated children at 24 and 48 hours after MTX administration, and the adverse effects were observed. RESULTS: There was no significant difference in the incidence of adverse effects between the >10 µmol/L and ≤10 µmol/L groups (P>0.05). The >1.0 µmol/L group showed higher incidences of gastrointestinal reactions and mucosal injuries than the ≤1.0 µmol/L group (P<0.05), but no significant difference was found between the two groups with respect to the incidence of abnormal liver function and bone marrow suppression (P>0.05). Compared with Uyghur children, Han children showed higher 24- and 48-hour blood concentrations of MTX (P<0.05) and higher incidence of abnormal liver function, mucosal injuries, and bone marrow suppression (P<0.05). CONCLUSIONS: The 24-hour blood concentration of MTX cannot be used to predict the incidence of adverse effects in MTX chemotherapy, but 48-hour blood concentration of MTX is helpful in this regard. There are significant differences in 24- and 48-hour blood concentrations of MTX and the incidence of adverse effects between Uyghur and the Han children with ALL who receive MTX chemotherapy. Monitoring of blood MTX concentration maybe significant for timely adjustment of MTX dosage and individualized MTX chemotherapy.