[Effect of breastfeeding on immune function in infants with human cytomegalovirus infection]. / æ¯ä¹³å–‚å »å¯¹äººå·¨ç»†èƒžç— æ¯’æ„ŸæŸ“å©´å„¿å ç–«åŠŸèƒ½çš„å½±å“.

OBJECTIVES: To study the effect of breastfeeding on immune function in infants with human cytomegalovirus (HCMV) infection. METHODS: A retrospective analysis was performed on the medical data of 135 infants with HCMV infection who were admitted to Children's Hospital Affiliated to Zhengzhou University from January 2021 to May 2022, and all these infants received breastfeeding. According to the results of breast milk HCMV-DNA testing, the infants were divided into two groups: breast milk HCMV positive (n=78) and breast milk HCMV negative (n=57). According to the median breast milk HCMV-DNA load, the infants in the breast milk HCMV positive group were further divided into two subgroups: high viral load and low viral load (n=39 each). Related indicators were compared between the breast milk positive and negative HCMV groups and between the breast milk high viral load and low viral load subgroups, including the percentages of peripheral blood lymphocyte subsets (CD3+ T cells, CD3+CD4+ T cells, CD3+CD8+ T cells, and CD19+ B cells), CD4+/CD8+ ratio, IgG, IgM, IgA, and urine HCMV-DNA load. RESULTS: There were no significant differences in the percentages of CD3+ T cells, CD3+CD4+ T cells, CD3+CD8+ T cells, and CD19+ B cells, CD4+/CD8+ ratio, IgG, IgM, IgA, and urine HCMV-DNA load between the breast milk HCMV positive and HCMV negative groups, as well as between the breast milk high viral load and low viral load subgroups (P>0.05). CONCLUSIONS: Breastfeeding with HCMV does not affect the immune function of infants with HCMV infection.

[Effects of circ-SFMBT2 on Proliferation, Migration and Invasion of Acute Myeloid Leukemia Cells by Regulating miR-491-5p/HOXA9 Axis].

OBJECTIVE: To explore the effects and molecular mechanism of circ-SFMBT2 on the proliferation, migration and invasion of acute myeloid leukemia (AML) cells. METHODS: Bone marrow samples from 35 pediatric AML patients and 35 healthy controls in Henan Provincial Children's Hospital from April 2015 to April 2017 and human bone marrow stromal cell lines (HS-5) and AML cell lines (HL-60, THP-1, U-937 and Kasumi-1) were collected. The expressions of circ-SFMBT2, miR-491-5p and homeobox A9 (HOXA9) in bone marrow samples and cells were detected by RT-qPCR and Western blot. The Pearson method was used to analyze the correlation of circ-SFMBT2, miR-491-5p and HOXA9 mRNA expression levels in bone marrow samples of AML patients. HL-60 cells were cultured in vitro and divided into 5 groups: Control, si-NC, si-circ-SFMBT2, si-circ-SFMBT2+anti-NC and si-circ-SFMBT2+anti-miR-491-5p, HL-60 cells were transfected with si-NC, si-circ-SFMBT2, anti-NC, and miR-491-5p inhibitor with Lipofectamine™ 3000. RT-qPCR and Western blot were performed to detect the expression levels of circ-SFMBT2, miR-491-5p and HOXA9 in cells of each group. The proliferation activity of HL-60 cells in each group was detected by CCK-8 assay at 24, 48 and 72 h after transfection, respectively. The apoptosis rate was detected by flow cytometry. The migration and invasion abilities of cells were detected by Transwell assay. The regulatory roles of circ-SFMBT2, miR-491-5p and HOXA9 in AML cells were verified by dual-luciferase reporter gene assay, RNA pull-down and RNA-binding protein immunoprecipitation (RIP) experiments. RESULTS: The expression levels of circ-SFMBT2 and HOXA9 mRNA were increased in bone marrow samples and cell lines (HL-60, THP-1, U-937 and Kasumi-1) of children with AML (P <0.001), while the expression level of miR-491-5p was significantly decreased (P <0.001). Pearson correlation analysis showed that the expression levels of circ-SFMBT2 and miR-491-5p in bone marrow samples of AML children were negatively correlated (r =-0.905), miR-491-5p was also negatively correlated with HOXA9 mRNA (r =-0.930), while the expression levels of HOXA9 mRNA and circ-SFMBT2 was positively correlated (r =0.911). The overall survival rate of AML children with high expression of circ-SFMBT2 was significantly decreased than those with low expression of circ-SFMBT2 (P <0.05). Silencing of circ-SFMBT2 could greatly up-regulate the expression of miR-491-5p, decrease the expression of HOXA9, inhibit the proliferation, migration and invasion of AML cells, and promote cell apoptosis (P <0.05). Down-regulation of miR-491-5p expression greatly attenuated the inhibitory effects of circ-SFMBT2 silencing on cell proliferation, migration and invasion (P <0.05). Dual-luciferase reporter gene assay, RNA pull-down and RIP experiments confirmed that circ-SFMBT2 could target miR-491-5p and negatively regulate the expression of miR-491-5p in AML, and HOXA9 was the target of miR-491-5p. CONCLUSION: Silencing of circ-SFMBT2 may inhibit the proliferation, migration and invasion of AML cells by regulating the miR-491-5p/HOXA9 axis.

[Expression and Prognostic Impact of HK2 in Bone Marrow of Patients with Newly Diagnosed Acute Myeloid Leukemia].

OBJECTIVE: To detect the expression level of HK2 gene in the bone marrow of newly diagnosed patients with acute myeloid leukemia (AML) and investigate its influence on the clinical characteristics and prognosis. METHODS: The expression level of HK2 gene in the bone marrow of 90 newly diagnosed patients with AML that accompanying clinical characteristics and survival status were detected by RT-qPCR, and compared with 18 allogeneic hematopoietic stem cell transplantation (allo-HSCT) donors. The Chi-square test, Kaplan-Meier survival analysis, and Cox proportional hazards regression model were used to analyze the correlation of HK2 expression level with clinical characteristics and prognosis. RESULTS: Compared with allo-HSCT donors, the HK2 expression was significantly increased in newly diagnosed AML patients (P <0.01). Compared with patients with total response (OR, complete response + complete response with incomplete hematologic recovery) after 2 courses of induction chemotherapy, the expression of HK2 in patients without OR was significantly increased (P <0.05). There was a significant difference in the relative expression of HK2 between patients with and without OR after 2 courses of induction therapy (P <0.001). The median survival time of patients with high expression of HK2 was significantly shorter than that of patients with low expression of HK2 (P <0.05). The multivariate Cox proportional hazards regression analysis showed that prognostic stratification, the expression level of HK2, and whether two courses of induction therapy achieved OR were independent factors affecting the prognosis of AML patients (P <0.05). CONCLUSIONS: Compared with allo-HSCT donors, the expression level of HK2 gene is increased in the bone marrow of newly diagnosed AML patients. The prognosis of patients with high expression of HK2 is poor. The expression level of HK2 is an independent factor affecting the prognosis of AML patients.

[Effects of MiR-221-Mediated Wnt/ß- Catenin Signaling Pathway on Biological Activity of Childhood Acute Lymphoblastic Leukemia Cells].

OBJECTIVE: To study the effects of miR-221 on the biological activity of childhood acute lymphoblastic leukemia cells and its mechanism. METHODS: Bone marrow mononuclear cells (BMNC) were isolated from bone marrow samples of ALL children diagnosed in our hospital from May 2018 to November 2018. The cells were divided into control group, miR-221-NC group and miR-221 group. After transfection according to the instructions of Lipofectamine 2000 kit, the levels of miR-221 in each group were detected by RT-PCR. Flow cytometry was used to detect the effects of miR-221 on cell cycle and apoptosis. Transwell assay was used to detect cell migration and invasion. Western blot was used to detect the effects of miR-221 on proliferating cell nuclear antigen (PCNA), Caspase 3, Cyclin D1 and MMP-9 proteins in BMNC. Luciferase reporter gene assay was used to detect the targeting relationship between miR-221 and Wnt gene. RESULT: The expression level of miR-221 in the miR-221 group was significantly higher than that in the control group and the miR-221-NC group (P＜0.05). MTT assay showed that, after transfection for 2, 3, 4 and 5 days, the cell proliferation level in miR-221 group was significantly lower than that in the control group and the miR-221-NC group (P＜0.05). The cell ratio of G0/G1 phase was (73.25±8.1)% in the miR-221 group, which was significantly higher than that in the control group and the miR-221-NC group (P＜0.05); moreover, the cell ratio of S phase in the miR-221 group was (12.37±1.6)%，which was significantly lower than that in the control group and the miR-221-NC group (P＜0.05). The percentage of apoptotic cells in the miR-221 group was (24.68±3.87)%, which was significantly higher than that in the control group and the miR-221-NC group (P＜0.05). Transwell cell invasion experiment showed that the number of invasive cells in the miR-221 group was 23.42±3.62, which was significantly lower than that in the control group and the miR-221-NC group (P＜0.05). Transwell cell migration assay showed that the number of migrating cells in the miR-221 group was 34.86±5.32, which was significantly lower than that in the control group and the miR-221-NC group (P＜0.05). The relative level of PCNA, Cyclin D1 and MMP-9 in the miR-221 group was 0.26±0.03, 0.17±3.61 and 0.14±0.02, respectively, which was significantly lower than those in the control group and the miR-221-NC group (P＜0.05), while the relative level of Caspase-3 in the miR-221 group was 0.37±0.05, which was significantly higher than that in the control group and the miR-221-NC group (P＜0.05). Luciferase reporter assay showed that the activity of luciferase in Wnt wild type plasmid was significantly inhibited by miR-221 (P＜0.05). CONCLUSION: miR-221 can inhibit the proliferation, migration and invasion of BMNC, moreover can promote cell apoptosis, which may be related with the inhibition of Wnt/ß- catenin signaling pathway.

[Clinicopathological Analysis of Children's Systemic EBV-Positive T-Cell Lymphoma].

OBJECTIVE: To explore the clinical and pathologic features as well as prognosis of systemic EBV-positive T-cell lymphoma in children. METHODS: The clinical data including clinical manifestation, pathologic changes and treatment in 16 patients with children's systemic EBV-positive T-cell lymphoma were analyzed retrospectively, and follow-up of patients were carried out. RESULTS: The 16 cases included 12 males and 4  females with median age of 3.3 years old. It was demonstrated that the clinical and pathological features of the children's systemic EBV-positive T-cell lymphoma were as followed fever, hepatosplenomegaly, cytopenia, lymphadenopathy, and hemophagocytosis in bone marrow or organ. Histologically, the structures of lymph node was normal, partially or completely destoryed. The paracortical zone was expanded with prominent infiltration of small to medium-sized atypical lymphocytes. The major immunophenotypic characteristics were as follows: (1) Almost all biopsies exhibited prominent T cell proliferation. (2) CD3 was expressed in 16 patients (100%, 16/16), CD4 in 5 patients (31.3%, 5/16)，CD5 in 13 patients (81.3%, 13/16)，CD7 was expressed in 11 patients (68.8%, 11/16)，CD8 in 15 patients (93.8%, 15/16)，CD4 and CD8 were expressed in 5 patients (31.3%, 5/16)，CD4 and CD8 double-negative in patients (6.3%, 1/16)，16 patients were CD56 negative (100%, 16/16). (3) TCR gene cloning rearrangement in 16 patients (93.8%, 15/16). (4) EBV-EBER was expressed in 16 patients (100%, 16/16). 11 out of 16 cases died, 1 cese failed to be followed up, 1 case relapsed，and 3 cases survived, reseptively. The media survival time was 4 months. CONCLUSION: Systemic EBV-positive T-cell lymphoma predominantly occurred in childhood and early teen-age, and lacks specific clinic features, usually combined with hemophagocytic syndrome. The confirmed diagnosis requires comprehensive analysis of clinical manifestation, pathomorphology, immunohistochemical detection, EBV-EBER insite hybridization, and TCR gene test. The overall prognosis of the disease is poor and the fatality rate is high.

Withaferin A activates stress signalling proteins in high risk acute lymphoblastic leukemia.

Withaferin A, the principal bio-active component isolated from the Withaniasomnifera, has shown promising anti-leukemic activity in addition to anti-invasive and anti-metastatic activity. The present study demonstrates the effect of withaferin A on the cell cycle status and the phosphorylation/activation of proteins involved in signal transduction in t(4;11) and non-t(4;11) acute lymphoblastic leukemia (ALL) cell lines after treatment with withaferin A. The cells after treatment with the vehicle or 25 µM withaferin A for 1, 2, 4 and 8 h were examined using flow cytometric analysis. The results revealed that withaferin A treatment induced cell growth arrest at the S to G2/M phase transition of the cell cycle. Withaferin A treatment also induced the phosphorylation of stress signalling proteins, including the p38 mitogen-activated protein kinase, the c-Jun N-terminal kinase, c-Jun, the heat shock protein 27 and protein kinase B within 0 to 16 h. These results were observed using multiplex technology and Western blotting analysis. Thus withaferin A induces stress response leading to cell death. Therefore, withaferin A can be a potent therapeutic agent for the treatment of high risk ALL with chromosomal translocation t(4;11).

Synthesis and biological activity of novel tiliroside derivants.

A series of new tiliroside derivatives were synthesized and characterized by analytical (1)H NMR, (13)C NMR and mass spectrometry. All of the compounds were evaluated for anti-diabetic properties in vitro using HepG2 cells. Compounds 3c, 3d, and 3i-l caused significant enhancements in glucose consumption by insulin-resistant HepG2 cells compared with control cells and cells that were exposed to metformin (an anti-diabetic drug). Moreover, compound 3l significantly activated adenosine 5'-monophosphate-activated protein kinase activity and reduced acetyl-CoA carboxylase activity. Thus, the tiliroside derivative 3l offers potential to be developed as a new approach for treating type II diabetes.