[Effect of polydatin on the proliferation and apoptosis of THP-1 cells and the mechanism]. / 虎杖苷对THP-1ç»†èƒžå¢žæ®–åŠå‡‹äº¡çš„å½±å“åŠå ¶ä½œç”¨æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To explore the effect of polydatin on the proliferation and apoptosis of acute monocytic leukemia cell line THP-1 and the possible mechanism. METHODS: After THP-1 cells were treated with polydatin at gradient concentrations for 24 hours and 48 hours, their proliferation was determined by CCK-8 assay, and half maximal inhibitory concentration (IC50) was calculated. Logarithmically growing THP-1 cells were divided into two groups, a polydatin treatment group (treated with IC50 of polydatin) and a blank control group (treated without polydatin solution), and incubated for 48 hours. Cell apoptosis and cell cycle were measured by flow cytometry. The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins were measured by Western blotting. RESULTS: After treatment with polydatin, the proliferation of THP-1 cells was strongly inhibited, and the IC50 at 48 hours was 1 800 µmol/L. After treatment with 1 800 µmol/L polydatin solution for 48 hours, the apoptosis rate of THP-1 cells increased significantly compared with the blank control group (P<0.05). The cell cycle was arrested in the G0/G1 and S phases, with a significantly increased proportion of cells in the G0/G1 phase and a significantly decreased proportion of cells in the S phase, as compared with the blank control group (P<0.05). The expression levels of PI3K, AKT, p-AKT, mTOR, p-mTOR, p70 S6K, and p-p70 S6K proteins decreased significantly compared with the blank control group (P<0.05). CONCLUSIONS: Polydatin can effectively inhibit the proliferation, block the cell cycle, and induce the apoptosis of THP-1 cells, which may be related to inhibition of the PI3K/AKT/mTOR signaling pathway.

MAGI2-AS3 restrains proliferation, glycolysis, and triggers apoptosis in acute lymphoblastic leukemia via regulating miR-452-5p/FOXN3 pathway.

Objectives: MAGI2-AS3 is a cancer suppressor gene of multiple malignancies. Acute lymphoblastic leukemia (ALL) is an important type of leukemia that especially occurs in children. Our work evaluated the modulation of MAGI2-AS3 in ALL. Materials and Methods: qPCR and Western blotting were adopted for detection of target molecular expression. Growth and apoptosis were determined by CCK8 assay and Annexin V/PI staining. Glycolysis was detected by commercial kits. The direct binding between miR-452-5p and MAGI2-AS3 or FOXN3 was assessed by luciferase reporter assay. Tumor growth was measured in nude mice in vivo. Results: MAGI2-AS3 was down-regulated in ALL. Enforced expression of MAGI2-AS3 inhibited growth and glycolysis while promoting apoptosis of ALL cells. Moreover, MAGI2-AS3 up-regulated FOXN3 via sponging miR-452-5p. FOXN3 depletion abrogated MAGI2-AS3-mediated anti-cancer action. More importantly, MAGI2-AS3 repressed ALL cell growth in nude mice through regulation of miR-452-5p/FOXN3. Conclusion: MAGI2-AS3 inhibits ALL development via modulating miR-452-5p/FOXN3.

[Clinical features and prognosis of malignancy-associated hemophagocytic lymphohistiocytosis in children: a clinical analysis of 24 cases].

OBJECTIVE: To investigate the clinical features and prognosis of malignancy-associated hemophagocytic lymphohistiocytosis (MAHS) in children. METHODS: A retrospective analysis was performed for the primary diseases, clinical features, and prognosis of 24 children with MAHS. RESULTS: Among the 24 children, 11 (46%) had MAHS induced by tumor and 13 (54%) had chemotherapy-associated MAHS. As for primary diseases, 17 children had acute leukemia, 6 had lymphoma, and 1 had neuroblastoma. The most common clinical manifestations were pyrexia, respiratory symptoms, and hepatosplenomegaly. The most common laboratory abnormalities were hemocytopenia, elevated serum ferritin, and elevated lactate dehydrogenase. Of the 24 children, 22 were treated according to the HLH-2004 protocol and 2 gave up treatment; 18 children died, 1 was lost to follow-up, and 5 survived. The survival time ranged from 3 days to 2 years and 4 months (median 28 days). CONCLUSIONS: Children with MAHS have various clinical features and extremely poor treatment outcomes.

[Roles of follicular helper T cells and follicular regulatory T cells in pathogenesis of Henoch-Schönlein purpura in children].

OBJECTIVE: To study the roles of follicular helper T (Tfh) cells and follicular regulatory T (Tfr) cells in the pathogenesis of Henoch-Schönlein purpura (HSP) in children. METHODS: Peripheral blood samples were collected from 40 HSP children and 25 healthy controls. The percentages of Tfh and Tfr cells were measured by flow cytometry; the mRNA expression levels of Bcl-6, c-MAF, Blimp-1, and PD-1 in peripheral blood were measured by real-time polymerase chain reaction. RESULTS: Compared with the controls, the children with HSP had significantly increased percentage of Tfh cells and Tfh/Tfr ratio but a significantly reduced percentage of Tfr cells in the peripheral blood (P<0.05). Compared with the controls, the children with HSP had significantly increased mRNA expression of Bcl-6 and c-MAF but significantly reduced mRNA expression of Blimp-1 in CD4+ T cells (P<0.05), and had significantly increased mRNA expression of PD-1 but significantly reduced mRNA expression of Blimp-1 in CD4+CD25+ regulatory T cells (P<0.05). CONCLUSIONS: Abnormal percentages of Tfh and Tfr cells may be involved in the pathogenesis of HSP in children, and over-expression of Bcl-6, c-MAF, and PD-1 mRNA and inhibited expression of Blimp-1 mRNA may be considered as important reasons for abnormal percentages of Tfh and Tfr cells.

[Expression of CD58 in childhood B-lineage acute lymphoblastic leukemia and its feasibility in minimal residual disease detection].

OBJECTIVE: To measure the expression of lymphocyte function-associated antigen-3 (CD58) in childhood B-lineage acute lymphoblastic leukemia (B-ALL) and to explore the feasibility of CD58 as an indicator for minimal residual disease (MRD) detection in childhood B-ALL. METHODS: Eighty-seven children diagnosed with B-ALL between January 2014 and September 2014 were enrolled, and 20 hospitalized children who had no tumor or blood disease and had normal bone marrow cell morphology served as the control group. The expression features of CD58 in bone marrow samples from the two groups (at diagnosis, on day 15 of induction chemotherapy) were analyzed by four-color flow cytometry (FCM). Quantitative real-time polymerase chain reaction (qRT-PCR) and FCM were used to detect MRD in B-ALL patients on day 33 of induction chemotherapy. RESULTS: The mean fluorescence intensity of CD58 expression in the 87 B-ALL cases (91±33) was significantly higher than that in the 20 controls (14±6) (P<0.01); CD58 was over-expressed in 44 of the B-ALL cases. In the B-ALL children, the expression of CD58 on day 15 of induction chemotherapy (105±22) was not significantly different from that at diagnosis (107±26) (P>0.05). In the 44 B-ALL patients with CD58 over-expression, FCM showed 9 MRD(+) cases and 35 MRD(-) cases, while qRT-PCR showed 11 MRD(+) cases and 33 MRD(-) cases; 42 cases (95%) showed consistent results of the two tests, so there was no significant difference between the two methods in detecting MRD (P>0.05). CONCLUSIONS: CD58 is over-expressed and stable in children with B-ALL, and it can be considered as an indicator for MRD detection in childhood B-ALL.

[Clinical features of childhood hemophagocytic syndrome and its association with human parvovirus B19 infection].

OBJECTIVE: To investigate the association of childhood hemophagocytic syndrome (HPS) with human parvovirus B19 (HPVB19) infection, and to analyze the clinical features of this disease. METHODS: ELISA and quantitative real-time PCR were used to detect HPVB19-IgM, HPVB19-IgG and HPVB19-DNA in 65 children with HPS (HPS group) and 65 healthy children (control group). The HPS group was divided into HPVB19-infected (n=14) and non-infected (n=51) groups according to the detection results of HPVB19-DNA. The clinical data of two groups were compared. RESULTS: The positive rate of HPVB19-IgM in the HPS group (26%, 17/65) was significantly higher than that in the control group (9%, 6/65) (P=0.011), and there was no significant difference in the positive rate of HPVB19-IgG between the HPS (38%, 25/65) and control groups (29%, 19/65) (P=0.266). The infection rate of HPVB19 in the HPS group (22%, 14/65) was significantly higher than that in the control group (3%, 2/65) (P=0.001). Compared with the non-infected group, the HPVB19-infected group had significantly lower platelet count and hemoglobin level on admission, significantly more severe liver function damage, a significantly earlier onset time, and a significantly longer course of disease (P<0.05). CONCLUSIONS: The pathogenesis of HPS may be associated with HPVBl9 infection. HPVBl9-infected children with HPS have more acute onset, more severe clinical manifestations, and a longer disease duration.

[Prognostic significance of coagulation disorders in children with hemophagocytic syndrome].

OBJECTIVE: To study the prognostic significance of coagulation disorders in children with hemophagocytic syndrome (HPS). METHODS: Thirty-five children with HPS were retrospectively studied to analyze the etiology, clinical characteristics, laboratory results and treatment outcomes. RESULTS: After treatment, 27 of the 35 HPS patients survived, and the other 8 cases died. All cases were treated according to the HLH-2004 protocol, but etoposide (VP-16) was not used in 10 of them. The response rate in patients who received VP-16 (22/25, 88%) was significantly higher than that in those not receiving VP-16 (5/10, 50%) (P<0.05). Compared with the survival group, the dead group had significantly lower platelet count, fibrinogen level, and VP-16 utilization rate (P<0.05) but significantly longer activated partial thromboplastin time and prothrombin time (P<0.05). CONCLUSIONS: Coagulation function can be used as an indicator of disease outcome. It is essential for improving the clinical outcome of HPS to monitor the coagulation function during treatment, detect and correct abnormalities in time, and provide treatment strictly according to the HLH-2004 protocol.

Application of NT-proBNP in ventilator weaning for preterm infants with RDS.

OBJECTIVE: To evaluate the value of N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels along with spontaneous breathing trial (SBT) in the prediction of ventilator weaning outcome among respiratory distress syndrome (RDS) preterm infants ready to wean. METHODS: NT-proBNP along with plasma albumin concentration, serum sodium, serum potassium, and hematocrit were measured immediately before SBT in preterm infants (≤32 weeks) mechanically ventilated due to RDS. Extubation was considered successful if infants remained extubated >48 hr. Either SBT failure or extubation failure was considered weaning failure. RESULTS: Sixty-three of 88 infants passed the SBT and were subsequently extubated. Of these, two (3.2%) cases rapidly developed laryngeal dyspnea imposing reintubation (excluded from analysis). Of the remaining 61 infants, 45 (73.8%) cases had successful extubation, and 16 (26.2%) cases were reintubated. Infants who failed weaning had lower gestational age, birth weight, and plasma albumin concentrations, higher NT-proBNP, doses of surfactant, occurrence of ventilator-associated pneumonia, and occurrence of pulmonary arterial hypertension than those who did not. NT-proBNP was the only independent factor that could predict weaning failure (OR = 1.872; P = 0.044). The ROC-AUC for NT-proBNP to predict weaning failure was 0.977 (95% CI 0.918-0.997; P < 0.001). The cut-off of NT-proBNP level 18,500 pg/ml to predict weaning failure had a positive likelihood ratio of 25.180. The addition of NT-proBNP to SBT in prediction of weaning failure significantly improved the net reclassification improvement (NRI = 0.224; P = 0.034). CONCLUSION: NT-proBNP is an independent factor that could predict weaning failure. Measurement of NT-proBNP prior to SBT may be helpful in promoting successful ventilator weaning along with SBT.

[Latest update on immunotherapy of Epstein-Barr virus-associated post-transplantation lymphoproliferative disease].

Epstein-Barr virus (EBV)-associated post-transplantation lymphoproliferative disease (EBV-PTLD) is a potentially life-threatening complication after hematopoietic stem cell transplantation or solid organ transplantation. In the last decade, the survival of patients with EBV-PTLD has been significantly improved by immunotherapeutic interventions among high-risk patients. The immunotherapeutic interventions for EBV-PTLD include reduction in immunosuppression, CD20 monoclonal antibodies (rituximab) as monotherapy or in combination with chemotherapy, and adoptive immunotherapy with EBV-specific T cells. This paper reviews the latest update on the high-risk factors, clinical manifestations and immunotherapy of EBV-PTLD.

[Effectiveness of anti-CD47 antibody and Ara-C combination targeting therapy NOD/SCID mouse of myeloid leukemia].

OBJECTIVE: To study the prognostic significance of CD47 in a NOD/SCID mouse model of acute myeloid leukemia (AML) and the best strategy for targeted therapy for this disease. METHODS: CD34(+)CD38(-) leukemia stem cells (LSCs) were separated and transplanted into NOD/SCID mice to establish a mouse model of acute monocytic leukemia (AMoL). Anti-human CD47 antibody, alone or combined with cytosine arabinoside (Ara-C), was used to treat the mice with AMoL for 7-14 days, and therapeutic efficacy was assessed. LSCs were cultured together with mouse macrophages in culture medium containing anti-CD47 or anti-CD45 monoclonal antibody for 2 hours, to observe the phagocytic ability of macrophages to LSCs. RESULTS: CD34(+)CD38(-) LSCs existed among THP-1 cells, with a content of about (0.12 ± 0.06)%, and a mouse model of AML was successfully established after the purified CD34(+)CD38(-) LSCs (97.0% ± 1.7%) were transplanted into NOD/SCID mice. The in vivo experiment showed that mice with AMoL had the most significant decrease in CD33(+)CD45(+) leukemia cells in peripheral blood and bone marrow and survived the longest after being treated with Ara-C (7 days) plus anti-CD47 monoclonal antibody (14 days) (P < 0.01). After 2 hours of in vitro culture, the phagocytic index in the culture medium containing anti-CD47 monoclonal antibody was significantly higher than in the culture medium containing anti-CD45 monoclonal antibody (76.9% ± 12.2% vs 7.60% ± 2.4%; P < 0.05). CONCLUSIONS: High expression of CD47 is an adverse prognostic factor in AML. Combination therapy with anti-CD47 monoclonal antibody and Ara-C can effectively eliminate leukemia cells and LSCs, demonstrating great clinical significance in curing AML.

Association between helicobacter pylori infection and chronic idiopathic neutropenia.

The possible association between Helicobacter pylori (H. pylori) infection and chronic idiopathic neutropenia (CIN) was investigated. A total of 78 subjects with CIN were recruited in this case-control study. As a control group, 40 subjects without CIN were selected for comparison with the case group. All participants were evaluated for the prevalence of H. pylori infection by 14C-urea breath test. The corrected splenic index (CSI) was calculated, and serum IL-6, IL-8, IL-10 and HsCRP levels were measured. The differences in CSI, serum IL-6, IL-8, IL-10 and HsCRP levels were compared between CIN patients and controls, as well as between subjects with and without H. pylori infection. The positive rate of H. pylori was 87.18% in CIN group and 52.50% in control group, showing a significant difference (Fisher's exact, P=0.000). CSI values, and serum IL-6 and HsCRP levels in H. pylori positive-CIN patients were significantly higher than those in negative subjects (Mann-whitney U-test, P=0.016, P=0.001 and P=0.000 respectively), while IL-10 level declined significantly in H. pylori negative-CIN patients (Mann-whitney U-test, P=0.000). In control group, serum IL-6 and HsCRP levels in H. pylori positive individuals were also increased significantly (Mann-whitney U-test, P=0.000), while IL-10 level declined (Mann-whitney U-test, P=0.018). Multivariate regression analysis revealed that H. pylori infection and IL-10 were significant risk factors for CIN with odds ratio (OR): 3.09, 95.0% CI: 1.22-6.93; P=0.019, and OR: 0.17, 95.0% CI: 0.05-0.94; P=0.021, respectively. This prospective study confirmed the existence of an association between H. pylori infection and CIN, suggesting the screening for H. pylori infection and eradicating bacterium in positive cases seem appropriate and beneficial for those patients with CIN diagnosis.

[Role of Helicobacter pylori infection in the pathogenesis and clinical outcome of childhood acute idiopathic thrombocytopenic purpura].

OBJECTIVE: To investigate the role of Helicobacter pylori (Hp) and its products cytotoxin-associated protein (Cag A), vacuolating cytotoxin (VacA) in childhood acute idiopathic thrombocytopenic purpura (aITP), to evaluate the effect of Hp on their clinical outcome. METHODS: Subjects were enrolled according to case-control design, including 184 aITP children and 154 healthy controls. They were inquired for demographic characteristics, the risk factors regarding Hp infection and ITP through a uniformed questionnaire. Patients with Hp infection were diagnosed by combined detection of serum Hp antibodies and stool antigens. CagA and VacA proteins were tested by ELISA method. In addition, clinical data and follow-up data of aITP children were collected. Non-conditional logistic regression and t test were applied for statistical analysis. RESULTS: (1) The prevalence of Hp infection in aITP children and controls were 41.30% and 35.71%, respectively. No association between Hp infection and children aITP was found with OR of 1.170 (95%CI: 0.7163 - 1.673) after adjusting for confounding variables. (2) No statistical differences regarding initial platelet counts, megakaryocytes counts and the constituent ratio were found between the aITP children with and without Hp infection (P > 0.05). (3) No differences regarding initial platelet counts were found between aITP children with and without the expression of CagA (P > 0.05). The follow-up data showed that 32.88% of aITP children with Hp infection, as well as 29.70% of aITP children without Hp infection developed into cITP. No association between Hp infection and development to cITP was found with adjusted OR 1.171 (95%CI: 0.555 - 2.11 2). CONCLUSIONS: The results didn't suggest that Hp is unlikely to play a role in the onset of childhood aITP, and in the development of aITP to cITP.

[Association between platelet-activating factor acetylhydrolase gene polymorphism and intracranial hemorrhage in preterm infants].

OBJECTIVE: To explore whether Val279Phe single nucleotide polymorphisms (SNPs) in the 9th exon of platelet-activating factor acetylhydrolase (PAF-AH) are associated with intracranial hemorrhage in preterm infants. METHODS: A case-control study was performed. Polymerase chain reaction (PCR) was used to test genotype and allele frequencies of the 9th exon Val279Phe SNPs of PAF-AH in 58 preterm infants with intracranial hemorrhage (hemorrhage group) and 65 preterm infants without intracranial hemorrhage (control group). RESULTS: There were significant differences in genotype frequency of Val279Phe SNPs in the 9th exon of PAF-AH between the hemorrhage and control groups (P<0.05). Frequency of normal genotype in the hemorrhage group (63.8%) was significantly lower than in the control group (81.5%). In contrast, frequency of heterozygous genotype (34.5%) in the hemorrhage group was significantly higher than in control group (16.9%). There were also significant differences in allele frequency of Val279Phe SNPs in the 9th exon of PAF-AH between the two groups (P<0.05). T allele frequency in the hemorrhage group (19.0%) was significantly higher than in the control group (10.0%). CONCLUSIONS: Val279Phe SNPs in the 9th exon of PAF-AH may be associated with intracranial hemorrhage in preterm infants.

[Efficacy of immunosuppressive therapy for children with aplastic anemia].

OBJECTIVE: To study the effectiveness and safety of immunosuppressive therapy (IST) in the treatment of childhood aplastic anemia (AA) and to study the main factors influencing the effectiveness. METHODS: The clinical data of 55 children with severe aplastic anemia (SAA) and 51 children with chronic aplastic anemia (CAA) were retrospectively analyzed. All patients received IST from January 2007 to December 2010. RESULTS: In children with CAA, the effective rate of antithymocyte globulin (ATG) plus cyclosporine A(CsA) combination therapy was significantly higher than that of CsA alone (80% vs 44%; P<0.05); in children with SAA, the effective rate of ATG plus CsA combination therapy was also significantly higher than that of CsA alone (75% vs 40%; P<0.05). No patients developed clonal disease such as myelodysplastic syndrome, paroxysmal nocturn hemoglobinuria or acute myelocytic leukemia. In patients treated with the ATG plus CsA combination therapy, the response rate was relatively high for children whose disease course was less than six months, bone marrow hematopoietic area was more than 40%, had no severe infections, and experienced granulocyte colony stimulating factor (G-CSF) reaction during the early treatment; however, it was not related to AA subtypes and age. CONCLUSIONS: ATG plus CsA combination therapy is effective and safe in the treatment of childhood AA. The disease course, bone marrow hematopoietic area, severe infections and G-CSF reaction to early treatment are the main factors influencing the therapeutic effects.

[Clinical efficacy of desmopressin in the treatment of mild hemophilia A in children].

OBJECTIVE: To study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A. METHODS: Eighteen children with mild hemophilia A were enrolled. DDAVP (0.3 µg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment. RESULTS: In 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment. CONCLUSIONS: DDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.

[Cell proliferation and signal pathway after knockdown and RESC concurrent rescue of RNAi lentiviral vector on human PTEN gene in T-lymphocytes].

OBJECTIVE: To construct the lentiviral expression vectors of human PTEN gene for RNA interference (RNAi) and concurrent rescue of RNAi escape strategy construct (RESC) and to observe the changes of signal pathway, cell proliferation and cell cycle after PTEN gene knockdown and RESC concurrent rescue in human T-lymphocytes, in order to provide an experimental basis for a further research into the pathogenesis of acute lymphoblastic leukemia in children. METHODS: Using lentiviral vector systems to construct lentiviral vectors of human PTEN gene for RNAi and its RESC concurrent rescue, human T-lymphocytes were transfected with the lentiviruses. The cell models were established with PTEN gene knockdown (T-LC-shPTEN) and RESC concurrent rescue (T-LC-rrshPTEN). After knockdown and RESC concurrent rescue of PTEN gene, the expression of PTEN protein and the activation of AKT signal pathway, cell proliferation and cell cycle were detected by Western blot, MTT assay and flow cytometry respectively. RESULTS: The RNAi-mediated lentiviruses can down-regulate the expression of the human PTEN gene effectively. After the down-regulation of PTEN gene, the T-lymphocytes grew faster. The phase G0/G1 cells decreased and the phases S and G2/M cells increased significantly. The PI3K/AKT signal pathway was activated. All RNAi phenomenon caused by PTEN gene knockdown were recovered fully by RESC concurrent rescue of RNAi. CONCLUSIONS: The lentiviral expression vectors of human PTEN gene for RNAi and RESC concurrent rescue of RNAi are constructed successfully. The PI3K/AKT signal pathway can be activated and the proliferation of human T-lymphocytes can be promoted after PTEN gene knockdown.