[Effect of ultraviolet irradiation on the proliferation of acute promyelocytic leukemia cells under hypoxic conditions and related mechanisms].

OBJECTIVE: To study the effect of 280 nm-LED ultraviolet irradiation on the proliferation of acute promyelocytic leukemia (APL) HL-60 cells under hypoxic conditions and related mechanism. METHODS: HL-60 cells in the logarithmic growth phase were selected and divided into control, hypoxia, ultraviolet and hypoxia+ultraviolet groups. The cells in the hypoxia group were treated with cobalt chloride (with a final concentration of 150 µmol/L), those in the ultraviolet group were irradiated by 280 nm-LED ultraviolet with an energy intensity of 30 J/m2, and those in the hypoxia+ultraviolet group were treated with cobalt chloride and then irradiated by 280 nm-LED ultraviolet. After 48 hours of treatment, the cells were placed under an invert microscope to observe cell morphology. CCK-8 assay was used to measure the inhibition rate of cell proliferation. Annexin V-FITC/PI double staining flow cytometry was used to evaluate cell apoptosis. Quantitative real-time PCR was used to measure the mRNA expression of Bcl-2. Each experiment above was repeated three times independently. RESULTS: Compared with the control group, the experimental groups showed shrinkage, decreased brightness, and disordered arrangement of cells, and the number of cells decreased over the time of culture. There were significant differences in the inhibition rate of cell proliferation and cell apoptosis rate among the groups (P<0.01), and the hypoxia+ultraviolet group showed the strongest inhibition of cell proliferation and induction of cell apoptosis, followed by the ultraviolet group and the hypoxia group. Compared with the control group, the other three groups had a gradual reduction in the mRNA expression of Bcl-2, and the hypoxia+ultraviolet group had a significantly greater reduction than the hypoxia and ultraviolet groups (P<0.01). CONCLUSIONS: Both hypoxia and ultraviolet irradiation can inhibit the proliferation of HL-60 cells and induce cell apoptosis, and ultraviolet irradiation has a better effect on proliferation inhibition and cell apoptosis under hypoxic conditions than under normoxic conditions, possibly by downregulating the mRNA expression of Bcl-2.

Clinical presentation and outcome of pediatric patients with hemophagocytic lymphohistiocytosis in China: A retrospective multicenter study.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a heterogeneous disease with major diagnostic and therapeutic difficulties. A large-scale multicenter study of pediatric HLH is still lacking in China. PROCEDURE: The Histiocytosis Study Group of the Chinese Pediatric Society conducted this retrospective study in 2014. A total of 323 patients diagnosed with HLH between 2011 and 2013 from 12 hospitals were registered. RESULTS: The median age at diagnosis was 2.2 years (range, 0-14.6 years), with a peak age of HLH onset at 0 to 3 years (63%). Mutations in HLH-related genes were found in 27.9% (24/86) patients who underwent genetic testing. PRF1, UNC13D, STXBP2 and LYST were the predominant genes involved. Sixteen patients (66.7%) presented with only monoallelic mutations in one gene. Epstein-Barr virus (EBV) infection was the major condition related to HLH, which was documented in 74.4% (201/270) of the patients who underwent EBV detection. Of 252 evaluable patients, 64.7% (163) achieved non-active disease at the eighth week and patients treated with a protocol containing etoposide presented higher remission rates (75.6% vs. 46.8%, P < 0.001). In multivariate analysis, a younger age at diagnosis (<12 months), platelet count less than 80×109 /L, central nervous system involvement, and initial treatment using a protocol without etoposide (not HLH-94/04) were independent prognostic factors indicating resistant disease. DISCUSSION: This study first multicenter assessment of HLH in China shows some different features in Chinese children with HLH compared with those in western countries, including older age, vulnerability to EBV infection, and a high proportion of patients with single monoallelic genetic mutations.

Purification and Identification of Antioxidant Peptides from Protein Hydrolysate of Scalloped Hammerhead (Sphyrna lewini) Cartilage.

The aim of this study was to purify and identify peptides with antioxidant properties from protein hydrolysate of scalloped hammerhead (Sphyrna lewini) cartilage. Cartilaginous proteins of the scalloped hammerhead were extracted by guanidine hydrochloride, and three antioxidant peptides, named enzymolysis peptide of scalloped hammerhead cartilage A (SCPE-A), SCPE-B and SCPE-C, were subsequently isolated from the hydrolysate of the cartilaginous proteins using ultrafiltration and chromatography. The amino acid sequences of SCPE-A, SCPE-B and SCPE-C were identified as Gly-Pro-Glu (GPE), Gly-Ala-Arg-Gly-Pro-Gln (GARGPQ), and Gly-Phe-Thr-Gly-Pro-Pro-Gly-Phe-Asn-Gly (GFTGPPGFNG), with molecular weights of 301.30 Da, 584.64 Da and 950.03 Da, respectively. As per in vitro activity testing, SCPE-A, SCPE-B and SCPE-C exhibited strong scavenging activities on 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH•) (half elimination ratio (EC50) 2.43, 2.66 and 1.99 mg/mL), hydroxyl radicals (HO•) (EC50 0.28, 0.21 and 0.15 mg/mL), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS⁺•) (EC50 0.24, 0.18 and 0.29 mg/mL), and superoxide anion radicals ( O 2 - •) (EC50 0.10, 0.14 and 0.11 mg/mL). In addition, SCPE-A showed inhibition activity similar to butylated hydroxytoluene (BHT) in lipid peroxidation in a linoleic acid model system. The amino acid residues of Gly, Pro and Phe could positively influence the antioxidant activities of GPE, GARGPQ and GFTGPPGFNG. These results suggested that GPE, GARGPQ and GFTGPPGFNG might serve as potential antioxidants and be used as food additives and functional foods.

[Expression of PTEN in Myocardial Tissue in Coronary Heart Disease].

OBJECTIVE: To observe the expression of phosphatase and tensin homology deleted on chromosome ten (PTEN) in myocardial tissue in patients with coronary heart disease, and explore the relevance between the expression of PTEN and the occurrence and development of coronary heart disease. METHODS: A total of 16 death cases with pathological diagnosis of coronary heart disease were collected as experimental group, and 19 cases without myocardial lesions were selected as control group. The expression of PTEN protein and its mRNA were detected by immunohistochemistry and real-time fluorescence quantitative PCR respectively. The correlation between the expression of PTEN and the pathogenesis of coronary heart disease was analyzed. RESULTS: The expression of PTEN protein in myocardium in cases with coronary heart disease was significantly lower compared with the control group (P < 0.05). There was no statistical difference of the expression of PTEN mRNA between experimental and control group (P > 0.05). CONCLUSION: PTEN may be involved in the occurrence and development of coronary heart disease.

[Relationship between clinical features and cognitive function in patients with childhood and adolescence-onset schizophrenia].

OBJECTIVE: To explore the factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia. METHODS: The clinical data of 78 patients with childhood and adolescence-onset schizophrenia who met with the criteria of ICD-10 for schizophrenia were retrospectively reviewed. The cognitive functions were evaluated by the Chinese Wechsler Intelligence Scale for Children (C-WISC), the Wisconsin Card Sorting Test (WCST), digit span backward and P300. The clinical symptoms were evaluated by the Positive and Negative Syndrome Scale (PANSS). RESULTS: The patients with a lower education level or earlier onset of age had a longer P3 latency at the P300Fz area. The patients with a higher parental education level had higher scores of full intelligence quotient (FIQ), verbal intelligence quotient (VIQ), performance intelligence quotient (PIQ), conceptual level and completed categories of WCST and backward numeric order reciting. The patients with higher PANSS negative subscale scores had lower scores of FIQ, VIQ, PIQ, completed categories and conceptual level of WCST and backward numeric order reciting. The patients with a longer stabilization time had higher backward numeric order reciting scores. CONCLUSIONS: The severity of negative symptoms of the patients and the educational level of their parents are major factors influencing cognitive functions in patients with childhood and adolescence-onset schizophrenia.

[Construction of Plasmodium falciparum signal peptide peptidase-GFP mutant and its expression analysis in the malaria parasite].

OBJECTIVE: To construct recombinant plasmid pSPPcGT which contains signal peptide peptidase gene of Plasmodium falciparum (PJSPP) and GFP, and transfect into P. falciparum (3D7 strain) to obtain mutant parasites which can express PJSPP-GFP. METHODS: Plasmodium falciparum(3D7 strain) genomic DNA was extracted from cultured malaria parasites. The C-terminal region of PJSPP, an 883 bp gene fragment was amplified by PCR, and then cloned into pPM2GT vector to get recombinant vector pSPPcGT. The recombinant vectors were identified by PCR, double restriction enzyme digestion and DNA sequencing. pSPPcGT vector was transfected into malaria parasites. The positive clones were selected by adding inhibitor of Plasmodium falciparum dihydrofolate reductase WR99210 to the culture medium. The pSPP-GFP-transfected parasites were fixed with methanol, stained with DAPI, and observed under immunofluorescence microscope. The PJSPP-GFP expression in P. falciparum was identified by SDS-PAGE and Western blotting. RESULTS: The C-terminal region of PJSPP was amplified from P.falciparum (3D7 strain) genomic DNA by PCR with the length of 883 bp. The constructed recombinant vectors were identified by PCR screening, double restriction enzyme digestion and DNA sequencing. The pSPPcGT vector was transfected into P. falciparum and the positive clones were selected by WR99210. GFP fluorescence was observed in transfected parasites by immunofluorescence microscopy, and mainly located in the cytoplasm. The PJSPP-GFP expression in malaria parasites was confirmed by Western blotting with a relative molecular mass of Mr 64,000. CONCLUSION: Recombinant vector PJSPP-GFP is constructed and transfected into P. falciparum to obtain P. falciparum mutant clone which can express PfSPP-GFP.

[Effect of parent training in combination with methylphenidate treatment on family relationships for children with attention deficit/hyperactivity disorder].

OBJECTIVE: To investigate the effect of parent training combined with methylphenidate treatment on family relationships in children with attention deficit/hyperactivity disorder (ADHD). METHODS: Fifty-nine parents of children with ADHD under methylphenidate treatment participated in a modified 5-week training program. The intervention effect was evaluated using the Conners Parent Symptom Questionnaire, ADHD Rating Scale-IV Home Version (ADHD-RS-IV Home Version), Caregiver Strain Questionnaire, Parent-Child Relationship Self-rating Scale and Piers-Harris Children's Self-Concept Scale. Parents also completed the training satisfaction survey before and after the intervention. RESULTS: After the 5-week parent training, compared with the baseline values, total scores of Conners Parent Symptom Questionnaire and scores of conduct problems and anxiety significantly decreased, and scores of attention deficit, hyperactivity, impulsivity and oppositional defiant behaviors of ADHD-RS-IV Home Version, and Caregiver Strain Questionnaire total scores were all significantly decreased (P<0.05), while total scores of the Parent-Child Relationship Self-Rating Scale and Piers-Harris Children's Self-Concept Scale were significantly increased (P<0.05). CONCLUSIONS: Modified 5-week parent training program may improve parent-child relationship and reduce parenting stress in ADHD families.

[Study on attention-deficit hyperactivity disorder in children based on resting-state functional magnetic resonance imaging].

OBJECTIVE: To compare resting-state functional magnetic resonance imaging (fMRI) findings of children with attention-deficit hyperactivity disorder (ADHD) and normal children, and to investigate the possible mechanism of brain dysfunction in children with ADHD. METHODS: Resting-state fMRI was performed on 18 children who met the DSM-IV diagnostic criteria for ADHD (ADHD group) and 18 normal children (control group) matched for age, sex, IQ, degree of education and handedness. The two groups were compared in terms of amplitude of low frequency fluctuation (ALFF) and regional homogeneity (ReHo). RESULTS: Compared with the control group, the ADHD group had decreased ALFF in the bilateral posterior lobes of the cerebellum and the left side of the pons, increased ALFF in the right precentral gyrus, decreased ReHo in the left medial frontal gyrus, right superior frontal gyrus, and left precuneus, and increased ReHo in the left anterior lobe of the cerebellum, left caudate nucleus, right parahippocampal gyrus, left precentral gyrus, and right middle frontal gyrus. CONCLUSIONS: In resting state, children with ADHD have decreased brain activity in some regions, including the cerebellum and frontal cortex, compared with normal children, which supports the hypothesis of dysfunctional fronto-cerebellar circuits in ADHD.

[Influential factors for methylphenidate treatment compliance in children with attention deficit hyperactivity disorder].

OBJECTIVE: To investigate influential factors for the tendency to medicate and medication compliance in children with attention deficit hyperactivity disorder (ADHD). METHODS: A total of 188 children aged from 5 to 16 years, who were initially diagnosed with ADHD according to DSM-IV criteria, were included in the study. They underwent symptom assessment and cognitive function test. The compliance of methylphenidate treatment was evaluated. RESULTS: Patients with better emotional state, and fewer oppositional and hyperactive behaviors and those who had a family history of psychiatric diseases and who obtained lower scores in the number cancellation test (NCT), were more prone to medication and/or exhibited better medication compliance. Logistic regression analysis showed that fewer oppositional and hyperactive behaviors and lower NCT scores were the predictive factors for a higher tendency to medicate, and a better emotional state was the predictive factor for better medication compliance. Patients of predominantly inattentive type were more prone to medication and showed better medication compliance, as compared with those of combined type. Gender, age and symptom severity were not associated with the tendency to medicate and/or medication compliance. CONCLUSIONS: There is a need to enhance medication compliance in children with ADHD who have hyperactive, impulsive and oppositional behaviors, and to improve their long-term social functions.

Antioxidant Peptides From Protein Hydrolysate of Marine Red Algae Eucheuma cottonii: Preparation, Identification, and Cytoprotective Mechanisms on H2O2 Oxidative Damaged HUVECs.

To screen, prepare, identify, and evaluate the activities of natural antioxidants for treating chronic diseases caused by oxidative stress. Two algal proteins, namely ZD10 and ZD60, precipitated with 10 and 60% (NH4)2SO4 were extracted from red algae Eucheuma cottonii (E. cottonii) and hydrolyzed using five proteolytic enzymes. The results showed that ZD60 played the most significant role in the enhancement of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH⋅) scavenging activity (25.91 ± 0.24%) among all protein hydrolysates. Subsequently, six antioxidant peptides (EP1-EP6) were isolated from the papain hydrolysate of ZD60 by ultrafiltration and chromatography methods. Their amino acid sequences were identified as Thr-Ala (EP1), Met-Asn (EP2), Tyr-Ser-Lys-Thr (EP3), Tyr-Ala-Val-Thr (EP4), Tyr-Leu-Leu (EP5), and Phe-Tyr-Lys-Ala (EP6) with molecular weights of 190.21, 263.33, 497.55, 452.51, 407.51, and 527.62 Da, respectively. Of which, EP3, EP4, EP5, and EP6 showed strong scavenging activities on DPPH⋅, hydroxyl radical (HO⋅), and superoxide anion radical (O- 2⋅). Moreover, EP4 and EP5 could significantly protect human umbilical vein endothelial cells (HUVECs) from H2O2-induced oxidative damage by increasing the levels of antioxidant enzyme systems including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to reduce the levels of reactive oxygen species (ROS) (60.51 and 51.74% of model group) and malondialdehyde (MDA) (75.36 and 64.45% of model group). In addition, EP4 and EP5 could effectively inhibit H2O2-induced apoptosis by preventing HUVECs from early apoptosis to late apoptosis. These results indicated that the antioxidant peptides derived from E. cottonii, especially EP4 and EP5, could serve as the natural antioxidants applied in pharmaceutical products to treat chronic cardiovascular diseases caused by oxidative damage, such as coronary heart disease, atherosclerosis, etc.