Objective To investigate the effect of miRNA -1 00 on the proliferation of human leukemia cells HL -60,and to explore the mechanism of this action.Methods The bioinformatics software and database were applied to predict and analyze target genes of miRNA -1 00.The vector contained the target gene 3′UTR portion cloned into a luciferase reporter construct.A luciferase reporter assay was performed following co -transfection of small molecular miRNA -1 00 mimics and target gene wild -type or mutant plasmid into HEK -293T cells.HL -60 cells were trans-fected with miRNA -1 00 mimics or anti -miRNA -1 00.After transfection,Western blot was applied to validate the expression of carboxy -terminal domain small phosphatase -like protein (CTDSPL),and the viability of HL -60 was measured by using cell counting kit (CCK -8)assay at 24 h,48 h,72 h,96 h.Results Online software predicted that CTDSPL was likely to be the target gene of miRNA -1 00.Dual luciferase reporter gene assay system showed that miRNA -1 00 could significantly suppress the activity of reporter gene containing CTDSPL 3′-UTR which decreased by about 57.1 %(P =0.000 7).Western blot showed that the expression of CTDSPL was increased after being trans-fected with miRNA -1 00 antisense oligonucleotides and decreased after being transfected with miRNA -1 00 mimics.At the same time,the growth rate of cells treated with miRNA -1 00 mimics or CTDSPL miRNA -1 00 was increased com-pared with that in control by CCK -8 test (P <0.05 ).Conclusions CTDSPL is a downstream target gene of miRNA -1 00.miRNA -1 00 can promote leukemia cell proliferation by inhibiting the expression of CTDSPL.

Objective To explore the effect of Flavokawain B on the proliferation and apoptosis of acute T lymphoblastic leukemia(T -ALL)cells and its preliminary mechanism.Methods After the T -ALL cell lines CEM-C7(sensitive to glucocorticoids)and MOLT -4(resistant to glucocorticoids)cells were treated with different concentrations of Flavokawain B,the influence of Flavokawain B on the growth rate and doubling time of CEM-C7 and MOLT -4 cells was observed by 3 -(4,5 -dimethylthiazol -2 -yl)-5 -(3 -carboxymethoxyphenyl)-2 -(4 -sulfophenyl)-2H -tetrazolium(MTS)assay,and apoptosis was analyzed by using flow cytometry.Furthermore,Wes-tern blot assay was used to detect the expressions of Bim,Bcl -2 and cleaved Caspase -9.At last,the expressions of Bim and Bcl -2 in clinical T -ALL patient samples were also detected by using Western blot assay.Results MTS as-say showed that Flavokawain B significantly inhibited the cellular proliferation of T -ALL cell lines in a dose and time dependent manner(P <0.01 ).Flow cytometry findings revealed that Flavokawain B significantly induced the apoptosis of T -ALL cells in a dose -dependent manner(P <0.001 ).Western blot results indicated that Flavokawain B in-creased the expression of Bim and cleaved Caspase -9,and decreased the expression of Bcl -2 in T -ALL cell lines, which increased Bim and decreased Bcl -2 in clinical T -ALL patients samples,both in a dose -dependent manner. Conclusions Flavokawain B can inhibit the proliferation and induce the apoptosis of T -ALL cells by up -regulating the expression of Bim and down -regulating the expression of Bcl -2 and activating Caspase -9,whether resistant to glu-cocorticoids or not.

Hepatoblastoma (HB)is the most common malignant liver tumor in children.Surgical resection was the major treatment approach to HB,whereas more than 60% patients had unresectable mass at diagnosis,even though complete resection,there were about 30% patients recurrent.With the introduction of adjuvant chemotherapy,the treat-ment of HB has been developed to multidisciplinary cooperation from single surgical resection,such strategy greatly im-proved the prognosis of children with HB,but there still be a great challenge in the treatment of high -risk patients.All famous children′s HB study groups have begun to focus on the optimization of chemotherapy,surgery opportunity,strate-gy based on risk -stratification,looking for new potential therapeutic targets and prognosis -related risk factors.

Pediatric acute myeloid leukemia(AML)is still a fatal disease.Recent evidence based clinical study indicated intensified chemotherapy contribute to improvement of pediatric AML survival,while supporting care was an important measurement to let patients tolerate more and more intensifying chemotherapy.Recent series of clinical trial reveals that benefit from more intensified chemotherapy will be limited.Combined risk stratification of genetics and minimal residual desease monitoring may further increase the survival.Tyrosine kinase inhibitors,Gemtuzumab ozogami-cin and Chimeric antigen receptors modified T cell based targeted therapy could be the only way to cure resistant AML. In this review,advance in conventional chemotherapy,hematopoietic stem cell transplantation,and targeted therapy for pediatric AML were reviewed.

Group A β-hemolytic streptococcus (GAS),namely Streptococcus pyogenes,is one of the most im-portant human pathogen.GAS can cause skin and mucous membrane superficial infectious diseases,life -threatening invasive disease,toxin -mediated diseases and immune -related diseases.Antibiotic is an effective mean to control GAS infection.The β-lactam antibiotics remain the first -choice treatment for GAS infection and the macrolides are often recommended as a replacement therapy for β-lactam antibiotics allergic patients or a means to blocking GAS exotoxin product.But with the widespread use of macrolides autibiotics,macrolide -resistant GAS spread in the world. This paper will elaborate the situation of macrolide -resistant clones.

Antimicrobial resistance has been identified as one of the world′s most pressing public health threats.The number of antibiotic -resistant bacteria and the diversity of molecular mechanisms of resistance have in-creased sharply in recent years.The presence of antimicrobial resistance in children complicates patient management and arouses widespread concern.The use of antimicrobial agents is the single most important factor leading to the deve-lopment of resistance.New effective antimicrobial agents has been needed but has not kept pace.Antimicrobial -resis-tant infection can be prevented by immunization as well.Appropriate and judicious use of antimicrobial agents is the most important factors contributing to control resistance.Clinicians should collaborate well with microbiologist and know well about resistant bacteria and its proper antimicrobial drugs.A good application of antibiotic formulary restriction has the potential to decrease antimicrobial resistance.An antimicrobial resistance surveillance between regions and countries is urgently needed.It can help us to understand and slow antimicrobial resistance in the long run.

Objective To explore the psychological behavior features between different sexual children with oppositional defiant disorder(ODD).Methods The subjects were sampled from grade 1 to 5 in a primary school in Ji-nan,Shandong Province.According to the diagnosed criteria,42 pupils were diagnosed ODD,which was the study group,and 556 pupils without ODD became the control group.The pupils and their families were investigated and as-sessed by using Achenbach Child Behavior Checklist (CBCL).Results According to CBCL,there was extremely sig-nificantly higher score of depression,obsessive -compulsive,hyperactive,aggressive,interpersonal behaviors in ODD boys [(5.00 ±3.73)scores,(4.88 ±3.65)scores,(5.21 ±3.49)scores,(1 0.1 0 ±6.85)scores,(2.88 ±1 .99) scores]than those in healthy control group [(1 .65 ±2.61 )scores,(1 .91 ±2.65)scores,(2.46 ±2.63)scores, (3.95 ±4.52)scores,(1 .34 ±1 .61 )scores](t =4.31 ,3.90,4.76,4.33,4.40;P =0.000,0.000,0.001 ,0.000, 0.000).There was significantly higher score of symptoms of somatization,indiscipline,schizoid -like behaviors in ODD boys with [(2.38 ±2.50)scores,(2.63 ±2.70)scores,(3.1 7 ±2.35)scores]than those in healthy control group [(0.85 ±1 .63)scores,(0.1 3 ±1 .76)scores,(1 .51 ±1 .72)scores](t =2.94,2.85,3.37;P =0.007,0.009, 0.002).There was extremely significantly higher score of depression,communication disorder behaviors in ODD girls [(5.44 ±4.1 2)scores,(3.1 1 ±2.93)scores]than those in healthy control group with [(2.50 ±3.34)scores, (1 .36 ±2.08)scores](t =3.57,3.38;P =0.000,0.001 ).A significantly higher score of hyperactive,aggression, cruelty behaviors in ODD girls [(4.67 ±4.41 )scores,(9.33 ±7.82)scores,(1 .1 7 ±1 .79)scores]than those in healthy control group [(1 .87 ±2.38)scores,(3.1 9 ±4.08)scores,(0.24 ±0.66)scores](t =2.67,3.30,2.1 8;P =0.01 6,0.004,0.043).Conclusions According to CBCL,there were more psychological behavior problems in ODD children than healthy children.A significantly higher difference was found in different gender.

Objective To study the effect of bifidobacterium on intestinal tissue of necrotizing enterocolitis (NEC)in newborn rats and its regulation of Wnt/β-Catenin signal pathway.Methods Seventy -five newborn SD rats were randomly divided into 5 groups,and each group had 1 5 rats.Group A was artificial feeding control group;group B was NEC model group;group C was bifidobacterium treatment group;group D was artificial feeding +bifidobacterium control group;group E was rat breast feeding control group.The localization expression of Toll -like re-ceptor 4(TLR4)of ileocecal ileum tissue was detected by immunohistochemical detection,and also the equivalen-tileum tissues were detected for the contents of glycogen synthase kinase -3β(GSK3β)and β-Catenin expression by Wes-tern blot.Comparing the differences of these indicators between the groups,in addition,the data of TLR4,GSK3βandβ-Catenin were analyzed by Bivariate correlations.Results The levels of TLR4 in ileum tissue of 5 groups were 0.36 ±0.03,0.48 ±0.05,0.34 ±0.03,0.37 ±0.04,0.35 ±0.02.The levels of GSK3βin ileum tissue of 5 groups were 0.98 ±0.23,1 .48 ±0.42,0.99 ±0.20,0.56 ±0.1 7,0.60 ±0.1 5.The levels of β-Catenin in ileum tissue of 5 groups were 1 .48 ±0.22,0.64 ±0.55,1 .27 ±0.36,1 .72 ±0.51 ,1 .82 ±0.44.The levels of TLR4 and GSK3βin ileum tissue of group B were significantly increased compared with group E (P <0.05).The levels of β-Catenin sig-nificantly decreased compared with group E (P <0.05).The levels of TLR4 and GSK3βin ileum tissue of group C were significantly decreased compared with group B (P <0.05).The levels of β-Catenin significantly increased com-pared with group B (P <0.05).Negative correlation was observed between the levels of GSK3βand β-Catenin(r =-0.592,P <0.05),while positive correlation was observed between the levels of TLR4 and GSK3β(r =0.295,P <0.05),and negative correlation was observed between the levels of TLR4 and β-Catenin(r =-0.426,P <0.05). Conclusions Bifidobacterium has certain protective effect on the NEC newborn rat intestines,which can reduce the in-cidence of experimental NEC and the severity of intestinal injury.Its effect may be achieved by regulating the Wnt/β-Catenin signal pathway,which decreases the expression of the level of GSK3βand increases the level of repair fac-tor β-Catenin.

Objective To explore the clinical manifestations,imaging findings,pathological classification and treatment of congenital cystic adenomatoid malformation (CCAM)of the lung.Methods The clinical features,imaging findings,pathology information,diagnosis,treatment method and its prognosis of children with CCAMconfirmed by ope-ration and pathology were retrospectively analyzed in Yuying Children′s Hospital Affiliated to Wenzhou Medical Univer-sity from August 2006 to August 201 4.Results Eleven patients were boys and 4 patients were girls.One case had a-symptomatic clinical features,1 2 cases had pulmonary infection,1 case had recurrent chest pain,and 1 case had de-pressed deformity in sternum inferior segment.Chest CT scanning indicated that 9 cases had multiple gas cysts at unila-teral side of lung,among which 1 case was of funnel chest and pulmonary sequestration,1 case of huge cyst containing air and fluid at inferior lobe of left lung,and 4 cases of high density lung shadow;CT examination indicated that 1 case had recurrent chest pain and eventration of diaphragm of the right side combined with pulmonary sequestration.All ca-ses were treated by surgical resection,of whom 1 case was given cystectomy and sequestrectomy,diaphragmatic plication respectively,1 case complicated with funnel chest disease underwent lesion pulmonary lobectomy,sequestrectomy and minimally invasive corrective surgery in pectus excavatum (Nuss surgery),and the remaining 1 2 cases received lesion pulmonary lobectomy.All of 1 5 cases recovered well without complications.Pathological classification type of CCAMin-cluded 1 1 cases of type Ⅰ,3 cases of type Ⅱ and 1 case of type Ⅲ,among which 2 cases had pulmonary sequestration. Conclusions CCAMis a rare disease which can be discovered along with pulmonary infection.Multiple gas cysts are the most common imaging findings and the preoperative diagnosis of CCAM is mostly based on chest CT examination. Type Ⅰ and type Ⅱ are the most common pathological classification.The surgical resection should be given early surgi-cal resection and the prognosis is usually good.

Aplastic anemia (AA)is a serious disease of hematological malignant disease in children.Severe AA is difficult to treat and may lead to high motality.Acquired AA accounts for 95% of all cases,so it′s significant to lucubrate the pathogenesis of acquired AA for clinic diagnosis and therapy.The documents in recent 5 years are collected,and a reviews about the progress of pathogenesis in acquired AA children is provided.