Immune thrombocytopenic purpura is the common hemorrhagic disease in childhood.According to the international working group consensus,the article reviews its definition,classification criteria and the clinical features and diagnosis,in regarding the pathogenesis of immune function abnormalities,infection,platelets apoptosis and genetic background,espeially in children's treatment.

The experiment was designed to simulate active training with animal model. A dynamic and static trai-ning program for adult rats was carried out 30-60'/day, lasted from 25 to 232 days. The rats were killedperiodical1y in batches and the sections were prepared for exploring histological changes in knee and ankle areas on L/M. Degenerative cbanges in quadriceps and gastrocanemius muscles appeated by degrees aftet a period of in-tensive training. The nuclei proliferated in chains or clusters arranged in the center and the edge of thefibers. The number of nuclei for each chain or cluster could be as much as 20 or 30. Meanwhile, therewere also ischemic changes in small vessels. All these were the characteristic changes induced by mictotraumain muscles under repeated active and strenous training. In calcaneal and patellar ateas, there were changes of cartilaginous degeneration, necrosis and prolifera-tion (cartilaginous exostoses and osteophytes) too. The typical changes of enthesiopatby---degeneration, necrosis and proliferation (osteophyte) could beobserved in calcaneal tendon and its insertion area and in superior insertion atea of lower patella. The occur-rence of enthesiopathy in this experiment migbt be related to tke stress of traction force within muscle andcartilage during running in upward treadmill.

Objective To observe the clinical effects of eyehd reconstruction by regional flap combined with xenogeneic acellular dermal matrix transplantation for eyelid defect after malignant tumor excision.Methods Thirty-five cases (35 eyes) in our hospital were selected as the objects.Among them,basal cell carcinoma was 21 cases,meibomian gland carcinoma was 13 cases,squamous cell carcinoma was 1 cases;12 cases of upper eyelid and 23 case of lower eyelid were involved.All patients were subjected to intraoperative frozen,and the incision margin was determined according to the frozen results.After resection of the tumor,the eyelid had full-thickness defects in different degrees.The xenogenic acellular dermal matrix was used to replace the conjunctival tarsal tissue,and the adjacent flap or transposition flap was used to repair the eyelid defect according to the size of skin defect.The healing of flap,oral repair film,eyelid closure and adhesion were observed.Results After half a year follow-up,the acellular dermal matrix were completely dissolved by crawling the conjunctival epithelium covering,the flap healed with no flap necrosis.28 patients recovered well after operation without hypophasis and entropion,ectropion.4 patients had mild hypophasis,and there was no case of exposure keratitis.3 patients were with mild symblepharon.Conclusion The acellular dermal matrix can replace tarsal conjunctival tissue,which combined with regional flap has good clinical curative effect for eyelid defect after malignant tumor excision.This treatment can reduce the pain of patients who take oral mucosa and avoid the reoperation of eyelid reconstruction.

Objective To explore the initial therapy indications of acute immune thrombocytopenia (ITP) in children based on the classification treatment.Methods Three hundred and eighty newly diagnosed ITP cases were enrolled in this study from Jan.1st 2012 to Apr.30th 2013 in Children's Hospital Affiliated to Shanghai Jiaotong University.In total 380 patients,there were 214 male cases (56.31％) and 166 female cases (43.68％).The cases were divided into observation group and therapy group according to the initial platelet count which was ≤ 30 × 109/L or the bleeding over moderate volume or with active bleeding.Platelet values were observed in the observation group weekly,adrenal cortical hormone and immunoglobulin treatment were adopted in the therapy group,cases were followed up to Aug.30th 2013,9 months on the average.Results Three hundred and five cases showed overall response (80.26％) and 75 cases showed no response(19.74％).One hundred and seventy-eight cases were divided into observation group (46.84％),in which 133 cases (74.72％) showed complete response or response.Two hundred and two cases were included in therapy group (53.16％),in which 167 cases (82.67％) were with complete response or response.There was no statistical difference between the 2 groups in curative effect (Z =-0.54,P =0.59).Forty-five cases in observation group were no response and accepted therapy,35 of them (77.78％) had response.There were equal efficiency in the initial therapy group and the subsequent therapy group (x2 =3.60,P =0.06).There was no difference between the age of onset,sex and season in 2 groups.Cases aged from 1 month to 1 year seemed to have a high incidence because of vaccination,and in the cases aged from 3 to 14 years the onset was related to infection.The children over 3 years old had higher risk factors in self-healing and the curative effect.There was no severe bleeding or adverse effect or dead cases in this study.Conclusions It is feasible to take platelet count ≤30 × 109/L as the threshold for initial therapy indications.Almost half of the cases could avoid overtreatment and pretherapy observation will not reduce the initial cure effect; no severe internal bleeding was observed in all the cases.

OBJECTIVE:To establish a method for content determination of cyproheptadine hydrochloride in fuyan cream.METHODS:Ultraviolet spectophotometry was applied to determine the contents of cyprohepatadine hydrochloride as well as its adjuvant as two coexisting components without isolation and extraction,the wavelengths were286nm and258nm res_ pectively,linear regression and simultaneous equations were used to analyze the results.RESULTS:Cyproheptadine hydrochlor_ ide and its adjuvant showed good linear relationship in the range of2.52～25.2and2.5022～25.022?g/ml respectively,the average recovery of cyproheptadine hydrochloride was98.61%(RSD=1.08%).CONCLUSION:The present method is simple,convenient,reproducible and reliable,which is suitable for rapid determination for fuyan cream.

Objective:To differentiate human embryonic stem cells ( hESCs ) into keratinocytes ( K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs.Methods: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium.The hESCs were directly differentiated into kerati-nocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 sup-plement.The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR.Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques.Results:H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium.The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT ( P<0.05) , but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs ( P >0.05 ) .Conclusion: H9-hESCs could be directly differentiated into K-hESCs.The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTM). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTM were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-?) and interleukin-6(IL-6) in the hTM was analyzed by radioimmunoassay, after hTM were cultured with 0.312,0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTM induced by PPM, and the levels of TNF-? and IL-6 in the hTM induced by PPM increased markedly too. And the induced expression effect of TNF-? and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTM,and activate hTM in vitro.

Objective To sudy the changes in mTOR signaling pathway in childhood aplastic anemia(AA) by detecting the expression levels of the molecules of mTOR signaling pathway in T cells,and to explore immunologoical pathogenesis of AA in children from T cell intracellular signal transduction pathway.Methods Peripheral blood samples were collected from 16 newly diagnosed severe AA(SAA) patients and 8 patiens treated with effective immunosuppressive therapy,and the findings were compared with those of 17 healthy children (normal controls) and CEM cells (positive controls).The expressions of p-Akt,p-TSC2,p-mTORC1,p-4EBP1,p-p70S6K in CD3 + T cells in peripheral blood were detected by flow cytometry(FCM).Results 1.The expressions of p-Akt,p-TSC2,p-mTORC1,p-4EBP1,pp70S6K of the newly diagnosed SAA group were higher than those of the normal control group (P ＜ 0.05),but were lower than the postive control group (CEM group) (P ＜ 0.05).The mean fluorescence intensity (MFI) of p-Akt of three groups was 8.04 ± 3.78,2.59 ± 1.01 and 20.23 ± 8.98 respectively ;p-TSC2 was 49.73 ± 19.49,16.10 ± 8.04 and 101.05 ± 29.78 respectively ; p-mTOR was 13.90 ± 9.32,2.92 ± 1.09 and 34.3 ± 19.03 ;p-4EBP1 was 142.69 ± 53.36,26.91 ± 13.70,256.01 ± 53.79 ; p-p70S6 K were 17.67 ± 10.48,3.69 ± 2.22,31.73 ± 12.85 respectively.2.The expressions of p-Akt,p-TSC2,p-mTORC1,p-4EBP1,p-p70S6K of the effective treatment groups were lower than those of the newly diagnosed SAA group (P ＜ 0.05) ; the expressions of p-Akt,p-TSC2,p-mTORC1,p-p70S6K were similar to those of the normal control group(P ＞ 0.05),but the expressions of p-4EBP1 were higher(P ＜ 0.05).The MFI was followed by 3.28 ± 1.27,16.50 ± 10.91,3.54 ± 1.66,74.89 ± 49.69 and 4.21 ± 1.69.Conclusions 1.The expressions of p-Akt,p-TSC2,p-mTORC1,p-4EBP1,p-p70S6K were increased in the newly diagnosed SAA patients,the mTOR signaling pathway was activated in SAA patients.2.The expressions of p-Akt,p-TSC2,p-mTORC1,p4EBP1,p-p70S6K were lower than those of the newly diagnosed SAA patients.The degree of activation of mTOR signaling pathway was associated with disease status.The signaling pathways may be involved in the T cells of AA of the immune abnormalities.

Objective To investigate the effects of high-fat diet induced insulin resistance on fibroblast growth factor-21 (FGF-21) and its receptors expression in ApoE~(-/-) mice. Method Male ApoE~(-/-) mice were randomly divided into normal-chow group(NF,n=20)and high-fat fed group(HF,n=20) and fed for 16 weeks. The insulin sensitivity and glucose-lipid metabolism in awake mice were evaluated by hyperinsulinemic-euglycemic clamp technique combined with 3-[~3H]-glucose as a tracer. The Mrna expressions of FGF-21,β-klotho, and FGFR1-4 were measured by quantitative real-time PCR. FGF-21 protein levels were determined by Western blot. Results Fasting blood glucose, plasma insulin and free fatty acids, triglycerides, free fatty acids, and cholesterols were significantly elevated in HF group compared with NF group(all P<0.01). During the steady-state of clamp, plasma insulin was significantly higher in HF group than that in NF group(P<0.01), and glucose infusion rate was also significantly decreased(P<0.01). At the end of insulin clamp, glucose disappearance rate was significantly lower in HF group than that in NF groups(P<0.01). Hepatic glucose production in NF group was suppressed by 70% ,while in HF group it was suppressed by 51%. The FGF-21 Mrna expressions of hepatic and adipose tissues in HF group were significantly increased compared with NF group(both P<0.01), and β-klotho Mrna expressions increased(P<0. 05). In hepatic and adipose tissues, FGFRI, Mrna expressions were higher in HF group than those in NF group(both P<0.01) ,and FGFR3 Mrna increased(P<0.01 and P<0.05, respectively). In hepatic tissue,FGFR4 Mrna levels were significantly up-regulated in HF group(P<0. 05). Plasma FGF-21 levels were elevated in HF group compared with NF group(P<0.01) ,and FGF-21 protein expressions of hepatic and adipose tissues were also increased(both P<0.05). Conclusion FGF-21, β-klotho, FGFR1, and FGFR3 were significantly up-regulated in ApoE~(-/-) mice fed by high-fat diet, and they might be the targets in regulating glucose-lipid metabolism by FGF-21.

To synthesize salbutamol immunogen and develop an enzyme immunoassay (ELISA), a new salbutamol immunogen was synthesized using 4-aminobenzoic acid as a linker to connect hapten with carrier protein. An enzyme immunoassay based on the antibody prepared was developed and applied to detect salbutamol residue spiked in swine liver. An unusual coating antigen, clenbuterol-ovalbumin (OVA) conjugate instead of salbutamol-OVA conjugate, was used in the immunoassay and the results were discussed based on the structures of related compounds. The antibodies showed high sensitivity in the heterologous assay when using clenbuterol-OVA as a coating antigen, with an IC50 value of 8.97 ng mL(-1) toward salbutamol. The antibodies prepared showed high cross-reactivity with clenbuterol (107%) and were promising for the simultaneous determination of salbutamol and clenbuterol residues in food and food products. Recovery rates from the salbutamol-spiked swine liver samples were in the range of 70%-99%, while the intra-assay and inter-assay coefficients of variation were <13.3% and <14.3%, respectively. In summary, the antibodies of salbutamol have been successfully prepared. Sensitive and stable analysis for the detection of salbutamol residues in swine liver was obtained based on the competitive ELISA methods developed in this study.