Objective To investigate the effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial (RPE) cells and the signal transduction pathway. Methods Reverse transcription polymerase chain reaction and immunofluorescence staining were used to detect the expression of syndecan-1 mRNA and protein in normal RPE cells. The expression of syndecan-1 in RPE cells stimulated by different cytokines was detected and quantitatively analyzed by image process of immunofluorescence. The stimulation included 7 and 35 ng/ml tumor necrosis factor (TNF)-? for 24 hours, 1 and 6 ?g/ml lipopolysaccharide (LPS) for 11 hours, 7 ng/ml TNF-? for 0 to 24 hours (once per 2 hours, and 13 times in total), and 30% supernatant of monocyte/macrophage strain (THP-1 cells) for 3, 14 and 43 hours. The effect of 30% supernatant of THP-1 cells was assayed after pretreated by PD098059 [the specific inhibitor of extracellular signal regulated kinase(ERK) 1/2] for 2 hours. After exposed to 30% supernatant of THP-1 cells for 3 hours and treated by 0.25% trypsin for 5 minutes, RPE cells attaching was evaluated by methyl thiazolyl tetrazolium assay. Results In normal human RPE cells, expressions of syndecan-1 mRNA and protein were detected, and strong syndecan-1 positive yellowish green fluorescence was found in the cell membrane and cytoplasm while light green fluorescence was in the nucleus. As the concentration and stimulated time of TNF-? or LPS increased, the fluorescence intensity decreased(P

Objective To analyze, treatment of ipsilateral Pilon and calcaneal fractures (IPCF). Methods Injury causes included fallings in seven patients, traffic injury in two and impact injury in one, with ISS score of 5-22 points. There were seven patients with open injuries and 3 with close ones. Nine pa-tients had Ⅱ-Ⅲ degree of Pilon fractures according to the AO/OTA classification and six comminuted com-pression calcaneal fractures according to the Essex-Loprest classification. The Pilon fractures were fixed by cannulated nail, screw or Kirschner wire through open reduction in six patients including four fixed with ex-ternal fixators (the calcaneal fractures were fixed with percutaneous eannulated nail or Kirschner wire by poking reduction). The calcaneal fractures were fixed with only Kirschner wire through poking reduction in two patients and double fractures treated conservatively in two. Results The reduction of Pilon and cal-caneal fractures treated by operation were markedly improved in six patients, while there occurred angular deformity and joint surface introeession in the patients treated conservatively. The wound healing of soft tis-sue was sound in seven patients but unsatisfactory in the other three. The ankle-hind foot score was 82-94 points in four patients who were followed up. Conclusions IPCF is caused by the high energy force and characterized by severe injures of the soft tissues and the bone tissues at the distal tibia and calcaneua, for which limited internal fixation plus external fixation are suitable treatment choice.

OBJECTIVE To find the safer anesthesia induction methods in children with laryngeal papilloma combined by dyspnea.METHODS 50 laryngeal papilloma children with II degree laryngeal obstruction were randomly divided into 2 groups:inhalational group and intravenous group,no muscle relaxant was used in either of the groups.MAP,and SpO2 were observed at one minute before and after induction.The intubation condition was also assessed and compared between the 2 groups.RESULTS MAP and HR in intravenous group were significantly higher than those of inhaled group after intubation [(68.7?6.4)mmHg vs.(64.0?8.0)mmHg;(142.6? 13.8)bpm vs.(124.6?12.5)bpm;P0.05].For the intubation condition,the satisfactory rate in inhaled group was higher than that of intravenous group,P

Objective To investigate the differences in the structures,numbers and phenotypes of the dendritic cells (DCs)at different stages in colorectal cancer (CRC)tissue,and to explore the effect of DCs in the occurrence and development of CRC.Methods 20 cases of CRC tissue were chosen as CRC group,other 20 cases of corresponding para-cancer tissue were chosen as control group.Immunohistochemical staining was used to detect the distribution, the structures,the number of positive cells at different stages;Western blotting was used to detect the expressions of CD1a and CD83.Results The results of immunohistochemical staining showed that DCs scattered among glands or vessels in both groups;the CD1a+imature DCs (imDCs)were round and oval,dendritic processes were faintly visible in both groups,and the number of CD1a+imDCs in experimental group was significantly higher than that in control group (P<0.01).The CD83+ mature DCs (mDCs)were irregular,branched in both groups,and the number of CD83+mDCs in CRC group was significantly lower than that in control group (P<0.01).The results of Western blotting method showed that the expression of CD1a protein was higher and the expression of CD83 protein was lower in CRC group than control group (P<0.01).Conclusion CD1a+imDCs may play important roles in promoting the occurrence and development of CRC;CD83+mDCs may inhibit the occurrence and development of CRC.

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 rapid genotyping method of for methicillin-resistant Staphylococcus aureus(MRSA) based on polymerase chain reaction(PCR)-high resolution melting (HRM ) curve analysis and staphylococcal protein A (SPA ) classifica-tion .Methods 71 strains of MRSA clinically isolated were collected as test strains .Gene sequencing and HRM curve analysis were employed to conduct SPA gene typing .Results According to gene sequencing method ,SPA gene of 71 strains of MRSA was divided into four types ,namely t570 ,t030 ,t002 and t588 .The most predominant type was t570 (74 .65% ) ,followed by t030 and t002(both 7 cases) .The result of SPA gene typing by HRM analysis were basically consistent with that by gene sequencing .Con-clusion PCR-HRM analysis is expected to become a fast ,efficient genotyping for MRSA SPA gene ,providing the basis for hospital infection control .

Objective To investigate celecoxib self-microemulsifying drug delivery system (CXB-SMEDDS) that was developed to increase the dissolution rate and oral bioavailability of celecoxib.Methods The formulation of CXB-SMEDDS was optimized by pseudo-ternary phase diagrams analysis.The appearance, morphology, particle size distribution and in vitro drug release behavior of CXB-SMEDDS were investigated after diluted by water.The bioavailability of CXB-SMEDDS was determined by oral administration to rats compared with CXB suspension.Results An optimized formulation was selected: Medium chain triglycerides as oil phase, Tween 20 as surfactant, Transcutol HP as cosurfactant.The ratio of oil phase, surfactant and cosurfactant was 2∶9∶9.Upon mixing with water, CXB-SMEDDS formed a clear and transparent microemulsion solution with homogeneous small spherical under transmission electron microscopy.For particle size of CXB-SMEDDS was found to be (57.6±14.2) nm.The in vitro dissolution test indicated a significant improvement in release characteristics of CXB.The AUC of CXB-SMEDDS and CXB suspension were (5.54±0.94) and (3.32±0.59) mg·L-1·h, respectively.The relative bioavailability was 166.9%.Conclusion The SMEDDS can significantly increase celecoxib dissolution in vitro and bioavailability in vivo.

Objective:To analyze the expression of E-cadherin in the epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in neuroblastoma.Methods:TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), was applied to SK-N-SH cells in vitro compared with the blank control group.EMT-related genes mRNA and protein expression were detected by carrying out real-time PCR assays and Western blot.A scratch test and migration assay were performed to verify the alteration of SK-N-SH cell migration capacity.Data collected from 18 cases of neuroblastoma patients were selected from the Department of Hematology Oncology, Shanghai Children′s Hospital from January 2008 to December 2012.The expression of E-cadherin in the tumor tissue of the neuroblastoma patients after operation was detected by immunohistochemistry.The clinical features and survival prognosis of these patients were analyzed. Results:Compared with the control group, after SK-N-SH cells were treated with TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), real-time PCR assays and Western blot revealed that the mRNA(0.603±0.081, 0.606±0.008, 0.716±0.166 vs.1.000) and protein expression levels(0.855±0.026, 0.600±0.017, 0.495±0.011 vs.1.000) of E-cadherin were significantly decreased ( F=8.144, P=0.040; F=74.810, P<0.001), while the mRNA(2.132±0.167, 3.494±0.017, 4.184±0.021 vs.1.000) and protein expression levels (1.175±0.053, 1.189±0.058, 1.225±0.106 vs.1.000)of α - smooth muscle actin were significantly increased ( F=547.300, P<0.001; F=68.810, P=0.007), suggesting that EMT changes occur in cells.Scratch test and Transwell migration assay revealed that the number of migrating cells increased obvious with the treatment of TGF-β1 (5 μg/L) ( t=16.070, P=0.040). The 10-year overall survival(OS) rates of neuroblastoma patients with E-cadherin strong positive expression, positive expression, weak positive expression and negative expression in the pathology were (77.78±13.86)%, (75.00±21.66)%, (25.00±21.65)% and 0, respectively ( F=8.160, P=0.040). Conclusions:TGF-β1 can induce the EMT in SK-N-SH cells and increase cell migration.The decrease expression of E-cadherin in neuroblastoma patients is closely associated with clinical progress and recurrence or metastasis of the disease.

Objective To analyze the clinical data of children with Langerhans cell histiocytosis (LCH),to discuss the therapeutic effect,and to analyze the factors related to prognosis.Methods A total of 45 children diagnosed as LCH were divided into group A (18 cases with bone lesion only),group B(6 cases with soft tissue lesion),and group C (21 cases with viscera lesion) according to Shanghai Children's Hospital-LCH-2007 scheme [SCH-LCH-2007 (modified DAL-HX83/90) scheme].(1) Initial treatment:group A was treated with Prednisone (Pred) + Vincristine (VCR) for 28 weeks,and group B was treated with Pred + VCR + Etoposide (VP16) + Mercaptopurine (6MP) for 43 weeks,and group C was treated with Pred + VCR + VP16 + Methotrexate (MTX) +6MP for 52 weeks.(2) Re-treatment scheme after relapse included:①upgrading treatment,group A to group B,group B to group C.②Individual treatment for group C included VP16 modification,and maintained Thymosin and/or Ciclosporin etc.Results The total survival rate was 93.3％ (42/45 cases) and recurrence rate was 26.7％ (12/45 cases).Children in group A and B were all effective,while 2 patients in group C died,and 1 case missed follow-up.Multi-factor analysis showed that the factors like age,sex,group,skeleton,soft tissue,erythra,lymph gland,lung,mouth,ears,hypophysis pituitary had no statistical significance,but liver,spleen and blood involvement had statistical significance in disease relapse:liver (P=0.007 1),spleen (P=0.016 9),and blood (P=0.011 1).Conclusion LCH can affect several organs of children and relapse,and modified DAL-HX83/90 scheme is very effective.The liver,spleen and hematopoiesis system involvement is correlates with the relapse.

Safety hospital centralized monitoring is one of the important methods for evaluating the medication safety of TCM injections in clinic.It is critical to warrant research quality by reasonable statistical analysis.However,universally accepted standards and guidelines for the statistical analysis of hospital centralized monitoring of TCM injections have not been issued so far,which would probably reduce the research quality of safety hospital centralized monitoring of TCM injections since the unreasonable use of statistical analysis methods.Combined with previous practical experience and understanding,we put forward the problems of statistical analysis of hospital centralized monitoring of TCM injections with the provision of some advice in this paper based on the analysis of the preceding studies,laying a foundation for the same kind of researches.