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 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 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 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 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 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 investigate the clinical results and related key points of surgical treatment for Hawkins Ⅲ talus neck fractures.Methods From March 2005 to March 2010,26 patients with Hawkins Ⅲ talus neck fracture were treated and 21 of them were followed,including 13 males and 8 females,with an average age of 37.6 years.The fractures occurred on the left side in 11 patients and on the right side in 10patients.The mechanism of injury included high falling injury in 13 patients,traffic accident injury in 7 patients and rolling down injury in 1 patient.Five cases were closed fractures and 6 cases were open fractures.A bilateral approach,the medial and lateral approaches,was used to perform the operation with cannulated screw fixation in emergency.After external fixation in functional position for 6 weeks without loading,ankle joints were allowed to take exercise with hinge brace and to bear partially basing on plain radiograph 12-16weeks later.The weight-bearing should be adjusted with follow-up.Functional results were assessed according to AOFAS (American Orthupaedic Foot and Ankle Society,AOFAS)score.Results The average duration of follow-up was 36.6 months (range,6-60 months).All fractures gained union and the average union time was 4.5 months.The average AOFAS score was 78.6.There were 4 cases in excellent results,10 in good,5 in fair and 2 in poor.The overall excellent and good rate was 67.8％.Traumatic arthritis occurred in 13 cases and avascular necrosis in 5 cases.Conclusion The effect of surgical treatment for Hawkins Ⅲtalus neck fracture via a bilateral approach is satisfactory.

Objective To evaluate the efficacy of different treatment regimens for children with acute promye-locytic leukemia (APL)with positive PML -RARa fusion gene.Methods Thirty -two newly diagnosed APL patients were included in this study,treated either with all -trans -retinoic acid (ATRA)and chemotherapy (CT)(group A) or with ATRA and arsenic trioxide (ATO)(group B).Clinical situation and clinical efficacy were analyzed in patients in different groups.They were also separated into low risk group,intermediate risk group and high risk group according to different risk criteria.Clinical characteristics,complete remission,long -time survival and urine arsenic concentra-tion were analyzed and compared.Results (1 )Fourteen of 1 5 patients (93.3%)in group A achieved hematological complete remission (HCR)with a median time of 38 days (28 -63 days).Sixteen of 1 7 patients (94.1 %)in group B achieved HCR with a median time of 29 days (1 0 -42 days),which was significantly shorter than group A,and there was a significant difference between 2 groups(t =3.53,P =0.002).(2)The 5 -year event -free survival (EFS)of group A and group B was (60.0 ±1 2.6)% and (81 .9 ±9.5)%,respectively;the 5 -year EFS of group B was almost 20% higher than group A;while there was no significant difference between the 2 groups(χ2 =1 .1 5,P =0.28).The 5 -year overall survival (OS)of group A and group B was (72.2 ±1 1 .9)% and (94.1 ±5.7)%,respectively,the 5 -year OS of group B was almost 20% higher than group A;while there was no significant difference between the 2 groups(χ2 =2.88,P =0.1 6).(3)The 5 -year EFS of low plus intermediate group and high risk group patients was (74.0 ±1 0.1 )% and (64.8 ±1 4.3)%,the 5 -year EFS of low plus intermediate group was almost 1 0% higher than high risk group,but there was no significant difference between the 2 groups(χ2 =0.1 4,P =0.71 ).The 5 -year OS of low plus intermediate group and high risk group patients was (84.7 ±8.1 )% and (71 .3 ±1 4.1 )%,the 5 -year OS of low plus intermediate group was almost 1 0% higher than high risk group,while there was no significant difference be-tween the 2 groups(χ2 =0.36,P =0.55).(4)ATO related side effects were mild,including abnormal liver tests and e-lectrocardiogram,but were invertible after supportive therapy.At the end of each chemotherapy course,the urine arsenic concentration remained low and no chronic arsenic toxicity or second malignancies were found during the follow -up period.Conclusions The ATRA plus ATO regimen is a promising and better treatment for childhood APL with positive PML -RARa fusion gene compared with conventional chemotherapy.It was necessary to take risk stratification in APL patients.