Objective To analyze the relationship between dynamic change of GPI-80 and disease severity and prognosis of childhood anaphylactoid purpura. Methods Patients were collected and divided into three groups according to their clinical features: purpura group (purpura only), mixed group (purpura + arthritis + gastrointestinal bleeding) and nephritis group. There were 20 patients in each group. GPI-80 expression on the neutrophils was detected by flow cytometry during acute and regressive phases of the disease. GPI-80 expression was compared among different groups and different phases. Renal biopsies were performed in 20 nephritis patients. Results GPI-80 expression was significantly increased in all patient groups compared with that in the normal control (P 0.05). No significant difference of GPI-80 expression was found among 20 nephritis patients with different pathological patterns. Forty-two patients (10 in purpura group, 15 in mixed group, and 17 in nephritis groups) were followed up and GPI-80 expression was detected at the time of discharge and 2 weeks after discharge, the results showed that GPI-80 expression was decreased from 93.26% (?7.89%) at acute phase to 91.37% (?6.9%) at regressive phase with an average interval of 13.5 days. Most of them (35/42) further decreased to 38.44% (?7.8%) in 2 weeks after discharge. GPI-80 expression remained high in 7 patients for 2 weeks after discharge and relapsed in 5 patients within 1 month after discharge. Conclusions High GPI-80 expression is related to the severity of the disease. The decrease of GPI-80 takes place later than the improvement of clinical symptoms. Children with persistently high GPI-80 expression are likely to relapse. It seems that there is no correlation between GPI-80 expression and different pathological patterns of nephritis.

Double cannulation model of conscious rat allowing simultaneous collection of mesenteric lymph and jugular venous blood was established to investigate the intestinal lymphatic transport of breviscapine orally administered in rat. The concentrations of breviscapine in plasma and lymph were determined by HPLC. The pharmacokinetics of breviscapine after oral and intravenous administration was evaluated in the conscious rat model. It was observed that scutellarin distributed from blood circulation to lymphatic system after intravenous injection. The cumulative lymphatic transport amount within 12 h was (2.78 +/- 0.25) microg, equivalent to 0.0792% of intravenous dose. After oral administration of scutellarin to double-cannulation rats, the cumulative lymphatic transport amount within 12 h was (0.92 +/- 0.08) microg, equal to 0.0083% of oral dose. The absolute bioavailability of breviscapine orally administered to double-cannulation rats was 4.91%, indicating that scutellarin was mainly absorbed into the bloodstream through the portal vein. Lymphatic transport of scutellarin appears to reflect high affinity for the lymph lipoproteins to chylomicron. This study provided a biopharmaceutics basis for developing oral lipid delivery system for the promotion of intestinal lymphatic transport to improve oral bioavailability of breviscapine.

s were 0.573±0.016, 0.707±0.008 and 0.711±0.013). Conclusions CXCL10 can express stablely in MCF-7 cell lines, which resulted in down-regulation of expression of VEGF and STAT3 gene. CXCL10 played an important role in anti-tumor effect.

AIM: To evaluate the implication of CXCL10-loop3-EGF fusion protein for the activities of targeting tumor and anti-angiopoiesis. METHODS: RT-PCR was preformed to amplify CXCL10 coding sequence from PBMC activated by IFN-γ. CXCL10-loop3-EGF fusion gene, which was conducted by Over-Lap Extention PCR, was hinged up with plasmid pTG19-T, transfected to E. coli DH5α and processed positive colony selection. After ligated with plasmid pET32a(+), recombinant CXCL10-loop3-EGF fusion gene was then transfected to E. coli Origami B (DE3) and induced to express its coding fusion protein his-CXCL10-loop3-EGF. The recombinant fusion protein CXCL10-loop3 -EGF was purified by His-bind affinity chromatograph, enterokinase cleavage, ultrafiltration and dislysis. The transwell chemotatic test and HUVEC angiopoiesis inhibition test were performed to determine the anti-tumor responses and anti-angiopoiesis activity of CXCL10-loop3-EGF fusion protein. RESULTS: CXCL10-loop3-EGF fusion protein was successfully constructed and confirmed by SDS-PAGE analysis and Western blotting. Significant PBMC chematatic activity and HUVEC anti-angiopoiesis activity were observed. CONCLUSION: CXCL10-loop3-EGF fusion protein, which has perfect anti-tumor activity, is successfully constructed.

Objective To investigate the risk factors on rectovaginal fistula after resection in rectal canc-er and clinical strategy. Methods Our retrospective study included 1123 patients of recter cancer who un-derwent anterior resection with TME technique. Results 3.03% patients(34/1123) developed reetovaginal fistula. Rectovaginal fistulas were raleted with menopausal, location tumor in rectal wall and distance between tumor and anal verge, anastomotic technique, while not with age, T stage, preoperative radiotherapy, diversion stoma construction. Among 34 patients 12 were cured conservatively, fistula repair and colon stoma were performed for the other patients. Conclusions Menopausal, location tumor in rectal wall, distance between tumor and anal verge, anastomotic technique were risk factors for rectovaginal fistula after resection in rectal cancer. Correct choice of surgical procedures, suitable operative timing, enough preoperative preparation are important.

Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

Background and purpose:We investigated whether lfuorine-18 lfuorodeoxyglucose (18F-FDG) maximal standard uptake value (SUVmax) of the primary tumor (SUV-T), SUVmax of the regional lymph nodes (SUV-N) or the overall loco-regional lesion SUVmax (SUV-TOTAL) was related to survival of patients with stage Ⅲ non-small cell lung cancer (NSCLC) who received Cetuximab and combined definitive chemoradiotherpay. Methods:From September 2009 to July 2012, seventeen patients with unresectable stageⅢNSCLC receiving cetuximab with cisplatin/vinorelbine (NP) followed by concomitant NP and intensity-modulated radiotherapy (IMRT) at the Fudan University Shanghai Cancer Center were enrolled onto a prospectively study. All patients received positron emission tomography/computerized tomography (PET/CT) scans within 2 weeks before enrolment. Univariate analysis were used to assess the correlation between SUV-T, SUV-N, SUV-TOTAL, gender, age, histology, tumour-node-metastasis (TNM) stage, performance status (PS) as well as smoking status and survival. The factors which showed statistical signiifcance entered into multivariate Cox-regression model. Survival functions of different populations were estimated by Kaplan-Meier method and compared by Log-rank test. Results:In the univariate analysis, SUV-T, SUV-N, SUV-TOTAL, PS and smoking status were prognostic factors. The best cut-off values for SUV-T, SUV-N and SUV-TOTAL were 11, 11 and 20, respectively. Multivariate analysis revealed that SUV-TOTAL (P=0.012), SUV-T (P=0.025), and SUV-N (P=0.033) were independent predictors of survival with hazard ratio (HR) of 14.7, 11.2, and 6.2, respectively. Conclusion:Local, regional and locoregional maximal SUVs deifned by 18F-FDG PET-CT scanning may have a strong correlation with survival in this patients setting, which merits further study.

The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

Objective: To explore the anti-proliferation and apoptosis-inducing effects of decitabine combined with paclitaxel on paclitaxel-resistant MCF-7 cells. Methods:Cell counting kit-8 (CCK-8) assays were used to determine the proliferation inhibition of drugs at different concentrations in 24, 48 and 72h. The group was treated with decitabine, paclitaxel and the combination of the two drugs, respectively. The cell apoptosis rate was determined by flow cytometry after the treatment with the drugs at different concentra-tions in 48h. Results:The results of CCK-8 showed the combination group significantly inhibited the cell proliferation when compared with the single drug use group(P<0. 05), and the anti-proliferation value was in a dose-dependent manner in all groups. The apopto-sis values after the treatment with decitabine, paclitaxel and the combination in 48h was(20. 4 ± 1. 98)%,(21. 8 ± 3. 34)% and(70. 8 ± 8. 23)%,respectively. Conclusion:The proliferation inhibition and apoptosis induction are both increased significantly in the com-bination group. Synergistic effect of decitabine and paclitaxel is found in multidrug resistant breast cancer cell line MCF-7 in vitro.

Objective To investigate the influence of IL-8 on the tight junction of vascular endothelial cells.Methods Immunofluorescence was used to observe the modality and the distribution of three tight junction proteins (occludin,claudin-5 and ZO-1) of the EA.hy926 cells treated with IL-8 under different concentrations and different times.RT-PCR was used to measure the mRNA expression of these three proteins.Results The results demonstrated that IL-8 could change the distribution of occludin,claudin-5 and ZO-1 in EA.hy926 cells,and the mRNA expression of occludin,claudin-5 and ZO-1 decreased with the increase of IL-8 concentration and treated time.Conclusion The effects of IL-8 on the distribution and the expression of occludin,claudin-5 and ZO-1 are dose and time-dependent.