Objective This study is undertaken to evaluate the changes of serum cytokine levels in different stages of collagen induced arthritis(CIA)rats,to search for the specific proteins related with rheuma- toid arthritis(RA)pathogenesis and inflammation,and to explore the mechanism of RA pathogenesis.Methods Rat cytokine antibody array coated with 19 specific cytokine antibodies was used to examine serum samples at peak and late stage of CIA rats,and were compared to normal cytokine levels.At the same time,ELISA assay for serum TNF-?production was used to verify the array results.Results Among the target cytokines,10 up- regulating cytokines were kept in high expression in different phases of disease,while 1 showed significant change only at the peak of disease.There was no downregnlating cytokines in the results.Serum TNF-?assay results were consistent to the array results.Conclusion Cytokines show different expression in CIA at differ- ent stages,and specific cytokines can be used as the candidates to further study of the RA pathogenesis.This study also provides molecular makers for early diagnosis.

Objective To develop paclitaxel-loaded polymeric micelles from poly (ε-caprolactone)-poly (ethylene glycol)-poly(ε-caprolactone) (PCL-PEG-PCL),and to evaluate in vitro cytotoxicity as well as in vivo antitumor activity against EMT-6 tumor breast cell.Methods Paclitaxel-loaded polymeric micelles were prepared by thin-film hydration and ultrasonic method.The physical status of paclitaxel inside the polymeric micelles was investigated by differential scanning calorimetry (DSC).In vitro cytotoxicity of paclitaxel-loaded polymeric micelles against EMT-6 cell line was assessed by MTT assay.In vivo anticancer activity was evaluated against EMT-6 tumorbearing mice,with commercially available Taxol injection as control.Results Paclitaxel-loaded polymeric micelles exhibited homogeneous spherical shapes with apparent core-shell morphology.The average diameter of paclitaxelloaded polymeric micelles was 93 nm.DSC study indicated that paclitaxel was in solid amorphous state after being encapsulated in the polymeric micelles.In vitro cytotoxicity demonstrated that the cytotoxic effect of paclitaxelloaded polymeric micelles was lower than that of Taxol injection at the same paclitaxel content.Paclitaxel-loaded polymeric micelles showed greater tumor growth-inhibition effect in vivo on EMT-6 breast tumor in comparison with that of Taxol injection,with tumor growth inhibition of 85.79％ and 63.37％,respectively (P＜0.05).Conculsions The prepared paclitaxel-loaded polymeric micelles showed high anti-tumoral efficacy and low toxicity,and might have the potential to be developed as an effective anticancer drug-delivery system for cancer chemotherapy.

Humans ; Intestines ; Stomach Neoplasms/complications*

Histocytochemistry ; methods ; Humans ; Pathology, Clinical ; methods ; Specimen Handling ; methods ; Staining and Labeling ; methods

Objective To investigate the clinical features of and GJB2 gene mutations in a Chinese Han patient with keratitis-ichthyosis-deafness syndrome (KID syndrome),in hope to offer evidence for the clinical and genetic diagnosis of KID syndrome.Methods Clinical data were collected from a patient with KID syndrome.DNA was extracted from peripheral blood of the patient and his two family members (mother and brother).PCR was performed to amplify the exon 2 and its flanking splicing sites of GJB2 gene followed by bidirectional direct DNA sequencing. Results The patient presented with the typical triad of vascularizing keratitis,ichthyosis and congenital deafness.A G148A mutation in the exon 2 of GJB2 gene,resulting in the substitution of aspartic acid by asparagine at position 50 of the junction protein connexin 26 (Cx26),was identified in the patient,but not in either of his family members.Conclusion The G148A mutation in GJB2 gene may be responsible for the clinical phenotype of KID syndrome in this Chinese patient.

ObjectiveTo isolate and identify endothelial progenitor cells (EPCs) from human umbilical cord,and to study the cell proliferation and gene transfection of green fluorescent protein plasmid in vitro.MethodsEPCs were isolated from human umbilical cord in enzyme digestion method.The biological characteristics of EPCs were identified by flow cytometry and laser confocal microscope.The enhanced green fluorescent protein (EGFP) gene transfection mediated by EPCs was investigated using Lipofectamine 2000 as transfection reagent.ResultsEndothelial progenitor cells isolated from umbilical cord formed typical endothelial cell colony 9 days later.These cellsdisplayed an improved positive expression of CD133 and kinase insert domain receptor (KDR).The endotheliallineage characteristics of expanded cells were confirmed by fluorescein isothiocyanate (FITC)-UEA-1 binding and DiI-ac-LDL uptake assay with the aid of laser confocal microscope.The transfection results demonstrated high expression of EGFP taking EPCs as host cell.ConclusionEndothelial progenitor cells isolated from umbilical cord can be propagated and induced to differentiate into endothelial cells in the appropriate culture conditions.EPCs demonstrated to be an ideal carrier for gene and cell therapy.

The SRY gene from buffalo (Bubalus bubalis) genome was amplified by the polymerase chain reaction ( PCR) with primers based on the sequence of Hostein SRY gene. The amplified fragment was 2005 bp include 5UTR ( 1 - 504bp) and 3'UTR(1196 - 2005bp). And the amplified fragment was cloned and sequenced. Sequence analysis showed that the coding region of SRY gene (505 - 1195bp) from buffalo was highly homologous with those of other bovine counterpart genes (96% homology) , especially in the HMG box region (99%homology). It was found that there were only signal on male buffalo genome on Southern blot,which indicate SRY gene are highly conservative on evolves.

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Objective To evaluate the correlation between cerebral blood volume and permeability surface by using muhislice CT perfusion imaging with glioma grade.Methods Ninteen patients with gliomas underwent conventional MR and multislice CT perfusion imaging preoperatively.These patients were divided into low grade and high grade groups which were correspond to WHO Ⅱ grade gliomas and WHO Ⅲ or Ⅳ grade gliomas respectively.CT data were transferred to on-line working station and processed to obtain time-signal curves,color perfusion maps and calculated perfusion parameters,including cerebral blood volume(CBV),cerebral blood flow(CBF),mean transit time(MTT)and permeability surfaces (PS)in tumoral parenchyma.Kruskal-Wallis test and correlation of CBV and PS was assessed by using SPSS 11.0 software.Results The median of CBV and PS in low-grade and high-grade glioma were 2.7, 6.5 ml/100 g;0.389,12.810 ml?100 g~(-1)?min~(-1),respectively,corresponding t value were 12.907, 13.500 with P

Objective To study the effect of once-weekly injection of recombinant human parathyroid hormone rhPTH(1-34) on the healing of bone fracture in rats.Methods Fifty male 3-month old SD rats were used in this study to produce unilateral tibial fracture and received internal fixation with a Kirschner needle.Based on the dose and frequency of rhPTH (1-34) injection, the rats were randomly divided into five groups (n=10 each) as follows: subcutaneous injec-tions of saline, and rhPTH in a dose of 10, 20, 10, and 20 μg/kg/d.After 4 weeks of treatment, the rats were euthana-tized and the fractured tibia were assessed by X-ray, dual energy X-ray absorptiometry ( DXA) ,micro-computed tomography ( microCT) and three-point bending test.Results The fracture healing in the 20μg/kg/w group was better than the saline group.The fracture healing in the 20μg/kg/w group was as well as the 10 μg/kg/dgroup.The BMD of 20μg/kg/w group was 26.2%higher than the saline group.The mineralized callus volume in the 20μg/kg/w group was 51.4%higher than the saline group.The total callus volume in the 20μg/kg/w group was 21.6%higher than the saline group.The ultimate load of the 20 μg/kg/w group was 29.3% higher than the saline group.There was no significant difference between the 20 μg/kg/w group and 10 μg/kg/d group in BMD, bone micro-architecture, and biomechanical strength ( P >0.05 ) . Conclusions Once weekly injection of rhPTH (1-34)can promote the bone fracture healing.