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*

The formulation for sustained release tablet of Epinedium component was selected and the evaluation equation of in vitro release was established. The liquidity of component was improved with the help of colloidal silica aided by spray drying, which would be the main drug in the sustained release tablets. Dissolution was selected as an evaluation index to investigate skeletal material type, fillers, impact porogen, lubricants and other materials on the quality of sustained release tablet. The sustained release tablets were prepared by dry compression. Formulation of sustained release preparations was main drug 35%, HPMC K(4M) 20% and HPMC K(15M) 10% as skeleton material, MCC 31% as filler, PEG6000 2% as porogen and magnesium stearate 2% as lubricant. The sustained release tablets released up to 80% in 8 h. The zero order equation, primary equation and Higuchi equation could simulate the release characteristics of sustained release tablets in vitro, the correlation coefficients r were larger than 0.96. The primary equation was most similar in vitro release characteristics and its correlation coefficient r was 0.9950. The preparation method is simple and the results of formulation selection are reliable. It can be used to guide the production of Epimedium component sustained release preparations.
Chemistry, Pharmaceutical ; methods ; Delayed-Action Preparations ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Epimedium ; chemistry ; Kinetics ; Tablets ; chemistry

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.

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.

Objective To investigate the changes of vital signs and the damage of important organs in dog progressive circulatory failures induced by soman.Method Seven male dogs,weighing(12～15)kg,were injected intramuscularly 1/3 LD sornan(1 LD=10μg/kg)per ten minutes.The moan blood pressure decreased to (40～45)mmHg was defined as circulatory failure.The changes of heart rate,blood pressure.and hemodynamic parameters were evaluated by an eight-channel direct-witing oscillograph,blood gas,pH value,electrolyte,and the damage of important organs were observed before and after sornan injection.Statistical analysis of the data was performed using the self control t test with the SAS 6.12 Software Program.Results In anesthetized dogs intoxicated with sornan,the circulatory failure was characterized by the significant decreases in blood pressure,heart rate and hemodymrnic parameters(P<0.05).Partial pressure of oxygen was less than 60 mmHg,saturation of oxygen Was less than 90% and partial pressure of carbon dioxide was greater than 50 mmHg in arterial blood of the dog model.These results showed mix respiratory failure occurred during intermittent positive pressure.Significant metabolic acidosis was induced by soman[pH(7.345±0.064)vs.(6.956±0.022),P<0.01].The concentralion of sodium ion and chloride ion in blood were changed gently.The concentrations of GTP,GOT,Cr,BUN,CK-MB and LDH were increased significantly(P<0.05),which showed multiple important organs including liver,kidney and heart were damaged by sornan.Conclusions The severe progressive circulatory failure induced by cholinesterase inhibitor sornan leads to the darnage of vital signs and important organs significantly.

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

OBJECTIVETo investigate the clinical application of free superficial iliac circumflex artery skin flaps, as well as the management of donor site defects.

METHODS17 free superficial iliac circumflex artery skin flaps were applied for the traumatic defects or deformities on face, neck, foot, hand, ankle and lower leg, respectively. The donor site defects were closed directly or covered by paraumbilical island flaps.

RESULTSThe 17 flap size ranged from 5 cm x 3 cm to 19 cm x 14 cm. 16 flaps survived completely except 1 flap with partial necrosis, which was closed by free skin graft. The donor site defects were closed directly in 10 cases, and covered by paraumbilical island flaps in 7 flaps without no flap necrosis. The abdomen had a good appearance.

CONCLUSIONSGood appearance can be achieved with free superficial iliac circumflex artery skin flaps for the defects on face, neck, foot, hand, ankle and lower leg. Paraumbilical island flap can be used for the donor site defects.

Arteries ; Foot ; Free Tissue Flaps ; blood supply ; transplantation ; Humans ; Reconstructive Surgical Procedures ; Skin ; Skin Transplantation ; Transplant Donor Site ; surgery ; Wounds and Injuries ; surgery

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