The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM's proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM's proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.
Absorbable Implants ; Basement Membrane ; Biocompatible Materials ; chemistry ; Epithelium ; Esophagus ; physiology ; Fibroins ; chemistry ; Humans ; Nanostructures ; chemistry ; Polyesters ; chemistry ; Regeneration ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Antigens, CD19 ; analysis ; Asthma ; blood ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Immunoglobulin E ; blood ; Lymphocytes ; immunology ; Male ; Receptors, IgE ; analysis

Objective To investigate the methods of isolation,culture,identification and labeling of mesenchymal stem cells(MSCs) in vitro and lay a foundation for further study on intervention of MSCs on immunologic rejection of organ transplantation. Methods MSCs were isolated and cultivated by adherent methods . The expressions of CD90 and CD45 of cells were analyzed by using flow cytometry in order to identify MSCs.The third generation of MSCs were labeled by DAPI,the labeling efficiency was detected.Results Primary cultured MSCs adhered to plastic surface within 48 h and reached 90% confluence within 7-10 d .Flow cytometry showed that the positive rates of CD90 and CD45 of MSCs at third generation were 99.8% and 6.8%. MSCs expressed CD90 but no CD45.All of the MSCs after labeling by DAPI showed blue fluorescence by immunofluoroscope. DAPI labeling was sensitive and highly efficient to MSCs.Conclusion Adherent method is simple and easy to isolate and cultivate MSCs and it can serve as a routine method.DAPI labeling can be used as a efficient method to label MSCs.

Objective To design and prepare an RNA interfering vector for effectively inhibiting the cellular expression of zinc finger protein A20 and observe the effect of A20 gene silence on cellular inflammatory response. Methods Specific RNA interfering oligonucleotide fragments (ASRF) were designed and synthesized artificially and the A20 RNA interfering vector pSUPER-EGFP-A20 siRNA constructed. Human monocyte cell line THP1 was used to infect the pSUPER-EGFP-A20 siRNA by means of genetic transfection technique; then, silence rate of cellular A20 was analyzed by real-time polymerase chain reaction (PCR). In the meantime, the activity of nuclear transcription factor nuclear factor-κB (NF-κB) and the level of tumor necrosis factor-α (TNF-α) in culture supernatant were measured by ELISA. Results Of two specific inhibitory oligonueleotide fragments of A20, the fragment M59465-385R/F had a higher inhibition to A20 expression, with rate of A20 gene silence of 83.86%. Preliminary application showed that after A20 gene silence, the activity of NF-κB was increased by 78.13% and the level of TNF-α in cell culture supernatant was increased by 49.30%. Conclusions Vector of A20gene silence with a high efficiency is obtained successfully. Preliminary application indicates that the expression of A20 can down-regulate the degree of cellular inflammatory responses.

Objective To assess the nutritional risk of patients with inflammatory bowel disease (IBD).Methods The nutritional status of 112 IBD patients from PUMC Hospital were evaluated by Nutritional Risk Screening 2002 (NRS 2002).Using Chinese standard,a body mass index (BMI) that was lowered than 18.5 kg/m~2 according to clinical material was regarded as malnutrition,and the score was recorded as 3.The se-verity of ulcerative colitis (UC) was evaluated using True-Love criteria as mild,moderate,and severe.Crohn's disease (CD) was evaluated using Harvey-Bradshaw Index as in remission stage (≤4),moderately active stage (4-8),and severely actively stage (≥9).Results All these 112 patients,including 70 UC cases and 42 CD cases,were evaluated by NRS2002.Forty-five patients (40.2%) were judged as at the risk of malnutrition.The incidence of malnutrition was significantly higher in CD patients than in UC patients (52.4% vs.32.9%,P<0.05).It was also significantly correlated with the severity of disease.In patients with severe IBD,the rate of mal-nutrition evaluated by BMI according to clinical material was significantly lower than the rate of nutritional risk.Con-clusions The risk of malnutrition is high in IBD patients.NRS 2002 can be used for nutritional risk screening.

Objective To construct a novel tissue?engineered bone matrix gelatin (BMG)/poly[butylene succinate?co?tere?phthalate] (PBST) biphasic scaffold for annulus fibrosus. Methods The PBST spinning fibers were prepared by electrospinning and the porosity and water absorption rate were tested. Rabbit annulus fibrosus cells were isolated, cultured and identified through SafraninOstaining, and collagenⅡimmunohistochemical staining in vitro. And then annulus fibrosus cells were implanted on the PBST fiber, whose growth situation was observed by scanning electron microscope (SEM). Then the BMG/PBST biphasic scaf?fold was constructed by BMG as the outer annular fibrosus and PBST fiber as the inner annular fibrosus. The annulus fibrosus cells were implanted on the biphasic scafflod and cultured for 3, 7 and 21 days in vitro. The biomechanical and biological property was observed at the predetermined time point. Results The porosity of the fiber was 61.83%±7.33%and its water absorption rate was 297.34%± 57.13%. The identified result of annulus fibrosus cells were positive, suggesting that the cells have still kept their annulus fibrosus cells characteristics. The cells growth could be observed through SEM at 3rd and 7th day after implanted on the fi?bers. After cultured on the BMG/PBST scaffold, HE staining proved that the cells could ingress into the inner of fiber with time. SafraninOstaining and collagenⅡimmunohistochemical staining proved that the cells can secreted abundant proteoglycan and collagenⅡ, the special annulus fibrosus cell extracellular matrix. Compared with the BMG/PBST scaffold without cells, the elastic modulus of biphasic scaffold was increased from 14.83±1.02 MPa to 17.56±1.47 MPa after cultured with cells for 21 days in vitro. Conclusion The novel tissue?engineered biphasic scaffold for annulus fibrosus constructed with BMG and PBST fiber spinning has good cytocompatibility and biomechanical characteristics, which provide a basis for the complete tissue engineered interverte?bral disc.

Objective To understand the comments for the essential drug system by doctors at township health centers in Nantong's six counties and to discuss the effect of implementing the essential drug system to different ownership systems.Methods The survey was made with stratified clusters random sampling method to select township health centers.Questionnaires were filled out by doctors,and leaders of the health bureau and township health centers were interviewed.Results After the implementation of the essential drug system,94.6％of the doctors found that the number of laboratory examinations increased in Rugao,Rudong and Haian.23.2％of the doctors in Tongzhou,Qidong and Haimen found that enthusiasm for work increased.Conclusion The compensation mechanism should be perfected and the financial subsidies should be paid in time.Supervision and management should be strengthened and the grassroots need for medicine be ensured.The skills training should be enhanced and the medical service ability be improved.

BACKGROUND:Polyurethane has good mechanical and physical characteristics and is extensively used in
clinical and experimental studies, but its hydrophobicity and histocompatibility are not ideal, which limits its use in tissue engineering as a biomaterial scaffold to some extents.
OBJECTIVE:To observe the hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin and its compatibility with human hypopharyngeal cel s.
METHODS:The changes in hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin were detected by contact angle measurements. Human hypopharyngeal fibroblasts were cultured in vitro on
polyurethane membrane, silk fibroin-polyurethane membrane, glutin-polyurethane membrane and tissue culture plate. Cel compatibility was compared using cytometry and cel morphology obsevation.
RESULTS AND CONCLUSION:Hydrophilicity of silk fibroin-or glutin-polyurethane membranes significantly increased (P<0.01). The hydrophilicity of silk fibroin-polyurethane membrane was higher than that of
glutin-polyurethane membrane (P<0.01). The number of cel s on the tissue culture plate was the most. The number of human hypopharyngeal fibroblasts on the silk fibroin-or glutin-polyurethane membrane was higher
than that on the polyurethane membrane, especial y on the silk fibroin-polyurethane membrane. These suggested that hydrophilicity and cel compatibility of silk fibroin-or glutin-polyurethane membrane were elevated.

Objective To supply evidence for quality control of Herba Ephedra by determining content limit in different batches of medicinal material of Herba Ephedra.Methods A RP-HPLC method was used for the determination of ephedrine and pseudo-ephedrine. The chromatographic conditions were: Kromasil RP-C18(250?4.6 mm,5 ?m) column, a mixture of 0.3 %phosphoric acid-methanol (10∶90) as mobile phase and 213nm as detected wavelengths.Results The contents of the above two alkaloids showed great difference among the 14 patches of tested samples. The content of ephedrine ranged 0.361 %～1.538 %and that of pseudo-ephedrine 0.332～2.087 %. The suggested content limitation of ephedrine should be 1.082 %, and that of psedoephedrine be 1.008 %in the crude Herba Ephedra.Conclusion The established method has been proven to be simple, stable and repeatable, and can be applied for quality control of the crude crude Herba Ephedra and compound prescriptions containing crude Herba Ephedra.

Objective To investigate the prevalence of nutritional risk,undemutrition,overweight/obesi-ty,and apphcation of nutrition support in departments of neurology in 3 metropolitan hospitals in Beijing. Methods Adult patients in the departments of neurology from 3 metropolitan hospitals in Beijing were consecutively en-rolled from March 2005 to March 2006.Nutritional Risk Screening 2002(NRS2002)score≥3 was defined a8 nu-tritian risk,while body mass index(BMI)<18.5 ks/m2 as undemutrition.NRS2002 was performed on the next morning of admission and nutritional support evaluation was performed on the 14th day of admission or discharge day.The relationship between nutrition risk and nutrition support was analyzed.Results A total of 753 patients were enrolled.NRS2002 scoring results were obtained in 461 patients.The prevalence of undemutrition was 4.2%and nutritional risk Was 21.2%.Among them 292 cases in undemutrition category were estimated by semm albu-min<30g/L without accurate BML If the cases without accurate BMl were excluded,the prevalence of undemutri-tion was 5.4%and nutritional risk Was 10.8%.Twenty-three patients(14.4%)with NRS2002≥3 received nutrition support.Thirteen patients(2.2%)with NRS2002<3 received nutrition support.The average PN:EN ratio Wag 1:2.Conclusions A large proportion of inpatients were at nutritional risk or undemutrition in the de-partments of neurology in metropolitan hospitals in Beijing.The application of nutrition support Wag somehow inap-propriate in these patients.Evidence-based guidelines are required to improve this situation