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1.
Article in Chinese | WPRIM | ID: wpr-921749

ABSTRACT

Hot melt pressure-sensitive adhesive(HMPSA) has broad application potential in the field of traditional Chinese medicine(TCM) plasters due to its high drug loading, weak skin irritation, satisfactory adhesion, etc. compared with rubber plasters.However, the structure of HMPSA is prone to suffer from the damage caused by volatile oils in TCM plasters. In view of this, a kind of HMPSA with a stable structure was prepared by physical blending of DINCH, polypropylene wax and liquid rubber(LIR) in the present study, which is denoted as DPL. The dosage of cinnamon volatile oil(CVO), the model drug, was selected with viscosity, softening point and cohesion as evaluation indexes. The interaction between DPL and HMPSA was investigated by Fourier transform infrared spectroscopy(FT-IR) and differential scanning calorimetry(DSC). The compatibility of HMPSA with CVO and its transdermal ability were studied by in vitro transdermal test, adhesion, scanning electron microscopy( SEM) and rheological evaluation. The results showed that 5% CVO began to damage the structure of HMPSA. The initial adhesion and holding adhesion of DPL-modified HMPSA(DPL-HMPSA) were not significantly changed compared with those of HMPSA, whereas the 180° peel strength was decreased. FI-IR unraveled that DPL formed the n-π conjugated system with styrene-isoprene-styrene block copolymer(SIS), and there was no significant difference in the glass transition temperature according to DSC results, which indicated the good compatibility of DPL with HMPSA. With 5% CVO loaded, the drug content of DPL-HMPSA was 1. 14 times higher than that of HMPSA, and the decrease rate of drug content in DPL-HMPSA was 16% lower than that in HMPSA after 3 months. SEM demonstrated that CVO did not cause obvious structural damage to DPL-HMPSA. Rheological evaluation revealed that the storage modulus and loss factor of DPL-HMPSA were higher than those of HMPSA, and the cohesion was also stronger. The percutaneous penetration rate of cinnamaldehyde in DPL-HMPSA was 2. 25 times that of HMPSA. In conclusion, DPL-HMPSA had more stable structure, better compatibility with CVO, and higher in vitro transdermal efficiency of cinnamaldehyde than before the modification. This study can provide reference for the mitigation of the matrix structure damage caused by volatile oil components in TCM plasters and the enhancement of the content and in vitro transdermal rate of drug.


Subject(s)
Adhesives , Administration, Cutaneous , Cinnamomum zeylanicum , Oils, Volatile , Spectroscopy, Fourier Transform Infrared
2.
Article in Chinese | WPRIM | ID: wpr-691543

ABSTRACT

Objective:To repaire the posterior tooth defects in the elderly people with CEREC chairside CAD/CAM restorations,and to evaluate the clinical curative effect 1 year after treatment.Methods:A total of 52 posterior teeth from 48 aged patients were selected.Among them,24 cases were onlays,18 cases were inlays,and 10 cases were crowns.The inlays/onlays/crowns were designed and manufactured by CEREC chairside CAD/CAM system,and then bonded with resin cement.After 12 months,the patients were followed up.The teeth with restorations were assessed by using the modified USPHS criteria including restorations,tooth,and periodontal.At the same time,the patient's satisfaction was evaluated.Results:In all of the 52 restorations,the success rates of inlay,onlay and crown were 94.4%,91.7% and 80.0%,respectively;among them,one inlay was fractured,one onlay did not close the edge,one onlay's edge integrity was mild defect,one teeth was fractured,one crown's adjacent relationship was not close,and one crown appeared papillitis.Conclusion:The restorations manufactured by CEREC chairside CAD/ CAM system have a good short-term clinical curative effect.It is an effective way to repair the posterior tooth defects in the elderly people.

3.
Article in English | WPRIM | ID: wpr-776641

ABSTRACT

BACKGROUND@#Microvascular complications in type 2 diabetes (T2DM), including diabatic retinopathy (DR), diabetic kidney disease (DKD), diabetic peripheral neuropathy (DPN) are the leading causes of visual loss, end-stage renal disease or amputation, while the current therapies are still unsatisfactory. Chinese medicine (CM) has been widely used for treating diabetic mellitus. However, most of the previous studies focused on the single complication. The role of CM treatment in T2DM patients with 2 or multiple microvascular complications is not clear.@*OBJECTIVE@#To appraise the curative effect of CM in T2DM patients with 2 or multiple microvascular complications, and to compare the effects of stationary treatment and individualized treatment in T2DM patients with microvascular complications.@*METHODS@#This trial will be an 8-center, randomized, controlled study with 8 parallel groups. A total of 432 patients will be randomized to 8 groups: DR study group (32 cases) and a corresponding control group (32 cases), DR+DKD study group (64 cases) and a corresponding control group (64 cases), DR+DPN study group (64 cases) and a corresponding control group (64 cases), DR+DKD+DPN study group (56 cases) and a corresponding control group (56 cases). The control group will receive stationary treatment, and the study group will receive individualized treatment based on CM syndrome differentiation in addition to stationary treatment. The study duration will be 50 weeks, comprising a 2-week run-in period, 24 weeks of intervention, and 24 weeks of follow-up. The outcomes will assess efficacy of treatment, improvement in CM symptoms, safety assessments, adherence to the treatment, and adverse events.@*CONCLUSION@#This study will provide evidence of evidence-based medicine for CM treatment in two or multiple microvascular complications caused by T2DM. (Registration No. ChiCTR-IPR-15007072).


Subject(s)
Diabetes Mellitus, Type 2 , Drug Therapy , Diabetic Angiopathies , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Humans , Medicine, Chinese Traditional , Multicenter Studies as Topic , Outcome Assessment, Health Care , Randomized Controlled Trials as Topic
4.
Article in Chinese | WPRIM | ID: wpr-635593

ABSTRACT

Background Seed cells and scaffold material are the important aspects of corneal tissue engineering research.Adipose-derived stem cells(ASCs) are becoming the focus of seed cells research because of their wide source,powerful proliferation and differentiation abilities.As biodegradable polymer,polylactic-co-glycolic acid(PLGA) has successfully build multiple tissues and organs.Objective Present study was to ascertain the biological characteristics of the rabbit ASCs and their biocompatibility with PLGA scaffold in vitro and to provide groundwork for further study on the reconstruction of tissue engineered corneal stroma.Methods Adipose cells were isolated from lipoaspirate of New Zealand white rabbit using collagenase Ι.The cells were cultured and passaged.The generation 4 cells were inoculated to culture plate with 6 holes at the density of 3×104/cm2,3×104/cm2,3×106/cm2 respectively and cultivated in ossification inducing medium,lipoblast inducing medium and chondroblast inducing medium to identify the characteristics of the cells.The multilineage differentiated cells were identified by alizarin red staining,oil red O staining and immunoinfluorescene technique.The generation 4 cells were re-suspended with DiO influorescence fluid at the density of 1×107/ml and seeded on PLGA scaffold to fabricate cell-PLGA constructs.Quantitative analysis of cell proliferation on PLGA was detected by Hoechst DNA assay.The attachment and growth of adipose-derived stem cells on the scaffold were observed under the scanning electron microscope(SEM) and confocal microscopy in 1 day,3,7 days after seeding for the evaluation of biocompatibility between cells and PLGA.Results Primarily cultured cells reached 80%-90% confluence after 7-8 days with the fibroblast-like appearance.Adipose-derived stem cells of rabbits differentiated into osteoblast,adipocyte and chondroblast successfully,showing the positive stain for alizarin red staining,oil red O staining and immunoinfluorescene technique respectively.Proliferation of cells on PLGA scaffold went into plateau phase at 7 days after culture.SEM and confocal microscopy revealed the well-attached,spread cells along the scaffold and abundant excellular matrix both on the surface and interior pore of scaffold.Conclusion Cultured rabbit adipose cells have the ability of potential multilineage differentiation and good biocompatibility with PLGA scaffold,which could be used to construction of tissue engineered corneal stroma.

5.
Article in Chinese | WPRIM | ID: wpr-591485

ABSTRACT

Objective To explore the cultural method of lingual keratinocytes and make preparations for further investigation in using lingual keratinocytes as a new choice of seed cells for the reconstruction of tissue engineered corneal epithelium.Methods Keratinocytes were enzymatically dissociated from lingual tissue specimens using 0.25%disepase and 0.05%trypsin.The biological characteristics of the cells were observed through phase microscope.Meanwhile.1ingual kemtinoeytes were immtmohistochemically labeled with broad spectrum keratin antibody (AE1/AF3).Results The cells revealed the form of "road metal" and could be passed for 3 or 4 generations.Immunohistochemistry indicated that lingual keratinocytes were positive for AE1/AE3 staining.Condusion Lingual kemtinocytes of rabbits can be cultured with KSFM in vitm and magnitude quantity can be attained,laying a favorable foundation for lingual keratinocytes as a new choice of seed cells for corneal epithelium reconstruction with tissue engineering.

6.
Article in Chinese | WPRIM | ID: wpr-404635

ABSTRACT

BACKGROUND: The application of adipose derived stromal cells to tissue engineering has been more and more popular around the world. Compatibility of scaffold material is the key point for its further research.OBJECTIVE: To determine the growth rules of rabbit adipose-derived stromal cells with polylactic-co-glycolic acid (PLGA).DESIGN, TIME AND SETTING: An in vitro study was performed at the Institute of Ophthalmology, Otolaryngology Hospital,Fudan University and Shanghai Tissue Engineering Center from September 2007 to March 2009.MATERIALS: Six female New Zealand rabbits aged six months were used for extraction of adipose-derived stromal cells. PLGA was provided by Sigma, USA.METHODS: Adipose tissue was harvested from the nape fat pad of the rabbits following anesthesia. Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 10% volume fraction of fetal bovine serum. Cells were passaged when 80% was confluent. The fourth passages of cells were utilized for the study. PLGA consisted of polylactic acid and polyglycolic acid as the ratio of 7:3, and the relative molecular mass was 104900.MAIN OUTCOME MEASURES: Initially, the adherent rate of cells to scaffold was detected. After one week co-culture, the scaffold bearing adipose-derived stromal cells labeled with Dio agent were investigated by fluorescence inverse microscope,scanning electron microscopy and laser scanning microscope.RESULTS: The best adherent rate of adipose-derived stromal cells with PLGA reached 99%. After one-week-incubation in vitro,cells of fiber-shaped or ovule-shaped proliferated well and exhibited stratified growth on the surface of PLGA scaffold. In addition,it also secreted visible extracellular matrices, which could be examined by scanning electron microscopic examination.Meanwhile, the adipose-derived stromal cells grew well and distributed equably inside the PLGA in terms of the investigation with laser scanning microscope.CONCLUSION: The compatibility of adipose-derived stromal cells to PLGA in vitro was well.

7.
Chinese Journal of Surgery ; (12): 847-850, 2008.
Article in Chinese | WPRIM | ID: wpr-258401

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of human caudal-related homeobox 2 (Cdx2) gene expression on human gastric carcinoma cell line MGC-803.</p><p><b>METHODS</b>pCMV-Cdx2-HA eukaryotic expression plasmid was constructed by using DNA recombinant method. The MGC-803 cells were divided into 4 groups: non-transfected, transfected with pCMV-HA, transfected with pCMV-GAPDH-HA, transfected with pCMV-Cdx2-HA. Cdx2 gene and its protein expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot techniques respectively. The effects of Cdx2 overexpression on the growth of MGC-803 cells in vitro was assessed by measuring MTT, flow cytometry and transmission electron microscopy analysis.</p><p><b>RESULTS</b>In MGC-803 cells transfected with pCMV-Cdx2-HA, cell proliferation was significantly suppressed, the cells was ultrastructurally destroyed, cell cycle progression was blocked in G0/G1 and apoptosis rate was higher (P<0.05) in comparison with non-transfected cells or cells transfected with empty vector.</p><p><b>CONCLUSION</b>It indicated that Cdx2 gene can suppress the growth of gastric carcinoma and may save as a novel therapeutic target.</p>


Subject(s)
Apoptosis , Cell Cycle , Cell Line, Tumor , Genetic Vectors , Homeodomain Proteins , Genetics , Metabolism , Humans , Plasmids , Genetics , RNA, Messenger , Genetics , Stomach Neoplasms , Metabolism , Pathology , Transfection
8.
Chinese Journal of Urology ; (12): 368-372, 2008.
Article in Chinese | WPRIM | ID: wpr-400496

ABSTRACT

Objective To investigate the feasibility of replacing urinary epithelial cells with oral keratinocytes by being seeded on bladder acellular matrix graft(BAMG). Methods Twenty-four male rabbits were randomly divided into 2 groups:experimental group and control group.A length of 2.0 cm and width of 0.8 cm penile urethral mucosal defect was induced in the anterior urethra 2.0 cm awav from the urinary meatus in all the rabbits.Oral keratinocytes were isolated from a small buecal mucosa(1.0 cm×0.4 cm)of the 12 rabbits in experimental group and seeded onto a culture dish with a feeder layer of 3T3 mouse fibroblasts inhibited by mitomycin(i3T3).Passage 2 oraI keratinocytes cuItured with i3T3 were expanded and seeded onto sterilized BAMG(2.2 cm×1.0 cm)to repair the de-fects of the urethra.Urethroplasty was performed with BAMG with no cell seeding in the controlgroup.Catheter examination and retrograde urethI ography was done in 1,2 and 6 months after graft-ing.The urethral grafts were harvested and analyzed by histological staining. Results Oral kerati-nocytes seeded onto a feeder layer of i3T3 had better morphous and amplification capability.The corn-patibility of the compound graft was assessed by HE staining and scanning electron microscope.Twelve rabbits in the experimental group could void without difficulty.Catheter examination and ret-rograde urethrogram revealed that a wide urethral caliber was maintained with no sign of strictures and fistula formation.Histological examination showed the grafted urethra was covered with smooth mu- cosa with no strictures at 1,2 and 6 months.The oral keratinocytes of the graft still existed even 6 months after grafting.On contrast,gross and urethrogram demonstrated urethral stricture in the con-trol group.And inflammatory reactions could be found in the area of the stricture through light micro-scope. Conclusions Oral keratinocytes have good compatibility with BAMG and the compound graft could be used for urethral reconstruction in the animal model.

9.
Article in Chinese | WPRIM | ID: wpr-314219

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the feasibility of human bone marrow mesenchymal stem cells (hBMSCs) in vitro differentiation into vascular smooth muscle cells with induction of platelet-derived growth Factor BB (PDGF-BB).</p><p><b>METHODS</b>Bone marrow mesenchymal stem cells of adult healthy donors were separated from iliac crest aspiration and expanded in DMEM-LG medium. Cells at passage 1 were transferred to EGM-2 medium containing PDGF-BB (20 ng/ml) and cultured for 14 days. The expression of SM alpha-actin, SM calponin, SMMHC and SM 22alpha were detected by immunofluorescence and observed with fluorescence microscope. mRNA expression of SMalpha-actin, SM calponin, SMMHC as well as SM 22alpha was analyzed by RT-PCR. The method of Western-Blot was applied to determine protein expression of SM 22alpha. Cells with induction were observed for the expression of SM alpha-actin,SM calponin,SMMHC by FACs analysis.</p><p><b>RESULTS</b>With the induction of PDGF-BB, the morphology of cells changed to a spindle fibroblastic appearance. By fluorescence microscope observation, expression of SM alpha-actin, SM calponin and SMMHC was found intracellularly in PDGF-BB treated hBMSCs at 14 days. Western-Blot detection confirmed SM 22alpha expression by 14 days induction. RT-PCR of characteristic vascular smooth muscle cells related genes, such as SM alpha-actin, SM calponin, SMMHC and SM 22alpha revealed differentiation of vascular smooth muscle cells phenotype in monolayer culture upon stimulation with PDGF-BB for 14 days. The positive expression of SM alpha-actin, SM calponin and SMMHC in induced cells was significantly higher than that in non-induced cells (P < 0.05, n=3).</p><p><b>CONCLUSION</b>These results suggested hBMSCs could be differentiated into vascular smooth muscle cell phenotype with PDGF-BB induction in vitro.</p>


Subject(s)
Adult , Bone Marrow Cells , Cell Biology , Cell Differentiation , Cells, Cultured , Humans , Mesenchymal Stem Cells , Cell Biology , Muscle, Smooth, Vascular , Cell Biology , Platelet-Derived Growth Factor , Pharmacology , Proto-Oncogene Proteins c-sis , Tissue Engineering , Methods
10.
Article in Chinese | WPRIM | ID: wpr-640640

ABSTRACT

Objective To explore the cultural method of oral keratinocytes and make preparations for further investigation in using oral keratinocytes as a new choice of seed cells for the reconstruction of tissue-engineered urethra. Methods Oral keratinocytes of rabbits were isolated in vitro and seeded onto a culture dish with a feeder layer of 3T3 mouse fibroblasts inhibited by mitomycin(i3T3) or a culture dish without i3T3 respectively. Cell morphology was observed and cell growth was detected at intervals.Meanwhile,oral keratinocytes obtained from in vitro culture were performed immunofluorescence staining with broad-spectrum keratin antibody(AE1/AE3) and keratin 19 antibody(K19).The percentage of positive cells of passage 2 reactive to AE1/AE3 was assessed by flow cytometry. ResultsOral keratinocytes seeded onto a feeder layer of i3T3 exhibited finer morphous,better amplification capability,and could be passed for 7 or 8 generations.However,those cultured without i3T3 took on various morphous and could only be subcultured 2 generations before ageing.It was indicated by immunofluorescence staining that oral keratinocytes obtained from in vitro culture were positive for AE1/AE3 staining and 40% were positive for K19 staining.The result of flow cytometry revealed that the amout of positive keratinocytes reactive to AE1/AE3 was more than 95% of total cellular score. Conclusion Oral keratinocytes of rabbits can be cocultured with i3T3 in vitro and magnitude quantity can be attained,laying a favourable foundation for oral keratinocytes as a new choice of seed cells for urethral reconstruction with tissue engineering.

11.
Article in Chinese | WPRIM | ID: wpr-685161

ABSTRACT

Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

12.
Article in Chinese | WPRIM | ID: wpr-544197

ABSTRACT

Objective To study the method of obtaining a large number of dendritic cells (DC). To study the specific cytotoxicity T lymphocyte (CTL) effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor (GM-CSF) and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-? had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54, MHC-Ⅱ and CD86 molecules than control group (P

13.
Chinese Journal of Hematology ; (12): 122-125, 2003.
Article in Chinese | WPRIM | ID: wpr-354905

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of stromal cell derived factor-1alpha(SDF-1alpha) expression and its receptor CXCR4 on the biological behavior of multiple myeloma (MM) cells and on the expression of soluble intercellular adhesion molecule 1 (ICAM-1).</p><p><b>METHODS</b>FACS analysis was used to study the expression of ICAM-1 (CD(54)) and CXCR4 on the surface of MM cells. Chemotaxis assay through transwell bore polycaronate and ELISA assay were employed to monitor the soluble ICAM-1 level.</p><p><b>RESULTS</b>(1) Fresh MM cells expressed variable levels of functional CXCR4 [(50.4 +/- 27.3)%], which was correlated with the in vitro ability of transwell migration of MM cells [(23.6 +/- 17.2)%, P < 0.01]. (2) SDF-1alpha could up-regulate the expression of ICAM-1 on MM cells. Furthermore, the serum level of sICAM-1 was correlated with the expression of CXCR4 on MM cells.</p><p><b>CONCLUSION</b>SDF-1alpha/CXCR4 plays an important role on the biological behavior of MM cells via mediating the effect of adhesion molecules.</p>


Subject(s)
Adult , Aged , Cell Movement , Chemokine CXCL12 , Chemokines, CXC , Female , Humans , Intercellular Adhesion Molecule-1 , Male , Middle Aged , Monocytes , Metabolism , Pathology , Multiple Myeloma , Metabolism , Pathology , Receptors, CXCR4 , Physiology , Tumor Cells, Cultured , Up-Regulation
14.
Article in Chinese | WPRIM | ID: wpr-541905

ABSTRACT

Objective:To study the immune response to murine pancreatic carcinoma by bone marrow-derived dendritic cells(BM-DCs) modified with interleukin-23 after acquiring antigen from apoptotic pancreatic carcinoma cell.Methods:The murine IL-23 cDNA was sub-cloned into dual-expression vector.DCs were pulsed with apoptotic tumor cell antigen after transduced with interleukin-23 gene.The immune preventative and immunotherapeutic effects of DC vaccines on mice with pancreatic cancer were assessed.Results:IL-23 protein could apparently increase the antigen presenting ability of DC.After the vaccination of DC vaccines.IFN-? production in treatment group were significantly more than that of control group(P

15.
Article in Chinese | WPRIM | ID: wpr-411390

ABSTRACT

ObjectiveTo study the surgical treatment of large bowel obstruction caused by colorectal carcinoma.MethodsRetrospective analysis of the experience of surgical treatment for the large bowel obstruction caused by colorectal carcinoma in 52 patients from 1995 to 1999 was performed. ResultsIn the 52 patients, the right hemnicolectomy was performed in 9 patients; left hemicolectomy with proximal colon fistulization was performed in 37patients;transvercolectomy was performed in 4 patients; sigmoid fistulization was performed in 2 patients for their rectum carcinoma couldn't be resected. The postoperative complication rate was 15.3%(8/52),and perioperative mortality rate was 3.8%(2/52). Conclusion More attention should be paid to large bowel obstruction caused by colorectal carcinoma. Selecting rational colectomy and appropriate perioperative management is important for reducing both complication and mortality rate.

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