Periosteal autograft from different sources have been used to repair 1.5cm bone defects of radius in 10 rabbits.On the left side,sharp-dissected grafts were implanted and the animals were sacrificed 4,8,14,30 and 60 days after operation.Bone formation was studied with X-ray and histologic technique.The results showed that the way of sharp-dissecting can preserve the periosteum completely,and good bone formation was found in this group.It suggested that the sharp-dissecting of the periosteum is the important key for periosteal graft.

Objective To investigate the bionomics of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. Methods Umbilical arteries, vein and umbilical cord tunica externa were removed, and the remaining tissue (Wharton's jelly) was harvested. To gather MSCs, the umbilical cord was cut into small fragments and digested with 0.075% type Ⅰ collagenase, or the small fragments were cultured with DMEM. The cells derived from Wharton's jelly of umbilical cord were serially subcultivated, the growth curve was drawn, and the identification of living cells was made for studying the cell growth kinetics. The surface antigens were detected with flow cytometry, the cartilage markers were detected by histochemistry and immunohistochemistry, and the expressions of Sox-9 and Col-2A1 mRNA were analyzed by RT-PCR. Results The confluence time of primary culture cells was 3-5d in cells harvested with enzyme digestion, and 10-15d in cells harvested with micro mass. The results of flow cytometry showed that the MSCs derived from umbilical cord expressed CD44 and CD105; the expression of HLA-ABC was positive, while HLA-DPDQDR was negative. There was no significant change on the immunophenotype of umbilical cord MSCs after cryopreservation and thawing. The findings of histochemistry and immunohistochemistry showed that the expression of chondrocyte markers in Wharton's jelly MSCs was weakly positive. The results of RT-PCR showed the chondrocyte markers Sox-9 and Col-2A1 were positively expressed in Wharton's jelly MSCs. Conclusion The MSCs derived from Wharton jelly of human umbilical cord do not differentiate into hematopoietic cells, may express the chondrocyte markers (they are suggested to have characteristics of pre-chondrocytes), and is expected to be a new type of stem cells of tissue engineering cartilage.

Objective The purpose of this study is to observe the property of guided bone regeneration by collagen membrane and investigate the mechanism of guided bone regeneration. Methods 10mm defect of radius was created bilaterally in adult New Zealand rabbits. The experimental group was transplanted by collagen membrane with surface decalcified bone allograft,and the control group was transplanted by surfaced decalcified bone allograft alone.Radiological and histological and immunohistochemical examinations were taken postoperatively. Results In the transplantation area of experimental group,distinctive periosteal reaction and new bone growth occurred remarkably,bone remodeling progressed successfully,and defects healed completely.But in the transplantation area of the control group,new bone growth and mature bone replacement were delay due to occupation of fibrous connective tissue. Conclusions Collagen membrance has an ability to block and guide.In addition,membrane tube can keep endogenic BMP with exogenic BMP greatly concentrating and effectively distributing. The distributive characteristic of BMP exerts an influence on cellular resources and pattern of bone healing.

Objective To develop a method to prepare human articular cartilage derived microcarrier for both rapid propagating chondrocytes and being used as scaffold to support chondrogenesis. Methods Human articular cartilage was crushed into small pieces by muller after lyophilization, and sorted through two different meshes to collect only those specimens measuring 150-200 microns. Then, in turn, the specimens were subjected to 0.25% trypsin at 37 ℃ for 24 hours and 1% Triton X-100 for 72 hours, respectively. The specimens were observed by inverted phase contrast microscopy, and assessed by staining with haematoxylin-eosin, safranin-O (for GAG), as well as by the immunohistochemistry of aggrecan, collagen type Ⅱ. The microcarriers were seeded with human chondrocytes after being irradiated by 60Co. Results Using inverted phasecontrast microscope, the freezing-dry cartilage particles were observed as yellow, different shapes, and their surfaces were uneven, and with many pits. After treating with trypsin and Triton X-100, the microcarriers showed light yellow, without cartilage morphology. The microcarriers became flocculous or like a hairbrush, and the area of contacting surface significant increased. After culture with cartilage cell for 2 hours, lots of spherical chondrocytes adhered to the microcarriers. HE stain of section confirmed that the celluar constituents of the specimens were removed, the specimens stained weakly positive for GAG, negatively for aggrecan, and positively for collagen type Ⅱ, respectively. Conclusion The detergent and trypsin can remove the cellular constituents and knock out the aggrecan from human articular cartilage while maintaining collagen type Ⅱ and GAG, and made the cartilage pieces flocculous or hairbrush-like. The chondrocytes can be well maintained in human articular cartilage derived microcarriers. Human articular cartilage derived microcarriers were prepared successfullly.

[Objective]To analyze factors related with surviving rate and to evaluate effectiveness of the adjuvant chemotherapy in the treatment of osteosareoma.[Method]Eighty-four patients aging from 9 to 47 years(averaged,21 years)were analysed respectively:52 of them were male and 32 were female.The tumors were located at the femur in 42,the tibia in 29,the humerus in 13.There were 22 patients classified as stage ⅡA and 62 patients as ⅡB.The pathological study,of subtype of osteosarcoma revealed that 47 were osteoblastic,11 chondroblastic,19 fibroblastic and 7 other subtypes.There were 46 patients who received the chemotherapy;38 patients without chemotherapy,49 of the 84 patients treated surgically had limb salvage procedures,35 had amputations.Multivariate analsis was done by using the proportional hazards model of Cox,categoric data were analyzed by using the chi-square statistic.[Result]All cases were followed up from 6 to 74 months(with an average of 25.5 months).Cox model analysis showed that age,sex,site,and subtype were not significant prognostic variables in this group of patients;the significant affecting prognosis in patients was Enncking staging and chemotherapy.Chi-square showed significant difference in the higher metastasis rates of lung in the group without chemotherapy than in those with chemotherapy group(P

Periosteal autograft from different sources have been used to repair 1 5cm bone defects of radius in 10 rabbits.On the left side,sharp dissected grafts were implanted and the animals were sacrificed 4,8,14,30 and 60 days after operation.Bone formation was studied with X ray and histologic technique.The results showed that the way of sharp dissecting can preserve the periosteum completely,and good bone formation was found in this group.It suggested that the sharp dissecting of the periosteum is the important key for periosteal graft.

OBJECTIVE:To establish a method for the residues determination of Pb,Cd,Cu,As and Hg in Codonopsis pilo-sula,and evaluate the quality evaluation of C. pilosula of Pingshun County in Shanxi province. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry was adopted with KED scanning model,RF power was 1 550 W,sampling depth was 5.0 mm,plasma gas(argon)flow rate was 16.0 L/min,helium partial pressure was 0.1 mbar,argon gas was 0.6 mbar, the vacuum degree of 5×10-8 mbar,branch turbopump speed was 1 000 hz,sampling cone aperture was 1.0 mm,skimmer aperture was 0.5 mm,the spray chamber temperature was 2.7 ℃,the data collection was repeated 3 times. RESULTS:The linear range was 0-20 ng/ml for Pb(r=0.999 3),0-10 ng/ml for Cd(r=0.998 5),0-250 ng/ml for Cu(r=0.998 8),0-20 ng/ml for As(r=0.999 0) and 0-1.0 ng/ml for Hg(r=0.997 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 95.80%-100.20%(RSD=1.85%,n=6),94.50%-98.00%(RSD=1.26%,n=6),98.52%-102.43%(RSD=1.60%,n=6), 94.90%-98.70%(RSD=2.29%,n=6)and 96.00%-101.00%(RSD=1.84%,n=6);the limits of detection were 0.021 0,0.003 4, 0.043 7,0.115 6 and 0.005 6 ng/kg,respectively. Pb,Cd,Cu,and As were detetcted,and Hg was not detected,the range of total contents was 7.185 2~12.558 0 mg/kg. CONCLUSIONS:The method is simple with good precision,stability and reproducibility, and can be used for the residues determination of Pb,Cd,Cu,As and Hg in C. pilosula;heavy metal residues in C. pilosula in Shanxi Pingshun county does not exceed limit values of national and industry standards.

Objective To evaluate the efficacy of wet compress with herbs for cetuximab correlative erythra. Methods Forty-two patients received radiochemotherapy combined with cetuximab were randomly divided into two groups. The 23 patients in the experimental group received one-week wet compress with 5g/100ml flos lonicerae twice to five times per day. While the 19 patients in the control group were given wet compress with tepid water. The efficacy on day 3 and day 7 were observed. Results The efficacy on erythra was better in the experimental group than that of control group P<0.05. Conclusion The wet compress with flos lonicerae is effective,safe and economical for the treatment of cetuximab correlative erythra,which is deserved to be applied in clinical practice.

BACKGROUND: Elimination of antigenic substances from natural extracellular matrix with the integrity of the tissue structure retained renders the matrix to possess better biocompatibility and provides a cell culture environment close to conditions of the internal environment. Such materials are the primary choice for cell culture scaffold in tissue engineering.OBJECTIVE: To prepare human articular cartilage acellular matrix so as to provide a methodological basis for further study of articular cartilage acellular matrix as cell scaffold materials.DESIGN: A single sample study of bone tissues.SETTING: The experiment was performed in Institute of Orthopedics, General Hospital of PLA, between January and May in 2004. The specimens were obtained from patients requiring joint replacement for femoral neck fracture.MATERIAIS: The experiment was conducted in the Department of Orthopedics, General Hospital of PLA from January to May in 2004. Human articular cartilage specimens were obtained from the femoral head of patients with total hip arthroplasty for femoral neck fracture.METHODS: Totally 10 specimens of fresh articular cartilage(3.5 mm × 4. 5 mm × 2.0 mm) were obtained and freeze-dried for 12 hours. Cartilage acellular matrix was prepared using Triton X-100, Dnase and Rnase and identified by means of hematoxylin-eosin(HE) and safranine O staining and immunohistochemical staining for cartilage proteoglycan.MAIN OUTCOME MEASURES: Histological observation of the articular cartilage acellular matrix and immunohistochemical staining of cartilage proteoglycan.RESULTS: HE and safranine O staining both showed no cellular structure in the matrix with only recesses left by the removed cells. Immunohistochemical staining for cartilage proteoglycan yielded positive results, suggesting the presence of cartilage proteoglycan in the acellular matrix.CONCLUSION: Human articular cartilage acellular matrix can be prepared using the modified four-step procedures with detergent and enzymatic extraction with lyophilization, and the preserved cartilage proteoglycan in the material may retain good pressure resistance.

BACKGROUND:Choose an ideal vector for amplified chondrocytes arouses more and more attention during the construction of epiphyseal cartilage tissue engineering. OBJECTIVE:To explore the feasibility and performance of epiphyseal cartilage tissue engineering scaffold constructed by chitosan and extracellular matrix of cartilage. DESIGN,TIME AND SETTING:An in vitro study was performed at Department of Orthopedics,General Hospital of Chinese PLA from December 2007 to March 2003. MATERIALS:Chitosan(deacetylation:90%;Mr10?105) was provided by Haihui Bioengineering Company,Qingdao;articular cartilage of swine was collected from market. METHODS:Fresh porcine articular cartilages were obtained and shattered in the iso-osmia liquid. After pulverization and gradient centrifugation,3% artilage microfilament suspension was equally mixed with 2% chitosan. Three-dimensional porous scaffolds were fabricated using a simple freeze-drying method. MAIN OUTCOME MEASURES:After the second gradient ethanol treatment,the scaffolds were investigated by histological staining and scanning electron microscopy to measure aperture,porosity,and water absorption rate. MTT test was also done to assess cytotoxicity of the scaffolds. After induced by transforming growth factor-?1(TGF-?1) ,bone marrow mesenchymal stem cells(BMSCs) of rabbits were incubated onto the scaffolds. Cell proliferation and differentiation were analyzed using inverted microscopy and scanning electron microscopy. RESULTS:The three-dimensional porous scaffold had good pore interconnectivity with pore diameter(161?31) ?m,(90.1?1.6) % porosity and(2 361?132) % water absorption rate. The histological staining showed that toluidine blue,safranin O and anti-collagen II immunohistochemistry staining were positive. The intrinsic cytotoxicity assessment of the scaffolds using MTT test showed that the scaffolds had no cytotoxic effect on BMSCs. Most of the BMSCs attached and covered the surface of the scaffolds with matrix secretion. CONCLUSION:The three-dimensional porous scaffold constructed by extracellular matrix of cartilage and chitosan has good pore diameter and porosity,non-toxicity and good biocompatibility,so it is a suitable scaffold for epiphyseal cartilage tissue engineering.