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 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 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 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.

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.

Objective To fabricate cartilage extracellular matrix (ECM) oriented scaffolds and investigate the attachment, proliferation, distribution and orientation of bone marrow mesenchymal stem cells (BMSCs) cultured within the scaffolds in vitro. Methods Cartilage slices were shattered in sterile phosphate-buffered saline (PBS) and the suspension were differentially centrifugated untill the micro- fiber of the cartilage extracellular matrix was disassociated from the residue cartilage fragments. At last the supernatant were centrifugated, the precipitation were collected and were made into 2%-3% suspension. Using unidirectional solidification as a freezing process and freeze-dried method, the cartilage extracellular matrix derived oriented scaffolds was fabricated. The scaffolds were then cross-linked by exposure to ultraviolet radiation and immersion in a carbodiimide solution. By light microscope and scan electron microscope (SEM) observation, histological staining, and biomechanical test, the traits of scaffolds were studied. After being labelled with PKH26 fluorescent dye, rabbit BMSCs were seeded onto the scaffolds. The attachment, proliferation and differentiation of the cells were analyzed using inverted fluorescent microscope. Results The histological staining showed that toluidine blue, safranin O, alcian blue and anti-collagen Ⅱ immunohistochemistry staining of the scaffolds were positive. A perpendicular pore-channel structures which has a diameter of 100 μm were verified by light microscope and SEM analysis. The cell-free scaffolds showed the compression moduli were (2.02±0.02) MPa in the mechanical testing. Inverted fluorescent microscope showed that most of the cells attached to the scaffold. Cells were found to be widely distributed within the scaffold, which acted as a columnar arrangement. The formation of a surface cells layer was found on the surface of the scaffolds which resembled natural cartilage. Coclusion The cartilage extracellular matrix derived oriented scaffolds have promising biological, structural, and mechanical properties.