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1.
Article in Chinese | WPRIM | ID: wpr-870167

ABSTRACT

In recent years, the clinical experts consensuses or guidelines of ankylosing spondylitis (AS)/spondyloarthritis (SpA) have been constantly updated, but to better understand and practice, patient self-participation management is one of the key points to improve the level of diagnosis and treatment. Through questionnaire survey of these patients, we screened out the most concerned issues, and established the AS/SpA patient practice guideline working group with multidisciplinary physicians and patients. Fifteen opinions, as the AS/SpA patient practice guidelines, are proposed in accordance with the relevant principles of the "WHO guidelines development manual" , and with the international normative process.

2.
Article in Chinese | WPRIM | ID: wpr-707498

ABSTRACT

Objective To investigate the mechanism of inducing production of vascular endothelial growth factors (VEGF) by recombinant human S100 calcium binding protein A4 (rhS100A4) in rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs).Methods Synovial tissue was sampled from the patients with rheumatoid arthritis (RA) undergoing knee arthroplasty for in vitro culture of RAFLSs.CCK-8 assay was conducted to detect the effect of rhS100A4 and the effect of its interaction with Rapamycin (Rap),an inhibitor of mammalian rapamycin target 1 (mTORC1) signaling pathway,on the proliferation of RAFLSs.The effects of rhS100A4 and its interaction with Rap on the expression of VEGF in RAFLSs were detected by immunofluorescence.After rhS100A4 and its cooperation with Rap stimulated the conditioned medium (CM)produced by RAFLSs,the effect of CM on formation of lumen in human unbilical vein endothelial cells (HUVECs) in vitro was observed to detect the angiogenic ability of rhS100A4.Western blot was used to detect the effect of rhS100A4 on the phosphorylation of downstream ribosomal protein S6 (S6) in the mTORC1 signaling pathway in RAFLSs and to analyze the effects of rhS100A4 and Rap on phosphorylation of S6 protein and expression of VEGF protein in RAFLSs.Results rhS100A4 promoted cell proliferation and expression of VEGF protein in RAFLSs,and the CM formed by rhS100A4 promoted HUVECs to form blood vessels in vitro.Rap inhibited the above biological effects of rhS100A4,rhS100A4 activated the downstream protein S6 in the mTORC1 signaling pathway in RAFLSs cells to increase their phosphorylation levels.The effects of rhS100A4 on the phosphorylation of S6 protein and on the expression of VEGF protein in RAFLSs were inhibited by Rap.Conclusion rhS10OA4 promotes production of VEGF in RAFLSs by activating the mTORC 1 signaling pathway.

3.
Chinese Journal of Pathophysiology ; (12): 1119-1124, 2017.
Article in Chinese | WPRIM | ID: wpr-612937

ABSTRACT

AIM:To study the expression level of S100 calcium-binding protein A4 (S100A4) in synovial tissue of the knee joint in rheumatoid arthritis (RA) patients and normal persons, and the effect of S100A4 on the angiogenesis induced by rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs).METHODS:The synovial tissue was taken from the knee joint of the RA patients (RA group) and the normal persons (control group).The protein expression of S100A4 and vascular endothelial growth factor (VEGF) in the synovial tissue of the 2 groups was observed by immunohistochemistry.RAFLSs were isolated from synovial tissue of patients with active RA.ELISA was used to detect the effect of S100A4 on the secretion of VEGF by RAFLSs.The effect of S100A4 on the angiogenesis of HUVECs cultured with conditioned medium from RAFLSs was also detected.RESULTS:The protein of S100A4 and VEGF was highly expressed in the synovial tissues of RA group (P<0.05).rhS100A4 significantly stimulated the secretion of VEGF in RAFLSs in a time-and dose-dependent manner (P<0.05).Cultured with conditioned medium from RAFLSs, rhS100A4 significantly promoted HUVECs to form tube-like structures in vitro.CONCLUSION:S100A4 protein is highly expressed in synovial tissue of the knee joint in RA patients, and S100A4 stimulates synovial angiogenesis by promoting RAFLSs to generate VEGF, indicating that S100A4 may be used as a potential target for the treatment of RA.

4.
Article in Chinese | WPRIM | ID: wpr-510976

ABSTRACT

[Objective]To investigate the efficacy of total hip arthroplasty(THA)on the treatment of traumatic arthritis that caused by internal fixation failures of intertrochanteric fractures.[Methods]During January 2009 and March 2016,35 cases of trau?matic arthritis(male:18 cases;female17 cases;49 ~ 86 years old,with an average age of 68.5 years)caused by internal fixation failures or malunion of intertrochanteric fractures,were undergo THA. Among 35 cases,13 cases were performed with the proximal femoral fixation stems,10 cases were with distal fixation stems,and 12 cases were with extended stems.[Results]With 3~65 months follow-up,the hip joint HSS score was elevated from 44.1(31 ~ 65)preoperative to 82.5(58 ~ 94)postoperative without obvious loosening. No postoperative deep infectionwas found. The femoral stems in 2 cases were found to sink 5 mm and 10 mm,respectively. No obvious prosthesis loosening was found. Taken together ,the satisfaction rate of THA on the joint function of traumatic arthritis was 91.4%.[Conclusion]Total hip arthroplasty is recommended as an effective approach for treating traumatic arthritis caused by internal fixation failures of intertrochanteric fractures. Distal fixed prosthesis was recommended due to bone sclerosis or defects of proximal femur. Coupled with emphasis on reconstruction of the greater trochanter ,good therapeutic effects could be achieved.

5.
Article in Chinese | WPRIM | ID: wpr-515365

ABSTRACT

There is a long path to go on the course of Chinese medical supply reform program.With the development of medicare system,the use of Internet + to promote Chinese medical supply reform program and to provide usefulsolutions to the present medicare problems in China could be very difficult.

6.
Article in Chinese | WPRIM | ID: wpr-486651

ABSTRACT

Protein arginine methyltransferases ( PRMTs) play crucial roles in the methylation of a series pro-tein substrates.PRMT5 is a type Ⅱ methyltransferase that symmetrically methylates arginine residues of histone and non-histone substrates, thereby regulating a variety of cellular processes through epigenetic control of target gene expression or post-translational modification of signaling molecules.Recently, accumulated evidence has suggested that PRMT5 may function as an oncogene.This review is aimed to summarize the oncogenic role of PRMT5 and its regulatory mechanisms in tumors.

7.
Article in Chinese | WPRIM | ID: wpr-464260

ABSTRACT

AIM:To conduct the relevance analysis of serum Dickkopf-1 (Dkk-1) and bone mineral density (BMD) in the different ages of female populations.METHODS: The women volunteers (n=100, 20~80 years old) were selected and divided into young group (20~39 years old), middle age group (40~59 years old) and elderly group (60~80 years old).The serum levels of Dkk-1 in the 3 groups of volunteers were detected by ELISA.In the middle age group, 25 people of 45~55 years old were selected and divided into postmenopausal group and premenopausal group to de-tect the serum level of Dkk-1 in the 2 groups of volunteers by ELISA.The BMD was measured by the method of dual energy X-ray absorptiometry.The differences of Dkk-1 expression levels among different ages of female populations, and the rele-vance with BMD were compared.RESULTS:With the increase in age, the serum Dkk-1 expression level increased ( P<0.05), and BMD were reduced (P<0.05).The blood level of Dkk-1 and BMD negatively correlated (P<0.05) in the 3 groups of volunteers.The serum levels of Dkk-1 and BMD had stronger negative correlation in postmenopausal women group than that in premenopausal women group.CONCLUSION:With the increase in age, the expression level of serum Dkk-1 increases and the BMD level decreases, which contribute to a risk of osteoporosis.In the same age range, the postmeno-pausal women express higher level of Dkk-1, and the decreased BMD is more obvious, which contribute to a greater risk of osteoporosis.The increased level of Dkk-1 also inhibits bone formation and promotes bone resorption.It may become a new target for preventing and treating osteoporosis.

8.
Article in Chinese | WPRIM | ID: wpr-462915

ABSTRACT

BACKGROUND:Correlation between subchondral bone and articular cartilage in the process of osteoarthritis has not been fuly elucidated. Degeneration of cartilage is the focus of attention, and the subchondral bone also plays an important role in the process of osteoarthritis. OBJECTIVE: To observe the differences between experimental osteoarthritis models in rabbit knees established by two kinds of surgical methods and two kinds of proteases inducing methods, and to explore the correlation between subchondral bone mass and degeneration of cartilage. METHODS:Thirty-two New Zealand rabbits were randomly and averagely divided into four groups: Hulth group (group A), anterior cruciate ligament transaction group (group B), colagenase type II group (group C) and papain group (group D). The right knees of rabbits were established as osteoarthritis models, and the left knees served as controls. Bone mineral density of the knee joint was evaluated by dual-energy X-ray absorptiometry scanning at 0, 4 and 8 weeks after modeling. The rabbits were sacrificed at 8 weeks after MRI scanning, bilateral knee joints were harvested for general and histological observation. Quantitative analysis was done according to Mankin scores. RESULTS AND CONCLUSION: Bone mineral density of the right knees decreased at 0, 4 and 8 weeks after modeling, and the rank was as folows: group A > group B > group C > group D. MRI scanning showed that the articular cartilage thickness of the medial and lateral femoral condyle on the right knees became thinner compared with the left side, and the rank was as folows: group A < group B < group C < group D. Observation by specimens and pathological slices showed that the articular cartilage degeneration of the surgery groups worsened, group A was the most serious one, and group 1D was the lightest. Both surgery and proteases inducing methods can successfuly establish osteoarthritis models in rabbit knees. Surgery inducing models resemble the advanced or intermediate stage of osteoarthritis, while the proteases inducing models resemble the early stage of osteoarthritis. Degeneration of the articular cartilage and changes of subchondral bone are related in progressive development.

9.
Chinese Journal of Pathophysiology ; (12): 1483-1489, 2015.
Article in Chinese | WPRIM | ID: wpr-477241

ABSTRACT

[ABSTRACT]AIM:Toexploretheeffectoftheelasticmodulusandsizesofliquidcrystal(LC)phasesonosteo-genic differentiation based on OPC/PU composite substrate by mimicking the microenvironment in rat bone mesenchymal stem cells (rBMSCs).METHODS: A series of composite substrates with different elastic modulus were constructed via modulation of LC content in the composites .The surface phase structure was observed by polarized microscopy , and the mechanical property was measured by a universal material testing machine .Furthermore, the laser confocal microscope was employed to observe the spreading , polarization and the cytoskeleton arrangement of the rBMSCs .The proliferation of rBM-SCs was evaluated by CCK-8 assay.The specific mRNA expression of osteogenic differentiation such as collagen Ⅰ, and osteopontin on the composite membranes was detected by real-time PCR.RESULTS:The size and number of LC phase in-creased and the elastic modulus of the composite substrates decreased with the increase of the LC content .The rBMSCs ex-hibited better characteristics of initial adhesion , spreading and proliferation on the OPC 10-PU and OPC30-PU in the early and medium culturing .The rBMSCs displayed higher expression of collagen Ⅰ and osteopontin on the OPC10-PU in the early and medium osteogenic induction , while the high expression of these osteogenic genes occured on the OPC 30-PU and OPC50-PU in later osteogenic induction .The emphasis of genetic expression was switched from collagen Ⅰin the early and medium osteogenic induction to osteopontin in the later stage .CONCLUSION:When the content of LC remained low in the composite substrates , rBMSCs mainly responded to the mechanical stimuli induced by substrate stiffness and exhibited distinguished cellular behaviors;with the increase in the LC content , rBMSCs had strong interactions with LC by sensing the viscoelasticity of LC , probably resulted from the contribution of both substrate stiffness and the viscoelasticity of LC phase .

10.
Article in Chinese | WPRIM | ID: wpr-438955

ABSTRACT

BACKGROUND:Studies have shown that Wnt signaling pathways play an important role in the osteogenic differentiation of mesenchymal stem cells. OBJECTIVE:To review the mechanism and regulation of the Wnt signaling pathways, as wel as Wnt signaling pathway effects on osteogenic differentiation of mesenchymal stem cells. METHODS:A computer-based search of PuMed database and CNKI database from September 1998 to March 2013 was performed to search related articles. The key words of“Wnt, mesenchymal stem cells, Wnt signaling pathways, osteoblastic differentiation, canonical wnt signaling pathway, non-canonical signaling pathway”in English or Chinese were used to search the articles in the title and the abstract. A total of 31 articles were included to review. RESULTS AND CONCLUSION:Wnt signaling pathways play a critical role in the osteogenic differentiation of mesenchymal stem cells. Canonical Wnt signaling pathway, non-canonical Wnt signaling pathway, and their mutli-factors were involved in regulating the proliferation and differentiation of mesenchymal stem cells. The osteogenic differentiation of mesenchymal stem cells can be promoted effectively via specific induction of Wnt signaling pathways. Wnt11, FZD6, sFRP2, sFRP3 and Ror2 expressions increase, while Wnt9a and FZD7 decreases during the regulation. However, the relations of factors in Wnt signaling pathways and how to use the mechanism of Wnt signaling for promoting mesenchymal stem cells faster, more accurate differentiation need further studies.

11.
Article in Chinese | WPRIM | ID: wpr-414830

ABSTRACT

BACKGROUND: Though there were many experiments addressing repairing osteochondral defects before, faulty restoration occurred at coupling interfaces. OBJECTIVE: To investigate the feasibility of repairing of osteochondral composite defects in rabbit knees with animal-origin osteochondral scaffold combined with bone marrow mesenchymal stem cells (BMSCs)/chondrocytes.METHODS: New Zealand white rabbits were randomly divided into the experimental, control and blank groups and prepared for unilateral knee joint osteochondral defects. Animal-origin osteochondral scaffold combined with BMSCs/chondrocytes, animal-origin osteochondral scaffold and no material was implanted to repair the defects in the experimental, control and blank groups, respectively. Healing condition was evaluated by gross observation, hematoxylin-eosin staining, and toluidine blue staining at 4, 8, and 12 weeks after operation. RESULTS AND CONCLUSION: At 12 weeks after operation, gross observation showed the defects were repaired completely without local depression and the regenerated tissues were fused with surrounding tissues in the experimental group. Hematoxylin-eosin staining and toluidine blue staining revealed that there were many new hyaline cartilages in the cartilage defects in which columnar cells were lined well and cartilage lacuna was obviously, also, there were many bony tissues in the bone defects. The regeneration cartilage, the underlying subchondral bone and host bone were coupled completely. The toluidine blue positive rate and histologic scores of the experimental group were superior to those of the control and blank groups (P < 0.05). It is demonstrated that animal-origin osteochondral scaffold combined with BMSCs/chondrocytes is an ideal method to repair defects between cartilage and the underlying subchondral bone.

12.
Article in Chinese | WPRIM | ID: wpr-402495

ABSTRACT

BACKGROUND: With further understanding of deep venous thrombosis(DVT)following total hip replacement,reduction and prevention of DVT has become hot topic in clinical studies.The reports of DVT formation factors remain controversial due to small samples,little statistical significance,confusion of basic experimental and clinical results and lacks of science.OBJECTIVE: To explore the causes and factors for the early DVT following total hip replacement and summarize measures to prevent and treat early DVT to reduce incidence of complications.METHODS: A total of 1780 cases of primary total hip replacement operation were analyzed retrospectively.The statistical indexes included sex,age,body mass,other system disease,previous hip joint operation,anesthesia,operative time,prosthetic fixation,blood transfusion,postoperative functional exercise,antithrombotics,and complication.Standardized database was built and analyzed by SPSS(version 13).Regression analysis was performed using Binary Logistic Regression.RESULTS AND CONCLUSION: Of 1780 cases,136 had DVT.Age,other system diseases,anesthesia,prosthetic fixation,blood transfusion,postoperative functional exercise and antithrombotics were correlated with early DVT(P < 0.05).Old age,hypertension or diabetes,general anesthesia,fixation of bone cement,whole blood transfusion were the risk factors for early DVT following total hip replacement,while postoperative functional exercise and antithrombotics were the protective factors for DVT.The incidence rate of early complications can be reduced by the methods such as dealing with perioperative treatment carefully,effectively controlling the chronic diseases,efficient evaluation before surgery,precise manipulation,and the postoperative prophylactic treatment and nursing.

13.
Article in Chinese | WPRIM | ID: wpr-402457

ABSTRACT

BACKGROUND: The development of cartilage tissue engineering provides novel ideas for treatment of articular cartilage defects and implements construction of tissue-engineered cartilage in vivo.OBJECTIVE: To investigate the feasibility of constructing tissue-engineered osteochondral composite through bone marrow stem cells(BMSCs) cultured on the poly(lactide-co-glycolic acid) (PLGA), which was modified with collagen and cellular growth factors.METHODS: PLGA was made by phase separation technique, composited with collagen Ⅱ, basic fibroblast growth factor, and transforming growth factor-β1. The BMSCs of passage 3 were cultured on the above scaffolds. Thirty-six SD rats were randomly divided into experimental, control, and blank groups. These three groups received implantation of BMSCs composited with growth factors and collagen-PLGA, implantation of BMSCs composited with collagen-PLGA, and implantation of collagen-PLGA into the muscle, respectively. At 4, 8, and 12 weeks after surgery, cell directional differentiation and growth were examined by gross observation, hematoxylin-eosin staining, toluidine blue staining, collagen Ⅱ staining, and scanning electron microscope.RESULTS AND CONCLUSION: Gross observation showed that there were many chondroid tissues in the experimental group and fibrous tissues in the control and black groups. Stainings and electron microscope revealed that many chondroblasts and a few osteoclasts appeared in the composite of the experimental group. Toluidine blue and collagen Ⅱ stainings were positive in the experimental group and negative in the control and blank groups. These findings demonstrate that PLGA modified with collagen had a good cellular compatibility. BMSCs cultured on PLGA, which was modified with collagen and cellular growth factors, can construct the tissue-angineered osteochondral composite in rats.

14.
Chinese Journal of Trauma ; (12): 891-893, 2010.
Article in Chinese | WPRIM | ID: wpr-386547

ABSTRACT

Objective To investigate the clinical value and relating problems in treating atlantoaxial instability by using transpedicular instrumentation with fusion. Methods The study reviewed 18 patients (11 males and 7 females, at age range of 13-82 years, mean 46.5 years) with atlantoaxial instability undergone transpedicular screw internal fixation with bone grafts fusion. There were 15 patients with type Ⅱ odontoid fractures, two with traumatic disruption of transverse atlantal ligament and one with fracture of the anterior ring of C1, all of which were associated with atlantoaxial subluxation or obvious instability. Preoperative JOA score was 6-13 points (average 9.5 points). Results The operation lasted for mean 115 minutes (range 75-180 minutes), with intraoperative blood loss of mean 235 ml (range 130-450 ml). One patient presented intraoperative plexus venous bleeding during removal of lower edge of the posterior arch of atlas and was treated with hemostasis using compession of gelatin sponge. All the patients were followed up for a mean period of 13.5 months (6-38 months), which showed no complications including infection, loosening or breakage of internal fixators or neurovascular injury related to internal fixation. Postoperative JOA score was 12-17 points (average 14.5 points). Reduction and solid fusion of the bone graft were achieved satisfactorily in all patients. Conclusions Posterior transpedicular screw internal fixation with bone grafts fusion can achieve good reduction, reliable fixation, high fusion rate and is a reliable method to manage atlantoaxial instability. Correct selection of the indication, familiarity with local anatomy and mastery of the operation technique are key to a satisfactory curative effect.

15.
Article in Chinese | WPRIM | ID: wpr-280183

ABSTRACT

It is currently reported that extracellular matrix, biological scaffolds, conditions of stress, nutrients and metabolic waste play very important roles in tissue-engineered osteochondral composite. In this paper, we have made a review of their effects on such composite.


Subject(s)
Humans , Cartilage , Chemistry , Physiology , Chondrocytes , Connective Tissue , Extracellular Matrix , Chemistry , Stress, Mechanical , Tissue Engineering , Methods , Tissue Scaffolds , Chemistry
16.
Article in Chinese | WPRIM | ID: wpr-407488

ABSTRACT

BACKGROUND: The structure of nanometer chitosan-sodium/collagen (nano-CS/COL) is similar to that of the extracellular matrix (ECM) in the nanometer level. Whether this can promote the adhesion and growth of bone marrow mesenchymal stem cells (MSCs) and the calcification?OBJECTIVE: To investigate the in vitro histocompatibility of nano-CS/COL. DESIGN: Single sample observation.SETTING: Department of Orthopaedics, First Hospital, Jinan University. MATERIALS: This study was performed at the Experimental Center, First Hospital Affiliated to Jinan University between March 2007 and July 2007. Ten 4-week-old female SD rats, of SPF grade, weighing 200 g, were provided by the Guangdong Provincial Laboratory Center [Permission No. SCXK (yue) 2003-0002]. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Nano-CS/COL METHODS: Bone marrow MSCs were isolated from SD rats and cultured. Cell surface antigen was detected by loss cellanalyticalmethod.Nano-CS/COLscaffold waspreparedbypolyelectrolyte confocallaser-scanning microscopy. The well-grown cells of the third passage were co-cultured in vitro on the nano-CS/COL scaffold. Taking simple nano-CS/COL scaffold material as control, the histocompatibility of scaffold material and cells were comprehensively evaluated by cell adherence rate, growth curve, cell activity and cycle, and scanning electron microscope observation.MAIN OUTCOME MEASURES: ① Identification of cell surface antigen marker after isolation and culture of bone marrow MSCs. ②The histocompatibility of nano-CS/COL material and bone marrow MSCs 2, 4 and 8 days after nano-CS/COL material compounded with cells. ③Determination of adherence rate of cells to nano-CS/COL material. ? Cell circle and activity detected 5 days after nano-CS/COL material compounding with cells. RESULTS: ① Detection results of cell surface antigen marker: The expression of CD29, CD106, CD44, CD34 and CD45 was 90.86%, 73.38%, 82.61%, 0.76% and 0.60%, respectively. ②Histocompatibility of bone marrow MSCs and nano-CS/COL material: It was shown under the scanning electron microscope that nano-CS/COL scaffold presented porous three-dimensional structure, and different sizes of macropoles and interconnected small pores. The interval porosity determined by quality assay was 85%-90%, and aperture averaged 150 μm (range 50 - 300u m). Two days after bone marrow MSCs compounded to nano-CS/COL scaffold, bone marrow MSCs presented globular shape and were scattered; Four days later, bone marrow MSCs presented shuttle shape, extended and anchored on the surface of nano-CS/COL by pseudopods; Eight days later, bone marrow MSCs proliferated and fused each other, and they secreted a lot of extracellular matrix, then which covered most material particles. ③ The adherence rate of bone marrow MSCs to nano-CS/COL: Bone marrow MSCs and nano-CS/COL were co-cultured 2 and 6 hours separately. The adherence rate of bone marrow MSCs was higher to nano-CS/COL scaffold than to simple chitosan scaffold. ④ Comparison of cells and cell cycle between on nano-CS/COL scaffold and on the chitosan scaffold: On the nano-CS/COL scaffold, cell activity was 96.67%, cell cycle at G0-G1 was 90.81%, at G2-M was 0.52% and at S was 8.66%. G2/G1 was 1.81. On the simple chitosan scaffold, cell activity was 95.27%, cell cycle at G0-G1 was 87.14%, at G2-M was 9.69%, and at S was 4.16%. G2/G1 was 1.80.CONCLUSION: Nano-CS/COL scaffold can be used as tissue engineering biomaterials because bone marrow MSCs can well grow on it.

17.
Article in Chinese | WPRIM | ID: wpr-407307

ABSTRACT

BACKGROUND: The skill to culture osteoblasts primarily has been well developed. However, trypsinase can affect membrane protein of osteoblasts if the time of digestion is long. Therefore, it is of great significance to select an ideal method to avoid the damage from trypsinase to cells as possible when culturing osteoblasts.OBJECTIVE: To explore a novel method to isolate and culture SD rat osteoblasts in vitro, and identify the functions of the cells.DESIGN: Observational study.SETTING: Department of Orthopaedics, First Affiliated Hospital of Jinan University.MATERIALS: This experiment was carried out in the Department of Orthopaedics, First Affiliated Hospital of Jinan University from March to May in 2007. Eight SPF 24-hour old SD rats were used in the experiment. The rats, irrespective of gender, were provided by the Experimental Animal Center of Nanfang Medical University. The experimental animals were disposed according to ethical criteria. The main reagents were detailed as follows: collagenase Ⅱ (Sigma Company);trypsin (Sigma Company); alkaline phosphatase (ALP) kit (Nanjing Jiancheng Biological Products Company); SABC-1021(Wuhan Boster Biotechnology Company).METHODS: 24-hour old SD rats were chosen for experiment. The newly born SD rats were sacrificed by anesthesia and the cranial bones of the rats were obtained cleanly, erased completely of the periosteum and cut to blocks of I mm3. The cranial bones were digested by 0.25 % trypsinase for 20 minutes, then by 0. 1% type Ⅱ collagenase for 60 minutes. The digestive time of trypsinase was controlled in the process of digestion to avoid to harm the cells. The liquid was gathered and centrifuged. The cells were cultured in culture flask and were purified by many times adhered.MAIN OUTCOME MEASURES: Morphology observations under the inverted phase contrast microscope, transmission electron microscope, and scanning electron microscope were performed. The phenotype, calcium tuberculation and the expression of alkaline phosphatase were studied with alizarin red staining and modified Gomori Ca-Co assays respectively.The cells were also evaluated with collage Ⅰ immunohistochemical staining.RESULTS: The cultured cells had active proliferation ability. Cells showed multi-angle or fusiform shape. Nucleus was immature and organell was plentiful. Therefore, they had typical morphological characters of osteoblasts. Moreover, they showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase, collage Ⅰ and formation of calcium tuberculations.CONCLUSION: The cells cultured by our modified enzymatic digestion method had typical morphological and biological characteristics of osteoblasts.

18.
Article in Chinese | WPRIM | ID: wpr-408000

ABSTRACT

BACKGROUND:There is a great difference of grade size of macrobead in various joint diseases; therefore, it can be used to determine state of joint diseases initially.OBJECTTVE : To explore the physical properties of synovial fluid nano-particles and their correlations with the occurrence of knee osteoarthritis (KOA).DESIGN: Controlled experimental study based on synovial fluid samples.PARTICIPANTS: A total of 99 synovial fluid samples were collected from normal subjects and KOA patients with various KOA severities. Among them, 41 were normal synovial fluids, 58 were KOA.METHODS: Synovial fluid samples from individuals with and without KOA were obtained. Using the technology of quasi-elastic laser scattering, nano-particle size and its distribution were estimated, and the dynamic/static light scattering spectrometric analyzer allowed the measurement of particles Zeta potentials. A correlation analysis between the particle size, Zeta potentials and the onset of KOA was attempted.MAIN OUTCOME MEASURES:① Grade size and distribution of microsome in synovial fluid;② Zeta potentials and distribution of microsome in synovial fluid; ③ grade size and clinical correlation of microsome in synovial fluid.RESULTS: ① The mean nano-particle diameter in the synovial fluid of KOA patients were significantly greater than those of normal joints [(297±84), (63±23) nm, P < 0.001]. The distribution curve of KOA synovial fluid nano-particle size was normal knee and (-15.84 ±3.34) mV of KOA patients, and there was a significant difference (P < 0.001). This suggestedthat the Zeta potentials in the synovial fluid of KOA patients were significantly greater than those of normal joints. ③ The average particle size and Zeta potential of synovial fluid strongly correlated with the integrity of the joint of KOA (rp =0.797 2,0.631 9, P< 0.01).CONCLUSION: The nano-particle size and Zeta potential of synovial fluid are significantly correlated with the development of KOA, and this can reflect the severity of KOA.

19.
Article in Chinese | WPRIM | ID: wpr-545547

ABSTRACT

[Objective]To explore a method for isolating and culturing bone marrow mesenchymal stem cell(BMSCs) in vitro,and improve the cellular conglutination ability of biomaterial poly-lactide-co-glycolic acid(PLGA),and observer the adhesion of BMSCs cultured on PLGA modified with Ⅱ collagen.[Method]BMSCs were isolated and cultured in vitro.The cell exterior antigen,cell livingness and cell cycle were analyzed by flow cytometry method,and cellular configuration was observed continually under invertd phase-contrast microscope.PLGA was made by phase separation,and was modified with Ⅱ collagen.The third era BMSCs were cultured on PLGA,and its adhesion with biomaterial was observed scan electron microscope.[Result]BMSCs could be isolated and cultured in vitro,and express CD29,CD44,CD106,but not express CD34,CD45.The cell livingness was 88.96%,cells in G0-Glperiod are 90.32%.The cell morphology was spindle.The average diameter of PLGA modified with Ⅱ collagen was 100 ?m,and PLGA has good adhesion with BMSCs.[Conclusion]BMSCs could be cultured stability in vitro for long time,and is good seminal cell of tissue engineering.Ⅱ collagen can improve the cellular adhesion of PLGA,PLGA modified with Ⅱ collagen is the good biomaterial of tissue engineering.

20.
Article in Chinese | WPRIM | ID: wpr-543277

ABSTRACT

[Objective]To discus clinic value of fixation by vertebral pedicle screw system and vertebroplasty using injectable graft for thoracolumbar vertebrae fractures.[Method]Fifteen cases of thoracolumbar vertebrae fractures(7 cases with compress fractures,8 cases with burst fractures) were treated with fixation by vertebral pedicle screw system and vertebroplasty using injectable graft.[Result]The group was followed up for average 9.6 months,no case showed internal fixation device loosening or breaking,nor were chronic lumbar pain seen in this group.No case had lost the anterior height of body of spine.All injected grafts were Abs orbed within 3 months postoperatively.According to Frankels grading,there were 4 cases in Grades B,6 cases in Grade C and 2 cases of Grade D preoperatively,but 3 cases of Grade C,5 cases of Grade D and 4 cases of Grade E postoperatively in 12 cases with incomplete paraplegia,with a statistically significant difference(x~2 =21.000,P=0.000

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