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OBJECTIVE@#To investigate the changes in the expression of voltage-gated potassium channel subunit KCNA2 in the dorsal root ganglion (DRG) neurons of rats with osteoarthritis (OA) pain induced by sodium monoiodoacetate and explore the mechanism.@*METHODS@#A total of 156 adult male Sprague-Dawley rats were randomly divided into blank control group, saline group and intra-articular monoiodoacetate injection-induced OA group. The paw withdrawal mechanical threshold (PWMT) was measured before and at 1, 2, 4, and 6 weeks after monoiodoacetate injection. At 4 weeks after the injection, the pathological changes in the knee joints were analyzed using HE staining and Safranin O-Fast Green staining, and the expression of activating transcription factor 3 (ATF-3) and inducible nitric oxide synthase (iNOS) in the DRG neurons were detected by immunofluorescence staining. The expression of mRNA in the DRG neurons was detected by RT-qPCR at 1, 2, 4 and 6 weeks after the injection. The expression of KCNA2 in the DRG was measured by Western blotting, and the methylation level of promoter region was measured by MSPCR at 4 weeks after the injection.@*RESULTS@#The PWMT of the rats in OA group was significantly decreased at 2, 4, and 6 weeks after the injection as compared with the baseline ( < 0.05 or < 0.001) as well as the control group ( < 0.05 or < 0.001). Four weeks after the intra-articular injection, fractures and defects on the surface of the articular cartilage, bone hyperplasia, and blurred tidal line were observed in the rats in OA group, but no obvious pathological changes were detected in the control or saline groups. Compared with those in the control group, the expressions of ATF-3 and iNOS were significantly increased ( < 0.01) at 4 weeks after injection; the expression of mRNA at 2, 4 and 6 weeks and the expression of KCNA2 protein at 4 weeks were all significantly decreased ( < 0.05 or < 0.01), and the methylation level of gene was significantly increased at 4 weeks after the injection in OA group ( < 0.01).@*CONCLUSIONS@#The expression of KCNA2 is decreased in the DRG neurons of rats with OA pain likely as a result of enhanced methylation of promoter region.
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Animais , Masculino , Ratos , Modelos Animais de Doenças , Gânglios Espinais , Articulação do Joelho , Metabolismo , Osteoartrite , Metabolismo , Dor , Metabolismo , Regiões Promotoras Genéticas , Ratos Sprague-DawleyRESUMO
Objective To compare the operative and conservative treatments for senile dens fractures of Anderson-D Alonzo type Ⅱ or Ⅲ using Meta-analysis.Methods A literature search was conducted in PubMed,Embase,Web of Science,Cochrane Library,Wanfang Data and CNKI for studies on senile dens fractures of Anderson-D Alonzo type Ⅱ or Ⅲ from the earliest records through June 2016.The relative studies identified were screened by 2 independent authors.The quality of these articles was evaluated using modified Newcastle-Ottawa scale,and the meta-analysis was conducted using Review Manager 5.3.Results A total of 22 articles were brought into this Meta-analysis.The union rate was significantly higher in the operative group than in the conservative group[OR =0.30,95％ CI(0.20,0.44),P ＜ 0.001];the mortality in the operative group was significantly lower than in the conservative group[OR =0.61,95％ CI (0.39,0.96),P=0.03];the complication rate was similar in both groups[OR=1.09,95％ CI(0.76,1.57),P ＜ 0.46].The heterogeneity of all the 3 indexes was low.In the subgroup analysis,the union rate was significantly higher in posterior operations than in anterior operations or conservative treatments (P ＜ 0.05).Conclusion For the elderly patients with dens fracture of type Ⅱ or Ⅲ who can tolerate surgery,operative treatment may be more suitable because it can lead to much better prognosis.
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Aim To determine the effective compo-nents of Semen Ziziphi Spinosae for sedative-hypnotic and its mechanism. Methods The extraction of Se-men Ziziphi Spinosae and the rat brain homogenates were prepared. High concentrations of Diazepam com-petitively replaced the ligand compounds of Semen Ziz-iphi Spinosae combining BDZ receptor in brain tissue, and all the compounds with sedative and hypnotic effects were collected and identified by HPLC and LC-MS technique, as the compounds extracted from the brain tissue were administered with Semen Ziziphi Spi-nosae. The brain tissue was administered with Diaze-pam, and with Semen Ziziphi Spinosae and Diazepam. Results The HPLC chromatograms show that the peak time of BDZ receptor ligand compounds was 2. 71 min and 46. 87min, when compared with Diazepam. And the LC-MS chromatograms display the relative molecu-lar weight of the ligand compounds was 274. 28 m/z, 453. 34 m/z,496. 34 m/z and 608. 38 m/z respective-ly. According to the fingerprint of Semen Ziziphi Spi-nosae, these compounds may be fatty acid substances and lupine pill triterpene compounds. Conclusions On the basis of the principle of receptor ligand bind-ing, we established a way to quickly analyze and iden-tify the role of natural products in the same drug target compounds. The method not only can clearly define the effective components of natural products, but also clar-ify the mechanism of action of the compounds. The ac-tive ingredient of calm hypnosis in Semen Ziziphi Spi-nosae may be fatty acid substances Palmitic acid ( C16 H32 O2 ) and lupine pill triterpene compounds Alphitolic acid( C30 H48 O4 ) and Spinosin( C28 H32 O15 ) . They exert their sedative and hypnotic effects by combining with BDZ receptor, and the research has laid a theoretical foundation for the further study about mechanism of Se-men Ziziphi Spinosae.
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BACKGROUND: Poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) is a novel scaffold made by solvent casting/particulate leaching procedure, composed of polyhydroxybutyrate and polyhyclroxyvalerate at certain ratio, which has good biocompatibility as well as high intensity and modulus. It has three-dimensional porous net structure and good biodegradation. OBJECTIVE: To evaluate the biocompatibility between copolymers of PHBV and canine bone marrow mesenchymal stem cells(BMSCs).DESIGN, TIME AND SETTING: In vitro comparative observation. The study was performed at the Laboratory of Histology and Embryology, Sun Yat-sen University between June 2003 and March 2004.MATERIALS: PHBV scaffold, film porosity > 85% and 100-350 μ m aperture size.METHODS: Canine BMSCs were isolated and cultured. The 3-4 passage cells were seeded onto the PHBV films and three-dimensional foam scaffold. Cells cultured alone served as control.MAIN OUTCOME MEASURES: The seeded cells were observed under inverted microscope; at 1, 2, 3 weeks after seeding, the BMSCs were treated with 4% paraformaldehyde and stained with hematoxylin-eosin (HE); The protein content in seeded cells was determined by bicinchoninic acid assay (BCA), and the content of DNA was quantified using Hoechst33258 assay at 5, 10, 14 days after culture.RESULTS: Inverted microscopic observation showed that the PHBV fibers were fairly thick with weak lucency, and the fibers were hardly detectable under contrast phase microscope. Majority of cells attached onto the PHBV films 2 hours after seeding, and extended well in a spindle shape at 3 days. One week after culture, 2 PHBV were fixed, and BMSCs proliferation was observed after HE staining. At two weeks, cells continued to proliferate and densely covered the PHBV film. The cells grew in the three-dimensional pores, connected at 1 week, extended at 3 weeks, secreting a large amount of material around cells. Cell proliferation did not change much at 3 weeks compared with 2 weeks, and there was no significant difference in DNA and protein contents between control and PHBV groups (P > 0.05).CONCLUSION: As a kind of tissue-engineered scaffold material for BMSCs, PHBV displays good biocompatibility.
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BACKGROUND:Bone marrow stromal stem cells(BMSCs)do not allow for single differentiation of chondrocytes due to their multi-directional differentiation,bone morphogenetic protein secreted from osteoblasts affect the non-differentiated precursor cells and promote their osteoblast differentiations,while those differentiated cells are bound to form tissues.OBJECTIVE:To in vitro induce canine BMSCs differentiate into chondrocytes,and to investigate the method and conditions of chondrocyte differentiation in vitro.DESIGN,TIME AND SETTING:Single sample observation was performed in the Laboratory of Tissue Engineering,Sun Yat-sen University between March 2005 and January 2006.MATERIALS:One male dog,aged 4 months,was involved to harvest BMSCs from the rib.METHODS:Rib BMSCs extracted from bone marrow of 2.0-3.0 mL were cultured in vitro. When cells reached a confluence at 8-11 days,trypsinization was conducted and then halted with L-DMEM synthesis culture solution containing 10% fetal bovine serum. Cellular suspension was collected and centrifuged,cells were rssuspended and incubated at a ratio of 1:3. The third generation of cells were cultured and amplified,10 μg/L basic fibroblast growth factor 2 mL was added to replenish culture medium twice,then 1 mg/L transforming growth factor β1 of 2 mL was applied to induce BMSCs differentiation into chondrocytes.MAIN OUTCOME MEASURES:Toluidine blue and alcian blue stains were applied to determine cartilage matdx secretion,immunohistochemistTy was used for the detection of cartilage specific Ⅱ collagen expression.RESULTS:After BMSCs were primarily cultured and subcultured in vitro,they were shown to grow well at the fourth generation,those induced by basic fibroblast growth factor and transforming growth factor β1 were positive for toluidine blue and aician blue staining;immunohistochemistry showed a positive outcome for type Ⅱ collagen,indicating the induced BMSCs exhibited chondrocyte's characteristics.CONCLUSION:Utilizing basic fibroblast growth factor and transforming growth factor β1,the induced canine BMSCs could differentiate into chondrocytes,which is considered as an ideal seed cells for cartilage tissue engineering.
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Objective To explore the feasibility of building tissue engineered cartilage by bone marrow stromal cells and pbotografting modified copolymers of 3-hydroxybutymte and 3-hydroxyvalerate.Methods Sheep BMSCs were seeded in three-dimensional photografting modified PHBV scaffoids.Twenty-four hours later.composites were cultured with ehondrogenically inductive medium(DMEM)containing TGF-B(10 ng/m1),IGF-1(150 ng/m1)and 20% fetal bovine serum.Three weeks later,the constructs were evaluated by scanning electron microscopy(SEM)and light microscopy with alcian blue,safrine 0 and type Ⅱ collage immunohistochemical staining.GAG contents of constructs were determined by DMB(1,9-dimethylmethylene blue)binding assay at weekly intervals up to 3 weeks.The composites were implanted subcutaneously in sheep abedoml and were evaluated macroscopically and bistologically at 4 weeks postoperatively.Results SEM photograph showed.after one week culture,cell morphology changed from fibroblast-like elongated spindle to the flat rounded like chondrocytes,and the extra cellular matrix also increased obviousl~.Furthmore,with the culture time extension,this change were more evident.HE staining showed that cells filled all the inter-connected pores in the constructs.And more cells were observed in the outer layer of the constructs.ECM(extraeellular matrix)Was strongly positive by Aleian blue,Safrine O staining and type Ⅱ collage immunohistechemical staining.DMB binding assay revealed that the induced BMSCs GAG secretion(1306.7±192.3)wag significantly higher than BMSCs(205.0±26.2)(P<0.001),but it was significantly lower than passage 2 ehondrocytes(1969.2±235.3)(P<0.001).Saltine O and type Ⅱ collage immunohistochemical staining were positive in constructs implanted subcutaneously.Conclusion Tissue engineered cartilage could be obtained using BMSCs and photografting modified PHBV,but there are still gaps physiologically between the constructs and the nature cartilage.
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OBJECTIVE@#To study application of sternocleidomastoid myoperiosteal flap to reconstruction of the defect of cervical trachea.@*METHOD@#Thirteen hospitalized patients with malignant neoplasm invading cervical trachea and primary tracheal carcinoma were analyzed retrospectively, the sternocleidomastoid myoperiosteal flap was applied to repair the defect of tracheal wall after resection of the neoplasm.@*RESULT@#Among 10 tracheotomy patients, 9 cases were decannulated from 1 month to 5 months. One case needs cannulation because of tracheal stenosis. Three patients with no tracheotomy had no dyspnea after operation.@*CONCLUSION@#The sternocleidomastoid myoperiosteal flap is an ideal transplant for cervical tracheal reconstruction.
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Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Músculos Peitorais , Transplante , Periósteo , Transplante , Procedimentos de Cirurgia Plástica , Métodos , Estudos Retrospectivos , Retalhos Cirúrgicos , Traqueia , Patologia , Cirurgia GeralRESUMO
BACKGROUND: At present, there are very big differences in structure,material character and biological property between artificial intervertebral disc (AID) and normal physiological intervertebral disc.OBJECTIVE: Three-dimensional finite element method was used to observe and analysis the stress conduction of artificial lumbar intervertebral disc in lumbar motion segment.DESIGN: Single sample observation was designed.SETTING: Department of Orthopaedics, Third Affiliated Hospital, Sun Yat-sen University; Department of Orthopaedics, Second Affiliated Hospital, Sun Yat-sen University; Laboratory of Mechanics, Southern Medical UniversityPARTICIPANTS: It was to employ a vertebral sample without any spinal disorder of a healthy male died due to accidence and a finite element model of AID implantation in vertebral motion segment established with SB Charite Ⅲ AID.METHODS: According to industrial design chart of AID, finite element software MSC.MARK was utilized to establish three-dimensional model of artificial lumbar intervertebral disc. The corpus sample of motion segment of healthy lumbar vertebrae was collected and scanned with spiral CT machine and imaging documents were input in computer to preserve.Geometric model of L4-5 segment was established in three-dimensional coordinate system in ASC.MARK software. The intervertebral disc in L4-5 motion segment model was replaced by AID. It was to ensure the fixation of lower terminal lamina of L5 in the model. 4 Nm moment of force was exerted in anterior flexion, posterior extension, lateral bending and torsion on the sample successively. Finally, force of internodes representing AID was calculated and stress distribution was recorded.MAIN OUTCOME MEASURES: To observe stress distribution of anterior flexion, posterior extension, compression, lateral bending and rotation of AID.RESULTS: Finite element model of artificial lumbar intervertebral disc implanted lumbar motion segment that is in conformity with clinical practice was established. Stress distribution of AID was characterized as:er lamina was the maximum and that in the lower inclined part of slide of slide core and cover lamina was two or three times as same as that of sion, the stress in the center of slide core and cover lamina was the maximum.CONCLUSION: The finite element model of artificial lumbar intervertebral disc implanted lumbar motion segment established is in conformity with the structural character of practical artificial intervertebral disc in morphology, size and motion property, based on which, it is feasible to carry on the experiment on stress distribution of artificial intervertebral disc.
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AIM:To evaluate the biocompatibility between copolymers of poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) and bone marrow stem cells (BMSCs). METHODS: Canine BMSCs were isolated and cultured. The cells in passage 3-4 were seeded onto the PHBV films and three-dimensional foams. The seeded cells were observed under inverted microscope for morphology and cell attachment onto the PHBV films at 1, 2 or 3 weeks after seeding. With 4% paraformaldehyde formalin and staining, the protein content in seeded cells was determined by bicinchoninic acid assay (BCA). The content of DNA was quantified using the Hoechst 33258 assay. RESULTS: Observation under inverted microscope showed that the PHBV fabric was fairly thickness, lucency is weak. Unser contrast phase microscope, PHBV fabric was uneasy to be observed. Most cells attached onto the PHBV films 2 h after seeding, and extended well and acquired a spindle fibrecyte-like morphology 3 d later. Moreover, on the three-dimensional foams, the seeded cells lay in micropores and grew tri-dimensionally. The conjunction of cells appeared about 1 week, and extended at 3 weeks, with a large amount of extracellular matrix around cells. The content of DNA and protein has no significant difference with control group. CONCLUSION: As a kind of tissue engineering material for BMSCs seeding, PHBV has an excellent biocompatibility.
