Global Analyses Of Gene Expression In Primary Osteoarthritis Cartilage Of The Knee Joint Of Humans

Osteoarthritis of the knee(KOA) with age is a significant increase in the incidence of degenerative joint disease,joint pain and the elderly are the main reasons for disability. According to the World Health Organization statistics,osteoarthritis prevalence rate in women in the fourth.Increases with age,the prevalence rate rising rapidly,more than 65-year-old more than 50%of people have osteoarthritis of the performance of X-ray film.It's main change is the surface of articular cartilage degeneration and secondary hyperplasia of the bone.The main performance and activities of joint pain is not flexible, X-ray manifestations of joint space narrowing,subchondral bone density,bone fracture, there is sclerosis and cystic change.Joint edge lip hyperplasia.The latter end of the deformation of bone,articular surface uneven.Off articular cartilage,bone fragments into the joints,the formation of intra-articular loose bodies.KOA etiology can be divided into primary and secondary KOA,primary KOA, refers to the current method of examination of all the bone unable to cause arthritis, osteoarthritis is usually referred to fall into this category;secondary KOA is the other cause or basis of disease,-induced lesions,such as trauma,rheumatoid arthritis, neurological and endocrine diseases.This type of lesions of osteoarthritis more limited, not associated with Heberden nodules.Repeated strain of the crowd so that the joints suffering from osteoarthritis is high-risk groups,such as foundry workers,miners and bus drivers.But for long-distance train are not suffering from this disease in high-risk groups.Obesity is a major factor in osteoarthritis,but the evidence is not sufficient.In accordance with the KOA site can be divided into focal and generalized KOA,in accordance with whether there are clinical symptoms KOA can be divided into asymptomatic and symptomatic KOA.Encountered in clinical work KOA are symptomatic KOA.Mainly against articular cartilage,bone and synovial tissues,leading to joint pain,deformity and dysfunction,thus affecting the activities of patients.Under normal circumstances,very little friction between the joints will not cause wear and tear, unless the excessive use of or injury.Caused by osteoarthritis is the most likely cause of the abnormal synthesis of cartilage components,such as collagen(a tough,fibrous connective tissue protein) and mucin(a substance produced flexible cartilage) anomalies In addition,the growth of cartilage while strong,but thin,its surface is prone to rupture. Joints over the edge of the bone growth,can see and touch the formation of the mass (known as osteophyte).Osteopbyte caused by uneven articular surface,interfere with the normal function of the joints,causing pain.Articular cartilage from disease onset,affects the entire joint structure,including the subchondral bone,ligaments,synovial membrane,joint capsule and muscles,the ultimate result of all the loss of articular cartilage leading to joint deformity and loss of function of a disease.Articular cartilage is an important factor in the pathogenesis of the articular cartilage is a 1-2mm thickness of collagen fibers,glycoproteins,hyaluronic acid ester gathered together,when the hydration on a mat-like role in the absorption and dispersion the burden of weight-bearing and mechanical strength.In physiological conditions,relying on the articular cartilage and thermal contraction of the surrounding subchondral bone and to complete the above tasks.In addition to stimulate muscle contraction joint activities,at the same time plays the role of rubber band-like, absorbing a great deal of momentum from the protection of the joints.When an accident occurs(eg,wrestling),because this sudden shock the muscles can not react in time of emergence of protective weight-bearing joints increased virulence may be caused by joint injury,so this sudden burst of muscle vibration protection can not be there in time to reflect heavier load of the joints can be caused by joint injury.In addition,muscle aging,peripheral neuropathy,the muscle function of energy absorption also decreased significantly.Weight-bearing cartilage for the assistance of another factor is the network showing the distribution of subchondral bone quality,its texture,although more cartilage than bone cortex should be soft,it has a high degree of flexibility will help to bear the pressure.Osteoarthritis can be seen more new in the following two situations: First,articular cartilage,subchondral cortex and abnormal muscle around the joints, such as age-related degeneration,osteoporosis,inflammation,metabolic diseases,such as:The second is the articular cartilage of the joints under the bone,the muscles around the joints,although subject to normal due to excessive pressure,such as obesity,trauma, etc.Chinese medicine treatment of knee osteoarthritis has a wealth of clinical experience and unique strengths.With the traditional Chinese medicine treatment of knee osteoarthritis Experimental study on the deepening of Chinese medicine for the treatment of knee arthritis awareness of the role of the mechanism,the organization has changed from simple morphological analysis of the development of immunohistochemistry,molecular biology,such as multi-disciplinary,multi-level, multi-session awareness.In recent years,a variety of experimental study found that traditional Chinese medicine for knee osteoarthritis mainly reflected the impact of knee osteoarthritis in the organization of morphology,cell factor,bone pressure,oxygen free radicals and so on.KOA diagnosis usually relies on clinical manifestations and radiological examination.Its diagnosis is based on simple.1.Symptoms:â‘ pain:pain during exercise or by mechanical tendon,ligament caused by joint disease;rest for the pain caused by inflammation.Night pain prompted increased intraosseous pressure,in order to be in a serious condition and poor prognosis indications.â‘¡Rigid:in the joint after inactivity,it is generally not a long duration.â‘¢Other symptoms such as joint swelling,deformity and joint ring,such as shells.2.Signs:the sound of joint,bone hypertrophy,various degrees of synovitis can be caused by tenderness,abnormal tendon reflexes,such as myasthenia gravis.Serious cases of dysfunction or disability can be seen. 3.To observe the patient walking,bending and the ability to carry out daily activities.4.Imaging examination:including X-ray,CT and MRI examination.5.Other tests:blood test,examination and synovial fluid,such as arthroscopy.However,in the early and could not KOA radiological examination revealed the changes in articular cartilage,as can be found in the radiographic changes of cartilage prior to a certain cartilage metabolism and ultrastructure in the elements have taken place in very significant changes.KOA in primary clinical research and experimental studies although many efforts have been made,the study shows that articular cartilage and synovium in the pathogenesis of primary KOA play a leading role,but its pathogenesis is still not very clear,the current operations and take more symptomatic therapy.ObjectiveArticular cartilage in the study subjects for the purpose of the adoption of whole-genome gene expression,research and all the disease-related differences in gene expression,thus further explore differentially expressed genes in the cartilage and the relevance of this disease,for treatment of this disease new target and new methods.At the same time,some differences in gene-hole complex real-time quantitative PCR,to further verify the differences in access to primary KOA gene in pathological significance in the process.MethodsMaterialAccording to addition and exclusion standards,12 patients with OA were performed,and OA cartilage of the knee joint of humans was obtained at the time of arthroscopic surgery and total knee replacement.According to Kellgren and Laurence radiology classification standards 4 patients became a group,then mild(R2) and moderate(R3) and severe late-stage OA groups(R4) appeared.In addition,4 normal cartilage of the knee joint samples became RBl group.Methods1.All the 16 samples were performed the genome-wide analysis of chips,each chip has 40,000 point samples detected 16 cases of co-owned by the gene expression.2.According to the foregoing groups,the genes within each group were corresponding calculated the mean,each group was considered as a entirety (R2,R3,R4,RBl four groups),comparing among the four groups.3.RNA sample preparation(1) two-dimensional separation(2) PNA praecipitatum(3) RNA emundation(4) again dissolve RNA praecipitatum(5) RNA mass detection4.aRNA sample symbol and synthesis(1) cDNA synthesis(2) second-strand of cDNA synthesis(3) aRNA synthesis(4) aRNA depuration(5) aa-UTP aRNA indirect labelling(6) quality control5.Chip prehybridization6.Hybridisation7.Chip emundation8.Result scanning,data reading9.Synthetical cDNA were used to real time quantitative polymerase chain reaction10.Performing real time quantitative polymerase chain reaction Results1.In this experiment,among the 41000 genes,many differential expression genes were identified,and 70 genes showed statistical significance,of which: 46 genes were up-regulation 3-fold,thereinto 28 known genes and 18 unknown genes,in known genes,25 genes were up-regulation 4-fold,15 genes were up-regulation 5-fold,11 genes were up-regulation 6-fold, 8 genes were up-regulation 7-fold,3 genes were up-regulation 8-fold;24 genes were down-regulation 3-fold,thereinto 16 known genes and 8 unknown genes,in known genes,11 genes were down-regulation 4-fold,8 genes were down-regulation 5-fold,6 genes were down-regulation 6-fold, 6 genes were down-regulation 7-fold,2 genes were down-regulation 8-fold.2.More than up-regulation 3-fold of difference genes include:participating gene of cartilage's catabolism and gene with immunologic function.3.More than down-regulation 3-fold of difference genes include: participating gene of cartilage's anabolism and gene with information transfer function.4.In 70 differentially expressed genes in any differences in the selected 10 genes Were optional selected to perform real time quantitative polymerase chain reaction,through three times repetitive experiments,we miraculously observed the same results of the experiments.Through the statistical treatment, P values less than 0.01,There are significant differences.5,During R2 and R3,R3 and R4,R2 and R4 comparison,up-regulation or down-regulation than 3-fold difference in gene expression in the course of osteoarthritis lesions in no significant difference has nothing to do with disease severity.Conclusion1.KOA's occurance is relate to differentially expressed genes of articular cartilage2.KOA's pathological changes is irrelevance with differentially expressed genes of articular cartilage...