| 1. ObjectiveThe morbidity of osteoarthritis is yearly increasing due to aged patients, social stress and rapid rhyme of modern life. Osteoarthritis of the knee is a common degenerative joint disease in middle and aged people. The basic pathological characteristic of it is that it involves degenerative or vanishing tissue in the arthrodial cartilage, osteophytes in the joint margin and secondary changes in subcondral bone. The whole process involves the whole joint, including subcondral bone, ligament, joint, synovium surrounding muscles of the joint. It results in aching in the joint and dysfunction. Owing to the high morbidity, complicated condition, long history, affecting on life and work seriously, approaching osteoarthritis of the knee becomes important part by clinic workers.In present research, the method of treatment in osteoarthritis of the knee is symptomatic treatment, such as pharmacological, physical and surgical therapies, and the therapeutic effect is so limited that it is difficult to control accurately. It is also difficult to apply to the clinic , since the approach on chondrocyte grafting, cartilage grafting, tissue engineering grafting and genetic therapy is lack of the source and quantity. However, MSCs, which is easy to obtained, causes minor trauma, has strong ability of proliferation and good differentiation in vitro, as well as have no immunological rejection because of the tissue induced by auto stem cells. It is popular to be applied by the medical research workers. It is proved that MSCs are precursor cells of many kinds of mesenchyal cells, such as osteoblast and chondroblast.In the view point of TCM, osteoarthritis of the knee belongs to the category of "paralysis of bone" . Plain Question says: "patients can not rise their limbs, feel marrow aching pain if the bone is involved because of coldness, this is called paralysis of bone" ; ZhangShiYiTong says: "the knee is the collectivity of the tendons; knee pain results from deficiency of the liver and kidney" . The theory of TCM considersthat the kidney dominates bone and the liver controls tendons. In the middle age, the essential of the kidney and the liver declines, the bone is poor of nourishment and give rise to pain. Also because of the exogenous factors of wind, cold, damp invading and stagnating in the joint, or because of traumatic injuries, the blood circulation of the bone is stagnating, and the pain can be manifested by the stagnation. The key method of treatment in TCM lies in improving fundamental and treating incidatal, such as tonifying the liver and kidney, removing the wind and cold, removing the damp and relieving pain. Chinese medicine , which has minor toxic and side effect and has protective function to the body cells, are applied to clinic . Because of low repeatability, the limiting level as a whole in the experimental study, many prescriptions in differential symptoms, few mechanisms is explored. It is difficult to realize industrialization.According to the mechanism of modern study, the guidance with the theory of TCM and the long period of clinical experiments, we select Chinese medicine as a peroral dosage form of complex prescription, which can vital qi and tonify the kidney, dispell wind and remove obstruction in the meridians, relieve pain. Serum pharmacology and clinical experiment are made, the criterion of the diagnose in the patients with osteoarthritis of the knee is consummated , the criterion of the therapeutic effect is also systematize. This study is aimed at identifying the function of Chinese medicine, exploring the mechanism of it' s treatment, investigating the affect on the proliferation of MSCs in vitro with the serum containing Chinese medicine, which lays foundation in the development of Chinese medicine and Chinese medicine combined MSCs in the treatment of osteoarthritis of the knee.2. Method and ContentClinical research: Referenced by "the Criterion of the Diagnosis and the Therapeutic Effect" published by State Administration of Traditional Chinese Medicine , osteoarthritis of the knee .which is called bone paralysis in TCM , classified as a type of deficiency of kidney and marrow, manifested by pain of the knee joint, loss of movement,. acid and soft in the lumbar and the knee, dizzy, tinnitus, pink tongue, thinness and whiteness moss, micro string impulse. According to the criterion of thediagnosis, internalize and exclusive criterion is made. The groups are classified strictly by random and double blind method , while the criterion of therapeutic effect is consummated. Clinical research is made by the following aspects: (1) the observation of the clinical symptoms and signs(the assessment of function), (2) the observation of the imageology (X-ray), (3) the observation of the peripheral blood , such as MMP-3, TNF-a, IL-1, hyaluronic acid (HA) , SOD and LPO, (4) the observation of the routine examination of blood, urine and stool, the hepatic and renal function .Meanwhile, combined with the above data, and with the view of the concept of the wholism in TCM, the mechanism of the pathological in osteoarthritis of the knee and the function of Chinese medicine is explored.Labolatory research: (1) With the identification applied by the cytomorph and phenotypic expression(HEstain:the cytomorph is fibroblast-like, the identification of phenotype in immunocytochemical method is CD34-, CD44+, Ckit+), the system of the culture of rat marrow mesenchyal stem cells is established. (2) With the serum containing Guanjiekang is collected, the growth curve of MSCs with the 10% serum is drawn to investigate the function of the proliferation of MSCs in vitro. Statistical method: numeration data is statistic with X2-test, measurement data is statistic with t-test.3. ResultsClinical research:(1) The observation of the clinical symptoms and signs: The ache of movement after treatment both have improved significantly (fKO. 05)in therapy group and control group. Therapy group have improved significantly after 1 month of treatment. Compared with itself, there is significant difference, And there is no gastrointestinal tract adverse effect. The range of movement, stability, varus and valgas are improved after treatment, However, there is no significant difference(f>0. 05). Flexion deformity have improved significantly after 3 month of treatment (K0. 05). The straighten delay, the distance of walk and climbing steps have improved significantly. Compared with itself, there is significant difference.(2) The observation of imageology (X-ray) : With Kellgren and Lawrence grading criterion:1 and 2 grade are I stage, 3 and 4 grade are IIstage. There is no change and no significant difference in X-ray after treatment.
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