| ObjectIn June 2000, when I was at the age of 61-year-old, as a retired civil servant, I yearned to a comfortable retirement life. Unexpectedly, I suffered from palpitations in the second year and diagnosed with coronary heart disease and treated with heart bypass surgery in Chang gung memorial hospital in Taiwan. After a long-term use of Losartan potassium, division of attack prevention drug isosorbide-5-mononitrate cr, aspirin as a drug to prevent myocardial infarction and blood embolism syndrome, my cardiac function recovered gradually, but my kidney function had been declined year by year accompanied with hyperuricemia and gout, ureteral calculi, diabetes and other symptoms. Considering my father a TCM doctor, I started to recuperate with Chinese traditional medicine without side effects. In September 2010 I was admitted to Guangzhou University of Traditional Chinese Medicine to study TCM. In September 2013, I had got gout attack and was treated with experimental formula of my classmate Wu. I recovered with a bit relief of pain. Considered with the domestic economic prosperity, improved living standard and food culture by the impact of the western-style fast food culture, more and more people suffer from hyperuricemia and gout because of absorbance of substances of high purine, high protein etc and the onset age declined. Therefore, I proposed the study of "experimental and clinical research of Wuling Gout Pill in Hyperuricemia and gout". In the yellow emperor’s Canon, gout belonged to evil wind, paralysis in Jin and Yuan periods, the name, symptoms, differentiation types and formula of this disease were unified. TCM has advantages in the treatment of this disease. To verify its effectiveness and carry forward the traditional Chinese medicine and science, experiments.specially usage of concentrated TCM agent, and clinical trials of Wuling Gout Pill had been carried out.Method1. Theory research:To arrange systematically of pathogenesis, syndrome differentiation and treatment in evil wind.paralysis and gout disease according to the the ancient TCM literature.2. Animal experiments:(1) acute gouty arthritis model method:we took 60 male Wistar rats weighed 200+20 g, random divided into six groups:blank group, model group and the positive group (0.39 mg/ml Votaren). Rats in Chinese medicine high, medium and low dose group were given Traditional Chinese medicined 3 days prior to the modle having been built. Rats in positive group were given medicine 2 days prior to the modle having been built. Except the blank group, all the other groups were given 0.2 ml uric acid sodium solution injected into the stamping articular cavity, the formation of acute gouty arthritis model. (2) cell experiments:(1) building methods:to explore the method of grope for uric acid salt concentration of building and building time: to culture primary synovial cell of rats, three concentrations of uric acid salt were put into the medium (7001000,1,400 uM) and detect the expression of VCAM 1, synovial cell. (2) "Wuling Gout pill" medicated serum preparation: to explore the drug concentration and dosage of time of Chinese traditional medicine an to detecte of level of VCAM 1, PCR and WB. To build culture rat primary synovial cells and put them into different groups.3. Clinical trial:(1) 18 patients were outpatients of the first affiliated hospital of guangzhou university of TCM from October 2015 to January 2016 and 12 patients came from shenzhen wu Qiu Bao western medicine internal medicine clinic. All the paitents were put into control group and TCM group randomly. Clinic symptoms of the two groups were compared before and after taking Chinese and western medicine. All the patients were given diet therapy and patients in TCM group were given Wuling Gout pill 28 grams per day, points 2 times/or 14 bid with hot water about 100 ml of solution every time an hour before meals, which had the effect of promoting blood circulation to remove blood stasis, swelling and pain when gout attack. Patients in control groups were given western medicine treatment such as benzene bromide Malone, allopurinol tablets, sodium bicarbonate and colchicine treatment.Content:1. Animal experiment:Effect of Wulingtong gout pills on rat model of acute gouty arthritis:(1) building time:on May 5,2015-on May 19,2015. (2) building method: 60 male Wistar rats (200,20 g) were divided into six groups and each group had five rats. ① Rats in blank group and model group were given with physiological saline lavage, one time a day, for 5 days. ② Rats in Voltaren group were given with diclofenac sodium (suspension liquid lavage (0.39 mg/ml), 1 time a day, for 5 days.③ Rats in high dose TCM group were fed with thickening high concentrations of 28 g solution of Wulingtongfeng 1 time a day, for 5 days. ④ Rats in medium dose TCM group were given with medium concentrations of 20 g solution of Wulingtongfeng 1 time a day, for 5 days.⑤). Rats in low dose TCM group were given with medium concentrations of 12 g solution of Wulingtongfeng 1 time a day, for 5 days. Except the rats of the blank group, rats the rest 5 groups, were made mould, injection of uric acid sodium solution crystallization, induced acute joint model. Rats in blank group were given with the same volume of physiological saline. At the fifth day,24 h after being filled the stomach out, all rats were put to death.2. Clinical experiment:Caused by high uric acid hematic disease, acute gouty arthritis is prodrome of gout with sudden onset. The starting symptom is the foot of the great toe in most cases. The second affected areas respectively the ankle, knee, wrist, with joints, elbows, etc. Usually there are a variety of incentives, such as meal, eat high purine food, alcohol, fatigue and cools down, infections, etc. The period of onset could continuous from a couple of hours to days or weeks. Remission period may be months or even a lifetime. Caused by the repeated gout attacks, chronic course of the disease could exist. The theraputic methods conclude declining the blood uric acid, alleviating the symptoms of acute phase, reducing the recurrence of acute. The treatments in western medicine mainly focus on relieving the symptoms and anti-inflammatory analgesic detumescence quickly with big side effectswhile Traditional Chinese medicine treatment of gout has significant advantage lacking of side effects.3 Diagnosis standard of hyperuricemia:according to the domestic data blood uric acid of normal male in 150-380 umol/L (2.46.4 mg/d1), uric acid of premenopausal women is 100-300 mu mol/L (1.65.2 mg/dl), and uric acid levels close to normal postmenopausal women men. With any reasons, the blood uric acid caused no matter by increased production or by less discharge, the cases in which the higher concentration of blood uric acid blood uric acid concentrations for men and postmenopausal women are more than 380 mu mol/L (more than 6.4 mg/dl), premenopausal women is more than 300 mu mol/L (5.2 mg/dl) can be diagnosed with high blood uric acid.4 Classification criteria of acute gouty arthritis:according, to the 1977 American rheumatism association (ACR) set of the acute gout arthritis diagnosis reference standard,5 Diagnostic criteria of chronic kidney disease (CKD):on chronic renal insufficiency in our country is divided into five periods:the stage 1 normal (for kidney disease), stage 2 compensatory stage, stage 3 decompensation stage, stage 4 renal failure,5 uremia.6 TCM diagnostic criteria:refer to 1994 issued by the state administration of traditional Chinese medicine, Chinese medicine of the People’s Republic of China industry standard, the standard of diagnosis of disease and curative effect of traditional Chinese medicine ZY/T001.1-94 medical gout syndrome diagnosis of TCM curative standard.7 Clinical trial ethics standard, etc.Results1. Animal experimental results:(1) mechanism research:The results showed that:Votaren group and high dose group of Chinese medicine were related to the decreasing of joint dysfunction index. Joint inflammation index of Chinese traditional medicine group was lower than that of otaren group with obvious effect in 24h and 72h. After 24 h and 72 h, joint high swelling rates of Chinese traditional medicine group of were lower than Votaren group, and also lower than that of model group. Joint synovial tissue pathological changes in morphology, we found that Chinese traditional medicine could reduce the inflammatory cells infiltration, hyperplasia of phagocytes and tissue disorder, etc. Finally, by Elisa and immunohistochemical experiments found that the expression of NF K appa B p65 predominate, VCAM1 and IL-1 beta in Chinese traditional medicine group decreased in various degrees associated with dose effect, namely the higher dose group, Chinese traditional medicine has the higher efficiency. That traditional Chinese medicine for the treatment of acute gouty arthritis has a certain effect, and we suspect that the nuclear transcription factor NF K appa B predominate signaling pathways play an important role.(2) Mechanism research of Wuling Gout pill:Through the cell experiment: first of all, by building the cells of gouty arthritis model and through the qRT-PCR test, we detected the expression of ICAM-I, to grope for the uric acid salt concentration. We found that 24h after model building is the best time.1000 mu M Secondly was the best Chinese medicine concentration about the production of large Numbers of inflammatory factor for 24h best. Through qRT-PCR test, the expression of ICAM-I was detect. It was found that with the increase of concentration, ICAM-I dropped associated with dose effect. Therefore, it was the best effect in high dose group. In order to verify NF kappa B predominate signaling pathways play a role, we speculated that NF kappa Bp65 predominate siRNA interference cut its expression (through the qRT-PCR and WB experimental screening effective siRNA). The WB and Elisa experiments results showed that the NF kappa Bp65 predominate after cut, NF kappa B p65, VCAM1 predominate and IL-1 beta expression, and the effect is almost the same with Chinese traditional medicine group, thus speculated that traditional Chinese medicine (TCM) could weaken the expression of NF kappa B predominate, affect VCAM1 and IL-13 expression and play an important role.2. Clinical research results:(1) with reference to the clinical disease diagnosis according to the definition of the heal better relative content in the 2002 version, clinical practice Hyperuricemiaformulate curative standard were formed.40% patients were effective markly and 33.3% were effective and 26.7% invalid. The total effective rate was 73.3%. (2) Comparion with renal function and blood uric acid before and after the treatment, blood uric acid in TCM group was 528.8+69.9 mmol/L, before treatment and after treatment uric acid reduced to 435.6+78.0 mmol/L on average with statistical significance (p< 0.05). It showed Wuling Gout pill could drop uric acid with obvious curative effect. (3)The glomerular filtration rate (eGFR) and eGFR was 68.93+17.22 ml/min on average before treatment, the average eGFR was 77.09+ 20.75 ml/min after treatment. It seemed to be improved after treatment, but after a statistical test found that there was no statistical difference between the fork (p> 0.05), which showed that Wuling Gout pill be harmless to renal function by uric acid treatment.ConelisionThe statistical analysis results of the animal experiment, cell experiments and clinical research and are the following:(1) module swelling rate of Wuling Gout medium dose group was lower than that of the high dose group. It suggested that high dose group was effective than low dose group and control group. (2) Wuling Gout had certain anti-inflammatory effects in animals, and the gouty joint swelling inhibition effect significantly. (3) In the cell experiment, Wuling Gout could reduce the local lesion of gouty arthritis of rats inflammatory white blood cells, the sliding mode infiltration and hyperplasia ccording to the experiment (4) Wuling Gout for gouty arthritis induced by uric acid salt, had a certain therapeutic effect, can reduce inflammatory reaction, by the whole experiment proved that is a nuclear factor transcription factor NFk Bp65 signaling pathways which played a role of inhibition. (5) According to three gout capsule on acute gouty arthritis impact experiment, it proved that it has anti-inflammatory effects, and also discusses some of its mechanism, so its experimental results, the theoretical basis of clinical provides strong. Proved by traditional Chinese medicine with severely, persistent effect and less side effects, in addition to the primary symptom gout treatment, to its accompanying disease is comprehensive. (7) Clinical research conclusion:the statistical analysis showed that:① The remarkbly effective rate of was 40%, effective rate was 33.3% and invalid rate was 26.7%. Total effective rate was 73.3%. ② Wuling Gout Pill could decreased blood uric with statistic signficance. It showed Wuling Gout pill is effective to drop uric acid obviously. ③Comparied with before and after the treatment, Glomerular filtration rate was improved without statistical significant. It showed that Wuling Gout pill is harmless to renal function. |
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