| Part Ⅰ:The experimental study:Jieguyao Pill Inhibites The Function Of Osteoclast-Like-Cell Differentiated From Spleen Multipotent Stem Cell:Bone Resorption.Objective:By establishing vitro culture system of osteoclast, we try to explore Jieguyao Pill’s therapy of inhibiting the differentiation and bone resorption effects of osteoclasts.Methods:20Wistar rats were randomly divided into4groups, the two experimental groups were respectively administered Jieguyao pills(Group A) and total flavonoids of drynaria (Group B), control groups to take Alendronate solution(Group C), while the blank control group were given intragastric administration of distilled water(Group D),the rats were administered once every day.14days later the blood of each rat was drawn of from the heart, by high-speed centrifugal the supernatant fluid was just blood serum, which should be stocked cold standby.Then spleens of fetal rats were cut off the stomach under aseptic condition, to go through grind,filtration and centrifugal, the supernatant was discarded,and spleen pluripotent stem cell suspension were diluted with a-MEM medium which was cultivated for72hours.To go through digestion and high speed centrifuge again we discarded the supernatant and prepared cell-suspension of5×105/ml constant volume with culture solution, which contained the sM-CSF and RANKL concentration.8culture plates(6holes) was divided into4groups,and24microscopic slides and24bone fragments were respectively put into the holes in advance. Then we joined0.2ml cell-suspension into every hole, then four kinds of serum was added into the corresponding set of holes, under a microscope we watched the condition of cell growth everyday.10days later, the slides were taken out of the holes and stained with Tartrate-resistant acid phosphatase and the number of positive cells was counted; the bone fragments were stained by1%toluidine blue and the bone resorption lacuna showed,then we counted the number of lacuna under the electron microscope. Results are indicated by mean±standard deviation(x±s), SPSS17.0software was applied to make statistical analysis using t test.Results:Each set of cells in the holes were in good condition,7-9days after culture irregular cells with several nuclears occupied much of the vision, and after TRAP staining these red cells showed characteristics of osteoclast,on the surface of bone fragment a number of blue lacuna also appeared.Both numbers of Group A and Group B counted less than Group D which has statistics significance(respectively P<0.01and P<0.05), but the comparison between Group B and Group A displayed obvious difference(t test P<0.01); however, the TRAP dyeing positive cells’number and the bone absorption depression of Group A were less than Group C, and statistical analysis also display significant difference(P<0.05).The self-made Chinese medicine of our hospital Jieguyao pills and total flavonoids of drynaria can suppress osteoclast-like-cells’mature and its function of bone resorption,which differentiated from spleen multipotent stem cells.But the Jieguyao pill, as set prescription, the effect was much better than total flavonoids of drynaria, which some kind reflected the rationality of Chinese medicine prescription compatibility theory; However, Jieguyao pills,as adrug to promote fracture healing, its effectiveness of contra-osteoporosis was a little weaker than Alendronate, the most popular contra-osteoporosis drug. Part Ⅱ The Clinical Study:Applying Jieguyao Pill TO Treat Primary OsteoporosisObjective:We try to study the self-made prescription Jieguyao pill’s effect in treatment of primary osteoporosis,explore the feasibility and effectiveness of jieguyao pill’s clinical treatment on primary osteoporosis, and provide theoretical and practical basis.for clinical medication of drugs.:Methods:We collected76patients with primary osteoporosis.who accepted therapy in Wendeng City Orthopedics hospital between March2010and January2011wendeng.By random numbers we randomly divided the patients into treatment and control groups with the38cases respectively. Two groups of patients would be administered Jieguyao pills(6g) and. Alendronate(lOmg) respectively at9AM.Both groups were continuously admistered for3months. During the period we detected the liver and kidney functions to ensure the safety of the medication. Respectively before and after the treatment Wards triangle’s bone mineral density was tested; bone metabolism markers, including serum calcium, phosphorus, alkaline phosphatase and urinary calcium/urinary creatinine were detected as well. By osteoporosis treatment efficacy criteria we distinguished the effects of two groups, and based on evaluation standards we ensured how many TCM syndromes were improved.Results:35cases of treatment group and35cases of the control group entered the result analysis at last. The TCM symptoms of Treatment group were improved much better than Control group’s,which showed obvious difference, and in bone metabolism index comparison aspects, there were no obvious changes in blood calcium,serum phosphorus and the alkaline phosphate, only the anhydride ratio urinary calcium/urinary creatinine wre obviously reduced,bone density value of both groups were improved as well.But the comparison between two groups display no obvious differences, even the control group showed more treatment than the treatment group.Therefore, the final total efficiency of the Treatment group was62.6percent, while the control group was64.7percent. Although the comparison of the two groups did not show significant differences, but the control group was slightly better than treatment group.Conclusion:Applying TCM basic theory to conduct Jieguyao pill’s usement in treatment of primary osteoporosis, we can significantly improve patients’discomfort symptoms, but in terms of promotion of calcareous deposits and increasing bone density,Jieguyao pill may not have obvious advantages compared with Alendronate.If we can reasonably combine Jieguyao pills with Western medicines such as Alendronate,the effectiveness can serve to complement each other. |
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