| Objective:To investigate the effect of Jiegufang on ON in mRNA gene expression in the process of fracture healing though the animal' s experiment and clinical research, and to explore the mechanism of Jiegufang in enhancing fracture healing.Methods:The animal' s experiment: forty female SD rats were produced by operation a closed-fracture experimental model in the right middle femoral, were divided equally into two groups (treatment and control)at random, rats in treatment group were taken with Jiegufang liquid by 10.4mL per kilogram, while in control group were taken with the same volume isotonic Na chloride. Animals in two groups were killed respectively after operation of 2w and 4w. Took out the callus in sterile circumstance and kept them frozen in Liquid Nitrogen. firstly, to observe the variance and differentiation of callus disposed with different measurements at different phases by chemistry staining, which included routine paraffin section, and HE taining. Secondly, The expression of the mRNA for ON was investigated by in situ hybridization technique during the healing process of experimental fracture.The clinical research: 48 radius fracture patients (2006.9～2007.2) were randomly divided into treatment group and control group. Treatment group were given Jiegufang and shangkejiegupian, control group were given shangke jiegupian. Both groups take medicines for one month. After one month treatment, we observe the count-scores of clinical symptom and Callus.Results:The animal' s experiment: (1) Model: closed fracture models were made good, among them middle-fracture about 87.5%, transverse fractures 92.5%. (2) callus and HE staining: on the 14th day, both the drug treatment group and the blank group were all masses of callus from fracture site. a great deal of new bony trabecula and chondral tissue connected with each other were found at the fracture ends in group B, where osteoblasts and chondroblasts were active; hematoma disappeared thoroughly in group A, fibrous tissue proliferation were active, a great of osteoblasts came into being, and so were some chondral tissue, on the 28th day, In group B, inner-callus connected with outer-callus mostly at the fracture ends, the shape of outer-callus looked like compact bone, much of bony trabecula were well distributed in size and allayed in the same direction, and possessed good refraction under microscope, as to group A, bony trabecula allayed in disorder, and refracted unevenly, much of fibroblasts, chondroblasts, osteoblasts and osteoclasts were found in fracture site. (3) ISH: the expression of ON mRNA in the chrondrocytes within proliferative zone were more intensive, while chrondrocytes within hypertrophic zone and zone of calcifying cartilage showed absence expression. the intensive signal could be detected in the newly formed woven bone. The osteoblasts lining on the surface of lamella bone showed absence or only week expression, in the control group, the expression level of ON mRNA in 2w postoperation was apparently lower than that in 4w, which had a significant statistical difference (P＜0.05). in the treatment group, the expression level of ON mRNA in 4w postoperation was higher than that in 2w, which had no significant statistical difference(P＞0.05). As compared the treatment group with control group, the expression value of ON mRNA in the former was higher than that in the latter in 2w, but the result was on the contrary in 4w, which had the obvious difference (P＜0.05).The clinical research: The total effective rate in the two treatment groups was 95.7% and 86.4% at the aspect of symptom and signs. There was significant difference(P＜0.05). The contrast between pretherapy and posttherapy of the two groups all had significant difference (P＜0.05). At the aspect of fracture healing, the total effective rate of treatment group was 95.7%. The total effective rate of control group was 90.9%. they had no significant difference(P＞0.05).Conclusions:According to the experiment results, our understanding of the fracture healing pathomechanism and review of a series of research of Chinese herbal Jiegufang, yielded the following conclusions: (1) the standard closed-fracture models gained great success, which may be taken as a ideal animal model-made method. (2) Chinese herbal Jiegufang could obviously improve clinical effect of pain, swelling, petechia, local tender, percussion pain and callus, increase fracture healing course. (3) Chinese herbal Jiegufang could activate the competence of fibroblasts, osteoblasts and chondroblasts, and auxo osteoblasts proliferating, polarizing, mineralizing and ossifying, so it could accelerate fracture healing course, besides, Jiegufang might regulate the proliferation of chondrocytes. (4) The molecular mechanism of Jiegufang to enhance and increase fracture healing was that Jiegufang improved the expression level of ON mRNA and turned chrondrocytes repair into ossifying ane remodeling phase in advance.
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