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Objective:To establish an optimal extraction technology for gastrodine in Tianma Gouteng decoctions. Methods: An orthogonal design with extract yield and gastrodine concentration as the indices was carried out to observe the influence of the extraction time, water amount and extract times on the extraction result. Results:The extract times showed significant influence on the extraction percentage of gastrodine. The best extraction parameters were as follows:adding 12-fold water and decocting 3 times with 1. 5h for each time. Conclusion:The established extraction process is feasible, which can be used in the preparation of the effective part for Tianma Gouteng decoctions with gastrodine concentration as the index.
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BACKGROUND:Adipose-derived stem cel s and bone marrow mesenchymal stem cel s are used widely in cartilage tissue engineering, and there are many similarities in biological characteristics between two kinds of cel s. OBJECTIVE:To compare the chondrogenic potential of bone marrow mesenchymal stem cel s and adipose-derived stem cel s in vitro. METHODS:Adipose-derived stem cel s were isolated from the 3-month-old New Zealand white rabbits’ abdomen. Bilateral femurs of rabbits were obtained, and then the bone marrow mesenchymal stem cel s were separated with the adherence screening method. The growth curve of the passage 3 adipose-derived stem cel s and bone marrow mesenchymal stem cel s were drawn, and the doubling time of two kinds of cel s was compared. Then the passage 3 adipose-derived stem cel s and bone marrow mesenchymal stem cel s were treated with chondrogenic induction. After induced for 14 days, the adipose-derived stem cel s and bone marrow mesenchymal stem cel s were treated with toluidine blue staining and type Ⅱ immunohistochemistry staining respectively. RESULTS AND CONCLUSION:Primary bone marrow mesenchymal stem cel s showed aggregative growth, while the primary adipose-derived stem cel s were in single and scattered growth. The proliferation speed of adipose-derived stem cel s was faster than that of bone marrow mesenchymal stem cel s, while the doubling time of adipose-derived stem cel s was shorter than that of the bone marrow mesenchymal stem cel s. After chondrogenic induction for 14 days, both adipose-derived stem cel s and bone marrow mesenchymal stem cel s could express glycosaminoglycans and type Ⅱcol agen, and the expression level of type Ⅱ col agen in bone marrow mesenchymal stem cel s after chondrogenic induction was higher than that in the adipose-derived stem cel s. The in vitro proliferation of adipose-derived stem cel s and bone marrow mesenchymal stem cel s was rapid and stable, but the proliferative ability of adipose-derived stem cel s was faster than that of bone marrow mesenchymal stem cel s. When cultured in single layer, both adipose-derived stem cel s and bone marrow mesenchymal stem cel s could transform into chondrocytes under certain conditions, but bone marrow mesenchymal stem cel s seemed to be more potential than adipose-derived stem cel s.
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BACKGROUND: Now, dual-energy X-ray absorptiometry is international y recognized as gold standard for the diagnosis of osteoporosis, but the errors can be found in the measurement results due to the heterotopic ossification and bone hyperplasia exists in the measurement part. OBJECTIVE: To investigate the clinical significance of bone metabolic markers in the diagnosis and treatment of elderly patients with osteoporotic fractures, and to research its correlation with the changes of pathological histology and bone mineral density. METHODS: Four bone biochemical markers in 50 elderly patients with osteoporosic fractures were measured preoperatively. According to the results, 25 patients had significantly increased tartrate-resistant acid phosphatase 5b (considered as the increased tartrate-resistant acid phosphatase 5b group), and 25 patients had increased bone alkaline phosphatase (considered as the increased bone alkaline phosphatase group). During operation, the bone tissues of eight patients in each group were treated with hematoxylin-eosin staining and electron microscopy scanning in order to detect the pathological changes. After operation, the patients in the increased tartrate-resistant acid phosphatase 5b group received salmon calcitonin anti-osteoporosis treatment, and the patients in the increased bone alkaline phosphatase group received the anti-osteoporosis treatment of bone peptide injection. The bone mineral density and the four bone biochemical markers were detected again at 6 months after treatment. RESULTS AND CONCLUSION: There were no significant differences in the preoperative bone mineral density and four biomechanical markers between two groups (P > 0.05). The pathological examination results of bone tissue on the fracture site showed that the number of osteoblasts was reduced and the number of oeteoclasts was increased in the increased tartrate-resistant acid phosphatase 5b group; while in the increased bone alkaline phosphatase group, the pathological examination results showed the number of osteoblasts was reduced; the trabecular bone/bone area ratio was decreased in two groups, and there was a significant difference in the decrease degree between two groups (P < 0.05). The electron microscope scanning showed that the osteoclasts of two groups were more active than that of the normal group. The sloppy of trabecular bone in the increased tartrate-resistant acid phosphatase 5b group was more obvious than that in the increased bone alkaline phosphatase group, and the absorption vacuoles were increased. There were significant differences in the bone mineral density and four biomechanical markers between two groups before and after anti-osteoporosis treatment (P < 0.05). The detection of bone metabolic markers could help us to make it clearly that the main function of osteoblast reduce or osteoclast increase in bone tissue of patients, and guide us to use anti-osteoporosis drugs in target. Pathological histology examination can better reflect the condition of osteoblasts, osteoclasts and trabecular bone in bone tissue on the fracture site. Target application of anti-osteoporosis drugs in the osteoporosis patients can effectively improve the efficacy and reduce the relative complications.
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AIM: To investigate the effects of PAR-2 agonist peptide on the proliferation and cytosolic calcium concentration ([Ca~(2+)]_c) in human hepatoma cells HepG2. METHODS: Human hepatoma cell line HepG2 was cultured. The cells were treated with PAR-2 agonist peptide SLIGKV-NH_2 and the reverse PAR-2 agonist peptide VKGILS-NH_2, respectively. The [Ca~(2+)]_c of hepatoma cells were measured by microfluorimetric techniques based on calcium indicator fura-2/AM. The influences on proliferation of hepatoma cells were determined by MTT method. The changes of cell cycle were evaluated by flow cytometry, and the changes of cyclin D1 mRNA expression were detected by RT-PCR. RESULTS: After treated with 50 μmol/L SLIGKV-NH_2, a rapid rise of [Ca~(2+)]_c in HepG2 cells was induced (P<0.01), percent S phase, G_2/M phase and proliferation index (PI) of HepG2 cells were elevated (P<0.01), and cyclin D1 mRNA expression was significantly upregulated (P<0.01). The proliferation rates of HepG2 cells treated with 1-50 μmol/L SLIGKV-NH_2 were significantly increased, and the effect was in a dose-dependent manner (P<0.01 or P<0.05). No statistical significance of the difference between VKGILS-NH_2 and control group was observed (P>0.05). CONCLUSION: PAR-2 agonist peptide induces the rise of [Ca~(2+)]_c in HepG2 cells, upregulates the expression of cyclin D1 mRNA, accelerates the progress of cell cycle, promotes the synthesis of DNA and the proliferation of hepatoma cells via activating PAR-2 in vitro.
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Objective To study the expression of tyrosine protein kinase in hepatocellular carcinoma(HCC)and observe the correlation between tyrosine protein kinase and clinical features.Methods A total of 30 cases with HCC were enrolled in this study.ERK,P38,C-jun,JAK2,STAT3 and STAT5 were detected by immunohistochemical method using tissue chip technology.Results The expression of ERK(0.220±0.033),P38(0.174±0.024),C-jun(0.183±0.064),JAK2(0.192±0.044),STAT3(0.197±0.078)and sTAT5(0.181±0.066)in HCC was significantly higher than that(0.065±0.028,0.058±0.028,0.042±0.016,0.070±0.030,0.052±0.024,0.052±0.023)in cirrhosis 1iver tissues(P＜0.01).There was significantly positive correlation of the expression between ERK,C-jun,JAK2,STAT3 and STAT5(P＜0.01 or P＜0.05).But the expression of P38 was negatively correlated with ERK in the HCC tissues(r=-0.404,P＜0.05).JAK2 had significant correlation with tumor differentiation.The expression of J AKz in stage Ⅲ was significantly higher than that in stage Ⅰ and Ⅱ cancer tissues.Conclusion There is important significance of the excessive activation of MAPK and JAK-STAT signaI transduction in hepatocellular carcinoma process.The unbalance of signal transduction might be one of the pathogenesis of tumor progress.
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0.05). Conclusion Matrine could significantly down-regulate the mRNA expression of stat3 and stat5 and protein expression of STAT3 and STAT5 in SMMC-7721 cells. So matrine could inhibit the signaling transduction pathway of JAK-STAT and inhibit the proliferation of SMMC-7721 cells.