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OBJECTIVE To evaluate the quality of total flavonoids of Lamiophlomis rotata from different producing areas. METHODS Total flavonoids of S1-S15 batches of L. rotata from different producing areas were extracted by percolation and purified by polyamide column. The content of total flavonoids was determined by UV spectrophotometry， and its purity was calculated. HPLC fingerprint chromatograms and control fingerprint of total flavonoids from 15 batches of L. rotata were established with Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 version）. Their similarities were analyzed. Cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis were used to evaluate the quality of total flavonoids from 15 batches of L. rotata， and the main components which affected the quality of total flavonoids were analyzed. RESULTS The purities of total flavonoids from 15 batches of L. rotata were 55.82%-94.12%， with an average value of 77.72%； a total of five common peaks were identified in the fingerprint， and No. 3 peak was luteolin； the similarities between the fingerprint of each batch of samples and the control fingerprint were 0.925-1.000. By cluster analysis， S1 and S3-S9 were clustered into the first class， which were samples from Qinghai Province and Tibet Autonomous Region. S14 and S15 were clustered into the second class， which were samples from Yunnan Province. S10-S13 were clustered into the third class， which were all samples from Sichuan Province. S2 was clustered into the fourth class. The principal component analysis showed that the qualities of samples from the first and fourth classes were better； peaks 2， 3 and 5 were identified as the main components that caused the differences among different batches of samples by orthogonal partial least squares-discriminant analysis. CONCLUSIONS The qualities of total flavonoids of L. rotata from Qinghai Province and Tibet Autonomous Region are better.
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OBJECTIVE：To study the effects and active ingr edients of Tibetan medicine Lamiophlomis rotata against rheumatoid arthritis （RA）. METHODS ：Fifty-six SD rats were randomly divided into normal control group （0.5％ sodium carboxymethylcellulose），model group （0.5％ sodium carboxymethylcellulose ），methotrexate group （positive control group ，3 mg/kg），L. rotata water extract low-dose ，medium-dose and high-dose groups （0.5，1，2 g/kg，by crude drug ），L. rotata total flavonoid group （200 mg/kg，by flavonoid extat ），with 8 rats in each group. Except for normal control group ，other groups were given Freund ’s complete adjuvant （FCA）on the rat ’s right hind footpad to induce adjuvant-induced arthritis model. The next day after injection of FCA ，rats in all groups were given relevant medicine intragastrically ，once a day （once every 3 days for methotrexate group ）for 30 days. At 15th and 30th day of administration ，the degree of paw swelling of left hind foot was measured，and the arthritis index ，spleen index were calculated. At the end of 30th day of administration ，the levels of TNF-α，IL-1β， IL-6 and IL- 10 in rat serum were determined by ELISA assay ，the thymus index and spleen index were calculated ，the pathological changes of the ankle joints were observed by HE staining. RESULTS ：Compared with normal control group ，degree of paw swelling，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β， msxmX0146） IL-6 in serum at 30th day of administration were significantly com increased in model group （P＜0.01）；while the thymus indexyunbinji- and IL- 10 level at 30th day of administration were ang@swu.edu.cn significantly decreased（P＜0.01）；synovial cell proliferation and infiltration of articular cavity were observed in ankle joint. Compared with model group ，degree of paw swelling ，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β，IL-6 in serum at 30th day of administration were significantly decreased in each medicine group （P＜0.01）；while the thymus index and IL- 10 level at 30th day of administration were significantly increased （P＜0.01）；the pathological changes of arthritis were significantly improved. Compared with L. rotata water extract high-dose group ，there were no significant differences in degree of paw swelling at 15th day of administration as well as the arthritis index ，spleen index ，levels of inflammatory cytokines and pathological changes of ankle joint in L. rotata total flavonoid group （P＞0.05）. CONCLUSIONS ：Tibetan medicine L. rotata shows well anti-RA activity ，and total flavonoids may be the active ingredients of its efficacy.
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BACKGROUND:Pretreatment is a dominant way to increase the histocompatibility of biomaterials,but many pretreatments are not satisfactory due to some reactions,such as calcification,cytotoxic reaction and reducing the tension resistance.OBJECTIVE:To investigate the effect of immunogenicity,scaffold tension and cell growth factor on the non-pretreated scaffold biomaterials,and to prepare the homograft decellularized scaffolds.DESIGN:Contrast observation for the histomorphology.SETTING:The experiments were carried out in the Department of General Surgery,Affiliated Hospital of Nantong University from June 2006 to March 2007.MATERIALS:Wistar rats of SPF grade were adopted in this study.Sodium dodecyl sulfate from BioteehGrade Company(U.S.A.);basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) from Peprotech Company(UK);dynamometer from Suzhou Electrical Element First Factory (China).METHODS:The vena cava inferior from rats were selected as experimental materials,we decellularized the epithelial cells from vein according to the improved Booth's methods with low osmotic Tris buffer containing sodium dodecyl sulplate,and then reserved the extracellular matrix.After fixing,the decellularized scaffold was stained by hematoxylin and eosin,the collagen fibers were observed and photographed by light microscope and scanning electron microscope.Changes of the intensity of the scaffolds were measured before and after decellularization.The deceilularized scaffolds were transplanted subcutaneously at homogeneity rats to observe whether the scaffolds had induced immunological rejection.In combination with bFGF and/or VEGF,the scaffolds were transplanted subcutaneously at homogeneity rats and took out two weeks later,in order to exam the new vessels in the scaffolds by immunohistochemistry.Dynamometer was applied to compare the change of the intensity of the scaffolds before and after transplantation.MAIN OUTCOME MEASURES:Change of the intensity of the scaffolds after decellularization and transplantation.Formation of new vessels in scaffolds.Local rejection of the scaffold subcutaneously transplanted into rats.RESULTS:The endothelial cells of the vein were completely removed when incubated with Tris buffer containing 0.03% sodium dodecyl sulfate for 48 hours.Results of hematoxylin and eosin staining,light microscope and scanning electron microscope showed that,the main components of extracellular matrix (collagen fiber) were reserved.There was no significant difference in the morphological structure and the intensity of collagenous scaffolds before and after decellalarization (P＞0.05).There was no apparent reject reaction at the embedded site,local incisions healed well.New vessels could be observed in the scaffolds two weeks later.There was no significant difference in the intensity of collagenons scaffolds before and after inoculation (P＞0.05).CONCLUSION:The low osmotic Tris buffer containing 0.03% sodium dodecyl sulfate is suitable for decellularizing the vein.The homograft collagenous scaffolds exhibit no rejection after transplantation,and the intensity of the scaffolds does not descend.VEGF and bFGF can improve the growth of new vessels in the decellularized scaffolds,and have a synergistic effect.
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Objective To evaluate the alterations of thromboxane (TXB_2)、 prostacyclin (PGI_2)in murine acute pancreatitis (AP) and the therapeutic effect of calcium channel bocker-verapamil. MethodsA rat AP model was established to observe the alterations of plasma levels of TXB_2, 6-keto-PGF_ 1? and TXB_2/6-keto-PGF_ 1? ratio and the effect of verapamil. Pancreatic histology was examined by light and electron microscopy. ResultsAt 16 and 24 hours after induction of AP, the plasma levels of TXB_2[(1112?235)pg/ml、(1265?162)pg/ml] and TXB_2/6-keto-PGF_ 1? ratio(9.9?0.9,10.2?1.3)increased significantly. With varapamil therapy, the plasma levels of TXB_2[(671?102)pg/ml、(697?93)pg/ml] and TXB_2/6-keto-PGF_ 1? ratio (6.9?2.2)、(6.4?0.7) were dramatically lower than those in control group (P
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Objective To investigate the effect of lymphatic flow impedement on intestinal bacterial translocation in rats with obstructive jaundice. Methods Murine obstructive jaundice model was established by ligating lower choledochus. Sixty rats were divided into three groups: sham-operative group (group A), obstructive jaundice group (group B), and obstructive jaundice with thoracic duct ligation group (group C). At post-op day 15 vena cava inferior blood? portal vein blood were drawn, and endotoxin, TNF-?, IL-6 were determined. Baterial culture of mesenteric lymph nodes and lung were measured with aseptic manipulation. The extreme intestinal lumen mucosa and lung were studied pathologically. Results Blood concentration of endotoxin increased in group B and C ( P