Objective To establish the fingerprint of Yuanshi Shengmai Chenggu (YSC)Tablets by HPLC.Methods Chromatographic conditions of HPLC were as follows: Hypersil ODS2 column with temperature at 20 ℃; methanol and 1 %acetic acid glacial(gradient elution)as a mobile phase; detection wavelength at 268 nm;analytical time being 55 min and flowing rate being 1.0 mL/min.Results Twenty-five peaks were indicated on the HPLC-fingerprint of YSC Tablets. The peak area of vitexin was the biggest area in different batches of YSC Tablets, the area being 22 %～25 %. The differences of fingerprint in different batches of YSC Tablets were not obvious, which indicated the fingerprint characteristics of YSC Tablets.Conclusion The method is simple and accurate and with a good reproducibility and can be used for the quality control of YSC Tablets.

Objective To compare the differences between external standard method and relative correction factor method for determination of the flavonoids from Sorbus tianschanica Rupr.Methods Using HPLC external standard method for determination of hyperoside,rutin,isoquercitrin,quercetin-3-O-(6″-O-malonyl)-β-D-glucoside,astragalin and Kaempferol-3-O-(6″-O-malonyl)-β-D-glucopyranoside in Sorbus tianschanica Rupr.,HPLC relative correction factor method was adopted to establish relative correction factor of the other five flavonoids above with hyperoside as reference.The difference was evaluated by comparing the external standard method with the relative correction factor method.Results There was no significant difference between the T test external standard method and relative correction factor method(P>0.05).Conclusion External standard method and relative correction factor method can be used for determination of the flavonoids from Sorbus tianschanica Rupr.,but in the case of lack of reference substance or mass detection,using the relative correction factor method for determination of rutin,hyperoside isoquercitrin,quercetin-3-O-(6″-O-malonyl)-β-D-glucoside,astragalin and kaempferol-3-O-(6″-O-malonyl)-β-D-glucopyranoside in Sorbus tianschanica Rupr.It was more feasible and it can be used as a new quality evaluation method in determination of flavonoid components from Sorbus tianschanica Rupr.

Aim To explore the effect of adenylate cyclase(AC) antagonists SQ22536 and agonist forskolin on acute lung injury induced by lipopolysaccharide.Methods ICR mice were randomly divided into normal saline control group(N group), model group(group L), dexamethasone group(group D),AC antagonists s(group SQ) and AC agonist group(group F).The ALI mouse model was induced by instilling intratracheally with LPS(2 mg·kg-1), and 6 h later, the lung tissue and alveolar lavage fluid(BALF) were harvested, pathological changes in lung were observed, white blood cell and neutrophil, albumin content in BALF and myeloperoxidase(MPO) activity of lung tissue homogenate were determined, and tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β), interleukin 6(IL-6) and cAMP content in lung homogenates were detected by ELISA.Results Compared with normal saline group, a large number of neutrophils infiltrated around the pulmonary vessel and airway 6 h after LPS intratracheal instillation in model group.White blood cells and neutrophils and protein content increased in BALF;MPO activity and cAMP levels increased in lung tissues.In the lung tissue TNF-α and IL-6, IL-1β content increased, compared with model group.Forskolin could improve the pathological changes of lung tissue, reduce the total number of leukocytes, number of neutrophils and protein content in BALF, and reduce MPO activity and TNF-α content in lung tissue, at the same time it increased the cAMP content;SQ22536 had no significant effect when compared with model group.Conclusion AC agonists have protective effects on LPS-induced acute lung injury in mice, and the mechanism may be related to elevating cAMP levels, inhibiting neutrophil adhesion and chemotaxis and reducing inflammatory factor levels.

Adult ; Bone Diseases, Metabolic ; etiology ; Female ; Fractures, Spontaneous ; etiology ; Humans ; Liver Diseases, Alcoholic ; complications

Objective To investigate the effect and possible mechanism of high mobility group box (HMGB) 1 in the development and progress of rheumatoid arthritis.Methods PBMC and serum samples were obtained from 74 RA patients (38 in active stage and 36 in stable stage) and 26 healthy controls.The expression of HMGB1 mRNA and protein was detected by RT-PCR and ELISA.Flow cytometry analysis ( FCM ) was used to detect the expression of Toll-like receptor 4 on PBMC.Results ①The expression of HMGBI mRNA and protein in active RA patients was significantly higher than that in healthy controls and inactive RA patients [2.63 vs 0.71，0.93 and (10.2±1.2) vs (7.5±1.8)，(8.3±1.8) ng/ml，respectively](P＜0.01 ).② The relative expression of TLR4 protein on CD14+ monocytes and CD3+ lymphocytes in active RA patients was increased than that in inactive RA and healthy controls (P＜0.05 or P＜0.01 ).It was also higher in inactive RA than in healthy controls (P＜0.05 or P＜0.01 ).③ Level of HMGB1 protein in serum of RA patients was positively correlated with ESR，CRP，RF，the numbers of tender joints and swollen joints as well as radiographic changes.Conclusion HMGB1 can be synthesized and released by PBMC of active RA patients，and then bind to TLR4 of PBMC to promote inflammatory responses and bone erosion.

Aim Toestablishanexcellentratasthmamodel from using OVA+pertussis sensitized, OVA sensitized and per-tussissensitizedrats.Methods Thethreemethodswereusedto sensitize rats;methacholine bronchial provocation tests were per-formed to determine airway hyperresponsiveness;bronchoalveolar lavage fluid ( BALF) was prepared after the animals were chal-lenged by nebulized antigen. The differential white cell count in BALF was performed, and lung tissue was detected by morpho-logicalanalysis.Results AllofOVA+pertussissensitization,OVA sensitization and pertussis sensitization could deteriorate lung function, increase inflammatory cells and cause pathological change, and OVA + pertussis sensitized rat model had better effect than OVA sensitized and pertussis sensitized rat models. Conclusion OVA+pertussissensitizationandOVAaerosolisa successful rat asthma model.

ABSTRACT:Objective To study the effects of weight and anteroposterior diameter-to-transverse diameter ratio on establishing a model of acute myocardial infarction (AMI)in rats without artificial ventilation and changes in left ventricular function after infarction.Methods Healthy SD rats were randomly divided into group A (200-250 g),group B (250 - 300 g),group C (> 300 g),and group D (control group).The left anterior descending (LAD)coronary artery was ligated to establish a model of myocardial infarction under spontaneous breathing condition immediately after thoracic lines were measured.And changes of electrocardiography were recorded after model establishment.At 2 and 4 weeks after AMI,we observed ventricular wall thickness and ventricular wall motion and measured the changes of cardiac function.Histomorphological changes and myocardial ultrastructure of the heart were observed under thoracotomy 2 weeks after operation.The above data were analyzed by SPSS13.0 statistics software.Results ① The first AMI rat model was established successfully after 30 times of experiments, and after 100 times the model’s success rate gradually stabilized at about 83%.② Group B and group C had a higher model success rate than group A (P <0.05),but group B and group C did not differ in modeling success rate (P >0.05).③ There was no association between the rate of rat thoracic line and modeling success rate (P >0.05).
④ Two weeks after thoracotomy,ischemic myocardial color was white,and ventricular wall motion decreased.HE staining revealed that cardiomyocytes disappeared and were replaced by fibrous tissues and collagen.Remnant cardiomyocytes were arranged disorderly and myofibers were fractured,with interstitial damage and hyperplasia of fibrous tissue.Visible muscle cells were sparse and dissolved,the mitochondria had darker staining,blurred cristae, and edema under electron microscopy. ⑤ Compared with group D, 2 weeks and 4 weeks after myocardial infarction,left ventricular end-diastolic diameter (LVEDD)and left ventricular end systolic diameter (LVEDs) increased (P <0.05),but EF values and heart rate dropped (P <0.05).Conclusion By this method,a model of AMI in rats can be established successfully and the heart function is changed.Under the condition of non-artificial ventilation,the weight of rats is an important factor for establishing AMI model.However,we have not confirmed the effect of thoracic lines on establishing AMI model yet.

Aim To compare the inhibition of lipid peroxidation of ethyl acetate extract( EAE) and n-buta-nol( BE) extract from Coreopsis tinctoria Nutt. in vitro. To investigate the parameters such as body weight, bio-chemical indexes in plasma, and viscera indexes on type 2 diabetes mice by intraperitoneal injection of streptozotocin ( STZ ) . Methods The extracts were prepared by response surface methodology. The ex-tracts were suspended in distilled water and defatted with petroleum ether. The aqueous layer was succes-sively extracted with ethyl acetate and n-butanol. The inhibition of lipid peroxidation activity was determined by thiobarbituric acid method. The effects of extract BE on diabetic mice were observed at the dosage of 0. 2,0. 4,0. 8 g·kg-1 ( ig) for 4 weeks. The parame-ters were observed such as weight of body changes, or-gan coefficients of liver, pancreas and kidney, bio-chemical indexes in plasma and viscera pathological sections. Results In the linoleic acid reaction system, the SC50 value of the EAE and BE was ( 443. 96 ± 11. 24) mg·L-1, (840. 29 ± 16. 38) mg·L-1, re-spectively, and that in rat liver homogenate was (23. 59 ± 3. 67 ) mg · L-1 , ( 60. 37 ± 4. 27 ) mg · L-1 , respectively. Compared with diabetic model group, BE could significantly improve the trend of weight loss, and increase viscera indexes. The patho-logical sections showed that BE had the recovery and improvement effects on the damage of liver, pancreas and kidney. Conclusions The extracts of C. tinctoria have a certain anti-lipid peroxidation activity in vitro. And BE has a certain capacity to improve and repair damaged organs for DM mice.

Objective:To investigate the effect of HMGB1 and TLR-4 in the renal injury of SLE.Methods:The level of HMGB1 in serum from 16 patients suffering from SLE without kidney damage,18 patient with lupus nephritis (LN),and 12 healthy individuals were measured by ELISA.The fresh PBMCs were isolated and the total RNA was extracted.Then the mRNA expression of HMGB1 was amplified by RT-PCR.Flow cytometric analysis(FCM) was performed to study cell surface markers and the expression of TLR-4.Results:RT-PCR and ELISA results showed that the expressions of mRNA and protein were higher in patients with LN than in SLE without kindey damage and healthy people.The expression of TLR-4 in CD14+ monocytes of patients with LN was higher either,while there were no significant changes in CD3+ T cells among LN,SLE and healthy control.Conclusion:PBMCs in patients with LN could synthesize and secrete HMGB1 initiatively,which are correlated with serum HMGB1 level.HMGB1 might play a role in autoimmunity of lupus nephritis partly by activation of monocytes through its binding to TLR-4.

Objective:To investigate the effects of TLR/STAT pathway in the proliferation of mesangial cell induced by HMGB1.Methods:Human mesangial cells were inoculated in the dose of 1?104 ml-1.After 24 h,cells were cultured with standard medium as control group or with medium supplement with 10 ?g/L human recombinant protein HMGB1 as trial group in vitro.Then the cells were collected in 6,12 and 24 h respectively,as well as control group cells.Immunocytochemical staining was adopted to examine the expressions of PCNA proteins on mesangial cells in different groups.Immunocytochemical staining and FCM were performed to detect the changes of TLR2 protein expression.STAT1 and STAT3 mRNA were examined by RT-PCR technique.Results:Immunocytochemical staining indicated that the mesangial cells could multiply after they were induced by human recombinant protein HMGB1.Immunocytochemical staining showed that the level of TLR2 protein in trial groups were higher than those in control groups.FCM indicated that HMGB1 could significantly up-regulate the expression of TLR2 protein time-dependently.The STAT1 and STAT3 mRNA in HMGB1 groups were higher than those in control groups.The expression of TLR2 protein was positively correlated with those of STAT1 and STAT mRNA respectively.The positive rate of PCNA was remarkably correlated with the expression of STAT1 and STAT3 mRNA.Conclusion:HMGB1 could activate STAT1/STAT3 through combining with its cell-surface receptor TLR2,which may play an important role in promoting the proliferation of mesangial cells and then damaging the renal of lupus nephritis.