Objective To establish the quality standard for Zaizao Nongsuo Wan. Methods Artificial Calculus Bovis, Radix Paeoniae Rubra, Rhizoma Coptidis and Herba Ephedrae were identified by TLC. The content of Muscone was determined by GC. Results Under the selected conditions, the clear and distinguished spots were obtained. For content determination, the linear range of Muscone was 0.101 2 ? g～ 1.012 ? g, r=0.999 9, the average recovery was 99.11 % ( n=5) and RSD was 3.59 % . Conclusion The methods are proved to be simple, specialization with a good reproducibility, and can be used to control the quality of Zaizao Nongsuo Wan.

OBJECTIVETo find the characteristic parameters from electrocardiogram (ECG) which is most related to pathological progress, surface ECG was performed in mice at 2, 5, 9 and 13 week post surgery.

METHODSElectrocardiogram recordings: The mice were anaesthetized with tribromoethanol (250 mg/kg, i.p.). Lead II surface ECG were acquired by using the Biopac System MP150 at a rate of 5 kHz.

RESULTS(1) No arrhythmia was observed in Sham and 2-week Band mice. Spontaneous arrhythmias were observed in ECG recordings, with an incidence of 15%, 28% and 63% in 5-, 9- and 13-week Band mice, respectively. Tachyarrhythmias, such as frequent premature ventricular extrasystole, bigeminy, trigeminy, and paroxysmal tachycardia were found. (2) Analysis of ECG recordings revealed a significant prolongation of QT and QTc intervals. Compared with age-matched Sham mice, the increment in QT and QTc intervals was 20.4%, 32.7%, 49.7%, 61.0% and 27.1%, 32.1%, 43.9%, 59.1% at 2, 5, 9 and 13 week, respectively (P < 0.01). (3) The characteristic change in electrocardiogram was on the J wave. The amplitude of J wave was upward in Sham mice, and it was significantly flattened or inverted in Band mice. (4) Except that RR interval at 2 week showed a slight decrease, there was no significant change in PR interval and RR interval in Band mice.

CONCLUSIONIn the present study, we found that the increase incidence of spontaneous arrhythmias, prologation of QT interval and changes in the amplitude of the J wave in the surface ECG during progression of mouse cardiac hypertrophy and failure, and the progressive pattern indicated that a gradual aggravation of the ventricular repolarization delay in this mouse model.

Animals ; Cardiomegaly ; physiopathology ; Disease Models, Animal ; Electrocardiography ; Male ; Mice

OBJECTIVETo explore the alterations of after depolarization and triggered activity in myocardiac muscle during progression of mouse aortic stenosis.

METHODSAfter the establishment of mouse aortic stenosis model, the heart was collected and intracellular potential was recorded using standard glass microelectrode. After the recording, the action potential (AP), early after depolarization (EAD), delayed after depolarization (DAD) in papillary muscles were analysed.

RESULTS(1) Compared with that of time-matched control mice, there were no changes of the action potoutial duration at 90% repolarization(APD90) in model mice at 2 and 5 weeks, whereas the APD90 was progressively prolonged at 9 and 13 weeks. (2) During the 30 minites of the experiment, spontaneous EADs and DADs were frequently present in medel mice at 9 and 13 weeks, were not in control and model mice at 2 and 5 weeks. (3) Compared with those of control group, EADs and DADs triggered by hypokalaemia or isoproterenol were increased significantly. The incidence rate of triggering EADs and DADs was progressively increased over time.

CONCLUSIONEAD, DAD and trigger activities increase gradually during the process of aortic stenosis, and it suggests that the myocardium electrical-instability is markedly increased in the insult hearts.

Action Potentials ; physiology ; Animals ; Aortic Valve Stenosis ; physiopathology ; Disease Models, Animal ; Male ; Mice ; Myocytes, Cardiac ; physiology

Icarrin is the main monomer extracted and purified from Epimedium koreanum Nakai. Using dependentcell lines and LDH release assay,the synergistic effects of icarrin on inducing IL-2, 3, 6 and enhancing NK,LAK cell activity were studied. Results showed that icarrin can induce tonsil mononuclear cells to produce IL-2, IL-3,and IL-6 in collaboration with PHA in a dose-dependent manner. IL-2 and IL-6 peaked at 48h after reaction,and IL-3 peaked at 72h. Icarrin enhanced NK and LAK cell activity dose-dependentlylthe LAK activitypeaked on the 5th day. These results suggested that icarrin may be an effective BRM agent

Acupuncture Points ; Administration, Cutaneous ; Adolescent ; Adult ; Bloodletting ; Drugs, Chinese Herbal ; administration & dosage ; Ear ; blood supply ; Female ; Hordeolum ; drug therapy ; therapy ; Humans ; Male ; Young Adult

Objective To investigate the effect of Bunao capsule on learning,memory and antioxidative abilities of rats with Alzhheimer’s disease(AD)induced by D-galactose combined with amyloid β-protein(Aβ25-35 ),and provide experimental basis for the prevention and treatemtn of AD. Methods A total of 90 SD male rats were randomly divided into model control group,piracetam group,sham operated group,Bunao capsule(0.79,1.58,3.15 g·kg-1 )groups(n= 15 each).The rat models were established by intraperitoneal injection of D-galactose and injection of Aβ25-35 into the bilateral lateral cerebral ventricle.Then rats were given corresponding drugs by gavage in different groups for 8 weeks.The learning and memory abilities were meseured by Morris water maze test.The morphology of brain cells was observed by HE staining.The activities of glutathione peroxidase(GSH-Px)and superoxide dismutase( SOD),and the malondialdehyde( MDA)contents in the brain tissues were measured by spectrophotometry. Results The target quadrant residence time was(20.39±7.75)s and(20.82±5.09)s in Bunao capsule (1.58,3.15 g·kg-1 )groups,which were significantly increased as compared with that in model control group[(12. 35 ± 6.95)s](P<0.01).Brain nerve cell morphology in Bunao capsule(1.58,3.15 g·kg-1 )groups was obviously improved as compared with that in model control group,and was close to that in sham operated group.The activities of GSH-Px and SOD were significantly increased,and MDA contents decreased in Bunao capsule groups as compared with those in model control group (P<0.01). Conclusion Bunao capsule can dose-dependently improve the learning,memory and antioxidative abilities of AD rats.The mechanism may involve upregulation of antioxidative enzyme activities and removal of oxidative products.

Objective To assess and analyze the current status of clinical guidelines for hypertension in the world by using the Appraisal of Guidelines for Research and Evaluation (AGREE) instrument.Methods The clinical guidelines for hypertension were identified and approved by searching China hownet,WANFANG database,PUBMED database,MEDLINE,Embase and related institutions and authorization web site from 1995 to January 2012,and relevant Web sites of agencies and organizations that produce and/or endorse guidelines.Names of the guidelines,published years and organizations,methodology of development and reference number were descriptively analyzed.AGREE instrument was used to evaluate the qualities of latest edited clinical guidelines for hypertension in countries all over the world.Results Nine guidelines were enrolled.The results showed that the hypertension guidelines scored the highest average of 88.4％ for clarity of presentation and reliability field; for applicability fields,scored an average of 86.1％; the scope and goal field scored an average of 83.8％; participants field scored an average of 71.7％; editorial independence field scored an average of 64.1％ ; rigor of development field scored the lowest average of 62.9％.The overall assessment showed that NICE 2011,Canada 2012,ESC 2009,Australia 2010 editions were the positively recommended guidelines,JNC7,Japan 2009,China Taiwan 2010,China 2010,South Africa 2011 editions were the recommended guidelines (still need to supplement and improve).No recommend or uncertain guide was found.Conclusions The quality of the hypertension guidelines is higher in general,but some common deficiency in the rigor of development and editorial independence in Asian and African guidelines formulated by the states still exists.There still exist certain gaps in evidence-based medical requirement.And the contents and quality are needed for further regulating and enhancing.A set of scientific systemic hypertension clinical guidelines evaluation system should be established.

Objective To investigate the effect of Toll-like receptor 2(TLR2)on the proliferation of human keratinocytes.Methods Keratinocytes were isolated from the foreskin of children,and subjected to primary culture.Atier 3-5 passages.the kemtinocytes were incubated with various concentrations of peptidoglycan(PGN).a TLR2 agonist.Cell proliferation was detected by MTT colorimetric assay and the suitable concentrations of PGN were determined.The mRNA and protein expressions of Ki67.TLR2.NF-kB p65 and TGF-α were detected by real-time quantitative PCR and Western blot.respectively,in keratinocytes treated witll PGN of 0,1.25,2.5 and 5 μg/mL.Antibody blocking test was utilized to evaluate the effect of blocking TLR2 with specific anti-TLR2 neutralizing monoclonal antibody before incubation with PGN on the expressions of Ki67,TLR2,NF-KB p65 and TGF-α by keratinocytes.Results The proliferation of kemtinocytes was significantly promoted by the incubation with PGN of 1.25,2.5 and 5μg/mL for 24 hours (all P＜0.05),which also increased the expression of Ki67 protein,TLR2 mRNA and protein,and NF-KB p65 protein.Further more,the mRNA expression of Ki67 in keratinocytes was elevated bv PGN of 1.25 and 2.5μg/mL,the mRNA expression of NF-KB p65 elevated by PGN of 1.25μg/mL,and the expressions of TGF-αprotein and mRNA elevated by PGN of 1.25 and 5μg/mL (P＜0.05).The mRNA and protein expressions of Ki67,TLR2,NF-kB p65 and TGF-αwere all inhibited by the blocking of TLR2 before incubation with PGN (a11 P＜0.05).Conclusion Activation of TLR2 bv PGN could induce the over-proliferation of human keratinocytes,likely through promoting NF-rB activation and TGF-α expression.

Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

Objective To explore the biological characteristics and karyotype of bone marrow-derived mesenchymal stem cells(MSCs)in patients with systemic lupus erythematosus(SLE)and hematopoietic sup- port of MSCs.Methods MSCs were isolated from bone marrow of 11 SLE patients and 6 healthy controls by density centrifugation and adhesive culture in vitro.The surface markers were detected by flow cytometry (FCM).The morphological changes of MSCs were observed in primary and passage cultures.The growth curves were assayed.The karyotype of MSCs was detected by blocking cellular mitosis with colchicines.The MSCs from SLE patients and healthy controls were infused to ICR mice after high-dose chemotherapy.The changes of peripheral blood counts of the mice were recorded.Results Approximately(6～9)?10~9 MSCs from SLE were obtained after 5 passages and their growth was slower than normal controls(P＜0.01).Both groups were positive for CD29,CD44 and CD105,and negative for CD14,CD34,CD45 and HLA-DR.MSCs from SLE had a normal karyotype.MSCs infusions of the two groups were accompanied by no adverse event and the recovery of white blood cell,hemoglobin and platelet count was quicker when compared with the controls(P＜0.05).Conclusion MSCs from SLE have demonstrated abnormalities in expansion in vitro.MSCs from SLE have a normal karyotype.Ex vivo MSCs infusion from SLE patients can support hematopoiesis as normal MSCs.