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.

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

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

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

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.

Objective To explore the clinical features and treatment of eosinophilic lymphoid granuloma(ELG) in children,in order to increase understanding of the disease.Methods Clinical features,laboratory examination,pathological examination,treatment and the-(rapeutic) effect of 5 patients with ELG were retrospectively studied,and the related literature were reviewed.Results Five cases were all boys.They all presented many subcutaneous swellings and lymphadenopathy.A case among them was associated with nephrotic syndrome,and 2 cases had the complication of eczema repeatedly,and 1 case ever had 2 times episodes of asthmatiform bronchitis.All of them had no hepatomegaly and splenomegaly,physically was well developed.Only chemotherapy was supplied to all the patients.During the course of therapy,4 cases among them recurred.After that,they were successfully treated with irradiation of lesion or cyclosporine(CsA).Conclusions ELG in children is as same as adult in clinicopathological features.It tends recur only with chemotherapy.We can get better curative effect with the synthetic treatment of chemotherapy,radiotherapy,operation or oral CsA and so on.

0.05). But the LDH 5 and LDH 5/LDH 1 of difference doses of Shenmai Injection group was lower than that of the control group obviously(P

Objective To elucidate the ESI-MS performance of lipophilic tanshinones and to establish a fingerprint of lipophilic tanshinones of Radix Salviae Miltiorrhiza.Methods The lipophilic tanshinones were extracted by ultrasonic wave with 95% ethanol from Radix Salviae Miltiorrhiza and the extracts were analyzed directly in positive ion mode by electrospray ion trap mass spectrometry(ESI-MS).Results The lipophilic tanshinones were easily to form molecular ions ~+ and dimeric sodium adduct ions ~+ in positive ion mode,molecular ions of lipophilic tanshinones were fragmentated through lossing H_2O,CO and A-ring cleavages in ESI-MS~2.The ESI-MS fingerprints of lipophilic tanshinone extracts with 14 selected characteristic peaks analyzed with SPSS software were characteristic and stable.Conclusion Lipophilic tanshinones have similar ESI-MS performance;the ESI-MS fingerprint is a useful tool for a rapid and special identification of lipophilic tanshinones in selected traditional Chinese medicine.