Objective To observe the effect of ?-asarone combined with eugenol against PC12 cell injury induced by amyloid beta protein (A?25-35) and to explore the advantage of compatibility of monomers in herbs.Methods PC12 cells injury were induced by A?25-35.The morphological features and survival rate of PC12 cells as well as the changes of intracellular Ca2+concentration and NO concentration were observed after administration.Results The optimum combination was 10?mol/L ?-asarone+3 ?mol/L eugenol,which can decrease the concentration of intracellular Ca2+,inhibit the production of NO and increase the survival rate of PC12 cells.Conclusion The combination of effective components in Acorus tatarinowii has a better protective effect on A?25-35-induced PC12 cell injury than Acorus tatarinowii.

AIM:To explore the effect of β-asarone on vascular endotheliam and adhesion molecule expression of endothelium induced by β-amyloid peptide from Alzheimer's disease and to estimate the injury repair.METHODS:Cultured ECV304 cells were incubated with freshly solublizeal Aβ1-42 and the mixture of Aβ1-42 and β-asarone,the expression of three central adhesion molecules,CD106,CD62P,CE62E and Ca2+ concentration were examined and apoptosis was recorded by Flow eytometry.Test viability of cells by MTT methods.RESULTS:The results showed that in model group and treated group,ligation of endothelial CD106,CD62P,CE62E,markers for endothelial cell activation and Ca2+ concentration,leads to a lot of release.The livability decreased and the apoptosis increased.Further more,simultaneous treatment of ECV304 cells with β-asarone resulted in the decrease significandy in these three adhesion molecules described above and Ca2+ concentration as well as the livability upper and apoptosis lower.CONCLUSION:CD106,CD62P,CE62E,important inflammational factor of Aβ-induced endothelial injury,may be promotion of the inflammatory scade in vascular endothelial.β-asarone may protect ECV304 cell apoptosis by regulate Ca2+ and expression of cell surface markers.

AIM:To explore the effect of ?-asarone on vascular endothelium and adhesion molecule expression of endothelium induced by ?-amyloid peptide from Alzheimer's disease and to estimate the injury repair.METHODS:Cultured ECV304 cells were incubated with freshly solublized A?_ 1-42 and the mixture of A?_ 1-42 and ?-asarone,the expression of three central adhesion molecules,CD106,CD62P,CE62E and Ca 2+ concentration were examined and apoptosis was recorded by Flow cytometry.Test viability of cells by MTT methods.RESULTS:The results showed that in model group and treated group,ligation of endothelial CD106,CD62P,CE62E,markers for endothelial cell activation and Ca 2+ concentration,leads to a lot of release.The livability decreased and the apoptosis increased.Further more,simultaneous treatment of ECV304 cells with ?-asarone resulted in the decrease significantly in these three adhesion molecules described above and Ca 2+ concentration as well as the livability upper and apoptosis lower.CONCLUSION:CD106,CD62P,CE62E,important inflammational factor of A?-induced endothelial injury,may be promotion of the inflammatory scade in vascular endothelial.?-asarone may protect ECV304 cell apoptosis by regulate Ca 2+ and expression of cell surface markers.

Objective To observe the effect of β-asarone on the proliferation, cycle, apoptosis and migration of tumor cells A549, PC3, and PC9-R, thus to provide experimental basis for the application of β-asarone to the treatment of cancer. Methods After A549, PC3, and PC9-R were cultured with different concentrations ofβ-asarone for 24 h, 48 h, and 72 h respectively, CCK-8 was used to detect the optical density (D) of cell proliferation, and then the cell proliferation rate was calculated. The flow cytometry was used to measure the cell DNA cycle and cell apoptosis, and Transwell Chambers were used to detect the cell migration. Results After treatment with different concentrations of β-asarone for 24 h, 48 h, and 72 h respectively, the growth of A549, PC3, and PC9-R showed declining trend in concentration- and time-dependent manner. The proportion of A549, PC3, and PC9-R at G0/G1 phase was increased, the proportion of the three kinds of cells at S phase and the proliferation indexes were declined, the apoptotic rate of A549, PC3, and PC9-R was increased, and the migration of A549, PC3, and PC9-R was reduced (P<0.05 or P<0.01 compared with those of the normal control group). Conclusion β-asarone has certain effects on suppressing proliferation, blocking G0/G1 phase developing into S phrase, inhibiting DNA synthesis, promoting the apoptosis, and inhibiting the migration of A549, PC3 and PC9-R.

Objective To investigate the effects of ?-asarone,the major constituent of Acorus tarinowii schott,on the expression of endothelial cell adhesion molecules induced by hypoxia and reoxygenation.Methods The expression rates of endothelial cell surface adhesion molecules VCAM-1 and CD62E were detected by flow cytometry(FCM) and specific labeled monoclonal antibody.Results Compared to the normal group,the expression rates of VCAM-1 and CD62E were increased significantly in the model group(hypoxia-reoxygenation group)(P

Object To distinguish the genuine Dioscorea polystachya Turcz. from other species of Dioscorea L. such as D. persimilis Prain et Burkill, D. japonica Thunb., and D. alata L., conventionally used in some localities by sequencing their nuclear ribosomal RNA subunit (18S rRNA), to provide mole cular evidences for the quality control of drugs of Dioscorea L. Methods By direct PCR sequencing to detect the homology of 18S rRNA sequence of different species of Dioscorea L.. Results The 18S rRNA gene sequence of genuine D. polystachya, D. persimilis and D. japonica showed identical length of 1810 bp, whereas that of D. alata was 1807 bp. Multiple sequence alignment of 18S rRNA gene showed that the homology was identical between D.persimilis and the genuine D. polystachya; but a difference of 99.89% with D. japonica and 97.51% with D. alata. Conclusion DNA sequencing can provide an accurate and reliable mean for the identification of the origin and genuineness of Dioscorea L..

A strain of Prototheca species isolated from a case of meningitis was identified by routine morphologic and biochemical methods as well as amplification of the related genes, in which the 28S large-subunit (LSU) region of ribosomal RNA (rRNA) gene and intergenic space (ITS) were amplified with universal fungal primers. The small-subunit (SSU) rRNA gene was amplified with eukaryote-specific primers and Prototheca genus-specific primers. Then, compared the sequences with the ones posted on BLAST (www. ncbi. nlm. nih. gov/BLAST). The organism choice giving the closest match, up to 99%, was considered the most likely correct identification. It was found that this strain of fungus grew well at 25 ℃ or 37 ℃. Smooth,moist colonies with white color were observed on Sabouraud Dextrose Agar (SDA) and Potato Dextrose Agar (PDA). Microscopically, globular or ovoid cells, a number of round, ovoid shaped endospores could be observed. No hypha, ascus or blastic conidia was found upon cultivation on SDA. Based on the morphological characteristics, this isolate could be identified as Prototheca species. The identity with Prototheca wickerhamii was 2.9 % as demonstrated by the API 20C AUX system. Sequence analysis showed that the ITS gene was proved to be a complex structural region which was not suitable for the identification of Prototheca species, but the LSU and SSU rDNA regions showed 94% and 99.9% sequence identities with Prototheca zopfii var. hydrocarbonea (P. zopfii var. hydrocarbonea) respectively, indicating that the SSU rRNA gene sequence might be more reliable on than the LSU rRNA gene sequence for identification of Prototheca species.

Objective To investigate the effect and nursing experience of iliacus groin approaches and internal fixation treatment for patients with pelvic fractures. Methods Twenty- six patients with unstable pelvic fracture were obtained iliac groin approaches and internal fixation treatment, the changing symptoms in patients were monitored frequently, and all patients were performed general nursing. Results All patients were followed up for 0.5 to 5.0 years, without obvious complications, the results of surgical treatment included optimal 16 cases, good 7 cases, mean 3 cases. Conclusions Iliacus groin approaches and internal fixation with reconstruction plate treatment on pelvic fractures was proved to be reset accurate, reliable and strong, with better effects. Effective nursing measures can save life of patients with pelvic fractures, speed up rehabilitation of physiological function, improve life quality of the patients.

【Objective】To observe the effect of ligustrazine(Lig) on cell apoptosis and expression of bcl-2 and p53 in rats hippocampal neurons after anoxia,and to explore its protective mechanism.【Methods】Primary culture technique of rat hippocampal neurons was used to establish the model of anoxia.At the 14th day of the culturing,the cultured cells were divided into model group,and high-,middle-and low-dosage ligustrazine (800,200 and 50 ?g/mL respectively) groups,which were exposed to anoxia environment for continuous 1.5 hours.Flow cytometer was used for the quantitative analysis of apoptosis rate of neurons,and in-situ hybridization was used to detect the bcl-2 and p53 mRNA expression of hippocampal neurons.【Results】In middle-dosage ligustrazine group after exposure to anoxia environment for 1.5 hours,the apoptosis rate and the bcl-2 and p53 mRNA expression of hippocampal neurons differed from those in other groups(P

Objective To observe the effects of Tianma Gouteng Yin (TGY) on the proliferation of myocardial fibroblasts of spontaneous hypertensive rat(SHR) stimulated by insulin. Methods The cardiac fibroblasts from one-day-old spontaneously hypertensive rat were primarily cultured and then sub-cultured to 3rd-6th generation,and were identified with light microscopy and immunohistochemical technique. The effect of serum containing TGY on the proliferation of cardiac fibroblasts of SHR treated with insulin was determined by MTT colorimetric assay and 3-H-TdR incorporation,and the cell cycle was analysised with flow cytometer. Results TGY inhibited the proliferation of cardiac fibroblasts induced by insulin. Insulin can stimulate the proliferation of fibroblasts by promoting the cell cycle transfer from G0/G1 period to S period and increasing the synthesis of cell DNA. Compared with the model group,the percentage of the cells at G0/G1 period was much higher,while the percentage of the cells at S period was much lower in TGY group. Conclusion TGY has the effect on inhibiting the proliferation of cardiac fibroblasts and inhibiting the cell cycle transfer from G0/G1 period to S period,which may be one of its therapeutic and protective mechanisms for hypertension and myocardial fibrosis in clinic.