OBJECTIVE:To establish the quality standard of Yanyan granules of concentrated sugar-free type. METHODS:Scrophularia ningpoensis,Menispermum dauricum,Radix et Rhizoma Glycyrrhizae in the granules were identified by TLC,and the content of linarin in Yanyan granules were determined by HPLC.RESULTS:The TLC spots were clear with high resolution.The linear range of linarin was 4.64～69.6 ?g(r=0.999 9) and the average recovery rate was 98.65%(RSD= 0.79%,n=6). CONCLUSION:The established standard is suitable for the quality control of the Yanyan granules of concentrated sugar-free type.

Objective To investigate the influence of poly(ADP-ribose)polymerase inhibitor PJ34 on blood brain barrier(BBB)in rats with cere-bral ischemia-reperfusion injury. Methods Rat model of cerebral ischemia-reperfusion injury was established by the middle cerebral artery occlu-sion. A total of 135 SD rats were randomly divided into 3 groups:sham-operated group(sham group),ischemia-reperfusion group(IR group)and PJ34 group(PJ34 group). 45 animals in each group were then equally divided into subgroups and the rats were sacrificed at 6 h,24 h,48 h after re-perfusion,respectively. BBB permeability was evaluated by detection of extravasated Evans blue(EB). The expression of tumor necrosis factorα(TNF-α)and matrix metalloproteinase 9(MMP-9)activity were measured by immunohistochemistry and western blot at different time points. Re?sults Compared with sham group,the contents of EB and the expressions of TNF-αand MMP-9 in IR group were increased significantly(P<0.05). Compared with IR group,the contents of EB and the expressions of TNF-αand MMP-9 in PJ34 group were markedly decreased at the same time point(P<0.05). Conclusion The present study provided in vivo evidence that PARP inhibitor PJ34 can protect against cerebral ischemia re-perfusion injury,and the mechanism might be related to maintaining the stability of blood-brain barrier by suppressing the expression of TNF-αand MMP-9 in ischemic cortex.

Objective To investigate the effects of PJ34,a poly ADP-ribose polymerase(PARP)inhibitor,on the expression of matrix metallopro-teinases-9( MMP-9 )and Claudin-5 in ischemic cortex of rats with focal cerebral ischemia-reperfusion(I/R)injury. Methods The focal cerebral ischemia-reperfusion model of middle cerebral artery occlusion(MCAO)was established by intraluminal suture . PJ34 was injected intraperitoneal-ly. Blood-brain barrier(BBB)permeability was quantitatively determined by Evans blue assay. Infarct volume changes were observed by 2,3,5-tri-phenyltetrazolium chloridedyeing(TTC)staining. The expression of the MMP-9 and Claudin-5 in rats of cerebral cortex were measured by immuno-histochemistry assay and western blot analysis . Results Compared with sham group,the expression of MMP-9,the contents of EB and infarct vol-ume increased progressively over time after I/R,and reached maximum levels at 48 h(P<0.05);The expression of Claudin-5,the contents of EB and infarct volume reduced significantly over time after I/R,and reached the minimum levels at 48 h in model group(P<0.05). Compared with model group,the expression of MMP-9,the contents of EB and infarct volume was reduced significantly and the expression of Claudin-5,the con-tents of EB and infarct volume was increased at the same time point in PJ34 group(P<0.05). Conclusion PJ34 maintained the blood-brain barri-er permeability by inhibiting the expression of MMP-9,and increasing the expression of Claudin-5,which had neuroprotection on cerebral ischemia-reperfusion injury.

Objective To study the modulation of central amygdale (CeA)on cardiac nociception with electrical stimulation in rats.Methods The heart pain model of adult male Sprague-Dawley rats was established with intrapericardial injection of capsaicin; then the evoked electromyography (EMG ) responses in the spinotrapezius (SPT)were observed as an index.Immunohistochemical method was used to observe the expression of c-fos in CeA;The rats were divided into normal control group,intrapericardial saline injection control group,and intrapericardial capsaicin injection group. The changes of EMG responses after electrical stimulation on CeA (intensity of 30μA and 70μA)were observed separately by electrophysiology in different groups.Results① Compared with normal control group, intrapericardial capsaicin injection group had significantly increased expression of c-fos in CeA.However,c-fos expression did not differ significantly between intrapericardial saline injection group and normal control group;② After 30μA electrical stimulation on CeA,EMG response in SPT significantly decreased in intrapericardial capsaicin injection group compared with that in control group;the total discharge rate was (78.58±4.62)% of that of control group (P<0.05).After 70μA electrical stimulation on CeA, the total discharge rate was (54.89±3.98)% of that of control group (P<0.05).Conclusion CeA may be involved in cardiac nociceptive transmission in rats and has inhibitory effects on the modulation of rat cardiac nociception.

Objective To investigate the effective mechanism of Jiawei Maxing Shigan Tang(JMST, Modified Decoction of Ephedra,Apricot Kernel,Gypsum and Licorice)in treatment of acute exacerbated chronic obstructive pulmonary disease(AECOPD).Methods Wistar rats(n =40)were randomly divided into blank control group,model group,JMST group and clarithromycin group.The blank control group was given normal feeding, and model group, JMST group and clarithromycin group were given airway instillation of lipopolysaccharide(LPS)and smudging for establishing AECOPD model.The model group was given normal saline,JMST group,JMST(8.7 g/kg)and clarithromycin group,clarithromycin suspension(50 mg/kg)continuously for 30 d.The lung function was measured on the 31std.The lung tissue was collected to making pathological sections for observing goblet cells after AB -PAS staining.The expressions of mucoprotein 5AC(MUC5AC)and neutrophil elastase(NE)were detected by using immunohistochemistry technique.Results Compared with blank control group, the ratio of 0.3-second forced expiratory volume to forced vital capacity(FEV0.3/FVC)decreased significantly, and number of goblet cells and expressions of MUC 5AC and NE increased significantly in model group,JMST group and clarithromycin group.Compared with model group, FEV0.3/FVC increased significantly,and number of goblet cells and expressions of MUC 5AC and NE decreased significantly in JMST group.The comparison in FEV0.3/FVC,number of goblet cells and expressions of MUC 5AC and NE had no statistical difference between JMST group and clarithromycin group.Conclusion Jiawei Maxing Shigan Tang can significantly promote FEV0.3/FVC, inhibit proliferation of goblet cells, down-regulate expressions of MUC5AC and NE and relieve airway mucus hypersecretion in AECOPD rats, which may be its one of therapeutic mechanisms in treatment of AECOPD.

Objective To explore the effect of Qingjin Huatan Decoction on forkhead box P3 (Foxp3) and retinoid-related orphan nuclear receptor γt (RORγt)in the lung tissue of airway mucus hypersecre-tion rat model with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods Altogether 40 male Wistar rats,SPF grade,were randomly divided into normal group,model group, Qingjin Huatan Decoction group and clarithromycin group.AECOPD rat model was induced by intratra-cheal instillation of lipopolysaccharide (LPS)with passive smoking.From the first day of the experiment, rats of model group were intragastrically administered saline,and those of Qingjin Huatan Decoction group and clarithromycin group took corresponding medicines at dose of 8.4 g/kg and dose of 50 mg/kg for consecutive 30 days.On day 31,all rats were sacrificed.The thin sections of the rats’resected lungs were made,the expressions of mucin 5AC (Muc5AC),neutrophil elastase (NE),Foxp3 and RORγt in lung tissue were detected by using immunohistochemical method,and the protein expressions of Foxp3 and RORγt were measured by using Western blot.Results Compared with the normal group,Foxp3 protein expression in model group had no significant difference (P >0.05),but expression of RORγt, Muc5AC and NE increased significantly with Foxp3 /RORγt decreased significantly (P <0.01 ).Com-pared with model group,Foxp3 expression in Qingjin Huatan Decoction group had no significant differ-ence (P >0.05),but expression of RORγt,Muc5AC and NE decreased significantly with Foxp3 /RORγt increased significantly (P <0.01 or P <0.05 ).Compared with clarithromycin group,Muc5AC de-creased significantly in Qingjin Huatan Decoction group (P <0.05).Conclusion The potential mecha-nism of Qingjin Huatan Decoction against AECOPD mucus hypersecretion was to downregulate RORγt protein expression in lung tissue and to upregulate Foxp3 /RORγt.

Objective This study used high-throughput sequencing technology to detect the regulation of JianPiHuaTan Prescription(JPHT)on the ApoE-/-mouse miRNA related to vascular smooth muscle cells proliferation and migration,verify the expression of related proteins,and explore the therapeutic effect of JPHT on atherosclerosis.Methods Ten of C57BL/6J mice were used as control group,and 30 ApoE-/-mice were randomly divided into model group,western medicine group and JPHT group.Sequencing technology was performed on aortic sample to select the differentially expressed miRNA.We screened out the tagetted miRNA related to VSMCs proliferation and migration.RT-qPCR and Western blot technology were used to test the differentially expressed miRNA and tagetted protein.Results Compared with model group,a total of 9 miRNAs were changed in JPHT group,among which 7 were up-regulated and 2 were down-regulated.The expression of miRNA-34a was up-regulated in JPHT group.The miRNA-34a and α-SMA protein expression in aorta of JPHT group were significantly different with model group by RT-qPCR and Western blot experiment(P<0.01).Conclusion Jianpi Huatan Prescription may improve ApoE-/-mice atherosclerosis through inhibitting the proliferation of vascular smooth muscle cells by regulating miRNA-34a.