Objective To compare optimal protein extraction method of velvet antler ( VA ) and explore the repairmen activity of velvet antler proteins ( VAPs ) on 1-methyl-4-phenylpyridinium ( MPP +) damaged nerve cell.Methods VAPs was obtained from fresh velvet antler by homogenization and micronization method respectively.The protein content of VAPs was measured by Bradford and the molecular weight was identified by SDS-PAGE.Scanning electron microscope (SEM) was perfomed to observe the ultrastructure of VAPs.Neuroblastoma cell line (SH-SY5Y) damage was induced by MPP +, and activity of each compound was compared by the proliferation of damaged cell detected by MTT method.Results The yield of micronization method of VAPs was higher than homogenization method and with better characters both in SDS-PAGE and SEM.Activity detection indicated that VAPs at the concentration of 125 μg/mL by homogenate method could significantly increase the proliferation rate of damaged SH-SY5Y cells.By contrast, VAPs by micronization method had more effective effect at 62.5μg/mL, and proliferation rate could reach 25.45% .Conclusion Micronization method does not only acquire more protein but also has cell proliferation activity, is a relatively ideal way of protein extraction of velvet antler.

A target molecule affinity-ultrafiltration liquid chromatography-electrospray ionization tandem mass spectrometric (LC-ESI-MSn) method was established for rapid screening acetylcholinesterase (AChE) inhibitors from total alkaloids of fibraurea recisa Pierre.A total of 12 potential inhibitors were screened from Fibraurea recisa Pierre.and 6 compounds were identified including palmatine,berberine,jatrorrhizine,palmatrubine,7,8-dihydro-8-hydroxyberberine and groenlandicine.The AChE inhibitory activity of these 6 compounds was validated in vitro.Palmatine showed the strongest inhibitory activity for AChE,which was stronger than that of donepezil hydrochloride,demonstrating the potential of palmatine as anti-Alzheimer's drug.This method is simple,rapid,and accurate for directly screening active ingredients which can inhibit AChE from complex extract of traditional Chinese medicines.

AIM: To investigate the relation between myocardial remodeling and the genesis of serum anti-?_3-adrenoceptor autoantibody,an animal model of heart failure(HF) was established and the biological effects of the autoantibody were observed.METHODS:(1) Healthy male Wistar rats were subjected to HF by constricting the abdominal aorta.(2) The anti-?_3-adrenoceptor autoantibody in the sera of HF rats was detected by ELISA with the synthetic peptide of the second extracellular loop of the ?_3-adrenoceptor used as the antigen.(3) IgG in the positive sera from HF rats was prepared using a MabTrap Kit(Amersham).(4) The effects of the autoantibody on the contractile response of adult(isolated) cardiomyocytes and on the beating rate of cultured neonatal rat cardiomyocytes were observed.RESULTS:(1) The positive rate of anti-?_3-adrenoceptor autoantibody of rats increased from 21.05% of pretreatment to 78.95% after heart failure(P

Objective:To determine the changes of the main components after compatibility of compound Mailuqishen(MLQS), and to explore its anti-tumor effect in the H22 tumor-bearing mice.Methods:The contents of ginsenoside and amino acid in MLQS were detected by HPLC, and the content of polysaccharide was detected by phenol-sulfuric acid.A total of 144 female Kunming mice were randomly divided into normal control group, model group, positive control group, low dose of MLQS group, medium dose of MLQS group, high dose of MLQS group, ginseng geng group, ginseng group and antler plate group (n=16).Except normal control group, the mice in the other eight groups were used to establish the H22 tumor-bearing mouse models, then the mice were treated with drugs at next day.The tumor weights, inhibitory rates of tumor, spleen and thymus indexes of the H22 tumor-bearing mice were detected 10 d after administration.The morphological changes of tumor and spleen tissue were examined by HE staining, and the apoptotic rates of H22 tumor cells were tested by flow cytometry.Results:As calculation with the ginseng and antler plate single herb, the contents of ginsenosides, polysaccharides and amino acids in MLQS were significantly higher than those of single herbs (P<0.05).Compared with model group, the inhibitory rates of tumor in various administration groups were significantly increased(P<0.01);the spleen indexes and thymus indexes of the mice in different doses of MLQS groups were significantly increased(P<0.01);the apoptotic rates of tumor cells were markedly increased(P<0.05).Compared with model group, the tumor tissue of the mice in various administration groups was destroyed, the cells were sparse and irregular, and the tumor presented necrotic lesions;the morphology of spleen tissue was normal with discernible fringe, and the lymphocytes arranged densely.Conclusion:The contents of ginsenosides, polysaccharides, and amino acids in compound MLQS are significantly increased compared with those of single herbs, and its anti-tumor effect is stronger than the single herbs.

Objective:To investigate whether ventricular non-excitatory electrical stimulation(NES)preconditioning can protect myocardial ischemia-reperfusion(IR)injury in aged mice by regulating cardiac calcitonin gene-related peptide(CGRP).Methods:Male C57BL/6J mice were randomly divided into 3 groups, (1)the Sham group, receiving sham operation group(n=6); (2)the IR group, receiving ligation of coronary artery to induce myocardial ischemia for 45 min and reperfusion for 120 min(n=13); (3)the NES group, IR model receiving NES from 15 min before IR to the end of reperfusion(n=13). Infarct size was detected by staining with 2, 3, 5-triphenyltetrazolium chloride.The level of serum cTnI and expression of myocardial CGRP were measured by enzyme linked immunosorbent assay and quantitative real time polymerase chain reaction.Results:Compared with the IR group, the infarct sizes were significantly lower in NES group[(38.17±4.36)% vs.(45.33±5.68)%, P<0.05]. Besides, the IR and NES groups were associated with significantly increased levels of serum cTnI[(10.89±2.14)μg/L, (7.03±1.79)μg/L vs.(3.92±0.47)μg/L, P<0.001], myocardial CGRP protein[(26.33±4.55)μg/kg, (19.67±5.79)μg/kg vs.(17.00±2.90)μg/kg, P<0.01], CGRP mRNA[(1.40±0.20), (2.20±0.75) vs.(1.05±0.10), P<0.01]compared with the Sham group.Furthermore, the NES group was associated with markedly decreased levels of serum cTnI and myocardial CGRP protein, and increased level of CGRP mRNA compared with the IR group(all P<0.05). Conclusion:NES may protect myocardium IR injury by regulating endogenous CGRP expression in aged mice.

The dried rattan stem of the Fibraurea Recisa Pierre plant contains the active ingredient known as fibrauretine (FN). Although it greatly affects Alzheimer's disease (AD), the mechanism of their effects still remains unclear. Proteomics and transcriptomics analysis methods were used in this study to determine the mechanism of FN in the treatment of AD. AD model is used through bilateral hippocampal injection of Aβ1-40. After successful modeling, FN was given for 30 days. The results showed that FN could improve the cognitive dysfunction of AD model rats, reduce the expression of Aβ and P-Tau, increase the content of acetylcholine and reduce the activity of acetylcholinesterase. The Kyoto Encyclopedia of Genes and Genomes enriched differentially expressed genes and proteins are involved in signaling pathways including metabolic pathway, AD, pathway in cancer, PI3K-AKT signaling pathway, and cAMP signaling pathway. Transcriptomics and proteomics sequencing resulted in 19 differentially expressed genes and proteins. Finally, in contrast to the model group, after FN treatment, the protein expressions and genes associated with the PI3K-AKT pathway were significantly improved in RT-qPCR and Western blot and assays. This is consistent with the findings of transcriptomic and proteomic analyses. Our study found that, FN may improve some symptoms of AD model rats through PI3K-AKT signaling pathway.

Objective: To investigate the protective effects of pretreatment with a Shenfu(SF)injection on arrhythmias induced by myocardial ischemia-reperfusion(IR)injury in aged mice. Methods: Thirty 18-month-old C57BL/6J mice were randomly divided into 3 groups(n=10, each group): the sham treatment group(receiving sham operation without thoracotomy), the IR group(undergoing the ligation of the left anterior descending coronary artery with ischemia for 30 min and reperfusion for 2 h)and the SF group(receiving SF intraperitoneal injection of 10 ml/kg 30 min before thoracotomy and the same treatment as the IR group). Arrhythmias were monitored, and serum levels of creatine kinase-isoenzyme(CK-MB), troponin(cTnI)and tumor necrosis factor α(TNFα), and the ratio of myocardial connexin 43(Cx43)phosphorylation(ser368)to total protein(p-Cx43/t-Cx43)were detected in the three groups. Results: Ventricular arrhythmias occurred in the IR and SF groups.Compared with the IR group, ventricular arrhythmias in the SF group were alleviated, the frequency of ventricular premature systolic episodes was reduced(36.6±13.5 times vs. 48.4±22.1 times), the frequency of ventricular tachycardia/fibrillation decreased(3.4±1.8 times vs. 7.6±3.5 times), and the total duration of ventricular tachycardia/fibrillation episodes was shortened(8.9±4.5 times vs. 17.7±5.1 times)in the SF group(P<0.05), but there was no significant difference in arrhythmia scores(1.8±1.2 points vs. 1.9±1.7 points, P>0.05)between the two groups.Compared with the sham treatment group, serum levels of CK-MB, cTnI and myocardial TNF(increased in the IR and SF groups(P<0.05), and their levels were lower in the SF group than in the IR group(P<0.05). Compared with the sham treatment group, the ratio of Cx43 ser368/total protein was lower in the IR and SF groups, but was higher in the SF group than in the IR group(P<0.05). Conclusions SF pretreatment can significantly reduce IR-induced arrhythmias in aged mice possibly by reducing TNF(and up-regulating the phosphorylation activity of Cx43.

OBJECTIVE To prepare Gambogic acid （GA） nanocapsules （GA-LNCs） and Neogambogic acid （NGA） nanocapsules（NGA-LNCs），and to evaluate their antidiabetic activities. METHODS Using water as the aqueous phase ，medium- chain triglyceride as the oil phase and polyethylene glycol monostearate as the surfactant ，GA-LNCs and NGA-LNCs were prepared by phase inversion method. Using entrapment efficiency and drug-loading amount as index ，the formulation technologies of above 2 nanocapsules were optimized by simplex lattice design. Its physical and chemical properties were investigated. The diabetic mice model was established. GA-LNCs and NGA-LNCs （1.92 and 2.42 mg/kg respectively ）were given intragastrically ，once a day ，for consecutive 6 weeks. The fasting blood glucose of mice ，the activities of superoxide dismutase （SOD）and glutathione peroxidase （GSH-Px），the contents of malondialdehyde （MDA），total cholesterol （TC），triglyceride （TG），high-density lipoprotein cholesterol（HDL-C）and low-density lipoprotein cholesterol （LDL-C）were all detected. RESULTS The optimal formulation of 2 kinds of nanocapsules included 60％ water，10％ medium-chain triglyceride ，30％ polyethylene glycol monostearate （total amount of the three was 2 g）and 35 mg GA or NGA . The encapsulation efficiencies of GA-LNCs and NGA-LNC obtained by the optimal formulation were （92.01±0.68）％ and（93.12±2.11）％；the drug-loading amount were （0.99±0.21）％ and（1.21±0.22）％， respectively. GA-LNCs and NGA-LNCs were yellow ，homogeneous and transparent liquid without precipitation. They were spherical in microscopic shape ， and had obvious shell- Δ 基金项目：吉林省科技发展计划项目（No.20200404090YY）；吉 membrane structure. The particle sizes were （28.11 ± 9.76） 林省教育厅科学技术研究项目（No.JJKH20210372KJ） ＊硕士研究生 。研究方向 ：植物药 。E-mail：zhanhe0108@163. and（22.06±6.84）nm；Zeta potential were （－4.09±1.00） com and（－17.40±1.32）mV，and polydispersity were 0.93±0.06 # 通信作者：讲师，博士。研究方向：中药有效成分治疗疾病的作 and 0.74±0.12. The results of animal experiments showed that 用机制。E-mail：chenweijia_jlau@163.com both GA-LNCs and NGA-LNCs could sig nificantly increase 中国药房 2022年第33卷第9期 China Pharmacy 2022Vol. 33 No. 9 ·1075· the activities of SOD and GSH-Px and the seru m content of HDL-C （P＜0.05 or P＜0.01）in model mice ，and significantly decreased the fasting blood glucose and the serum contents of MDA ， TC， TG and LDL-C （P＜0.05 or P＜0.01）. CONCLUSIONS GA-LNCs，NGA-LNCs prepared in this study are good in physical and chemical properties and have good anti-diabetes activity.