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.

OBJECTIVE:To establish the method for the determination of oleanolic acid and ursolic acid in different medicinal parts of Tibetan medicine Pteocephalus hookeri,and compare the differences among the different parts. METHODS:The contents of oleanolic acid and ursolic acid from different medicinal parts(whole plant,aerial part,underground part)of P. hookeri were de-termined by UPLC-PDA. The separation was performed on Acquity UPLC HSS T3 column(150 mm×2.1 mm,1.8 μm)with mobile phase consisted of methanol-0.1 mol/L ammonium acetate(88:12,V/V)at the flow rate of 0.2 mL/min. The detection wavelength was set at 210 nm,and column temperature was 30 ℃. The sample size was 5 μL. RESULTS:The linear ranges of oleanolic acid and ursolic acid were 10.65-1065 μg/mL (r=0.9996) and 18.8-1880 μg/mL (r=0.9994),separately. The recoveries were 96.95%(RSD=1.24%,n=9) and 98.12%(RSD=2.13%,n=9),separately. RSDs of precision,stability and reproducibility tests were all less than 3%. The contents of oleanolic acid and ursolic acid from different medicinal parts in P. hookeri were in de-scending order of aerial part>whole plant>underground part;the average total content of oleanolic acid and ursolic acid from whole plants was 0.35%,the aerial part reached 0.56% and underground part was 0.09%. CONCLUSIONS:The method is rapid, accurate and reproducible,and it is suitable for the content determination of oleanolic acid and ursolic acid in different medicinal parts of Tibetan medicine P. hookeri. The contents of oleanolic acid and ursolic acid from aerial part of P. hookeri are higher than whole plant and underground part. It is suggested to use aerial parts of medicine.

Objective To prepare human alpha segment of high affinity IgE receptor(FcεRIα)protein by genetic engineering technology and to identify its biological function for laying the foundation for further researching the role of FcεRIα in allergic disea-ses.Methods The human FcεRIα gene was obtained by the PCR based accurate synthesis(PAS)method and the prokaryotic ex-pression vector pET-28a(+)was constructed.The FcεRIα was expressed at low temperature induction and the recombinant protein was purified by His tag.The biological function of recombinant human FcεRIα protein was identified by ELISA.Results The hu-man FcεRIα gene was amplified by PAS with a size of approximately 560 bp.The pET-FcεRIα plasmid was correct through the double enzyme digestion and sequencing identification.The human FcεRIα with a molecular weight of approximately 22 000 was in-duced and purified.The recombinant human FcεRIα could effectively detect human serum anti-FcεRIα autoantibody and could com-bined with serum IgE antibodies with high efficiency.Conclusion Human FcεRIα protein is successfully prepared,which prelimina-rily has the ability for detecting the human serum anti-FcεRIα autoantibodies and IgE antibodies,and provides a favorable practical base for further study.

[Abstract] [Objective] To establish a new centrifugal-membrane hybrid plasmapheresis (CMHP) model, and observe its clinical efficacy, safety and advantages in the treatment of hyperlipidemia by comparing with centrifugation therapeutic plasma exchange (cTPE). [Methods] A retrospective analysis was performed on 47 patients with hyperlipidemia treated in our department from August 2021 to September 2023, with a total of 60 treatments. They were divided into two groups: CMHP group with 37 patients and 46 treatments; cTPE group included 10 patients with 14 treatments. In the CMHP group, the plasma was separated by a blood cell separator and the plasma components were separated by a two-stage membrane plasma component separator. In the cTPE group, the plasma was separated by a blood cell separator for therapeutic plasma exchange. The clinical efficacy, safety and advantages of the two groups were compared. [Results] The reduction ratios of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) before and after treatment in CMHP group were 68.56%[59.81%, 73.42%], 65.80%[55.55%, 75.98%] and 46.57%[36.02%, 54.83%] and 66.67%[43.48%, 76.24%] respectively, with statistically significant difference (P<0.001). The decrease ratios of TC, TG and HDL-C before and after treatment in cTPE group were 42.52%[29.67%, 49.85%], 52.32%[38.43%, 67.07%] and 22.36%[8.51%, 33.65%], respectively, and the difference was statistically significant (P<0.05). The plasma treatment multiple was 1.48 (0.29) in CMHP group, which was significantly higher than 0.87 (0.26) in cTPE group, with a statistically significant difference (P<0.001), resulting in higher TC and HDL-C reduction ratios in CMHP group than in cTPE group, with a statistically significant difference (P<0.001), while there was no significant difference in TG reduction rate between the two groups (P>0.05). At the same time, the cTPE group required 1 520.00 mL[1 462.50 mL, 2 000.00 mL] plasma input, while the CMHP group achieved zero blood input. The adverse reaction ratio was 6.52%(3/46) in CMHP group and 7.14%(1/14) in cTPE group, and the difference was not statistically significant (P>0.05). [Conclusion] Compared with cTPE, CMHP can better reduce blood lipid levels without any blood products, avoid the spread of blood infectious diseases, and have a low incidence of adverse reactions, so it has a good clinical application prospect.