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Objective:Through gene cloning,expression and activity analysis of hBcl 2 to provide enough proteins for the development of new drugs on the basis of 3 D structue of hBcl 2 protein.Methods:Gene cloning using PCR amplification,identification with Western Blot and MALDI TOF MS.Results:hBcl 2 gene was cloned,inserted into pET28a(+) and solublely expressed in E.Coli.Its molecule weight was confirmed through MALDI TOF MS,which fits exactly with its theoretical value.After purification to reach electrophoresis homogeneity,it showed the ability of combining specifically with BH 3 domain of Bak,and then provide the basis for the further research on small chemical compounds which could specifically bind hBcl 2 protein.Conclusion:In this work hBcl 2 was successfully expressed and recovered its binding activity in a soluble form. [
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Huichunzhibao oral liquid (HZOL ) is a traditional Chinese herb Preparation composedof Panax ginseng, Hairy Antler (Cervus nippon Tcmminck ) and Epimedium brevicornum Maixm..The active principle of each component was identified by TLC and icariin, the main active principle of E. brevicornum was determined quantitatively by HPLC. The meth od was found to be accuratc, sensitive and reproduciblc with average recovery 98.97 % andRSD = 1.53 (n = 3).
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Objective:To establish the determination of paeoniflorin in Gubenyichang Tablet.Methods:RP-HPLC method was used for determination of paeoniflorin in Gubenyichang Tablet on C 18 column. The mobile phase was consisted of methanol-water (32∶68) and detection wavelength was at 230nm.Results:The average recovery was 98.9% and RSD was 1.22%. Conclusion:The method is simple, accurate and with good speciality.
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Objective: To determine cinnamaldehyde and cinnamic acid in Ramulus Cinnamomi and Guizhifuling Capsules.Methods: RP HPLC method was used. The samples were separated on a Zobax ODS column with the mobile phase of acetonitrile:0.1% phosphoric acid (34∶76) and the detection wavelength was set at 285nm.Results: The calibration curves were linear in the range of 3.34～53.4?g?mL -1 in cinnamaldehyde ( r= 0.9996) and 0.804～12.9?g?mL -1 in cinnamic acid ( r =0.9999),respectively. The average recoveries of cinnamaldehyde in Ramulus cinnamomi and Guizhifuling capsules were 98.6% ( RSD =1.35% n=6 ) and 97.4% ( RSD =1.62% n =6), respectively and of cinnamic acid were 99.2% ( RSD =0.97% n=6 )and 100.0% (RSD =0.73%, n=6 ),respectively.Conclusion: The method is quick, acurate and suitable for the determination of Ramulus Cinnamomi and its compound Chinese medicinal preparations.