Oleanolic acid (OA) and ursolic acid (UA) are isomeric triterpenic acids and only one methyl's position is different between them.OA and UA always exist in the same plant,so it is difficult to separate them when determining contents by RP-HPLC.In this study,a very simple mobile phase for HPLC was developed to simultaneously determine UA and OA,and the factors affecting separation were also discussed.The mobile phase is methanol:water (95∶5) with flow rate 0.4mL/min.The retention time for OA and UA was 20.58 and 21.57 min,respectively,the resolution was 1.61.The average contents of OA and UA of three Loquat leaves sets were 1.4 mg/g and 5.6 mg/g,respectively.Regarding the HPLC,we found that changing mobile phase,adjusting the pH value or adding ion-pairing agent could not affect the separation between UA and OA greatly.While adjustment of the flow rate and column temperature could improve the resolution greatly.

Description of the tasks provided by the hospital to Beijing Olympic Games and their characteristics, and a summary of the practice and experiences in the course. For example, a general view of hospital mission, reasonable tasks scheduling, and handling of the four relationships. It holds that medical institutions need to take the opportunity of providing medical services to large international events to upgrade and improve themselves, standardize their management, reinforce normalization in medical practice, and better their brand marketing awareness.

Objective To investigate the application of the central venous catheter (CVC) specialized for drainage intervention in severe acute pancreatitis (SAP) patients.Methods Sixty-two severe acute pancreatitis patients with seroperitoneum were randomly (random number) assigned into two groups:the drainage group (n =31) and the control group (n =31).All patients were treated with conventional internal medicine therapy.Patients of drainage group were treated with continuous peritoneal drainage by using the central venous catheter.The intra-abdominal pressure (IAP),lactic acid (LAC),and procalcitonin (PCT) were detected before and 12 h,24 h,48 h,72 h,5 days after intraperitoneal drainage.The symptoms of abdomen pain,abdomen distention,resume of bowel movement and the rate of MODS were observed.Results All patients with drainage got catheter successfully inserted.Compared with the control group,the IAP,LAC and PCT decreased significantly in the patients of drainage group.And the duration of abdomen pain,abdomen distention and resume of bowel movement function in the drainage group were shorter and the rate of MODS was lower.Conclusions Application of CVC specialized for intraperitoneal drainage is a safe and effective method for the treatment of pancreatitis with seroperitoneum.It is worthwhile to be widely used in clinic.

Objective To clone and express the partial encoding sequence of Mr 70 000 heat shock protein of Cryptosporidium andersoni (CaHSP70) in Escherichia coli and identify the recombinant protein. Methods Total RNA was extracted from oocysts of C.andersoni isolated from Xuzhou, Jiangsu (XZ-BOV). The CaHSP70 gene was amplified by RT-PCR. The PCR product was cloned and then subcloned into pET28a vector, and the recombinant plasmids were transformed into E.coli BL21(DE3) subsequently. The expressed protein induced by IPTG was purified and identified by SDS-PAGE and Western blotting, and was further analyzed by relevant bioinformatics softwares. The specific IgG antibodies in mice immunized by rCaHSP70 were detected by Western blotting and ELISA respectively. Results The deduced amino acid sequence showed to be identical with that of C. andersoni Mr 70 000 heat shock protein (HSP70). The recombinant protein expressed in the form of inclusion body was about Mr 43 000. It could be recognized by anti-His G labeled HRP antibodies and all the sera from mice infected with C. andersoni and children infected with C. parvum as well as sera from mice immunized with rCaHSP70 respectively. The rCaHSP70 possibly had multiple domains and potential antigenic determinants. Phylogenetic analysis showed that XZ-BOV and C. andersoni were in the same clade. ELISA showed that the level of specific antibodies against rCaHSP70 in immunized BALB/c and C57BL/6 mice was significantly higher than that of mice before immunization. Conclusion The recombinant plasmid pET28a-CaHSP70 has been constructed. The purified rCaHSP70 exhibits high antigenicity and seems a potential candidate antigen for immunodiagnosis of cryptosporidiosis.

AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thy-roid follicular epithelial ( FRT) cells and its electrophysiologic properties .METHODS: The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection .In order to ob-tain the high efficiency of gene transfection and expression , the quantity and ratio of lipid/DNA complexes were optimized . The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence .The expression and location of TMEM 16A in the FRT cells were observed under an in-verted fluorescence microscope .TMEM16A protein was associated with calcium-dependent chloride current , as measured with halide-sensitive fluorescent protein and patch-clamp technique .RESULTS: The results of double digestion and se-quencing indicated that TMEM16A was cloned into pUB6/V5.The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A.The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM 16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L.CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed.TMEM16A is the molecular identity of calcium-activated chloride channels .

Objective This study aimed to explore issues about dysandia female during pregnancy,peripartum and contraception and to supply reference for their reproductive health improvement. Methods Purposive sampling strategy was used. With the aid of an interview guide, 12 dysaudia women who met inclu-sion and exclusion criteria were invited to be interviewed. Interviews were recorded, validated and then ana-lyzed. Results Four themes were generated, limited knowledge, passive information acquisition, total depen-dence on family and reluctant to talk about contraception. Total dependence on family was noticeable during pregnancy and peripartum. Though little was known about health protection related to pregnancy, peripartum and contraception, participants were passive at seeking for relevant information. Conclusions Deaf women may experience more threats to reproductive health in relation to pregnancy, peripartum and contraception that they require more guidance and assistance.