OBJECTIVE:To determinate the serum concentration of methotrexate by HPLC. METHODS:The samples were separated on a Waters C18 column at a column temperature of 30 ℃. The mobile phase consisted of acetonitrile-phosphate buffer(pH 7.2,12∶88) at a flow rate of 1.0 mL?min-1. The detection wavelength was set at 303 nm. RESULTS:The linear range of methotrexate was 0.5～80 ?g?mL-1(r=0.999 4). The methodological recovery rates were all above 90%; the intraday RSD was 3.03 %～3.52 % and the inter-day RSD was 2.57%～4.05%. CONCLUSION:The method is proved to be simple,rapid,accurate and sensitive,and it is suitable for the determination of serum concentration and pharmacokinetic study of methotrexate.

Objective:To improve the affinity of an anti TNF? scFv.Methods:Starting from an anti TNF? scFv gene a mutant phage antibody library was generated by error prone PCR.Affinity improved clones were selected and subjected to staggered extension process to shuffle the mutated sites.Mutants with further improved affinity were selected by bio panning.Affinity was judged by dot blot ELISA and thiocyanate elusion ELISA.Results:Seven affinity improved mutants were obtained from library constructed by error prone PCR.By StEP mediated shuffling of these 7 clones and via bio panning,mutants with further improved affinity were obtained.Conclusion:Combination of error prone PCR and StEP could be used to improve the affinity of antibodies. [

OBJECTIVE: To observe the effect of electronic moxibustion on memory function in the patients with amnestic mild cognitive impairment (aMCI). METHODS: A total of 59 aMCI patients were randomized into an electronic moxibustion group (30 cases) and a placebo moxibustion group (29 cases). In the electronic moxibustion group, the electronic moxibustion was applied to Baihui (GV 20), Dazhui (GV 14), Mingmen (GV 4) and Taixi (KI 3), 45 ℃ in temperature, 20 min each time. The treatment was given once a day, 5 times a week. The treatment for 4 weeks was as one course and 2 courses were required totally. In the placebo moxibustion group, the moxa-free patch was used, 38 ℃ in temperature. The acupoint selection and the treatment frequency were same as the electronic moxibustion group. Before and after treatment, Rivermead behavior memory test (RBMT) was adopted to evaluate the global memory function of the patients in the two groups and the N-back task test was adopted to evaluate working memory function separately. Additionally, the mini-mental state examination (MMSE) and its immediate memory, Montreal cognitive assessment (MoCA) and its delay recall were adopted to evaluate the global cognitive function and memory function RESULTS: In the electronic moxibustion group, after treatment, RBMT score, N-back accuracy rates, MMSE and MoCA scores and the scores of immediate memory and delay recall were improved significantly as compared with those before treatment (<0.01). In the placebo moxibustion group, the accuracy rates of 1-back and 2-back task and the scores of immediate memory and delay recall were improved obviously as compared with those before treatment (<0.05, <0.01). After treatment, the improvements of RBMT score, the accuracy rates of N-back task and MMSE and MoCA scores in the electronic moxibustion group were higher than those in the placebo moxibustion group (<0.05). CONCLUSION Electronic moxibustion improves memory function in the patients with amnestic mild cognitive impairment.
Acupuncture Points ; Amnesia ; therapy ; Cognitive Dysfunction ; therapy ; Humans ; Memory ; Mental Status and Dementia Tests ; Moxibustion ; methods