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Objective To establish a sensitive,accurate and reliable LC-MS method to quantify the monoclonal antibody in rat blood and preclinical pharmacokinetics study. Methods The method of selective reaction monitoring(SRM)was established by using on-the-fly orthogonal array optimization(OAO)with anti-chemotherapeutic antibody-06(ACA-06)as a model drug,and the pharmacokinetics of ACA-06 in rats was studied. The optimal signature peptide selection and mass spectrometry conditions were opti?mized by single LC-MS run. Two highly specific and sensitive peptides,FSGVPDR and VNSAAFPAPIEK,were identified as the sig?nature peptides of quantification based on the OAO optimization result,which were located in the ACA-06 light chain and heavy chain complementary determining region(CDR)respectively. Results The method had good linearity in the range of 12.9~32250 ng/ml (r20.9819-0.9946). The lower limit of quantification was 12.9 ng/ml. The intra-day precision(RSD)was less than 11.5%,the inter-day RSD was less than 6%,and the accuracy was between-4.0%and 4.3%. Conclusion In this study,LC/SRM-MS method is es?tablished for the determination of ACA-06 in rat plasma. The method is specific,accurate and sensitive and enables an accurate phar?macokinetics investigation of ACA-06 in rat plasma.
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Objective To synthesize poly asparagine derivatives and to evaluate its safety at the cellular level, which provide research platform for its potential application as drug carrier. Methods Polysuccinimide was synthesized by ther?mal polymerization of L-polyaspartic acid, and the target product of PSI-Phe-EA was obtained by the ring-opening reaction of polysuccinimide using L-phenylalanine methyl ester hydrochloride and ethanol amine. The structure of PSI-Phe-EA were characterized by 1H NMR. The rate of ring-opening of PSI was calculated by internal standard method of 1H NMR. The change of hydrophilicity was studied by the comparison of solubility. The cytotoxicity and morphology modification by PSI-Phe-EA at designate concentrations was investigated by MTT method and inverted microscopy respectively. The effects on cell cycles were analyzed by flow cytometry after propidium iodide (PI) staining. Results 1H NMR results confirmed the structure of PSI-Phe-EA and the ring-openning rate of PSI was 40%. The hydrophilicity of PSI-Phe-EA was greatly in?creased upon ring opening using ethanol amine. MTT test showed that the cell survival rates of NIH 3T3 and HepG2 cells were higher than 80%under the examined concentration (<100 mg/L). Inverted microscopy showed that 50 mg/L of PSI-Phe-EA treatment had no adverse effects on cell morphology. Cell cycle analysis indicated that PSI-Phe-EA treatment had no in?fluence on cell cycles of NIH 3T3 and HepG2 cell lines. Conclusion PSI-Phe-EA showed high hydrophilicity without sig?nificant effects on the cells survival, cells morphology and cell cycles. It is a kind of safe polymer material.
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Objective To study the correlations of pelvic endometriosis cysts with pain and infertility according to the revised American Fertility Society (r-AFS) staging of endometriosis.Methods We retrospectively studied the clinical information of 258 cases patients with the ovarian endometriosis cysts who undergoing laparoscopic ovarian endometriosis cysts excision from January 2007 to December 2013.In accordance with the r-AFS,the correlation between pelvic adhesions and pain symptoms or infertility was evaluated.Results Among 258 cases with ovarian endometriosis cysts,there were 72.09％ (186/258) of patients presented pelvic pain and 16.28％ of patients presented infertility.The stages of r-AFS were that 16 cases in stage I,80 cases in stage Ⅱ,108 cases of in stage Ⅲ and 54 cases of stage Ⅳ.Also,186 cases with pain symptom,77.42％ (144/186) of patients were in stage Ⅲ and Ⅳ,which was remarkably higher than that of no symptom group (25.00％ (18/72),x2 =64.017,P ＜ 0.05).Among 42 cases with infertility,there were 85.71％ (36/42) of patients were in stage Ⅲ and Ⅳ,remarkably higher than of no infertility group (58.33％ (126/216),x2 =11.283,P ＜ 0.05).Conclusion The r-AFS stage of pelvic distribution in ovarian endometriosis cyst is closely correlated with pain symptoms and infertility.Patients in stage Ⅲ and Ⅳ with pain and infertility are more significant.