1.Correlation of HPV types with Th17 and Treg cells in cervical cancer
Donghui CAI ; Yan LI ; Runmei GUO ; Tingting ZHANG ; Zhifang CHEN
Chinese Journal of Clinical Oncology 2016;43(24):1099-1102
Objective:Based on the detection of CD4+IL17+Th17 and CD4+CD25+Foxp3+Treg cells in the peripheral blood of patients with human papilloma virus (HPV), the study analyzes the ratio of Th17 to Treg and compares the different expression of cytokine IL-17, IL-10, TGF-β, and IL-23 in the serum. Methods:From September 2014 to June 2016, 119 cases of untreated patients with cervical HPV in-fection were collected at the First Affiliated Hospital of Xinjiang Medical University, which include 46 cases of cervical cancer (CC), 43 cases with cervical intraepithelial neoplasia (CIN)Ⅱ-Ⅲ, and 30 with chronic cervicitis (CCS). The detection of HPV types classified 80 patients as high-risk and 39 as low-risk. Flow cytometry examined Treg and Th17 cells, and enzyme-linked immune sorbent assay was used to test related cytokines. Results:In the infection of high-risk cervical HPV, the rate of the CC group was 87%(40/46), which was higher than that of the CINⅡ-Ⅲgroup and the CCS group, which were 77%(33/43) and 23%(7/30), respectively. For the expression of Th17 and Treg cells in the peripheral blood and the related cytokine IL-17, IL-23, TGF-β, and IL-10 in the serum, the levels of CC group were also significantly higher than that of the other two groups, and the ratio of Th17 to Treg increased considerably with the se-verity of cervical lesions. The expression of Th17 and Treg cells in the peripheral blood of patients with high-risk cervical HPV were higher than that of patients with low-risk cervical HPV (P<0.05), which was the same for the expression of the related cytokine IL-17, IL-23, TGF-β, and IL-10 in the serum. The ratio of the Th17 to Treg of patients with high-risk cervical HPV increased remarkably (P<0.05). Conclusion:The expression of Treg and Th17 cells in high-risk patients exhibits an increasing trend, and the ratio of increased Th17 to Treg may directly participate in the process of HPV infection and immune evasion, which plays an important role in the occurrence and development of cervical cancer.
2.Experimental methodology of simultaneous determination of carbamazepine, phenytoin, and phenobarbital in serum by high-performance liquid chromatography
Runmei XIAO ; Zhenli GUO ; Jingzhi SUN ; Ruijie LIN ; Zhaohui HUANG ; Yong CHEN
Journal of Chinese Physician 2013;15(10):1322-1326
Objective To establish a high-performance liquid chromatography (HPLC) method with diode array detection to simultaneously determine carbamazepine,phenytoin,and phenobarbital in serum.Methods Extraction solvent (800μl ethylene acetate) and sample (0.2 ml) was mixed,extracted for 2 min,and centrifuged (3500 r/min,4 minutes).A volume (600 μl) of extract liquor was volatilized to dryness in water bath with the volatilization temperature 75 ℃,then was redissolved with 1.0 ml mobile phase.Analysis conditions was column temperature 30°,mobile phase (methanol∶ water =40∶60),and detection wavelength of 254 nm.Three metabolites were effectively separated.Results Under the optimized condition,calibration curves of three metabolites were linear in the ranges of (1.52 ~ 120 mg/L) and the correlation coefficients were not less than 0.999.The detection limits (S/N =3) were in the range of 0.4 ~ 1.5 mg/L.The spiked recoveries were in the range of 91.3% ~ 111% with relative standard deviations (RSD) less than 5%.Conclusions The optimal pretreatment condition for the sample was established.The chromatographic separation and the detection condition were optimized.The method was sensitive and accurate,and could meet the need of monitoring serum drug concentration.