1.Expression of EGFR mutation-specific antibody and its significance in lung adenocarcinoma
Xia GU ; Jieyu WU ; Xinming HE ; Yunen LIN ; Ping HE ; Qinian WU ; Guangqiu LI
Chinese Journal of Clinical and Experimental Pathology 2015;(6):652-656
Purpose To investigate the expression and significance of epiderma1 growth factor receptor( EGFR)mutation-specific anti-bodies in 1ung adenocarcinoma. Methods Immunohistochemica1( IHC)technique was used to detect the expression of EGFR muta-tion-specific antibodies(EGFR-19,EGFR-21)and EGFR tota1 protein antibody(EGFR-P)in 171 cases of 1ung adenocarcinoma with resected specimens,and EGFR gene mutation was a1so performed by amp1ification refractory mutation aystem-PCR( ARMS-PCR). The expression of EGFR-19,EGFR-21 and EGFR-P mutant proteins was compared with EGFR gene mutation and their re1ationship with histo1ogica1 c1assification and c1inica1 characteristics were ana1yzed. Results The expression of EGFR mutant protein was corre1ated with the poor differentiation group inc1uding micropapi11ary and so1id predominant types( P=0. 021 ). EGFR-21 high expression was re1ated to p1eura1 invasion(P=0. 005). The coherence of IHC(with EGFR-19/21 antibodies)and ARMS-PCR was existed(Kappa>0. 4 ). Taking ARMS-PCR as a standard,the sensitivity and specificity of EGFR-19 and EGFR-21 were 65. 0% and 89. 4%, 70. 0% and 97. 6%,respective1y. Conclusions Expression of EGFR mutation-specific antibodies is associated with poor differentia-tion and p1eura1 invasion. It suggests a worse prognosis indicator in 1ung adenocarcinoma. Because of the coherence with ARMS-PCR, using IHC with mutation-specific antibodies to detect the mutant proteins of EGFR-19/21 may act as a pre1iminary screening method of EGFR gene mutation.
2.Study on the fingerprint and content determination of Xiaohe syrup
Na LI ; Xiaoxiao WANG ; Jieyu XIA ; Yu LIU ; Jianling DENG ; Wanyi CHEN
China Pharmacy 2024;35(12):1457-1462
OBJECTIVE To establish high performance liquid chromatography (HPLC) fingerprint of Xiaohe syrup and determine the contents of 10 effective ingredients in them. METHODS With 12 batches of Xiaohe syrup as samples, ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was adopted with Athena C18 (250 mm×4.6 mm, 5 μm) as the chromatographic column, acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL/min, and the detection wavelength was 210 nm. Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprint (2012A version) was imported to establish the fingerprint of Xiaohe syrup and evaluate the similarity. The content determination was performed on ACQUITY UPLC BEH C18( 100 mm×2.1 mm, 1.7 μm) chromatographic column, with 0.01% formic acid acetonitrile-0.01% formic acid water as mobile phase for gradient elution at a flow rate of 0.4 mL/min; combined with high-resolution mass spectrometer, positive and negative ions were scanned with an electric spray ion source to determine the content of each main component in 12 batches of Xiaohe syrup. RESULTS A total of 33 common peaks were calibrated in 12 batches of samples, with similarities greater than 0.97; 10 chromatographic peaks were confirmed, namely flavonoid glycosides, paeoniflorin, ferulic acid, naringin, rosmarinic acid, neohesperidin, salvianolic acid B, tetrahydropalmatine, saikosaponin A, and saikosaponin D. The results of content determination showed that the above 10 components had good linear relationships within their respective mass concentration ranges (all R 2>0.999), with contents ranging from 0.35 to 0.64, 3.15 to 5.61, 0.11 to 0.17, 1.68 to 3.17, 1.59 to 1.90, 1.15 to 1.64, 0.78 to 1.48, 0.11 to 0.26, 0.06 to 0.13, and 0.33 to 0.61 mg/mL, respectively. CONCLUSIONS The main components of 12 batches of Xiaohe syrup are similar, but the contents vary; HPLC fingerprint and UPLC-MS/MS content determination method established in this study can be used for comprehensive quality evaluation of Xiaohe syrup.