1.Optimization of Extraction Process for Arctiin and Arctigenin inArctium lappaL. Based on Central Composite Design and Response Surface Methodology
Enbo CAI ; Wei WANG ; Caixia JIA ; Yan ZHAO ; Dalong WANG ; Xiangxiang LIU ; Dingrong YE ; Limin YANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(9):1828-1832
This study was aimed to optimize the extraction process of double-marker components for Arctium lappa L. The central composite design and response surface methodology was used. According to 3 main factors, the extraction rates of arctiin and arctigenin was used as evaluation indexes. Multiple linear regression and two-order polynomial equation were used. The binomial fitting model was performed in the optimization of arctiin and arctigenin extraction technology. The results showed that the indentified optimized extraction technology of arctiin and arctigenin was 70% ethanol, 24-fold, ultrasonic solvent extraction for 15 minutes. It was concluded that this technology was able to extract large amount of arctiin and arctigenin, which provided experiment evidences for arctiin and arctigenin preparation. It also provided references for the development and utilization of arctiin and arctigenin.
2.Value of diagnosis and efficacy evaluation of folate receptor-positive circulating tumor cells in patients with small cell lung cancer
Xianghua LU ; Huang CHEN ; Fei SU ; Ruiying JIANG ; Ye WANG ; Dingrong ZHONG ; Liya LI
Cancer Research and Clinic 2021;33(1):33-37
Objective:To analyze the value of folate receptor-positive circulating tumor cells (FR +-CTC) in the diagnosis and efficacy evaluation of patients with small cell lung cancer (SCLC). Methods:The data of 59 patients with SCLC and 14 patients with benign pulmonary diseases treated in China-Japan Friendship Hospital from May 2017 to October 2019 were retrospectively analyzed. Folate receptor targeted detection was used to detect the level of FR +-CTC in the blood of SCLC patients. The levels of serum progastrin-releasing peptide (Pro-GRP), neuron-specific enolase (NSE), cytokeratin 19 fragment 21-1 (Cyfra21-1) , and carcinoembryonic antigen (CEA) were detected by using chemiluminescence. The median ( P25, P75) was used as all the detection indexes. Mann-Whitney U test was used for pairwise comparison, Spearman correlation test was used to analyze the correlation between two variables, and receiver operator characteristic (ROC) curve was used to evaluate the diagnostic efficacy. Results:The level of FR +-CTC in 59 patients with SCLC was 11.00 FU/3 ml (7.10 FU/3 ml, 14.50 FU/3 ml), and the positive rate of FR +-CTC in patients with SCLC was 66.10% (30/59); the level of FR +-CTC in 14 patients with benign pulmonary diseases was 6.75 FU/3 ml (5.03 FU/3 ml, 7.85 FU/3 ml), and the positive rate of FR +-CTC in 14 patients with benign pulmonary diseases was 14.29% (2/14). The level of FR +-CTC in patients with SCLC was higher than that in patients with benign pulmonary diseases, and the difference was statistically different ( U = 33.50, P < 0.01). The expression level of FR +-CTC was not related to age, gender and smoking history in SCLC patients (all P>0.05). The expression level of FR +-CTC in patients with extensive-stage was higher than that in patients with limited-stage, and the difference was statistically significant ( P < 0.05). Tumor markers Pro-GRP, NSE, Cyfra21-1 and CEA were compared with FR +-CTC, and the ROC curve was drawn; the results showed that FR +-CTC had better sensitivity (71.2%) and specificity (92.90%) in the diagnosis of SCLC. For SCLC patients who received chemotherapy, the decrease range of FR +-CTC in patients with partial remission and stable disease was greater than that in patients with the progression of disease, and the differences were statistically significant (all P < 0.05). Conclusion:FR +-CTC can assist the diagnosis and disease staging of SCLC. For patients receiving chemotherapy, continuous detection of circulating tumor cells can help to evaluate the efficacy of chemotherapy and provide a reference for the choice of clinical treatment.
3. Interpretation of BRAF gene mutation detection by real-time PCR: a study of 8 644 thyroid fine-needle aspiration specimens
Ye WANG ; Jiping DA ; Lei YANG ; Huang CHEN ; Dingrong ZHONG
Chinese Journal of Pathology 2019;48(11):873-877
Objective:
To study common problems in BRAF gene mutation detection, and conditions for repetition testing using thyroid fine needle aspiration specimens.
Methods:
A total of 8 644 cases of thyroid fine-needle aspiration specimens at China-Japan Friendship Hospital were collected between February, 2012 and July, 2018. BRAF gene mutation was detected by real-time PCR. Repeat testing was performed in 237 cases when the results were inconsistent with clinical or cytological diagnosis or when uncertain results were obtained.
Results:
The final positive rates of BRAF mutation was 22.0% (1 897/8 625). Nineteen cases were excluded due to inadequate DNA samples. The average Ct value of internal quality control was 16.061, and the average Ct value of the positive samples was 19.147. Among 237 repeat tests, 51.4% (19/37) continued to have poor DNA quality and 48.6% (18/37) had adequate DNA resulting in 1 positive case and 17 negative cases. In 40 repetition of initial negative cases, results were unchanged. In initial positive cases, 40.4% (40/99) with a difference of Ct value (between BRAF gene and internal quality control) between 8 to 12 turned negative after repetition, 69.8% (37/53) of these cases with a difference of more than 12 turned negative after repetition. The sensitivity and specificity of BRAF mutation were 83.97% and 96.94%, respectively.
Conclusions
Difference between BRAF gene Ct value and internal quality control Ct value is recommended as a reliability index for the test result. Cases with a difference greater than 8 should be subjected to repeat testing.