Detection of DNA methylation of HYAL2 gene for differentiating malignant from benign thyroid tumors / 南方医科大学学报

OBJECTIVE@#To assess the value of DNA methylation level of HYAL2 gene as a molecular marker for differential diagnosis of malignant and benign thyroid tumors.@*METHODS@#DNA methylation of HYAL2 gene in tissue specimens of 190 patients with papillary thyroid cancer (PTC) and 190 age- and gender-matched patients with benign thyroid tumors was examined by mass spectrometry, and the protein expression of HYAL2 was detected immunohistochemically for another 55 pairs of patients. Logistic regression analysis was performed to calculate the odds ratio (OR) and evaluate the correlation of per 10% reduction in DNA methylation with PTC. Receiver operating characteristic (ROC) curve analysis was performed and the area under curve (AUC) was calculated to assess the predictive value of alterations in HYAL2 methylation.@*RESULTS@#Hypomethylation of HYAL2_CpG_3 was significantly correlated with early-stage PTC (OR=1.51, P=0.001), even in stage I cancer (OR=1.42, P=0.007). Age-stratified analysis revealed a significantly stronger correlation between increased HYAL2_CpG_ 3 methylation and early-stage PTC in patients below 50 years than in those older than 50 years (OR: 1.89 vs 1.37, P < 0.05); ROC analysis also showed a larger AUC of 0.787 in younger patients. The results of immunohistochemistry showed that patients with PTC had significantly higher protein expressions of HYAL2 than patients with benign tumors.@*CONCLUSION@#The alterations of DNA methylation level of HYAL2 gene is significantly correlated with early-stage PTC, suggesting the value of DNA methylation level as a potential biomarker for differentiation of malignant from benign thyroid tumors.

Effect of Different Hot Air Drying Methods on Chemical Constituents of Trichosanthis Fructus / 中国实验方剂学杂志

Objective:To compare the effects of different drying methods on the chemical constituents of Trichosanthis Fructus. Method:Trichosanthis Fructus was dried by means of air drying， sun drying， hot air drying （40， 60， 80 ℃） and variable temperature drying （50-80， 80-50 ℃）. The contents of nucleosides and flavonoids in Trichosanthis Fructus peels and seeds treated by different methods were compared by high performance liquid chromatography （HPLC）， mobile phase was acetonitrile-0.2% acetic acid aqueous solution （3∶7） （A）-acetonitrile （B） for gradient elution （0-15 min， 97-95%B； 15-30 min， 95%-90%B； 30-35 min， 90%-87%B； 35-40 min， 87%-86.5%B； 40-48 min， 86.5%-97%B； 48-50 min， 97%B）， the detection wavelength was 260 nm， and the flow rate was 0.4 mL·min^-1. Gas chromatography-ion mobility spectrometry （GC-IMS） was used to compare the changes of volatile components in the samples treated by different treatments. The volatile components were incubated on a SE-54 capillary column （0.32 mm×30 m， 0.25 μm） at 80 ℃ and 500 r·min^-1 for 15 min， the injection temperature was 85 ℃， the injection volume was 400 μL， the analysis time was 35 min， carrier gas was high purity nitrogen， the flow rate of carrier gas was 2.0 mL·min^-1， the flow rate of drift gas was 150 mL·min^-1， and the temperature of IMS detector was 45 ℃. Result:The contents of uridine， adenosine and adenine were higher after hot air drying at >50 ℃. Low temperature drying was conducive to maintaining the stability of cytidine， cytosine， rutin， luteolin and 2ʹ-deoxyadenosine. GC-IMS technology could realize the analysis and identification of Trichosanthis Fructus samples after different treatments. There were more volatile components after hot air drying at 80 ℃ and variable temperature drying. Conclusion:Hot air drying at 40 ℃ and 60 ℃ can retain nucleosides and flavonoids， and the volatile components are similar to those in traditional drying methods， which has the advantages of high efficient， controllable and suitable for industrial production.

Downregulation of LINC01508 contributes to cisplatin resistance in ovarian cancer via the regulation of the Hippo-YAP pathway / 부인종양

Background@#Some long non-coding RNAs (lncRNAs) have been found to contribute to cisplatin resistance. Here, we identified a novel lncRNA that was downregulated in cisplatin-resistant to ovarian cancer (OC) cells and aimed to examine the contribution of LINC01508 to cisplatin resistance in OC cells. @*Methods@#Differences in the lncRNA expression profile between OV2008 and C13K cells were assessed by lncRNA expression microarray. The expression of LINC01508 in ovarian epithelial cells, four OC cells, and OC, benign ovary tumor and normal ovary, cisplatin-resistant and non-resistant OC specimens were evaluated by quantitative real-time polymerase chain reaction (qPCR). The role of LINC01508 in OC cisplatin-resistant was evaluated by cell counting kit-8 (CCK-8), flow cytometry, colony formation, wound healing, Transwell, and tumor growth inhibition study in vivo. The clinical associations of LINC01508 in OC were evaluated using correlation analysis. The effects of verteporfin (VP) on cisplatin were explored to reveal the function of the hippo-YAP pathway on the cisplatin tolerance of C13K. @*Results@#LINC01508 was downregulated in cisplatin-resistant OC cells and platinum-resistant OC tissue (p<0.01). LINC01508 downregulation was correlated with tumor size, residual tumor, and platinum resistance. The overexpression of LINC01508 improves in vitro and in vivo sensitivity to cisplatin while predicts the poor overall survival which need further follow-up research. The increased level of LINC01508 could suppress the cisplatin resistance of OC cells through the inhibition of the hippo-YAP pathway. @*Conclusions@#The study proposes that dysregulation of LINC01508 expression results in resistance of OC to cisplatin through the inhibition of the hippo-YAP pathway.

Rapid discovery of antioxidants in Salvia miltiorrhiza by UPLC-DPPH- PDA-ESI-TOF/MS online technique / 中草药

Objective: To establish a method for rapid screening and identifying antioxidants in natural products by ultra high performance liquid chromatography-(1,1-dipheny l-2-trinitrophenylhydrazine)-photo diode array-electrospray-time-of-flight mass spectrometry (UPLC-DPPH-PDA-ESI-TOF/MS) and apply it to the rapid discovery of antioxidants from Salviamiltiorrhiza extracts. Methods: The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC HSS T3 column (100 mm×3.0mm,1.8 μm; Waters Technologies, USA) were used to elute the sample at a low flow rate of 0.35 mL/min, which could achieve the rapid separation of S. miltiorrhiza extracts. The elution was joined with DPPH solution (0.3 mL/min) and reacted in the PEEK coil, and then flew into the detector. The anti-oxidation components in S. miltiorrhiza extracts were quickly screened by comparing the chromatograms at 517 nm and 254 nm. Results: The UPLC technology was adopted to realize the efficient separation of samples at low volume flow rate, which avoided the baseline noise caused by high volume flow rate, significantly improved the resolution and sensitivity of the online screening method, and reduced the solvent consumption.The results showed that 31 antioxidant active compounds were screened from S. miltiorrhiza extracts, and 11 of them were identified by PAD-ESI-TOF/MS. Activity verification experiments showed that caffeic acid, salvianolic acid B, salvianolic acid A and danshensu had better antioxidant activity. The IC50 value ofcaffeic acid was 29.00 μmol/L, which was significantly better than the positive control (vitamin C). Conclusion: The method is rapid and efficient, which provides methods and technical support for the rapid discovery of antioxidant components from complex natural products.

Effect of referral on intravascular treatment of acute ischemic stroke with large vessel occlusion / 第二军医大学学报

Objective To explore the impact of referral on intravascular treatment of acute ischemic stroke patients with large vessel occlusion (AIS-LVO), and to analyze the influence factors of prognosis. Methods We retrospectively analyzed the clinical data of the AIS-LVO patients who received intravascular treatment from Sep. 2013 to Feb. 2018 in Stroke Center of our hospital. The patients were divided into directly admitted group and referral group. The patients in the directly admitted group went directly to the Emergency of our hospital through the pre-hospital emergency medical service or other vehicles. The patients in the referral group were transferred from other hospitals to the Emergency of our hospital. The clinical features, curative effect and prognosis were analyzed between the two groups. Then the patients were divided into good prognosis group (modified Rankin scale score at 90 d after operation being 0-2) and poor prognosis group (2). The clinical data and visit methods were analyzed between the good prognosis and poor prognosis groups, and logistic regression analysis was used to analyze the P0.1 variables. Results A total of 316 patients were included, and the directly admitted group had 195 cases (61.7%) and the referral group had 121 cases (38.3%). Compared with the directly admitted group, the proportions of the patients with ischemic stroke and bridging therapy were significantly lower in the referral group (χ2＝4.549, P＝0.033; χ2＝29.319, P0.001). The onset-to-door time (ODT) and onset-to-recanalization time (ORT) were significantly longer in the referral group than those in the directly admitted group (239 [168, 238] min vs 85 [55, 170] min, Z＝1.779, P0.001; 397 [306, 472] min vs 285 [214, 364] min, Z＝6.779, P0.001). The short-term treatment efficiency and good prognosis rate were significantly worse in the referral group than those in the directly admitted group (52.9% [64/121] vs 64.1% [125/195], χ2＝3.903, P＝0.048; 46.3% [56/121] vs 57.9% [113/195], χ2＝4.806, P＝0.043). There were 169 cases (53.5%) in the good prognosis group and 147 cases (46.5%) in the poor prognosis group. Compared with the poor prognosis group, the patients were significantly younger, the proportion of the patients with hyperlipidemia was significantly lower and the proportion of the patients with bridging therapy was significantly higher in the good prognosis group ([64.2±12.8] years vs [69.9±11.9] years, t＝4.095, P0.001; 0.6% [1/169] vs 6.1% [9/147], χ2＝7.848, P＝0.005; 70.4% [119/169] vs 13.6% [20/147], χ2＝102.975, P0.001). Compared with the poor prognosis group, the proportion of directly admitted patients was significantly higher in the good prognosis (66.9% [113/169] vs 55.8% [82/147], χ2＝4.086, P＝0.043), and ODT and ORT were significantly shorter in the good prognosis group (106 [59, 214] min vs 184 [91, 281] min, Z＝3.997, P0.001; 308 [226, 389] min vs 350 [267, 453] min, Z＝2.999, P＝0.003). Logistic regression analysis showed that bridging therapy, direct visit and short ODT were independent predictors of good prognosis in AIS-LVO patients with intravascular treatment. Conclusion The prognosis of referral AIS-LVO patients with intravascular treatment is poorer compared with the directly admitted patients. Bridging therapy, direct visit and short ODT indicate good prognosis in AIS-LVO patients.

Influence of time management on intravenous thrombolysis outcome in acute ischemic stroke / 第二军医大学学报

Objective To analyze the efficacy and prognosis of intravenous thrombolysis in patients with acute ischemic stroke. so as to discuss the influence of time management on the thrombolysis outcome. Methods We retrospectively analyzed the clinical data of the 198 acute ischemic stroke patients who received intravenous thrombolysis in Changhti Hospital from Sep. 2013 to Dec. 2015. The patients were divided into two groups according to U. S. National Institutes of Health Stroke Scale (NIHSS) scores 24 h after thrombolysis: effective group and ineffective group. The clinical data of the two groups were compared to indentify the factors influencing the prognosis. The patients were also divided into two groups according to modified Rankin Scale (mRS) after 90d: good prognosis and poor prognosis groups; the clinical data of the two groups were also analyzed to identify the influencing factors. The differences were compared between the effective group and ineffective group from the onset to different medication periods. so as to evaluate the influence of time management on prognosis. Results The 198 consecutive acute ischemic stroke patients included 74 in the effective group and 124 in the ineffective group. Single-factor analysis indicated that the pre-thrombolysis baseline NIHSS score. time from onset to the beginning of thrombolysis. and history of coronary heart disease were significantly different between the effective group and ineffective group (P<0. 01. P<0. 05). Logistic regression analysis showed that low baseline NIHSS score before thrombolysis. shorter time from onset to the beginning of thrombolysis, and no history of coronary heart disease were the independent predictors of a good effect. Single-factor analysis of indicated that the age, blood glucose level prior to thrombolysis, history of diabetes mellitus, history of a trial fibrillation, baseline NIHSS score and GCS score prior to thrombolysis, time from onset to the beginning of thrombolysis and history of coronary heart disease were significantly different between the good prognosis group (157 cases) and the poor prognosis group (41 cases) (P<0. 01, P<0. 05). Logistic regression analysis showed that low baseline NIHSS score before thrombolysis were independent predictors of good prognosis. Concerning the time management, the time periods from onset to the beginning of thrombolysis were significantly different between the ineffective group and effective group (P<0. 01), which was mainly caused by the time from the onset to treatment (P<0. 01). Conclusion Lower pre-thrombolysis NIHSS score, shorter time from stroke onset to beginning of thrombolysis, no history of coronary heart disease are associated with good short term efficacy, and lower pre-thrombolysis NIHSS score is the independent factor of good prognosis for acute ischemic stroke. Early treatment may increase the efficacy of intravenous thrombolysis.