Pathological features and immune microenvironment in HER-2 intratumoral heterogeneous breast cancers / 中华肿瘤杂志

Objective: To observe the clinical pathology features, and immune microenvironment of HER-2 intratumoral heterogeneity breast cancer. Methods: Thirty cases of HER-2 intratumoral heterogeneous breast cancer were retrospectively analyzed in Tianjin Medical University Cancer Institute and Hospital from November 2017 to June 2020. HER-2 expression was detected by immunohistochemistry and verified by dual color silver-enhanced in-situ hybridization (D-SISH). HER-2 intratumoral positive and negative regions were divided. The pathological characteristics, subtype, and the level of tumor infiltrating lymphocytes (TILs) and the expression of programmed cell death-ligand 1 (PD-L1) were evaluated respectively. Results: The proportion of HER-2 positive cells of the breast cancer ranged from 10% to 90%. The pathological type was mainly invasive non-special typecarcinoma. Six cases presented different pathological types between HER-2 positive and negative regions. The HER-2-positive areas included 2 cases of carcinoma with apocrine differentiation, and the negative areas included 2 cases of invasive micropapillary carcinoma, 1 case of invasive papillary carcinoma, and 1 case of carcinoma with apocrine differentiation. In HER-2 positive regions, 17 cases were Luminal B and 13 cases were HER-2 overexpressed types. There were 22 cases of Luminal B and 8 cases of triple negative tumors in the HER-2 negative areas. The levels of TILs in HER-2 positive and negative areas accounted for 53.3% (16/30) and 26.7% (8/30), respectively, with a statistically significant difference (P=0.035). The positive expression of PD-L1 in HER-2 positive area and HER-2 negative area were 6 cases and 9 cases, respectively. Among 8 cases with HER-2 negative regions containing triple negative components, 4 cases were positive for PD-L1 expression. Conclusions: In the case of HER-2 intratumoral heterogeneity, it is necessary to pay attention to both HER-2 positive and negative regions, and evaluate subtype separately as far as possible. For HER-2 intratumoral heterogeneous breast cancer containing triple negative components, the treatment mode can be optimized by refining the intratumoral expression of PD-L1.

Detection of Synthetic Cannabinoid CUMYL-PEGACLONE in E-Cigarette Oil and Hair / 法医学杂志

OBJECTIVES@#To establish a combined high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) method to detect the synthetic cannabinoid CUMYL-PEGACLONE in e-cigarette oil and hair.@*METHODS@#HPLC-MS/MS and GC-MS were used to establish the detection method of CUMYL-PEGACLONE, and the hair of drug-involved persons and the seized e-cigarette oil were detected.@*RESULTS@#The main mass spectrometry characteristic ions m/z of CUMYL-PEGACLONE measured by GC-MS were 91, 179, 197, 254 and 372. CUMYL-PEGACLONE had a good linear relationship in the mass concentration range of 2-50 ng/mL, and the linear correlation coefficient (r) was greater than 0.99. The limit of detection (LOD) of CUMYL-PEGACLONE in hair was 0.01 ng/mg, and the limit of quantitation (LOQ) was 0.02 ng/mg. The LOD of CUMYL-PEGACLONE in e-cigarette oil was 1 ng/mg, and the LOQ was 2 ng/mg. The average recoveries of CUMYL-PEGACLONE under the attempt at high, intermediate and low levels in blank human hair and e-cigarette oil matrix were 98.2%-132.4% and 93.5%-110.6%, and the intraday and intraday precision were 1.2%-12.9% and 0.7%-2.9%. CUMYL-PEGACLONE was detected in the hair of 15 drug-involved persons. Except for 1 person who was lower than LOQ, the concentration of CUMYL-PEGACLONE in the hair of other 14 persons was 0.035-0.563 ng/mg. The mass fraction of CUMYL-PEGACLONE in 2 e-cigarette oil were 0.17% and 0.21%, respectively.@*CONCLUSIONS@#The established HPLC-MS/MS and GC-MS methods are applied to the detection of HPLC-MS/MS in drug-related cases, which provides strong evidence support for the handling authority to quickly investigate these cases, and also provides a reference for the identification of such substances in future.

SDF-1α/CXCR4 axis promotes migration and invasion of pancreatic cancer cells through inducing epithelial-mesenchymal transition / 中国病理生理杂志

AIM:To investigate the role of SDF-1α/CXCR4 axis in pancreatic cancer cell migration and invasion.METHODS:The mRNA expression of CXCR4 in 4 pancreatic cancer cell lines was detected by RT-qPCR.The migration and invasion abilities of PANC-1 cells with the axis activated by exogenous SDF-1αor inhibited by CXCR4 inhibitor AMD3100 were detected by Transwell assays.The cell viability was measured by MTS assay.The protein expression of the epithelial-mesenchymal transition (EMT) -related molecules in the cells treated with exogenous SDF-1αor AMD3100 was determined by Western blot.RESULTS:All of the 4 pancreatic cancer cell lines expressed CXCR4 mRNA, while the PANC-1 cell line expressed the most.Exogenous SDF-1αpromoted the migration and invasion abilities of PANC-1 cells, which was inhibited by AMD3100.The PANC-1 cells treated with exogenous SDF-1αfor 72 h grew faster, while SDF-1αcombined with AMD3100 made little significance to the viability of PANC-1 cells.Exogenous SDF-1αinduced EMT of PANC-1 cells by up-regulating the expression of SNAIL and TWIST, and AMD3100 reversed this effect.CONCLUSION:SDF-1α/CXCR4 axis enhances the migration and invasion abilities of pancreatic cancer cells through inducing EMT.

Effects of Aconitum coreanum polysaccharides on TGF-β induced endothelial-mesenchymal transition and its mechanisms / 中国药理学通报

Aim To investigate the protective effect of Aconitum coreanum polysaccharides ( ACP) on TGF-β induced endothelial-mesenchymal transition ( EndMT) and its underlying mechanism. Methods HUVECs EndMT model was reproduced by 10 μg·L-1 TGF-β cultured for 72 h in vitro. The effect of ACP(50, 100, 200 mg·L-1) at various concentrations on TGF-β in-duced EndMT of HUVECs was explored. CCK8 assay was employed to detect the cell viability of HUVECs stimulated by ACP. Scratch test was used to analyze the migration ability of cells. The expressions of CD31, α-SMA, Smad2/3, p-Smad2/3 were deter-mined by Western blot. Results CCK-8 assay showed that various concentrations of ACP could promote HU-VECs viability obviously by 10 μg·L-1 TGF-β cul- tured for 36 h. Scratch test showed that TGF-β en-hanced transfer ability of HUVECs significantly, while this case was apparently reversed under the costimula-tion of TGF-β and ACP. Western blot showed that CD31 expression decreased significantly, and the ex-pressions of α-SMA, p-Smad2, p-Smad3 increased significantly, and this phenomenon could be regressed with ACP. Conclusion ACP can inhibit EndMT in HUVECs induced by TGF-β clearly, and the mecha-nism is related to down-regulation of p-Smad2, p-Smad3.

Role of calcium-activated chloride channels in the regulation of pulmonary vascular tone in rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the role of calcium-activated chloride channels and the Cl- channel blockers niflumic acid (NFA) and indanyloxyacetic acid (IAA-94) in the regulation of vascular contraction induced by phenylephrine (PE).</p><p><b>METHODS</b>The PE-induced contraction in rat pulmonary artery was observed by using routine blood vascular perfusion in vitro. The fluorescence Ca2+ indicator Fura-2/AM was used to observe intracellular free Ca2+ concentration ([Ca2+]i) of rat pulmonary artery smooth muscle cells (PASMCs) which were obtained by the acute enzyme separation method (collagenase I plus papain) on NFA and IAA-94 effects on PE-induced contraction. Changes of [Ca2+]i in PASMCs were measured by spectrofluorometry.</p><p><b>RESULTS</b>The anion channel blockers NFA and IAA-94 produced inhibitory effects on PE-induced contractions in the pulmonary artery. NFA and IAA-94 negligibly affected the KCl-induced pulmonary artery contractions. PE could increase [Ca2+]i but NFA and IAA-94 negligibly affected it.</p><p><b>CONCLUSION</b>Calcium-activated chloride channels contribute to the agonist-induced pulmonary artery contractions under physiological conditions, which may be a new clue to investigate the hypoxic pulmonary vasoconstriction.</p>