Cu(OAc)2-Promoted Oxidative Cross-Dehydrogenative Coupling Reaction of α-Acylmethyl Malonates with Indole Derivatives to Access 3-Functionalized Indoles and Polycyclic Indoles.

A Cu(OAc)2-promoted oxidative cross-dehydrogenative coupling reaction of α-acylmethyl malonates with indole derivatives was developed. In the case of indoles, the regioselective coupling products were formed through a sequential dehydrogenation-addition-dehydrogenation process. When a second nucleophilic center was located in the 2-position of indoles, further successive nucleophilic cyclization occurred to give polycyclic indole derivatives. The Cu(OAc)2 was proved to act as not only an oxidant but also a catalyst.

Intracellular Zinc Quantification by Fluorescence Imaging with a FRET System.

Fluorescence imaging of cellular metals is widely reported. However, the quantification of intracellular metals with fluorescence imaging is so far not feasible and highly challenging. In this work, a ratiometric probe with two fluorescently labeled complementary DNA strains is designed for intracellular zinc quantification via fluorescence imaging, based on fluorescence resonance energy transfer (FRET) from carbon dots (CDs) to fluorescein (FAM). The donor CDs are modified with a Zn2+ aptamer, whereas the receptor FAM is conjugated with the complementary DNA sequence to ensure selectivity. MCF-7 cells are cultured sequentially with Zn2+ (20, 40, 55, 70, 85, and 100 µmol L-1) and CDs-FAM (100 µg mL-1), which is used for fluorescence imaging (at λex = 405 nm and λem = 440-490 nm for CDs, λem = 500-550 nm for FAM) to provide a relative fluorescence ratio (( F - F0)/ F0, F = ICDs/ IFAM), followed by quantifying intracellular zinc with ICPMS. A linear correlation is achieved between the relative fluorescence ratio in fluorescence images and the intracellular zinc content derived by ICPMS, which facilitates intracellular zinc quantification via fluorescence imaging. It is especially useful for real-time tracing of intracellular zinc during the cell culturing process or in vivo. The cellular uptake of Zn2+ by MCF-7 cells is further evaluated with this approach by culturing with 100 µmol L-1 of Zn2+ for different times, and a maximum uptake of 60.5 fg per cell is observed at an incubation time of 60 min. This value is further demonstrated well by ICPMS detection.

Sacral injury and influencing factors after ultrasonic ablation of uterine fibroids ≤30 mm from the sacrum

PURPOSE: To study sacral injuries and influencing factors after ultrasonic ablation of uterine fibroids no more than 30 mm from the sacrum. METHODS: A total of 406 patients with uterine fibroids who underwent percutaneous ultrasound ablation were analyzed retrospectively. All patients underwent contrast-enhanced magnetic resonance imaging (MRI) scans before and after high-intensity focused ultrasound. The abnormal signal intensity (low signal intensity on T1WI and high signal intensity on T2WI) on the postoperative MRIs was indicative of a sacral injury. The patients were divided into a sacrum injury group and a sacrum non-injury group. The relationship between fibroid characteristics, ultrasound ablation parameters, and injury was analyzed using univariate and multivariate analyses. RESULTS: There were 139 cases of sacral injury (34.24%). When the distance from the fibroid's dorsal side to the sacrum was 0-10 mm, the risk assessment showed that the danger of sacral injury increased by 1.85 times and 3.03 times compared with that at a distance of 11-20 or 21-30 mm. Furthermore, the risk of sacral injury increased by 1.89 times and 3.23 times when the therapeutic dose (TD) of a fibroid was >500 KJ compared with that of a fibroid with TD= 250-500 KJ and <250 KJ. CONCLUSION: A distance of 10 mm or less and a TD of >500 KJ were significantly correlated with sacral injury. The distance from the fibroid's dorsal side to the sacrum and the TD were the main causes of injury to the sacrum. A distance of 10 mm or less and a TD of >500 KJ carried higher injury risks, while a distance of 21-30 mm and a TD of <250 KJ were the most appropriate circumstances to reduce the risk of sacral injury.

Upregulation of FAM50A promotes cancer development.

FAM50A encodes a nuclear protein involved in mRNA processing; however, its role in cancer development remains unclear. Herein, we conducted an integrative pan-cancer analysis using The Cancer Genome Atlas, Genotype-Tissue Expression, and the Clinical Proteomic Tumor Analysis Consortium databases. Based on the gene expression data from TCGA and GTEx databases, we compared FAM50A mRNA levels in 33 types of human cancer tissues to those in corresponding normal tissues and found that FAM50A mRNA level was upregulated in 20 of the 33 types of common cancer tissues. Then, we compared the DNA methylation status of the FAM50A promoter in tumor tissues to that in corresponding normal tissues. FAM50A upregulation was accompanied by promoter hypomethylation in 8 of the 20 types of tumor tissues, suggesting that promoter hypomethylation contributes to the upregulation of FAM50A in these cancer tissues. Elevated FAM50A expression in 10 types of cancer tissues was associated with poor prognosis in patients with cancer. FAM50A expression was positively correlated with CD4+ T-lymphocyte and dendritic cell infiltration in cancer tissues but was negatively correlated with CD8+ T-cell infiltration in cancer tissues. FAM50A knockdown caused DNA damage, induced interferon beta and interleukin-6 expression, and repressed the proliferation, invasion, and migration of cancer cells. Our findings indicate that FAM50A might be useful in cancer detection, reveal insights into its role in cancer development, and may contribute to the development of cancer diagnostics and treatments.

A fluorescence imaging protocol for correlating intracellular free cationic copper to the total uptaken copper by live cells.

A variety of fluorescence probes have been developed for fluorescence imaging of metals in biological cells. However, accurate quantification of metals with fluorescent approaches is challenging due to the difficulty in establishing a standard calibration curve in living cells. Herein, a fluorescence imaging protocol is developed for imaging intracellular Cu2+ and its correlation with the cellular uptake of copper. The total amount of intracellular Cu is detected by inductively coupled plasma mass spectrometry (ICP-MS) in parallel. Fluorescence imaging of Cu2+ is accomplished with Rhodamine B derivative modified carbon dots (CDs-Rbh) based on fluorescence resonance energy transfer (FRET) from CDs to rhodamine. Intracellular Cu2+ is correlated with fluorescence ratio at λem 500-600 nm (rhodamine) to λem 425-475 nm (CDs) with excitation at λex 405 nm. It is found that Cu2+ is linearly correlated with the total intracellular uptaken copper content, with a linear correlation between the relative fluorescence ratio in fluorescence imaging and intracellular Cu derived from ICP-MS, including both Cu(I) and Cu(II) species. The linear calibration equation is lg(F2/F1) = 0.00148 m[Cu]-0.3622. This approach facilitates further investigation and elucidation of copper transition in live cells and the evaluation of their cytotoxicity.

Simultaneous detection and speciation of mono- and di-valent copper ions with a dual-channel fluorescent nanoprobe.

Redox cyclings between mono-/di-valent copper oxidation states occur in electron transfer reactions that underlie their biological functions. We report herein a dual channel fluorescent nanoprobe for the detection of mono- and di-valent copper ions. The probe, BSA-CDs@RBH/BCS, is designed by decorating carbon dots (CDs) on BSA encapsulated rhodamine hydrazide (RBH) and conjugating with bathocuproine disulfonate (BCS). Cu2+ catalyzes the hydrolysis of RBH, and the formed rhodamine B (RhB) shows emission at λex/λem = 360/575 nm which ensures Cu2+ detection. BSA reduces Cu2+ to Cu+ and the BCS-Cu+ chelate shows emission at λex/λem 360/450 nm which ensures Cu+ assay. Thus, the dual-channel fluorescence enables speciation of Cu2+ and Cu+.

Copper-Catalyzed Annulation of Oxime Acetates with α-Amino Acid Ester Derivatives: Synthesis of 3-Sulfonamido/Imino 4-Pyrrolin-2-ones.

A copper-catalyzed annulation of oxime acetates and α-amino acid ester derivatives for the easy preparation of 4-pyrrolin-2-ones bearing a 3-amino group has been developed. This process features the oxidation of amines with oxime esters as the internal oxidant to produce the active 1,3-dinucleophilic and 1,2-dielectrophilic species concurrently. The subsequent nucleophilic cyclization realizes the efficient construction of 4-pyrrolin-2-one derivatives.

Glutathione triggered degradation of polydopamine to facilitate controlled drug release for synergic combinational cancer treatment.

Here we report a novel mechanism for triggering drug release in the polydopamine (PDA)-coated magnetic CuCo2S4 core-shell nanostructure by glutathione (GSH) triggered degradation of PDA for release. In the design, we used PDA coated CuCo2S4 as the nanocarrier with polyethylene glycol and folic acid targeting molecules to ensure the safe delivery of doxorubicin (DOX) to cancer cells. In addition, the controlled release could be enforced by taking advantage of the pH sensitivity of PDA to tumor acidic environments. The targeting and treatment of HeLa cancer cells were very effective and the killing was more efficient at higher levels of GSH. Furthermore, the designed system not only could be used for drug delivery but also could combine photothermal therapy with chemotherapy in a synergetic way. Plus, the system could be used for magnetic resonance imaging (MRI), which is beneficial for imaging-guided treatment.

Cu(OAc)2-Triggered Cascade Reaction of Malonate-Tethered Acyl Oximes with Indoles, Indole-2-alcohols, and Indole-2-carboxamides.

A Cu(OAc)2-promoted cascade reaction of malonate-tetherd acyl oximes with indoles, indole-2-alcohols, or indole-2-carboxmides provides facile access to polysubstituted 3-pyrrolin-2-ones. The reaction features the generation of two adjacent electrophilic centers at the same time as cyclization to lactam. The subsequent double addition with nucleophiles followed by oxidation realizes the difunctionalization of the imine sp2-carbon and the adjacent α-sp3-carbon.

Probing pH variation in living cells and assaying hemoglobin in blood with nitrogen enriched carbon dots.

Nitrogen enriched carbon dots (CDs) are prepared by hydrothermal treatment with oxalic acid and ethylenediamine serving as raw materials. The obtained CDs display strong blue fluorescence emission at λex/λem = 320/410 nm, along with a quantum yield of 14%. It is interesting to obtain a linear increase of fluorescence with the variation of pH value within pH 5.01-8.94. Meanwhile, a favorable stability of the fluorescence is achieved in a high saline matrix, i.e., up to 1.0 mol L-1 NaCl, making the CDs particularly suitable for probing pH variation in living cells or real biological systems. In addition, hemoglobin (Hb) significantly quenched the fluorescence of CDs due to the inner filter effect and static quenching, providing a linear relationship between fluorescence and Hb concentration within 20-500 mg L-1. The CDs exhibit favorable selectivity for both pH sensing and hemoglobin detection, which are further demonstrated by selective and sensitive intracellular pH monitoring in living HeLa cells and Hb assay in human blood.

Deep Eutectic Solvent-Assisted Preparation of Nitrogen/Chloride-Doped Carbon Dots for Intracellular Biological Sensing and Live Cell Imaging.

A novel approach for the preparation of dual-functional carbon dots, i.e., nitrogen- and chloride-doped carbon dots, abbreviated as N/Cl-CDs, is developed with the assistance of a choline chloride-glycerine deep eutectic solvent (DES). The carbon source is provided by urea and the DES serves as a solvent for controlling the preparation of CDs in the absence of water. The dual-element doped carbon dots are oxygen-rich with hydroxyl and amine groups. They exhibit an average particle size of ca. 3.88 nm and give rise to strong and pH-sensitive fluorescent emission at λex/λem = 340/430 nm with a quantum yield of 16.15 ± 1.36%. It is particularly interesting to see that the fluorescence of N/Cl-CDs remains stable in a high-salinity matrix, providing vast potentials for treating real biological sample matrixes with high salinity. The N/Cl-CDs provide an optical probe for intracellular pH sensing and multicolor imaging in HeLa cells. In addition, the N/Cl-CDs show obvious fluorescence response to cytochrome c (cyt- c) with a detection limit of 3.6 mg L-1 (ca. 0.29 µmol L-1) within in a range of 10-500 mg L-1, providing potentials for fluorescence detection of cyt- c as well as facilitating intracellular cyt- c imaging.

Dual Functional Core-Shell Fluorescent Ag2S@Carbon Nanostructure for Selective Assay of E. coli O157:H7 and Bactericidal Treatment.

A dual functional fluorescent core-shell Ag2S@Carbon nanostructure is prepared by a hydrothermally assisted multi-amino synthesis approach with folic acid (FA), polyethylenimine (PEI), and mannoses (Mans) as carbon and nitrogen sources (FA-PEI-Mans-Ag2S nanocomposite shortly as Ag2S@C). The nanostructure exhibits strong fluorescent emission at λex/λem = 340/450 nm with a quantum yield of 12.57 ± 0.52%. Ag2S@C is bound to E. coli O157:H7 via strong interaction with the Mans moiety in Ag2S@C with FimH proteins on the fimbriae tip in E. coli O157:H7. Fluorescence emission from Ag2S@C/E. coli conjugate is closely related to the content of E. coli O157:H7. Thus, a novel procedure for fluorescence assay of E. coli O157:H7 is developed, offering a detection limit of 330 cfu mL-1. Meanwhile, the Ag2S@C nanostructure exhibits excellent antibacterial performance against E. coli O157:H7. A 99.9% sterilization rate can be readily achieved for E. coli O157:H7 at a concentration of 106-107 cfu mL-1 with 3.3 or 10 µg mL-1 of Ag2S@C with an interaction time of 5 or 0.5 min, respectively.