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Medical Writing:
Published in Lynne M. Bianchi, Research during Medical Residency, 2022
Lynne M. Bianchi, Randy Jeffrey
For common methods, you might cite another paper that details the technique, but first confirm that the methods were the same as those you used. If necessary, describe the modifications you made to the existing protocol. For example, “Biopsy samples were processed as in Liu et al (2002), except that tissues were fixed in 10 percent formalin rather than 4 percent paraformaldehyde.”
RGS7 silence protects palmitic acid-induced pancreatic β-cell injury by inactivating the chemokine signaling pathway
Published in Autoimmunity, 2023
Yurong Zhu, Jun Li, Tao Ba, Yuan Sun, Xiangyun Chang
The cell proliferation was analyzed by using the 5-ethynyl-2′- deoxyuridine (EdU) Cell Proliferation Kit (C0078S, Beyotime). Beta-TC-6 and Min6 cells (2 × 104 cells/well) were cultured in 24-well plates overnight. Cells were incubated with EdU solution (20 μM) for 2 h. Subsequently, 4% paraformaldehyde was added for 15 min at room temperature. After incubating with PBS for 10–15 min at room temperature, cells were incubated with endogenous peroxidase blocking solution for 20 min to inactivate endogenous peroxidase. After three times wash with PBS, 4′,6-diamidino-2-phenylindole (DAPI) was added and incubated for 10 min in a dark room at room temperature to stain cell nuclei. Finally, EdU-positive cells were observed and counted with a fluorescence microscope. All the experiments were performed in triplicate.
MiR-5622-3p inhibits ZCWPW1 to induce apoptosis in silica-exposed mice and spermatocyte cells
Published in Nanotoxicology, 2023
Moxuan Zhao, Guiqing Zhou, Jingjing Wang, Yue Zhang, Jinglong Xue, Jianhui Liu, Junhong Xie, Lihua Ren, Xianqing Zhou
4% paraformaldehyde solution was applied to fix testes tissue and cells. For testes, they were embedded into paraffin blocks and cut into sections. After being heated for 2 h at 60 °C, these sections were dewaxed by xylene 3 times for 10 minutes each and hydrated in gradients of ethanol (100%, 95%, 85%, 75%) and in phosphate buffer saline (PBS) thrice for 5 minutes each. Both tissues and cells were treated with 0.05% (v/v) Triton X-100 for 40 min and then blocked with fetal bovine serum (5%). After that, they were covered by γ-H2AX rabbit polyclonal antibody (1:200, CST, USA) at 4 °C for the whole night. These samples were covered with anti-rabbit IgG secondary antibody (1:300, CST, USA) for 1.5h at 25 °C the next day. DAPI was used to stain nuclei. Laser scanning confocal microscopy was applied for the image.
XiaoEr LianHuaQinqGan alleviates viral pneumonia in mice infected by influenza A and respiratory syncytial viruses
Published in Pharmaceutical Biology, 2022
Wenyan Li, Tongtong Li, Chi Zhao, Tao Song, Yao Mi, Zhang Chuangfeng, Yunlong Hou, Zhenhua Jia
The immunofluorescence assay was performed to test the inhibitory effect of XELH on viral nucleoprotein (NP) expression in vitro. MDCK cells were incubated in 8-well plates, washed with PBS, and then exposed to 1 MOI H3N2 for 2 h at 4 °C. The cells were washed with PBS and a medium containing TPCK treated-trypsin (Sigma-Aldrich, Germany; cat. no. 232-650-8). XELH (1 mg/mL) containing TPCK was added 0, 2, 4, 6, and 8 h after infection. After 10 h, the slides were washed twice with PBS. Next, 4% paraformaldehyde was added for 20 min at room temperature (25 °C). The sections were then washed three times with PBS, incubated with 0.5% Triton-X100 for 10 min at room temperature, and washed three times with PBS. Next, 5% goat serum was added for 20 min at room temperature. The sections were incubated with anti-influenza A virus NP antibody (Abcam; cat. no. ab20343) at 4 °C overnight, followed by incubation with a fluorescein-conjugated affinipure goat anti-mouse IgG antibody (Proteintech; cat. no. SA00003-1) for 1 h at room temperature. Finally, the cells were labelled with 4′,6-diamidino-2-phenylindole (DAPI) glycerin-staining solution and observed under a fluorescence microscope (Nikon 942101).