The following reagents were used: CBD (Sigma, United States); colchicine (Beyotime, China); CCl4 (Aladdin, China); peanut oil (Yuanye, China); ELISA kits (Novus, United States); hematoxylin and eosin (HE) and Masson assay kits (Beyotime, China); aspartate aminotransferase (AST) and hyaluronic acid (HA) kits (Nanjing Jiancheng Bioengineering Institute, China); RIPA lysis buffer and a BCA kit (Solaibao, China); COX-2, p-IκBα, and IκBα antibodies (Wanleibio, China); p-NF-κB, NF-κB, p-p38 MAPK, and p-38 MAPK antibodies (CST, United States); TGF-β1, α-SMA, COL-I, PPAR-α and GAPDH antibodies (Abcam, United States); horseradish peroxidase-labeled secondary antibodies (Bioprimacy, China); chemiluminescence (ECL) color developing solution (Merck, United States); an RNeasy mini kit (Axygen, United States); a PrimeScript RT reagent kit with gDNA Eraser and SYBR Green Master Mix (Takara, Japan); an automatic chemical analyzer (Hitachi, Japan); and light microscopy (Nikon, Japan).
Automatic chemical analyzer
Manufactured by Hitachi
Sourced in Japan
The Automatic Chemical Analyzer is a laboratory instrument designed to perform automated analysis of chemical compounds. It is capable of processing multiple samples and conducting various tests, including colorimetric, enzymatic, and ion-selective analyses. The core function of the Automatic Chemical Analyzer is to provide consistent, reliable, and efficient sample analysis in a laboratory setting.
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4 protocols using automatic chemical analyzer
1
Hepatoprotective Effects of CBD
2
Minimally Invasive Surgical Outcomes
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The length of the skin incision, operation time, intraoperative blood loss, and postoperative blood transfusion volume were evaluated between the groups. A visual analog pain scale was recorded. Serological markers indicating muscle injury, including creatinine kinase (CK) and aldolase, the pro-inflammatory cytokines interleukin (IL)-6 and IL-8, and the anti-inflammatory cytokines IL-10 and IL-1 receptor antagonist (ra) were measured to assess the degree of tissue injury and the systemic inflammatory reaction. All evaluations were performed at 7:00 AM prior to the operation and on postoperative days (POD) 1, 3, 7, and 14. Serum CK was analyzed by the ultraviolet kinetic method with an automatic chemical analyzer (Hitachi Inc., Tokyo, Japan), and serum aldolase was analyzed by the ultraviolet kinetic method with a spectrophotometer (Sentinel, Rome, Italy). The cytokines were analyzed with a commercially available enzyme linked immunosorbent assay test kit (Quantikine, R&D Systems, Minneapolis, MN, USA). Results were analyzed with the Mann-Whitney U-test and SPSS ver. 11.5 (SPSS Inc., Chicago, IL, USA). Statistical significance was tested with a 95% confidence interval. A p < 0.05 was considered significant.
3
Colon Tissue Cytokine and Antioxidant Analysis
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Ten milligrams of colon tissue was ground in liquid nitrogen and lysed by using RIPA lysis (CST, Danvers, MA, USA). The supernatant was prepared via centrifugation at 12,000×g for 10 min and stored at −20°C for ELISA measurement. The levels of tumor necrosis factor α (TNFα) (ab100747), interleukin (IL)-1 β (ab100704), IL-6 (ab100713) and IL-10 (ab100697) were measured by using the ELISA kits from Abcam (San Francisco, CA, USA). The levels of superoxide dismutase (SOD) (#MBS080359) and catalase (CAT) (#MBS775862) and glutathione peroxidase (GSPx) (#MBS049725) were also evaluated using the kits from MyBioSource, Inc. (San Diego, CA, USA). All biochemical indexes were measured on an automatic chemical analyzer (Hitachi, Tokyo, Japan). MDA was measured by using UV–Vis spectroscopy at 267 nm in 10 mM Na3PO4 buffer pH 11 according to the molar absorbing coefficient 31,500 M−1cm−1 (Esterbauer et al., 1991 (link)). Wavelength scanning of MDA fractions was conducted via the wavelength against a control group without 1,3-propanediol and MDA.
4
Oxidative Stress and Cytokine Profiling
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Serum levels of malondialdehyde (MDA) (MBS269473), superoxide dismutase (SOD) (#MBS080359), catalase (CAT) (#MBS775862), and glutathione peroxidase (GSPx) (#MBS049725) were also evaluated using the kits from MyBioSource, Inc. (San Diego, CA, USA). The serum levels of tumor necrosis factor α (TNFα) (ab100747), interleukin- (IL-) 1 β (ab100704), IL-6(ab100713), and IL-10 (ab100697) were measured by using the ELISA kits from Abcam (San Francisco, CA, USA). All biochemical indexes were measured on an automatic chemical analyzer (Hitachi, Tokyo, Japan).
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