All the mice were sacrificed on day 38, and they were weighed and then euthanized by exsanguination after the intravenous administration of 10% chloral hydrate solution (100 mg chloral hydrate/kg body weight; Sigma, USA) and immediately dissected. Blood samples were collected by eyeball enucleation, and the plasma samples were prepared and stored at −80 °C until the analyses. The liver, thymus, kidneys, and spleen were collected and weighed immediately. The organ indexes were expressed relative to the body weight (kg of organ/kg of body weight × 100). Each intestine segment was divided into two parts, one part was firstly used to collect intestinal contents instantly, and the other part was then used to collect intestinal tissue. Here, the contents of the duodenum, jejunum, and ileum were immediately collected and mixed as the small intestine contents, and the contents of the cecum and colon were immediately collected and stored directly. All contents were stored in liquid nitrogen for 24 h, and then transfer to -80 °C until further DNA extraction and intestinal fermentation detection. Then, the tissues (about 1 cm in length) from the duodenum, jejunum, ileum, and colon tissue whose contents were removed and washed with PBS buffer, were immediately stored in liquid nitrogen for 24 h, and then transfer to -80 °C until further RNA extraction.
Chloral hydrate solution
Manufactured by Merck Group
Sourced in Canada
Chloral hydrate solution is a laboratory chemical used as a reagent. It is a clear, colorless liquid with a distinctive odor. The solution is commonly utilized in various chemical and research applications, where it serves as a precursor or intermediate in the synthesis of other compounds.
Automatically generated - may contain errors
Lab products found in correlation
Hydrochloric acid (hcl), by Thermo Fisher Scientific (3 mentions)
Formic acid, by Thermo Fisher Scientific (3 mentions)
Aerrane, by Baxter (3 mentions)
Xylazine, by Bayer (3 mentions)
Aluminium chloride hexahydrate, by Merck Group (2 mentions)
Chloral hydrate, by Merck Group (1 mentions)
Toluidine blue, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Aluminum chloride hexahydrate, by Merck Group (1 mentions)
6 protocols using chloral hydrate solution
1
Tissue and Content Sampling Protocol
2
Histological Analysis of Guinea Pig Nasal and Lung Tissues
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The guinea pigs were deeply anesthetized with 10% chloral hydrate solution (300 mg/kg; Sigma-Aldrich; Merck KGaA) by intraperitoneal injection. The nasal bone of each guinea pig was removed and the nasal mucosa was gained. The lung was removed. Nasal mucosa and lung tissues were fixed in 4% paraformaldehyde at 4°C for 48 h. Nasal mucosa was decalcified in 10% EDTA (Sigma-Aldrich; Merck KGaA). The tissues were embedded in paraffin and sliced at a thickness of 4–5 µm. The sections were stained with hematoxylin and eosin (H&E) or toluidine blue (Sigma-Aldrich; Merck KGaA). Collagen deposition in lung tissues was observed by Masson's trichrome stain. Five microscopic fields were randomly selected, and the eosinophils and mast cells in each high-power microscopic field (magnification, ×400) were counted under a light microscope.
3
Histological Analysis of Decalcified Bone
4
Histological Analysis of Decalcified Bone
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Animals were anesthetized with a mixture of a 20% chloral hydrate solution (0.4 mg/g body weight Sigma-Aldrich Canada Ltd, Oakville, ON, Canada) and Rompun (0.005 mg/g body weight; xylazine; Bayer Inc) and sacrificed by an inhalation overdose of AErrane (isoflurane USP, Baxter). Tibias were dissected and immersed in a fixative solution consisting of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.2 overnight at 4 °C. The samples were then decalcified for 3 days at 4 °C in Planck-Rychlo solution consisting of 0.13 M aluminium chloride hexahydrate (Sigma-Aldrich), 0.2 N hydrochloric acid (Fisher Scientific, Whitby, ON, Canada), 1.35% formic acid (Fisher Scientific) and then the implants were removed carefully. Decalcified samples were washed for 24 h in 0.1 M phosphate buffer (pH 7.2), dehydrated through graded ethanols, cleared with xylene, embedded in paraffin and serially-sectioned at 5 μm thickness. Some sections were stained with hematoxylin and eosin and others with toluidine blue for observation by light microscopy and histomorphometric analyses.
5
Histological Analysis of Maxillary Samples
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Animals were anesthetized with a mixture of a 20% chloral hydrate solution (0.4 mg/g body weight Sigma‐Aldrich Canada Ltd, Oakville, ON, Canada) and Rompun (0.005 mg/g body weight; xylazine; Bayer Inc.), and sacrificed by an inhalation overdose of AErrane (isoflurane USP, Baxter). Maxillae were dissected and immersed overnight at 4°C in a fixative solution consisting of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.2. The samples were then decalcified for 3 days at 4°C in Planck‐Rychlo solution consisting of 0.13 M aluminum chloride hexahydrate (Sigma‐Aldrich), 0.2 N hydrochloric acid (Fisher Scientific, Whitby, ON, Canada), 1.35% formic acid (Fisher Scientific). Decalcified samples were washed for 24 hr in 0.1 M phosphate buffer (pH 7.2), dehydrated through graded ethanols, cleared with xylene, embedded in paraffin and sectioned at 5 μm thickness. All the sections were cut in a mesial‐distal orientation. The sections were stained with hematoxylin and eosin for observation by light microscopy.
6
Prostate Cancer Model in Mice
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C57BL/6 black mice (8 weeks old; male n=25; female n=25; weight ~20–25 g), were purchased from Sun Yat-sen University Experimental Animal Center (Guangzhou, China) and were used as control mice. A total of four B6.Cg-Selplgtm1Fur/J mice, (8 weeks old; male n=2; female n=2; weight ~20–25 g), which highly express CXCL9, were purchased from Jackson Laboratory (Ben Harbor, ME, USA). In total, 54 mice were used in this study. They were maintained under specific pathogen-free conditions with 12-h light/12-h dark cycles at 26–28°C and 50–65% humidity and a normal diet (including 4% fat and 0.07% cholesterol) and water ad libitum.
DMAB (3,2′-dimethyl 4-aminobiphenyl), was used as a chemical carcinogenic agent. The mice were enrolled and initiation with subcutaneous injection of DMAB (150 mg/kg b.w.) once a week for 3 weeks to construct the prostate cancer model. The mice were anesthetized by 10% chloral hydrate solution (3 ml/kg, Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), and then the blood samples were collected. Then the mice were sacrificed by cervical dislocation, and the prostate tissues were collected and stored. Animal experiments were approved by the Medical Ethics Committee of Linyi People's Hospital.
DMAB (3,2′-dimethyl 4-aminobiphenyl), was used as a chemical carcinogenic agent. The mice were enrolled and initiation with subcutaneous injection of DMAB (150 mg/kg b.w.) once a week for 3 weeks to construct the prostate cancer model. The mice were anesthetized by 10% chloral hydrate solution (3 ml/kg, Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), and then the blood samples were collected. Then the mice were sacrificed by cervical dislocation, and the prostate tissues were collected and stored. Animal experiments were approved by the Medical Ethics Committee of Linyi People's Hospital.
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