Application of Grb10 as key negative regulation factor of beta cell dysfunction
A dysfunctional, β-cell technology, applied in the field of biomedicine, can solve problems such as unclear specific mechanisms
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Embodiment 1
[0045] Embodiment 1 human body study
[0046] The study used islets from 13 human organ donors, including 6 minors (under 18 years old) and 7 adults (over 18 years old). Grb10 expression was compared between islets younger than 18 years old and adult donors (greater than 18 years old). Human pancreases were obtained from deceased non-diabetic donors by the Department of Urology, Department of Organ / Liver Transplantation, Second Xiangya Hospital, Central South University.
[0047] Use collagenase P to digest and isolate human islets, carefully clean the adipose tissue, lymph nodes, blood vessels and fascia around the pancreas, trim the human pancreas, inflate the pancreatic duct with a 5ml syringe, and inject 1mg / mL collagenase P through the pancreatic duct Hanks Balanced Salt Solution (HBSS), the amount of collagenase solution used is equivalent to twice the weight of the pancreas. The pancreas was taken out and placed in collagenase P solution and digested at 37°C for about...
Embodiment 2
[0049] Embodiment 2 animal model
[0050] β-cell-specific Grb10 knockout mice Grb10KO (KO) were generated by breeding homozygous Grb10 female (loxP) mice on a C57BL / 6J background with female homozygous Ins2-Cre mice . Ins2-Cre was expressed in pancreatic β cells but not in hypothalamic neurons. loxP littermates and Ins2-Cre littermates were used as controls (WT). Ins2-Cre mice were provided by Zhang Weiping (Tianjin Medical University). All mice were housed in the Specific Pathogen Free Facility (SPF) of the Animal Health Center, Xiangya Second Hospital, Central South University, under a 12-h light-dark cycle. All procedures using animals were performed in accordance with the Animal Care and Use Committee of Xiangya Second Hospital and Central South University.
Embodiment 3
[0051] Example 3 Body Weight, Food Intake and Body Composition
[0052] Mice were fed a normal diet (ND) (Hunan SilaikeJingda Laboratory Animal Co Ltd) containing 19% protein, 5% fat and 5% fiber or a high fat drink (HFD) containing 20% protein; 60% fat; 20% carbohydrate Compound (D12492, ResearchDiets Inc.) was continued for 16 weeks. Mouse body weight and food intake were monitored weekly at the same time points. Mice were analyzed for lean and fat mass by a MQ Minispec 7.5HZ live mouse analyzer (MinispecLF50; BRUKER Optik GmbH; Germany).
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