Rapid prediction of G protein-coupled receptor structures using nanoluciferase assay / 药学学报

Using beta-2 adrenergic receptor, 5-hydroxytryptamine and angiotensin II type 1 receptor as control, we here established a method for rapid prediction of the initial position amino acids of N-terminal, C-terminal, intracellular loops, extracellular loops and transmembrane (TM) regions in G protein-coupled receptors (GPCRs), and successfully predicted the structure of Mas-related G protein-coupled receptors X3 (MRGPRX3). To achieve this purpose, nanoluciferase (Nluc) was inserted into the different sites of these GPCRs′ sequence by sequence and ligation-independent cloning (SLIC) method, and the luminescence value were measured to distinguish the different parts of GPCRs. The results showed that luminescence values of NLuc luciferase at TM region were less than 100 000, and the values were higher than 1 000 000 at N terminal, C terminal, or extracellular loops and intracellular loops, and the values were between 100 000 and 500 000 at junction. The predicted MRGPRX3 structure was analyzed in detail and was compared with AlphaFold predicted structure. In conclusion, this method could provide useful information of GPCR structure model for the ligand virtual screening, and could provide certain experimental basis for structural pharmacology.

Clinical Efficacy of Modified Sinisan Combined with Saccharomyces Brasiliensis on Functional Dyspepsia in Children and Its Effect on Serum Cytokines / 中国实验方剂学杂志

Objective: To discuss the clinical efficacy of modified Sinisan (Qizhi Weitong granule) and saccharomyces boulardi in treating functional dyspepsia (FD) in children, and its effect on changes in serum cytokine and gastrointestinal hormones.Method: Totally 156 cases were divided into control group (78 cases) and observation group (78 cases). The control group was treated with saccharomyces boulardi, 0.25 g, po, bid, after meal. In addition to the therapy of control group, the observation group was also given modified Sinisan (Qizhi Weitong granule), 2.5 g, po, tid, 30 minutes before meal. Four weeks was a course of treatment. Then the clinical efficacy, the time of symptom improvement and the change of serum cytokine and gastrointestinal hormones were observed.Result: The total effective rate of observation group (96.15%) was higher than that of control group (82.05%), with statistical difference (Pγ(IFN-γ), tumor necrosis factor-α(TNF-α), plasma motilin(MTL) and Leptin of both groups had no statistical difference before treatment. After treatment, the serum levels of IL-10, MTL of both group were increased, while the serum levels of IFN-γ, TNF-α, Leptin of both group were decreased. However, the decreasing range and the increasing range of observation group were statistically obvious than those of control group (PConclusion: Modified Sinisan(Qizhi Weitong granule) combined with saccharomyces boulardi have an obvious clinical efficacy in treating child's FD, with a low recurrence. The mechanism may be correlated with the effect of integrated Chinese and Western medicines in regulating serum cytokine and gastrointestinal hormones levels.

Change of peripheral blood appetite regulation factor of anorexia children and infect of child anorexia granule / 中国中药杂志

Study the infect of child anorexia granule on serum ghrelin and leptin of anorexia children and its clinical efficacy. Selected 81 cases of anorexia children aged 1-6 years old into treatment group (42 cases) and control group (39 cases), in addition, 30 case healthy children as healthy control group. The control group children were treated with domperidone suspension 0.3 mg x kg(-1) x d(-1), tid, orally 30 minutes before meals. Treatment group were treated with child anorexia granule, 1-3 years 1 package, bid; 4-6 years 1 package, tid; po, 4 weeks as a course of treatment. Study the change of serum ghrelin and leptin before and after therapy. The study demonstrates that before treatment, the serum ghrelin level of disease group was lower than healthy group (P < 0.01), and the serum leptin level was higher than healthy group (P < 0.01). After treatment, the serum ghrelin level both increase, and the serum leptin decline. And the change of treatment group was significantly different with control group (P < 0.01). And the clinical effective rate are 95.23% and 74.35% (P < 0.01). After 6 months of follow-up visit, the children weight significantly increase in treatment group (P < 0.01). Results indicate that child anorexia granule can facilitate secretion of ghrelin, and inhibit secretion of leptin, so as to work up an appetite. And the molecular mechanism is its infect on serum ghrelin, leptin.