Objective To analyze and identify the mutations in SGLT2 gene of nine Chinese families with FRG, and determine the renal threshold for glucose excretion (RTG), so as to explore the association of genotype and RTG. Methods All coding regions of SGLT2 gene, including intron exon boundaries, were analyzed using PCR followed by direct sequence analysis. Quantitative test for 24?hour urine glucose and RTG were measured among 9 probands (21 patients) and their family members from 9 pedigrees (total 25 subjects). The differences in renal glucose thresholds between patients with different genotypes (heterozygotes and compound heterozygotes; c.886(-10_-31) del heterozygotes and other heterozygotes) were compared. Results Twelve mutations were identified by SGLT2 gene analysis, including 10 novel ones that were not included in HGMD:c.331T>C, p.W111R;c.374T>C, p.M125T; c.394C>T, p.R132C; c.612G>C, p.Q204H; c.829C>T, p.P277S; c.880G>A, p.D294N;c.1129G>A, p.G377S; c.1194C>A, p.F398L; c.1540C>T, p.P514S; c.1573C>T, p.H525Y. In thisstudy, the mutation c.886(-10_-31)del that is specific to Chinese population accounted for about 28％of the total alleles (5/18). The RTG values of patients with compound heterozygous mutations were much lower than those with simple heterozygous mutations [(1.28 ±0.10) vs (5.14±0.77) mmol/L; P<0.001];and c.886(-10_-31)del heterozygotes had significant lower RTG values than others [(4.43 ± 0.37) vs (5.70 ± 0.51) mmol/L, P<0.001]. Conclusions Ten novel mutations which may be related to FRG are found in this study, and c.886(-10-31)del may be a hot?spot mutation in Chinese patients. Compound heterozygotes had much lower RTG values than simple heterozygotes.

OBJECTIVE To isolate and identify the chemical compounds of the n-butanol fraction of the pods of Macleaya microcarpa, and to predict the targets and pathways for its treatment of Alzheimer's disease by network pharmacology. METHODS Silica gel, Sephadex LH-20 column chromatography and ODS preparative liquid chromatography were used to separate and purify the n-butanol extract of the pods of Macleaya microcarpa, and the structure of the compounds were identified by NMR spectroscopic. The targets of active compounds were obtained using SwissTargetPrediction and Targetnet database. Alzheimer's disease-related target were obtained by Genecard, OMIM and TTD database. The two were imported into Venny online tool to select the effect targets of the active compounds for Alzheimer's disease treatment. The interaction network map of “drug-component-target-disease” was constructed by Cytoscape 3.9.1 software. The String database was used to build the protein protein interaction network analysis. KEGG and GO pathway enrichment analysis were performed with Metascape database, and molecular docking was studied with AutoDockTools 1.5.7 software. RESULTS Five phenolic glycosides were isolated from the n-butanol fraction of the pods of Macleaya microcarpa. Ten key targets such as TNF, PTGS2 and APP, and 10 important pathways such as Pathways in cancer, Serotonergic synapse and Alzheimer's disease were screened by network pharmacology, and molecular docking showed that the active ingredients had good binding ability to the key targets. CONCLUSION Compounds 3-5 are isolated from Macleaya microcarpa for the first time, and compound 5 is isolated from Papaveraceae for the first time. The mechanism of Macleaya microcarpa in the treatment Alzheimer's disease may affect on key targets such as TNF, PTGS2, APP, ABCB1, and influence signaling pathway such as the APP/Aβ/NMDAR, so as to reduce inflammatory factors, inhibit inflammatory responses and reduce Aβ deposition in the brain.