Objective To set up the model of deiodinase ( Dio) 3b-/- zebrafish and to observe the effect of which on embryo development. Methods The zebrafish model of Dio3b-/- was set up by CRISPR-Cas9 gene editing technology, PCR and sequencing was used to confirm the efficiency of deletion. The heart rate of embryos at 48 hours post fertilization was counted. The locomotor activity of 5-7 days post fertilization larve was detected using behavior tracking system. Results The model of Dio3b knockout zebrafish was set up successfully. The heart rate of embryos Dio3b-/- increased ( P＜0. 001) and the locomotor activity of 5-7 days post fertilization larves lacking Dio3b gene increased (P＜0.05) significantly compared with that of wild type control respectively. Conclusion The deletion of zebrafish Dio3b gene results in the phenotype of hyperthyroidism and the model of Dio3b-/- is proper for studying the effect of partial excess thyroid hormone on embryo development.

Objective To investigate the prevalence of DUOX2 mutations in Chinese patients with congenital hypothyroidism (CH) and to discuss the inheritance pattern of DUOX2 gene.Methods Blood samples were collected from 91 CH children and their genomic DNA was extracted from peripheral blood leukocytes.All exons and exon-intron boundaries of DUOX2 were analyzed by target next-generation sequencing and family trios was established to study the inheritance pattern of DUOX2 gene.Results Fifty-four out of 91 children with CH carried DUOX2 mutation, with a prevalence of 59.34%.Of the 54 CH children, 36 carried DUOX2 biallelic mutations.In all 12 family trios with probands carrying biallelic DUOX2 mutations, the parents carried heterozygous DUOX2 mutations while still showing normal thyroid function, suggesting that CH caused by DUOX2 mutations is inherited in an autosomal recessive manner.Conclusion DUOX2 gene is one of the most frequently mutated genes in Chinese CH patients and its inheritance pattern is an autosomal recessive one.

Objective:To assess clinical and genetic features in a patient with thyroid hormone resistance syndrome(RTH) and explore the pathogenic mechanism.Methods:The clinical data of the proband was collected. The genomic DNA was extracted from peripheral blood samples of the patients. The pathogenic variant was identified using whole-exome sequencing and confirmed by Sanger sequencing. Then the function of the mutation sites was detected by bioinformatics.Results:The patient presented with chest distress, palpitation, and persistent atrial fibrillation, along with elevated levels of serum free triiodothyronine(FT 3), free thyroxine(FT 4), and thyroid stimulating hormone(TSH), which suggested RTH clinically. The genetic analysis identified a heterozygous mutant of THRβ(c.1313G>A) gene at exon 8, which was a missense mutation causing the substitution of arginine to histidine at 438 position of the protein(p.R438H). Its inheritance pattern was unknown. This mutation was considered as a new one that had not been reported. Conclusion:A novel pathogenic THRβ gene mutation was found in the patient with RTH, which might be the cause of this disease. This variant c. 1313G>A is located in the ligand binding domain of THRβ, which might result in low protein activity.

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

ABSTRACT OBJECTIVE：To investigate the quality differences of Pinelliae Rhizoma with different diameter，and to providereference for the rational utilization of its resources. METHODS：A total of 65 batches of commercial Pinelliae Rhizoma from thefour major medicinal markets in China and two companies were collected as samples，the diameter of them were measured byvernier caliper，and then them were screened into samples with different diameter ranges（0.5 cm≤d≤0.8 cm，0.8 cm＜d≤1.0cm，1.0 cm＜d≤1.2 cm，1.2 cm＜d≤1.5 cm，1.5 cm＜d≤2.0 cm）by sieving. The content of organic acids（oxalic acid，citricacid，L-malic acid，succinic acid，fumaric acid，trans-aconitic acid and cis-aconitic acid）in Pinelliae Rhizoma samples of differentdiameter ranges were determined by HPLC，and the content of the extracts of Pinelliae Rhizoma samples of different diameterranges were determined by cold leaching method according to 2201 general principles in 2015 edition of Chinese Pharmacopoeia（Part Ⅳ），to evaluate the quality difference of Pinelliae Rhizoma in different diameter ranges.73 batches of Pinelliae Rhizoma fromcorresponding 10 main producing areas were collected as samples，to investigate its distribution of diameter. RESULTS：Dtermination of 65 batches of commercial Pinelliae Rhizoma showed that the content of organic acids and extracts in PinelliaeRhizoma increased with the increase of its diameter in general. However，there were no significant difference in organic acids andcontent of the extracts among Pinelliae Rhizoma samples of different diameter ranges in the same batch（P＞0.05）. The diameter distribution of 73 batches of Pinelliae Rhizoma from corresponding areas was mainly between 0.6 cm and 1.8 cm，with an averageweight of 95.54％. If the diameter standard（1-1.5 cm）specified in the 2015 edition of Chinese Pharmacopoeia（part Ⅰ）wasadopted，only 8 of 73 batches of Pinelliae Rhizoma meet the requirements，and the qualified rate was 10.96％；if the diameterrange was properly expanded to 0.7-1.5 cm，54 batches of Pinelliae Rhizoma meet the requirements，with a qualified rate of73.97％；when the diameter range was expanded to 0.7-1.6 cm，68 batches of Pinelliae Rhizoma meet the requirements，with aqualified rate of 93.15％ .CONCLUSIONS：There is no significant difference in the quality of Pinelliae Rhizoma with different diameters. Combined with the diameter distribution of Pinellia Rhizoma in different producing areas，the diameter range of 1-1.5cm specified in relative standard can be expanded to 0.7-1.6 cm，avoiding the waste of resources.