Objective:To investigate and analyze the relationship between genotype and phenotype in patients with early-onset high myopia (eoHM) associated with hereditary eye diseases.Methods:A family-based study was conducted among 30 families diagnosed with eoHM at Department of Ophthalmology of People's Hospital of Ningxia Hui Autonomous Region from January 2022 to June 2023. Seven families (23.3%, 7/30), all probands, and their 14 parents were included. These seven families were unrelated. Detailed patient and family histories were collected. All participants underwent comprehensive ophthalmic examinations, including best-corrected visual acuity, color vision testing, fundus color photography, optical coherence tomography, fluorescein fundus angiography, and fundus autofluorescence imaging. Full-field electroretinography was performed in four cases. Peripheral venous blood samples were collected from patients and their parents for whole-genome DNA extraction and whole-exome sequencing. Potential pathogenic variants were identified, and their pathogenicity was analyzed and confirmed by Sanger sequencing. The pathogenicity of newly discovered gene variants was evaluated according to the guidelines of the American College of Medical Genetics and Genomics (ACMG). Literature on previously reported eoHM associated with hereditary eye diseases was reviewed to analyze the relationship between variant genes and clinical phenotypes.Results:Among the seven families, three exhibited X-linked inheritance, two showed autosomal recessive inheritance, and two demonstrated autosomal dominant inheritance. All the patients were male. Among the seven patients, one case each was identified with congenital stationary night blindness (CSNB), Bornholm eye disease, X-linked retinitis pigmentosa (XL-RP), cone-rod dystrophy, Knobloch syndrome, familial exudative vitreoretinopathy (FEVR), and Stickler syndrome. Genetic testing revealed nine gene variants highly correlated with the observed phenotypes. The genetic testing results revealed that all patients were found to carry nine gene variants highly associated with the phenotype, including: a hemizygous missense variant NYX c.647A>T (p.N216I) (M1), an OPN1LW LIAVA haplotype variant (M2), a hemizygous frameshift variant RPGR c.3096_3097del (p.E1033RfsTer45) (M3), compound heterozygous variants TTLL5 c.1588_1589del (p.L531EfsTer24) and c.850G>C (p.D284H) (M4, M5), compound heterozygous variants COL18A1 c.2118dup (p.G707RfsTer23) and c.3523_3524del (p.L1175VfsTer72) (M6, M7), a heterozygous missense mutation FZD4 c.1499C>T (p.T500I) (M8), and a heterozygous frameshift variant COL11A2 c.966dup (p.T323HfsTer19) (M9). Among them, M2, M4, M5, M8 and M9 were newly discovered mutation sites, and M1, M3, M6 and M7 were known mutation sites. According to the classification standards and guidelines of genetic variation issued by ACMG, M2, M3, M4, M6, M7, and M9 were judged to be pathogenic variants, while M1, M5, and M8 were of unknown clinical significance. Through literature review, it was found that eoHM was more common among the clinical phenotypes of 4 types of hereditary retinal diseases, including CSNB, Stickler syndrome, FEVR and XL-RP. Conclusion:eoHM is intricately associated with inherited eye diseases and may serve as the earliest indicator of such conditions.

Objective:To investigate and analyze the relationship between genotype and phenotype in patients with early-onset high myopia (eoHM) associated with hereditary eye diseases.Methods:A family-based study was conducted among 30 families diagnosed with eoHM at Department of Ophthalmology of People's Hospital of Ningxia Hui Autonomous Region from January 2022 to June 2023. Seven families (23.3%, 7/30), all probands, and their 14 parents were included. These seven families were unrelated. Detailed patient and family histories were collected. All participants underwent comprehensive ophthalmic examinations, including best-corrected visual acuity, color vision testing, fundus color photography, optical coherence tomography, fluorescein fundus angiography, and fundus autofluorescence imaging. Full-field electroretinography was performed in four cases. Peripheral venous blood samples were collected from patients and their parents for whole-genome DNA extraction and whole-exome sequencing. Potential pathogenic variants were identified, and their pathogenicity was analyzed and confirmed by Sanger sequencing. The pathogenicity of newly discovered gene variants was evaluated according to the guidelines of the American College of Medical Genetics and Genomics (ACMG). Literature on previously reported eoHM associated with hereditary eye diseases was reviewed to analyze the relationship between variant genes and clinical phenotypes.Results:Among the seven families, three exhibited X-linked inheritance, two showed autosomal recessive inheritance, and two demonstrated autosomal dominant inheritance. All the patients were male. Among the seven patients, one case each was identified with congenital stationary night blindness (CSNB), Bornholm eye disease, X-linked retinitis pigmentosa (XL-RP), cone-rod dystrophy, Knobloch syndrome, familial exudative vitreoretinopathy (FEVR), and Stickler syndrome. Genetic testing revealed nine gene variants highly correlated with the observed phenotypes. The genetic testing results revealed that all patients were found to carry nine gene variants highly associated with the phenotype, including: a hemizygous missense variant NYX c.647A>T (p.N216I) (M1), an OPN1LW LIAVA haplotype variant (M2), a hemizygous frameshift variant RPGR c.3096_3097del (p.E1033RfsTer45) (M3), compound heterozygous variants TTLL5 c.1588_1589del (p.L531EfsTer24) and c.850G>C (p.D284H) (M4, M5), compound heterozygous variants COL18A1 c.2118dup (p.G707RfsTer23) and c.3523_3524del (p.L1175VfsTer72) (M6, M7), a heterozygous missense mutation FZD4 c.1499C>T (p.T500I) (M8), and a heterozygous frameshift variant COL11A2 c.966dup (p.T323HfsTer19) (M9). Among them, M2, M4, M5, M8 and M9 were newly discovered mutation sites, and M1, M3, M6 and M7 were known mutation sites. According to the classification standards and guidelines of genetic variation issued by ACMG, M2, M3, M4, M6, M7, and M9 were judged to be pathogenic variants, while M1, M5, and M8 were of unknown clinical significance. Through literature review, it was found that eoHM was more common among the clinical phenotypes of 4 types of hereditary retinal diseases, including CSNB, Stickler syndrome, FEVR and XL-RP. Conclusion:eoHM is intricately associated with inherited eye diseases and may serve as the earliest indicator of such conditions.

Objective To investigate the mechanism via which artesunate regulates the invasion and metastasis of colon cancer cells and the expression of members of the TGF-β1/Smad4 signaling pathway. Methods The cell counting kit 8 (CCK8), nude mouse xenograft model, Transwell invasion assay, and flow cytometry were used to investigate the effect of artesunate on the invasion and metastasis of colon cancer cells. Western blotting and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the expression of TGF-β1 and Smad4 proteins and mRNA, respectively. Results Artesunate inhibited the growth of transplanted tumor, cell proliferation, and invasion and promoted apoptosis. It inhibited TGF-β1 expression and promoted Smad4 expression. TGF-β1 inhibitors reversed the inhibitory effect of artesunate. Conclusion Artesunate can inhibit the growth of xenograft tumor in nude mice and its mode of action may be related to the TGF-β1/Smad4 signaling pathway.

Objective To investigate the effect oftopiramate on NACHT-LRR-PYD-containing protein 3 (NALP3) inflammasome and intedeukin (IL)-1β levels in trigeminal ganglion of migraine rats.Methods Forty adult male Sprague Dawley rats were randomly divided into blank group,saline group,model group,prevention control group,10 mg/kg topiramate group,30 mg/kg topiramate group,60 mg/kg topiramate group,and 90 mg/kg topiramate group (n=5).Inflammatory soup was used to stimulate the dual matter of rats repeatedly to induce migraine models:rats in the blank group were without any treatment,those in the saline group were given saline to stimulate the dual matter,different concentrations of topiramate group were given to migraine models of the 10 mg/kg topiramate group,30 mg/kg topiramate group,60 mg/kg topiramate group,and 90 mg/kg topiramate group,respectively,and migraine models of the prevention control group were given saline containing 1％ Tween80.Three h after the last treatment,the expressions of NALP3,caspase-1 precursor (pro-caspase-1),caspase-1 and IL-1β in trigeminal ganglion of rats were detected by Western blotting,and the best concentration oftopiramate was chosen for subsequent immunofluorescence experiments.Ten healthy male adult SD rats were randomly divided into control group and topiramate group (n=5);the expression levels of NALP3,caspase-1 and IL-1β in trigeminal ganglion of the two groups were detected by immunofluorescence.Results The expression levels ofNALP3,pro-caspase-1,caspase-1 and IL-1β were not significantly different between saline group and blank group (P＞0.05);the expression levels ofNALP3,pro-caspase-1,caspase-1 and IL-1β in the model group were significantly higher than those in the saline group (P＜0.05).The expression levels of NALP3,pro-caspase-1,caspase-1 and IL-1β in 60 mg/kg topiramate group and 90 mg/kg topiramate group were significantly lower than those in the prevention control group (P＜0.05).The fluorescence intensity ofNALP3,caspase-1 and IL-1β in topiramate group was significantly lower than that in control group.Condusion Topiramate can inhibit the expression of NALP3 inflammasome and IL-1β in the trigeminal ganglion of migraine rat models,and it is likely to be one of the important mechanisms for the prevention of migraine.

OBJECTIVE:To explore the association between the mitochondrial DNA(mtDNA)3537A/G,5351A/G variant and type 2diabetes mellitus(T2DM)in northem Chinese population.METHODS:The subjects including 614 patients with T2DM in Dalian City,61 of them were collected from family survey,497 of them were collected from Department of Endocrine,Dalian Municipal Central Hospital,and the remained 56 were selected from diverging T2DM patients in Dalian City.Additional 344 cases with normal carbohydrate toierance were served as controls.The mtDNA 3537A/G.5351A/G variants in 614 patients with T2DM and 334 healthy control subjects were examined.By sequencing the mtDNA in 24 cases and 26 controls,2 candidate SNPs in mtDNA were determined,and then genotyping was carried out by using PCR restriction fragment length polymorphism(PCR-RFLP)analysis.RESULTS:The frequency of mtDNA A3537G and A5351G mutation was 2.0%and 2.6% in T2DM patients,respectively,which was 2.1%and 4.2% in the control group.No significant difference had been observed between case and control(P＞0.05).After stratifying by body mass index and blood pressure,we found that the frequency of A5351 G in obesity patients with T2DM was 1.61%,and in obesity control was 15.38%,which had significant difference(P_(Fisher)=0.02,OR=2.76),however,A3537G stili showed no significant difference in all groups(P＞0.05).CONCLUSION:5351A/G in mtDNA ND2 gene may be a variance associated with T2DM in northem Chinese.