Nucleus plasma ratio was measured and silver-binding nucleolar organizer (AgNORs) were counted in 31 cases of cholangiocarinoma (11 cases were well-differentiated, 10 case moderately differentiated and 10 cases poorly differentiated) and in 17 cases of atypical epithelial hyperplasia related to hepatolithiasis (9 cases were simple hyperplasia, 8 cases atypical epithelial hyperplasia) by AgNORs techique and image analysis.The results showed that mucleus plasma ratio and AgNORs counts increased significantly from well-differentiated to poorly differentiated cholangiocarcinoma (Ｐ＜0.01). No statistically significant differance was shown between nucleus plasma ratio of atypical hyperplasia and well-differentiated cholangiocarinoma.The data imply that chronic proliferative cholngitis in the presence of hepatolithiasis can progress to atypical epithelial hyperplasia which may be an important precursor of cholangiocarcinoma.
ObjectiveTo investigate the clinical risk factors of preterm infants and its severity in premature infants with bronchopulmonary dysplasia (BPD) with retinopathy of prematurity (ROP).MethodsRetrospective clinical study was performed. A total of 126 preterm infants with BPD in the Neonatal Department of the Affiliated Hospital of Qingdao University from January 2016 to December 2018 were enrolled in the study. Among them, 69 were males and 57 were females, whose gestational age＜32 weeks and birth weight＜1500 g. BPD grades Ⅰ, Ⅱ, and Ⅲ were 63, 40, and 23 cases respectively. According to the presence or absence of ROP, children were divided into ROP group and non-ROP group, with 48 (38.1%) and 78 (61.9%) cases respectively. The differences of clinical data between the two groups were compared and analyzed. Quantitative data comparison between groups was performed by t test, and count data comparison was performed by χ2 test. The risk factors of ROP in BPD premature infants were analyzed by multi-factor logistics regression. The correlation between BPD severity and ROP severity was tested by Spearman rank correlation test.ResultsCompared with the non-ROP group, the ROP group had a smaller gestational age (t=5.988), lower birth weight (t=7.371), higher the application rate of oxygen concentration＞30% (duration of service＞24 h), high rate (χ2=17.244) and longer noninvasive ventilation time (t=－7.139), the differences were statistically significant (P＜0.05). In the logistic regression model, the noninvasive ventilation time was the risk factor for ROP in preterm infants with BPD (OR≈1.054, P＜0.05), while gestational age and birth weight were importantly protective factors for ROP in preterm infants with BPD (OR≈0.938, 0.996; P＜0.05). The results of the correlation analysis found that the severity of BPD was significantly positively correlated with the severity of ROP. As the severity of BPD increased, the severity of ROP increased, and the difference was statistically significant (rs=0.306, P＜0.035).ConclusionsFetal gestational age, low birth weight, hyperoxia, and long-term non-invasive mechanical ventilation are the main risk factors for ROP in preterm infants with BPD. The severity of BPD is positively correlated with the incidence and severity of ROP.
ObjectiveTo observe and analyze the clinical phenotype and genetic characteristics of COL2A1 and COL11A1 de novo mutation (DNM) related Stickler syndrome type Ⅰ and Ⅱ patients. MethodsA family-based cohort study. From December 2023 to November 2024, 4 patients (all probands) with Stickler syndrome diagnosed by clinical and genetic testing in Department of Ophthalmology of People's Hospital of Ningxia Hui Autonomous Region and their parents (8 cases) were included in the study. The patients came from 4 unrelated families. A detailed medical history was taken, and the patients underwent best-corrected visual acuity (BCVA), refraction, and fundus color photography examinations. Systemic examinations included the oral and facial regions, skeletal, joints, and hearing. Peripheral venous blood samples were collected from the patients and their parents, and genomic DNA was extracted. Whole-exome sequencing was used to screen for pathogenic genes and their loci, which were then validated by Sanger sequencing and combined with segregation analysis in the families to identify candidate gene mutation sites. The candidate variants were assessed for pathogenicity according to the American College of Medical Genetics and Genomics (ACMG) criteria and guidelines for the classification of genetic variants. Additionally, cross-species conservation analysis was performed to determine the evolutionary conservation of wild-type amino acids, and protein three-dimensional modeling techniques were used to characterize the spatial conformational changes of the variant proteins and the alterations in their local hydrogen bond networks. ResultsAmong the 4 patients, there were 2 males and 2 females; their ages ranged from 3 to 12 years. There were 2 cases of Stickler syndrome type Ⅰ (proband of families 1 and 2) and 2 cases of type Ⅱ (proband of families 3 and 4). The diopters ranged from −8.00 to−18.00 D. BCVA ranged from no light perception to 0.6-. There were 2 cases each of vitreous membrane-like and “bead-like” opacity. Three cases showed peripapillary atrophy arcs and leopard pattern changes in the retina; one case had bilateral retinal detachment with a large macular hole in the left eye, which had previously been treated with vitrectomy surgery. One case had bilateral sensorineural hearing loss. There were 3 cases of simple micrognathia; one case had a flat nasal bridge, short nose, midface depression, and micrognathia. Two cases had excessive elbow joint extension. The phenotypes of the parents of the 4 patients were normal. Genetic testing results revealed that the probands of families 1 and 2 carried COL2A1 gene c.85+1G>C (M1) splice site variant and c.3950_3951insA (p.M1317Ifs*48) (M2) frameshift variant, respectively; the probands of families 3 and 4 carried COL11A1 gene (NM_001854.4) c.2549 G>T (p.G850V) (M3) missense variant and c.3816+6T>C (M4) splice site variant, respectively. The parents did not carry the related gene variants. Among them, M2, M3, and M4 are newly reported DNM. According to the ACMG guidelines, they were all considered likely pathogenic. The cross-species conservation analysis results showed that the wild-type amino acid of the COL11A1 gene M3 missense variant was highly conserved across multiple different species. Protein local structure modeling analysis revealed that the COL2A1 gene M2 frameshift variant and the COL11A1 gene M3 missense variant significantly altered the tertiary structure conformation of the protein, leading to abnormal spatial arrangement and hydrogen bond network in the key functional domains ConclusionThe COL2A1 gene M1 splice site variant, M2 frameshift variant, and the COL11A1 gene M3 missense variant, M4 splice site variant are respectively the potential pathogenic genes for families 1, 2, and families 3, 4; leading to the onset of Stickler syndrome type Ⅰ in families 1 and 2, and type Ⅱ in families 3 and 4.
ObjectiveTo investigate and analyze the relationship between genotype and phenotype in patients with early-onset high myopia (eoHM) associated with hereditary eye diseases. MethodsA 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. ResultsAmong 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. ConclusioneoHM is intricately associated with inherited eye diseases and may serve as the earliest indicator of such conditions.