Optic atrophy,hereditary／diagnosis; Polymerase chain reaction; DNA,mitochondrial; Point mutation; Sequence analysis
ObjectiveTo identify mutations in NDP, FZD4, LRP5, TSPAN12 in Chinese families with familial exudative vitreoretinopathy (FEVR) and observe the clinical features.MethodsRetrospective case series study. The 9 patients (18 eyes) and 5 normal members from 4 unrelated families were included in the study. The patients medical history and family history were collected in detail. All patients underwent best corrected visual acuity (BCVA), slit-lamp biomicroscopy, fundus colorized photography, fundus fluorescein angiography (FFA). Genomic DNA were collected from all the patients. Mutations were detected by directly sequencing to the whole coding region and exon-intron boundaries of NDP, FZD4, LRP5 and TSPAN12 gene. Polyphen and SIFT programs were used to predict the effects on the structure and functional properties of mutant protein.ResultsThere were two affected individuals in the family 2 carried LRP5 gene mutation [c.1330C＞T (p.R444C )] in exon 6 by sequence analysis. A score of 0.882 was acquired by Polyphen program analysis. And the missense change was predicted to be pathogenic by SIFT. Fundus changes of the proband showed angioplasia, tortuosity of peripheral vessels. And temporal dragging of the optic disc, peripheral avascular zone, neovascularization were found in FFA. Brush-like and straight of peripheral vessels were found in Ⅰ1. No variant was found in NDP, FZD4 and TSPAN12 gene.ConclusionOur study supports the gene mutation c.1330C＞T (p.R444C) of LRP5 is pathogenesis of FEVR. Patients with the same mutation could have variable phenotypic characteristics.
ObjectiveTo identify the pathogenic genes and mutations in a family with Usher syndrome type 2.MethodsA three-generation family including 7 individuals was enrolled in this study. There were 2 male patients and 5 unaffected individuals. All participants was underwent related ophthalmologic examination, including best corrected visual acuity, slit-lamp, indirect ophthalmoscopy, electroretinogram (ERG), optical coherence tomography and visual field test. DNA was extracted from 3 ml peripheral venous blood of all participants. A total of 136 hereditary retinal disease target genes were screened and the DNA sequence was performed by Next-generation sequence analysis. Then the suspected mutations compared with databases to identify the suspected mutations, which should be verified with non-affected family members and 100 normal subjects by PCR and Sanger sequence.ResultsThe sequence result showed that 2 patients, the proband and his brother, carried complex heterozygous mutations in the USH2A gene: c.5459T＞C (p.M1820T) in exon 27, c.802G＞A (p.G268R) in exon 5 and c.1190T＞A (p.I397K) in exon 7. The c.5459T＞C and c.1190T＞A mutations in USH2A have not been reported in the literature and database. Although their mother carried c.5459T＞C (p.M1820T) and c.802G＞A (p.G268R), and their father carried c.1190T＞A (p.I397K) heterozygous mutations, the parents did not present phenotype. These mutations were not detected in other normal family members. The result was supported by co-segregation analysis.ConclusionThe heterozygous mutations c.5459T＞C (p.M1820T), c.1190T＞A (p.I397K) and c.802G＞A (p.G268R) in USH2A gene cause Usher syndrome in this family.
ObjectiveTo investigate the variations in patient hospitalization expenses before the enforcement of the centralized procurement policy, after the implementation of the drug centralized procurement policy, and after the introduction of the consumables centralized procurement policy. The efficacy of the centralized procurement policy will also be examined. MethodsThis retrospective study utilizes data obtained from the medical records homepage of the Health Information Statistics Center under the Health Commission of Gansu Province. It included 32 938 inpatients who underwent PCI surgery for coronary heart disease in Gansu province between January 1, 2018, and December 31, 2022. A double-breakpoint interrupted time series model was employed to analyze the fluctuation trends in hospitalization costs among patients across various stages of the centralized procurement policy's implementation. ResultsThroughout the three phases of implementing the centralized procurement policy, the average total hospitalization costs were RMB 46 149.49 yuan, RMB 46 629.12 yuan, and RMB 28 771.76 yuan, respectively. After the centralized procurement policy with a focus on drug volume was initiated, there was an immediate reduction in average total hospitalization costs, drug costs, consumable costs, and medical service fees by 4.64%, 5.62%, 18.12%, and 8.85%, respectively. However, there was a subsequent increase of 25.28% in average medical service fees. Following this phase, average out-of-pocket costs, treatment costs, and other expenses exhibited a consistent upward trajectory, increasing by an average of 2.23%, 1.51%, and 1.21% per month. Upon the introduction of the centralized procurement policy for consumables, there was an immediate surge of 23.75% in average medical service fees, while average total hospitalization costs, out-of-pocket costs, consumable costs, treatment costs, and rehabilitation costs experienced a gradual decline. ConclusionThe enforcement of centralized procurement policies for drugs and consumables has effectively managed to reduce hospitalization costs for patients undergoing PCI surgery due to coronary heart disease, thereby easing the financial burden on patients. However, changes in consumable costs and average medical service fees were relatively modest. Going forward, it is essential to refine the centralized procurement policy concerning consumables, improve the compensation mechanism for medical service pricing, and enhance the overall value proposition of medical services.
ObjectiveTo reveal the pathogenic mutation in a three-generation Chinese family with autosomal dominant familial exudative vitreoretinopathy (FEVR). MethodsThree patients and a healthy spouse from the index family with FEVR were recruited. The proband was a 5 years old boy. His mother and grandpa were presented with typical FEVR presentations, while his father with normal ocular fundus. DNA was extracted from peripheral blood samples taken from all four participants. All coding and exon-intron boundary regions of five targeted genes, including NDP, FZD4, LRP5, TSPAN12 and ZNF408 were amplified with polymerase chain reaction and sequenced using direct sequencing. In silico analyses were applied to determine the conservation of the mutation site, pathogenic effect and the potential protein crystal structural changes caused by the mutation. ResultsFZD4 c.478G > A, a susceptible mutation was found after four high frequency mutation sites which MAF values were higher than 0.001 was filtered among 5 single nucleotide variations detected in four participants, leading to the residue 160 changing from glutamate to lysine (p.E160K). Co-segregation analysis between genotypes and phenotypes revealed FZD4 p.E160K as the disease-causing mutation for this family. Conservational analysis suggested that this mutation site was highly conserved among all tested species. Functional analysis predicated that this mutation may be a damaging mutation. Crystal structural analysis also indicated that this mutation could lead to the elimination of the hydrogen bond between residue 160 and asparagine at residue 152, thus altering the tertiary structure of the protein and further impairing the protein function. ConclusionOur study demonstrates FZD4 p.E160K as a novel pathogenic mutation for FEVR.
Interrupted time series (ITS) analysis is a quasi-experimental design for evaluating the effectiveness of health interventions. By controlling the time trend before the intervention, ITS is often used to estimate the level change and slope change after the intervention. However, the traditional ITS modeling strategy might indicate aggregation bias when the data was collected from different clusters. This study introduced two advanced ITS methods of handling hierarchical data to provide the methodology framework for population-level health intervention evaluation.
ObjectiveTo identify the pathogenic mutation in a three generation Chinese family with low penetrance retinoblastoma (RB). Methods8 from 9 family members received complete ophthalmic examinations. DNA was extracted from 6 family members. Using exon combined target region capture sequencing chip to screen the candidate disease-causing mutations. Sanger sequencing were used to confirm the disease-causing mutation. ResultsAmong 9 family members, the proband (Ⅲ2) was bilateral RB, Ⅲ1 was unilateral RB, Ⅲ3 was dead for bilateral RB. Normal fundus were observed in the left eye ofⅢ1 and the eyes of other family members except the proband. Sequence analysis of RB1 gene revealed a missense mutation c.1981C > T (p.Arg661Trp) in the proband and two carriers (Ⅱ2, Ⅱ3), but not in the two normal subjects (Ⅱ1, Ⅱ4). We suspect that the RB penetrance in the family was 50%. ConclusionsThere is a missense mutation c.1981C > T in a Chinese family with low penetrance RB. The RB penetrance is 50%.
Objective Molecular cloning of rat retinal degeneration slow(RDS)gene cDNA. Methods Using PolyA+RNA from retina of SD rat as template,a 1555bp positive cDNA band was obtained by RT-PCR and subcloned into pBluescriptⅡKS(+) vector.The cloned fragment was analyzed with restriction endonucleases and sequencing. Results It had been proved that the cloned fragment was rat RDS/peripherin cDNA.Except for the substitute of A1242G and CA1409-1411CCA,the other sequences corresponded to that reported by Begy. Conclusion Rat RDS/peripherin cDNA was obtained.Researches on function of rat RDS/peripherin gene and its role in retinal degeneration are under way. (Chin J Ocul Fundus Dis,1999,15:97-99)
PURPOSE:To investigate the status and detailed structure of Rb gene in primary tumors and somatic cells of patients with retinoblastoma. To identify the character, origin and transmission of oncogenie point mutations. METHODS:DNA hybridization,SSCP analysis and PCR-associated direct sequencing. RESULTS:Among 108 RB patients examined 80 cases were found to have subtle alterations affecting Rb locus,including 44 cases with homozygous Rb point mutations, 20 cases with two independent heterozygous Rb point mutations, 16 cases with heterozygous mutations involved in one allele of Rb gene. Majority of bilateral RB patients and a small fraction of unilateral RB patients were detected to have a germline mutation. In addition the higher frequency of new germline mutation and parental origin of mutation were observed. CONCLUSION :Rb gene is closely associated with retinoblastoma. Two mutation events and resulting inaetivations of both Rb alleles are required for RB tumorigenesis. Based on our own data,the first event is exclusively point mutation. As for the second event,LOH accounts for two third of cases,point mutation for one third of cases. （Chin J Ocul Fundus Dis,1997,13: 12- 16）
Purpose To identify the expression of alternatively spliced mRNA isoforms of the NMDA-R1 in the visual cortex of strabismic cats. Methods Two pai rs of normal and strabismic cats were used．The amblyopic cats had been made monocularly esotropic (by tenotomy) at the age of weeks，resulting in behavioral am blyopia．Animals were sacrificed about 6 months by intraperitoneal administration of Nembutal．Cryostat sections of fresh，frozen central visual cortex of the ats were cut to 20 micron thickness．A series of digoxygenin-labelled oligonucle otide probes basing on the human gene sequence were used for ISH．Control probes included sense oligonucleotides and short segment probes which were adjacent to ，but did not，span the splice junctions．A computer-assisted systematic morphometric ounting procedure was used to enumerate hybridising cells． Results The number of positive cells expressing NMDA-R1 mRNA in t he strabismic amblyopic cats was decreased，notably in layer IV of visual cortex (P＜0．0001)．The pattern of isoform expression varied between normal and strabismic amblyopic cats with decreased numbers of 1-a，1- b and 1-1 isoforms and apparently increased expression of 1-3 P ＜0．0001)，whereas no significant difference was found for the 1-2 and 1-4 isoforms (P＞0．05)． Conclusion Transcriptional inhibition of NMDA-R1 mRNA and of specifie isoforms may underlie the change in receptor expression．Alternatively，preferentialloss of neurones bearing particular NMDA-R1 isoforms and compensation with a proportional increase in cells expressing other isoforms may occurr during the critical period of visual plasticity． (Chin J Ocul Fundus Dis，2000，16：71-138)