Search

Expressions of apoptosis-related genes of retinal blood vessel in early diabetic rats

Objective To analyze the expression of apoptosis-related genes of retinal blood vessel in early diabetic rats by gene chip technology. Methods To make diabetic rat model by intraperitoneal injection of streptozotocin (STZ). On the 6th week after blood pressure increased, 10 rats were executed in Diabetic group and normal control group respectively. 20 retinal blood vessels were extracted and the RNA was isolated. The probe was made of alpha;-32 P-deoxyadenosine triphosphate (dATP)-labeled sample which hybridized 1176 nylon chips, and then analyzed by software. Three different expression genes were selected to verify by reverse transcription polymerase chain reaction (RT-PCR). Results On the 6th week, 136 （11.5％） genes were differentially expressed ［up-regulated genes were 90(7.6%), down-regulated genes were 46(3.9%)］in diabetic group. These genes involved into different groups according to their function. Especially in 72 apoptosis-related genes, 15 genes were differentially expressed. The up-regulated genes were some TNF receptor family members such as TNFRSF12, TRAIL, TNFRSF9, FADD；Bcl-2 family members such as bcl-w, bax, bak1 and AKT. The down-regulated genes were FAF1 which related to fas. Conclusions The expression of retinal vascular gene in early diabetic rats has been changed complicatedly. In particular, the multiple apoptosis-related genes have been changed in early diabetic, and most of them are at the upstream of apoptosis pathway. These findings indicate that the development of diabetic retinopathy is associated with multiple signaling pathways leading to apoptosis, while the alterations on the level of molecular biochemistry are still limited in apoptosis induction period. （Chin J Ocul Fundus Dis，2008，24：244-248）

抗人视网膜母细胞瘤单克隆抗体轻链可变区基因克隆、测序

Analysis of mutations of disease-causing gene and the inheritance in sporadic retinitis pigmentosa

ObjectiveTo observe the disease-causing genes and the inheritance in sporadic retinitis pigmentosa (sRP) in Ningxia region. Methods49 sRP patients and 128 family members were recruited for this study. All the patients and family members received complete ophthalmic examinations including best corrected visual acuity, slit-lamp microscope, indirect ophthalmoscopy, fundus color photography, visual field, optic coherence tomography, full view electroretinogram. DNA was extracted from patients and family members. Using exon combined target region capture sequencing chip to screen the 230 candidate disease-causing gene mutations, polymerase chain reaction and direct sequencing were used to confirm the disease-causing mutations. Results24/49 patients (49.0%) had identified disease-causing genes, totally 16 genes were involved. There were 41 mutation sites were found, including 32 new mutations (78.0%). The disease-causing genes include USH2A, C2orf71, GNGA1, RPGR1, IFT140, TULP1, CLRN1, RPE65, ABCA4, GUCA1, EYS, CYP4V2, GPR98 and ATXN7. Based on pedigree analysis, 20 patients were autosomal recessive retinitis pigmentosa, 3 patients were autosomal dominant retinitis pigmentosa and 1 patient was X linked retinitis pigmentosa. 3/7 patients with USH2A mutations were identified as Usher syndrome. ConclusionsUSHZA is the main disease-causing of sRP patients in Ningxia region. 83.3% of sRP in this cohort are autosomal recessive retinitis pigmentosa.

Novel mutations in the USH2A gene in a family affected with Usher syndrome type 2

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.

THE CHARACTERISTICS OF Rb GENE MUTATIONS IN 108 CASES OF RETINOBLASTOMA

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）

Trends in hospitalization expenses of PCI surgery for coronary heart disease patient based on double breakpoint interrupt time series analysis

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.

Clinical features and identification of gene mutation in families with familial exudative vitreoretinopathy

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.

Application of interrupted time series analysis in hospital scientific research incentive

The interrupted time series analysis was used to evaluate the incentive effect of the management methods of the SCI thesis fund for scientific research in West China Hospital of Sichuan University. We found an increase in number of the SCI papers and the growth rate after the adoption of scientific research incentive measures, indicating that the management methods of the SCI thesis fund had the incentive effect of scientific research. The interrupted time series analysis could be used in the incentive analysis of scientific research.

Primary mutation detection of mitochondrial DNA(mtDNA)in Leber′s hereditary optic neuropathy patients

Optic atrophy,hereditary／diagnosis; Polymerase chain reaction; DNA,mitochondrial; Point　mutation; Sequence analysis

Genomic sequence analysis of severe acute respiratory syndrome coronavirus 2 and search for possible medicines

The severe acute respiratory syndrome coronavirus 2 is characterized by a long incubation period, strong infectivity and general susceptibility to the population. At present, there are no specific medicines that can treat coronavirus disease 2019. In order to increase the understanding of the molecular biology of severe acute respiratory syndrome coronavirus 2 and try to find effective treatments, we used SnapGene Viewer to analyze the genomic sequences of five strains of severe acute respiratory syndrome coronavirus 2 that published by National Genomics Data Center. The results showed that the genome length of this virus was about 29.8 kb and twelve open reading frames were predicted, and five nucleotide change sites were found in the open reading frames. In addition, we analyzed drugs used during the outbreak of severe acute respiratory syndrome, current drugs for the treatment of coronavirus disease 2019 and other possible drugs, to find some possible medicines with clinical treatment effects.