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)
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)
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.
ObjectiveIn light of the comprehensively implemented reform of medical insurance payments, this study analyzed the impact of the payment intervention and COVID-19 pandemic on hospitalization expenses for identical diseases between traditional Chinese medicine (TCM) and Western medicine hospitals, to provide evidence to promote high-quality coordinated development of hospitals and insurance while reducing patient load. MethodsFrom January 2014 to December 2020, we gathered data including 9 900 individual medical records of woman-related malignant tumors (WMT) from all 23 public hospitals in a district of Shanghai. We developed an interrupted time-series analysis model based on the above two interventions, to compare the inpatient average per-time expenses between different hospitals and different groups. ResultsThe average per-time expenses of WMT in Western hospitals changed from rising to declining after the policy intervention, and increased again during the pandemic. In TCM hospitals, the expenses continued to increase and fluctuated after the pandemic. ConclusionThe policy intervention has achieved a good effect on controlling the cost of Western hospitals, rather than the significant increase in TCM hospitals. Meanwhile, the COVID-19 pandemic has had a significant impact on hospitalization expenses. It’s urgent to develop a payment model that fits the development and characteristics of TCM, to control the unreasonable growth of expenses. Moreover, the financial compensation methods and supervision mechanism of public hospitals should be improved to effectively resist the threat of public health emergencies for the development of hospitals and the legitimate rights of patients.
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）
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 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.
Optic atrophy,hereditary／diagnosis; Polymerase chain reaction; DNA,mitochondrial; Point mutation; Sequence 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.