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 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）
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.
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.
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)
Objective To explore the impact of diagnosis-related group (DRG) payment method reform under total amount control on neurology and neurosurgery departments. Methods The DRG grouping data of the Department of Neurology and the Department of Neurosurgery of Panzhihua Central Hospital from January 2018 to December 2020 were collected, and the mature DRG evaluation indexes in China were selected. Using the interrupt time series analysis method, the DRG-related indexes of the two departments before and after the introduction of the performance appraisal plan in July 2019 were compared, to evaluate the intervention effects on the two departments. Results Both neurology and neurosurgery departments showed a slow downward trend in the overall medical service capacity under the DRG payment. The efficiency of medical services showed a slow upward trend and the consumption of medical expenses showed a slow downward trend in the Department of Neurology, while the efficiency of medical services showed a slow downward trend and the consumption of medical expenses showed a slow upward trend in the Department of Neurosurgery. According to the results of interrupt time series analysis, in the Department of Neurosurgery, the total weight showed a significant downward trend before intervention (β1=−5.526, P=0.003), and the downward trend became sluggish after intervention, with a statistically significant slope difference before and after intervention (β3=4.546, P=0.047); the case-mix index showed a downward trend before intervention (β1=−0.050, P<0.001), and no obvious trend after intervention, with a statistically significant slope difference before and after intervention (β3=0.052, P=0.001); the cost consumption index showed no obvious downward trend before intervention (β1=−0.006, P=0.258), and an upward trend after intervention, with a statistically significant slope difference before and after intervention (β3=0.027, P=0.032). The impact of this assessment plan on the Department of Neurology was not statistically significant (P>0.05), needing further observation. Conclusions The reform of DRG payment method under total amount control has different effects on the evaluation indicators of clinical departments of different natures. It is recommended to implement classified management and assessment for clinical departments of different natures.
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 construct eukaryotic expression vector of pEGFP-N3-TFPI-2, and to provide the base of studying the function of TFPI-2 gene. MethodsExtraction of total RNA from placental tissue was extracted at first, and then reverse transcriptase synthesis of cDNA was carried out. The cDNA fragment of TFPI-2 gene which was obtained by real time PCR (RT-PCR) was inserted into eukaryotic expression vector of pEGFP-N3. After double digestion with XhoⅠand KpnⅠ, the recombinant vector of pEGFP-N3-TFPI-2 was identified in 1% agarose gel electrophoresis and was tested by the sequence analysis. Then, the recombinant vector of pEGFP-N3-TFPI-2 (transfection group) and vector of pEGFP-N3 (blank control group) were transfected into Top10 competent cells with LipofectamineTM 2000, but no transfection-related treatment was performed in cells of untransfection group. Western blot method was used to test the expression of TFPI-2 protein in cells of 3 groups. ResultsThe purity of total RNA which were analysis by agarose gel electrophoresis and spectrophotometry were fit for PCR. After coding of TFPI-2 gene fragment and eukaryotic expression vector of pEGFP-N3, the recombinant plasmid of pEGFP-N3-TFPI-2 were got double digestion with XhoⅠand KpnⅠ, and was identified in 1% agarose gel electrophoresis, of which showing that there were 2 specific amplification of strips at 708 bp and 4 700 bp. Result of sequence analysis confirmed that the size of recombinant vector was consistent with the theoretical value. Results of Western blot showed that the expression of TFPI-2 protein in transfection group (0.657 3±0.032 5) was higher than those of blank control group (0.301 7±0.028 7) and untransfection group (0.314 3±0.026 6), P < 0.01. ConclusionsThe eukaryotic expression vector of pEGFP-N3-TFPI-2 has been constructed successfully, which laiding the foundation for the analysis about function of TFPI-2 gene.
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.