ABSTRACT
OBJECTIVE@#To investigate accuracy of the currently used strategies for intraocular pressure measurements for reflecting actual 24-hour intraocular pressure fluctuations.@*METHODS@#From September, 2018 to January, 2019, the patients with a suspected diagnosis of primary open angle glaucoma at our hospital were prospectively enrolled to receive 24-hour intraocular pressure monitoring using a Goldmann tonometer. With the intraocular pressure measurements at 0:00, 2:00, 5:00, 7:00, 8:00, 10:00, 11:00, 14:00, 16:00, 18:00, 20:00, and 22:00 as the gold standard (strategy 1), we compared the measurements taken at 5:00, 7:00, 10:00, 14:00, 18:00, and 22:00 (strategy 2) and at 8:00, 11:00, 14:00, and 16:00 (strategy 3) for their accuracy in reflecting 24-h intraocular pressure fluctuations.@*RESULTS@#A total of 41 patients (82 eyes) were enrolled in this study. The peak intraocular pressures measured using the 3 strategies were 21.09±4.15 mmHg, 20.54±4.10 mmHg, and 19.91±4.38 mmHg, respectively, showing significant differences among them (@*CONCLUSIONS@#For suspected cases of glaucoma, intraocular pressure measurements at 4 and 6 time points of a day can not precisely reflect the actual range of intraocular pressure fluctuations, and may lead to a missed diagnosis of glaucoma.
Subject(s)
Humans , Glaucoma/diagnosis , Glaucoma, Open-Angle , Intraocular Pressure , Prospective Studies , Tonometry, OcularABSTRACT
OBJECTIVE@#To analyze the clinical features and genetic mutations in a patient with mucolipidosis type II α/β by using next generation sequencing.@*METHODS@#Clinical data of the patient was collected. Genomic DNA of the patient and her parents was extracted by a standard method. The patient was subjected to targeted sequencing using an Ion Ampliseq panel, which included genes related to mucolipidosis and mucopolysaccharidosis. Suspected mutations were verified by Sanger sequencing.@*RESULTS@#Compound heterozygous mutations, namely c.1284+1G>T and c.1090C>T (p.Arg364*), were detected in the patient, which were respectively inherited from her mother and father. No other disease-causing mutation was detected in the patient. GNPTAB c.1090C>T was known to be pathogenic, while GNPTAB c.1284+1G>T is a novel mutation. The same mutations were not detected among 50 healthy controls.@*CONCLUSION@#The compound heterozygous mutations c.1284+1G>T and c.1090C>T (p.Arg364*) of GNPTAB gene probably account for the mucolipidosis type II α/β in the patient. NGS has a great value for the molecular diagnosis and typing of mucolipidosis.
Subject(s)
Female , Humans , High-Throughput Nucleotide Sequencing , Mucolipidoses , Genetics , Mutation , Transferases (Other Substituted Phosphate Groups) , GeneticsABSTRACT
Objective To observe the surgical effects of scleral buckling and vitrectomy for familial exudative vitreoretinopathy (FEVR).Methods 34 eyes of 27 patients with FEVR who underwent either scleral buckling or vitrectomy were enrolled in this study.There are stage 2B in 2 eyes (5.88％),stage 3B in 7 eyes (20.59％),stage4Ain 1 eye (2.94％),stage4Bin 16 eyes (47.06％),stage 5 in8 eyes (23.53％).5 eyes associated with rhegmatogenous retinal detachment.The surgical approaches had been chosen according to the disease stage,severity,extent and morphology of the proliferative membrane.13 eyes (stage 2B in 2 eyes,3B in 4 eyes,and 4 in 7 eyes) underwent scleral buckling and 21 eyes (stage 3B in 3 eyes,4 in 10eyes,and 5 in 8 eyes) underwent vitreoretinal surgery.The main outcome measurement was the anatomic status of the macula,which was recorded as attached,partially attached or remain detached.The mean follow up was (18.00 ± 14.61) months (range 4 to 60 months).Results Among 13 eyes received scleral buckling,the macula was attached in 2 eyes with stage 2B (15.38％),partially attached in 11 eyes (84.62％) including 4 eyes with stage 3B,1 eye with stage 4A and 6 eyes with stage 4B.Among 21 eyes received vitrectomy,the macula was attached in 8 eyes (38.10％) including 2 eyes with stage 3B,4 eyes with stage 4 and 2 eyes with stage 5;the macula was partially attached in 9 eyes (42.86 ％) inducing 4 eyes with stage 4 and 5 eyes with stage 5;the macula remained detached in 4 eyes (19.05％) including 1 eye with stage 3B,2 eyes with stage 4 and 1 eye with stage 5.Conclusion If the surgical approaches were chosen based on the stage of FEVR and the severity,extent and morphology of the proliferative membrane,the surgery is effective and beneficial to FEVR patients.
ABSTRACT
Objective To construct human surfactant protein B (SP-B) gene promoter luciferase reporter plasmids and detect their transcriptional activities in H441 cells.Methods (1)The fragment of SP-B promoter (-218/+ 435 bp) was acquired from human genome DNA by polymerase chain reaction (PCR) amplification and then was inserted into pGM-T vector by the T4 DNA ligase.The vector was transfected into TOP10 E.coli.The positive clone was identified by DNA sequencing.The identified target SP-B promoter sequence was cloned into pGL3-basic vector to construct the recombinant vector pGL3-basic-SP-B-promoter and was identified by enzyme digestion and sequencing; (2)The pGL3-basic-SP-B-promoter vector was converted into pGL4.17-SP-B-promoter vector through enzyme digestion.The identified recombinant vectors and control plasmid pRL-TK were transfected into H441 cells by lipofectamine 2000,and luciferase assays was performed using the dual-luciferase reporter assay system.Results The sequences of SP-B promoter in the recombinant luciferase reporter plasmids were consistent with the one published on Genebank.The firefly/renilla luciferase activity ratio of pGL3-basic/pGL4.17-SP-B-promoter vector (2.8 ± 1.1,66.5±3.8) was significantly higher than pGL3-Basic,pGL4.17 control vector (0.2 ±0.1,4.3 ±0.4) with statistical significance (t =4.182,27.419,P =0.000),respectively.The SP-B promoter activity of pGL4.17-SP-B-promoter vector was significantly higher than pGL3-basic-SP-B-promoter vector (t =27.712,P =0.000).Conclusions The pGL3-basic/pGL4.17-SP-B-promoter vectors are successfully constructed with SP-B promoter activity in H441 cells and pGL4.17-SP-B-promoter vector is the better choice for further study with higher luciferase activity.
ABSTRACT
Objective To construct two kinds of eukaryotic ccll expression vcctors pIRES2-EGFP-SP-B-C/T 1580 and evaluate their expressions in 293T cells,for the further study of relationship between polymorphism of surfactant protein B (SP-B) gene and bronchopulmonary dysplasia (BPD).Methods The eukaryotic pIRES2-EGFP-SP-B-C/T 1580 expression vectors were constructed by gene recombination,and identified by gene sequencing.The recombinant expression vectors were transfected into 293T cells by lipofectamine2000.The expression of green fluorescence protein in 293T cells was observed by fluorescence microscopy.The mRNAs and proteins of SP-B-C/T 1580 were tested and identified by reverse transcriptionpolymerase chain reaction-restriction fragment length polymorphism(RT-PCR-RFLP) and western blot.Results Two recombinant plasmids contained the complete cDNA of SP-B with the same sequence as in gene bank.The base of SP-B 1580 gene of pIRES2-EGFP-SP-B-C 1580 was C,that of pIRES2-EGFP-SP-B-T 1580 was T.After being transfected into 293T cells,highly efficient expression of SP-B-C/T 1580 gene was detected at mRNA and protein levels.Conclusions The pIRES2-EGFP-SP-B-C/T 1580 eukaryotic cell expression vectors were successfully constructed.