PURPOSE:To investigate mitochondrial DNA(mtDNA) of Leber's hereditary optic neuropathy(LHON). METHODS:Polymerase chain reaction(PCR)method was used to analyse mtDNA of 11 patients in a pedigree with LHON and 4 control subjects from none LHON pedigree. RESULTS:There was a loss of a restriction site for the restriction endonuclease SfaN.Ⅰin Ihe Patients with LHON. In this pedigree,maternal lineage was regarded a carrier of the pathogenic gene. CONCLUSIONS:The patients with Leber's hereditary optic neuropathy have a point mutation in mtDNA,which results in loss ol SfaN I endonuclease restriction site .and this change is one of mechanisms inducing this disaese. （Chin J Ocul Fundus Dis,1997,13: 27-29）
Objective To investigate the mutations of the gene in Chinese patients with X linked juvenile retinoschisis (XLRS), and to provide the genetic diagnosis and consultation of heredity for the patients and their families. Methods Genomic DNA was isolated from leukocytes of 29 male patients with XLRS, 38 female carriers and 100 normal controls (the patients and the carriers were from 12 families). All 6 exons of XLRS1 gene were amplified by polhism (SSCP) assay. The positions and types of XLRS1 gene mutations were determined by direct sequencing. Results Eleven different XLRS1 mutations were identified in these 12 families, including one frameshift mutation due to base loss of the first exon: c.22delT（L9CfsX20）, one nonsense mutation due to base loss of the first exon (Trp163X), one splice donor site mutation(c.52+2 Trarr;C; IVS1+2T to C), and eight missense mutation due to base replacement（Ser73Pro, Arg102Gln, Asp145His, Arg156Gly, Arg200Cys, Arg209His, Arg213Gln, and Cys223Arg). No gene mutation was detected in the control group. Four new mutations included frmaeshift mutation（L9CfsX20）and mutations of Asp145His, Arg156Gly, and Trp163X at the fifth exon. A newly discovered non-disease-related polymorphism (NSP) was the c.576C to T (Pro192Pro) change at the sixth exon. Conclusion Eleven different XLRS1 mutations were detected, which is the cause of XLRS in Chinese people. The detection of gene mutations may provide the guidance of genetic diagnosis and the consultation of family heredity for the patients and their families. (Chin J Ocul Fundus Dis, 2006, 22: 77-81)
Objectives To study the relationships between methylenetetrahydrofolate reductase(MTHFR ) gene polymorphism and diabetic retinopathy (DR) in Chinese Han race． Methods With polymerase chain reaction and restriction fragment length polymorphism (PCR-FLP), MTHFR gene 677 T mutation (cytosine is replaced by thymine in No. 677 site) was detected in 85 health controls, 62 with DR and 117 without DR of type 2 diabetics comfrimed by ophthalmoscope． Results The frequency of MTHFR variant genotypes and alleles of DR in Chinese Han race．patients were signigicantly higher than those without retinopathy and healthy controls (Plt;0．01)． Conclusions The results suggested that MTHFRgene C677T mutation was probably one of the genetic risk factors of diabetic retinopathy in Chinese Han rase． (Chin J Ocul Fundus Dis, 2001,17:198-200
Objective To evaluate the potential of specific mRNA marker keratin 19(K19) to detect micrometastasis by reverse transcriptase polymerase chain reaction (RT-PCR) .Methods One hundred and ninty four regional lymph nodes harvested from 6 cases of benign diseases, 4 cases of breast carcinoma, 5 cases of gastric carcinoma and 12 cases of colorectal carcinoma patients were examined by conventional pathology and amplifying tissue specific K19 mRNA by RT-PCR separately, then the two methods were compared with each other. Results None of the 34 lymph nodes which were pathological metastasis-negative from benign diseases expressed K19 mRNA by RT-PCR, all of the 28 regional lymph nodes which were pathological metastasis-positive from malignant cases showed trains of K19 mRNA by RT-PCR. Of the 132 lymph nodes which were pathological metastasis-negative from malignant cases, 11 lymph nodes were detected with micrometastasis by genetic diagnosis.Conclusion Genetic diagnosis of lymph node micrometastasis is more sensitive than conventional pathology and has diagnostic value and merits further study.
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)
To investigate the mRNA expression of nm23-H1 gene in human liver tumor. In tumor and corresponding nontumoral liver specimens from 20 patients, nm23-H1 mRNA were examined by reverse transcriptionpolymerase chain reaction (RT-PCR) method with specific primers. Results: The primers designed in this study could amplified nearly entire coding sequence of nm23-H1 gene. All the samples showed positive expression of nm23-H1 mRNA, indicating there was no expression loss or obvious alteration. Conclusions: The achievement of RT-PCR method lays foundation for quantitative gaugement of nm23-H1 mRNA in liver tumor.
Objective To review the appl ication of collagen and biodegradable polymer composite scaffolds in vascular tissue engineering, and describe the multi-layering vascular scaffolds of collagen-based material in recent years. Methods The l iterature concerning collagen composite scaffold production for scaffold of vascular tissue engineering was extensively reviewed and summarized. Results As one of the structural proteins in natural blood vessel, collagen is widely used in vascular tissue engineering because of good biocompatibil ity, biodegradabil ity, and cell recognition signal. The vascular scaffolds with biological activity and good mechanical properties can be made by collagen-polymer composite materials. In addition, the structure and function of the natural blood vessel can be better simulated by multi-layering vascularscaffolds. Conclusion Collagen-polymer composite material is the hot spot in the research of vascular scaffolds, and multilayering vascular scaffolds have a brill iant future.
Objective To construct specifically expressed vascular endothelial growth factor (VEGF)165 gene in retina. Methods Rho promoter, specifically expressed in retina, was amplified by polymerase chain reaction (PCR) from the genomic DNA of a BLAB/C rat, then it was cut with restriction enzymes and cloned into the plasmid pcDNA3.1+-VEGF165 to form recombinant plasmid pcDNA3.1+-rho-VEGF165. The correct recombinant plasmid pcDNA3.1+-rho-VEGF165 was identified by restriction enzymes and PCR, and was transferred by jetPEI into cultured human navel vein endothelial cells and human retinal pigment epithelial (RPE) cells. The expression of VEGF protein in human navel vein endothelial and RPE cells was detected by immunocytochemical staining and protraction of the growth curve of the cells. Results In human RPE cells, the expression of VEGF protein was more in recombinant plasmidpcDNA3.1+-rho-VEGF165 than that in plasmidpcDNA3.1+-rho-VEGF165 ; in human navel vein endothelial cells, no obvious difference of the expression of VEGF protein between recombinant plasmid pcDNA3.1+-rho-VEGF165 and plasmid pcDNA3.1+-rho-VEGF165 was found. Conclusions The construction of pcDNA3.1+-rho-VEGF165 carrier may provide the basic material for the study of the nosogenesis of VEGF in retinal neovascularization, and establish the foundation to set up the model of transgenic mice with VEGF specific expressing in retina. (Chin J Ocul Fundus Dis, 2005,21:106-108)
OBJECTIVE: To investigate the availability of bone defect repair by degradable porous polycaprolactam (PCL) as the carrier of bone morphogenetic protein (BMP). METHODS: Three different kinds of bone substitutes, including decalcified bone matrix, PCL-BMP compounds and simple PCL, were implanted into the radial bone defects in 36 rabbits, and in the other 12 rabbits the bone defects were left untreated as control. X-ray examination, X-ray morphometry, histological and electron microscopic observation were performed at different time after operation. RESULTS: The quantity of new bone formation in PCL-BMP group was prior to that in simple PCL group. The pattern and speed of bone defect repair in PCL-BMP group were similar to those in decalcified bone matrix. Electron microscopic observation showed that the PCL-BMP group degraded faster than simple PCL group, which was more suitable for bone repair. CONCLUSION: PCL is a good carrier of BMP with potential for clinical use.
Objective To analyze the association of human leucocyte antigen (HLA)-DRB and -DQB alleles with Ealesprime; disease, and to investigate the potential immunogenetics mechanism of Ealesprime; disease. Methods Gene loci of HLA-DRB and -DQB1 alleles were detected by polymerase chain reaction-sequence specific primer (PCR-SSP) in 27 Han-nationality patients with Ealesprime; disease in Northern China and 30 age and sex-matched normal persons as control, then statistics package for social science (SPSS) for Windows ver 13.0 software was used to analyze the distribution features of frequency of HLA-DRB and -DQB1 alleles in the two groups. Results Compared with the control group, the frequency of HLA-DRB104 allele increased obviously in the patients with Ealesprime; disease［odds ratio (OR)=3.20 ，OR 95% confidence interval（CI）=1.00-10.21, and P=0.047］. Simultaneously, no statistically significant difference of the distribution of any other DRB or DQB1 allele between the two groups was found (Pgt;0.05). Conclusions In hannationality people in Northern China, DRB104 is positively associated with Ealesprime; disease, suggesting that DRB104 may confer a major influence on Ealesprime; disease. Turbulence of immune function begotten by infect-agents attack may occur in the individuals with Ealesprime; disease due to the specific hereditary diathesis of HLA, which may cause the occurrence and development of Eales disease. (Chin J Ocul Fundus Dis, 2006, 22: 90-93)