ObjectiveTo explore the variation of the structure of the intestinal flora between healthy people and patients with obstructive jaundice perioperatively. MethodsFrom February 2013 to August 2014, 20 patients with obstructive jaundice and 10 healthy persons (normal control group) in our hospitol were selected as the research object. The first stool specimens of the research object after admission were obtained and the total fecal bacteria DNA were extracted. After polymerase chain reaction amplification, the changes in the structure of bacterial flora were dynamic observed by using denaturing gradient gel electrophoresis (DGGE), and the gel bands were analyzed by using Quantity One software. The similarity and diversity of flora structure, and principal component analysis (PCA) were analyzed. ResultsSignificant differences of colonic microflora were found between patients with obstructive jaundice and healthy people; advantage intestinal flora in obstructive jaundice patients was significant lower than the normal control group. With the extension of time and degree of obstruction aggravated, a descending trend was found in number, abundance, and diversity of the intestinal microflora (P < 0.05). ConclusionThere is significant differences in the structure of colon bacteria in patients with obstructive jaundice and healthy persons.
Objective To evaluate the rapid diagnosis of bacterial and (or) fungal endophthalmitis by multiplex polymerase chain reaction (MPCR). Methods MPCR was performed to detect the DNA segment of bacteria and (or) fungi from standard strains and 41 samples of intraocular fluid or vitreous from 38 patients (3 with double eyes and 35 with single), and the results were compared with the cultured bacteria and fungi. Results Five hours after detected by MPCR, bacteria and (or) fungi in 34 out of 41 samples (82.9%) from patients were detected，in cluding bacteria in 26，fungi in 6，and both bacteria and fungi in 2. The positive rate of MPCR was obviously higher than the cultured ones（χ2＝9.60, P＜0.05）. Conclusion With the advantages of rapidity, sensibility, and specificity, MPCR can make for the rapid and definitive diagnosis of bacterial and (or) fungal endophthalmitis. (Chin J Ocul Fundus Dis，2004，20：81-83)
Purpose To investigate the relationship between mitochondrial DNA 11778 mutation and clinical characteristics of patients with Laber is hereditary optic neuropathy(LHON). Methods PCR RFLPs (MaeⅢ) and mutation specific primer PCR(MSP-PCR) were used simultaneously to detect mitochondrial DNA 11778 mutation. Results Among 10 subjects who habored 11778 mutation,one was a carrier and nine were patients with LHON.Of the nine patients,six were males and three were females.The age of onset ranged from 12 to 25 years old and the onset interval of the two eyed varied between 0 to 6 months. The visual acuity was CF/10cm-0.1 except one who lost her vision after delivery but recovered gradually.The results of visual field,VEP and color vision were abnormal but ERG and systemic status were all normal. Conclusion Molecular biological detection of the ten subjects showed that they all habored mtDNA 11778 mutation.The existence of carrier and visual recovery imlied that mtDNA mutation was a primary cause of LHON,but other factors such as endocrine disorder might influence the pathogenesis of LHON. (Chin J Ocul Fundus Dis,1998,14:156-158)
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 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)
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)
Objective To probe the association between HLA-B27 subtype and acute anterior uveitis(AAU). Methods The HLA-B27 gene of 49 AAu patients were inspected by means of polymerase chain reaction(PCR), and the B27 subtypes of AAU patients were analyzed by means of DNA sequencing and simulated nucleotide hybridization in computer. Results Twenty-nine(59.39%)of 49 AAU patients were B27 positive, and among them 13 (44.00%)were B2704 and 16(56.00%)B27052 in subtypes．No statistically significant difference in most clinical features were found between the patients with B2704 and B27052.In B27 pasitive patients,8 (seven is B27052 and one is B2704) of them were complicatde with ankylosing spondylitis(AS). Conclusions The AAU patients with either B2704 or B27052 have similar clinical feature.The B27052 gene may be an important factor for complicated AS in AAU patients. (Chin J Ocul Fundus Dis, 1999, 15: 139-142)
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 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 To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)