Background Selective laser trabeculoplasty (SLT) plays lowing-intraocular pressure (IOP)effect by irradiating pigmented trabecular cells selectively using 532 nm Nd:YAG laser.However,its affect pattern is not fully known.Objective This study was to establish a human trabecular cells (HTCs) model of SLT effect,and to observe the morphological changes of HTCs after they were exposed to different energies of SLT.Methods Immortalized HTCs were incubated in DMEM/F12 with pigmental granules for 16 hours (overnight) to prepare pigmented trabecular cells.The cells were irradiated with modified laser lens with the spot of 400 μm and emitting duration of 3 ns.The energy scale of 0.2 mJ was set for standardized energy and 0.1 mJ was used as the low energy.The morphology and ultrastructure of the cells were examined under the light microscope and transmission electron microscope 1 hour,4,8,12,24 hours after irradiation.Trypan blue staining and TUNEL fluorescence staining were used to evaluate the death and apoptosis of the cells after laser irradiation.Results The brown pigment particles were seen in cytoplasm around nuclei of trabecular cells after cultured for 16 hours.After laser irradiation,there was no obvious change in the shape of nonpigmented trabcular cells,but a 400 μm spot was found in the pigmented trabecular cells under the light microscope,and the pigmented particles released as the cell disruption.After 0.2 mJ laser irradiation,cell loss zone was seen at the center of the laser spot,and the positive cells for trypan blue and TUNEL staining were found;while the abnormal manifestations were slight after the 0.1 mJ irradiation.With the lapse of the time,the number of positive staining cells was gradually decreased.Twenty-four hours after laser irradiation,proliferative trabecular cells emerged and migrated to the center area of laser spot.Statistical analysis showed that the numbers of the positive cells for trypan blue and TUNEL staining were significantly reduced in the lower energy group in comparison with the standardized energy group at various time points (all at P＜0.001),and those of both energy groups were gradually reduced with lapse of time with the significant differences between any two time points (all at P＜0.01).Conclusions An in vitro laser effect model of HTCs can be established by exposing pigmented HTCs to SLT laser.After exposed to SLT,the gasification,necrosis-like death,apoptosis-like change appear at the laser center.The laser destroyed cells can be replaced by proliferative cells with the lapse of time.Low-energy laser irradiation cause a similar morphological change of the cells to high-energy irradiation,but the number of abnormal cells is much less.

Background Selective laser trabeculoplasty (SLT) can increase the outflow of aqueous humor and reduce the intraocular pressure of patients with open angle glaucoma,but its mechanism is unknown.To investigate the mechanism of SLT would improve the therapeutic effect of SLT.The aqueous outflow resistance in trabecular meshwork was affected by the extracellular matrix (ECM).Matrix metalloproteinase-3 (MMP-3) and MMP-9 were closely related to ECM degradation in trabecular meshwork.Objective This study was to observe the effects of SLT on the expressions MMP-3 and-9 in human trabecular ceils in vitro.Methods Immortallized human trabecular cells were cultured with pigment particles mixed suspension for 16 hours and incubated overnight.Then the cells were irradiated with Q switch frequency doubling 532 nm Nd:YAG laser to establish SLT-effective cells with the energy of 0.2 mJ,spot diameter of 400 μm and pulse duration of 3 ns.The expressions of MMP-3 mRNA and MMP-9 mRNA in the cells were detected by fluorescence quantitative real time PCR before and 1 hour and 4,8,12,28 hours after exposure of laser.The concentrations of MMP-3 and MMP-9 in the medium were assayed using ELISA 1 hour and 4,8,12,28 hours after exposure of laser and compared between the non-irradiation group and the irradiation group.Results The relative expressing levels of MMP-3 mRNA were 1.00,1.32±0.12,2.08±0.05,2.34±0.04,2.77± 0.05 and 2.49±0.27 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of laser SLT,showing a significant difference among the groups (F =15.966,P=0.007),and relative expressing levels of MMP-3 mRNA were significantly higher in various time points after laser irradiation than those of the non-irradiation group (all at P＜0.05).The relative expressing levels of MMP-9 mRNA were 1.00,0.91 ±0.10,1.27 ± 0.07,1.46±0.07,1.69±0.09 and 0.87±0.09 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of SLT,which was considerably different among the groups (F =30.526,P =0.005),and significant increased values were seen in the 4,8 and 12 hours after irradiation compared with the non-irradiation group (all at P＜0.05),with highest expression in the irradiation for 12-hour group.The concentrations of MMP-3 and MMP-9 proteins in medium were significantly increased in various time points after laser exposure in comparison with the control group (all at P＜0.05).Conclusions The expressions of MMP-3 and MMP-9 in human trabercuolar cells upregulate and the secretion ability of human trabecular cells to MMP-3 and MMP-9 proteins improves in early stage of SLT in vitro.However,these procedures gradually diminish with the lapse of time.

BACKGROUND: Therapeutic effect on pterygium mainly focuses on studying surgical technique, assistant therapy methods,and recurrence rate following excision of pterygium; however, whether race factor is associated with occurrence, development, and recurrence of pterygium remains still unknown.OBJECTIVE: To compare the therapeutic outcomes in different race patients with pterygium treated using corneal limbal stem cell autograft combining with excision of pterygium.DESIGN, TIME AND SETTING: Retrospective case analysis, performed in the Department of Ophthalmology, Zhongshan Hospital Affiliated to Xiamen University between January 2000 and June 2006. PARTICIPANTS: 1 44 (152 eyes) primary cases were collected from Xiamen and 54 (54 eyes) relapsed Negroes were from Africa. There were no significant differences in age, sex, and pterygium length between the two groups (P 0.05).METHODS: 198 subjects were treated by excision of pterygium under a microscope, in which pterygium tissue was not found on the surface of cornea. A free transplantation of the superotemporal limbus with an adjacent piece of thin conjunctiva was placed in the excision area. All cases were followed-up to grade the appearances of the sites 6 weeks, 6 and 12 months after excision (grade 1 implied normal appearance, and grade 4 implied the relapse). MAIN OUTCOME MEASURES: Relapse, pterygium grading, and complication after corneal limbal stem cell autograft combining with excision of pterygium. RESULTS: No relapse was found at 6 weeks after operation. The recurrence rate of Chinese Han people and Africa black people were 6.6% and 14.8% respectively at 6 months, and 11.8% and 24.5% at 12 months. There was significant difference in the recurrence rate between the two races (t=4.607, P= 0.032). In addition, there were significant differences in the pterygium grading between the two groups at 6 weeks (x2=15.608, P < 0.01 ), and Chinese people recovered better. Contrarily, there was no statistical difference at 6 months and 12 months (x2=4.401, 6.206; P 0.05). Few complications were found except superficial scar of cornea and persistent irritation of ocular surface. CONCLUSION: Limbal stem cell autograft combining with excision of pterygium under a microscope can completely remove pterygium with minimal invasion, light postoperative response and low rate of recurrence. The relapse of black cases is higher than Chinese patients.

Ataxins ; Humans ; Male ; Middle Aged ; Nerve Tissue Proteins ; genetics ; Spinocerebellar Ataxias ; genetics ; Trinucleotide Repeats

Objective To identify the specific genotype and analyze clinical features of spinocerebellar ataxias (SCAs) pedigree in the region of Yunnan Province.Methods Fourteen SCAs pedigrees and 183 blood samples of the family members were collected between January 2011 and July 2014 from Department of Neurology,First Affiliated Hospital of Kunming Medical College.Polymerase chain reaction (PCR) amplification,agarose gel electrophoresis and DNA sequencing technologies were utilized to identify the specific genotype of SCAs pedigree.Presymptomatic tests were carried out and the clinical features and genetic test results of patients were carefully analyzed.Results SCA3 was the most common subtype of SCAs in the Han nationality of Yunnan region.Nine of the 14 families were SCA3,only one family was SCA2.Additionally,there were four SCAs families that remained indeterminate.The patients with di-allele mutations (46/77) of SCA3 gene had early onset,rapid progression and serious clinical symptoms.Hereditary SCA3 and autonomic dominant polycystic kidney disease can happen simultaneously in a family.The proband SCA3 gene' s CAG repeat number is 28/76,and repetitions of the mutation allele are in all range.The PKD1 gene exon 23 is found to be in abnormal sequence.Conclusions SCA3 is the most common subtype of SCAs in the Han population of Yunnan region.There are 15/46 incomplete penetrance nutation and 46/77 di-allele mutations.It is possible that di-allele mutations make the disease worse and accelerate clinical course progression.SCA3 and polycystic kidney disease can uncommonly happen simultaneously in a family,which perhaps suggests there are interactions between the two disease-virulence genes.

Purpose To explore the predictive value of myocardial perfusion in assessing myocardial systolic function recovery after primary percutaneous coronary intervention (PPCI),in order to improve poor prognosis by early detection of myocardial no-reflow.Materials and Methods Forty nine patients with acute myocardial infarction (AMI) who had received PPCI were chosen as subjects.All the patients underwent two-dimensional strain (2DS) images and resting real-time myocardial contrast echocardiography (MCE) within one week after surgery,and 2DS measurement was repeated after three months.2DS imaging was used to acquire longitudinal peak systolic strain (LPSS) at all myocardial segments.Based on the graphs of LPSS,left ventricular myocardium was divided into normal contractile function myocardium (red) and impaired contractile function myocardium (light red,blue).According to the myocardial perfusion scores (MPS) qualitatively assessed by MCE visual interpretation,the myocardia with impaired systolic function were categorized into three groups with different perfusion level.The changes of LPSS within one week and three months after surgery (△ LPSS) among the three groups were analyzed.The correlation between MPS and LPSS within one week and three months after PPCI was also analyzed respectively.Results The △ LPSS increased significantly among the three groups with the improvement of myocardial perfusion level [(-5.78±6.23)％ vs.(-4.37±6.60)％ vs.(-1.21 ±4.77)％,all P＜0.05].The MPS measured one week after PPCI was both positively correlated with the LPSS detected within one week after surgery and that after three months (r=0.47,0.58,P＜0.001).The consistence of myocardial perfusion scores given by two evaluators was good (Kappa=0.785,P＜0.05).Conclusion The level of myocardial perfusion after PPCI in patients with AMI is closely related to regional myocardial systolic function,and the improvement of myocardial perfusion can forecast the recovery of regional systolic function.

Objective To explore the distribution characteristics of ankle-brachial index ( ABI) examination for peripheral arterial dis-ease in diabetic patients, and to provide basis for clinical diagnosis. Methods Randomly selected 110 cases of patients who were admitted into our hospital from February 2014 to January 2015, and divided them into the observation group (34 cases with diabetic lower extremity peripheral vascular lesions) and the control group (76 cases without diabetic lower extremity peripheral vascular lesions). Measured the ABI of the two groups, and analyzed the value of ABI data range for the diagnosis of diabetic lower extremity peripheral vascular lesions. Mean-while, analyzed the risk factors of peripheral arterial disease in diabetic patients according to ABI. Results The sensitivity of ABI for diag-nosis of diabetic lower extremity peripheral vascular lesions was 91. 18%, and the coincidence rate was 95. 36%, the specificity was 98. 68%, the misdiagnosis rate was 8. 82%, and the omission diagnostic rate was 8. 82%. ABI of the observation group was obviously lower than that of the control group, and the difference was statistically significant (P<0. 05). Conclusion ABI examination could be basis of clinical diagnosis and important standard of disease assessment for peripheral arterial disease in diabetic patients. It is suggested to strengthen the control of blood glucose, blood pressure and weight of patients with diabetes in order to reduce the risk of morbidity.

Objective To evaluated the value of myocardial perfusion before delayed percutaneous coronary intervention (PCI) for predicting the recovery of systolic function of patients with acute myocardial infarction (AMI).Methods A total of 64 patients with AMI receiving delayed PCI treatment in the First People's Hospital of Foshan from January 2014 to June 2015 were selected.One day prior to delayed PCI,all of the patients underwent two dimensional strain to measure the longitudinal peak systolic strain (LPSS) of each left ventricular segment and the global longitudinal strain (GLS) of the left ventricle.The myocardial perfusion score (MPS) and the perfusion score index (PSI) were measured by myocardial contrast echocardiography (MCE).Left ventricular myocardial perfusions were classified as good,reduced,or absent.The two dimensional strain measurements were again conducted at 6 months after the delayed PCI to assess LPSS and GLS.The change of GLS and LPSS between one day prior to delayed PCI and six months after delayed PCI was assessed by paired t-test.The differences of LPSS among good,reduced,or absent myocardial perfusion groups were analyzed by one-way ANOVA.LSD-t test was used to compare in pairs of groups that had different values.The correlations between PSI and GLS,MPS and LPSS were assessed by Spearman's rank-correlation test.Results The GLS of all patients were higher at six months after delayed PCI than at one day prior to delayed PCI [(-15.39±7.80)％ vs (-12.44±8.38)％,t=14.398,P ＜ 0.001].The LPSS of myocardial perfusion in good,reduced and absent groups at one day prior to delayed PCI were (-2.64±5.60)％,(-6.19±6.87)％ and (-12.07±5.86)％,respectively.The LPSS of myocardial perfusion in good,reduced and absent groups at six months after delayed PCI were (-2.97 ± 4.93)％,(-11.38± 7.26)％ and (-15.82 ± 5.97)％,respectively.The myocardial LPSS of left ventricular segment with good or reduced perfusion was significantly higher at six months after delayed PCI (t=13.013,10.821,both P ＜ 0.001),but the LPSS of left ventricular segment with absent perfusion was similar to that of pre-PCI.Whether at one day prior to delayed PCI or six months after delayed PCI,there were significant differences in LPSS parameters among the three groups (at one day prior to delayed PCI,myocardial perfusion absent vs reduced or good,t=4.201 and 11.771,both P ＜ 0.001;myocardial perfusion reduced vs good,t=12.561,P ＜ 0.001;at six months after delayed PCI,myocardial perfusion absent vs reduced or good,t=9.714 and 15.646,both P ＜ 0.001;myocardial perfusion reduced vs good,t=9.254,P ＜ 0.001).The LPSS both at one day prior to delayed PCI and six months after delayed PCI in myocardial perfusion good group ＞ those of myocardial perfusion reduced group ＞ those of myocardial perfusion absent group.PSI was positively correlated with GLS at both one day prior to delayed PCI and six months after delayed PCI (r=0.69,0.72,both P ＜ 0.001).MPS was positively correlated with LPSS at both one day prior to delayed PCI and six months after delayed PCI (r=0.49 and 0.45,both P ＜ 0.001).Conclusion Myocardial perfusion before delayed PCI,monitored by MCE,is correlated well with myocardial systolic function,and may be used to predict the recovery of myocardial systolic function after delayed PCI.

Multiple sequence alignment is one of the basic techniques in bioinformatics, and it plays a vital role in structure modeling, functional site prediction, and phylogenetic analysis. In this paper, we review the methodologies and recent advances in the multiple protein sequence alignment, e.g., speeding up the calculation of distances among sequences and employing the iterative refinement and consistency-based scoring function, with emphasis on the use of additional sequence and structural information for improving alignment quality.
Algorithms ; Proteins ; chemistry ; Sequence Alignment ; methods ; Sequence Analysis, Protein ; methods

OBJECTIVETo investigate clinical features and genetic mutations of a family affected with spinocerebellar ataxia 3 and polycystic kidney disease.

METHODSPolymerase chain reaction and DNA sequencing were employed to analyze exon 10 of the SCA3 gene, in addition with all exons and flanking sequences of PKD1 and PKD2 genes. The clinical features were also carefully analyzed.

RESULTSThe numbers of CAG repeat in the proband's SCA3 gene were 28/76, with the number of repeats in the mutant allele being in the full range. The sequence of exon 23 of the PKD1 gene was also found to be abnormal. Clinical symptoms of the proband were very serious, which were characterized by obvious ataxia, pyramidal signs, Meige syndrome, depression and high blood pressure.

CONCLUSIONHereditary spinocerebellar ataxia 3 and autonomic dominant polycystic kidney disease may co-occur, and genetic testing is the primary means of diagnosis.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Polycystic Kidney Diseases ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spinocerebellar Ataxias ; genetics