1.Influence of monochromatic light exposure on the growth of Müller cell and the expression of myopia-related cell factors in vitro
Jing, XIA ; Xiao-feng, ZHANG ; Wei, XIA ; Lei, ZHONG ; Zheng-tai, SUN ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2013;(5):446-451
Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Müller cells participate in the formation of myopia.However,the effect and mechanism of retinal Müller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Müller cells and the expression of cell factors in Müller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Müller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Müller cells were cultured with DMEM containing 10% fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Müller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Müller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P>0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Müller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia.
2.Experimental study on the eyelid reconstruction in situ with the silk fibroin membrane
Lei, ZHONG ; Xiao-feng, ZHANG ; Zheng-tai, SUN ; Jing, XIA ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2011;29(11):968-972
Background Autologous and allograft renal transplantation exist some disadvantages of less donor source and rejection.As a scaffold of cell in tissue engineering,fibroin was determined to have a good biocompatibility.But whether the fibroin membrane can become a substitution for tissue defect is seldom reported.Objective This experiment aimed to investigate the feasibility of silk fibroin membrane in the rabbit eyelid reconstruction in situ.Methods A 4 mmx3 mm tarsi defect model was created on the upper eyelids of 18 healthy New Zealand white rabbits.The eyelid reconstruction in situ was performed with regenerated silk fibroin membrane material in the right upper eyelids (silk fibroin group ) and allogenic sclera material (sclera group ) on the upper eyelids of fellow eyes.The grafts were clinically examined for the evaluation of inflammation and implant exposure at the first,second and forth week after operation.The inflammation response and collagen distribution were examined by hematoxylin & eosin staining and Masson staining.Expression of basic fibroblast growth factor(bFGF) in the grafts was detected by immunohistochemistry,and ImagePro Plus software was used for statistical analysis.Results All eyelid defects showed a primary healing.The surface of palpebral conjunctival was smooth and the inflammation of ocular surface was mild.The eyelid margin in the sclera group was more notch than that in the silk fibroin group.Results of pathological examination revealed that the arrangement of collagen fibers in the sclera group was more disordered,but that in the silk fibroin group was regular.The expression level(A value) of b-FGF in the operative area in silk fibroin group were 0.027 67±0.004 69,0.051 73±0.008 72,0.058 72±0.006 88,and those in the sclera group were 0.056 48±0.009 14,0.072 83 ± 0.009 17 and 0.078 73 ±0.010 84 in 1,2,4 weeks after operation,showing statistically significant differences between two groups in various time points ( t =- 6.38,t =- 4.99,t =- 2.87,P <0.05 ).Conclusions Silk fibroin membrane can reconstruct the eyelid shape in situ with the less inflammation response and good biocompatibility.Silk fibroin membrane could be used to support the eyelid as a new tarsal repairing materials.
3.Effect of laser in situ keratomileusis with femtosecond laser on visual quality
Xiao-Hua, LEI ; Chang-Tai, YU ; Ying, ZHANG ; Jing, LI ; Miao, MA
International Eye Science 2016;16(6):1120-1123
?AIM:To compare the changes of the contrast sensitivity after LASIK with femtosecond laser and microkeratome and to explore the influence of different methods making corneal flap on visual quality.? METHODS: There were 212 eyes in 106 myopes underwent excimer operation . According to the different methods of operation, they were divided into two groups:microkeratome group ( SBK group ) and femtosecond laser group ( FS group) . FS group: a total of 112 eyes in 56 patients received LASIK with femtosecond laser. SBK group: a total of 100 eyes in 50 patients received LASIK with microkeratome. Contrast sensitivity was detected preoperatively, and 1wk, 3mo postoperatively and compared between the two groups.? RESULTS: At 1wk after operation, the contrast sensitivity under photopic environment decreased in the two groups, compared with those before operation ( P<0. 05). The differences of contrast sensitivity before and 3mo after operation were not significant (P>0. 05). No statistical significant difference was found in contrast sensitivity under photopic environment at 1wk, 3mo between the two groups ( P>0. 05 ). At 1wk after the operation, the contrast sensitivity under scotopic environment decreased in both groups compared with those before operation ( P< 0. 05 ). In SBK group, it decreased more than in FS group (P<0. 05). After 3mo, the decline of 14. 2c/d spatial frequency contrast sensitivity under scotopic environment in the SBK group was more than other frequency. No statistical significant difference was found in the rest frequency contrast sensitivity under scotopic environment before and after operation (P>0. 05). After 1wk, contrast sensitivity with glare stimulation in both groups decreased, compared with those before operation ( P< 0. 05 ), while in SBK group, it decreased more than in FS group (P<0. 05). After 3mo, except that the decline of 14. 2c/d spatial frequency contrast sensitivity with glare stimulation in the SBK group was significant compared with those before operation, the contrast sensitivity under glare stimulation in both groups had no significant differences compared with before operation(P>0. 05).?CONCLUSION:LASIK with femtosecond laser can get a better visual quality than LASIK with microkeratome.
4.Medical image retrieval based on wavelet transform texture analysis.
Li-Dong WANG ; Xiao-Ying TAI ; Te-Er BA
Chinese Journal of Medical Instrumentation 2006;30(2):102-105
Content-based medical image retrieval has become a hot research topic due to the rapid increase of image database. In this paper, we present a method of medical images retrieval, based on texture analysis with Haar wavelet transform. Meanwhile, its effectiveness is compared with the method of retrieval based on co-occurrence matrix. A prototype system is implemented and the experiments show that the method has a much better effect.
Algorithms
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Computer Simulation
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Database Management Systems
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Image Enhancement
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methods
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Information Storage and Retrieval
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methods
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Numerical Analysis, Computer-Assisted
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Radiographic Image Interpretation, Computer-Assisted
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methods
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Radiology Information Systems
5.Medical image retrieval based on low level feature fusion and relevance feedback.
Quan ZHANG ; Xiao-Ying TAI ; Te-Er BA ; Jie-Yu ZHAO
Chinese Journal of Medical Instrumentation 2008;32(3):170-178
Based on chest images' features, this paper presents a retrieval method with domination color of partition, gray level co-occurrence matrix and gradient image shape invariant moments, as well as the fusion of them and the technique of relevance feedback. In Windows Vista environment with Visual C + +, a chest image retrieval prototype is implemented and the experiments have proved that this method has a better retrieval effect.
Algorithms
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Database Management Systems
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Diagnostic Imaging
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Image Enhancement
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Information Storage and Retrieval
6.Comparison of adherence ability of bacteria on the surface of rigid gas-permeable contact lens made with different materials
Ying-ming, WANG ; Xue-feng, QIAN ; Xiao-feng, ZHANG ; Wei, XIA ; Lei, ZHONG ; Zheng-tai, SUN ; Jing, XIA
Chinese Journal of Experimental Ophthalmology 2013;31(8):717-721
Background Wearing contaclenincreasethe risk of infection of the cornea.Some studieshowed the gas-permeability of materialused foconstructing corneal contaclenione of the contributing factorrelated to corneal health.Objective Thistudy wato observe the in vitro adherence ability of differenbacterito rigid gas-permeable contaclense(RGP-CL) made with varioumaterials.MethodContaclensemade with hexafocon,enflufocon opolymethyl methacrylate (PMMA) were placed into Staphylococcuaureus,Staphylococcuepidermidis,oPseudomonaaeruginosbacterial suspension(0.5 MCF) fo24 hours.The strength of bacterial adherence watested and studied by the methyl thiazolyl tetrazolium (MTT) colorimetrimethod based on absorbance (value),and the vortex method waused to calculate the colony forming units.The bactericlump formation waexamined with scanning electron microscope (SEM).ResultMTcolorimetrimethod showed thathe adherence ability of Staphylococcuaureuto hexafocon (value) wasignificantly lowethan thato enflufocon and PMMA,respectively (q=7.379,8.207,P<0.01),buno significandifference wafound in the adherence ability of Staphylococcuaureubetween enflufocon and PMM(q =0.828,P>0.05).The adherence ability of Staphylococcuepidermidito XO and enflufocon walowethan thato PMM(q =14.000,12.800,P<0.01),buno significandifference wafound between the adherence of Staphylococcuepidermidito hexafocon and enflufocon material (q =1.200,P>0.05).There wano significandifference in the adherence ability of Pseudomonaaeruginosto all three material(F=2.155,P=0.138).The vortex method presented the colony forming unitof Staphylococcuaureuto hexafocon,enflufocon and PMMwith (37.9± 1.5)×106,(49.9±2.2)×106 and (67.4± 1.6)×106,respectively,with significandifference among them (F =206.240,P<0.01),showing the lowesvalue in hexafocon,the highesvalue in PMMand middle value in enflufocon (q=11.650,28.640,16.990,P<0.01),Moreover,colony forming uniof Staphylococcuepidermidito hexafocon,enflufocon and PMMwa(7.9 ± 1.3) × 106,(10.5 ± 1.5) × 106,(11.2 ±1.2) × 106,respectively.And thaof hexafocon walowethan one of the PMMmaterial (q =5.060,P<0.05).No significandifference wafound between hexafocon and enflufocon nobetween hexafocon and PMM(q =3.290,1.770,P>0.05).In addition,the resultthacorresponded to the vortex method were seen in the MTcolorimetriassay (F =0.232,P =0.799).SEM examination showed dispersed population of Staphylococcuaureuand Staphylococcuepidermidion the surfaceof hexafocon and enflufocon;while much more Staphylococcuaureuand Staphylococcuepidermidiadhered on the surface of PMMA,forming net-like appearance.Conversely,high numbeof Pseudomonaaeruginoswaseen on the surface of all three materials,withounoticeable differencein the bacterial shape and quantity on each of the material.ConclusionThe adherence ability of bacterito PMMistrongethan thaof hexafocon and enflufocon,and gas-permeable material of RGP-CL doenoimpacthe adherence ability of bacteria.
7.Determination of metanephrine and normetanephrine by high-performance liquid chromatography with electrochemical detector and its diagnostic application for pheochromocytoma
Zhe LIU ; Tao YANG ; Ying XIN ; Xiao-Dong SONG ; Yu ZHANG ; Chan-Na ZHANG ; Ru-Tai HUI ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To establish high-performance liquid chromatography with electrochemical detector(HPLC-ECD) method for the determination for metanephrine and normetanephrine in 24 h urine, and provide a superior test for the diagnostic of pheochromocytomas over plasma/urine catecholamine.Methods MCX solid-phase cartridge was used for extraction of metanephrine and normetanephrine,HPLC-ECD was used for their measurements.The intra-assay CVs,interassay CVs and recoveries of metanephrine and normetanephrine were also calculated.104 hypertensive patients without pheochromocytomas and 5 pheochromocytomas patients were selected in this study.The concentrations of metanephrine and normetanephrine were compared with the plasma and 24h urinary catecholamines concentrations.Results The intra-assay CV,inter-assay CV and recovery of metanephrine were 5.9%, 7.5%,91.1% respectively;the intra-assay CV,inter-assay CV and recovery of normetanephrine were 6.3%,6.6%,88.5%,respectively.The MN,NMN,plasma CA and urine CA of all pheochromocytomas patients were positive.MN and NMN were negative in controls,while plasma CA and urine CA are false positive in 15 patients and 14 patients in controls,respectively.Conclusions The study establish a fast and accurate method for quantification of metanephrine and normetanephrine in 24 h urine by HPLC-ECD.These findings also prove that it is the best biochemical assays for pheochromocytomas at present.
8.Association of the T1270533G polymorphism in GSTM1 gene coding region with susceptibility to nasopharyngeal carcinoma in a Chinese population.
Ying HE ; Gang-qiao ZHOU ; Xin LI ; Xiao-jia DONG ; Kai-tai YAO
Journal of Southern Medical University 2009;29(1):36-39
OBJECTIVETo explore the association of T1270533G polymorphism in the glutathione S-transferase M1 (GSTM1) gene with the susceptibility to nasopharyngeal carcinoma (NPC) and clinical phenotype of NPC in Chinese population. METHDOS: The genomic DNAs were obtained from 27 Chinese subjects, and the single nucleotide polymorphism (SNP) in all the exons and relevant intron-exon boundaries of GSTM1 were determined by PCR and direct sequencing. A case-control study was performed to analyze the SNP site T1270533G (the rare allele frequency is 22.2% in Chinese population) in the coding region by means of tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and sequencing.
RESULTSequence analysis identified 29 SNPs in GSTM1 gene, among which 13 SNPs presented high linkage disequilibrium with each other. No obvious relations were found between the variation in the coding region T1270533G and the clinical phenotype of NPC (RR=0.170, 95% CI =0.95-0.306 for TT homozygotes).
CONCLUSIONThe missense mutation in the coding region T1270533G of GSTM1 gene that causes an amino acid change does not affect the detoxification function of GSTM1, and the T1270533G polymorphism does not have apparent relations to NPC susceptibility in Chinese subjects in Guangdong Province.
Adult ; Aged ; Base Sequence ; Carcinoma, Squamous Cell ; genetics ; China ; Female ; Genetic Predisposition to Disease ; genetics ; Glutathione Transferase ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; genetics ; Polymorphism, Single Nucleotide ; Young Adult
10.Construction of pcDNA3.1AM and expression of adrenomedullin in mammalian cells.
Xiao-Fang WANG ; Ying SHAO ; De-Zhi TIAN ; Tai YAO ; Li-Min LU
Acta Physiologica Sinica 2003;55(1):71-74
The newly discovered endogenous vasodilating and diuretic peptide adrenomedullin (AM) was considered to be of attractive value in clinical treatment of hypertension and congestive heart failure. In order to explore the treatment of cardiovascular diseases by expressing AM in vivo, AM cDNA was inserted into mammalian expressing vector pcDNA3.1, and in vitro expression of AM was carried out in cultured K(562) cell line. AM mRNA was amplified by RT-PCR from the total RNA isolated from the adrenal glands of rats and was inserted into pcDNA3.1 vector to form pcDNA3.1AM, the recombinant pcDNA3.1AM was then transferred into cultured K(562) cell line by liposome. The expression of AM in pcDNA3.1AM transferred cell was identified by RT-PCR and dot immunoblot assay. The results demonstrated that there were AM mRNA in the pcDNA3.1AM-transferred K(562) cell line and AM peptides in the culturing medium, indicating that the recombinant pcDNA3.1AM vector can express AM in mammalian cell line.
Adrenomedullin
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biosynthesis
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genetics
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Animals
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Genetic Vectors
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genetics
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Humans
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K562 Cells
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RNA, Messenger
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biosynthesis
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genetics
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Rats
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Real-Time Polymerase Chain Reaction
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Recombinant Proteins
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biosynthesis
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genetics
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Transfection