BackgroundIt has been well-known that integrin linked kinase(ILK) plays an important role in the pathogenesis of neorascularization.But there are few researches to elucidate the relationship between ILK and retinal nevascularization. Objective This study was to explore the expression and significance of ILK on retinal neovascularization and new vessels regression in the oxygen-induced retinopathy(OIR) mouse model. Methods One hundred and twenty-eight 7-day-old C57BL/6J mice were randomly divided into the normal group and model group.The mice were rose in(75±2)％O2 environment with mother mice together for 5 days and then in normal environment for 5 days to establish the OIR models.The mice in normal group were rose in the normal environment for 21 days.The 17-day-old mice were sacrificed and retinal sections were prepared for histopathological examination and the numbers of the cellular nuclei of vascular endotheliumbreaking retinalinternal limiting membranewere calculated.Retinal sections and sheeting were prepared in 12,14,17 and 21 -day-old mice to examine the formation and regression of new blood vessel using ADP histochemistry staining,and then immunochemistry,real-time PCR and Western-blotting were used to detect the expressions of ILK and its mRNA in retina. ResultsThe numbers of the cellular nuclei of vascular endothelium breaking retinal internal limiting membrane were (45.64 ± 12.17 )in OIR group,and those of the normal group were( 0.35±0.14 )with a significant difference between two groups (t =22.85,P＜0.05).Retinal new blood vessel appeared in 14-day mice,and peaked in 17-day mice and then regression in 21-day mice.ILK protein was expressed mainly in retinal ganglion cell layer,inner nuclear layer,inner plexiform layer and photoreceptor layer.Real-time PCR showed that ILK mRNA expressing levels in retina in model group were( 1.00±0.22),(1.85±0.17),(1.58±0.43) and(1.53±0.36) respectively in 12-,14-,17- and 21-day mice.Westernblotting determined that the A value of the ILK/β-actin was increased in 12-,14-,17-day mice and decreased in 21-day mice,and the A values were significantly higher in model group than the control group in various aged mice ( t =2.97,P＜0.05 ;t =11.88,P＜0.01 ;t =16.84,P＜0.01 ;t =13.00,P＜0.01 ). ConclusionsThese results indicate a space-time corresponding relation between the expression of ILK and retinal neovascularization.The obvious positive expression of ILK may be highly correlated with retinal neovascularization.

Background Fibrin glue has been utilized to adhere the graft during the pterygium excision with conjunctival autografts.Several relevant clinical randomized controlled trial(RCT) and retrospective studies have been published abroad,but the samples for its effectiveness and safety issue of fibrin glue and suture are still underinvestigation.Objective Current study was to quantificationally assess the efficacy and safety of fibrin glue versus sutures in the application of pterygium excision with conjunctival autografts.Methods Based on established search strategy,a computerized literature search was conducted to identify all citations concerning the RCT for effectiveness and safety evaluation of fibrin glue and suture for the graft fixation during the pterygium excision with conjunctival autografts from MEDLINE ( from 2000 to October,2010 ),EMbase ( from 2000 to October,2010 ),Cochrane Central Register of Controlled Trials ( Issue 4,2010 ),CBMdisc ( 2000 to October 2010 ),CNKI ( 2000 to October 2010 ),and the relevant conference proceedings and references searched by hand was performed as supplement.The included literature was scored with Jadad table.The Cochrane Collaboration' s RevMan 5.0 software was used for the test of heterogeneity or test for overall effect.The effective indexes,such as operative duration,recurrence rate and complication,were evaluated by Meta analysis.Results Six RCTs involving 401 eyes of 377 participants were identified.These literatures were published with English in 2004-2010 from China,New Zealand,Sweden,Israel,Turkey and Malasia and the Jadad scores were 4-5.The quantitatively analysis revealed that fibrin glue appeared to short the operative time compared with suturing method (MD =14.23 ;95％ CI:- 16.18- 12.29;P=0.00) and drop the rate of recurrence ( RR =0.49,95％ CI:0.26 -0.95 ; P =0.03 ).No significant differences were found in the rate of postoperative graft dehiscence or absence (RR =3.41,95 ％ CI:0.85-13.68;P =0.08 ).Conclusions Fibrin glue shows the good effectiveness and easy application during the pterygium excision with conjunctival autografts.Long-term follow-up of multi-central RCTs with a larger number of cases are still needed to support this conclusion.

Background Sweep pattern visual evoked potential (SVEP) is an objective method of visual test.There is a clear correlation between SVEP acuity and subjective vision,but they are not identical.Recent studies showed that new regression method can improve the accuracy of SVEP acuity. Objective This trial was to investigate and compare the outcome between amplitude-spatial frequency (A-SP) regression method and amplitudelogVA (A-logVA) regression method in extrapolating the SVEP acuity.Methods SVEP was recorded in 113 eyes of 64 subjects using GT-2000 ( Guo Te,China) with the gratings of 10 different spatial frequency from 0.99 to 12.89 cpd as stimulus.The 1 13 eyes included cataract,glaucoma,corneal disease,optical neuropathy,retinal disease,ocular trauma,refractive error and normal eyes.The correlation were analyzed of SVEP acuity,decimal visual acuity and LogMAR visual acuity.The response were averaged and DFT on the monitor display.SVEP acuity was calculated by extrapolating 0 response amplitude.Results The correlation indices of decimal visual acuity curves obtained by the A-logVA function was 0.663,and that obtained by the A-SP function was 0.705.The positive correlation was seen between subjective decimal visual acuity and A-logVA decimal visual acuity (r =0.540,P＜ 0.01 ) and between subjective decimal acuity and decimal acuity calculated by the A-SP regression method (r=0.620,P＜0.01 ).SVEP decimal acuity calculated by the A-SP function regression method was significantly different from the that calculated by the A-logVA function regression method (Z =-8.688,P＜0.01 ).And the correlation indices of LogMAR visual acuity curves obtained by the A-logVA function was 0.733 and that obtained by the A-SP function was 0.715.The positive correlation was found between the subjective LogMAR acuity and that calculated by the A-SP regression method (r=0.700,P＜ 0.01 ) and between the subjective LogMAR acuity and LogMAR acuity calculated by the A-logVA regression method (r=0.710,P＜0.01 ).SVEP LogMAR acuity from A-SP function regression method was significantly different from the LogMAR acuity from A-logVA function regression method (Z=-8.748,P＜0.01 ).No significant differences of VA LogMAR were found in gender,eyes,type of disease and age(x2 =2.171,P=0.338;x2 =0.976,P=0.614;x2 =6.032,P=0.420;x2 =14.720,P=0.257 ).Conclusions SVEP can obtain the visual outcome in human.The amplitude-logVA function regression method is more accurate in extrapolating SVEP acuity.

Objective To find out the prevalence of bullying behaviors among urban middle school students in China. Methods An anonymous questionnaire survey was conducted among middle school students selected by multiply stage cluster random sampling in 18 provinces in China. 187 328 self- administration questionnaires were finished by students from grade 7 to 12 in urban middle school(male,86 472; female, 91 106), in which 177 578 were valid. Results Bullying was common in urban middle school students in China. Multiple bullying behavior also existed. About 66.1% of boys and 48.8% of girls suffered from one or more kinds of bullying; 8.1% of boys and 2.9% of girls suffered from four or more kinds of bullying. Boys were more likely to be bullied than girls. Malicious teasing was the most common bullying behavior(43.2%), followed by sexual bullying behavior(27.0%). In addition to malicious teasing and sexual bullying, the prevalence of other types of bullying declined when the grade was increasing. Residential students were more likely to be maliciously teased, excluded or isolated and sexual bullied than non-residential students. Students from single-parent or recomposed-families were more likely to be bullied than other students. Male and ordinary school students, students living with single or step-parents were more likely to suffer multiple bullying behaviors at the same time. Conclusion The suggesting among students was associated with personal, familiar and social factors, mobilize more social forces and comprehensive actions to be taken to prevent bullying among students.

Animals ; Carcinogens ; toxicity ; Female ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Sensitivity and Specificity ; Urethane ; toxicity

Objective To investigate the waveform of the first-order kernel and second-order kernel of muhifocal electroretinogram stimulated with light emitting diode(LED).Design Prospective,noncomparative,interventional case series.Participant 18 subjects(18 eyes)who had been accepted the mfERG test.Method The patients were devided into two groups,they accepted the muhifocal elec- troretinogram(mfERG)stimulated with cathode ray tube(CRT)and LED using the Roland RETI Scan3.15 system.The first-order kernel or the second-order kernel was analyzed.The stimulation time of LED were changed from 1.7ms to 16.7ms.Five different stimulation time of LED in this study were 1/10(1.7ms),3/10(5ms),5/10(8.3ms),7/10(ll.7ms)and 10/10(16.7ms).Main Outcome Measure The summed responses were observed.The waveform,amplitude and implicit times of mfERG summed response were analyzed.Result The waveforms of the first-order kernel stimulated by LED were similar to those of CRT.In the second-order kernels of mfERG,the wave- forms were obviously different from those stimulated by LED and CRT.The P1 wave stimulated by CRT was sharp,but the P1 wave of LED was broad.The N2 wave of LED was deeper.The amplitude of N1 wave and P1 wave were increased,and their implicit times pro- longed with the stimulation times prolonging.Conclusion In the first-order kernel of mfERG,the waveform of the summed response stimulated by LED was similar to that of LED.In the second-order kernel of mfERG,the waveform stimulated by LED was more com- plicated,may be there were more inner retina information.(Ophthalmol CHN,2006,15:351-355)

Background Previous studies showed that after herpes simplex virus-1 (HSV-1) infection of the cornea,matrix metallo proteinase-2 (MMP-2) (produced by corneal cells and corneal epithelial cells) plays an important role in the development of HSK.Focal adhesion kinase (FAK)plays an important role on the expression,release and activation of MMPs.This study explored the expressions of MMP-2 and FAK,which induced by HSV-1 infected human corneal epithelial cells.Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells.Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection.The expression of MMP-2 and FAK were detected by reverse transcription PCR (RT-PCR),Western blot,immunohistochemical method and immunofluorescence method at 2 hours,20 hours and 40 hours after infection.Results At 2 hours,20 hours and 40 hours after infection,the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells (MMP-2 mRNA:Ftime =0.968,P=0.436 ; Fgroup =47.649,P =0.000 ; Fi ion =0.757,P =0.536.FAK mRNA:Ftime =0.658,P =0.631 ; Fgroup =35.182,P=0.000;Finteraction =1.386,P=0.137).Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours (P＞0.05),but the expression levels of infected cells were significantly increased at 20 hours and 40 hours (both at P ＜ 0.05).Immunohistochemistry results showed that longer infection time was associated with an increased number of cells staining for MMP-2,FAK and p-FAK.Conclusions At the initial stage of HSV-1 infected,p-FAK plays an important role in the process of virus invading and MMP-2 activation.

Objective:To express recombinant protein mIL-21-hIgGFc in 293E cells,and investigate its effect on CD8+T cell.Methods:Total RNA was extracted from the mouse spleen cells ,and then IL-21 gene was amplified by RT-PCR and inserted into expression vector PTT3-hIgGFc.PTT3-mIL-21-hIgGFc were transfected into 293E cells by calcium phosphate method.The supernatants were collected at 48 hours and 72 hours and concentrated by MOLLIPORE Labscale TM TFF system ( 5 kD membrane ).The mIL-21-hIgGFc fusion protein was purified with HiTrap TM Protein G column.The protein was quantified by SDS-PAGE and ELISA.The biological activity of the protein was determined by detecting the change of the phenotypes of CD 8+ T cells treated with the protein.Results: The constructed recombinant plasmid PTT 3-mIL-21-Fc was confirmed by sequencing.PTT3-mIL-21-Fc was transfected into 293E cells,mIL-21-Fc protein in culture supernatant was collected after 48 hours and 72 hours.The protein in cell su-pernatant reached a concentration of 787 ng/ml which was determined by ELISA.The protein was purified by Protein G chromatography column.P1A-specific T cells were treated with mIL-21-hIgGFc, and found that the CD44low CD62Lhi CD8+ population increased compared to the control.Conclusion:We built PTT3-mIL-21-hFc recombinant plasmid, expressed mIL21-hFc fusion protein in 293E cells,and purified by Protein G column.By treating mIL-21-hFc ,the antigen-primed CD8+T cells prefer to differentiate into CD44low CD62Lhi CD8+T cells which had been reported as a memory stem phenotype .This protein may be used to improve the effectness of adoptive T cell cancer therapy.

AIM: To investigate the time - effect relationship between the expression of rhodopsin and recoverin and photoreceptor damage induced by N - nethl - N -nitrosourea ( MNU) .
METHODS: Thirty-six 7-week old Sprague-Dawley ( SD ) rats were intraperitoneally injected with MNU ( 60mg/kg ) and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection ( 6 per group) , respectively. As a control, six rats were injected with phosphate buffer saline (PBS) 5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling ( TUNEL ) and transmission electron microscope ( TEM ) in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.
RESULTS:The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak ( 29. 7% ±2.3%) at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection (P<0. 05), and extremely significant difference comparing to control group after 12h ( P<0. 01 ); while recoverin dramatic declined at 12h, and extremely significant difference after 24h (P<0. 01).
CONCLUSION:60mg/kg MNU intraperitoneally injection one - time may specifically induce photoreceptor apoptosis, The mechanism of down - regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.

Background Oxygen-induced retinal neovascularization is the main pathological basis for many retinal vascular diseases.Research showed that light exposure at night can suppress retinal neovascularization in oxygen-induced retinopathy(OIR),but there were few reports discussing its effect on ROP.Objective This study aimed to observe the effect of light exposure at night on retinal neovascularization in an OIR mouse model.Methods Sixty-four newborn C57 BL/6J mice were randomly divided into four groups,with 16 mice for each group.OIR models were established by rearing the newborn C57BL/6J mice with their mothers in a(75±2)％ oxygen environment from postnatal day 7(P7)to Pl2,and then transferred to room air.In the OIR model group,the environmental illumination level was the same as the normal control group,and the model mice were exposed to 100 lx light at night in the OIR+ light exposure group.In the simple light exposure group,normal mice were reared in room air and were exposed to light at night from P12 to P17.All the mice were sacrificed on P17,and retinal flat mounts were prepared to assess the oxygen-induced changes of retinal vessels using the adenosine diphosphatase(ADPase)histochemical technique.The amount of proliferative neovascularization was quantified by counting the number of endotheliocyte nuclei in new vessels extending from the retinal inner limiting membrane into the vitreous in ocular cross-sections.The expression of the vascular endothelial growth factor(VEGF)protein was detected by immunohistochemistry.Real-time PCR analysis was performed to examine the expression of VEGF mRNA.The rearing and usage of the animals complied with the Statement of ARVO.Results Less free-vascular areas and new blood vessels were seen in the OIR+light exposure group compared with the OIR model group.On day 17 of the mouse life,the number of the endotheliocyte nuclei in new vessels extending from retinal inner limiting membrane were 0.97±0.83,1.00±0.72,38.57±5.01 and 16.92±3.39 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing significant differences among them(F =78.767,P =0.000).The number of nuclei in the OIR+light exposure group were less than that of the OIR model group(t=20.446,P＜0.01).Immunochemistry showed that the expression of VEGF in retina was weaker in the OIR+light exposure group than the OIR model group.The relative expression values of VEGF mRNA were 1.00±0.00,0.94±0.07,2.08±0.50 and 1.43±0.21 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing a significant difference (F=11.268,P =0.003),where the VEGF mRNA level in the OIR+light exposure group was lower than that of the OIR model group(t =20.163,P＜0.05).Conclusions Light exposure at night can weaken retinal neovascularization in OIR mice