Diabetic macular edema (DME) is one of causes leading to visual acuity loss in the diabetic population.In recent years,great progress has been achieved in the treatment for DME.While laser photocoagulation and pars plana vitrectomy remain the standard of care,a new wave of methods,including intravitreal corticosteroid and anti-vascular endothelial growth factor (anti-VEGF) medications,is emerging and shows the preliminary safety and efficacy profile for DME by a number of clinical trials.This article introduce the definition,classification,pathogenesis and the different treatment methods for DME,so to aid ophthalmologists in evaluating the most appropriate and up-todate management options in managing DME.Particular attention is paid to summarize recent peer-reviewed literatures of ranibizumab (Lucentis).

Optical coherence tomography angiography (OCTA) is a noninvasive angiography technique developed in recent years.Without using contrast medium,this technology can quickly and safely get retinal vascular images with relatively high-resolution.It has been widely used in the diagnosis and management of eye diseases,especially in the vascular diseases,such as diabetic retinopathy (DR),age-related macular degeneration (AMD),retinal central/branch venous obstruction (CRVO/BRVO),choroidal neovascularization(CNV),etc.This article reviews the clinical application of OCTA in diabetic retinopathy.

Objective To investigate the expression of Ezrin in gastric cancer tissues and its clinical significance.Methods Gastric cancer tissues and adjacent normal gastric tissues from 60 patients with gastric cancer were collected from June 2008 to May 2009 at the Southwest Hospital.The mRNA and protein expressions of Ezrin were detected by using the reverse transcription polymerase chain reaction and western blot.The relationship between Ezrin and the gender and age of patients,and tumor differentiation,pathological staging,depth of invasion and lymph node metastasis was analyzed.All data were analyzed using the t test,chi-square test and Spearman rank correlation.Results The Ezrin mRNA expression level was increased in 33 (55％) cases of adjacent normal gastric tissues and 21 (35％) cases of gastric cancer tissues; the Ezrin protein expression level was increased in 45 (75％) cases of adjacent normal gastric tissues and 22 (37％) cases of gastric cancer tissues.The mRNA and protein expressions of Ezrin in the normal adjacent gastric tissues were 1.30 ± 0.04 and 3.57 ± 0.45,respectively,which were significantly higher than 0.53 ± 0.36 and 0.96 ± 0.18 in the gastric cancer tissues ( t =5.309,22.617,P ＜ 0.05 ).The mRNA expression of Ezrin was positively correlated with the protein expression of Ezrin (r =0.602,P ＜ 0.05 ).The mRNA and protein expressions of Ezrin were related to the pathological stages,depth of invasion and state of lymph node metastasis (x2 =6.41,6.49,4.62; 5.40,8.87,4.12,P ＜ 0.05),but not to the gender,age and tumor differentiation (x2 =0.50,0.07,1.07 ; 0.01,1.16,1.96,P ＞ 0.05).Conclusion The mRNA and protein expressions of Ezrin are significantly decreased in the gastric cancer tissue,which might be responsible for genesis,development and metastasis of gastric cancer.

A biomedical entity association evolution network was constructed by mining the implicit associations in PubMed-covered literature, which can help scientific researchers to form new scientific hypotheses, to analyze the topological features of associated network, to study the scientific literature-enriched knowledge structure, associa-tions, development rules, to introduce new visual angles and methods for literature-based knowledge discovery, and to improve the knowledge discovery efficiency.

Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

AIM: Using optical coherence tomography angiography (OCTA) to observe the changes and clinical significance of macular blood flow density in patients with diabetic retinopathy (DR).METHODS: Totally 47 eyes (28 patients) with diabetic retinopathy (DR) were enrolled in the DR group.According to the international clinical grading criteria of diabetic retinopathy, 30 eyes (19 patients) with non-proliferative diabetic retinopathy were classified as the NPDR group, and 17 eyes (11 patients) with proliferative diabetic retinopathy were classified as PDR group.A total of 46 (27 subjects) healthy eyes with matched age were enrolled in the control group.All the subjects underwent the 3mm×3mm scanning of macular retina by optical coherence tomography angiography (OCTA), obtaining 4 levels of macular blood flow density map.The macular blood flow density at 3 levels, including superficial retinal layer, deep retinal layer and choroidal capillaries layer, were measured.RESULTS: The macular blood flow density of superfical retinal layer, deep retinal layer and choroidal capillaries layer in DR group were 0.4963±0.0840, 0.4798±0.0801 and 0.5290±0.0528, respectively.Among them, the blood flow density of each layer were 0.5064±0.0843,0.4983±0.0766,0.5345±0.0529, respectively, for the NPDR group, and were 0.4786±0.0830, 0.4473±0.0778,0.5192±0.0526, respectively, for the PDR group.For the control group, the density of each layers were 0.5919±0.0704, 0.6301±0.0527, 0.5691±0.0169, respectively.The macular blood flow density was significantly different in the superficial retinal layer, deep retinal layer and choroidal capillary layer between the control group and the NPDR group, as well as the PDR group and the DR group (total P<0.001).Statistically significant difference was found between the NPDR group and the PDR group in the deep retina layer (P=0.029), but not in the superficial retina layer and choroid capillary layer (P=0.236, 0.268).CONCLUSION: Compared with the control group, the macular blood flow density of superficial retinal layer, deep retinal layer and choroidal capillary layer in the patients with diabetic retinopathy decreased significantly.It indicated that the macular ischemia existed in both retina and choroid.By quantitatively measurement of the macular blood flow, OCTA may be used for monitoring the progression of diabetes, and early detection of diabetic retinopathy.

The effect of paper publication time on knowledge discovery in biomedical literature was studied by analyzing the role of time factor in indirect link mining and tested using the PubMed-covered data set,which showed that recently published papers played a better role than early published papers in knowledge discovery,indicating that the time factor plays a certain role in knowledge discovery in biomedical literature and can thus improve the efficiency of knowledge discovery in biomedical literature.

Objective:To establish an LC-MS/MS method for the simultaneous determination of acetaminophen and its five metabolites in mice plasma,and investigate the metabolism of acetaminophen by using the method.Methods:Para aminobenzoic acid was used as the internal standard.The plasma samples were precipitated by methanol,and then separated on a C18 column with the mobile phase of methanol and 5 mmol·L-1 ammonium acetate buffer solution containing 0.1% formic acid (55∶45).The flow rate was 0.5 ml·min-1,and the column temperature was 25℃.An electrospray ionization source was applied and operated in a positive ion mode using MRM:APAP,-,m/z 152.0→110.0;APAP-cys,-,m/z 271.2→140.1;APAP-glut,-,m/z 457.0→328.0;APAP-NAC,-,m/z 313.4 →208.0;APAP-sulf,-,m/z 232.4→152.1;APAP-gluc,-,m/z 328.2→152.1;IS,-,m/z 138.2→120.0.Results:The method exhibited good linearity over the concentration range of 0.2-10 μg·ml-1for APAP,1.0-20 μg·ml-1 for APAP-gluc,1.0-20 μg·ml-1 for APAP-sulf,1.0-20 μg·ml-1 for APAP-glut,0.4-15 μg·ml-1 for APAP-NAC and 0.2-10 μg·ml-1 for APAP-cys (r≥0.990 0).The inter-day accuracy and precision of acetaminophen and its five metabolites were all below 15%.The average recovery was between 85% and 115%,and RSDs were all below 15%.Conclusion:The LC-MS/MS method is proved to be quick,sensitive and accurate,and suitable for the determination of acetaminophen and its five metabolites in mice plasma.

Objective To study the effect of transfection with antisense DNMT3b gene eukaryotic expression plasmid on the growth of human cholangiocarcinoma cell line QBC939,and to explore the role of DNMT3b in the cholangiocarcinoma tumorigenesis.Methods The constructed antisense DNMT3b gene eukaryotic expression plasmid was transfected into QBC939 cells using liposome.The expression level of DNMT3b protein was detected by Western blot after stable transfection.The growth curves of transfected cells and un-transfected cells were observed by MTT method respectively.The cell proliferation ability was also observed by the test of colony formation in soft agar.The alterations of the cell cycle and the apoptosis rate were detected by FCM.Results Following the transfection,the protein level of DNMT3b decreased significantly;transfection with antisense DNMT3b gene eukaryotic expression plasmid did not affect the cell growth curve of QBC939,and did not decrease the cell colony formation rate(P=0.717);transfection with antisense DNMT3b gene also did not result in cell cycle alterations or induce cell apoptosis(P=0.089).Conclusions Transfection with antisense DNMT3b gene eukaryotic expression plasmid can down-regulate the expression level of DNMT3b in QBC939.It can not affect the growth and proliferation of human cholangiocarcinoma cell line QBC939,nor alter the cell cycle and induce cell apoptosis.

Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16％)or hypoxia condition(10％ O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10％ O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16％ O2 culture group(0.16+0.02)(P＜0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10％ O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16％ O2 culture group(0.06±0.01)(P＜0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00％ in 16％ O2 culture group to 36.00％ and 46.00％ in the cells cultured by 10％ O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90％ and 46.10％,respectively,but that in 16％ O2 culture group was 9.10％,showing a statistically significant difference(P ＜ 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.