RNA-seq is a transcriptome analysis method using deep sequencing technology, which uses high-throughput next-generation sequencing technology to investigate, characterize and quantify transcriptome.It can quickly and comprehensively detect almost all transcripts and gene sequences of specific cells in specific tissue of a species, and has been widely used in basic research, clinical diagnosis and drug development and other fields.Its measurement accuracy has been confirmed comparable to microarray and quantitative polymerase chain reaction by several studies.RNA-seq technology can accurately detect mRNA and non-coding RNA which play important roles in the pathogenesis of diabetic retinopathy (DR). Using this technology to investigate the gene regulation and molecular mechanism of DR is helpful to find biomarkers for early diagnosis and prognosis of DR, to comprehensively and systematically study and analyze the molecular mechanism of specific biological process, and to find new therapeutic targets.The application of RNA-seq will be of great significance for the basic research and clinical treatment of DR.In this article, RNA-seq technology was comprehensively and systematically expounded from three aspects, which were the advantages of RNA-seq technology, the selection of sequencing platform as well as library preparation and data analysis.The progress of this technology in DR research was summarized and analyzed.

Objective To observe the therapeutic efficacy and postoperative complications of combined surgery and sequential surgery in treating proliferative diabetic retinopathy (PDR) and cataract.Methods The cilinical data of 59 patients (66 eyes) with PDR were retrospectively analyzed.The patients were divided into combined-surgery group and sequential-surgery group.Combined surgery was performed on patients with obvious lens opacity which was an obstacle to the ocular fundus surgery,while sequential surgery was performed on the patients with transparent lens or lens with light opacity on which ocular fundus surgery could be performed.Lens excision in cataractopoiesis could only be performed when the state of ocular fundus was stable and cataractopoiesis was the main cause of vision damage.A total of 28 patients (32 eyes) in combined-surgery group underwent vitrectomy combined with phacoemulsification and IOL implantation;31 patients (34 eyes) in sequential-surgery group underwent vitrectomy,lens excision and IOL implantation.Corrected visual acuity (BCVA) and complications were observed.The mean follow-up period was (25±8.5) months.The measurement data were analyzed with t test and enumeration data with 2 test.Results In combined-surgery group,visual acuity improvement was achieved in 27 eyes (84.4%),remained unchanged in 2 eyes (6.3%) and decreased in 3 eyes (9.4 %).In sequential-surgery group,visual acuity improved in 26 eyes (76.5%),remained unchanged in 2 eyes(5.8%)and decreased in 6 eyes (17.7%.).However,anterior chamber fibrin exudation occurred in 4 eyes in the combined-surgery group and no eyes in the sequential-surgery group;compared with each other,the difference is statistically significant (χ2=4.524,P=0.033).Conclusion Combined surgery and sequential surgery are all safe and effective on treating PDR and postoperative complications have no obvious correlation with the surgery procedure.

The incidence of high myopia has increased yearly, which has been an area of focus for investigation. The complications of high myopia are contributing factors to blindness,such as posterior staphyloma, chorio-retinal atrophy, choroidal neovascularization, optic nerve head and macular defects. The study of histological changes helps clarify the mechanism. With the development of modern technology such as spectral-domain optical coherence tomography and enhanced depth imaging-technique, histological changes in high myopia can be measured accurately. The purpose of this review is to provide an update on various pathologic changes, including in sclera, Choroid/Bruch's membrane, optic nerve, vitreous and retina.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma in the Western world,and is potentially curable with standard R-CHOP chemoimmunotherapy. We are now in an era that the heterogeneity of DLBCL is defined genetically and molecularly,and rational subset-specific therapeutic targets are guiding clinical trials.Primary mediastinal DLBCL is a unique clinicopathologic entity, and alternatives to R-CHOP may confer superior outcome. Rearrangement of the myc oncogene occurs in 10%of patients with DLBCL, and confers a very poor prognosis with standard R-CHOP, particularly when there is concomitant rearrangement of bcl-2, a condition referred to as double-hit DLBCL. A larger subset of DLBCL demonstrates overexpression of both myc and bcl-2 by immunohistochemistry. Analyze the source of cells by gene expression profile, immunohistochemistry algorithms,or a novel Lymph2Cx platform,provides prognostic information, and guides therapeutic decisions in both relapsed and de novo disease. This article reviews latest research presented at the 57th American Society of Hematology (ASH) annual meeting on the definition of specific subsets of DLBCL and selection of subtype-specific treatment,including novel approaches under investigation. Understanding these key features of the pathology report, and limitations of these assays defining subsets of DLBCL, allows for a precision medicine approach to this disease.

ObjectiveTo investigate the effect of cholinesterase inhibitor on endotoxin-induced brain injury in rabbits.Methods Twenty-one healthy male rabbits were randomly assigned into three groups ( n ＝ 7each):group sham operation (group S),lipopolysaccharide (LPS) group and cholinesterase inhibitor (tacrine hydrochloride,THA) group.LPS 200 μg/kg was intracerebroventricularly injected in LPS group,LPS 200μg/kgand tacrine hydrochloride 150 μg/kg were injected in THA group,while same volume of normal saline was injected in S group.Then blood and tissue samples were collected in different groups after 4 hours.Nuclear factor-kappa B (NF-κB) p65 activity of brain tissues was determined by using Western blot analysis.Tumor necrosis factor-alpha (TNF-α) levels in plasma,cerebrospinal fluid and brain tissues were measured using enzyme linked immunosorbent assay.The brain tissue's myeloperoxidase (MPO) activity and the ratio of wet to dry weight (W/D) were also analyzed.ResultsAs compared with S group,TNF-α level in plasma,cerebrospinal fluid and brain tissues,NF-κB p65 level,MPO activity and W/D ratio increased in LPS and THA groups (P < 0.05).When compared with LPS group,TNF-α level in plasma,cerebrospinal fluid and brain tissues,NF-κB p65 level,MPO activity and W/D ratio decreased in THA group ( P < 0.05 ).ConclusionCholinesterase inhibitor can attenuate the endotoxin-induced brain injury through inhibiting local inflammatory responses.

Objective To evaluation clinical efficacy and complications characteristics of vitrectomy on type 1 diabetes of diabetic retinopathy.Methods This was a retrospective review study.From January 2010 to January 2016,18 cases (28 eyes) under the age of 30 diagnosed as type 1 diabetes of diabetic retinopathy were treated in our hospital.All the patients were performed vitrectomy.The postoperative visual acuity and complications in and after operation were observed.Results The follow-upperiod was 6-72 months with the average of 43.0 months.Postoperative visual acuity improved or remained unchanged in 23 eyes (82.1％),and decreased in 5 eyes(17.9％).Our primary anatomic success rate was 92.3％.Complication in operation was iatrogenic retinal tear in 3 eyes.Postoperative complications consisted of anterior chamber fibrin exudation in 9 eyes,hyphema in 5 eyes,recurrent vitreous hemorrhage in 2 eyes,retinal detachment in 3 eyes,rubeosis of iris in 5 eyes and neovascular glaucoma in 4 eye.Conclusion For young patients of type 1 proliferative diabetic retinopathy,vitrectomy can restore retinal anatomical structure and function.Panrentinal photocoagulation and preoperative anti-VEGF injections in vitreous is necessary with serious neovascularization-related complications.

Objective To investigate the clinical efficacy of posterior lumbar interbody fusion and internal fixation for the treatment of lumbar spinal stenosis with redundant nerve roots (RNRs).Methods A retrospective case series study was performed on the clinical data of 23 cases of lumbar spinal stenosis with RNRs from January 2009 to December 2014.This study involved 10 males and 13 females,with an average age of 48.4 years(range,38-58 years).The course of disease averaged 28.1 months(range,2 months-7 years).There were seven patients with single segment of stenosis,12 patients with two segments of stenosis,and four patients with three segments of stenosis.RNRs diagnostic criteria:in the sagittal section of the MRIT2 image of lumbar spine,the cauda equina nerve root in the dural sac was circular,tortuous and twining.MRI information was collected and analyzed by three double-blind radiologists,respectively.RNRs were determined based on the unanimous consent of all the three doctors.All patients underwent posterior discectomy,spinal canal decompression,interbody fusion,and internal fixation after admission.The operation time,intraoperative blood loss,and postoperative complications were recorded.The visual analogue scale (VAS),Oswestry disability index (ODI),and Japanese Orthopedic Association (JOA) score were used to evaluate the back pain and leg pain before operation and 3 months after operation.Fischgrund standard was used for the overall efficacy evaluation.Results All patients were followed up for 12-30 months (mean,23.3 months).The operation time was (130.0 ± 23.2) minutes,and the intraoperative blood loss was (513.0 ± 165.0) ml.MRI was reviewed 1 week after operation,and the result showed that redundancy of cauda equina disappeared.Preoperatively,the scores of back pain VAS,leg pain VAS,ODI and JOA were (6.3± 0.8) points,(6.8 ±0.9)points,(46.7±2.5)points and (10.3 ±2.8)points,respectively.At 3 months postoperatively,the scores were (1.4 ± 0.5) points,(1.8 ± 0.7) points,(11.9 ± 2.1) points,and (25.3 ± 1.8) points,respectively (P ＜ 0.05).In terms of the efficacy,17 cases were excellent,four good,and two fair,with an excellent and good rate of 91％.At 12 months after operation,the CT scan showed no screw rupture or cage dislocation,with bony union seen between the vertebrae.Conclusion Posterior lumbar interbody fusion and internal fixation can help relieve the back and leg pain and eliminate redundant nerve roots in patients with lumbar spinal stenosis with RNRs.

Intravitreal injections of anti-vascular endothelial growth factor (anti-VEGF) drugs,including monoclonal antibodies (such as bevacizumab and ranibizumab) and fusion protein agents (such as aflibercept and conbercept) have been proven to be effective in the treatment of wet age-related macular degeneration (wAMD).However,there are still some patients with poor efficacy,such as no response to initial treatment or poor response,and even relapse during the course of treatment.In view of the different targets and molecular characteristics of anti-VEGF drugs,the switch of anti-VEGF drugs and the adjustment of delivery pattern,dosages and intervals have been the strategies to cope with the poor efficacy in clinic.However,there are some differences in the results of current studies.Overall,the recovery of retinal anatomical outcome achieves more benefits,and it is relatively difficult to improve visual acuity.To determine which regimen would get the biggest benefits,a large number of randomized controlled clinical trials and long study period will be needed.

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) on hydrogen peroxide (H2O2) induced apoptosis of retinal pigment epithelial (RPE) cells in vitro.Methods RPE cells were cultured and divided into a normal group,normal+H2O2 group,Vec+H2O2 group,PSF+H2O2 group according to the experimental design.Overexpression of PSF in RPE cells were achieved by pEGFP-PSF plasmid transient transfection into RPE cells,then RPE cells were exposed to H2O2.The morphological changes were observed by hematoxylin-eosin (HE) staining and Live/Dead staining while the survival rate of cells was detected by MTT assay.The effect of PSF on H2O2-induced RPE apoptosis was analyzed by Cell Death Detection ELISA kit.Meanwhile,intracellular reactive oxygen species (ROS) level was detected by using DCFH-DA method.Results Overexpression of PSF could effectively alleviate the morphological changes induced by H2O2 stimulation shown by HE staining,and effectively reduce dead cells number shown by Live/Dead staining.After H2O2 stimulation,the survival rate,apoptosis rate and ROS production level in PSF overexpression group were 0.68± 0.12,0.44± 0.08 and 18 616± 3 382.54 respectively,showing significant difference in comparison with the vector plasmid group and normal group (P＜0.05).Conclusion PSF overexpression plays a protective role in H2O2-induced apoptosis by inhibiting the production of ROS in RPE cells.

Objective To observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.Methods Cultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments.The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group.The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy.VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment,and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq.Now with biological big data obtained as a basis,to analyze the differentially expressed genes (DEGs).And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.Results The gene expression profiles of the two groups of cells were obtained.Through analysis,328 DEGs were found,including 194 upregulated and 133 downregulated ones.The functions of DEGs were influenced by regulations over molecular biological process,cellular energy metabolism and protein synthesis,etc.Among these genes,SI,PRX and HPGD were related to protein synthesis,BIRCT to cellular apoptosis,and ABLIM 1 and CRB2 to retinal development,and ABCG 1,ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid.GO enrichment analysis showed that DEGs mainly act in three ways:regulating biological behavior,organizing cellular component and performing molecular function.Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway,and Notch,mitogen-activated protein kinase,transforming growth factor (TGF)-β and Wnt signal pathways.Among them,the gene expression in TGF-β signal pathway attracts most attention,where the DEGs,such as CAMK2B,COL3A1,CYGB,PTGER2 and HS6ST2,among others,were closely related to fibrosis process.Conclusion The anti-VEGF drugs may enhance the expression ofCAMK2B,COL3A1,CYGB,PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.