Diabetic retinopathy (DR) is one of the main causes of vision loss and irreversible blindness in the working-age population, closely regarded as the destruction of the retinal neurovascular unit (NVU). As an important component of the NVU, retinal microglia (RMG) plays a vital role in the progression of DR. In recent years, single-cell RNA sequencing (scRNA-seq) technology has emerged as an important tool in transcriptomic analysis. This latest method reveals the heterogeneity and complexity of RNA transcriptional profiles within individual cells, as well as the composition of different cell types and functions. Utilizing scRNA-seq technology, researchers have further revealed the role of RMG in the occurrence and development of DR, discovering phenotypic heterogeneity, regional heterogeneity, and cell-to-cell communication in RMG. It is anticipated that in the future, more omics technologies and multi-omics correlation analysis methods will be applied to DR and even other ophthalmic diseases, exploring potential diagnostic and therapeutic targets, providing different perspectives for the clinical diagnosis, treatment, and scientific research of DR, and truly promoting clinical translation through technological innovation, thereby benefiting patients with DR diseases.

Objective To investigate the relationship between the expression of matrix metalloproteinase(MMP)-2,MMP-9,MMP-13 and disease activity in pemphigus patients.Methods A total of 60 pemphigus patients treated in the dermatology department of the hospital from January 2021 to January 2023 were select-ed as the study group,and another 60 healthy volunteers who underwent physical examination in the same pe-riod were recruited as the control group.The levels of MMP-2,MMP-9 and MMP-13 in serum and the expres-sion of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells were compared between the two groups,and the relationship between the expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of patients with pemphigus was analyzed.Pemphigus patients were divided into acute phase group(n=22),chronic phase group(n=23)and stable phase group(n=15)according to their disease activi-ty.The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of the three groups were compared,as well as the correlation between the three indexes and disease activity.Receiver operating characteristic(ROC)curve was plotted for analysis.Results The levels of MMP-2,MMP-9 and MMP-13 in serum and the expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells in the study group were higher than those in the control group,with statistical significance(P＜0.05).The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells in severe patients were higher than those in moderate and mild patients,and the difference was statistically significant(P＜0.05).The expression of MMP-2,MMP-9 and MMP-13 and disease activity scores of mononuclear cells in acute attack group were higher than those in chronic attack group,and the differences were statistically significant(P＜0.05).The ex-pression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells were positively correlated with disease activity(rMMP-2=0.545,rMMP-9=0.592,rMMP-13=0.580,P＜0.05).ROC curve analysis showed that compared with the single diagnostic efficacy of MMP-2,MMP-9 and MMP-13 expression in peripheral blood mononuclear cells,the three combined diagnostic efficacy was better.Conclusion The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of pemphigus patients is related to the disease activity,and these three indicators can be used as reference indicators for the diagnosis of pemphigus disease activity.

AIM: To investigate the influencing factors of vault after the posterior chamber phakic refractive lens(PC-PRL)implantation for patients with super high myopia.METHODS: Retrospective case study. A total of 40 patients with super high myopia(77 eyes)who underwent PC-PRL implantation in the Haixiang Eye Hospital from January 2019 to January 2021 were selected. They were followed up for at least 2a, postoperative anterior segment parameters, such as the uncorrected visual acuity(UCVA), best corrected visual acuity(BCVA), central anterior chamber depth(ACD), anterior chamber volume(ACV), anterior chamber angle(ACA), lens thickness and vault were evaluated, and then the influencing factors of postoperative vault were analyzed.RESULTS: The UCVA and BCVA of the patients significantly improved after PC-PRL implantation(P<0.001). Average safety index(postoperative BCVA/preoperative BCVA)was 1.36±0.32, and average effective index(postoperative UCVA/preoperative BCVA)was 1.23±0.31 in 2a after surgery. The vault in 2a after surgery was correlated with preoperative ACD, ACV, ACA and lens thickness, and the preoperative ACV and lens thickness had significant impact on vault in 2a after surgery.CONCLUSIONS: The PC-PRL implantation is safe and effective in super high myopia, and it can significantly improve visual acuity. Furthermore, preoperative ACV and lens thickness are important influencing factors of postoperative vault.

Objective: To construct the full-length prokaryotic expression plasmid of the wild type of androgen receptor (AR) and the truncated body of four functional domains，and to identify the fusion protein by Western blot and electrophoretic mobility shift assay ( EMSA) ． Methods: Based on the pGEX-4T-1 vector ，the recombinant plasmids were constructed to express the full-length and functional domains of AR. IPTG was used to induce the expression of the recombinant proteins，which were isolated and purified by glutathione sepharose 4B beads under the optimized condition．The specific protein expression in the bacterial lysate and the purified protein isolated with glutathione sepharose 4B beads was identified by Western blot with AR antibody and GST labeled antibody．The purified protein was incubated with a fluorescent probe of the virus，and the complex was detected by electrophoresis in a non-denaturing gel. Results : The prokaryotic recombinant plasmids of full length and three functional domain truncated AR were successfully constructed．The recombinant clones were identified by using bacterial culture as a template，and further verified by double enzyme digestion．It showed that there were identical bands in the same sizes as the inserted fragments．The nucleotide and the amino acid sequences were aligned to the reference sequence in NCBI GenBank．The GST fusion protein，GST-AR-NTD + DBD (96 ku) and GST-AR-NTD (86 ku) were successfully induced and verified． The purified protein could be directly combined with the viral genome DNA． Conclusion The prokaryotic expression conditions of truncated AR plasmid from the same gene sequence are different．The purified AR protein can be used to understand the direct interaction mechanism between functional domains of AR and other molecules.

Objective:To observe the clinical and multimodal imaging features of eyes with acute macular neuroretinopathy (AMN) associated with the coronavirus disease 2019 (COVID-19).Methods:A retrospective study. From December 20, 2022 to January 17, 2023, a total of 29 patients (58 eyes) with COVID-19-associated AMN admitted to Department of Ophthalmology of Eye and ENT Hospital, Shanghai Medical College were included in the study. All the affected eyes underwent the best corrected visual acuity, color fundus photography, infrared fundus photography (IR), short-wavelength autofluorescence (SW-AF), near-infrared autofluorescence (NIL-AF), optical coherence tomography (OCT), and OCT angiography (OCTA). All patients were administered microcirculation-improving oral medication with 12 cases receiving adjunctive low-dose corticosteroid therapy. Follow-ups were conducted 1 to 3 months after the initial diagnosis, with a total of 19 cases (38 eyes) completing the one-month follow-up.Results:Out of the 29 cases, there were 9 males (18 eyes) and 20 females (40 eyes), all of whom experienced bilateral eye involvement. The age of the patients ranged from 12 to 47 years, with an average age of (29.9±9.5) years. The time from the onset of fever to the appearance of ocular symptoms was (2.52±2.01) days. Among the 58 affected eyes, there were 5 cases with retinal cotton wool spots, 2 cases with optic disc edema, and 1 case with parafoveal branch retinal vein occlusion. All affected eyes exhibited deep reddish-brown macular dark spots. IR revealed wedge-shaped, wedge-like, or "petaloid-like" dark areas involving the fovea and parafovea. SW-AF examination showed no obvious abnormality in 39 eyes. Weak autofluorescence dark area were consistent with IR in 19 eyes. NIR-AF examination showed spot-like or flaky self-fluorescent dark areas. OCT examination showed strong reflex lesions spreading vertically upward from the retinal pigment epithelium (RPE) layer in the macular area in the acute stage, showing typical "bean seedling" sign. OCTA revealed reduced blood flow density in the deep capillary plexus (DCP) of 50 eyes. Enface OCT displayed lesion areas that corresponded to the dark areas seen in IR. One month after the initial diagnosis, the condition improved in 18 eyes (47.4%, 18/38). Among the 5 eyes with cotton wool spots, regression of these spots was accompanied by loss of nerve fiber layer in 4 eyes. In cases with optic disc edema, the edema subsided. The "bean sprout" sign disappeared in all affected eyes, and the lesions became localized. The ellipsoid zone and/or interdigitation zone in the lesion areas were discontinuous.Conclusions:COVID-19-related AMN is characterized by distinctive features. IR fundus reveals wedge-shaped, wedge-like, or petaloid dark areas involving the fovea and parafovea. OCT displays strongly reflective lesions with vertical spread above the RPE. OCTA shows reduced blood flow density in the DCP of the retina.

According to the characteristics of short time and large amount of samples for out of hospital emergency nucleic acid detection, this study introduces an out of hospital emergency nucleic acid detection cloud platform system, which realizes the functions of rapid identification of the detected person and one-to-one correspondence with the samples, and real-time upload of the detection results to Zhejiang Government service network for quick viewing and statistics, so as to complete the task of national nucleic acid screening efficiently and accurately that we must provide information support.
COVID-19 ; Cloud Computing ; Humans ; Nucleic Acids ; SARS-CoV-2

The axon initial segment (AIS) is a specialized structure that controls neuronal excitability via action potential (AP) generation. Currently, AIS plasticity with regard to changes in length and location in response to neural activity has been extensively investigated, but how AIS diameter is regulated remains elusive. Here we report that COUP-TFI (chicken ovalbumin upstream promotor-transcription factor 1) is an essential regulator of AIS diameter in both developing and adult mouse neocortex. Either embryonic or adult ablation of COUP-TFI results in reduced AIS diameter and impaired AP generation. Although COUP-TFI ablations in sparse single neurons and in populations of neurons have similar impacts on AIS diameter and AP generation, they strengthen and weaken, respectively, the receiving spontaneous network in mutant neurons. In contrast, overexpression of COUP-TFI in sparse single neurons increases the AIS diameter and facilitates AP generation, but decreases the receiving spontaneous network. Our findings demonstrate that COUP-TFI is indispensable for both the expansion and maintenance of AIS diameter and that AIS diameter fine-tunes action potential generation and synaptic inputs in mammalian cortical neurons.
Action Potentials ; Animals ; Axon Initial Segment ; COUP Transcription Factor I ; DNA-Binding Proteins/physiology* ; Mammals ; Mice ; Transcription Factors

The authors introduced the construction of the central monitoring system of bedside monitor in a hospital, and introduced its software and hardware design scheme and function in detail. The implementation of the system guaranteed the medical safety, reduced the workload of medical staff, improved the work efficiency, and had the characteristics of low cost and practicability.

Objective To observe the alterations of vascular density of superficial or deep retinal vascular plexus and the area of foveal avascular zone (FAZ) in acute central serous chorioretinopathy (CSC) eyes. Methods This is a retrospective study including 22 patients with unilateral CSC in acute stages. The patients were divided into group A (22 affected eyes) and B (22 fellow eyes). Twenty eyes of 20 age and gender-matched healthy volunteers (group C) were enrolled in this study. All subjects underwent optical coherence tomography (OCT) angiography examinations. The observation index included signal intensity of blood flow in choroidal capillary, vessel density and FAZ on the superficial capillary plexus (SCP) and deep capillary plexus (DCP). Results The FAZ on DCP in group A was larger than that in group B and C (t=4.28, 5.57; P=0.00、0.00), but there was no significant difference of FAZ on SCP among 3 groups (t=0.28, 0.80; P=0.78, 0.43). The vessel density of SCP and DCP in group A and B were decreased than those in group C (t=-4.40, -2.91; P=0.00, 0.00). The type of choroidal capillaries dilation had two different manifestations by OCTA, which was circumscribed in 12 eyes and diffuse in 10 eyes. The subretinal fluid in macular area which presented dark chamber with weak reflection was observed by en-face OCT. Conclusions The vessel density of SCP and DCP in both eyes of acute CSC are lower than those in normal eyes. The FAZ of DCP in affected eyes of acute CSC is larger than that in fellow eyes and normal eyes. The blood flow of dilated choroidal capillary can be manifested as different morphological enhancements.

Objective To study the effect of brain‐derived neurotrophic factor (BDNF) in the process of adenosine triphos‐phate (ATP) activating BV2 microglia .Methods BV2 microglia was cultured by adding different concentrations of ATP .Then the expression level of intracellular CD11b and BDNF and the secretion level of TNF‐α in the supernatant were quantitatively deter‐mined by Western blot .BV2 microglia was treated by different concentrations of BDNF scavenger tyrosine kinase receptors B (TrkB)/Fc and incubated by ATP .The expression level of intracellular CD11b and BDNF and the secretion level of TNF‐αin the supernatant were measured .Finally adding exogenous recombinant BDNF into cultured BV 2 microglia ,intracellular changes of CD11b and supernatant TNF‐αlevels were detected .Results After adding ATP for cultivating BV2 microglia ,intracellular CD11b and BDNF expression levels and supernatant TNF‐αlevel were increased with a dose‐and time‐dependent manner in some ranges . After adding TrkB/Fc ,the levels of intracellular CD11b and BDNF expression and supernatant TNF‐αlevel were decreased with a dose‐and time‐dependent manner in some range .CD11b and BDNF expression levels was decreased in a dose‐and time‐dependent manner .Adding exogenous BDNF ,the levels of intracellular CD11b and BDNF expression and supernatant TNF‐α level were in‐creased again .Conclusion Intracellular BDNF expression is increased when BV2 microglia is activated and replenishing exogenous BDNF can activate microglia .Therefore BDNF may play an important role in the microglia activation process .