Structural and functional alterations in the brains of patients with anisometropic and strabismic amblyopia:a systematic review of magnetic resonance imaging studies

Amblyopia is the most common cause of vision loss in children and can persist into adulthood in the absence of effective intervention.Previous clinical and neuroimaging studies have suggested that the neural mechanisms underlying strabismic amblyopia and anisometropic amblyopia may be different.Therefore,we performed a systematic review of magnetic resonance imaging studies investigating brain alterations in patients with these two subtypes of amblyopia;this study is registered with PROSPERO(registration ID:CRD42022349191).We searched three online databases(PubMed,EMBASE,and Web of Science) from inception to April 1,2022;39 studies with 633 patients(324patients with anisometropic amblyo pia and 309 patients with strabismic amblyopia) and 580 healthy controls met the inclusion criteria(e.g.,case-control designed,pee r-reviewed articles) and were included in this review.These studies highlighted that both strabismic amblyopia and anisometropic amblyopia patients showed reduced activation and distorted topological cortical activated maps in the striate and extrastriate co rtices during tas k-based functional magnetic resonance imaging with spatial-frequency stimulus and retinotopic representations,respectively;these may have arisen from abnormal visual experiences.Compensations for amblyopia that are reflected in enhanced spontaneous brain function have been reported in the early visual cortices in the resting state,as well as reduced functional connectivity in the dorsal pathway and structural connections in the ventral pathway in both anisometro pic amblyopia and strabismic amblyopia patients.The shared dysfunction of anisometro pic amblyopia and strabismic amblyopia patients,relative to controls,is also chara cterized by reduced spontaneous brain activity in the oculomotor co rtex,mainly involving the frontal and parietal eye fields and the cerebellu m;this may underlie the neural mechanisms of fixation instability and anomalous saccades in amblyopia.With regards to specific alterations of the two forms of amblyo pia,anisometropic amblyo pia patients suffer more microstructural impairments in the precortical pathway than strabismic amblyopia patients,as reflected by diffusion tensor imaging,and more significant dysfunction and structural loss in the ventral pathway.Strabismic amblyopia patients experience more attenuation of activation in the extrastriate co rtex than in the striate cortex when compared to anisometropic amblyopia patients.Finally,brain structural magnetic resonance imaging alterations tend to be lateralized in the adult anisometropic amblyopia patients,and the patterns of brain alterations are more limited in amblyopic adults than in childre n.In conclusion,magnetic resonance imaging studies provide important insights into the brain alterations underlying the pathophysiology of amblyopia and demonstrate common and specific alte rations in anisometropic amblyo pia and strabismic amblyopia patients;these alterations may improve our understanding of the neural mechanisms underlying amblyopia.

P53单克隆抗体的制备及其临床应用(英文)

Objective: The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initially in clinical immunoassay. Methods: Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by affinity chromatography. Antibody titer was determined by enzyme linked immunosorbent assay(ELISA). The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results: Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by affinity chromatography. The purity of antibodies was higher than90%. The titers of antibodies were more than 1: 6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor cell lines and determine the expression of P53 in tumor tissue. Conclusion: Three strains of monoclonal antibodies with high affinity to P53 were successfully established, which can be used for detecting the expression of P53 in tumor cells or tissue.

Anti-tumor activities of macromolecular fractions of fresh gecko vivo and their induction of Bel-7402 cell differentiation

Objective:To investigate the anti-tumor effect of macromolecular fractions of fresh gecko (M-AG) in vivo and their differentiation-inducing activity in Bel-7402 cells in vitro.Methods:An H22 hepatocarcinoma-bearing mouse model was used to evaluate the anti-tumor activity of M-AG samples.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was applied to analyze cell viability.Cell morphology was observed by phase contrast microscopy.The quantity of the alpha-fetoprotein was detected by a radioimmunoassay.Chromatometry was used to assay the albumin quantity.Activities of alkaline phosphatase and γ-glutamyl trans-peptidase were measured by biochemical methods.Finally,western blotting was applied to assess proteins in the mitogen-activated protein kinase (MAPK) signaling pathway.Results:Macromolecular fractions of fresh gecko exerted a significant anti-tumor effect in mice.The inhibition rate of tumor growth was 63% in the moderate M-AG dose group.Cells treated with M-AG displayed a differentiated state.The treatment lowered alphafetoprotein secretion and significantly decreased the activities of γ-glutamyl trans-peptidase and alkaline phosphatase in Bel-7402 cells.In contrast,M-AG increased the amount of albumin in the cell culture medium.All biochemical indices demonstrated that M-AG induced Bel7402 cell differentiation.Western blotting showed no changes in the quantities of extracellular signal-regulated kinase (ERK) 1/2,p38MAPK,or c-Jun N-terminal protein kinase 1/2.However,M-AG significantly activated the phosphorylation of ERK1/2 in a dose-dependent manner.In addition,M-AG had no significant influence on the expression of nuclear factor-kappa B.Conclusion:Macromolecular fractions of fresh gecko has an anti-tumor activity in H22 hepatocarcinoma-bearing mice in vivo and inhibits Bel-7402 cell proliferation in vitro by inducing cell differentiation related to activation of ERK1/2.

二甲双胍抑制人肝癌细胞HepG2的实验研究(英文)

Objective: This work aimed to study the inhibitory effect and the related mechanism of metformin(MET) on the proliferation of human hepatoma HepG2 cells. Methods: Human hepatoma HepG2 cells were treated with MET(0, 2, 10, and 50 mM). The inhibitory effect of MET on the proliferation of HepG2 cells was determined by MTT method. The apoptosis of HepG2 cells was detected by flow cytometry. The expression of cyclin D1 in HepG2 cells was examined by Western blot. ROS-DHE fluorescence probe was used to stain the reactive oxygen species(ROS) generated by HepG2 cells after treatment. Results: MET could inhibit the proliferation of HepG2 cells in a dose and time dependent manner. MET promoted the apoptosis of HepG2 cells. In addition, MET suppressed the expression of cell cycle protein cyclin D1 and induced the production of ROS in HepG2 cells. Conclusion: MET can inhibit the proliferation of human hepatoma HepG2 cells and induce cell apoptosis. Meanwhile, MET has the ability to decrease the expression of cyclin D1 and induce ROS generation, which may be involved in the mechanism of inhibiting hepatoma cells proliferation.

Visfatin and 25-Hydroxyvitamin D_(3) Levels Affect Coronary Collateral Circulation Development in Patients with Chronic Coronary Total Occlusion

Background:Coronary collateral circulation(CCC)plays a vital role in the myocardial blood supply,especially for ischemic myocardium.Evidence suggests that the visfatin and 25-hydroxyvitamin D_(3)[25(OH)D_(3)]levels are related to the degree and incidence of vascular stenosis associated with coronary artery disease;however,few studies have evaluated the effect of visfatin and 25(OH)D_(3) on CCC development in patients with chronic total occlusion(CTO).This study aimed to evaluate the relationship between the serum visfatin and 25(OH)D_(3) levels and CCC in patients with CTO.Methods:A total of 189 patients with CTO confirmed by coronary angiography were included.CCC was graded from 0 to 3 according to the Rentrop-Cohen classification.Patients with grade 0 or grade 1 collateral development were in-cluded in the poor CCC group(n=82),whereas patients with grade 2 or grade 3 collateral development were included in the good CCC group(n=107).The serum visfatin and 25(OH)D_(3) levels were measured by ELISA.Results:The visfatin level was significantly higher in the poor CCC group than in the good CCC group,and the 25(OH)D_(3) level was significantly lower in the poor CCC group than in the good CCC group(P=0.000).Correlation analysis showed that the Rentrop grade was negatively correlated with the visfatin level(r=−0.692,P=0.000)but positively correlated with the 25(OH)D_(3) level(r=0.635,P=0.000).Logistic regression analysis showed that the vis-fatin and 25(OH)D_(3) levels were independent risk factors for CCC(odds ratio 1.597,95%confidence interval 1.300-1.961,P=0.000 and odds ratio 0.566,95%confidence interval 0.444-0.722,P=0.000,respectively).The visfatin and 25(OH)D_(3) levels can effectively predict the CCC status.Conclusion:Serum visfatin and 25(OH)D_(3) levels are related to CCC development and are independent predictors of poor CCC.

Cryo-EM structure of cannabinoid receptor CB1-β-arrestincomplex

Dear Editor,G protein-coupled receptors(GPCRs)play a vital role in regulating almost every aspect of human physiology,making up more than one-third of marketed drug targets(Santos et al.,2017).GPCRs orchestrate their signalling through interactions with three distinct downstream protein families:G proteins,G protein-coupled receptor kinases(GRKs),and arrestins(Santos et al.,2017).While G protein-mediated signalling is initiated upon GPCR stimulation,activated GPCRs return to their basal levels through a GRK-and arrestin-regulated desensitization process(Santos et al.,2017).In addition to modulating receptor desensitization,β-arrestin also regulates downstream events that are distinct from classical G protein signalling(Ahn et al.,2020).

Design of recognition algorithm for multiclass digital display instrument based on convolution neural network

Digital display instrument identification is a crucial approach for automating the collection of digital display data.In this study,we propose a digital display area detection CTPNpro algorithm to address the problem of recognizing multiclass digital display instruments.We developed a multiclass digital display instrument recognition algorithm by combining the character recognition network constructed using a convolutional neural network and bidirectional variable-length long short-term memory(LSTM).First,the digital display region detection CTPNpro network framework was designed based on the CTPN network architecture by introducing feature fusion and residual structure.Next,the digital display instrument identification network was constructed based on a convolutional neural network using twoway LSTM and Connectionist temporal classification(CTC)of indefinite length.Finally,an automatic calibration system for digital display instruments was built,and a multiclass digital display instrument dataset was constructed by sampling in the system.We compared the performance of the CTPNpro algorithm with other methods using this dataset to validate the effectiveness and robustness of the proposed algorithm.

Tet2调节骨髓间充质干细胞功能

Tet2(Tet家族成员2)在DNA去甲基化修饰、表观遗传调控及骨髓造血中起着重要作用。笔者课题组前期研究发现,随着年龄增长,Tet2敲除小鼠逐步发展为淋系白血病和髓系白血病。但Tet2在骨髓微环境中的作用仍不清楚。进一步研究发现,Tet2敲除的骨髓间充质干细胞(Mesenchymal stem cells,MSC)更多处于G2/M分裂期,其细胞分裂时间缩短,生长速度加快。长周期培养-起始细胞实验表明,Tet2敲除的MSC支持造血干细胞扩增和髓系分化的能力增强。通过点杂交实验发现,Tet2敲除后,骨髓细胞DNA总甲基化水平升高。对Tet2缺失的骨髓细胞进行甲基化测序,结果表明:基因组转录调控区域等多个功能性结构域的甲基化水平明显升高。同时,敲除Tet2的MSC分泌IL-8、IL-18等炎性细胞因子的能力下降;敲除Tet2的MSC更多分泌促进造血干细胞髓系分化的GM-CSF和CCL-3等细胞因子。Tet2可以影响间充质干细胞造血支持作用,进而调节造血。

Arabidopsis ANAC092 regulates auxin-mediated root development by binding to the ARF8 and PIN4 promoters

Auxin is an important plant hormone that is essential for growth and development due to its effects on organogenesis, morphogenesis, tropisms, and apical dominance. The functional diversity of auxin highlights the importance of its biosynthesis, transport, and associated responses. In this study, we show that a NAC transcription factor, ANAC092(also named At NAC2 and ORESARA1),known to positively regulate leaf senescence and contribute to abiotic stress responses, also affects primary root development. Plants overexpressing ANAC092 had altered root meristem lengths and shorter primary roots compared with the wild-type control. Additionally, expression of the pro ANAC092::GUS was strongly induced by indole-3-acetic acid. Quantitative real-time RT-PCR(q RT-PCR) analysis revealed that the YUCCA2, PIN, and ARF expression levels were downregulated in ANAC092-overexpressing plants. Moreover, yeast one-hybrid and chromatin immunoprecipitation assays confirmed that ANAC092 binds to the promoters of AUXIN RESPONSE FACTOR 8(ARF8) and PIN-FORMED 4(PIN4). Furthermore, a dual-luciferase assay indicated that ANAC092 decreases ARF8 and PIN4 promoter activities. We also applied a CRISPR/Cas9 system to mutate ANAC092. The roots of three of the analyzed mutants were longer than normal. Collectively, our findings indicate that ANAC092 negatively affects root development by controlling the auxin pathway.

Novel small molecule retrograde transport blocker confers post-exposure protection against ricin intoxication

Ricin is a highly toxic type 2 ribosome-inactivating protein(RIP)which is extracted from the seeds of castor beans.Ricin is considered a potential bioterror agent and no effective antidote for ricin exists so far.In this study,by structural modification of a retrograde transport blocker Retro-2cyc1,a series of novel compounds were obtained.The primary screen revealed that compound 27 has an improved antiricin activity compare to positive control.In vitro pre-exposure evaluation in Madin-Darby Canine Kidney(MDCK)cells demonstrated that 27 is a powerful anti-ricin compound with an EC50 of 41.05 nmol/L against one LC(lethal concentration,5.56 ng/mL)of ricin.Further studies surprisingly indicated that 27 confers post-exposure activity against ricin intoxication.An in vivo study showed that 1 h post-exposure administration of 27 can improve the survival rate as well as delay the death of ricin-intoxicated mice.A drug combination of 27 with monoclonal antibody mAb4 C13 rescued mice from one LD(lethal dose)ricin challenge and the survival rate of tested animals is 100%.These results represent,for the first time,indication that small molecule retrograde transport blocker confers both in vitro and in vivo post-exposure protection against ricin and therefore provides a promising candidate for the development of anti-ricin medicines.

Assessment of Spinal Tumor Treatment Using Implanted 3D-Printed Vertebral Bodies with Robotic Stereotactic Radiotherapy

To investigate the feasibility and early efficacy of 3D-printed vertebral body implantation combined with robotic radiosurgery in the treatment of spinal tumors.This study included 14 patients with spinal tumors from December 2017 to June 2018.Before surgery,all patients were subjected to CT scan and 3D data of the corresponding vertebral segments were collected.Titanium alloy formed 3D-printed vertebral body implantation and robotic stereotactic radiotherapy were performed because of the risk of postoperative residual,high risk of recovery,or recurrence after surgery.The main outcomes included the remission of symptoms,vertebral body stability,robotic stereotactic surgical precision,and local tumor control.All patients received complete and successful combination therapy,and all healed primarily without complications.The error of the coverage accuracy for robotic radiosurgery was less than 0.5 mm,and the error of the rotation angle was less than 0.5
.The therapeutic toxicity was limited(mainly in grades 1–2),and adverse events were uncommon.The evaluation of vertebral body stability and histocompatibility for all patients met the postoperative clinical requirements.For patients with post spinal injury,the pain symptoms were reduced or disappeared(93%),and nerve function was improved or even recovered after treatment(100%).During our follow-up period,most tumors were locally well controlled(93%).3D-printed vertebral body implantation combined with robotic radiosurgery may offer a new treatment of spinal tumors.Chinese clinical trial registry:ChiCTR-ONN-17013946.

NIR-activated nanosystems with self-modulated bacteria targeting for enhanced biofilm eradication and caries prevention

The efficacious delivery of antimicrobial drugs to intractable oral biofilms remains a challenge due to inadequate biofilm penetration and lack of pathogen targeting.Herein,we have developed a microenvironment-activated poly(ethylene glycol)(PEG)-sheddable nanoplatform to mediate targeted delivery of drugs into oral biofilms for the efficient prevention of dental caries.The PEGylated nanoplatform with enhanced biofilm penetration is capable of deshielding the PEG layer under slightly acidic conditions in a PEG chain length-dependent manner to re-expose the bacteria-targeting ligands,thereby facilitating targeted codelivery of ciprofloxacin(CIP)and IR780 to the bacteria after accumulation within biofilms.The nanoplatform tends to induce bacterial agglomeration and suffers from degradation in the acidic oral biofilm microenvironment,triggering rapid drug release on demand around bacterial cells.The self-modulating nanoplatform under near-infrared(NIR)irradiation accordingly displays greatly augmented potency in oral biofilm penetration and disruption compared with drugs alone.Topical oral treatment with nanoplatforms involving synergetic pharmacological and photothermal/photodynamic trinary therapy results in robust biofilm dispersion and efficacious suppression of severe tooth decay in rats.This versatile nanoplatform can promote local accumulation and specific drug transport into biofilms and represents a new paradigm for targeted drug delivery for the management of oral biofilm-associated infections.

Characteristics of patients with chronic airflow obstruction caused by solid fuel or tobacco smoke

To the Editor: Chronic airflow obstruction (CAO) is a characteristic feature of chronic obstructive pulmonary disease (COPD) and occurs due to airway and/or alveolar abnormalities typically associated with exposure to noxious particles or gases.[1] The major risk factor for CAO is cigarette smoking, but exposure to solid fuel likely influences CAO development. Studies have found that solid fuel exposure is associated with a high prevalence of CAO, particularly among women.[2] Comparing COPD caused by either solid fuel or tobacco smoke exposure is very significant because about 3 billion people are exposed to solid fuel smoke, and 1.01 billion people smoke tobacco, globally. This study aimed to investigate and compare the clinical and functional characteristics of CAO patients exposed to solid fuel and tobacco smoke using propensity score matching (PSM) in western China.

DNAH2 facilitates the homologous recombination repair of Fanconi anemia pathway through modulating FANCD2 ubiquitination

Fanconi anemia(FA),an X-linked genetic or autosomal recessive disease,exhibits complicated pathogenesis.Previously,we detected the mutated Dynein Axonemal Heavy Chain 2(DNAH2)gene in 2 FA cases.Herein,we further investigated the potential association between DNAH2 and the homologous recombination repair pathway of FA.The assays of homologous recombination repair,mitomycin C(MMC)sensitivity,immunofluorescence,and ubiquitination modification were performed in U2OS and DR-U2OS cell lines.In MMC-treated U2OS cells,the downregulation of the DNAH2 gene increased the sensitivity of cells to DNA inter-strand crosslinks.We also observed the reduced enrichment of FANCD2 protein to DNA damage sites.Furthermore,the ubiquitination modification level of FANCD2 was influenced by the deficiency of DNAH2.Thus,our results suggest that DNAH2 may modulate the cell homologous recombination repair partially by increasing the ubiquitination and the enrichment to DNA damage sites of FANCD2.DNAH2 may act as a novel co-pathogenic gene of FA patients.