Alzheimer's disease (AD) is a neurodegenerative disorder characterized by β-amyloid (Aβ) and tau protein deposition. Microglia are important immune cells in the brain. In the early stage of AD, they play a protective role by clearing AB deposits, but in the late stage, they may aggravate the pathological manifestations of AD through ways such as enhancing neuroinflammation. Triggering receptor expressed on myeloid cell 2 (TREM2), as a core immunomodulatory factor of microglia, maintains the metabolic homeostasis in the brain by bidirectionally regulating Aβ plaque clearance and neuroinflammatory responses through activating downstream signaling pathways. This article reviews the recent advance in structure and function of TREM2 and its role in the pathology and targeted therapy of AD, in order to provide new thoughts for AD treatment targeting TREM2.

Objective: To construct a Family with sequence similarity 50 member A(FAM50A) gene knockout mouse insulinoma pancreatic β-cell line Beta-TC-6 using CRISPR/Cas9 gene editing technology and to prepare polyclonal antibodies specifically recognizing FAM50A. Methods: Two guide RNAs(sgRNAs) targeting the FAM50A gene were designed,and a recombinant plasmid expressing blue fluorescent protein(BFP) was constructed for gene knockout.The successfully constructed plasmid was transfected into Beta-TC-6 cells,and BFP-positive single cells were isolated for clonal expansion.The expanded monoclonal cell lines were genotyped by Sanger sequencing,and FAM50A protein expression was assessed by Western blot.Purified human recombinant FAM50A protein was used to immunize New Zealand rabbits for the preparation of a polyclonal antibody.The specificity of the prepared antibody was then validated using the successfully established FAM50A knockout cell line. Results: A monoclonal cell line with a successful knockout of the FAM50A gene was identified.Sanger sequencing confirmed base deletions at the target site.Western blot analysis showed a complete absence of FAM50A protein expression in this cell line.The prepared polyclonal antibody successfully recognized endogenous murine FAM50A protein in wild-type Beta-TC-6 cells and in hTERT-RPE1 cells overexpressing human FAM50A-GFP fusion protein,while no signal was detected in the FAM50A knockout cells. Conclusion This study successfully established a FAM50A gene knockout Beta-TC-6 cell model and generated a FAM50A polyclonal antibody,providing powerful tools for future research.

Objective To explore the efficacy and action mechanism of a novel phenothiazine derivative,DA414,in rats with intracerebral hemorrhage(ICH).Methods Male Sprague-Dawley(SD)rats(aged 8～10 weeks,weighing 250～300 g)were randomly divided into sham operation,model,and low and high dose DA414 groups[5 and 10 mg/(kg·d)],with 20 animals per group.The size and extent of injury in the ICH area were measured by magnetic resonance imaging(MRI)and histopathological slices.Neurological function was evaluated with a behavioral grading system.Western blotting was used to detect the expression of ferroptosis-related factor,glutathione peroxidase 4(GPX4).Multiplex immunohistochemistry was employed to quantitatively evaluate microglial activation in perihematomal tissue.RT-qPCR was applied to measure the mRNA expression levels of NLRP3 inflammasome components(NLRP3,Caspase-1,IL-1β),pro-inflammatory(IL-18,TLR4,IL-6,TNF-α)and anti-inflammatory cytokines(IL-4,IL-10).The integrity of the blood-brain barrier(BBB)was assessed by Evans blue staining,and the biosafety of DA414 for the liver,kidneys and heart was assessed by HE staining and clinical biochemical tests.Results DA414 significantly promoted the absorption of hematoma,reduced neuronal injury,and improved neurological function scores.DA414 significantly up-regulated the ferroptosis regulatory factor GPX4(P<0.01),and also significantly inhibited the activation of microglia after ICH(P<0.001).RT-qPCR indicated that DA414 treatment resulted in down-regulation of mRNA expression in the inflammasome pathway(NLRP3,Caspase-1,IL-1β,all P<0.01)and pro-inflammatory cytokines(TLR4,IL-6,both P<0.05;IL-18,TNF-α,both P<0.01)and up-regulation of anti-inflammatory cytokines(IL-4,IL-10,both P<0.05),suggesting that DA414 exerts its neuroprotective effect probably by regulating ferroptosis and inflammation.Safety assessment revealed that DA414 had no significant effect on BBB integrity or damage to the liver,kidneys,and heart in rats.Conclusion DA414 exerts significant neuroprotective effects in ICH model by targeted inhibition for ferroptosis and modulating inflammatory response.Our study provides an experimental foundation for ICH treatment.

Objective To develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature,enabling thrombosis modeling and thrombolytic efficacy validation.Methods A tri-level microvascular network chip(300/200/100 μm diameters)with bifurcated architecture was fabricated using soft lithography.Human retinal microvascular endothelial cells(HRMECs)were perfused into channels,with endothelial coverage monitored via phase-contrast microscopy and F-actin staining.Cellular bioactivity was assessed using mitochondrial membrane potential probes(5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide,JC-1)and nitric oxide(NO)quantification.Fresh blood samples from 10 healthy donors(Yongchuan Hospital Affiliated to Chongqing Medical University,March to June 2024)were perfused with digital injection pump to mimic blood flow in human body into 3 experimental groups:normal whole blood,and TNF-α-activated endothelium+normal blood,TNF-α-activated endothelium+TNF-α-treated blood.Three inlet blood flow rates of 37.8、11.1 and 3.5 μL/min were set in each group.Two experimental groups,normal saline and recombinant human tissue-type plasminogen activator(rtPA),were established using the endothelialized microfluidic thrombosis model to validate thrombolytic efficacy.Endothelial functional impacts were assessed through integrated DAPI/NO staining and thrombosis model analysis across 3 intervention phases:pre-thrombosis,post-thrombosis,and post-thrombolysis.Results A tri-level microfluidic vascular model(300/200/100 μm diameters)was successfully constructed.In 72 h after endothelial cell perfusion,complete channel coverage was achieved,with phase-contrast microscopy and F-actin staining confirming confluent cellular alignment.JC-1/NO assays validated preserved endothelial bioactivity.Compared with the whole blood group,both TNF-α-activated endothelium+normal blood and TNF-α-activated endothelium+TNF-α-treated blood groups exhibited significantly increased thrombus occupancy rates at identical flow rates(all P<0.001).Notably,TNF-α-activated endothelium+TNF-α-treated blood group demonstrated the highest thrombus ratio at 3.5 μL/min(P<0.001).The rtPA group showed superior thrombolytic efficacy versus saline(P<0.001).Endothelial monolayer integrity was maintained across intervention phases,with thrombosis triggering significant NO elevation(P<0.001).Conclusion Our retinal vasculature-mimetic microfluidic model enables precise thrombosis modeling and drug evaluation,providing new methodology for studying retinal vascular occlusive diseases.

Helicobacter pylori(H.pylori)infection triggers gastric mucosal inflammatory responses and spasmolytic polypeptide-expressing metaplasia(SPEM).These pathological conditions can escalate the severity of chronic gas-tritis,gastric ulcers and even cause gastric cancer.SPEM is frequently viewed as an early sign of gastric mucosal injury and the onset of carcinogenesis.A comprehensive analysis of the genesis and molecular regulation of SPEM cells in the context of H.pylori infection further has enlightened the pathogenesis of gastric mucosal diseases and provide new ideas and targets for diagnosing and treatment of H.pylori-related gastric mucosal diseases.This paper reviews a variety of molecular biomarkers associated with SPEM,encompassing TFF2,CD44v9,and AQP5,and delineates their pivotal regulatory functions in H.pylori infection and SPEM.This paper also reviews the origination of SPEM cells and pertinent molecular regulatory mechanisms.

Objective:To analyze and compare differences in cortical functioning between patients with post-stroke dysphagia (PSD) following a left hemisphere stroke and healthy individuals using functional near-infrared spectroscopy (fNIRS).Methods:Twenty-six patients recovering from post-stroke dysphagia following a left hemisphere stroke formed the study′s PSD group, and 26 age-matched healthy subjects serves as the HC group. A 41-channel infrared spectroscope was used to record any changes in oxyhemoglobin (HbO) concentration while swallowing and at rest. The fNIRS data were statistically analyzed using Nirspark software. The β-values, reflecting the level of cortical activation, and the swallowing-related specific functional connectivity (FC) strength values (ΔFCs), representing task-specific FC strength, were extracted. The β-values and ΔFCs of the two groups were compared.Results:Compared with the HC group, the PSD group showed significantly reduced activation in Brodmann area (BA) 3/4/6/43 and BA4/6 of the left hemisphere during swallowing. Those areas correspond to the left primary motor cortex (M1), primary somatosensory cortex (S1), premotor cortex, and supplementary motor area (PM). Significantly reduced activation was observed in the PSD group in the right hemisphere at BA45/46/47, BA45/38/48, and BA10, corresponding to the right prefrontal cortex (PFC). The ΔFC values between the left PM-left M1, left PM-left S1, left M1-right S1, and left S1-right M1 in the PSD group were significantly lower than those in the HC group.Conclusions:Left hemispheric PSD is associated not only with decreased activation in the ipsilesional sensorimotor cortex (M1, S1, PM) but also with functional decline in the contra-lesional PFC. During swallowing, persons with left hemispheric PSD exhibit extensive impairment in inter-cortical network connectivity, with particularly marked reductions in connectivity between their ipsilesional and contra-lesional sensorimotor cortices.

Myxedema coma is a rare condition, typically arising from long-standing, untreated hypothyroidism and triggered by factors such as infection, hypothermia, or severe illness. This report details a successfully treated case of myxedema coma with cardiac attest, accompanied by a literature review, to enhance clinical awareness and improve the diagnosis and management of this critical condition.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Lipoatrophic diabetes(LD) is a rare and distinct form of diabetes characterized by notable clinical heterogeneity. It is often considered one of the manifestations of lipodystrophy syndrome(LDS). In clinical practice, LD is frequently misdiagnosed as type 2 diabetes; however, its management protocols and prognostic outcomes differ significantly from those of other diabetes subtypes. Therefore, timely and accurate diagnosis is of great clinical importance. This paper presents two detailed case reports of female patients with LD. Through an in-depth analysis of their clinical features, it also provides an comprehensive review of the key clinical manifestations of LDS, potential pathogenic mechanisms, and current approaches to genetic diagnosis. The aim is to enhance clinicans′ awareness of LDS and improve corresponding diagnostic and therapeutic strategies.

Thyroid eye disease(TED), also known as thyroid-associated ophthalmopathy(TAO) or Graves' ophthalmopathy(GO), is an organ-specific autoimmune disorder associated with autoimmune thyroid disease and can severely impair vision and quality of life. The pathogenesis of TED is closely linked to dysregulated cytokine networks; however, current therapies yield limited response rates in patients with moderate-to-severe TED. This review synthesizes the mechanisms of action of key cytokines and evaluates the translational potential of emerging targeted therapies, aiming to provide a rationale for overcoming current therapeutic bottlenecks and advancing precision treatment strategies.