Alzheimer's disease (AD), a progressive dementia, is one of the most common neurodegenerative diseases. Clinical trial results of amyloid-β (Aβ) and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing. There are currently no effective strategies for slowing the progression of AD. Herein, we spotlight the dysregulation of lipid metabolism, particularly the elevation of ceramides (Cers), as a critical yet underexplored facet of AD pathogenesis. Our study delineates the role of Cers in promoting microglial pyroptosis, a form of programmed cell death distinct from apoptosis and necroptosis, characterized by cellular swelling, and membrane rupture mediated by the NLRP3 inflammasome pathway. Utilizing both in vivo experiments with amyloid precursor protein (APP)/presenilin 1 (PS1) transgenic mice and in vitro assays with BV-2 microglial cells, we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin (ICA), a flavonoid with known antioxidant and anti-inflammatory properties. Our findings reveal a significant increase in Cers levels and pyroptosis markers (NOD-like receptor family, pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain, caspase-1, gasdermin D (gasdermin D (GSDMD)), and interleukin-18 (IL-18)) in the brains of AD model mice, indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis. Conversely, ICA treatment effectively reduces these pyroptotic markers and Cer levels, thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD. This study not only advances our understanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy, capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2 (COX-2)-NLRP3 inflammasome-gasdermin D (GSDMD) axis. Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of targeting microglial pyroptosis in AD.

Objective:To investigate the role of human dermal microvascular endothelial cells(HMEC-1)exosome hsa-miR-4488_L in regulating the osteogenic and lipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)and to elucidate the mechanism of action underlying fate differentiation.Methods:The hsa-miR-4488_L mimic or negative control mimic was transfected into BMSCs, which were then cultured under osteogenic or lipogenic conditions, respectively.RT-qPCR was employed to detect the mRNA expression levels of osteogenic and lipogenic genes in BMSCs.Alizarin red staining was utilized to assess the osteogenic differentiation capability of hsa-miR-4488_L in BMSCs, while oil red O staining was used to evaluate the lipogenic differentiation potential of BMSCs.The mimic control and hsa-miR-4488_L mimic were transfected into elderly BMSCs, and age-related phenotypes were assessed using RT-qPCR and SA-β-gal staining.The direct target genes of hsa-miR-4488_L acting on BMSCs were identified through bioinformatics analysis and subsequently validated by RT-qPCR and Western blot.Results:After treatment with the hsa-miR-4488_L mimic, the expressions of osteogenic-related genes ALP( P=0.007), BSP( P=0.001), and Col1( P<0.001)in BMSCs were upregulated.Additionally, alizarin red staining results indicated an increase in the number of calcified nodules Pparγ( P=0.002).Concurrently, under adipogenic induction conditions, the adipogenic-related genes Pparγ( P=0.008)and Perilipin( P<0.001)were significantly downregulated in the hsa-miR-4488_L mimic treatment group, and oil red O staining demonstrated a reduction in lipid droplet production( P=0.032).The mRNA expression of the aging-related gene P16( P=0.009)was downregulated following treatment with the hsa-miR-4488_L mimic, and the number of senescent cells decreased as indicated by SA-β-gal staining.Bioinformatics analysis revealed that Smad3 was the direct target gene of hsa-miR-4488_L in BMSCs.RT-qPCR results confirmed that the expression of Smad3 was downregulated after treatment with the hsa-miR-4488_L mimic( P=0.040).Furthermore, Western blot analysis showed a reduction in the protein level of Smad3. Conclusions:HMEC-EXOs-derived hsa-miR-4488_L regulates the balance between osteogenic and adipogenic differentiation in BMSCs through Smad3.Consequently, hsa-miR-4488_L may serve as a potential miRNA biomarker for age-related osteoporosis.

Objective:To develop a convolutional neural network model of whole slide imaging of cerebrospinal fluid cells for rapid and accurate identification and classification of tumor cells in cerebrospinal fluid.Methods:A total of 8 692 cerebrospinal fluid cytology smears from Huashan Hospital Affiliated to Fudan University from January 2nd, 2019, to December 27th, 2024. As randomly assigned, the training set included 4 941 benign and 1 745 malignant samples, while the validation set comprised of 1 368 benign and 638 malignant samples. Whole-slide digital images were acquired using a cytopathology scanner, cells (clusters) were annotated for classification, and a deep learning model was constructed via tiled image patches for cell detection and classification. Model performance was evaluated using accuracy, sensitivity, specificity, and other indicators. The classification efficiency of manual microscopy was compared.Results:The model achieved a mean precision of 96.75% for cerebrospinal fluid cell classification. For malignant tumor cells, the classification accuracy was 96.61% (mAP=98.36%, AUC=0.97). Subtype classification accuracies for epithelial/epithelioid tumors and small round cell tumors were 97.13% (AUC=0.98) and 95.58% (AUC=0.93), respectively. Compared with manual microscopy, which took (9.70±0.82) minutes for classifying 200 cells, (18.27±1.21) minutes for 500 cells, and often exceeded 60 minutes or infeasible for full slides, the AI model took (3.46±0.49) seconds for 200 cells, (6.76±0.82) seconds for 500 cells, and a median of 48.57 seconds for full slides ( P<0.001), representing an efficiency improvement of approximately 161-170 times, significantly enhancing diagnostic efficiency. Conclusion:This fully automated hierarchical deep learning model enables efficient and accurate tumor cell identification and classification in CSF, providing an effective auxiliary tool for the rapid diagnosis of meningeal carcinomatosis.

Alzheimer's disease(AD),a progressive dementia,is one of the most common neurodegenerative dis-eases.Clinical trial results of amyloid-β(Aβ)and tau regulators based on the pretext of straightforward amyloid and tau immunotherapy were disappointing.There are currently no effective strategies for slowing the progression of AD.Herein,we spotlight the dysregulation of lipid metabolism,particularly the elevation of ceramides(Cers),as a critical yet underexplored facet of AD pathogenesis.Our study delineates the role of Cers in promoting microglial pyroptosis,a form of programmed cell death distinct from apoptosis and necroptosis,characterized by cellular swelling,and membrane rupture mediated by the NLRP3 inflammasome pathway.Utilizing both in vivo experiments with amyloid precursor protein(APP)/presenilin 1(PS1)transgenic mice and in vitro assays with BV-2 microglial cells,we investigate the activation of microglial pyroptosis by Cers and its inhibition by icariin(ICA),a flavonoid with known antioxidant and anti-inflammatory properties.Our findings reveal a significant increase in Cers levels and pyroptosis markers(NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain,caspase-1,gasdermin D(GSDMD),and interleukin-18(IL-18))in the brains of AD model mice,indicating a direct involvement of Cers in AD pathology through the induction of microglial pyroptosis.Conversely,ICA treatment effec-tively reduces these pyroptotic markers and Cer levels,thereby attenuating microglial pyroptosis and suggesting a novel therapeutic mechanism of action against AD.This study not only advances our un-derstanding of the pathogenic role of Cers in AD but also introduces ICA as a promising candidate for AD therapy,capable of mitigating neuroinflammation and pyroptosis through the cyclooxygenase-2(COX-2)-NLRP3 inflammasome-gasdermin D(GSDMD)axis.Our results pave the way for further exploration of Cer metabolism disorders in neurodegenerative diseases and highlight the therapeutic potential of tar-geting microglial pyroptosis in AD.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Objective To summarise the best evidence on non-pharmacological management in children and to provide evidence-based guidelines for clinical practice.Methods With the 6S evidence pyramid model,a comprehensive and systematic search across multiple databases was conducted,including UpToDate,BMJ Best Practice,Joanna Briggs Institute of Australia's Centre for Evidence-based Health Care Database(JBI),National Guideline Clearing-house(NGC),Guidelines International Network(GIN),The National Institute for Health and Care Excellence(NICE)website,Scottish Intercollegiate Guidelines Network(SIGN),American Dental Association,Canadian Dental Association,Cochrane Library,CINAHL,Embase,PubMed,SinoMed,CNKI and Wanfang Data.The search focused on literature pertaining to the improvement of non-pharmacological strategies for compliance with paediatric oral treatment,encompassing clinical decisions,evidence summaries,clinical guidelines,systematic reviews,expert consensus,best practices,and randomised controlled trials.The literature search encompassed all available publications from the inception of databases up to 5th November,2023.A quality assessment of the literature was independently conducted by four researchers trained by evidence-based nursing courses,while evidence extraction and summarisation were handled by two researchers.Results A total of 16 papers were included,comprising 2 clinical decisions,2 evidence summaries,3 guidelines,5 systematic evaluations,1 best practice,2 expert consensus and 1 randomised controlled trial.Nineteen pieces of evidence were extracted and classified into six categories:outpatient setting,assessment and management of children,pre-treatment non-pharmacological management,in-treatment non-pharmacological management,post-treatment non-pharmacological management and training and assessment.Conclusion This study summarises the best evidences for non-pharmacological management aiming to improve the oral treatment compliance in children.Healthcare providers can facilitate the translation of this evidence into clinical practice by considering the specific clinical context as well as factors such as the age and psychological characteristics of children.

Objective:To learn about the clinical manifestations, laboratory test results, and prognosis of patients with acute and chronic brucellosis, and to provide a scientific basis for clinical diagnosis and treatment.Methods:A retrospective study was conducted to collect the clinical data of 776 patients with brucellosis admitted to the First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2023. The patients were divided into acute phase group and chronic phase group according to the course of disease. Their clinical manifestations, laboratory test results, and prognosis were compared and analyzed between the two groups.Results:Out of 776 patients with brucellosis, 649 were in the acute phase group and 127 were in the chronic phase group. Male accounted for 71.6% (556/776). The age was (44.91 ± 0.60) years old, mainly concentrated in the age range of 46 - 60 years old (38.0%, 295/776). Farmers were the main occupation, accounting for 58.4% (453/776). The disease occurred in all months of the year, with autumn being the peak period (33.9%, 263/776). The primary clinical manifestations were fever (64.7%, 502/776) and fatigue (40.5%, 314/776). The incidence rates of fever and fatigue in the acute phase group were higher than those in the chronic phase group [70.4% (457/649) vs. 35.4% (45/127), 42.5% (276/649) vs. 29.9% (38/127)], with statistically significant differences ( χ2 = 56.91, 7.01, P < 0.05). Complications of the osteoarticular system were the most common, accounting for 47.8% (371/776). Abnormal results of blood routine examinations were mainly characterized by decreased hemoglobin, decreased lymphocytes, and decreased white blood cells, accounting for 53.7% (417/776), 32.6% (253/776), and 22.6% (175/776), respectively. The incidence rate of decreased hemoglobin in the acute phase group was higher than that in the chronic phase group [63.5% (412/649) vs. 3.9% (5/127)], with a statistically significant difference ( χ2 = 151.49, P < 0.001). The remaining abnormal laboratory test results were mainly characterized by elevated erythrocyte sedimentation rate, elevated C-reactive protein, and elevated aspartate aminotransferase, accounting for 75.1% (583/776), 50.6% (393/776), and 39.8% (309/776), respectively. The incidence rates of elevated erythrocyte sedimentation rate and elevated aspartate aminotransferase in the acute phase group were higher than those in the chronic phase group [77.8% (505/649) vs. 61.4% (78/127), 43.3% (281/649) vs. 22.0% (28/127)], with statistically significant differences ( χ2 = 15.28, 20.02, P < 0.001). The overall positive rate of Brucella blood culture was 57.5% (446/776), and there was a statistically significant difference in blood culture between the two groups ( χ2 = 17.08, P = 0.002). The positive rate of blood culture in the acute phase group was higher than that in the chronic phase group [60.1% (390/649) vs. 44.1% (56/127)]. Ninety-four point three percent (732/776) of patients were treated with antibiotics, with rifampicin + doxycycline as the main treatment regimen (45.2%, 331/732). The median of antibiotic types used in the acute and chronic phase groups were 3 and 4, respectively. The overall incidence rate of adverse drug reactions was 3.8% (28/732). Eighty-seven point five percent (625/714) of patients improved or recovered after treatment, while 12.5% (89/714) did not recover or experienced relapse. Conclusions:The main clinical manifestations of brucellosis patients are fever and fatigue, with a higher incidence of complications in the osteoarticular system, and a better prognosis. The incidence of fever, fatigue, decreased hemoglobin, elevated erythrocyte sedimentation rate, elevated aspartate aminotransferase, and positive Brucella blood culture in patients in the acute phase are higher, and the types of antibiotics used are fewer than those in patients in the chronic phase.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Objective To summarise the best evidence on non-pharmacological management in children and to provide evidence-based guidelines for clinical practice.Methods With the 6S evidence pyramid model,a comprehensive and systematic search across multiple databases was conducted,including UpToDate,BMJ Best Practice,Joanna Briggs Institute of Australia's Centre for Evidence-based Health Care Database(JBI),National Guideline Clearing-house(NGC),Guidelines International Network(GIN),The National Institute for Health and Care Excellence(NICE)website,Scottish Intercollegiate Guidelines Network(SIGN),American Dental Association,Canadian Dental Association,Cochrane Library,CINAHL,Embase,PubMed,SinoMed,CNKI and Wanfang Data.The search focused on literature pertaining to the improvement of non-pharmacological strategies for compliance with paediatric oral treatment,encompassing clinical decisions,evidence summaries,clinical guidelines,systematic reviews,expert consensus,best practices,and randomised controlled trials.The literature search encompassed all available publications from the inception of databases up to 5th November,2023.A quality assessment of the literature was independently conducted by four researchers trained by evidence-based nursing courses,while evidence extraction and summarisation were handled by two researchers.Results A total of 16 papers were included,comprising 2 clinical decisions,2 evidence summaries,3 guidelines,5 systematic evaluations,1 best practice,2 expert consensus and 1 randomised controlled trial.Nineteen pieces of evidence were extracted and classified into six categories:outpatient setting,assessment and management of children,pre-treatment non-pharmacological management,in-treatment non-pharmacological management,post-treatment non-pharmacological management and training and assessment.Conclusion This study summarises the best evidences for non-pharmacological management aiming to improve the oral treatment compliance in children.Healthcare providers can facilitate the translation of this evidence into clinical practice by considering the specific clinical context as well as factors such as the age and psychological characteristics of children.

Objective:To learn about the clinical manifestations, laboratory test results, and prognosis of patients with acute and chronic brucellosis, and to provide a scientific basis for clinical diagnosis and treatment.Methods:A retrospective study was conducted to collect the clinical data of 776 patients with brucellosis admitted to the First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2023. The patients were divided into acute phase group and chronic phase group according to the course of disease. Their clinical manifestations, laboratory test results, and prognosis were compared and analyzed between the two groups.Results:Out of 776 patients with brucellosis, 649 were in the acute phase group and 127 were in the chronic phase group. Male accounted for 71.6% (556/776). The age was (44.91 ± 0.60) years old, mainly concentrated in the age range of 46 - 60 years old (38.0%, 295/776). Farmers were the main occupation, accounting for 58.4% (453/776). The disease occurred in all months of the year, with autumn being the peak period (33.9%, 263/776). The primary clinical manifestations were fever (64.7%, 502/776) and fatigue (40.5%, 314/776). The incidence rates of fever and fatigue in the acute phase group were higher than those in the chronic phase group [70.4% (457/649) vs. 35.4% (45/127), 42.5% (276/649) vs. 29.9% (38/127)], with statistically significant differences ( χ2 = 56.91, 7.01, P < 0.05). Complications of the osteoarticular system were the most common, accounting for 47.8% (371/776). Abnormal results of blood routine examinations were mainly characterized by decreased hemoglobin, decreased lymphocytes, and decreased white blood cells, accounting for 53.7% (417/776), 32.6% (253/776), and 22.6% (175/776), respectively. The incidence rate of decreased hemoglobin in the acute phase group was higher than that in the chronic phase group [63.5% (412/649) vs. 3.9% (5/127)], with a statistically significant difference ( χ2 = 151.49, P < 0.001). The remaining abnormal laboratory test results were mainly characterized by elevated erythrocyte sedimentation rate, elevated C-reactive protein, and elevated aspartate aminotransferase, accounting for 75.1% (583/776), 50.6% (393/776), and 39.8% (309/776), respectively. The incidence rates of elevated erythrocyte sedimentation rate and elevated aspartate aminotransferase in the acute phase group were higher than those in the chronic phase group [77.8% (505/649) vs. 61.4% (78/127), 43.3% (281/649) vs. 22.0% (28/127)], with statistically significant differences ( χ2 = 15.28, 20.02, P < 0.001). The overall positive rate of Brucella blood culture was 57.5% (446/776), and there was a statistically significant difference in blood culture between the two groups ( χ2 = 17.08, P = 0.002). The positive rate of blood culture in the acute phase group was higher than that in the chronic phase group [60.1% (390/649) vs. 44.1% (56/127)]. Ninety-four point three percent (732/776) of patients were treated with antibiotics, with rifampicin + doxycycline as the main treatment regimen (45.2%, 331/732). The median of antibiotic types used in the acute and chronic phase groups were 3 and 4, respectively. The overall incidence rate of adverse drug reactions was 3.8% (28/732). Eighty-seven point five percent (625/714) of patients improved or recovered after treatment, while 12.5% (89/714) did not recover or experienced relapse. Conclusions:The main clinical manifestations of brucellosis patients are fever and fatigue, with a higher incidence of complications in the osteoarticular system, and a better prognosis. The incidence of fever, fatigue, decreased hemoglobin, elevated erythrocyte sedimentation rate, elevated aspartate aminotransferase, and positive Brucella blood culture in patients in the acute phase are higher, and the types of antibiotics used are fewer than those in patients in the chronic phase.