Recent data suggest that vascular endothelial growth factor receptor inhibitor (VEGFRi) can enhance the anti-tumor activity of the anti-programmed cell death-1 (anti-PD-1) antibody in colorectal cancer (CRC) with microsatellite stability (MSS). However, the comparison between this combination and standard third-line VEGFRi treatment is not performed, and reliable biomarkers are still lacking. We retrospectively enrolled MSS CRC patients receiving anti-PD-1 antibody plus VEGFRi (combination group, n=54) or VEGFRi alone (VEGFRi group, n=32), and their efficacy and safety were evaluated. We additionally examined the immune characteristics of the MSS CRC tumor microenvironment (TME) through single-cell and spatial transcriptomic data, and an MSS CRC immune cell-related signature (MCICRS) that can be used to predict the clinical outcomes of MSS CRC patients receiving immunotherapy was developed and validated in our in-house cohort. Compared with VEGFRi alone, the combination of anti-PD-1 antibody and VEGFRi exhibited a prolonged survival benefit (median progression-free survival: 4.4 vs. 2.0 months, P=0.0024; median overall survival: 10.2 vs. 5.2 months, P=0.0038) and a similar adverse event incidence. Through single-cell and spatial transcriptomic analysis, we determined ten MSS CRC-enriched immune cell types and their spatial distribution, including naive CD4⁺ T, regulatory CD4⁺ T, CD4⁺ Th17, exhausted CD8⁺ T, cytotoxic CD8⁺ T, proliferated CD8⁺ T, natural killer (NK) cells, plasma, and classical and intermediate monocytes. Based on a systemic meta-analysis and ten machine learning algorithms, we obtained MCICRS, an independent risk factor for the prognosis of MSS CRC patients. Further analyses demonstrated that the low-MCICRS group presented a higher immune cell infiltration and immune-related pathway activation, and hence a significant relation with the superior efficacy of pan-cancer immunotherapy. More importantly, the predictive value of MCICRS in MSS CRC patients receiving immunotherapy was also validated with an in-house cohort. Anti-PD-1 antibody combined with VEGFRi presented an improved clinical benefit in MSS CRC with manageable toxicity. MCICRS could serve as a robust and promising tool to predict clinical outcomes for individual MSS CRC patients receiving immunotherapy.
Humans ; Colorectal Neoplasms/drug therapy* ; Male ; Female ; Immunotherapy ; Middle Aged ; Aged ; Tumor Microenvironment/immunology* ; Retrospective Studies ; Microsatellite Instability ; Transcriptome ; Single-Cell Analysis ; Programmed Cell Death 1 Receptor/immunology* ; Gene Expression Profiling ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors*

Objective:To explore molecular mechanism of high-altitude hypoxia regulates PPAR signaling pathway induced ferroptosis in spleen.Methods:Hypoxia animal model was constructed,target genes were screened and predicted by combination of transcriptomics and protein omics.Key genes in PPAR and ferroptosis pathways under hypoxia exposure were explored by GO and KEGG enrichment analysis and verified by RT-qPCR and Western blot.Results:Combination of transcriptomics and protein omics showed that 95 predicted target genes(protein)showed significantly differential expression under hypoxic exposure.GO annotation analysis and KEGG enrichment analysis showed that differential genes were mainly significantly enriched in PPAR and ferroptosis signaling pathways.A negative correlation was found between PPAR and ferroptosis signaling pathways,and GSEA showed that differential gene sets of PPAR and ferroptosis signaling pathways exhibited opposite expression trend in high-altitude hypoxia group.Validation of key genes PPARA,RXRB,APOA1 and SCD-1 in PPAR signaling pathway revealed that both mRNA and protein expres-sions were down-regulated under hypoxic exposure.Subsequently,differential expression was observed in mRNA and protein expres-sions of GPX4 in endogenous pathway and SLC7A11,TRP53 and TFRC in exogenous pathway in ferroptosis signaling pathway.Corre-lations between four key genes for ferroptosis and differential inflammation-associated genes(DE-IRGs)were positively or negatively.IL-1β,IL-6,IL-12,IL-18,IFN-γ and TNF-α expressions in spleen tissue were up-regulated under hypoxic exposure.Conclusion:High-altitude hypoxia exposure further induces ferroptosis through PPAR signaling pathway-mediated lipid metabolism disorders,and accompanied by occurrence of inflammatory response,which causes damage of spleen tissue.

BACKGROUND: Generalized pustular psoriasis (GPP), a rare and recurrent autoinflammatory disease, imposes a substantial burden on patients and society. Awareness of GPP in China remains limited. METHODS: This cross-sectional survey, conducted between September 2021 and May 2023 across 14 hospitals in China, included GPP patients of all ages and disease phases. Data collected encompassed demographics, clinical characteristics, economic impact, disease severity, quality of life, and treatment-related complications. Risk factors for GPP recurrence were analyzed. RESULTS: Among 127 patients (female/male ratio = 1.35:1), the mean age of disease onset was 25 years (1st quartile [Q1]-3rd quartile [Q3]: 11-44 years); 29.2% had experienced GPP for more than 10 years. Recurrence occurred in 75.6% of patients, and nearly half reported no identifiable triggers. Younger age at disease onset ( P  = 0.021) and transitioning to plaque psoriasis ( P  = 0.022) were associated with higher recurrence rates. The median diagnostic delay was 8 months (Q1-Q3: 2-41 months), and 32.3% of patients reported misdiagnoses. Comorbidities were present in 53.5% of patients, whereas 51.1% experienced systemic complications during treatment. Depression and anxiety affected 84.5% and 95.6% of patients, respectively. During GPP flares, the median Dermatology Life Quality Index score was 19.0 (Q1-Q3: 13.0-23.5). This score showed significant differences between patients with and without systemic symptoms; it demonstrated correlations with both depression and anxiety scores. Treatment costs caused financial hardship in 55.9% of patients, underscoring the burden associated with GPP. CONCLUSIONS The substantial disease and economic burdens among Chinese GPP patients warrant increased attention. Patients with early onset disease and those transitioning to plaque psoriasis require targeted interventions to mitigate the high recurrence risk.
Humans ; Male ; Female ; Psoriasis/pathology* ; Adult ; Cross-Sectional Studies ; Adolescent ; Child ; Young Adult ; Quality of Life ; Middle Aged ; China/epidemiology* ; Recurrence ; Risk Factors ; Surveys and Questionnaires ; East Asian People

BACKGROUND:D1-3-n-butylphthalide has antioxidant and anti-inflammatory effects and has been explored to have protective role in Parkinson's disease,but the underlying mechanisms are unknown.OBJECTIVE:To investigate the protective effect of D1-3-n-butylphthalide by the approach of network pharmacology,molecular docking,and cellular experimental validation.METHODS:(1)Network pharmacology and molecular docking:The database was used to screen the targets of D1-3-n-butylphthalide and Parkinson's disease.The intersection was taken from the construction of the target protein interaction network,and then screen the core targets.The GO and KEGG pathway enrichment was used to further analyze the core targets.The interaction between the target proteins and D1-3-n-butylphthalide was verified by molecular docking.(2)Cell validation:The passage 6 PC12 cells were divided into six groups for culture.The control group was cultured with conventional culture medium.The model group was cultured with N-methyl-4-phenylpyridinium iodide to induce Parkinson's disease model.The ML385 inhibitor group was added with nuclear factor E2-related factor 2 inhibitor ML385 on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide treatment group was added with butylphthalide on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide combined with ML385 treatment group was added with D1-3-n-butylphthalide and ML385 on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide group was cultured with conventional culture medium containing butylphthalide alone.Cell proliferation,intracellular reduced glutathione and malondialdehyde levels,and protein expression of protein kinase B/glycogen synthase kinase 3β/nuclear factor E2-related factor 2(AKT/GSK-3β/Nrf2)signaling pathway were detected.RESULTS AND CONCLUSION:(1)A total of 52 targets were screened for the intersection of drugs and disease targets,and the core targets including the matrix metalloproteinase 9 and GSK-3β were involved the phosphatidylinositol 3-kinase(PI3K)/AKT and oxidative stress-related signaling pathways.The molecular docking binding energy of D1-3-n-butylphthalide and GSK-3β was-18.27 kJ/mol,which indicated that D1-3-n-butylphthalide had a good binding ability with GSK-3β.(2)Compared with the model group,the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide treatment group were increased(P＜0.05),the malondialdehyde level was decreased(P＜0.05),and the expression of p-AKT,p-GSK-3β,Nu-Nrf2,and T-Nrf2 proteins was increased(P＜0.05).Compared with the D1-3-n-butylphthalide group,the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide combined with ML385 treatment group were decreased(P＜0.05),the malondialdehyde level was increased(P＜0.05),and the expression of Nu-Nrf2 and T-Nrf2 proteins was decreased(P＜0.05).(3)These results demonstrate that D1-3-n-butylphthalide can inhibit oxidative stress and improve cell activity through the AKT/GSK-3β/Nrf2 signaling pathway,and has a protective effect on the Parkinson's cell model induced by N-methyl-4-phenylpyridinium iodide.

BACKGROUND:The"inflammatory storm"induced by microglia is an important pathological factor causing the death of neuronal cells after spinal cord injury,which is extremely unfavorable for the recovery after spinal cord injury.Dihydroquercetin belongs to the vitamin P group and has excellent biological activity.It has good anti-inflammatory,anti-lipid peroxidation and neuroprotective effects on spinal cord injury.The mechanism by which it promotes the repair of spinal cord injury needs to be further clarified.OBJECTIVE:To investigate whether dihydroquercetin can ameliorate the inflammatory state induced after spinal cord injury through its anti-inflammatory and antioxidant properties for the purpose of protection against spinal cord injury.METHODS:Forty-eight Sprague-Dawley rats were divided into sham operation group,spinal cord injury group,low-dose dihydroquercetin group,and high-dose dihydroquercetin group according to the random number table method.The sham operation group only underwent laminectomy and was given intraperitoneal injection of normal saline after the operation.Animal models of spinal cord injury were made in the latter three groups.After the operation,normal saline,30 mg/kg,and 50 mg/kg dihydroquercetin were given by intraperitoneal injection respectively,and samples were collected 4 weeks after the operation.The Basso-Beatte-Bresnahan(BBB)limb motor function score and mechanical paw withdrawal threshold were used to evaluate the limb motor function and pain recovery of rats.Hematoxylin-eosin staining was used to observe the pathological changes of spinal cord tissue;Nissl staining was used to observe the number of Nissl bodies and the morphology of neurons.The activity of superoxide dismutase in spinal cord tissue was detected.Immunofluorescence was used to detect the expression of reactive oxygen species and microglia Iba1.Western blot was used to detect the protein expression of Iba1 and interleukin-1β,interleukin-6,interleukin-10,tumor necrosis factor-α,and p38 mitogen-activated protein kinase in the spinal cord.RESULTS AND CONCLUSION:(1)In the behavioral test,the BBB limb motor function score showed that since 14 days after the operation,the BBB limb motor function score of the high-dose dihydroquercetin group was significantly better than that of the spinal cord injury group(P<0.01);the mechanical paw withdrawal threshold was significantly lower in the high-dose dihydroquercetin than the spinal cord injury group at 21 days after the operation(P<0.05),and the former group showed better motor function.(2)Hematoxylin-eosin staining and Nissl staining showed that compared with the spinal cord injury group,the high-dose dihydroquercetin group showed slightly swollen but structurally intact and clearer neuron morphology in the spinal cord tissue,and the number of Nissl bodies increased.(3)Superoxide dismutase detection:Compared with the spinal cord injury group,the activity of superoxide dismutase in the spinal cord tissue of the high-dose dihydroquercetin increased significantly(P<0.05).(4)Reactive oxygen species immunofluorescence detection:Compared with the spinal cord injury group,the high-dose dihydroquercetin could significantly reduce the accumulation of reactive oxygen species after spinal cord injury(P<0.05).(5)Detection of microglial marker Iba1:Compared with the spinal cord injury group,the high-dose dihydroquercetin could significantly reduce the expression of microglial marker Iba1 after spinal cord injury(P<0.01).(6)Western blot detection results showed that compared with the spinal cord injury group,the expressions of inflammatory factors(interleukin-1β,interleukin-6,and tumor necrosis factor-α)and phosphorylated p38 mitogen-activated protein kinase in the high-dose dihydroquercetin group decreased significantly(P<0.01),while the expression of anti-inflammatory factor interleukin-10 increased significantly(P<0.01).These findings indicate that dihydroquercetin can effectively reverse the expression changes of multiple inflammatory markers induced after spinal cord injury in rats,thereby exerting neuroprotective effects and improving motor function.

BACKGROUND:D1-3-n-butylphthalide has antioxidant and anti-inflammatory effects and has been explored to have protective role in Parkinson's disease,but the underlying mechanisms are unknown.OBJECTIVE:To investigate the protective effect of D1-3-n-butylphthalide by the approach of network pharmacology,molecular docking,and cellular experimental validation.METHODS:(1)Network pharmacology and molecular docking:The database was used to screen the targets of D1-3-n-butylphthalide and Parkinson's disease.The intersection was taken from the construction of the target protein interaction network,and then screen the core targets.The GO and KEGG pathway enrichment was used to further analyze the core targets.The interaction between the target proteins and D1-3-n-butylphthalide was verified by molecular docking.(2)Cell validation:The passage 6 PC12 cells were divided into six groups for culture.The control group was cultured with conventional culture medium.The model group was cultured with N-methyl-4-phenylpyridinium iodide to induce Parkinson's disease model.The ML385 inhibitor group was added with nuclear factor E2-related factor 2 inhibitor ML385 on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide treatment group was added with butylphthalide on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide combined with ML385 treatment group was added with D1-3-n-butylphthalide and ML385 on the basis of inducing Parkinson's disease model.The D1-3-n-butylphthalide group was cultured with conventional culture medium containing butylphthalide alone.Cell proliferation,intracellular reduced glutathione and malondialdehyde levels,and protein expression of protein kinase B/glycogen synthase kinase 3β/nuclear factor E2-related factor 2(AKT/GSK-3β/Nrf2)signaling pathway were detected.RESULTS AND CONCLUSION:(1)A total of 52 targets were screened for the intersection of drugs and disease targets,and the core targets including the matrix metalloproteinase 9 and GSK-3β were involved the phosphatidylinositol 3-kinase(PI3K)/AKT and oxidative stress-related signaling pathways.The molecular docking binding energy of D1-3-n-butylphthalide and GSK-3β was-18.27 kJ/mol,which indicated that D1-3-n-butylphthalide had a good binding ability with GSK-3β.(2)Compared with the model group,the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide treatment group were increased(P＜0.05),the malondialdehyde level was decreased(P＜0.05),and the expression of p-AKT,p-GSK-3β,Nu-Nrf2,and T-Nrf2 proteins was increased(P＜0.05).Compared with the D1-3-n-butylphthalide group,the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide combined with ML385 treatment group were decreased(P＜0.05),the malondialdehyde level was increased(P＜0.05),and the expression of Nu-Nrf2 and T-Nrf2 proteins was decreased(P＜0.05).(3)These results demonstrate that D1-3-n-butylphthalide can inhibit oxidative stress and improve cell activity through the AKT/GSK-3β/Nrf2 signaling pathway,and has a protective effect on the Parkinson's cell model induced by N-methyl-4-phenylpyridinium iodide.

Objective:To explore molecular mechanism of high-altitude hypoxia regulates PPAR signaling pathway induced ferroptosis in spleen.Methods:Hypoxia animal model was constructed,target genes were screened and predicted by combination of transcriptomics and protein omics.Key genes in PPAR and ferroptosis pathways under hypoxia exposure were explored by GO and KEGG enrichment analysis and verified by RT-qPCR and Western blot.Results:Combination of transcriptomics and protein omics showed that 95 predicted target genes(protein)showed significantly differential expression under hypoxic exposure.GO annotation analysis and KEGG enrichment analysis showed that differential genes were mainly significantly enriched in PPAR and ferroptosis signaling pathways.A negative correlation was found between PPAR and ferroptosis signaling pathways,and GSEA showed that differential gene sets of PPAR and ferroptosis signaling pathways exhibited opposite expression trend in high-altitude hypoxia group.Validation of key genes PPARA,RXRB,APOA1 and SCD-1 in PPAR signaling pathway revealed that both mRNA and protein expres-sions were down-regulated under hypoxic exposure.Subsequently,differential expression was observed in mRNA and protein expres-sions of GPX4 in endogenous pathway and SLC7A11,TRP53 and TFRC in exogenous pathway in ferroptosis signaling pathway.Corre-lations between four key genes for ferroptosis and differential inflammation-associated genes(DE-IRGs)were positively or negatively.IL-1β,IL-6,IL-12,IL-18,IFN-γ and TNF-α expressions in spleen tissue were up-regulated under hypoxic exposure.Conclusion:High-altitude hypoxia exposure further induces ferroptosis through PPAR signaling pathway-mediated lipid metabolism disorders,and accompanied by occurrence of inflammatory response,which causes damage of spleen tissue.

BACKGROUND:The"inflammatory storm"induced by microglia is an important pathological factor causing the death of neuronal cells after spinal cord injury,which is extremely unfavorable for the recovery after spinal cord injury.Dihydroquercetin belongs to the vitamin P group and has excellent biological activity.It has good anti-inflammatory,anti-lipid peroxidation and neuroprotective effects on spinal cord injury.The mechanism by which it promotes the repair of spinal cord injury needs to be further clarified.OBJECTIVE:To investigate whether dihydroquercetin can ameliorate the inflammatory state induced after spinal cord injury through its anti-inflammatory and antioxidant properties for the purpose of protection against spinal cord injury.METHODS:Forty-eight Sprague-Dawley rats were divided into sham operation group,spinal cord injury group,low-dose dihydroquercetin group,and high-dose dihydroquercetin group according to the random number table method.The sham operation group only underwent laminectomy and was given intraperitoneal injection of normal saline after the operation.Animal models of spinal cord injury were made in the latter three groups.After the operation,normal saline,30 mg/kg,and 50 mg/kg dihydroquercetin were given by intraperitoneal injection respectively,and samples were collected 4 weeks after the operation.The Basso-Beatte-Bresnahan(BBB)limb motor function score and mechanical paw withdrawal threshold were used to evaluate the limb motor function and pain recovery of rats.Hematoxylin-eosin staining was used to observe the pathological changes of spinal cord tissue;Nissl staining was used to observe the number of Nissl bodies and the morphology of neurons.The activity of superoxide dismutase in spinal cord tissue was detected.Immunofluorescence was used to detect the expression of reactive oxygen species and microglia Iba1.Western blot was used to detect the protein expression of Iba1 and interleukin-1β,interleukin-6,interleukin-10,tumor necrosis factor-α,and p38 mitogen-activated protein kinase in the spinal cord.RESULTS AND CONCLUSION:(1)In the behavioral test,the BBB limb motor function score showed that since 14 days after the operation,the BBB limb motor function score of the high-dose dihydroquercetin group was significantly better than that of the spinal cord injury group(P<0.01);the mechanical paw withdrawal threshold was significantly lower in the high-dose dihydroquercetin than the spinal cord injury group at 21 days after the operation(P<0.05),and the former group showed better motor function.(2)Hematoxylin-eosin staining and Nissl staining showed that compared with the spinal cord injury group,the high-dose dihydroquercetin group showed slightly swollen but structurally intact and clearer neuron morphology in the spinal cord tissue,and the number of Nissl bodies increased.(3)Superoxide dismutase detection:Compared with the spinal cord injury group,the activity of superoxide dismutase in the spinal cord tissue of the high-dose dihydroquercetin increased significantly(P<0.05).(4)Reactive oxygen species immunofluorescence detection:Compared with the spinal cord injury group,the high-dose dihydroquercetin could significantly reduce the accumulation of reactive oxygen species after spinal cord injury(P<0.05).(5)Detection of microglial marker Iba1:Compared with the spinal cord injury group,the high-dose dihydroquercetin could significantly reduce the expression of microglial marker Iba1 after spinal cord injury(P<0.01).(6)Western blot detection results showed that compared with the spinal cord injury group,the expressions of inflammatory factors(interleukin-1β,interleukin-6,and tumor necrosis factor-α)and phosphorylated p38 mitogen-activated protein kinase in the high-dose dihydroquercetin group decreased significantly(P<0.01),while the expression of anti-inflammatory factor interleukin-10 increased significantly(P<0.01).These findings indicate that dihydroquercetin can effectively reverse the expression changes of multiple inflammatory markers induced after spinal cord injury in rats,thereby exerting neuroprotective effects and improving motor function.

Objective To explore the drug resistance mechanism and characteristics of carbopenem-resistant pseudo-monas aeruginosa(CRPA)in our hospital.Methods BD phoenix 100 automatic bacterial identification and drug sensitivity an-alyzer was used to identify and detect the drug sensitivity of the strains.The minimum inhibitory concentration(MIC)of ceftazi-dime/acibactam was detected by micro broth dilution method.The modified carbapenem inactivation method(mCIM)and colloi-dal gold immunochromatography were used to detect the carbapenemase phenotype of the strains.The whole genome sequencing was used to detect the carbapenemase resistance gene and ST typing of the screened positive strains.Results A total of 22 strains of clinically isolated CRPA were collected,of which the antibacterial drugs with the lowest resistance rate were ceftidine/avibatan(22.7% ),followed by gentamicin and amikacin(27.3% ),pyracillin/tazobactam(59.09% ),cefuroxime(63.6% ).Ceftazide and aminotransferrane(77.27% ),ciprofloxacin(86.36% ),levofloxacin(95.45% ).There are a total of 5 strains(22.7% )of carbapenems in 22 CRPA by phenotypic detection.The whole genome sequencing results show that 4 strains of ST549 CRPA carry metal β-lactamase IMP-45 and serine β-lactamase OXA-1,OXA-50,one strain is ST245 CRPA carries metal β-lactamase NDM-1,that is,all five CRPA strains produce metal β-lactamase.Conclusion The resistance rate of CRPA to ceftazidime/avibactam is low in our hospital.Carbapenemase-producing is not the main mechanism of CRPA resistance to car-bapenems,while metal β-Lactamase-producing is the main mechanism of CRPA resistance to ceftazidime avibactam.

Objective:To conduct a scoping review of research on the application of mobile health technology in self-management for stroke patients, identifying the main types, content elements, outcome indicators, and effectiveness of mobile health technology to provide references for related research and clinical practice.Methods:Based on the Joanna Briggs Institute's scoping review guidelines, a search was conducted in Cochrane Library, PubMed, CINAHL, Embase, Web of Science, China National Knowledge Infrastructure, Wanfang Data, and China Biology Medicine disc for Chinese and English literature on the use of mobile health technology in self-management of stroke patients, with a search timeframe up to December 31, 2023. Literature screening and data extraction were performed, and the included studies were summarized and analyzed.Results:A total of 18 studies were included. The main forms of mobile health technology used in stroke self-management included applications, social media, wearable devices, web platforms, and health management systems. Content areas covered knowledge provision and self-management skills, exercise management, speech management, medication management, daily activity management, health data monitoring, and communication with healthcare providers. Outcome indicators included self-management capability, physiological indicators, psychological indicators, feasibility indicators, and others.Conclusions:The application of mobile health technology in stroke self-management has shown promising results, but further verification is needed. Future work should establish an integrated "online+offline" intervention model, promote multidisciplinary team collaboration, focus on elderly patients, and conduct high-quality research to explore scientific, individualized mobile health technology intervention strategies to enhance disease recovery and improve quality of life.