Objective·To identify relevant biomarkers for patients with coronavirus disease 2019-associated kidney injury(COVID-19-associated KI)and explore the mechanisms underlying the involvement of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)proteins in infection-related KI by affecting the interactions between renal cells and macrophages.Methods·A retrospective analysis was conducted on the clinical characteristics of COVID-19 patients with KI treated in Shanghai Ninth,People's,hospital from December 2022 to February 2023.Serum levels of inflammatory factors and chemokines were measured by using enzyme-linked immunosorbent assay(ELISA).In vitro,human macrophage cell line THP-1 cells were stimulated with recombinant S1 subunit protein derived from SARS-CoV-2 spike protein.The cells and culture supernatants were collected to detect the levels of inflammatory factors and chemokines by using quantitative real-time PCR(qRT-PCR)and ELISA.Conditioned medium was prepared from the cell culture supernatants of S1-stimulated THP-1 cells and used to stimulate human renal epithelial cells(HK-2)in vitro to assess cytokine secretion.Antibody blocking experiments were performed to analyze the effects of the conditioned medium on the production of cytokines in HK-2 cells.Results·Among 39 patients with COVID-19,8(20.50%)had creatinine levels above the reference interval,which indicated the occurrence of KI.The levels of peripheral tumor necrosis factor-α(TNF-α)in the COVID-19 patient with KI group[(18.33±8.20)pg/mL]were significantly higher than those in the non-KI group[(11.88±6.50)pg/mL](P=0.015).In vitro assay has shown that S1-spike protein stimulation promoted the level of gene transcription and production of TNF-α,interleukin-1β(IL-1β)and chemokine C-X-C motif ligand 10(CXCL10)in THP-1 macrophage cells(P＜0.001).Furthermore,the conditioned medium from S1-stimulated THP-1 cells promoted the secretion of TNF-α,IL-1β and CXCL10 by HK-2 cells(P=0.005).When anti-TNF-α antibody(Infliximab)was used to block TNF-α in the culture supernatants from S1-stimulated THP-1 cells,the secretion level of TNF-α by HK-2 cells decreased dramatically(P＜0.001).Conclusion·TNF-α levels increase significantly in COVID-19 patients with KI,implying the significance of TNF-α in the occurrence of COVID-19-associated KI.In vitro experiments confirm that the S1 protein induces TNF-α secretion from THP-1 cells,leading to increased inflammatory responses in renal cells,which may contribute to the development of COVID-19-associated KI.Therefore,targeting TNF-α may become an alternative strategy to reduce the occurrence of COVID-19-associated KI.

Real-time acquisition of pulmonary ventilation and perfusion information through thoracic electrical impedance tomography (EIT) holds significant clinical value. This study proposes a novel method based on the rime (RIME) algorithm-optimized variational mode decomposition (VMD) to separate lung ventilation and perfusion signals directly from raw voltage data prior to EIT image reconstruction, enabling independent imaging of both parameters. To validate this approach, EIT data were collected from 16 healthy volunteers under normal breathing and inspiratory breath-holding conditions. The RIME algorithm was employed to optimize VMD parameters by minimizing envelope entropy as the fitness function. The optimized VMD was then applied to separate raw data across all measurement channels in EIT, with spectral analysis identifying relevant components to reconstruct ventilation and perfusion signals. Results demonstrated that the structural similarity index (SSIM) between perfusion images derived from normal breathing and breath-holding states averaged approximately 84% across all 16 subjects, significantly outperforming traditional frequency-domain filtering methods in perfusion imaging accuracy. This method offers a promising technical advancement for real-time monitoring of pulmonary ventilation and perfusion, holding significant value for advancing the clinical application of EIT in the diagnosis and treatment of respiratory diseases.
Humans ; Electric Impedance ; Algorithms ; Tomography/methods* ; Pulmonary Ventilation/physiology* ; Lung/diagnostic imaging* ; Image Processing, Computer-Assisted/methods* ; Adult

Objective:To establish a qualitative analysis of Rhodiola crenulata for determing six components inclu-ding gallic acid,salidroside,tyrosol,1,2,3,4,6-O-gallic glucose,rhodiosin,oxorlin-7-O-rhamnoside by HPLC,and to find out the feasibility of the method of linear calibration using two reference substances in qualitative analysis of chromatographic peaks.Methods:The real retention time of 6 components in Rhodiola crenulata on 19 chromatographic columns were determined.Gallic acid and rhodiosin were selected as the reference substances,and the method of linear calibration using these 2 substances was used to predict the retention time.Tyrosol was also chosen as the reference to predict the retention time with the relative retention time method(RRT method).Comparing the accuracy of these two methods.Results:Compared to the RRT method,the method of linear calibration with two reference substances was more accurate for predicting the retention time and more adapatable for many kinds of chromatographic columns.Conclusion:As a new alternative reference substance method,the method of linear calibration using two reference substances can assist chromatographic peak determination better and has broad application prospects.

Objective:To investigate the effects of pulsed electric field (PEF) combined with gemcitabine (GEM) on the viability and stemness of HCCC-9810 cholangiocarcinoma stem cells.Methods:HCCC-9810 cholangiocarcinoma stem cells were established in serum-free, cytokine-rich medium and divided into four groups: the control group, GEM group, PEF group, and the pulsed electric field combined with gemcitabine (PEF+ GEM) group. Cell proliferation was detected using the Cell Counting Kit-8 (CCK8) assay. Cell viability and apoptosis rate were measured by flow cytometry. Cell invasion ability was assessed using the Transwell assay. The expression of stemness marker proteins CD133 and Octamer-binding transcription factor 4 (OCT4), as well as the expression of β-catenin, was detected by Western blotting.Results:Regarding cell viability, the GEM, PEF, and PEF+ GEM groups showed significantly lower cell viability and higher apoptosis rate than the control group at 24 h, 48 h, and 72 h (all P<0.05). At 48 h and 72 h, the PEF+ GEM group showed significantly lower cell viability (7.2%±0.3% and 5.9%±0.8%, respectively) than the GEM group (50.7%±0.6% and 31.0%±1.2%, respectively) and the PEF group (12.2%±0.2% and 12.8%±0.2%, respectively) (all P<0.05). Regarding stemness inhibition, the PEF+ GEM groups showed significantly lower expression levels of CD133 and OCT4 at 24 h, 48 h, and 72 h compared with the control group (all P<0.05). Notably, at 48 h, the PEF+ GEM group showed a significantly lower expression level of the OCT4 (0.61±0.02) than the GEM group (0.87±0.08) and the PEF group (1.00±0.10) ( P<0.01). Furthermore, at 24 h and 48 h, the GEM, PEF, and PEF+ GEM groups showed significantly lower expression levels of β-catenin compared with the control group (all P<0.05). Conclusion:Pulsed electric field combined with gemcitabine therapy demonstrated more effective anti-proliferation and cancer stemness inhibition effects on HCCC-9810 cholangiocarcinoma stem cells compared with either monotherapy.

Background and aims:Primary sclerosing cholangitis(PSC)is an autoimmune liver disease characterized by complex pathogenesis and limited available therapeutic options.The mechanisms underlying the development and progression of PSCs remain unclear.Liver X receptor beta(LXR-β)is recognized to modulate lipid metabolism and immune response,but its specific involvement in the PSC has not been elucidated.Here,we explored the role and mechanism of LXR-β in PSC induced by 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine(DDC).Methods:CRISPR-Cas9 technology was applied to generate Abcb4(coding MDR2,next named as Mdr2),Nr1h2(coding LXR-β,next named as Lxrβ),and Rag2(coding RAG2)knockout mice.DDC was used to induce PSC.Hematoxylin and eosin and Sirius red staining were used to assess the extent of hepatic injury and fibrosis.Flow cytometry was used to observe immune cell subsets.Results:We observed a declining trend in hepatic Lxrβ in the PSC model.Unexpectedly,Lxrβ knockout failed to modulate DDC-induced PSC pathogenesis.Concomitantly,assessment of the influence of Rag2 deficiency on PSC progression revealed the absence of aggravated or alleviated hepatic injury or fibrosis in the Rag2-/-DDC mice.However,Lxrβ depletion intensified DDC-induced PSC in the Rag2-/-mice,with more abundant infiltrative inflammatory cells and more severe liver fibrosis.Compared with Rag2-/-DDC mice,Lxrβ-/-Rag2-/-DDC mice had higher serum ALT and AST levels and mRNA expression of proinflammatory and profibrotic genes.Flow cytometry showed that LXR-β deficiency resulted in a diminished population of hepatic innate immune cells.Conclusion:This study indicated innate immune cell LXR-β deficiency can exacerbate hepatic injury and fibrosis in murine models of PSC suggesting that LXR-β may regulate the function of innate immunity in the fibrotic advancement of PSC.

Objective·To identify relevant biomarkers for patients with coronavirus disease 2019-associated kidney injury(COVID-19-associated KI)and explore the mechanisms underlying the involvement of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)proteins in infection-related KI by affecting the interactions between renal cells and macrophages.Methods·A retrospective analysis was conducted on the clinical characteristics of COVID-19 patients with KI treated in Shanghai Ninth,People's,hospital from December 2022 to February 2023.Serum levels of inflammatory factors and chemokines were measured by using enzyme-linked immunosorbent assay(ELISA).In vitro,human macrophage cell line THP-1 cells were stimulated with recombinant S1 subunit protein derived from SARS-CoV-2 spike protein.The cells and culture supernatants were collected to detect the levels of inflammatory factors and chemokines by using quantitative real-time PCR(qRT-PCR)and ELISA.Conditioned medium was prepared from the cell culture supernatants of S1-stimulated THP-1 cells and used to stimulate human renal epithelial cells(HK-2)in vitro to assess cytokine secretion.Antibody blocking experiments were performed to analyze the effects of the conditioned medium on the production of cytokines in HK-2 cells.Results·Among 39 patients with COVID-19,8(20.50%)had creatinine levels above the reference interval,which indicated the occurrence of KI.The levels of peripheral tumor necrosis factor-α(TNF-α)in the COVID-19 patient with KI group[(18.33±8.20)pg/mL]were significantly higher than those in the non-KI group[(11.88±6.50)pg/mL](P=0.015).In vitro assay has shown that S1-spike protein stimulation promoted the level of gene transcription and production of TNF-α,interleukin-1β(IL-1β)and chemokine C-X-C motif ligand 10(CXCL10)in THP-1 macrophage cells(P＜0.001).Furthermore,the conditioned medium from S1-stimulated THP-1 cells promoted the secretion of TNF-α,IL-1β and CXCL10 by HK-2 cells(P=0.005).When anti-TNF-α antibody(Infliximab)was used to block TNF-α in the culture supernatants from S1-stimulated THP-1 cells,the secretion level of TNF-α by HK-2 cells decreased dramatically(P＜0.001).Conclusion·TNF-α levels increase significantly in COVID-19 patients with KI,implying the significance of TNF-α in the occurrence of COVID-19-associated KI.In vitro experiments confirm that the S1 protein induces TNF-α secretion from THP-1 cells,leading to increased inflammatory responses in renal cells,which may contribute to the development of COVID-19-associated KI.Therefore,targeting TNF-α may become an alternative strategy to reduce the occurrence of COVID-19-associated KI.

Objective·To explore the causal relationship between air pollution and the risk of Alzheimer's disease(AD)by using two-sample Mendelian randomization(MR).Methods·Based on the data from the genome-wide association study(GWAS),a two-sample MR analysis was conducted to evaluate the causal relationship between air pollution and the risk of AD.Air pollution indicators,including particulate matter 2.5(PM2.5),particulate matter 2.5-10(PM2.5-10),particulate matter 10(PM10),nitrogen dioxide and nitrogen oxides,were used as exposure factors,and summarized data were aggregated from the UK Biobank database.The PM2.5 dataset included 423 796 cases,with correlation analysis conducted on 9 851 867 single nucleotide polymorphisms(SNPs);the PM2.5-10 dataset included 423 796 cases,with correlation analysis conducted on 9 851 867 SNPs;the PM10 dataset included 455 314 cases,with correlation analysis conducted on 9 851 867 SNPs;the nitrogen dioxide dataset included 456 380 cases,with correlation analysis conducted on 9 851 867 SNPs;the nitrogen oxides dataset included 456 380 cases,with correlation analysis conducted on 9 851 867 SNPs.AD was used as the outcome factor,and data were obtained from the International Genomics of Alzheimer's Project(IGAP).The AD dataset included 25 580 cases and 48 466 controls,with correlation analysis of 7 067 513 SNPs.SNPs significantly associated with AD were used as instrumental variables.The main analysis was conducted by using the inverse variance weighted(IVW)method,and four methods including weighted median,MR-Egger regression,mode-based simple estimation and mode-based weighted estimation were used for quality control.Heterogeneity testing,gene pleiotropy testing and sensitivity analysis were conducted to assess the reliability of the study results.Results·Heterogeneity testing indicated no evidence of heterogeneity among SNPs associated with air pollution indicators and AD(both IVW and MR-Egger results,P＞0.05).Gene pleiotropy testing did not detect any pleiotropic effects(MR-Egger results,P＞0.05).Sensitivity analysis confirmed the stability of the PM2.5 results.IVW analysis revealed a statistically significant association between PM2.5 and AD in European populations(P＜0.001),while no statistically significant associations were observed between PM2.5-10(P=0.664),PM10(P=0.664),nitrogen dioxide(P=0.284),nitrogen oxides(P=0.567)and AD.Conclusion·There is a significant causal relationship between PM2.5 exposure and the risk of AD,with PM2.5 exposure increasing the incidence of AD.However,no evidence has been found to suggest that PM2.5-10,PM10,nitrogen dioxide or nitrogen oxides cause an increased risk of AD.

Objective:To establish a qualitative analysis of Rhodiola crenulata for determing six components inclu-ding gallic acid,salidroside,tyrosol,1,2,3,4,6-O-gallic glucose,rhodiosin,oxorlin-7-O-rhamnoside by HPLC,and to find out the feasibility of the method of linear calibration using two reference substances in qualitative analysis of chromatographic peaks.Methods:The real retention time of 6 components in Rhodiola crenulata on 19 chromatographic columns were determined.Gallic acid and rhodiosin were selected as the reference substances,and the method of linear calibration using these 2 substances was used to predict the retention time.Tyrosol was also chosen as the reference to predict the retention time with the relative retention time method(RRT method).Comparing the accuracy of these two methods.Results:Compared to the RRT method,the method of linear calibration with two reference substances was more accurate for predicting the retention time and more adapatable for many kinds of chromatographic columns.Conclusion:As a new alternative reference substance method,the method of linear calibration using two reference substances can assist chromatographic peak determination better and has broad application prospects.

Objective:To investigate the effects of pulsed electric field (PEF) combined with gemcitabine (GEM) on the viability and stemness of HCCC-9810 cholangiocarcinoma stem cells.Methods:HCCC-9810 cholangiocarcinoma stem cells were established in serum-free, cytokine-rich medium and divided into four groups: the control group, GEM group, PEF group, and the pulsed electric field combined with gemcitabine (PEF+ GEM) group. Cell proliferation was detected using the Cell Counting Kit-8 (CCK8) assay. Cell viability and apoptosis rate were measured by flow cytometry. Cell invasion ability was assessed using the Transwell assay. The expression of stemness marker proteins CD133 and Octamer-binding transcription factor 4 (OCT4), as well as the expression of β-catenin, was detected by Western blotting.Results:Regarding cell viability, the GEM, PEF, and PEF+ GEM groups showed significantly lower cell viability and higher apoptosis rate than the control group at 24 h, 48 h, and 72 h (all P<0.05). At 48 h and 72 h, the PEF+ GEM group showed significantly lower cell viability (7.2%±0.3% and 5.9%±0.8%, respectively) than the GEM group (50.7%±0.6% and 31.0%±1.2%, respectively) and the PEF group (12.2%±0.2% and 12.8%±0.2%, respectively) (all P<0.05). Regarding stemness inhibition, the PEF+ GEM groups showed significantly lower expression levels of CD133 and OCT4 at 24 h, 48 h, and 72 h compared with the control group (all P<0.05). Notably, at 48 h, the PEF+ GEM group showed a significantly lower expression level of the OCT4 (0.61±0.02) than the GEM group (0.87±0.08) and the PEF group (1.00±0.10) ( P<0.01). Furthermore, at 24 h and 48 h, the GEM, PEF, and PEF+ GEM groups showed significantly lower expression levels of β-catenin compared with the control group (all P<0.05). Conclusion:Pulsed electric field combined with gemcitabine therapy demonstrated more effective anti-proliferation and cancer stemness inhibition effects on HCCC-9810 cholangiocarcinoma stem cells compared with either monotherapy.

Objective·To explore the causal relationship between air pollution and the risk of Alzheimer's disease(AD)by using two-sample Mendelian randomization(MR).Methods·Based on the data from the genome-wide association study(GWAS),a two-sample MR analysis was conducted to evaluate the causal relationship between air pollution and the risk of AD.Air pollution indicators,including particulate matter 2.5(PM2.5),particulate matter 2.5-10(PM2.5-10),particulate matter 10(PM10),nitrogen dioxide and nitrogen oxides,were used as exposure factors,and summarized data were aggregated from the UK Biobank database.The PM2.5 dataset included 423 796 cases,with correlation analysis conducted on 9 851 867 single nucleotide polymorphisms(SNPs);the PM2.5-10 dataset included 423 796 cases,with correlation analysis conducted on 9 851 867 SNPs;the PM10 dataset included 455 314 cases,with correlation analysis conducted on 9 851 867 SNPs;the nitrogen dioxide dataset included 456 380 cases,with correlation analysis conducted on 9 851 867 SNPs;the nitrogen oxides dataset included 456 380 cases,with correlation analysis conducted on 9 851 867 SNPs.AD was used as the outcome factor,and data were obtained from the International Genomics of Alzheimer's Project(IGAP).The AD dataset included 25 580 cases and 48 466 controls,with correlation analysis of 7 067 513 SNPs.SNPs significantly associated with AD were used as instrumental variables.The main analysis was conducted by using the inverse variance weighted(IVW)method,and four methods including weighted median,MR-Egger regression,mode-based simple estimation and mode-based weighted estimation were used for quality control.Heterogeneity testing,gene pleiotropy testing and sensitivity analysis were conducted to assess the reliability of the study results.Results·Heterogeneity testing indicated no evidence of heterogeneity among SNPs associated with air pollution indicators and AD(both IVW and MR-Egger results,P＞0.05).Gene pleiotropy testing did not detect any pleiotropic effects(MR-Egger results,P＞0.05).Sensitivity analysis confirmed the stability of the PM2.5 results.IVW analysis revealed a statistically significant association between PM2.5 and AD in European populations(P＜0.001),while no statistically significant associations were observed between PM2.5-10(P=0.664),PM10(P=0.664),nitrogen dioxide(P=0.284),nitrogen oxides(P=0.567)and AD.Conclusion·There is a significant causal relationship between PM2.5 exposure and the risk of AD,with PM2.5 exposure increasing the incidence of AD.However,no evidence has been found to suggest that PM2.5-10,PM10,nitrogen dioxide or nitrogen oxides cause an increased risk of AD.