Fusobacterium nucleatum(F.nucleatum)is an early pathogenic colonizer in periodontitis,but the host response to infection with this pathogen remains unclear.In this study,we built an F.nucleatum infectious model with h...Fusobacterium nucleatum(F.nucleatum)is an early pathogenic colonizer in periodontitis,but the host response to infection with this pathogen remains unclear.In this study,we built an F.nucleatum infectious model with human periodontal ligament stem cells(PDLSCs)and showed that F.nucleatum could inhibit proliferation,and facilitate apoptosis,ferroptosis,and inflammatory cytokine production in a dose-dependent manner.The F.nucleatum adhesin Fad A acted as a proinflammatory virulence factor and increased the expression of interleukin(IL)-1β,IL-6 and IL-8.Further study showed that Fad A could bind with PEBP1 to activate the Raf1-MAPK and IKK-NF-κB signaling pathways.Time-course RNA-sequencing analyses showed the cascade of gene activation process in PDLSCs with increasing durations of F.nucleatum infection.NFκB1 and NFκB2 upregulated after 3 h of F.nucleatum-infection,and the inflammatory-related genes in the NF-κB signaling pathway were serially elevated with time.Using computational drug repositioning analysis,we predicted and validated that two potential drugs(piperlongumine and fisetin)could attenuate the negative effects of F.nucleatum-infection.Collectively,this study unveils the potential pathogenic mechanisms of F.nucleatum and the host inflammatory response at the early stage of F.nucleatum infection.展开更多
BACKGROUND：Paraquat （PQ） is a world-wide used herbicide and also a type of common poison for suicide and accidental poisoning. Numerous studies have proved that the concentration of serum PQ plays an important role...BACKGROUND：Paraquat （PQ） is a world-wide used herbicide and also a type of common poison for suicide and accidental poisoning. Numerous studies have proved that the concentration of serum PQ plays an important role in prognosis. Spectrophotometry, including common spectrophotometry and second-derivative spectrophotometry, is commonly used for PQ detection in primary hospitals. So far, lack of systematic research on the reliability of the method and the correlation between clinical features of patients with PQ poisoning and the test results has restricted the clinical use of spectrophotometry. This study aimed to evaluate the reliability and value of spectrophotometry in detecting the concentration of serum PQ. METHODS：The wavelengths for detecting the concentration of serum PQ by common and second-derivative spectrophotometry were determined. Second-derivative spectrophotometry was applied to detect the concentration of serum PQ. The linear range and precision for detection of PQ concentration by this method were confirmed. The concentration of serum PQ shown by second- derivative spectrophotometry and HPLC were compared in 8 patients with PQ poisoning. Altogether 21 patients with acute poisoning 4 hours after PQ ingestion treated in the period of October 2008 to September 2010 were retrospectively reviewed. The patients were divided into higher and lower than 1.8 μg/mL groups based on their concentrations of serum PQ measured by second-derivative spectrophotometry on admission. The severity of clinical manifestations between the two groups were analyzed with Student＇s t test or Fisher＇s exact test. RESULTS：The absorption peak of 257 nm could not be found when common spectrophotometry was used to detect the PQ concentration in serum. The calibration curve in the 0.4-8.0 μg/mL range for PQ concentration shown by second-derivative spectrophotometry obeyed Beer＇s law with r=0.996. The average recovery rates of PQ were within a range of 95.0% to 99.5%, relative standard deviation （RSD） was within 1.35% to 5.41% （n=6）, and the lower detection limit was 0.05 μg/mL. The PQ concentrations in serum of 8 patients with PQ poisoning shown by second-derivative spectrophotometry were consistent with the quantitative determinations by HPLC （r=0.995, P〈0.0001）. The survival rate was 22.2% in patients whose PQ concentration in serum was more than 1.8 μg/mL, and the incidences of acidosis, oliguria and pneumomediastinum in these patients were 55.6%, 55.6% and 77.8%, respectively. These clinical manifestations were different significantly from those of the patients whose PQ concentration in serum was less than 1.8 pg/mL （P〈0.05）. CONCLUSIONS： For common spectrophotometry, the wavelength at 257 nm was not suitable for detecting serum PQ as no absorbance was shown. Second-derivative spectrophotometry was reliable for detecting serum paraquat concentration. Serum PQ concentration detected by second- derivative spectrophotometry could be used to predict the severity of clinical manifestations of patients with PQ poisoning, and PQ content higher than 1.8 tJg/mL 4 hours after ingestion could be an important predictive factor for poor prognosis.展开更多
基金foundation support of the National Natural Science Foundation of China(Grant No.82071122)the Program of Taishan Young from Shandong Province+3 种基金Major Innovation Projects in Shandong Province(No.2021SFGC0502)Oral Microbiome Innovation Team of Young Scientist Project of Shandong Province(Grant No.2020KJK001)and Jinan City(2021GXRC021)The National High-level Young Scientist Project Foundation(2019)Excellent Young Scientist Foundation of Shandong Province(Grant No.ZR202102230369)。
文摘Fusobacterium nucleatum(F.nucleatum)is an early pathogenic colonizer in periodontitis,but the host response to infection with this pathogen remains unclear.In this study,we built an F.nucleatum infectious model with human periodontal ligament stem cells(PDLSCs)and showed that F.nucleatum could inhibit proliferation,and facilitate apoptosis,ferroptosis,and inflammatory cytokine production in a dose-dependent manner.The F.nucleatum adhesin Fad A acted as a proinflammatory virulence factor and increased the expression of interleukin(IL)-1β,IL-6 and IL-8.Further study showed that Fad A could bind with PEBP1 to activate the Raf1-MAPK and IKK-NF-κB signaling pathways.Time-course RNA-sequencing analyses showed the cascade of gene activation process in PDLSCs with increasing durations of F.nucleatum infection.NFκB1 and NFκB2 upregulated after 3 h of F.nucleatum-infection,and the inflammatory-related genes in the NF-κB signaling pathway were serially elevated with time.Using computational drug repositioning analysis,we predicted and validated that two potential drugs(piperlongumine and fisetin)could attenuate the negative effects of F.nucleatum-infection.Collectively,this study unveils the potential pathogenic mechanisms of F.nucleatum and the host inflammatory response at the early stage of F.nucleatum infection.
基金This study was supported by a grant from the National Natural Science Foundation of China（No.30871202）.
文摘BACKGROUND：Paraquat （PQ） is a world-wide used herbicide and also a type of common poison for suicide and accidental poisoning. Numerous studies have proved that the concentration of serum PQ plays an important role in prognosis. Spectrophotometry, including common spectrophotometry and second-derivative spectrophotometry, is commonly used for PQ detection in primary hospitals. So far, lack of systematic research on the reliability of the method and the correlation between clinical features of patients with PQ poisoning and the test results has restricted the clinical use of spectrophotometry. This study aimed to evaluate the reliability and value of spectrophotometry in detecting the concentration of serum PQ. METHODS：The wavelengths for detecting the concentration of serum PQ by common and second-derivative spectrophotometry were determined. Second-derivative spectrophotometry was applied to detect the concentration of serum PQ. The linear range and precision for detection of PQ concentration by this method were confirmed. The concentration of serum PQ shown by second- derivative spectrophotometry and HPLC were compared in 8 patients with PQ poisoning. Altogether 21 patients with acute poisoning 4 hours after PQ ingestion treated in the period of October 2008 to September 2010 were retrospectively reviewed. The patients were divided into higher and lower than 1.8 μg/mL groups based on their concentrations of serum PQ measured by second-derivative spectrophotometry on admission. The severity of clinical manifestations between the two groups were analyzed with Student＇s t test or Fisher＇s exact test. RESULTS：The absorption peak of 257 nm could not be found when common spectrophotometry was used to detect the PQ concentration in serum. The calibration curve in the 0.4-8.0 μg/mL range for PQ concentration shown by second-derivative spectrophotometry obeyed Beer＇s law with r=0.996. The average recovery rates of PQ were within a range of 95.0% to 99.5%, relative standard deviation （RSD） was within 1.35% to 5.41% （n=6）, and the lower detection limit was 0.05 μg/mL. The PQ concentrations in serum of 8 patients with PQ poisoning shown by second-derivative spectrophotometry were consistent with the quantitative determinations by HPLC （r=0.995, P〈0.0001）. The survival rate was 22.2% in patients whose PQ concentration in serum was more than 1.8 μg/mL, and the incidences of acidosis, oliguria and pneumomediastinum in these patients were 55.6%, 55.6% and 77.8%, respectively. These clinical manifestations were different significantly from those of the patients whose PQ concentration in serum was less than 1.8 pg/mL （P〈0.05）. CONCLUSIONS： For common spectrophotometry, the wavelength at 257 nm was not suitable for detecting serum PQ as no absorbance was shown. Second-derivative spectrophotometry was reliable for detecting serum paraquat concentration. Serum PQ concentration detected by second- derivative spectrophotometry could be used to predict the severity of clinical manifestations of patients with PQ poisoning, and PQ content higher than 1.8 tJg/mL 4 hours after ingestion could be an important predictive factor for poor prognosis.
