Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

Aim To explore the effects of nuciferine on cognitive behavior and the underlying mechanisms,white matter injury(WMI),neuroinflammation,and ferroptosis in mice with chronic ischemic WMI.Meth-ods Sixty C57BL/6 mice were divided into a control group,a bilateral common carotid artery stenosis(BCAS)model group,and low/high-dose nuciferine groups(20/40 mg·kg-1).A chronic ischemic WMI model was established using BCAS surgery.Following eight weeks of treatment,cognitive behavior(Y-maze,novel object recognition,Morris water maze),white matter integrity(LFB/MBP staining),microglial acti-vation(Iba-1 immunofluorescence),inflammatory cy-tokines(ELISA for TNF-α,IL-1β,IL-6),ferroptosis markers(Fe2+,ROS,MDA,GSH),mitochondrial ultrastructure(electron microscopy),and protein ex-pression of the PI3K/Akt and NRF2/xCT/GPX4 signa-ling pathways(Western blot)were evaluated.Results Compared with the control group,the BCAS group showed significant cognitive decline(P＜0.05),re-duced myelin density,elevated inflammatory cytokines and ferroptosis markers(Fe2+,ROS,MDA),shrunk-en mitochondria,and downregulated PI3K/Akt and NRF2/xCT/GPX4 pathway proteins(P＜0.05).Nu-ciferine intervention significantly ameliorated these in-juries in BCAS mice,with the high-dose group exhibi-ting superior effects(P＜0.05).Conclusions Nu-ciferine exerts protective effects against chronic ische-mic WMI and cognitive impairment by activating the PI3K/Akt and NRF2/xCT/GPX4 signaling pathways,thereby suppressing neuroinflammation and ferroptosis.

Objective To evaluate whether the hand-drawn bronchial map can increase the endobronchial ultrasonography(EBUS)detection rate and the diagnosis rate for peripheral lung lesions.Methods We retrospectively analyzed 105 patients and divided them into hand-drawn navigation-EBUS group(n=71)and traditional navigation-EBUS group(n=34)according to different navigation methods.Collect case data on age,gender,lesion imaging characteristics(density,location,presence or absence of bronchial inflation sign),biopsy frequency,and final diagnosis from two groups of patients,and analyze the EBUS arrival rate,diagnosis rate,and influencing factors.Results Compared to that in the traditional navigation-EBUS group,the EBUS detection rate in the hand-drawn navigation-EBUS group was significantly higher.When lesions were>20 mm and≤30 mm,the diagnosis rate in the hand-drawn navigation-EBUS group was significantly higher than that in the traditional navigation-EBUS group.Multivariate Logistic regression analysis showed that lesions>20 mm(P=0.042)and bronchial generation of lesions≤5th generation(P=0.005)were favorable factors affecting the diagnosis rate of hand-drawn navigation-EBUS.Conclusion A hand-drawn bronchial map greatly increases the EBUS detection rate and diagnostic yield of bronchoscopy for peripheral lung lesions.

Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

Objective To evaluate whether the hand-drawn bronchial map can increase the endobronchial ultrasonography(EBUS)detection rate and the diagnosis rate for peripheral lung lesions.Methods We retrospectively analyzed 105 patients and divided them into hand-drawn navigation-EBUS group(n=71)and traditional navigation-EBUS group(n=34)according to different navigation methods.Collect case data on age,gender,lesion imaging characteristics(density,location,presence or absence of bronchial inflation sign),biopsy frequency,and final diagnosis from two groups of patients,and analyze the EBUS arrival rate,diagnosis rate,and influencing factors.Results Compared to that in the traditional navigation-EBUS group,the EBUS detection rate in the hand-drawn navigation-EBUS group was significantly higher.When lesions were>20 mm and≤30 mm,the diagnosis rate in the hand-drawn navigation-EBUS group was significantly higher than that in the traditional navigation-EBUS group.Multivariate Logistic regression analysis showed that lesions>20 mm(P=0.042)and bronchial generation of lesions≤5th generation(P=0.005)were favorable factors affecting the diagnosis rate of hand-drawn navigation-EBUS.Conclusion A hand-drawn bronchial map greatly increases the EBUS detection rate and diagnostic yield of bronchoscopy for peripheral lung lesions.

Objective To investigate the effect of naringenin(Nar)on hippocampal neuronal injury in neonatal mice with bilirubin encephalopathy,focusing on the Nrf2/GPX4-mediated ferroptosis pathway.Methods Neonatal mice were randomly divided into Control group,Model group,Nar low(Nar-L),high dose group(Nar-H)and high dose naringenin combined with Nrf2 inhibitor ML385 group(Nar-H+ML385),with 15 mice in each group.With the exception of the Control group,mice in the other groups were injected with bilirubin solution(20 μg/g)through the cerebellar bulbar cisterna to establish neonatal mouse bilirubin encephalopathy model.After the establishment of the model,intraperitoneal injection of naringin(25 or 100 mg/kg)or ML385(30 mg/kg)was administered once daily for a consecutive period of 7 days.After intervention,the neurobehavioral changes of each group of mice were observed.The water maze experiment was conducted to assess the long-term learning and memory abilities of the mice in each group.Nissl staining was performed to observe hippocampal neuron damage in mice.Chemical methods were used to measure the levels of malondialdehyde(MDA),4-hydroxynonenal(4-HNE),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)in the hippocampal tissue of mice.Colorimetric analysis was employed to determine the content of Fe2+in hippocampal tissue.Western blotting was utilized to detect protein expression levels of nuclear associated factor 2(Nrf2),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)protein in the hippocampal tissue of mice.Results Compared with the Control group,the Model group showed different degrees of abnormal neural behavior,and the long-term learning and memory ability were significantly reduced(P<0.05),at the same time,the hippocampal nerve was seriously damaged,and the contents of MDA,4-HNE and Fe2+in hippocampal tissue were significantly increased(P<0.05),the activities of SOD and GSH-Px and the protein expression levels of Nrf2,SLC7A11 and GPX4 were significantly decreased(P<0.05).Compared with the Model group,the Nar-L and Nar-H groups had less abnormal behavior,and the long-term learning and memory ability were significantly enhanced(P<0.05),at the same time,the hippocampal nerve injury was significantly improved,and the contents of MDA,4-HNE and Fe2+in hippocampal tissue were significantly decreased(P<0.05),the activities of SOD and GSH-Px and the protein expression levels of Nrf2,SLC7A11 and GPX4 were significantly increased(P<0.05).However,the combined intervention of ML385 significantly attenuates the beneficial effects of Nar on hippocampal neuron damage in neonatal mice with bilirubin encephalopathy.Conclusion This study suggested that Nar can ameliorate neuronal damage in hippocampus of neonatal mice with bilirubin encephalopathy,possibly through the regulation of iron death mediated by Nrf2/GPX4 axis.

Objective:To establish HPLC fingerprint of Shaoyao Gancao Keli to provide quality control.Methods:Waters Xselect CSH C18 column was used.The mobile phase was acetonitrile-0.05％phosphoric acid solution with gradient elution.The detection wavelength was 232 nm.The column temperature was 35 ℃.The flow rate was 0.8 mL·min-1.The injection volume was 10 μL.10 samples were tested by the method.The similarity of 10 samples were evaluated,and the common peaks were identified.Results:HPLC fingerprint of 17 common peaks was estab-lished.10 common peaks were identified which were gallic acid,albiflorin,paeoniflorin,liquiritin apioside,liquiritin,β-1,2,3,4,6-pentagalloylglucose,isoliquiritin apioside,isoliquiritin,liquiritigenin,glycyrrhizic acid.The similarity of 10 samples was higher than 0.95.Conclusion:The method has good separation,accuracy,re-peatability and stability,and could be used as a standard for quality control of Shaoyao Gancao Keli.

Objective:To establish HPLC fingerprint of Shaoyao Gancao Keli to provide quality control.Methods:Waters Xselect CSH C18 column was used.The mobile phase was acetonitrile-0.05％phosphoric acid solution with gradient elution.The detection wavelength was 232 nm.The column temperature was 35 ℃.The flow rate was 0.8 mL·min-1.The injection volume was 10 μL.10 samples were tested by the method.The similarity of 10 samples were evaluated,and the common peaks were identified.Results:HPLC fingerprint of 17 common peaks was estab-lished.10 common peaks were identified which were gallic acid,albiflorin,paeoniflorin,liquiritin apioside,liquiritin,β-1,2,3,4,6-pentagalloylglucose,isoliquiritin apioside,isoliquiritin,liquiritigenin,glycyrrhizic acid.The similarity of 10 samples was higher than 0.95.Conclusion:The method has good separation,accuracy,re-peatability and stability,and could be used as a standard for quality control of Shaoyao Gancao Keli.

Aim To explore the effects of nuciferine on cognitive behavior and the underlying mechanisms,white matter injury(WMI),neuroinflammation,and ferroptosis in mice with chronic ischemic WMI.Meth-ods Sixty C57BL/6 mice were divided into a control group,a bilateral common carotid artery stenosis(BCAS)model group,and low/high-dose nuciferine groups(20/40 mg·kg-1).A chronic ischemic WMI model was established using BCAS surgery.Following eight weeks of treatment,cognitive behavior(Y-maze,novel object recognition,Morris water maze),white matter integrity(LFB/MBP staining),microglial acti-vation(Iba-1 immunofluorescence),inflammatory cy-tokines(ELISA for TNF-α,IL-1β,IL-6),ferroptosis markers(Fe2+,ROS,MDA,GSH),mitochondrial ultrastructure(electron microscopy),and protein ex-pression of the PI3K/Akt and NRF2/xCT/GPX4 signa-ling pathways(Western blot)were evaluated.Results Compared with the control group,the BCAS group showed significant cognitive decline(P＜0.05),re-duced myelin density,elevated inflammatory cytokines and ferroptosis markers(Fe2+,ROS,MDA),shrunk-en mitochondria,and downregulated PI3K/Akt and NRF2/xCT/GPX4 pathway proteins(P＜0.05).Nu-ciferine intervention significantly ameliorated these in-juries in BCAS mice,with the high-dose group exhibi-ting superior effects(P＜0.05).Conclusions Nu-ciferine exerts protective effects against chronic ische-mic WMI and cognitive impairment by activating the PI3K/Akt and NRF2/xCT/GPX4 signaling pathways,thereby suppressing neuroinflammation and ferroptosis.

Objective:To explore the differences in bacterial communities within tissues during the process of oral mucosal carcinogenesis, and analyze the relationship between the high-abundance species Prevotella intermedia (Pi) and the occurrence and development of oral mucosal carcinogenesis. Methods:Fresh tissue samples were collected from patients diagnosed with oral leukoplakia (OLK), oral squamous cell carcinoma (OSCC), and healthy controls (HC) at Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, from January 2022 to November 2024, following strict inclusion criteria. Bacterial DNA was extracted from these specimens, and the 2bRAD sequencing for microbiome (2bRAD-M) was employed to analyze and compare the α and β diversity, as well as the community composition of bacteria within tissues, aiming to identify specifically expressed bacteria. Subsequently, paraffin-embedded clinical specimens were collected: 15 cases in the OLK group (including 4 cases of simple hyperplasia, 6 cases of mild dysplasia, and 5 cases of moderate to severe dysplasia), 12 cases in the OSCC group, and 5 cases in the HC group. A 4-nitroquinoline N-oxide (4NQO)-induced OLK progression mouse model was also constructed. Mice were randomly divided into three groups using a random number table, with six in each group. The negative control group was given distilled water to drink; Group 1 was given distilled water containing 4NQO to drink until week 12, while Group 2 was given distilled water containing 4NQO to drink until week 22. After the mice were sacrificed, their tongue tissue were collected and fixed. Fluorescence in situ hybridization (FISH) with specific probes was used to validate the presence of Pi in human and mouse tissue sections, analyzing the correlation between histopathological grading and the invasion depth of Pi.Results:The 2bRAD-M microbial analysis revealed that the relative abundance of Pi in OSCC tissues (10.80%) was significantly higher than in the HC group (0.50%) ( P=0.001) and OLK group (0.70%) ( P=0.002). FISH probe detection showed that the fluorescence intensity of Pi in human OSCC tissues [123.50 (101.00, 142.30)] was higher than in the HC group [0.00 (0.00, 28.50)], simple hyperplasia OLK [0.00 (0.00, 35.25)], and mild dysplasia OLK [24.50 (0.00, 55.50)] groups, with statistically significant differences respectively ( P=0.002, P=0.003, P=0.005). However, there was no significant difference compared to moderate to severe dysplasia OLK [56.00 (28.00, 62.50)] ( P=0.210). The fluorescence area of Pi in human OSCC tissues [8 615.00 (7 439.00, 11 084.00)] was significantly larger than in the HC group [0.00 (0. 00, 45.00)], simple hyperplasia OLK group [0.00 (0.00, 81.00)], mild dysplasia [49.00 (0.00, 151.00)], and moderate to severe dysplasia groups [1 450.00 (454.00, 2 892.00)], with highly significant differences ( P<0.001). There was a significant correlation between the invasive depth of Pi and the degree of histopathological grading ( P<0.001). In mice, the fluorescence intensity of Pi in OSCC tissues [120.00 (110.00, 127.00)] was significantly higher than in the HC group [0.00 (0.00, 12.25)] ( P<0.01), but showed no significant difference compared with the OLK group [50.00 (0.00, 58.00)] ( P>0.05). The fluorescence area of Pi in mice OSCC tissues [11 020.00 (6 790.00, 12 102.00)] was significantly larger than in the HC group [0.00 (0.00, 56.75)] and the OLK group [0.00 (0.00, 751.50)] ( P=0.006, P=0.043). There is a significant correlation between the depth of invasion of Pi and the degree of histopathological grading ( P<0.01). Conclusions:This study suggests that Pi in oral mucosal tissue may be a potential biomarker for early detection of OSCC and play an important role in the carcinogenesis process of oral mucosa.