Objective To explore the mechanism by which zinc-regulated transporters, iron-regulated transporter-likeprotein 4 (ZIP4) regulates glycolysis and its impact on tumor progression in cholangiocarcinoma (CCA), providing a theoretical basis for targeted therapy of CCA. Methods ZIP4 expression in CCA was analyzed using the GEPIA database. Immuno-histochemistry (IHC) was used to detect ZIP4 expression in 20 paired CCA and adjacent non-tumor tissues. Stable ZIP4-overexpressing CCA cell lines (ZIP4-OE) were established. Gene set enrichment analysis was used to screen differentially expressed genes and pathways in ZIP-OE CCA cells. ZIP4, N-myc proto-oncogene protein (MYCN), and histone-lysine N-methyltransferase 2E (KMT2E) were knocked down using small interfering RNAs (siRNAs). The expression of glycolysis-related gene (glucose transporter 1 [Glut1], hexokinase 2 [HK2], and lactate dehydrogenase A [LDHA]) was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glycolytic activity was assessed by measuring the extracellular acidification rate (ECAR). Cell proliferation was evaluated using colony formation assays, and cell migration was assessed using Transwell assays. A xenograft mouse model was constructed to examine CCA tumor growth. Protein levels of ZIP4, KMT2E, H3K4me3 (tri-methylation of lysine 4 on histone H3), and MYCN were detected by Western blotting. Results GEPIA database analysis and IHC results confirmed significantly higher ZIP4 expression levels in CCA tissues compared to adjacent non-tumor tissues (P＜0.05). Compared to the control group, the ZIP4-OE group exhibited a significantly increased ECAR, along with significantly enhanced proliferation and migration abilities (P＜0.01). Conversely, knockdown of ZIP4 suppressed CCA cells proliferation and migration. GEPIA analysis indicated that ZIP4 upregulates the transcription of oncogene MYCN, as well as glycolysis-related genes. Knockdown of MYCN abolished the ZIP4 overexpression-induced upregulation of Glut1, HK2, and LDHA gene transcription, reduced glycolysis, and significantly inhibited CCA cell proliferation and migration (P＜0.05). Mechanistic studies demonstrated that ZIP4 increases H3K4me3 level via KMT2E, leading to MYCN transcription. Knockdown of KMT2E in CCA cells suppressed the ZIP4 overexpression-induced enhancement in H3K4me3 modification, resulting in MYCN downregulation and significantly reduced CCA cells proliferation and migration (P＜0.05). Conclusions ZIP4 upregulates H3K4me3 modification through KMT2E, which recruits transcription factors to activate the transcription of MYCN. This subsequently enhances cellular glycolysis and promotes the proliferation and migration of CCA cells.

BACKGROUND: Preclinical studies have indicated that Angong Niuhuang Pills (ANP) reduce cerebral infarct and edema volumes. This study aimed to investigate whether ANP safely reduces cerebral infarct and edema volumes in patients with moderate to severe acute ischemic stroke. METHODS: This randomized, double-blind, placebo-controlled pilot trial included patients with acute ischemic stroke with National Institutes of Health Stroke Scale (NIHSS) scores ranging from 10 to 20 in 17 centers in China between April 2021 and July 2022. Patients were allocated within 36 h after onset via block randomization to receive ANP or placebo (3 g/day for 5 days). The primary outcomes were changes in cerebral infarct and edema volumes after 14 days of treatment. The primary safety outcome was severe adverse events (SAEs) for 90 days. RESULTS: There were 57 and 60 patients finally included in the ANP and placebo groups, respectively for modified intention-to-treat analysis. The median age was 66.0 years, and the median NIHSS score at baseline was 12.0. The changes in cerebral infarct volume at day 14 were 0.3 mL and 0.4 mL in the ANP and placebo groups, respectively (median difference: -7.1 mL; interquartile range [IQR]: -18.3 to 2.3 mL, P = 0.30). The changes in cerebral edema volume of the ANP and placebo groups on day 14 were 11.4 mL and 4.0 mL, respectively ( median difference: 3.0 mL, IQR: -1.3 to 9.9 mL, P = 0.15). The rates of SAE within 90 days were similar in the ANP (3/57, 5%) and placebo (7/60, 12%) groups ( P = 0.36). Changes in serum mercury and arsenic concentrations were comparable. In patients with large artery atherosclerosis, ANP reduced the cerebral infarct volume at 14 days (median difference: -12.3 mL; IQR: -27.7 to -0.3 mL, P = 0.03). CONCLUSIONS: ANP showed a similar safety profile to placebo and non-significant tendency to reduce cerebral infarct volume in patients with moderate-to-severe stroke. Further studies are warranted to assess the efficacy of ANP in reducing cerebral infarcts and improving clinical prognosis. TRAIL REGISTRATION Clinicaltrials.gov , No. NCT04475328.
Aged ; Female ; Humans ; Male ; Middle Aged ; Double-Blind Method ; Drugs, Chinese Herbal/adverse effects* ; Ischemic Stroke/drug therapy* ; Pilot Projects ; Stroke/drug therapy* ; Treatment Outcome

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

Objective To find a reasonable frequency range of pure tone source signal and to propose a prelim-inary normal value determination range for sonotubometry for eustachian tube function assessment.Methods A to-tal of 31 healthy adults were enrolled in this study.Step pure tone line spectrum signals with an interval of 100 Hz on the frequency band of 3-6 kHz were used as sound sources for sonotubometry.Results The results showed that 8 frequency points of 3.5 kHz,4.3 kHz,4.35 kHz,4.5 kHz,4.65 kHz,4.8 kHz,5.0 kHz and 5.15 kHz were suitable frequency points for pure tone signal.The line spectrum data at these 8 frequency points were summarized and statistically analyzed,and 95％confidence interval of LVc value was calculated,that was,95％LVc value was greater than 5.5 dB,and 6 dB was set as the LVc threshold after integer.Conclusion In this study,a pure tone line spectrum sound source frequency suitable for sonotubometry was found,and the amplitude change of sound pressure level over 6 dB within about 0.5-1 second before and after swallowing action of the sound source signal can be used as an effective indicator of eustachian tube opening.

Objective:To evaluate long-term outcomes of endoscopic papillectomy (EP) for duodenal papillary adenomas and to identify risk factors for incomplete resection.Methods:Clinical data of 180 patients diagnosed as having duodenal papillary adenoma via postoperative pathology after EP in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January 2010 to December 2022 were retrospectively analyzed. Patients were divided into two groups based on their postoperative margin status: the complete resection group (negative resection margins) and the incomplete resection group (positive/uncertain resection margins). Recurrence rates were compared between the two groups, and logistic regression analysis was performed to identify risk factors for incomplete resection.Results:Among the 180 patients included in the study, 137 underwent complete resection, and 43 had incomplete resections. Recurrence rate was significantly higher in the incomplete resection group than that in the complete resection group (30.2% VS 15.3%, χ2=4.75, P=0.029). logistic regression analysis indicated that high-grade intraepithelial neoplasia was an independent risk factor for incomplete resection ( OR=2.43, 95% CI:1.12-5.26, P=0.024). Conclusion:Patients with incomplete resection after EP have a higher recurrence rate in the long-term follow-up. High-grade intraepithelial neoplasia is an independent risk factor for incomplete resection. Close surveillance and aggressive management are warranted for patients with positive or uncertain resection margins to mitigate the recurrence risk.

The elimination of biofilms is a crucial step in controlling hospital-acquired infections.Once biofilms colonize luminal instruments,it is difficult to remove them using traditional disinfection methods.Conventional disinfection approaches now face a series of challenges,including microbial resistance,corrosiveness,cytotoxicity,residual disinfection byproducts,and environmental pollution.Therefore,developing novel disinfection technologies specifically targeting biofilm removal is vitally important.New disinfection technologies,such as slightly acidic electrolyzed water,plasma technology,surface modification techniques,nanomaterial-based disinfection,bacteriophage disinfection,and enzymatic disinfection,are constantly emerging.These technologies exhibit excellent performance against biofilms by leveraging the synergistic effects of multiple mechanisms,including the reactive oxygen species(ROS)burst,photocatalytic oxidation,physical disruption,and biological targeting.This review summarizes the characteristics,underlying mechanisms,and potential application scenarios of these novel disinfection technologies,with a particular focus on their effects against biofilms formed by common pathogenic bacteria on surfaces in hospital settings.It aims to provide a reference basis for the practical application and translation of these disinfection technologies and the development of new disinfection strategies.

Objective To find a reasonable frequency range of pure tone source signal and to propose a prelim-inary normal value determination range for sonotubometry for eustachian tube function assessment.Methods A to-tal of 31 healthy adults were enrolled in this study.Step pure tone line spectrum signals with an interval of 100 Hz on the frequency band of 3-6 kHz were used as sound sources for sonotubometry.Results The results showed that 8 frequency points of 3.5 kHz,4.3 kHz,4.35 kHz,4.5 kHz,4.65 kHz,4.8 kHz,5.0 kHz and 5.15 kHz were suitable frequency points for pure tone signal.The line spectrum data at these 8 frequency points were summarized and statistically analyzed,and 95％confidence interval of LVc value was calculated,that was,95％LVc value was greater than 5.5 dB,and 6 dB was set as the LVc threshold after integer.Conclusion In this study,a pure tone line spectrum sound source frequency suitable for sonotubometry was found,and the amplitude change of sound pressure level over 6 dB within about 0.5-1 second before and after swallowing action of the sound source signal can be used as an effective indicator of eustachian tube opening.

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

Objective:To evaluate long-term outcomes of endoscopic papillectomy (EP) for duodenal papillary adenomas and to identify risk factors for incomplete resection.Methods:Clinical data of 180 patients diagnosed as having duodenal papillary adenoma via postoperative pathology after EP in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January 2010 to December 2022 were retrospectively analyzed. Patients were divided into two groups based on their postoperative margin status: the complete resection group (negative resection margins) and the incomplete resection group (positive/uncertain resection margins). Recurrence rates were compared between the two groups, and logistic regression analysis was performed to identify risk factors for incomplete resection.Results:Among the 180 patients included in the study, 137 underwent complete resection, and 43 had incomplete resections. Recurrence rate was significantly higher in the incomplete resection group than that in the complete resection group (30.2% VS 15.3%, χ2=4.75, P=0.029). logistic regression analysis indicated that high-grade intraepithelial neoplasia was an independent risk factor for incomplete resection ( OR=2.43, 95% CI:1.12-5.26, P=0.024). Conclusion:Patients with incomplete resection after EP have a higher recurrence rate in the long-term follow-up. High-grade intraepithelial neoplasia is an independent risk factor for incomplete resection. Close surveillance and aggressive management are warranted for patients with positive or uncertain resection margins to mitigate the recurrence risk.

Objective:To investigate the correlation between impulse oscillometry system examination indicators and conventional pulmonary ventilation function.Methods:The pulmonary ventilation function data of 10 883 patients from January 1, 2020 to December 31, 2022 at Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology were included. The one-second rate [ratio of forced expiratory volume in the first second (FEV 1) to forced vital capacity (FVC)] measured as a percentage of the predicted value was ≥92% for the control group ( n=3 478) and <92% for the pulmonary obstruction group ( n=7 405). The obstruction group was subdivided into five groups according to the degree of pulmonary dysfunction: mild group ( n=3 938),moderate group ( n=1 142),oderate-severe group ( n=917),severe group ( n=737),and extremely severe group ( n=671). Conventional pulmonary ventilatory function FVC, FEV 1, one-second rate, and forced expired flow at 50% of FVC (MEF50%), forced expired flow at 75% FVC (MEF25%), maximal mid-expiratory flow (MMEF), peak expiratory flow (PEF), and pulsed oscillation pulmonary function test were detected in both groups of patients. Impedance at 5 Hz (Z5) means total respiratory resistance, resistance at 5 Hz (R5) means total airway resistance, reactance at 5 Hz (X5) indicates the elastic recoil of the peripheral airways, and resistance at 20 Hz (R20) represents resistance of the central airways. R5-R20 reflects resistance in the small airways. Additionally, peripheral resistance (Rp), respiratory resonance frequency (Frex), and area under the reactance curve (Ax) were also measured. Correlation between the indicators of the two groups and the sensitivity and specificity of the impulse oscillometry system parameters for the diagnosis of obstructive pulmonary ventilation dysfunction were analyzed. Results:Pulmonary function force expiratory volume in the first second as a percentage of predicted value (FEV 1%Pre) [80.10 (54.95,97.10)%],one-second rate [62.43(48.67, 67.02)%],MEF50% [1.33 (0.62,1.97)L/s],MEF25% [0.28 (0.17,0.41)L/s], MMEF [0.85 (0.43,1.29)L/s],and PEF [5.64 (3.73,7.50)]L/s in the obstruction group were significantly lower than those in the control group ( P<0.05). The differences within the subgroups of the obstruction group were also significant ( P<0.05). Pulsed oscillation Z5 [0.42 (0.33,0.55)kPa·L -1·s -1],Rp [0.25 (0.20,0.45)kPa·L -1·s -1], R5 [0.39 (0.31,0.49)kPa·L -1·s -1], R20 [0.28 (0.24,0.34)kPa·L -1·s -1], R5-R20 [0.09 (0.05,0.17)kPa·L -1·s -1],Frex [16.32 (13.07,20.84)Hz], and Ax [0.67 (0.28,1.64)] indices in the obstruction group were significantly higher than those in the control group. X5 [-0.14 (-0.23, -0.10)kPa·L -1·s -1] was significantly lower than that in the control group ( P<0.05). Z5, Rp, X5, R5, R5-R20, Frex, and Ax were statistically significant between different degrees of obstruction in the obstruction group ( P<0.05). The impulse oscillometry system parameters Z5, Rp, R5, R20, R5-20, Frex, and Ax were negatively correlated with the indices of conventional pulmonary ventilation ( r=-0.21-0.68, P<0.05), and the parameter X5 was positively correlated with the indices of conventional pulmonary ventilation ( r=0.41-0.68, P<0.05). The pulsed oscillation pulmonary function test parameters X5 (58.60%-95.68%) and Ax (57.08%-98.06%) presented the best sensitivity; X5 (86.29%-98.82%), Frex (86.69%-94.71%), and Ax (88.10%-98.53%) displayed the best specificity; and R20 presented the worst sensitivity and specificity. The sensitivity and specificity were slightly better in female patients than in male patients. Conclusion:The technical parameters of the impulse oscillometry system showed significant correlation with relevant indices of conventional pulmonary ventilation function detection. These well reflect the changes of different degrees of pulmonary ventilation function and have greater significance for reference in evaluating the degree of pulmonary function impairment.