ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.

Objective To explore the effect of the ratio of high-density lipoprotein cholesterol(HDL-C)/total cholesterol(TC)on no-reflow after percutaneous coronary intervention(PCI)in elderly patients with acute coronary syndrome(ACS)complicated with diabetes mellitus(DM).Methods A retrospective analysis was conducted on 206 elderly ACS patients complicated with DM undergoing PCI in our hospital from January 2018 to August 2024.The HDL-C and TC levels were detected by cholesterol oxidase test,and the HDL-C/TC ratio was calculated.Coronary angi-ography(CAG)was applied to evaluate no-reflow phenomenon after PCI,and according to the re-sults,the patients were divided into a non-reflow group(41 cases)and a normal reflow group(165 cases).ROC curve was plotted to evaluate the predictive performance of HDL-C/TC ratio for no-reflow after PCI in patients with ACS complicated DM.Results The no-reflow group had signifi-cantly higher TC and glycated hemoglobin A1c(HbA1c)levels and more balloon dilatations,but lower HDL-C level and HDL-C/TC ratio than the normal flow group(P＜0.01).Multivariate lo-gistic regression analysis showed that HbA1c(OR=3.196,95％CI:1.619-6.310,P=0.001),number of balloon dilatations(OR=3.504,95％CI:1.807-6.797,P=0.000),and HDL-C/TC ra-tio(OR=3.927,95％CI:2.0 73-7.441,P=0.000)were influencing factors of no-reflow after PCI in patients with ACS and DM.The AUC value of HDLC,TC,and HDL-C/TC ratio in predicting no-reflow after PCI was 0.842,0.726,and 0.922,respectively.Conclusion HDL-C/TC ratio is an influencing factor for no-reflow in patients with ACS and DM after PCI.The ratio at a cut-off val-ue of ≤0.21 has a certain predictive value for no-reflow after PCI in these elderly patients.

Objective To explore the effect of the ratio of high-density lipoprotein cholesterol(HDL-C)/total cholesterol(TC)on no-reflow after percutaneous coronary intervention(PCI)in elderly patients with acute coronary syndrome(ACS)complicated with diabetes mellitus(DM).Methods A retrospective analysis was conducted on 206 elderly ACS patients complicated with DM undergoing PCI in our hospital from January 2018 to August 2024.The HDL-C and TC levels were detected by cholesterol oxidase test,and the HDL-C/TC ratio was calculated.Coronary angi-ography(CAG)was applied to evaluate no-reflow phenomenon after PCI,and according to the re-sults,the patients were divided into a non-reflow group(41 cases)and a normal reflow group(165 cases).ROC curve was plotted to evaluate the predictive performance of HDL-C/TC ratio for no-reflow after PCI in patients with ACS complicated DM.Results The no-reflow group had signifi-cantly higher TC and glycated hemoglobin A1c(HbA1c)levels and more balloon dilatations,but lower HDL-C level and HDL-C/TC ratio than the normal flow group(P＜0.01).Multivariate lo-gistic regression analysis showed that HbA1c(OR=3.196,95％CI:1.619-6.310,P=0.001),number of balloon dilatations(OR=3.504,95％CI:1.807-6.797,P=0.000),and HDL-C/TC ra-tio(OR=3.927,95％CI:2.0 73-7.441,P=0.000)were influencing factors of no-reflow after PCI in patients with ACS and DM.The AUC value of HDLC,TC,and HDL-C/TC ratio in predicting no-reflow after PCI was 0.842,0.726,and 0.922,respectively.Conclusion HDL-C/TC ratio is an influencing factor for no-reflow in patients with ACS and DM after PCI.The ratio at a cut-off val-ue of ≤0.21 has a certain predictive value for no-reflow after PCI in these elderly patients.

Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

OBJECTIVE To investigate the effects and potential mechanism of paeoniflorin on oxidative stress of ulcerative colitis(UC)mice based on adenosine monophosphate-activated protein kinase(AMPK)/nuclear factor-erythroid 2-related factor 2(Nrf2)pathway.METHODS Male BALB/c mice were randomly divided into control group,model group,inhibitor group(AMPK inhibitor Compound C 20 mg/kg),paeoniflorin low-,medium-and high-dose groups(paeoniflorin 12.5,25,50 mg/kg),high-dose of paeoniflorin+inhibitor group(paeoniflorin 50 mg/kg+Compound C 20 mg/kg),with 8 mice in each group.Except for the control group,mice in all other groups were given 4%dextran sulfate sodium solution for 5 days to establish the UC model.Subsequently,mice in each drug group were given the corresponding drug solution intragastrically or intraperitoneally,once a day,for 7 consecutive days.The changes in body weight of mice were recorded during the experiment.Twenty-four hours after the last administration,colon length,malondialdehyde(MDA)content,and activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in colon tissues were measured;histopathological morphology of colon tissues,tight junctions between intestinal epithelial cells,and histopathological scoring were all observed and evaluated;the mRNA expressions of AMPK and Nrf2,as well as the protein expressions of heme oxygenase-1(HO-1),occludin and claudin-1,were all determined in colon tissue.RESULTS Compared with model group,paeoniflorin groups exhibited recovery from pathological changes such as inflammatory cell infiltration and crypt damage in the colon tissue,as well as improved tight junction damage between intestinal epithelial cells.Additionally,significant increases or upregulations were observed in body weight,colon length,activities of SOD and GSH-Px,phosphorylation level of AMPK,and protein expression of Nrf2,HO-1,occludin,claudin-1,and mRNA expressions of AMPK and Nrf2;concurrently,MDA content and histopathological scores were significantly reduced(P＜0.05 or P＜0.01).In contrast,the inhibitor group showed comparable(P＞0.05)or worse(P＜0.05 or P＜0.01)indicators compared to the model group.Conversely,the addition of AMPK inhibitor could significantly reverse the improvement of high-dose paconiflorin(P＜0.01).CONCLUSIONS Paeoniflorin can repair intestinal epithelial cell damage in mice,improve tight junctions between epithelial cells,upregulate the expression of related proteins,and promote the expression and secretion of antioxidant-promoting molecules,thereby ameliorating UC;its mechanism may be associated with activating AMPK/Nrf2 antioxidant pathway.

Objective:To establish a rapid and accurate method for the detection of Klebsiella pneumoniae carbapenemase (KPC) carbapenemase gene based on recombinase aided amplification (RAA)-CRISPR-Cas13a (CRISPR-Cas13a) technology. Methods:Twenty-five clinical isolates of carbapenem-resistant Klebsiella pneumoniae (CRKP) and five carbapenem-sensitive Klebsiella pneumoniae (CSKP) strains preserved in 2020-2021 in Beijing Chuiyangliu Hospital were randomly collected, and the total DNA samples of the strains was extracted. RAA primers specific for KPC DNA and CRISPR RNA (crRNA) were designed to establish a rapid and accurate method for the detection of KPC carbapenemase gene based on RAA-CRISPR-Cas13a technology. The method was evaluated by plasmids and clinical sample strains, and the detection was also performed by Quantitative real-time PCR (qPCR) method to compare the detection rate and consistency of the two methods. Results:The RAA-CRISPR-Cas13a method can detect KPC plasmids and samples with a sensitivity of 1 copy/μl, which is higher than that of qPCR (10 1 copies/μl). Among the 30 clinical strains (including 25 CRKP strains and 5 CSKP strains), 23 strains were detected to carry KPC gene by both RAA-CRISPR-Cas13a method and qPCR method, and 7 strains were not detected with KPC gene. The detection rate of KPC gene in the 25 CRKP strains was 92% (23/25). The positive coincidence rate of the two methods was 100% (23/23). Conclusions:This study combined RAA amplification technology with CRISPR-Cas13a technology to establish a rapid and accurate method for detecting KPC carbapenemase gene. The method is useful for accurate screening of KPC carbapenemase-producing strains. It has a wide application prospect in drug resistance monitoring and infection control.

Liver regeneration plays a crucial role in the recovery after liver injury induced by N-acetyl-p-aminophenol(APAP).After APAP overdose,the degree of regeneration increases with the extent of liver injury,leading to the resolution of liver injury and spontaneous recovery in most cases.However,severe APAP overdose can impair liver regeneration and result in uncontrolled liver injury,even failure to recover or death in severe cases.Following APAP-induced liver injury,interactions between cells in the liver are essential for regenerative response.Liver regeneration is jointly regulated by multiple proliferative signaling pathways,involving various kinases,nuclear receptors,transcription factors,and coactivators.Severe APAP overdose can inhibit the activation of proliferative signaling pathways,thereby causing cell cycle arrest and impairing liver regeneration.Although liver regeneration plays a critical role in the repair of APAP-induced liver injury,the underlying mechanisms remain unclear.This article reviews the research advances in the role of liver regeneration in APAP-induced liver injury,in order to provide a reference for further basic research in this area.