Objective To explore the application of plasma 7 microRNA (miR7) testing in the differential diagnosis of very early-stage hepatocellular carcinoma (HCC). Methods This study is a multicenter real-world study. Patients with single hepatic lesion (maximum diameter≤2 cm) who underwent plasma miR7 testing at Zhongshan Hospital, Fudan University, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Anhui Provincial Hospital, and Peking University People’s Hospital between January 2019 and December 2024 were retrospectively enrolled. Patients were divided into very early-stage HCC group and non-HCC group, and the clinical pathological characteristics of the two groups were compared. The value of plasma miR7 levels, alpha-fetoprotein (AFP), and des-gamma-carboxy prothrombin (DCP) in the differential diagnosis of very early-stage HCC was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). In patients with both negative AFP and DCP (AFP＜20 ng/mL, DCP＜40 mAU/mL), the diagnostic value of plasma miR7 for very early-stage HCC was analyzed. Results A total of 64 528 patients from 4 hospitals underwent miR7 testing, and 1 682 were finally included, of which 1 073 were diagnosed with very early-stage HCC and 609 were diagnosed with non-HCC. The positive rate of miR7 in HCC patients was significantly higher than that in non-HCC patients (67.9% vs 24.3%, P＜0.001). ROC curves showed that the AUCs for miR7, AFP, and DCP in distinguishing HCC patients from the non-HCC individuals were 0.718, 0.682, and 0.642, respectively. The sensitivities were 67.85%, 43.71%, and 44.45%, and the specificities were 75.70%, 92.78%, and 83.91%, respectively. The pairwise comparison of AUCs showed that the diagnostic efficacy of plasma miR7 detection was significantly better than that of AFP or DCP (P＜0.05). Although its specificity was slightly lower than AFP and DCP, the sensitivity was significantly higher. Among patients negative for both AFP and DCP, miR7 maintained an AUC of 0.728 for diagnosing very early-stage HCC, with 67.82% sensitivity and 77.73% specificity. Conclusions Plasma miR7 testing is a potential molecular marker with high sensitivity and specificity for the differential diagnosis of small hepatic nodules. In patients with very early-stage HCC lacking effective molecular markers (negative for both AFP and DCP), miR7 can serve as a novel and effective molecular marker to assist diagnosis.

Objective To observe the expressions of nucleotide-binding oligomerization domain receptor 1(NOD1)and its mediated RIP2/NF-κB signaling pathway-related proteins and autophagy-related proteins in cerebral cortex of rats with cerebral ischemia-reperfusion injury(CIRI);To explore the possible mechanism of eye acupuncture alleviating CIRI.Methods SPF-grade male Wistar rats were randomly divided into blank control group(12 rats),sham-operation group(12 rats)and modeling group(36 rats).A CIRI model was established by improved suture method.The rats in the modeling group were randomly divided into the model group,eyeacupuncture group,outside the acupoint area group,with 12 rats in each group.Neurological deficits in rats were evaluated by Longa score,TTC staining was used to observe the cerebral infarction,HE staining was used to observe the morphology of ischemic brain tissue,electron microscopy was used to observe the formation of autophagosome in ischemic brain tissue,RT-qPCR was used to detect the mRNA expressions of NOD1,receptor interacting protein 2(RIP2),nuclear factor-κB(NF-κB)p65 in ischemic cerebral cortex,Western blot was used to detect the expressions of NOD1,RIP2,NF-κB p65 and autophagy related proteins in ischemic cerebral cortex.Results Compared with the sham-operation group,the neurological deficit score of rats in the model group significantly increased(P<0.01),the infarct volume significantly increased(P<0.01),the typical cribriform infarct foci and multiple autophagosomes appeared in the ischemic brain tissue,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly increased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly increased(P<0.01),and the protein expression of p62 significantly decreased(P<0.01).Compared with the model group,the neurological deficit score in eye acupuncture group significantly decreased(P<0.01),the cerebral infarction volume significantly decreased(P<0.01),the area of cribriform reticular infarct and the number of autophagosomes in ischemic brain tissue significantly decreased,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly decreased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly decreased(P<0.01),and the protein expression of p62 significantly increased(P<0.01).There was no statistical significance compared with outside the acupoint area group.Conclusion Eye acupuncture can attenuat the injury of neurons in CIRI rats,and its mechanism may be related to inhibiting the activation of RIP2/NF-κB signaling pathway mediated by NOD1,thereby reducing autophagy of neurons.

Objective:To investigate the protective effects of Mailuoning(MLN)against cisplatin(CP)-induced acute kidney injury(AKI)utilizing network pharmacology and to analyze the underlying mechanism of action related to anti-apoptotic pathways.Methods:Active components of MLN and targets related to AKI were identified using network pharmacology.The active components of MLN were sourced from the TCMSP and ETCM 2.0 databases.The targets of components were predicted using the Swiss Target Prediction tool,and subsequently the targets related to AKI were retrieved from the GeneCards database to identify intersecting targets.A protein-protein interaction network was constructed using the STRING database,and a topological analysis was performed using Cytoscape to identify core targets.Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway en-richment analyses were conducted on these core targets.For the animal experiments,forty mice were randomly assigned to four groups:solvent control,MLN toxicity control,CP model,and CP+MLN groups.The MLN group received intraperitoneal injections of MLN at a dose of 15 mL/(kg·d)for ten consecutive days.The CP model and CP+MLN groups were administered a single intraperitoneal injec-tion of CP at 20 mg/kg on day 7.Three days after the CP treatment,plasma levels of blood urea nitrogen(BUN)and creatinine(Cre)were measured.The pathological injury of kidney tissues was as-sessed using hematoxylin-eosin staining.Western blot analysis was utilized to determine the expression of neutrophil gelatinase-associated lipocalin(NGAL)and apoptosis-related proteins in kid-ney tissues.Results:A total of 104 active components of MLN and 224 targets were identified using network pharmacology,and 2 465 targets were identified to be related to AKI,resulting in 117 intersecting targets,of which,17 targets were classified as core targets.KEGG and GO analyses indicated that the apoptosis-related signal transduction pathway might be a crucial pathway through which MLN provided protective effects against AKI.The results of animal experiments confirmed the successful establishment of CP-induced AKI models in mice.Compared with the CP model group,MLN treatment significantly reduced plasma levels of BUN and Cre(P<0.05),inhibited NGAL protein expression in the kidneys(P<0.05),and improved the pathological injury observed in kidney tissues.Furthermore,MLN markedly reduced the expression levels of p-P53(ser 15)and cleaved caspase-3 proteins,as well as the Bax/Bcl-2 ratio in kidney tissues of AKI model mice(P<0.05),while upregulating protein kinase B phosphorylation levels(P<0.05).Conclusion:MLN demonstrates protective effects against CP-induced AKI in mice,potentially through mechanisms related to its anti-apoptotic properties.

Abstract Oral cavity cancer is the sixth-leading cancer worldwide, which has become one of the important factors affecting oral health. The morbidity of oral squamous cell carcinoma(OSCC) accounts for nearly 80% of oral cavity cancer. The pathogenesis and prevention of OSCC have become a hot topic in this field. Previous studies have shown that bone morphogenetic proteins(BMPs) participated in the development, invasion and metastasis of gastric and bone cancers, thereby impacting the prognosis of the patients. Recent studies have found that BMPs also play a key role in the development, progression, metastasis, and invasion of OSCC, This review therefore summarizes the correlation and molecular mechanism between BMPs and the biological behavior of OSCC.

To investigate the molecular mechanisms underlying the mechanosensitive ion channel PI-EZO2 in osteoarthritis(OA),we developed a lentiviral vector for endogenous PIEZO2 overexpression and established a stable PIEZO2-high-expressing immortalized human primary chondrocyte line.By map-ping the open reading frame of the PIEZO2 locus and designing sequence-specific sgRNA,we employed the CRISPR/Cas9 synergistic activation mediator(SAM)system to precisely integrate transcriptional ac-tivation elements into the PIEZO2 promoter region.Lentiviral-mediated targeted genomic integration en-sured endogenous PIEZO2 overexpression,confirmed by mCherry fluorescence tracing coupled with flow cytometric sorting,which revealed membrane-specific localization of PIEZO2 protein(localization effi-ciency:78.49％).Quantitative PCR demonstrated a 17-fold upregulation of PIEZO2 mRNA,while Western blotting validated enhanced membrane-localized protein expression.Strikingly,PIEZO2-overex-pressing chondrocytes exhibited hallmark OA metabolic phenotypes compared to wild-type controls:typeⅡ collagen mRNA expression decreased to 50％of baseline levels,whereas matrix metalloproteinase 13(MMP13)mRNA surged by 20-fold.These alterations recapitulated the pathological matrix metabolic phenotype observed in biomechanical OA models induced by cyclic mechanical stress(10％strain,0.5 Hz,8 h/day for 2 consecutive days).Collectively,we successfully generated a human chondrocyte model with stable PIEZO2 overexpression,which faithfully mirrors mechanotransduction-driven OA progression.This engineered cellular system provides a robust platform for dissecting PIEZO2-mediated mechanosig-naling networks and advancing targeted therapeutic discovery.

Objective To investigate the coagulation effect of a cold atmospheric plasma(CAP)jet device with helium as the working gas and to study its coagulation mechanism preliminarily.Methods A CAP jet device treatment group,a helium airflow treatment group,a hot air treatment group(60℃)and a natural coagulation group were formed according to the treatment modes of the blood samples,with 10 μL of blood samples involved in each group,in order to validate the coagulation effect of the CAP jet device in vitro;the coagulation mechanism of the CAP jet device was explored by its application to the treatment of anticoagulated whole blood,platelet-rich plasma and platelet-depleted plasma;the coagulation effect of the CAP jet device in vivo was verified with a mouse liver punctate hemorrhage model and a rabbit mesenteric hemorrhage model.Results The CAP jet device can significantly accelerate the coagulation of anticoagulated blood droplets,and the coagulation time of anticoagulated blood droplets in the CAP jet device-treated group was shortened from 28 min in the natural coagulation group to(23±1.56)s,with the difference statistically significant(P<0.05),and the CAP jet device treatment group gained advantages significantly over the helium airflow treatment group(P<0.05)and the hot air(60℃)treatment group(P<0.05)in coagulation-promoting effect;the procoagulant effect of the CAP jet device rose with the increase of platelet content in blood droplets,and the coagulation effect of platelet-rich blood droplets was significantly better than that of whole blood(P<0.05),while no coagulation was observed in platelet-poor droplets.The CAP jet device could rapidly stop hemostasis of punctate hemorrhage in mouse liver and mesenteric hemorrhage in rabbits without delayed hemorrhage occurring within 10 min,and no obvious structural abnormality of the liver and thermal damage of the tissue were found microscopically.Conclusion The CAP jet device plays procoagulant and hemostatic effects in vivo and in vitro,and its effect is not dependent on temperature and airflow evaporation effects and is considered to be related to platelet activation,with low thermal damage to living tissue.[Chinese Medical Equipment Journal,2025,46(6):20-27]

To investigate the molecular mechanisms underlying the mechanosensitive ion channel PI-EZO2 in osteoarthritis(OA),we developed a lentiviral vector for endogenous PIEZO2 overexpression and established a stable PIEZO2-high-expressing immortalized human primary chondrocyte line.By map-ping the open reading frame of the PIEZO2 locus and designing sequence-specific sgRNA,we employed the CRISPR/Cas9 synergistic activation mediator(SAM)system to precisely integrate transcriptional ac-tivation elements into the PIEZO2 promoter region.Lentiviral-mediated targeted genomic integration en-sured endogenous PIEZO2 overexpression,confirmed by mCherry fluorescence tracing coupled with flow cytometric sorting,which revealed membrane-specific localization of PIEZO2 protein(localization effi-ciency:78.49％).Quantitative PCR demonstrated a 17-fold upregulation of PIEZO2 mRNA,while Western blotting validated enhanced membrane-localized protein expression.Strikingly,PIEZO2-overex-pressing chondrocytes exhibited hallmark OA metabolic phenotypes compared to wild-type controls:typeⅡ collagen mRNA expression decreased to 50％of baseline levels,whereas matrix metalloproteinase 13(MMP13)mRNA surged by 20-fold.These alterations recapitulated the pathological matrix metabolic phenotype observed in biomechanical OA models induced by cyclic mechanical stress(10％strain,0.5 Hz,8 h/day for 2 consecutive days).Collectively,we successfully generated a human chondrocyte model with stable PIEZO2 overexpression,which faithfully mirrors mechanotransduction-driven OA progression.This engineered cellular system provides a robust platform for dissecting PIEZO2-mediated mechanosig-naling networks and advancing targeted therapeutic discovery.

Objective To investigate the coagulation effect of a cold atmospheric plasma(CAP)jet device with helium as the working gas and to study its coagulation mechanism preliminarily.Methods A CAP jet device treatment group,a helium airflow treatment group,a hot air treatment group(60℃)and a natural coagulation group were formed according to the treatment modes of the blood samples,with 10 μL of blood samples involved in each group,in order to validate the coagulation effect of the CAP jet device in vitro;the coagulation mechanism of the CAP jet device was explored by its application to the treatment of anticoagulated whole blood,platelet-rich plasma and platelet-depleted plasma;the coagulation effect of the CAP jet device in vivo was verified with a mouse liver punctate hemorrhage model and a rabbit mesenteric hemorrhage model.Results The CAP jet device can significantly accelerate the coagulation of anticoagulated blood droplets,and the coagulation time of anticoagulated blood droplets in the CAP jet device-treated group was shortened from 28 min in the natural coagulation group to(23±1.56)s,with the difference statistically significant(P<0.05),and the CAP jet device treatment group gained advantages significantly over the helium airflow treatment group(P<0.05)and the hot air(60℃)treatment group(P<0.05)in coagulation-promoting effect;the procoagulant effect of the CAP jet device rose with the increase of platelet content in blood droplets,and the coagulation effect of platelet-rich blood droplets was significantly better than that of whole blood(P<0.05),while no coagulation was observed in platelet-poor droplets.The CAP jet device could rapidly stop hemostasis of punctate hemorrhage in mouse liver and mesenteric hemorrhage in rabbits without delayed hemorrhage occurring within 10 min,and no obvious structural abnormality of the liver and thermal damage of the tissue were found microscopically.Conclusion The CAP jet device plays procoagulant and hemostatic effects in vivo and in vitro,and its effect is not dependent on temperature and airflow evaporation effects and is considered to be related to platelet activation,with low thermal damage to living tissue.[Chinese Medical Equipment Journal,2025,46(6):20-27]

Objective To observe the expressions of nucleotide-binding oligomerization domain receptor 1(NOD1)and its mediated RIP2/NF-κB signaling pathway-related proteins and autophagy-related proteins in cerebral cortex of rats with cerebral ischemia-reperfusion injury(CIRI);To explore the possible mechanism of eye acupuncture alleviating CIRI.Methods SPF-grade male Wistar rats were randomly divided into blank control group(12 rats),sham-operation group(12 rats)and modeling group(36 rats).A CIRI model was established by improved suture method.The rats in the modeling group were randomly divided into the model group,eyeacupuncture group,outside the acupoint area group,with 12 rats in each group.Neurological deficits in rats were evaluated by Longa score,TTC staining was used to observe the cerebral infarction,HE staining was used to observe the morphology of ischemic brain tissue,electron microscopy was used to observe the formation of autophagosome in ischemic brain tissue,RT-qPCR was used to detect the mRNA expressions of NOD1,receptor interacting protein 2(RIP2),nuclear factor-κB(NF-κB)p65 in ischemic cerebral cortex,Western blot was used to detect the expressions of NOD1,RIP2,NF-κB p65 and autophagy related proteins in ischemic cerebral cortex.Results Compared with the sham-operation group,the neurological deficit score of rats in the model group significantly increased(P<0.01),the infarct volume significantly increased(P<0.01),the typical cribriform infarct foci and multiple autophagosomes appeared in the ischemic brain tissue,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly increased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly increased(P<0.01),and the protein expression of p62 significantly decreased(P<0.01).Compared with the model group,the neurological deficit score in eye acupuncture group significantly decreased(P<0.01),the cerebral infarction volume significantly decreased(P<0.01),the area of cribriform reticular infarct and the number of autophagosomes in ischemic brain tissue significantly decreased,the mRNA expressions of NOD1,RIP2 and NF-κB p65 in ischemic cerebral cortex significantly decreased(P<0.01),the protein expressions of NOD1,RIP2,p-NF-κB p65,Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5 significantly decreased(P<0.01),and the protein expression of p62 significantly increased(P<0.01).There was no statistical significance compared with outside the acupoint area group.Conclusion Eye acupuncture can attenuat the injury of neurons in CIRI rats,and its mechanism may be related to inhibiting the activation of RIP2/NF-κB signaling pathway mediated by NOD1,thereby reducing autophagy of neurons.

Total mesorectal excision (TME) is the gold standard for the treatment of middle and low rectal tumors. Since the first patient who underwent transanal total mesorectal excision (taTME) was reported in 2010, taTME has attracted wide attention from scholars at home and abroad. Fecal incontinence is one of the common complications after taTME, which severely affects the postoperative quality of life of patients. This paper systematically summarizes the commonly used assessment methods, current situation, influencing factors, treatment and nursing progress of postoperative fecal incontinence patients with taTME, in order to provide evidence for improving the quality of life of postoperative defecation incontinence patients with taTME.