Objective:To investigate the spatial distribution patterns of local tumor progression（LTP）after microwave ablation（MWA）for colorectal liver metastases（CRLMs）and identify high-risk progression zones.Methods:A total of 471 CRLM lesions from 246 patients treated with MWA at the Chinese PLA General Hospital between September 2009 and March 2022 were retrospectively analyzed. Three-dimensional visualized MRI image fusion technology was employed to evaluate the spatial relationship between ablation margins（AM）and LTP. The liver was partitioned into nine specific anatomical regions. Machine learning（Boruta algorithm）was used to assess the importance of these regions on LTP risk. Multivariate analysis of LTP was performed at the tumor level and at the patient level using the Cox mixed effects model and the Cox regression model，respectively.Results:LTP occurred in 115 lesions，with an LTP rate of 40.0%（80/200）in ablated lesions which were located in the high-risk progression area，and 12.9%（35/271）in low-risk progression area. Multivariate analysis identified AM < 5 mm，tumor size ≥3 cm and location in high-risk zones as independent risk factors for LTP. Notably，LTP in high-risk zones predominantly clustered around the ablation needle tip.Conclusions:Post-MWA LTP in CRLMs exhibits distinct spatial clustering，particularly at the needle tip within high-risk progression zones. These findings provide critical insights for optimizing ablation strategies and improving clinical outcomes.

Objective To investigate the influencing factors of disease progression and prognosis in patients with large artery atherosclerotic(LAA)cerebral infarction and analyze the value of plasma cyclin-dependent kinase 9(CDK9)level in the diagnosis and treatment of LAA cerebral infarction.Methods Patients with acute cerebral infarction admitted to the Department of Neurology of the Affili-ated Hospital of Yangzhou University between March 1,2022,and November 20,2023,were select-ed.According to the diagnostic criteria,98 patients with acute LAA(LAA group)and 33 patients with acute small artery occlusion(SAO)cerebral infarction(SAO group)were selected.Additionally,40 healthy individuals matched for age and gender from the Health Examination Center were included as control group.Based on whether the condition of LAA cerebral infarction patients progressing,they were divided into progressive cerebral infarction(PCI)group(39 patients)and the non-pro-gressive cerebral infarction(NPCI)group(59 patients).During the 3-month follow-up period,6 patients from the 98 LAA cerebral infarction patients were lost.According to the modified Rankin Scale(mRS)score at 90 days of follow-up,patients were divided into good prognosis group(mRS score≤2,59 patients)and poor prognosis group(mRS score＞2,33 patients).Fasting lipid in-dices[total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),glucose(GLU),glycated hemoglobin(HbA1c),and homocysteine(Hey)]were collected on the second day after admission.The National Institutes of Health Stroke Scale(NIHSS)was used to assess the degree of neurological impairment in cerebral infarction patients.Enzyme-linked immunosorbent assay(ELISA)was used to measure plasma CDK9 levels in different groups;factors influencing disease progression in patients with acute LAA cerebral infarction were explored;and receiver operating characteristic curves were plotted to evalu-ate the predictive value of CDK9 in patients with acute LAA cerebral infarction.Results Compared with the control group,the LAA group had lower HDL-C level and higher CDK9 level(P＜0.05).The LAA group had a higher proportion of diabetes history,larger infarction volume,higher NIHSS score at admission,and higher CDK9 level compared with the SAO group(P＜0.05).Binary Logistic regression analysis showed that diabetes history and plasma CDK9 levels were influencing factors for LAA cerebral infarction.There were statistically significant differences in the proportion of diabetes history,HbA1c,random GLU,and CDK9 levels between the NPCI and PCI groups(P＜0.05).Diabetes history and plasma CDK9 levels were influencing factors for disease progression in patients with acute LAA cerebral infarction.The area under the ROC curve for CDK9 in predicting acute LAA cerebral infarction was 0.854 5(95％CI,0.794 1 to 0.9148).When the CDK9 level was 602.1 ng/L,the Youden index was maximum(0.604),with a corresponding sensitivity of 0.849 and specificity of 0.755.The NIHSS score,infarction volume,and plasma CDK9 level were higher in the poor prognosis group compared with the good prognosis group(P＜0.01).Correlation analysis showed a positive correlation between mRS scores and CDK9 levels(r=0.485,P＜0.01).Conclusion Plasma CDK9 levels are significantly elevated,and is an influencing factor.It is positively correlated with disease progression and poor prognosis in acute cerebral infarction and has certain predictive value for the progression of LAA cerebral infarction.

Objective:To investigate the clinical value of puncture biopsy for the diagnosis of vessels that encapsulate tumor clusters (VETC) and macrotrabecular-massive (MTM) subtypes of hepatocellular carcinoma (HCC).Methods:One hundred and eighty-four patients with HCC who underwent surgical resection at the Fifth Medical Centre of Chinses PLA General Hospital from November 2023 to July 2024 were prospectively collected, including 154 males and 30 females, aged (57.1±8.6) years. By simulating the clinical puncture procedure, puncture biopsy tissue specimens were obtained postoperatively from the patient's isolated tumors. The puncture biopsies and surgical resection specimens were stained with HE and CD34, and evaluated for VETC and MTM. Patients were divided into two groups based on the histopathological VETC results of surgically resected specimens: the VETC-positive group ( n=41) and the VETC-negative group ( n=143); and two groups based on the histopathological MTM results of surgically resected specimens: the MTM-positive group ( n=39) and the MTM-negative group ( n=145). Clinical data such as gender, age, tumor length, and alpha-fetoprotein (AFP) were recorded. Logistic regression analysis was performed to screen the risk factors of VETC and MTM. Evaluating the diagnostic efficacy of puncture biopsy for VETC and MTM. Results:The results of multivariable logistic analysis showed that puncture biopsy VETC-positive ( OR=63.97, 95% CI: 16.28-251.29), grade of M2 microvascular invasion ( OR=5.07, 95% CI: 1.31-19.59) and tumor length ≥5 cm ( OR=3.42, 95% CI: 1.11-10.52) were the risk factors for VETC-positive (all P<0.05); whereas the risk factors for MTM-positive were only puncture biopsy MTM-positive ( OR=34.78, 95% CI: 12.06-100.29, P<0.001). Puncture biopsy correctly diagnosed VETC subtype in 163 patients with a diagnostic accuracy of 0.89, sensitivity of 0.61, specificity of 0.97, positive predictive value (PPV) of 0.83, and negative predictive value (NPV) of 0.90; MTM subtype was correctly diagnosed in 164 patients with a diagnostic accuracy of 0.89, sensitivity of 0.72, specificity of 0.94, PPV of 0.76, and NPV of 0.93. Using the three indicators of puncture biopsy diagnosis, tumor length and AFP level as a combined indicator, the accuracy to diagnose VETC was 0.83, sensitivity was 0.71, specificity was 0.87, PPV was 0.60, and NPV was 0.91; and the combined indicator diagnosis of MTM had a diagnostic accuracy of 0.85, a sensitivity of 0.82, specificity of 0.86, PPV of 0.68 and NPV of 0.95. Conclusion:Puncture biopsy has high specificity and accuracy in the diagnosis of VETC and MTM subtypes, but the sensitivity is relatively limited, and the role of puncture combined with clinical factors in improving diagnostic efficacy is limited.

Objective To investigate the role of Glycoprotein non-metastatic melanoma protein B(GPNMB)in hypoxia-induced epithelial-mesenchymal transition(EMT)in human chorionic trophoblast HTR-8/SVneo cells.Methods HTR-8/SVneo cells were cultured in vitro to investigate the effect of hypoxia on GPNMB expression.The cells were transfected with either a GPNMB overexpression plasmid(pcDNA3.1-GPNMB),small interfering RNA targeting GPNMB(si-GPNMB-1/2),or their respective negative controls(pcDNA3.1-NC or si-NC),and were also treated with the autophagy agonist rapamycin(Rap).The experimental groups were categorized as follows:Normoxia,Hypoxia,Normoxia/Hypoxia+si-NC or si-GPNMB,Normoxia/Hypoxia+pcDNA3.1-NC or pcDNA3.1-GPNMB,Normoxia/Hypoxia+Rap,and Hypoxia+Rap+pcDNA3.1-NC or pcDNA3.1-GPNMB.GPNMB expression levels were evaluated using qRT-PCR,Western blotting,and immunofluorescence staining.The expression of autophagy-related proteins(LC3B Ⅱ/Ⅰ,p62)and epithelial-mesenchymal transition(EMT)markers(E-cadherin,N-cadherin)was analyzed by Western blotting.Cell migration and invasion capacities were assessed using wound healing and Transwell assays.Results Compared with the Normoxia group,the mRNA and protein levels of GPNMB were downregulated in the Hypoxia group.Additionally,the protein levels of p62 and N-cadherin were reduced,while LC3B Ⅱ/Ⅰ and E-cadherin expression levels were increased(P<0.05).Compared with the Hypoxia+si-NC group,the Hypoxia+si-GPNMB-2 group showed significantly decreased protein levels of p62 and N-cadherin,along with elevated levels of LC3B Ⅱ/Ⅰ and E-cadherin(P<0.05).Compared with the Hypoxia+pcDNA3.1-NC group,the Hypoxia+pcDNA3.1-GPNMB group exhibited opposite trends.Notably,compared with the Hypoxia group,the Hypoxia+Rap group showed increased LC3B Ⅱ/Ⅰ and E-cadherin levels,accompanied by reduced p62 and N-cadherin levels(P<0.05).However,compared with the Hypoxia+pcDNA3.1-GPNMB group,the Hypoxia+Rap+pcDNA3.1-GPNMB group attenuated the promoting effect of GPNMB overexpression on EMT in HTR-8/SVneo cells,as evidenced by decreased p62 and N-cadherin protein expression levels and increased LC3BⅡ/Ⅰ and E-cadherin protein expression levels(P<0.05).Conclusion In hypoxia-induced HTR-8/SVneo cells,GPNMB inhibits autophagy,promotes the epithelial-mesenchymal transition,and enhances cell migration and invasion.

Objective To investigate the role of Glycoprotein non-metastatic melanoma protein B(GPNMB)in hypoxia-induced epithelial-mesenchymal transition(EMT)in human chorionic trophoblast HTR-8/SVneo cells.Methods HTR-8/SVneo cells were cultured in vitro to investigate the effect of hypoxia on GPNMB expression.The cells were transfected with either a GPNMB overexpression plasmid(pcDNA3.1-GPNMB),small interfering RNA targeting GPNMB(si-GPNMB-1/2),or their respective negative controls(pcDNA3.1-NC or si-NC),and were also treated with the autophagy agonist rapamycin(Rap).The experimental groups were categorized as follows:Normoxia,Hypoxia,Normoxia/Hypoxia+si-NC or si-GPNMB,Normoxia/Hypoxia+pcDNA3.1-NC or pcDNA3.1-GPNMB,Normoxia/Hypoxia+Rap,and Hypoxia+Rap+pcDNA3.1-NC or pcDNA3.1-GPNMB.GPNMB expression levels were evaluated using qRT-PCR,Western blotting,and immunofluorescence staining.The expression of autophagy-related proteins(LC3B Ⅱ/Ⅰ,p62)and epithelial-mesenchymal transition(EMT)markers(E-cadherin,N-cadherin)was analyzed by Western blotting.Cell migration and invasion capacities were assessed using wound healing and Transwell assays.Results Compared with the Normoxia group,the mRNA and protein levels of GPNMB were downregulated in the Hypoxia group.Additionally,the protein levels of p62 and N-cadherin were reduced,while LC3B Ⅱ/Ⅰ and E-cadherin expression levels were increased(P<0.05).Compared with the Hypoxia+si-NC group,the Hypoxia+si-GPNMB-2 group showed significantly decreased protein levels of p62 and N-cadherin,along with elevated levels of LC3B Ⅱ/Ⅰ and E-cadherin(P<0.05).Compared with the Hypoxia+pcDNA3.1-NC group,the Hypoxia+pcDNA3.1-GPNMB group exhibited opposite trends.Notably,compared with the Hypoxia group,the Hypoxia+Rap group showed increased LC3B Ⅱ/Ⅰ and E-cadherin levels,accompanied by reduced p62 and N-cadherin levels(P<0.05).However,compared with the Hypoxia+pcDNA3.1-GPNMB group,the Hypoxia+Rap+pcDNA3.1-GPNMB group attenuated the promoting effect of GPNMB overexpression on EMT in HTR-8/SVneo cells,as evidenced by decreased p62 and N-cadherin protein expression levels and increased LC3BⅡ/Ⅰ and E-cadherin protein expression levels(P<0.05).Conclusion In hypoxia-induced HTR-8/SVneo cells,GPNMB inhibits autophagy,promotes the epithelial-mesenchymal transition,and enhances cell migration and invasion.

Objective:To investigate the clinical value of puncture biopsy for the diagnosis of vessels that encapsulate tumor clusters (VETC) and macrotrabecular-massive (MTM) subtypes of hepatocellular carcinoma (HCC).Methods:One hundred and eighty-four patients with HCC who underwent surgical resection at the Fifth Medical Centre of Chinses PLA General Hospital from November 2023 to July 2024 were prospectively collected, including 154 males and 30 females, aged (57.1±8.6) years. By simulating the clinical puncture procedure, puncture biopsy tissue specimens were obtained postoperatively from the patient's isolated tumors. The puncture biopsies and surgical resection specimens were stained with HE and CD34, and evaluated for VETC and MTM. Patients were divided into two groups based on the histopathological VETC results of surgically resected specimens: the VETC-positive group ( n=41) and the VETC-negative group ( n=143); and two groups based on the histopathological MTM results of surgically resected specimens: the MTM-positive group ( n=39) and the MTM-negative group ( n=145). Clinical data such as gender, age, tumor length, and alpha-fetoprotein (AFP) were recorded. Logistic regression analysis was performed to screen the risk factors of VETC and MTM. Evaluating the diagnostic efficacy of puncture biopsy for VETC and MTM. Results:The results of multivariable logistic analysis showed that puncture biopsy VETC-positive ( OR=63.97, 95% CI: 16.28-251.29), grade of M2 microvascular invasion ( OR=5.07, 95% CI: 1.31-19.59) and tumor length ≥5 cm ( OR=3.42, 95% CI: 1.11-10.52) were the risk factors for VETC-positive (all P<0.05); whereas the risk factors for MTM-positive were only puncture biopsy MTM-positive ( OR=34.78, 95% CI: 12.06-100.29, P<0.001). Puncture biopsy correctly diagnosed VETC subtype in 163 patients with a diagnostic accuracy of 0.89, sensitivity of 0.61, specificity of 0.97, positive predictive value (PPV) of 0.83, and negative predictive value (NPV) of 0.90; MTM subtype was correctly diagnosed in 164 patients with a diagnostic accuracy of 0.89, sensitivity of 0.72, specificity of 0.94, PPV of 0.76, and NPV of 0.93. Using the three indicators of puncture biopsy diagnosis, tumor length and AFP level as a combined indicator, the accuracy to diagnose VETC was 0.83, sensitivity was 0.71, specificity was 0.87, PPV was 0.60, and NPV was 0.91; and the combined indicator diagnosis of MTM had a diagnostic accuracy of 0.85, a sensitivity of 0.82, specificity of 0.86, PPV of 0.68 and NPV of 0.95. Conclusion:Puncture biopsy has high specificity and accuracy in the diagnosis of VETC and MTM subtypes, but the sensitivity is relatively limited, and the role of puncture combined with clinical factors in improving diagnostic efficacy is limited.

Objective:To investigate the spatial distribution patterns of local tumor progression（LTP）after microwave ablation（MWA）for colorectal liver metastases（CRLMs）and identify high-risk progression zones.Methods:A total of 471 CRLM lesions from 246 patients treated with MWA at the Chinese PLA General Hospital between September 2009 and March 2022 were retrospectively analyzed. Three-dimensional visualized MRI image fusion technology was employed to evaluate the spatial relationship between ablation margins（AM）and LTP. The liver was partitioned into nine specific anatomical regions. Machine learning（Boruta algorithm）was used to assess the importance of these regions on LTP risk. Multivariate analysis of LTP was performed at the tumor level and at the patient level using the Cox mixed effects model and the Cox regression model，respectively.Results:LTP occurred in 115 lesions，with an LTP rate of 40.0%（80/200）in ablated lesions which were located in the high-risk progression area，and 12.9%（35/271）in low-risk progression area. Multivariate analysis identified AM < 5 mm，tumor size ≥3 cm and location in high-risk zones as independent risk factors for LTP. Notably，LTP in high-risk zones predominantly clustered around the ablation needle tip.Conclusions:Post-MWA LTP in CRLMs exhibits distinct spatial clustering，particularly at the needle tip within high-risk progression zones. These findings provide critical insights for optimizing ablation strategies and improving clinical outcomes.

Objective To investigate the role and potential mechanisms of tumor necrosis factor alpha-inducible protein 8-like molecule 1(TNFAIP8L1)in acute liver injury in mice.Methods The second generation of C57BL/6J male wild-type(WT)mice and the C57BL/6J female TNFAIP8L1+/-mice and WT mice were selected to further self-breed the third generation of male TNFAIP8L1-/-mice and the third generation of WT male mice.Five normal third-generation male WT mice and five normal third-generation male TNFAIP8L1-/-mice were selected.The serum alanine aminotransferase(ALT)levels of the two types of normal mice were measured and compared.The infiltration of inflammatory cells and cell necrosis in the liver tissues of the two types of normal mice were observed after hematoxylin & eosin(HE)staining.Flow cytometry was used to detect the percentages of neutrophils(Neu),eosinophils(EOS),dendritic cells(DC),bone marrow-derived macrophages(BMDMs),and bone marrow-derived mononuclear cell(BMNCs)in the liver myeloid cell subsets of the two types of normal mice.Another 5 third-generation male WT mice and 4 third-generation male TNFAIP8L1-/-mice were selected to induce acute liver injury mouse models using lipopolysaccharide(LPS)/D-galactosamine(D-Gal).After 24 hours,the serum ALT levels of the two types of acute liver injury mice were detected and compared,the infiltration of inflammatory cells and cell necrosis in the liver tissues of the two types of acute liver injury mice were observed,and the percentages of Neu,EOS,DC,BMDMs and BMNCs in the liver myeloid cell subsets of the two types of acute liver injury mice were measured by using the above methods.Results There was no significant difference in the percentages of Neu,EOS,DC,BMDMs and BMNCs,and serum ALT levels in the livermyeloid cell subsets of normal WT mice and TNFAIP8L1-/-mice(P＞0.05).HE staining results of liver tissues in normal WT mice and TNFAIP8L1/mice showed that hepatic lobules were structurally complete and clear,hepatocytes were morphologically normal and arranged neatly,and there was no obvious inflammatory cell infiltration or cell necrosis.Twenty-four hours after acute liver injury,the percentages of Neu and BMNCs in the liver myeloid cell subsets and the serum ALT levels in the liver tissues of TNFAIP8L1-/-mice were significantly higher than those of WT mice(P＜0.05);there was no significant difference in the percentages of EOS,DC and BMDMs in the liver myeloid cell subsets of mice between the two groups(P＞0.05).In the liver tissues of WT mice with acute liver injury,hepatic lobules were structurally blurred,hepatocytes were swollen with scattered vacuolated steatosis,and a small amount of inflammatory cells were infiltrated.In the liver tissues of TNFAIP8L1/mice with acute liver injury,hepatic lobules were structurally non-existent,and hepatocytes were severely damaged and extensively necrotic,with a large amount of inflammatory cell infiltration.Conclusion The deficiency of the TNFAIP8L1 gene in mice does not affect the development of liver myeloid cells and the homeostasis of the liver.TNFAIP8L1 plays an inhibitory role in the occurrence and development of acute liver injury.TNFAIP8L1 gene deficiency aggravates LPS/D-Gal-induced acute liver injury,possibly by increasing Neu and BMNCs infiltration and recruiting other types of immune cells to infiltrate liver tissues,thereby exacerbating liver cell necrosis.

[This corrects the article DOI: 10.1016/j.apsb.2022.08.024.].