OBJECTIVES: Sepsis-associated acute lung injury (S-ALI) is one of the major causes of death in intensive care unit (ICU) patients, yet its mechanisms remain incompletely understood and effective therapies are lacking. Lytic cell death of macrophages is a key driver of the inflammatory cascade in S-ALI. PANoptosis, a newly recognized form of lytic cell death characterized by PANoptosome assembly and activation, involves plasma membrane rupture (PMR) mediated by ninjurin-1 (NINJ1), a recently identified pore-forming protein. Itaconic acid is known for its anti-inflammatory effects, but its role in macrophage PANoptosis during S-ALI is unclear. This study aims to investigate the protective effect of itaconic acid on macrophage PANoptosis in S-ALI to provide new therapeutic insights. METHODS: Male specific-pathogen-free C57BL/6J mice (6-8 weeks, 18-20 g) received intraperitoneal lipopolysaccharide (LPS) to establish a classical S-ALI model. Western blotting was used to assess PANoptosome-related proteins and enzymes involved in the itaconic acid metabolic pathway, while real-time reverse transcription polymerase chain reaction and metabolomics quantified itaconic acid levels. Primary peritoneal macrophages (PMs) were pretreated with the itaconate derivative 4-octyl itaconate (4-OI) and then exposed to tumor necrosis factor alpha (TNF-α) plus interferon gamma (IFN-γ) to induce PANoptosis. Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. Western blotting was employed to quantify enzymes of the itaconate-metabolic pathway in PANoptotic macrophages, to evaluate the impact of 4-OI on PANoptosome-associated proteins, and to determine NINJ1 abundance in lung tissues from S-ALI mice and in PANoptotic macrophages. Fluorescent dye FM_4-64 was used to visualize 4-OI-mediated changes in PMR, whereas immunofluorescence staining mapped the effect of 4-OI on both the expression level and membrane localization of NINJ1 in PANoptotic macrophages. The effect of 4-OI on lactate dehydrogenase (LDH) release in culture supernatants and peripheal blood serum was assessed using a LDH assay kit, and non-denataring polyacylamide gel electrophoresis was used to assess the expression of NINJ1 in S-ALI mouse lung tissues and the impact of 4-OI on the expression of PANoptosis-associated NINJ1 multimeric reflected protein in macropahges. RESULTS: In S-ALI mouse lungs, PANoptosome components [NOD-like receptor thermal protein domain associated protein 3 (NLRP3), Gasdermin D (GSDMD), Caspase-1, Z-DNA binding protein (ZBP1), and Caspase-3] and phosphorylated mixed lineage kinase domain-like protein (MLKL) S345 were significantly upregulated (all P<0.05), while metabolomics showed compensatory increases in itaconic acid and its key enzymes [aconitate decarboxylase 1 (ACOD1)/immunoresponsive gene 1 (IRG1)]. In macrophages, 4-OI obviously suppressed PANoptosome protein expression, reduced LDH release, restored plasma membrane integrity, and inhibited NINJ1 expression and oligomerization at the membrane (P<0.05). CONCLUSIONS Itaconic acid may alleviate macrophage PANoptosis in S-ALI by inhibiting NINJ1-mediated plasma membrane rupture. Targeting NINJ1 or enhancing itaconate pathways may offer a novel therapeutic strategy for S-ALI.
Animals ; Acute Lung Injury/pathology* ; Succinates/pharmacology* ; Sepsis/complications* ; Mice, Inbred C57BL ; Male ; Mice ; Macrophages/pathology* ; Cell Membrane/metabolism* ; Lipopolysaccharides ; Hydro-Lyases

Objective To research the safety and efficacy of endoscopic rapid resection(ERR)for small gastric submucosal tumor(SMT).Method The surgical data and clinical outcomes of 133 patients with small gastric SMT underwent ERR from January 2020 to December 2023 were retrospectively analyzed.Result All the 133 patients completed endoscopic treatment in the same period.A patient underwent active full-thickness resection during operation,requiring hospitalization for conservative treatment and fasting.Small perforation occurred in seventeen patients and they were transferred to day ward for observation.115 patients completed the operation in the outpatient department,and the wound closure was reliable.Conclusion ERR is fast,safe,effective and practical for small gastric SMT.It can be used as the first choice for endoscopic therapy.

Objective To research the safety and efficacy of endoscopic rapid resection(ERR)for small gastric submucosal tumor(SMT).Method The surgical data and clinical outcomes of 133 patients with small gastric SMT underwent ERR from January 2020 to December 2023 were retrospectively analyzed.Result All the 133 patients completed endoscopic treatment in the same period.A patient underwent active full-thickness resection during operation,requiring hospitalization for conservative treatment and fasting.Small perforation occurred in seventeen patients and they were transferred to day ward for observation.115 patients completed the operation in the outpatient department,and the wound closure was reliable.Conclusion ERR is fast,safe,effective and practical for small gastric SMT.It can be used as the first choice for endoscopic therapy.

Objective:Using T2-weighted imaging(T2WI),diffusion-weighted imaging(DWI),pulsed gradient spin echo(PGSE)multiparametric magnetic resonance imaging combined with various histopathological techniques to observe the effects of reperfusion after different times of cerebral ischemia(CIRI)on the cytotoxic edema of brain tis-sues in rats.This provides a solid foundation for the establishment of ischemic stroke models and clinical diagnosis and treatment.Methods:Healthy adult male Sprague Dawley rats were randomly allocated to four groups:the sham-opera-ted(Sham)group,the ischaemia 60 min reperfusion(IR-60 min)group,the IR-120 min group,and the IR-180 min group.CIRI rat model was prepared by modified Zea Longa method.Laser doppler flowmetry combined with hematoxy-lin-eosin(HE)staining was employed to identify the model.Multiparametric magnetic resonance imaging,combined with water content of brain,immunofluorescence staining of aquaporin-4(AQP4),and Western Blot were performed to observe cytotoxic edema of brain tissues and tumour necrosis factor alpha(TNF-α),interleukin 1β(IL-1β)in brain tissue of rats in each group.Results:HE staining and determination of brain tissue water content prove that as ischemia time prolongs,the degree of cerebral cortex edema on the ischemic side increases.The T2WI results showed that the injury in the IR-60 min group began to affect the cerebral cortex,the injury in the IR-120 min group had completely affected the cerebral cortex,and the injury in the IR-180 min group was the most severe(P＜0.05),with a significant shift of the midline of the brain towards the opposite side.The relative apparent diffusion coefficient and water exchange time of the cerebral cortex on the ischemic side of the rats were found to be significantly lower than those of the sham group,with an declining trend(P＜0.05).Additionally,the values of cell membrane permeability of the cerebral cor-tex on the ischemic side of the rats were observed to be significantly higher than those of the sham group in all groups(P＜0.05).The expression of AQP4,IL-1 β and TNF-α in the cerebral cortex of ischemic side was significantly higher than that in the sham group,and showed an upward trend(P＜0.05).Conclusion:Reperfusion following 60,120 and,180 minutes of middle cerebral artery ischemia in rats can result in brain damage.Prolongation of ischemia time has been shown to exacerbate the toxic edema of brain cells and brain damage.Ischemia of 120 minutes reperfusion has been identified as the optimal modeling time for ischemic stroke.Multiparametric MRI can be utilized to monitor the mi-crostructural changes of brain tissue in rats following CIRI in vivo,providing a foundation for the visualization of early clinical diagnosis and treatment.

Objective:Using T2-weighted imaging(T2WI),diffusion-weighted imaging(DWI),pulsed gradient spin echo(PGSE)multiparametric magnetic resonance imaging combined with various histopathological techniques to observe the effects of reperfusion after different times of cerebral ischemia(CIRI)on the cytotoxic edema of brain tis-sues in rats.This provides a solid foundation for the establishment of ischemic stroke models and clinical diagnosis and treatment.Methods:Healthy adult male Sprague Dawley rats were randomly allocated to four groups:the sham-opera-ted(Sham)group,the ischaemia 60 min reperfusion(IR-60 min)group,the IR-120 min group,and the IR-180 min group.CIRI rat model was prepared by modified Zea Longa method.Laser doppler flowmetry combined with hematoxy-lin-eosin(HE)staining was employed to identify the model.Multiparametric magnetic resonance imaging,combined with water content of brain,immunofluorescence staining of aquaporin-4(AQP4),and Western Blot were performed to observe cytotoxic edema of brain tissues and tumour necrosis factor alpha(TNF-α),interleukin 1β(IL-1β)in brain tissue of rats in each group.Results:HE staining and determination of brain tissue water content prove that as ischemia time prolongs,the degree of cerebral cortex edema on the ischemic side increases.The T2WI results showed that the injury in the IR-60 min group began to affect the cerebral cortex,the injury in the IR-120 min group had completely affected the cerebral cortex,and the injury in the IR-180 min group was the most severe(P＜0.05),with a significant shift of the midline of the brain towards the opposite side.The relative apparent diffusion coefficient and water exchange time of the cerebral cortex on the ischemic side of the rats were found to be significantly lower than those of the sham group,with an declining trend(P＜0.05).Additionally,the values of cell membrane permeability of the cerebral cor-tex on the ischemic side of the rats were observed to be significantly higher than those of the sham group in all groups(P＜0.05).The expression of AQP4,IL-1 β and TNF-α in the cerebral cortex of ischemic side was significantly higher than that in the sham group,and showed an upward trend(P＜0.05).Conclusion:Reperfusion following 60,120 and,180 minutes of middle cerebral artery ischemia in rats can result in brain damage.Prolongation of ischemia time has been shown to exacerbate the toxic edema of brain cells and brain damage.Ischemia of 120 minutes reperfusion has been identified as the optimal modeling time for ischemic stroke.Multiparametric MRI can be utilized to monitor the mi-crostructural changes of brain tissue in rats following CIRI in vivo,providing a foundation for the visualization of early clinical diagnosis and treatment.

Background: Recent studies showed that the clinical outcome of moderately severe acute pancreatitis (MSAP) are different among different subgroups. Aims: To further subdivide MSAP, and explore the heterogeneity of MSAP subgroups. Methods: A retrospective analysis was performed on patients with acute pancreatitis (AP) from January 2016 to December 2020 at Northern Jiangsu People’s Hospital, including 538 patients with mild acute pancreatitis (MAP) and 461 patients with MSAP. MSAP patients were divided into four groups according to local complication and transient organ failure (TOF), including single acute peripancreatic fluid collection (APFC) without TOF group (group A), multiple APFC without TOF group (group B), other local complication without TOF group (group C) and TOF group (group D). The baseline data and the severity of AP among the four subgroups were compared. Meanwhile, the severity of disease between group A and MAP patients was also compared. Logistic regression analysis was used to evaluate the risk factors of MSAP. Results: Patients in group D were older than those in group A (P<0.05). There were statistically significant differences in different scoring systems among the four subgroups (P<0.05). The proportions of APACHE Ⅱ≥8, Glasgow≥3 and BISAP≥3 in group D were significantly higher than those in the other three groups (P<0.05). There were significant differences in levels of Ca

Objective To investigate the guidance of the endoscopic ultrasonography for the surgery selec-tion of patients with low rectal cancerthrough analyzing the accuracy of endoscopic ultrasonography in preoperative TN staging. Methods Eighty-seven cases with low rectal cancer received preoperative endoscopic ultrasonography examination,the preoperative staging and the postoperative pathologic comparison. The EUS accuracy of preopera-tive staging of rectal cancer was evaluated. Results The preoperative staging with endoscopic ultrasonography for patients with low rectal cancer,100% in T1 stage,96.0% in T2 stage,85.7% inT3 stage,and 100% in T4 stage. The preoperative staging and the postoperative pathologic comparison in T stage were consistent(Kappa = 0.903, P < 0.05). The preoperative staging with endoscopic ultrasonography for patients with low rectal cancer ,87.0% in N0 stage,78.6% in N1 stage,and 100% in N2. The preoperative and the postoperative pathologic comparisons in N stage were consistent(Kappa = 0.768,P < 0.05). Conclusion The endoscopic ultrasonography had a certain advantage in the clinical preoperative evaluation for patients with low rectal cancer ,especially for invasion depth and the judgment of lymph node metastasis ,with a higher accuracy.

Objective]To discuss the effect of sciatic nerve injury on the expressions of tumor necrosis factor-alpha(TNF-α), interleukin-1β(IL-1β)and interleukin-10(IL-10)in anterior cingulate cortex(ACC),and further to explain their roles resided in the development of neuropathic pain.[Method]With use of the methods of behavioral test,western blot and immunohistochemistry, we examine the effects of spared sciatic nerve injury(SNI)on the expressions of TNF-α,IL-1β,and IL-10 in ACC,and observe the effect of the neutralizing antibody of TNF-α,IL-1β on the rat mechanical allodynia.[Result]In present experiment ,SNI increased the protein levels of TNF-α,IL-10,but not IL-1β in ACC. Increased TNF-α-IR and IL-10-IR in ACC is located in neurons ,but not astrocytes and microglia at 7 d following L5-VRT. Pre-treatment with anti-TNFα antibody but not anti-IL-1βantibody into ACC significantly increased the rat paw withdrawal threshold to von Frey hairs.[Conclusion]These data suggested that the increased TNF-αin ACC neurons might be responsible for the development of neuropathic pain.

Objective To evaluate the application of sentinel lymph node biopsy in surgical operation for clinical stage N0 differentiated thyroid carcinoma.Methods One hundred and six patients who were diagnosed clinical stage N0 differentiated thyroid carcinoma in Department of Mammary Gland Thyroid Surgery of Qingdao Women and Children's Hospital between Jan.2012 and Jan.2013 were reviewed retrospectively.Carbon nanoparticles were used to trace sentinel lymph node in the surgery,the patients were divided into the dissection group (46 cases included 2 cases whose sentinel lymph nodes were not detected) and the non dissection group (60 cases) based on the rapid pathological results.The operation index,complications incidence and short-term outcomes were compared.Result The operative time,intraoperative blood loss and the faulty excision of parathyroid gland in dissection group is much higher than those in non dissection group (P ＜0.05).The incidence of provisionality hypoparathyroidism in dissection group is much higher than those in non dissection group (P ＜ 0.05).There were no permanent hypoparathyroidism and damage of laryngeal nerve in the two groups.Postoperative outpatient follow up was 30 months,no significant differences were seen in local regional recurrence rates(6.5％ vs 10％) between the two groups (P ＞ 0.05).There were no distant metastasis occurred in the two groups.Conclusions Lymph node metastasis can be found by sentinel lymph node biopsy,it is helpful for us to decide whether dissect central lymph node or not.Carbon nanoparticles is safe,which is helpful to protect the parathyroid gland.

Objective:To investigate the roles and regulation mechanism of miR-31 in human cutaneous squamous cell carcino-ma (cSCC) growth. Methods:cSCC cells were transfected with the antisense oligonucleotide (ASO) of miR-31, and the cSCC growth was tested by colony formation and in vivo tumor formation assays. The target gene of miR-31 was validated by Western blot and green fluorescent protein (GFP) reporter assay. The cells were then transfected with the siRNA of the target gene, and the effect of the target gene on cell growth was preformed by colony formation assay. Finally, real-time PCR and immunohistochemistry were used for analy-sis of the expression of miR-31 and its target gene. Results:miR-31 ASO resulted in a low number of cell colonies and small tumor vol-ume (P<0.05). Western blot showed that the cells with miR-31 ASO had a higher protein level of large tumor suppressor homolog 2 (LATS2) than the control. The 3' UTR of LATS2 had a binding site with miR-31, and miR-31 ASO increased the GFP intensity con-trolled by LATS2 3' UTR, whereas no effect was observed on the mutant LATS2 3' UTR. Western blot showed that LATS2 siRNA inhib-ited the expression of LATS2 protein by about 80%. Knocking down of LATS2 increased the colony number by about 70%or 1.3-fold in cSCC cells. Real-time PCR showed that miR-31 was overexpressed in most cSCC tissues, compared with normal tissues. An inverse relationship existed between miR-31 and LATS2 expression levels. Immunohistochemistry validated that LATS2 was downregulated in cSCC tissues. Conclusion:miR-31, which functions as an oncogene, promotes cSCC growth by suppressing LATS2 expression. Our da-ta suggest that miR-31 is a potential miRNA-based therapeutic target for cSCC growth.