Based on the previous transcriptomic experimental data of Trichinella spiralis(T.spira-lis)in this study,the larval stage specific gene TP12446 was screened and its identity in the ubiq-uitin ligase RNF family was predicted.In the study,bioinformatics methods were used to analyze its physicochemical properties and its activity to lay the foundation for further exploring the func-tion of TP12446 gene.The physicochemical properties and protein structure of TP12446 protein were predicted by bioinformatics.Its ubiquitin ligase activity was also verified by ubiquitination re-actions in vitro.The expression characteristics of TP12446 protein in different stage of T.spiralis infection were analyzed by qPCR and Western blot.Bioinformatics analysis showed that TP12446 protein was composed of 453 amino acids and its molecular weight was 51.48 kDa.The protein had a transmembrane structure and contained signal peptides.The results indicated that it was a secre-tory protein and mainly located in the cytoplasmic membrane.The protein structure analysis re-vealed that the protein contained RING and PA domain,its secondary structure was mainly com-posed of α-helix and irregular crimp and there were 10 B cell epitopes on TP12446 protein.The prediction of glycosylation and phosphorylation sites indicated that TP12446 protein contained 38 potential phosphorylation sites.Results of PPI interaction protein prediction showed that TP12446 protein had strong interaction with Usp8,Tmem37,Otub1,Otub2,Ubox5 and CD151.The results of qPCR and Western blot showed that TP12446 gene expression was the highest in the larva stage of T.spiralis,the activity of ubiquitin ligase was verified by ubiquitination reaction in vitro.TP12446 protein was a secretory hydrophobic protein with E3 ubiquitin ligase activity,which was involved in regulating cell cycle and apoptosis.

Objective:Primary pulmonary mucoepidermoid carcinoma(PMEC)is a rare malignant lung tumor that accounts for approxim-ately 0.1%-0.2%of all primary pulmonary neoplasms.Due to the non-specific clinical symptoms and epidemiological features,PMEC poses diagnostic challenges.Methods:Tissue blocks from 23 archived PMECs were collected from The First Affiliated Hospital of Soochow Uni-versity(November 2012 to December 2023).To establish definitive diagnoses,comprehensive histopathological evaluation,including histo-morphological analysis,immunohistochemistry(IHC),fluorescence in situ hybridization(FISH),and periodic acid-Schiff(PAS)staining were performed.Results:The tumors consisted of varying proportions of mucin-secreting cells(mucous cells),intermediate cells,and epidermoid cells.Immunophenotypically,CK7 was predominantly expressed in the mucous cells,whereas CK5/6,p40,and p63 were expressed in the epidermoid and intermediate cells.The Ki-67 proliferation index ranged from 5%to 60%.All tumors were negative for TTF-1 and Napsin A.Five of the tumors were positive for PD-L1(clone 22C3),with a tumor percentage score of 3%-20%.All 11 tumors tested for ALK(clone D5F3)were negative.IHC for c-Met was performed on two tumors and both were weakly positive(+).Mastermind-like transcriptional co-activator 2(MAML2)gene rearrangement was detected in 34.8%(8/23)of the tumors.Mucous cells were PAS positive.Kaplan-Meier surviv-al analysis revealed a significantly poorer prognosis for patients with lymph node metastasis,distant metastasis,advanced TNM stage(Ⅲ+Ⅳ),poor differentiation,or MAML2 gene rearrangement negativity.Univariate analysis identified poor histological differentiation,lymph node metastasis,distant metastasis,and advanced TNM stage as the major prognostic risk factors.Multivariate analysis confirmed poor differentiation and distant metastasis as independent risk factors for adverse outcomes.Conclusions:PMEC is an aggressive tumor with low incidence and non-specific clinical manifestations,leading to frequent misdiagnosis.Clinicians should maintain a high index of suspicion and ensure a thorough differential diagnosis.

OBJECTIVE To investigate the improvement effects of total flavonoids from Bidens pilosa （TFB）against Alzheimer’s disease （AD） and elucidate its potential mechanism. METHODS The network pharmacology was adopted to explore active constituents and core targets of TFB for AD， followed by gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses. Based on the results of network pharmacology， an AD model was induced in male BALB/c mice by D-galactose subcutaneous injection and aluminum chloride gavage. The effects of TFB on behavioral indicators （including escape latency， the number of platform crossings， and the proportion of dwell time spent in the original platform quadrant）， as well as on acetylcholinesterase （AChE）， acetylcholine （ACh）， choline acetyltransferase （ChAT）， amyloid β-protein （Aβ）， phosphorylated Tau protein （p-Tau）， and inflammatory factors [interleukin-1β （IL-1β）， IL-6， and tumor necrosis factor-α （TNF-α）] were investigated. Additionally， its effects on the pathological changes in hippocampal neurons， as well as the expressions of related proteins and mRNAs were evaluated. RESULTS Network pharmacology revealed 6 active components in TFB （e.g. luteolin， quercetin， kaempferol） and 165 overlapping targets with AD， including 29 core targets （Akt1， TP53， etc.）. The common targets were primarily enriched in biological processes such as positive regulation of gene expression and negative regulation of apoptotic processes， molecular functions including enzyme binding and identical protein binding， cellular components like extracellular space， plasma membrane and receptor complex， as well as signaling pathways such as cancer pathways and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. The results of animal experiments showed that， compared with model group， the pathological changes such as disordered arrangement， degeneration， and necrosis of neurons in the hippocampal CA3 region of mice in administration groups were alleviated. The escape latency （except for the low-dose TFB group）， the contents of AChE （except for the low-dose TFB group）， Aβ40， Aβ42 （except for the low-dose TFB group）， p-Tau （except for the low- and medium-dose TFB groups）， IL-1β， IL-6 （except for the low-dose TFB group）， and TNF- α in brain tissue， as well as the expressions of Bax and caspase-3 mRNA， were all significantly shortened/reduced/down-regulated. Conversely， the number of platform crossings， the proportion of dwell time spent in the original platform quadrant， the contents of ChAT and ACh， the phosphorylation levels of PI3K and Akt， and the mRNA expressions of PI3K， Akt and Bcl-2 （except for PI3K mRNA and Akt mRNA in the low- and medium-dose TFB groups， and Bcl-2 mRNA in the low-dose TFB group） were all significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS TFB can exert anti-AD effect through multiple components， multiple targets， and multiple pathways. Its underlying mechanisms may be related to the activation of the PI3K/Akt signaling pathway， improvement of the cholinergic system， reduction of Aβ deposition and Tau protein hyperphosphorylation， as well as inhibition of neuroinflammatory responses and neuronal apoptosis.

AIM:C57BL/6J mice were transplanted with fecal microbiota from a patient with maturity-onset diabetes of the young(MODY)or a healthy control.This study aimed to investigate the effects of fecal microbiota trans-plantation(FMT)on glucose and lipid metabolism,intestinal flora composition,and short-chain fatty acids(SCFAs)me-tabolism in mice,thereby providing an experimental foundation for understanding the role of gut microbiota in MODY.METHODS:A total of 36 male C57BL/6 mice were randomly allocated into three groups,including:control(CON)group(receiving PBS),normal individual-FMT(NF)group(intervened with normal human fecal microbiota)and MODY patient-FMT(MF)group(intervened with MODY patient fecal microbiota),and the mice were performed fecal microbio-ta,transplantation through oral gavage for 8 weeks.Glucose tolerance was assessed using an oral glucose tolerance test(OGTT),while insulin sensitivity was evaluated through an intraperitoneal insulin tolerance test(ITT).Blood biochemi-cal indicators were measured using ELISA,the composition of the gut microbiota was analyzed using 16S rRNA sequenc-ing,and the concentration of fecal short-chain fatty acids(SCFAs)was determined by gas chromatography-mass spec-trometry(GC-MS).RESULTS:The mice in MF group showed impaired glucose tolerance and reduced insulin sensitivi-ty.The FMT altered the composition of gut microbiota in mice,leading to significant differences in microbial diversity among groups.Specifically,the distribution of 18 bacterial species varied significantly among the three groups,with a no-table reduction in SCFA-producing bacteria observed in the MF group.Analyses of fecal SCFAs revealed that acetic acid levels were significantly lower in the MF group(P<0.05).CONCLUSION:The FMT reduces production of SCFAs by altering gut microbiota composition in mice,which was likely related to impaired glucose tolerance and decreased insulin sensitivity.

Objective:To analyze the value of abdominal CT angiography (CTA) and three-dimensional reconstruction fusion technology for larger (2 - 5 cm) gastric submucosal tumors (SMT) in the preoperative evaluation of super minimally invasive surgery(SMIS).Methods:A retrospective study was conducted, collecting data from 20 patients with gastric SMTs measuring 2 - 5 cm in diameter who were hospitalized in the the First Medical Center of the Chinese PLA General Hospital from March 2023 to December 2024. All patients underwent abdominal CTA prior to SMIS, and three-dimensional reconstructions of the tumors were performed. Clinical data were collected to analyze the visualization of the tumors and surrounding blood vessels in the fused three-dimensional images and the corresponding surgical plans.Results:In 19 cases, the abdominal CTA and three-dimensional imaging clearly and intuitively displayed the anatomical structures surrounding the tumors and provided panoramic images of small blood vessels around the tumors, enabling the selection of appropriate surgical plans. One case required conversion to laparoscopic surgery due to the intraoperative discovery of a small artery.Conclusions:The abdominal CTA examination and gastric SMT three dimensional reconstruction before SMIS can better display the anatomical location of the tumor and its relationship with surrounding small blood vessels, which is beneficial for gastroenterologists to formulate surgical plans and facilitate the smooth progress of SMIS under endoscopy.

Based on the previous transcriptomic experimental data of Trichinella spiralis(T.spira-lis)in this study,the larval stage specific gene TP12446 was screened and its identity in the ubiq-uitin ligase RNF family was predicted.In the study,bioinformatics methods were used to analyze its physicochemical properties and its activity to lay the foundation for further exploring the func-tion of TP12446 gene.The physicochemical properties and protein structure of TP12446 protein were predicted by bioinformatics.Its ubiquitin ligase activity was also verified by ubiquitination re-actions in vitro.The expression characteristics of TP12446 protein in different stage of T.spiralis infection were analyzed by qPCR and Western blot.Bioinformatics analysis showed that TP12446 protein was composed of 453 amino acids and its molecular weight was 51.48 kDa.The protein had a transmembrane structure and contained signal peptides.The results indicated that it was a secre-tory protein and mainly located in the cytoplasmic membrane.The protein structure analysis re-vealed that the protein contained RING and PA domain,its secondary structure was mainly com-posed of α-helix and irregular crimp and there were 10 B cell epitopes on TP12446 protein.The prediction of glycosylation and phosphorylation sites indicated that TP12446 protein contained 38 potential phosphorylation sites.Results of PPI interaction protein prediction showed that TP12446 protein had strong interaction with Usp8,Tmem37,Otub1,Otub2,Ubox5 and CD151.The results of qPCR and Western blot showed that TP12446 gene expression was the highest in the larva stage of T.spiralis,the activity of ubiquitin ligase was verified by ubiquitination reaction in vitro.TP12446 protein was a secretory hydrophobic protein with E3 ubiquitin ligase activity,which was involved in regulating cell cycle and apoptosis.

AIM:C57BL/6J mice were transplanted with fecal microbiota from a patient with maturity-onset diabetes of the young(MODY)or a healthy control.This study aimed to investigate the effects of fecal microbiota trans-plantation(FMT)on glucose and lipid metabolism,intestinal flora composition,and short-chain fatty acids(SCFAs)me-tabolism in mice,thereby providing an experimental foundation for understanding the role of gut microbiota in MODY.METHODS:A total of 36 male C57BL/6 mice were randomly allocated into three groups,including:control(CON)group(receiving PBS),normal individual-FMT(NF)group(intervened with normal human fecal microbiota)and MODY patient-FMT(MF)group(intervened with MODY patient fecal microbiota),and the mice were performed fecal microbio-ta,transplantation through oral gavage for 8 weeks.Glucose tolerance was assessed using an oral glucose tolerance test(OGTT),while insulin sensitivity was evaluated through an intraperitoneal insulin tolerance test(ITT).Blood biochemi-cal indicators were measured using ELISA,the composition of the gut microbiota was analyzed using 16S rRNA sequenc-ing,and the concentration of fecal short-chain fatty acids(SCFAs)was determined by gas chromatography-mass spec-trometry(GC-MS).RESULTS:The mice in MF group showed impaired glucose tolerance and reduced insulin sensitivi-ty.The FMT altered the composition of gut microbiota in mice,leading to significant differences in microbial diversity among groups.Specifically,the distribution of 18 bacterial species varied significantly among the three groups,with a no-table reduction in SCFA-producing bacteria observed in the MF group.Analyses of fecal SCFAs revealed that acetic acid levels were significantly lower in the MF group(P<0.05).CONCLUSION:The FMT reduces production of SCFAs by altering gut microbiota composition in mice,which was likely related to impaired glucose tolerance and decreased insulin sensitivity.

Objective:To analyze the value of abdominal CT angiography (CTA) and three-dimensional reconstruction fusion technology for larger (2 - 5 cm) gastric submucosal tumors (SMT) in the preoperative evaluation of super minimally invasive surgery(SMIS).Methods:A retrospective study was conducted, collecting data from 20 patients with gastric SMTs measuring 2 - 5 cm in diameter who were hospitalized in the the First Medical Center of the Chinese PLA General Hospital from March 2023 to December 2024. All patients underwent abdominal CTA prior to SMIS, and three-dimensional reconstructions of the tumors were performed. Clinical data were collected to analyze the visualization of the tumors and surrounding blood vessels in the fused three-dimensional images and the corresponding surgical plans.Results:In 19 cases, the abdominal CTA and three-dimensional imaging clearly and intuitively displayed the anatomical structures surrounding the tumors and provided panoramic images of small blood vessels around the tumors, enabling the selection of appropriate surgical plans. One case required conversion to laparoscopic surgery due to the intraoperative discovery of a small artery.Conclusions:The abdominal CTA examination and gastric SMT three dimensional reconstruction before SMIS can better display the anatomical location of the tumor and its relationship with surrounding small blood vessels, which is beneficial for gastroenterologists to formulate surgical plans and facilitate the smooth progress of SMIS under endoscopy.

Objective:Primary pulmonary mucoepidermoid carcinoma(PMEC)is a rare malignant lung tumor that accounts for approxim-ately 0.1%-0.2%of all primary pulmonary neoplasms.Due to the non-specific clinical symptoms and epidemiological features,PMEC poses diagnostic challenges.Methods:Tissue blocks from 23 archived PMECs were collected from The First Affiliated Hospital of Soochow Uni-versity(November 2012 to December 2023).To establish definitive diagnoses,comprehensive histopathological evaluation,including histo-morphological analysis,immunohistochemistry(IHC),fluorescence in situ hybridization(FISH),and periodic acid-Schiff(PAS)staining were performed.Results:The tumors consisted of varying proportions of mucin-secreting cells(mucous cells),intermediate cells,and epidermoid cells.Immunophenotypically,CK7 was predominantly expressed in the mucous cells,whereas CK5/6,p40,and p63 were expressed in the epidermoid and intermediate cells.The Ki-67 proliferation index ranged from 5%to 60%.All tumors were negative for TTF-1 and Napsin A.Five of the tumors were positive for PD-L1(clone 22C3),with a tumor percentage score of 3%-20%.All 11 tumors tested for ALK(clone D5F3)were negative.IHC for c-Met was performed on two tumors and both were weakly positive(+).Mastermind-like transcriptional co-activator 2(MAML2)gene rearrangement was detected in 34.8%(8/23)of the tumors.Mucous cells were PAS positive.Kaplan-Meier surviv-al analysis revealed a significantly poorer prognosis for patients with lymph node metastasis,distant metastasis,advanced TNM stage(Ⅲ+Ⅳ),poor differentiation,or MAML2 gene rearrangement negativity.Univariate analysis identified poor histological differentiation,lymph node metastasis,distant metastasis,and advanced TNM stage as the major prognostic risk factors.Multivariate analysis confirmed poor differentiation and distant metastasis as independent risk factors for adverse outcomes.Conclusions:PMEC is an aggressive tumor with low incidence and non-specific clinical manifestations,leading to frequent misdiagnosis.Clinicians should maintain a high index of suspicion and ensure a thorough differential diagnosis.

Objective:To investigate the clinicopathological characteristics of giant cell tumor of bone (GCTB) in children.Methods:A total of 35 cases of GCTB diagnosed at Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiaotong University School from 2016 to 2023 were collected, and a retrospective analysis of clinicopathological features and imaging findings was conducted.Results:Pediatric GCTB accounted for approximately 4.6% of total GCTB cases during the study period. There were 11 males and 24 females. The onset age ranged from 9 to 18 years (mean age 15 years, median age 16 years), with 8 cases (8/35, 22.9%) experiencing postoperative recurrence. Twenty-eight cases (28/35, 80%) primarily affected long bones, while 7 cases involved small or irregular bones. Imaging revealed osteolytic changes as the predominant feature, with 3 cases exhibited open physis, one of which had the tumor primarily at the diaphysis without crossing the physis. Histologically, pediatric GCTB resembled adult cases, characterized by mononuclear cells and osteoclast-like giant cells. Seven cases with denosumab treatment demonstrated degrees of giant cell disappearance, increased fibrous tissue and reactive bone proliferation in the stroma. One case was diagnosed as pediatric multicentric GCTB, and three cases as pediatric primary malignant GCTB, with malignant transformation into osteosarcoma. In all 35 cases, mutations in the H3F3A gene were identified, comprising 32 cases with H3.3 p.G34W mutations, one case with H3.3 p.G34V mutation, and 2 cases with H3.3 p.G34L mutations. Notably, the former two categories were successfully validated at the protein level through immunohistochemical staining, utilizing highly specific antibodies tailored for these mutation types: H3.3 p.G34W antibody and H3.3 p.G34V antibody. However, immunohistochemical staining was not available for the last category.Conclusions:Pediatric GCTB predominantly affects females and occurs primarily in long bones, mainly around the knee joint, the majority of tumors predominantly arise in the epiphysis and extend into the metaphysis; however, in cases where the epiphyseal plates are still unclosed, the tumors may be restricted to the metaphysis. Detection of H3F3A gene mutation is crucial for the diagnosis and differential diagnosis of pediatric GCTB.