Background: s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated. Methods: In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs. Results: Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment. Conclusions We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.

Background: s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated. Methods: In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs. Results: Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment. Conclusions We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.

Background: s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated. Methods: In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs. Results: Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment. Conclusions We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.

Peroxisome proliferator-activated receptor gamma(PPAR-γ)is a member of the ligand-activated nuclear tran-scription factor superfamily.Activated PPAR-γ is involved in the regulation of many central nervous system(CNS)events,and is involved in cholesterol metabolism by inducing or inhibi-ting a series of gene pathways,thereby inhibiting the deposition of β-amyloid protein(Aβ).It plays an important neuroprotec-tive role in Alzheimer's disease(AD),improves memory and cognition in AD,and is a potential target for AD.Drug develop-ment aimed at restoring cholesterol homeostasis may be a poten-tial strategy to counteract AD.By analyzing the distribution and structure of PPAR-γ,focusing on the biological correlation be-tween PPAR-γ-mediated cholesterol metabolism and AD,this paper describes the mechanism regulation of PPAR-γ on key proteins,genes and their corresponding molecules,providing a new reference for the treatment of AD.

Aim To explore the lipid-lowering effect of Dendrobium huoshanense C.Z.Tang et S.J.Cheng polysaccharides(DHP),on atherosclerotic mice and its regulatory effect on intestinal flora.Methods Twelve 8-week-old ApoE-/-mice were randomly di-vided into two groups:H-1 and H-2.Both groups were fed a high-fat diet(HFD).After eight weeks,the H-2 group received DHP(400 mg·kg-1·d-1)for six weeks,while the H-1 group received normal saline.Blood samples were collected at 0,4,8 and 14 weeks.Feces from the H-2 group were collected and labeled as B,E,F and D,and liver and kidney tissues were col-lected post-mortem at week 14.Blood indexes were de-tected using kits,and fecal flora was analyzed by met-agenomics.Results DHP significantly decreased blood lipid levels in mice(P＜0.05)without causing significant liver and kidney damage.Metagenomic a-nalysis showed that,compared with group B,the rela-tive abundance of Deferribacterota and Faecalibaculum increased in EF group,while the relative abundance of Bacteroidota,Prevotella and Lachnoclostridium de-creased(P＜0.05).Compared with EF group,the relative abundance of Bacteroidota,Prevotella and Lachnoclostridium increased in group D,while the rela-tive abundance of Deferribacterota,Bacillota,Oscilli-bacter and Faecalibaculum decreased(P＜0.05).Actinomycetota,Faecalibaculum rodentium and other bacteria increased with longer HFD feeding time(P＜0.05).Conversely,Parabacteroides and Lachnoclos-tridium decreased with increased HFD feeding time(P＜0.05).Conclusions The intestinal flora structure of mice changes significantly with the progression of atherosclerosis.DHP intervention shows a potential to improve microflora structure and lipid metabolism in mice.

A prokaryotic expression vector for the mutant diphtheria toxin CRM197 was constructed and expressed in Esch-erichia coli cells.Anti-CRM197 antibody concentrations were detected in serum samples of healthy volunteers.The crm 197 gene was codon-optimized in E.coli and cloned into the plasmid pET28a(+)under optimized expression conditions.CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography,and confirmed by western blot analysis.The puri-fied CRM197 was used to detect specific anti-CRM197 antibody levels in serum samples of different age groups.The results showed that soluble codon-optimized CRM197 was successfully expressed under optimized expression conditions.The purity of CRM197 was more than 95％,as determined with Ni-NTA spin columns and ion exchange chromatography,consistent with the single specific bands obtained by western blot analysis and detection of serum levels of the anti-CRM197 antibody.Collec-tively,these results confirmed that the proposed expression strategy achieved high-yield production of soluble CRM197,al-though high levels in human serum may affect evaluation of immune interactions with glycan-CRM197 conjugates for applica-tion as a diagnostic antigen.The diphtheria mutant toxin CRM197 is used in many conjugate vaccines.The synthetic crm 197 gene with codon optimization in pET28a was transformed into E.coli Origami B(DE3)cells.CRM197 was induced by isopro-pyl β-d-1-thiogalactopyranoside and high level accumulation of soluble CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography.The purity of the final prepara-tion reached 95％.CRM197 was used to detect the concentra-tions of the anti-CRM197 antibody in serum samples of healthy volunteers of different ages.The proposed expression strategy yielded high production of CRM197,which could interfere with evaluations of induced immune interactions by glycan-CRM197 conjugates and prohibit application as a diagnostic antigen.

The epidemiology and etiology of three suspected cases of imported skin diphtheria infection in Fujian Province were investigated.Secretion samples of patients with skin damage were collected for isolation and culture of Corynebacterium diphtheriae.Biochemical identification and mass spectrum analysis of pure cultures of suspected C.diphtheriae were conduc-ted,the virulence-related genes,including diphtheria toxin reporter(dtxR),toxin A(toxA),and toxin B(toxB)were detec-ted,and multilocus sequence typing(MLST)was performed.All three cases had typical clinical manifestations of cutaneous diphtheria,and C.diphtheriae was isolated from the damaged skin.The virulence genes of two C.diphtheriae strains isolated from two cases were identified as dtxR(+),toxA(-),and toxB(-),and the MLST type was ST-703.The virulence genes of C.diphtheriae isolated from one case were identified as dtxR(+),toxA(+),toxB(+),and the MLST type was ST-248.There is an increased risk of diphtheria in Fujian Province.C.diphtheriae without diphtheria toxin genes can also cause skin diphtheria.

Cardiovascular diseases (CVD) are the leading cause of death among residents in China and significantly impact patients' quality of life.The pathogenesis of CVD is complex,with chronic low-grade inflammation as a predominant feature.The nucleotide-binding oligomerization domain-like receptor pro-tein 3 (NLRP3) inflammasome is a key regulator of inflammatory responses and plays a crucial role in in-nate immune responses.Its abnormal activation leads to excessive inflammation and pyroptosis,which is closely associated with the development of atherosclerosis,myocardial infarction,diabetic cardiomyopa-thy,and other CVDs.Recent studies suggest that exercise,as a safe and effective non-pharmacological intervention,can mitigate cardiovascular inflammation and pyroptosis by inhibiting NLRP3 inflammasome activation,thereby playing a positive role in CVD prevention and treatment.However,the theories and mechanisms of exercise-mediated NLRP3 inflammasome intervention in CVD remain underexplored.Therefore,this review summarizes the relationship between the NLRP3 inflammasome and CVD,explores the effects of different types,intensities,and durations of exercise on NLRP3 inflammasome activation,and provides a comprehensive analysis of the signaling pathways involved in exercise interventions targe-ting the NLRP3 inflammasome for the prevention and treatment of CVD.The aim is to provide new in-sights and references for exercise-based prevention and management of CVD.

Objective:To explore the clinical characteristics,molecular characteristics,treatment and prognosis of pediatric Philadelphia chromosome-like acute lymphoblastic leukemia(Ph-like ALL)with a therapeutic target.Methods:A total of 27 patients of Ph-like ALL with targeted drug target were initially diagnosed in Children's Hospital of Soochow University from December 2017 to June 2021.The data of age,gender,white blood cell(WBC)count at initial diagnosis,genetic characteristics,molecular biological changes,chemotherapy regimen,different targeted drugs were given,and minimal residual disease(MRD)on day 19,MRD on day 46,whether hematopoietic stem cell transplantation(HSCT)were retrospective analyed,and the clinical characteristics and treatment effect were summarized.Survival analysis was performed by Kaplan-Meier method.Results:The intensity of chemotherapy was adjusted according to the MRD level during induced remission therapy in 27 patients,10 patients were treated with targeted drugs during treatment,and 3 patients were bridged with HSCT,1 patient died and 2 patients survived.Among the 24 patients who did not receive HSCT,1 patient developed relapse,and achieved complete remission(CR)after treatment with chimeric antigen receptors T cells(CAR-T).The 3-year overall survival,3-year relapse-free survival and 3-year event-free survival rate of 27 patients were(95.5±4.4)％,(95.0±4.9)％and(90.7±6.3)％respectively.Conclusion:Risk stratification chemotherapy based on MRD monitoring can improve the prognosis of Ph-like ALL in children,combined with targeted drugs can achieve complete remission as soon as possible in children whose chemotherapy response is poor,and sequential CAR-T and HSCT can significantly improve the therapeutic effect of Ph-like ALL in children whose MRD is continuously positive during induced remission therapy.

Objective:To investigate the expression of complement C3 in serums and tissues of lung adenocarcinoma patients with brain metastases and the mechanism of complement C3 in inducing epithelial mesenchymal transition (EMT).Methods:The retrospective case-control study, cell experiments and animal experiments were conducted. The serum samples from 20 healthy examinees, 20 advanced lung adenocarcinoma patients without brain metastases and 20 advanced lung adenocarcinoma patients with brain metastases at the First People's Hospital of Yancheng from January 2021 to January 2023 were collected, and the expression of complement C3 in serum samples was detected by immunoturbidimetry. At the same time, lung tissue samples were collected from 10 lung adenocarcinoma patients without brain metastases, and lung tissue and brain tissue samples were collected from 10 lung adenocarcinoma patients with brain metastases in the First People's Hospital of Yancheng. Immunohistochemistry was used to detect the expression of complement C3, C3aR, Kruppel like factor 5 (KLF5), and N-cadherin (N-cad) in the tissue samples. Using lentivirus to construct a human lung adenocarcinoma with brain metastases cell line PC14-C3 with stable overexpression of complement C3, with cells infected with empty vector virus as the control group (PC14-Ctrl). Western blotting was used to detect the expression of complement C3, KLF5, N-cad, and E-cadherin (E-cad) in PC14-C3 and PC14-Ctrl cells, and scratch assay was used to assess cell migration ability. Using the random number table method, 12 BALB/c nude mice were evenly divided into PC14-C3 group and PC14-Ctrl group. PC14-C3 cells and PC14-Ctrl cells were subcutaneously inoculated on the ventral side, and the body mass and tumor volume of the nude mice were recorded. Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of KLF5, N-cad and E-cad mRNA in various tumor cells and tumor tissues of nude mice.Results:The serum complement C3 levels in healthy individuals, lung adenocarcinoma patients without brain metastases and lung adenocarcinoma patients with brain metastases were (1.14±0.17) g/L, (1.20±0.15) g/L and (1.61±0.21) g/L, respectively. The serum complement C3 level in lung adenocarcinoma patients with brain metastases was higher than that in lung adenocarcinoma patients without brain metastases and healthy individuals, and the differences were statistically significant (both P < 0.001). The results of immunohistochemical testing showed that the proportions of positive expression areas of complement C3, C3aR, KLF5, and N-cad proteins in the lung primary lesions of lung adenocarcinoma patients with brain metastases were higher than those of patients without brain metastases, and the differences were statistically significant (all P < 0.05). The mRNA ( P < 0.05) and protein expression levels of complement C3 in PC14-C3 cells were higher than those in PC14-Ctrl cells, indicating successful transfection. The scratch assay results showed that the migration rate of PC14-Ctrl cells was (37.5±4.1)%, and the migration rate of PC14-C3 cells was (60.4±2.9)%, and the difference was statistically significant ( t = 7.86, P < 0.01). The relative expressions of EMT promoting molecules KLF5 and N-cad mRNA in PC14-C3 cells were higher than those in PC14-Ctrl cells, while the relative expression of EMT inhibiting molecule E-Cad mRNA was lower than that in PC14-Ctrl cells, and the differences were statistically significant (all P < 0.05). On the 26th day of tumor loading, the tumor volume of nude mice in PC14-C3 group was (610±10) mm 3, while that of PC14-Ctrl group was (321±30) mm 3; the body mass of nude mice in PC14-C3 group was lower than that in PC14-Ctrl group [(21.6±0.6) g vs. (23.2±0.6) g], and the differences were statistically significant (both P < 0.05). At the end of the experiment, 5 nude mice died and 1 survived in the PC14-C3 group; 1 nude mouse died and 5 survived in the PC14-Ctrl group. The relative expressions of KLF5 and N-cad mRNA in the tumor tissues of nude mice in PC14-C3 group were higher than those in PC14-Ctrl group, while the relative expression of E-Cad mRNA was lower than that in PC14-Ctrl group, and the differences were statistically significant (all P < 0.05). Conclusions:Lung adenocarcinoma patients with brain metastases have high levels of complement C3 in their serums and primary lesions. Complement C3 may induce EMT and promote the occurrence of lung adenocarcinoma brain metastases by affecting the expressions of KLF5, N-cad and E-cad.