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.

Aim To observe the effects of Wenfei Jiangzhuo formula(WFJZF)on rats with vascular de-mentia and investigate its possible mechanism of ac-tion.Methods Thirty-six healthy male SD rats were randomly divided into the sham group,model group,donepezil group,and low-dose,medium-dose and high-dose groups of Wenfei Jiangzhuo formula,with six rats per group.Except for the sham group,the other groups were prepared as VaD models,and each group was gavaged with the corresponding drugs after suc-cessful modeling,and tests were performed after three weeks of treatment.Behavioral,hippocampal CA1 area morphology,neural dendrites and mitochondrial chan-ges were observed in all groups of rats,and phospho-glycerate mutase 5(PGAM5),dynamics-related pro-teins1(Drp1),opticatrophyprotein-1(OPA1),and other proteins were detected in each group.Results Compared with the sham group,rats in the model group and each intervention group had prolonged es-cape latency(P＜0.05),a shorter number of travers-als across the platforms(P＜0.05),a sparse morphol-ogy of hippocampal neurons,a reduction in the number of secondary dendritic spines,and a rupture of the out-er membrane of the mitochondria;the expression of the PGAM5 and Drp1 proteins in hippocampal tissues was elevated(P＜0.05),and the expression of the OPA1 and Mfn1/2 protein expression decreased(P＜0.05);compared with the model group,donepezil group and Wenfei Jiangzhuo formula high-dose group of rats had shorter evasion latency(P＜0.05),increased number of times to traverse the platform(P＜0.05),increased number of hippocampal neurons,tightly packed,more secondary dendritic structures,and reduced mitochon-drial damage;the expression of PGAM5 and Drp1 pro-teins was reduced(P＜0.05),and the expression of OPA1 and Mfn1/2 proteins was elevated(P＜0.05).Conclusions Wenfei Jiangzhuo formula can regulate the PGAM5-Drp1 signaling axis to improve the balance of mitochondrial homeostasis,thus improving the cog-nitive condition of the brain and exerting cerebroprotec-tive effects.

Objective:To discuss the efficacy and safety of stereotactic electroencephalography (SEEG) in epileptogenic foci excision in pediatric patients with drug-resistant epilepsy.Methods:A total of 126 pediatric patients (<18 years old) with drug-refractory epilepsy who received SEEG-guided epileptogenic foci excision in Epilepsy Center, Aviation General Hospital from January 2015 to March 2022 were selected. The clinical data and efficacy were retrospectively analyzed, and prognoses of these pediatric patients were evaluated by Engel grading 1 year after resection.Results:(1) A total of 1289 electrodes were implanted, with a mean of (10.09±2.92) electrodes per pediatric patient; 55 pediatric patients had unilateral implant and 71 had bilateral implant. Mean EEG monitoring time was (8.69±5.71) d, ranged 3-28 d. Epileptogenic focus could be located in 114 pediatric patients (90.5%) after initial implantation under SEEG monitoring, and secondary implantation for accurate positioning was given in 12 pediatric patients (9.5%). (2) Lobectomy was performed in 27 pediatric patients (21.4%), multi-lobectomy or tailored cortical resection in 36 (28.6%), tailored cortical resection on single lobe in 60 (47.6%), and tailored cortical resections on single lobe or hippocampal amygdala resection combined with corpus callosotomy in 3 (2.4%). Minimally invasive exploring hemostasis under SEEG was performed in 13 pediatric patients (17 electrodes) and postoperative CT was normal. A little asymptomatic epidural, subdural or cerebral parenchymal hematoma spontaneously absorbed was noted in 4 pediatric patients after implantation under SEEG monitoring. No perioperative infection, CSF leakage, death or severe disability was noted. (3) Mean follow-up was performed for (26.1±7.26) months; 66 (52.3%) pediatric patients reached Engel grading I, 33 (26.2%) reached Engel grading II, 21 reached Engel grading III (16.7%), and 6 (4.8%) reached Engel grading IV. Thirteen pediatric patients with failed resection received SEEG-guided epileptogenic foci excision for the second time: 8 (76.9%) had Engel grading I and 2 had Engel grading II 1 year after follow-up, accounting for 76.9% totally.Conclusion:SEEG-guided epileptogenic foci excision is safe and effective in drug-refractory epilepsy; for pediatric patients with poor initial results, SEEG can be used to relocate the epileptogenic focus, and a second resection of epileptogenic focus can also obtain good results.

Objective To investigate the value of multi-slice spiral CT enhanced scanning combined with CT texture analysis in preoperative International Federation of Gynecology and Obstetrics(FIGO)staging of ovarian cancer.Methods A total of 126 ovarian cancer patients admitted to the Tongzhou District Maternal and Child Health Hospital and Beijing Friendship Hospital,Capital Medical University from March 2021 to September 2023 were selected as the research subjects.All patients underwent multi-slice spiral CT enhanced scanning,and their CT values were measured.Kinetics software was employed for CT texture analysis,and the texture feature-related parameters,including skewness,kurtosis,variance,entropy,and inverse difference,were calculated.The CT values and CT texture feature-related parameters among patients with different FIGO stages were compared.The diagnostic efficacy of CT enhanced scanning,CT texture analysis,and their combination in preoperative FIGO staging of ovarian cancer was evaluated by receiver operating characteristic(ROC)curve,and the consistency between the diagnosis of FIGO stage of ovarian cancer based on CT enhanced scanning,CT texture analysis,and their combination and the pathological diagnosis of FIGO stage of ovarian cancer was evaluated by Cohen's Kappa coefficient analysis.Results The CT value and entropy value of FIGO stage Ⅳ patients were significantly higher than those of FIGO stage Ⅰ,Ⅱ,and Ⅲpatients,and the CT value and entropy value of FIGO stage Ⅲ patients were significantly higher than those of FIGO stage Ⅰand Ⅱ patients(P＜0.05);there was no statistically significant difference in the CT value and entropy value between FIGO stage Ⅰ and stage Ⅱ patients(P＞0.05).There was no statistically significant difference in skewness,kurtosis,variance,and inverse difference among patients with different FIGO stages(P＞0.05).The ROC curve analysis showed that with reference to FIGO stages Ⅰ and Ⅱ,when the cut-off values of CT value and entropy value were 74.645 and 9.540,respectively,the area under the curve(AUC)of CT value and entropy value in diagnosing FIGO stage Ⅲ was 0.733 and 0.743,respectively,the specificity was 0.760 and 0.800,respectively,and the sensitivity was 0.605 and 0.674,respectively;the area under the curve(AUC)of CT value combined with entropy value in diagnosing FIGO stage Ⅲ was 0.818,the specificity was 0.820,and the sensitivity was 0.721.When the cut-off values of CT value and entropy value were 77.095 and 10.020,respectively,the AUC of CT value and entropy value in diagnosing FIGO stage Ⅳ was 0.817 and 0.797,respectively,the specificity was 0.820 and 0.820,respectively,and the sensitivity was 0.545 and 0.667,respectively;the AUC of CT value combined with entropy value in diagnosing FIGO stage Ⅳ was 0.926,the specificity was 0.900,and the sensitivity was 0.758.The consistency between CT enhanced scanning and pathology in diagnosing FIGO stage of ovarian cancer was moderate(Kappa=0.580,P＜0.05),with an accuracy rate of 72.22％(91/126);the consistency between CT texture analysis and pathology in diagnosing FIGO stage of ovarian cancer was moderate(Kappa=0.598,P＜0.05),with an accuracy rate of 73.81％(93/126);the combination of CT enhanced scanning and CT texture analysis in the diagnosis of FIGO stage of ovarian cancer had a high consistency with pathological diagnosis(Kappa=0.868,P＜0.05),with an accuracy rate of 91.27％.Conclusion Multi-slice spiral CT enhanced scanning and CT texture analysis are both reliable methods for the diagnosis of FIGO stage of ovarian cancer.The combination of CT enhanced scanning and CT texture analysis in the diagnosis of FIGO stage of ovarian cancer has a high consistency with pathological diagnosis.The combination of the two can improve the diagnostic efficiency for FIGO stage of ovarian cancer.

Objective To analyze the risk factors and survival status of Epstein-Barr virus(EBV)infection in pa-tients with aplastic anemia(AA)after haploid allogeneic hematopoietic stem cell transplantation(Haplo-HSCT).Methods Clinical data of 78 AA patients who underwent Haplo-HSCT in the hematology department of a hospital from January 1,2019 to October 31,2022 were analyzed retrospectively.The occurrence and onset time of EBV viremia,EBV-related diseases(EBV diseases),and post-transplant lymphoproliferative disorders(PTLD)were ob-served,risk factors and survival status were analyzed.Results Among the 78 patients,38 were males and 40 were females,with a median age of 33(9-56)years old;53 patients experienced EBV reactivation,with a total inci-dence of 67.9％,and the median time for EBV reactivation was 33(13,416)days after transplantation.Among pa-tients with EBV reactivation,49 cases(62.8％)were simple EBV viremia,2 cases(2.6％)were possible EBV di-seases,and 2 cases(2.6％)were already confirmed EBV diseases(PTLD).Univariate analysis showed that age 1＜40 years old at the time of transplantation,umbilical cord blood infusion,occurrence of acute graft-versus-host disease(aGVHD)after transplantation,and concurrent cytomegalovirus(CMV)infection were independent risk fac-tors for EBV reactivation in AA patients after Haplo-HSCT.Multivariate analysis showed that concurrent CMV in-fection was an independent risk factor for EBV reactivation in A A patients after Haplo-HSCT(P=0.048).Ritu-ximab intervention before stem cell reinfusion was a factor affecting the duration of EBV reactivation(P＜0.05).The mortality of EBV viremia,EBV diseases,and PTLD alone were 8.2％,50.0％,and 100％,respectively.The 2-year overall survival rate of patients with and without EBV reactivation were 85.3％,and 90.7％,respectively,difference was not statistically significant(P=0.897).However,patients treated with rituximab had 2-year lower survival rate than those who did not use it,with a statistically significant difference(P=0.046).Conclusion EBV reactivation is one of the serious complications in AA patients after Haplo-HSCT,which affects the prognosis and survival of patients.

AIM To investigate the effects of Zhuangyao Shuanglu Tongnao Formula on neuronal apoptosis in rats with cerebral ischemia-reperfusion injury based on the study of oxidative stress and inflammatory response.METHODS The rats were randomly divided into the sham operation group,the model group,the edaravone group(3.0 mg/kg),the low,medium and high dose groups(9.0,18.0,36.0 g/kg)of Zhuangyao Shuanglu Tongnao Formula,with 18 rats in each group.The middle cerebral artery occlusion/reperfusion was conducted by thread embolism method to simulate cerebral ischemia reperfusion injury in rats followed by 6 days corresponding drugs administration.Subsequently,the rats had their neurological function deficit scored by Zeal Longa scoring method;their sizes of cerebral infarction areas measured by TTC staining;their pathological damage and apoptosis of neurons in hippocampal CA1 area of ischemic penumbra of the brain tissue detected by HE staining and TUNEL staining;their SOD activity and levels of GSH,MDA,IL-6,IL-1β,TNF-α in brain tissue detected by kits;and their protein expressions of Bax,Bcl-2,caspase-3,cleaved-capase-3,TLR4,NF-κB p65,Nrf2,HO-1 in rat brain tissue determined by Western blot.RESULTS Compared with the model group,the groups intervened with edaravone,medium and high dose of Zhuangyao Shuanglu Tongnao Formula displayed improvements in the scores of nerve function defects,the rate of cerebral infarction,the rate of neuronal apoptosis,the levels of IL-6,IL-1β,TNF-α and MDA in the ischemic penumbra of brain tissues,the protein expressions of Bax and TLR4,the ratio of cleaved-capase-3/caspase-3 and p-NF-κB p65/NF-κB p65(P＜0.05),the levels of GSH,the activity of SOD and the protein expressions of Bcl-2,Nrf2 and HO-1(P＜0.05).CONCLUSION Being an inhibitor of oxidative stress and inflammatory response,Zhuangyao Shuanglu Tongnao Formula can alleviate brain injury in rats with cerebral ischemia reperfusion injury through the inhibition of neuronal apoptosis and improvement of neural function mediated by the inhibition of TLR4/NF-κB signal pathway and activation of Nrf2/HO-1 signal pathway.

Targeted regulation of hypoxia inducible factor(HIF)pathway can provide therapeutic basis for inflammatory anemia,hypoxic nephropathy,cardiovascular diseases related to chronic kidney disease and other hypoxic diseases.At present,the first drug to act on the HIF pathway,roxadustat,has been used for the treatment of renal anemia,and other prolyl hydroxylase(PHD)inhibitors are also in clinical research.This article mainly reviews the various pathways and mechanisms of the protective effect of PHD inhibitors on the kidneys.

Objective:To develop a recombinant protein vaccine based on KPC-2, a drug resistance target in Klebsiella pneumoniae, and evaluate its immunogenicity, protective efficacy and mechanism in a mouse model of pneumonia. Methods:KPC-2 was expressed in Escherichia coli and purified using GST affinity chromatography. A recombinant protein vaccine was prepared with KPC-2 and used to immunize New Zealand rabbits through subcutaneous injection. Serum samples were isolated from cardiac blood and Protein G chromatography was used to purify polyclonal antibodies against KPC-2. Opsonophagocytic killing assay was used to assess the bactericidal activity of the polyclonal antibodies in vitro. Female BALB/c mice were immunized three times with the recombinant protein vaccine, and the titers of specific IgG antibodies in serum were measured by indirect ELISA. One week after the last vaccination, the mice were infected with Klebsiella pneumoniae strain SRT through tracheal intubation, and received a single intravenous dose of meropenem (0.1 mg) 1 h later. The protective efficacy of the KPC-2 recombinant protein vaccine was evaluated by comparing the survival rates, bacterial colonization and histopathological changes between vaccine group and adjuvant group as well as the survival rates between meropenem group and normal saline group. Moreover, the protective efficacy of polyclonal antibodies against KPC-2 was evaluated through passive immunization. Results:The level of specific IgG antibodies in serum was significantly higher in the vaccine group than in the adjuvant group ( t=4.325, P<0.05). The survival rate in the vaccine group was also higher than that of the adjuvant group [70% (7/10) vs 10% (1/10), P<0.05]. Furthermore, lung inflammation was less severe and bacterial burden was reduced in the vaccine group as compared with those of the control group ( t=3.127, P<0.05). Both active and passive vaccination strategies demonstrated strong protective efficacy against Klebsiella pneumoniae infection, and had a synergistic effect when used in combination with antibiotic therapy. The polyclonal antibodies against KPC-2 had bactericidal activity in vitro ( t=5.427, P<0.05). Conclusions:The prepared KPC-2 vaccine has better immunogenicity and protective efficacy. It can induce strong humoral immune responses. This study suggest that drug resistance target may be used as a candidate antigen for future vaccine development.