Objective:To study the effect of inhibiting heat shock protein 70 ( HSP70) gene expression on the proliferation, invasion and migration of cholangiocarcinoma cells and its mechanism. Methods:Tumor tissues and adjacent normal tissue samples from 23 patients with cholangiocarcinoma who underwent surgery in the Hunan Second People′s Hospital from January 2022 to June 2023 were collected. The mRNA and protein expressions of HSP70 in cholangiocarcinoma tissues, human cholangiocarcinoma cells (HuCC-T1), normal bile duct tissues and human intrahepatic biliary epithelial cells (HIBEpiC) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. HuCC-T1 cells were cultured in vitro, and a HuCC-T1 cell line with stably knocked down HSP70 gene (HuCC-T1-HSP70-KD group) was obtained by screening after infection with shRNA lentivirus. Cell counting kit-8 (CCK-8) assay and Transwell assay were used to detect the effects of inhibiting HSP70 gene expression on the proliferation, invasion and migration abilities of HuCC-T1 cells. Western blot was used to detect the protein expressions of Toll-like receptor (TLR) 4, phosphorylated extracellular regulated protein kinases 1/2 (p-ERK1/2), ERK1/2, β-catenin, c-myc, Snail and E-cadherin after inhibiting HSP70 gene expression in HuCC-T1 cells. Results:Compared with normal bile duct tissues and HIBEpiC cells, the mRNA and protein expressions of HSP70 in cholangiocarcinoma tissues and HuCC-T1 cells were significantly higher (all P<0.05). After inhibiting HSP70 gene expression in HuCC-T1 cells, the proliferation, invasion and migration abilities of cells in the HuCC-T1-HSP70-KD group were significantly decreased (all P<0.05); the protein expressions of TLR4, p-ERK1/2, β-catenin, c-myc and Snail in the HuCC-T1-HSP70-KD group were significantly decreased, while the protein expression of E-cadherin was significantly increased (all P<0.05). Conclusions:Silencing HSP70 gene expression can significantly inhibit the proliferation, invasion and migration abilities of cholangiocarcinoma cells. The mechanism may be that after the down-regulation of HSP70 gene expression, its activation of downstream TLR4 and MAPK pathways is significantly inhibited, thereby affecting the proliferation, invasion and migration of cholangiocarcinoma cells.

Objective:To study the effect of inhibiting heat shock protein 70 ( HSP70) gene expression on the proliferation, invasion and migration of cholangiocarcinoma cells and its mechanism. Methods:Tumor tissues and adjacent normal tissue samples from 23 patients with cholangiocarcinoma who underwent surgery in the Hunan Second People′s Hospital from January 2022 to June 2023 were collected. The mRNA and protein expressions of HSP70 in cholangiocarcinoma tissues, human cholangiocarcinoma cells (HuCC-T1), normal bile duct tissues and human intrahepatic biliary epithelial cells (HIBEpiC) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. HuCC-T1 cells were cultured in vitro, and a HuCC-T1 cell line with stably knocked down HSP70 gene (HuCC-T1-HSP70-KD group) was obtained by screening after infection with shRNA lentivirus. Cell counting kit-8 (CCK-8) assay and Transwell assay were used to detect the effects of inhibiting HSP70 gene expression on the proliferation, invasion and migration abilities of HuCC-T1 cells. Western blot was used to detect the protein expressions of Toll-like receptor (TLR) 4, phosphorylated extracellular regulated protein kinases 1/2 (p-ERK1/2), ERK1/2, β-catenin, c-myc, Snail and E-cadherin after inhibiting HSP70 gene expression in HuCC-T1 cells. Results:Compared with normal bile duct tissues and HIBEpiC cells, the mRNA and protein expressions of HSP70 in cholangiocarcinoma tissues and HuCC-T1 cells were significantly higher (all P<0.05). After inhibiting HSP70 gene expression in HuCC-T1 cells, the proliferation, invasion and migration abilities of cells in the HuCC-T1-HSP70-KD group were significantly decreased (all P<0.05); the protein expressions of TLR4, p-ERK1/2, β-catenin, c-myc and Snail in the HuCC-T1-HSP70-KD group were significantly decreased, while the protein expression of E-cadherin was significantly increased (all P<0.05). Conclusions:Silencing HSP70 gene expression can significantly inhibit the proliferation, invasion and migration abilities of cholangiocarcinoma cells. The mechanism may be that after the down-regulation of HSP70 gene expression, its activation of downstream TLR4 and MAPK pathways is significantly inhibited, thereby affecting the proliferation, invasion and migration of cholangiocarcinoma cells.

Background and Aims:Kaempferol is a natural flavonoid compound that can regulate various processes and activities associated with cancer,such as the cell cycle,oxidative stress,apoptosis,proliferation,metastasis,and angiogenesis. Its potential for treating breast cancer has been validated in some studies,but its mechanism of action remains unclear. Therefore,this study was conducted to explore the potential targets and pathways of kaempferol in the treatment of breast cancer and establish a prognostic model.Methods:Using network pharmacology and bioinformatics approaches,the intersection targets of kaempferol for the treatment of breast cancer were obtained from databases such as HERB,GeneCards,STRING,PubChem,RCSB PDB,and TCGA. Protein interaction,GO analysis,and KEGG pathway enrichment analysis were performed,followed by molecular docking for validation. Prognostic-related genes were identified through LASSO-Cox regression analysis to establish a risk score model. The prognostic analysis for each gene,and the correlation between risk score and immune infiltration were analyzed. Results:Network pharmacology analysis identified 55 intersection targets. Protein interaction analysis revealed five potential key genes (Akt1,Bcl2,CASP3,ESR1,AR). GO analysis showed that kaempferol treatment of breast cancer involved 1604 biological process entries,20 cellular component entries,and 121 molecular function entries. KEGG pathway enrichment mainly included pathways related to chemical carcinogenesis,reactive oxygen species signaling,AGE-RAGE signaling in diabetic complications,TNF signaling,and IL-17 signaling. Molecular docking results indicated that kaempferol had a strong affinity for key targets,with the best binding effect observed with ESR1 (-9.1 kcal/mol). Ten prognostic genes (Bcl2,CYP1B1,DPP4,GSTM1,GSTM2,MMP1,NCOA2,NOS2,NR1I3,PTGS2) were obtained through LASSO-Cox regression analysis,and a risk score model was established. The AUC of this model for predicting breast cancer patient prognosis was greater than 0.5. Single-gene survival analysis indicated that higher expression of Bcl2 (HR=0.61,95％ CI=0.43-0.86,P=0.005),CYP1B1 (HR=0.68,95％ CI=0.49-0.94,P=0.022),GSTM1 (HR=0.68,95％ CI=0.47-0.98,P=0.037),GSTM2 (HR=0.64,95％ CI=0.46-0.90,P=0.010),and PTGS2 (HR=0.62,95％ CI=0.44-0.86,P=0.005) correlated with a higher overall survival (OS),while higher expression of MMP1 (HR=1.72,95％ CI=1.22-2.41,P=0.002),NCOA2 (HR=1.71,95％ CI=1.12-2.60,P=0.013),NOS2 (HR=1.67,95％ CI=1.20-2.32,P=0.002),and NR1I3 (HR=1.69,95％ CI=1.21-2.37,P=0.002) was associated with worse OS. The risk score was negatively correlated with the infiltration of T cells,CD8+T cells,myeloid dendritic cells,NK cells,and B cells,and positively correlated with monocyte/macrophage infiltration (all P<0.05).Conclusion:Kaempferol exerts therapeutic effects on breast cancer through multiple targets and pathways. The prognostic genes identified based on its related targets,along with the established prognostic model,can guide clinical treatment of breast cancer. The correlation between the prognostic model and immune infiltration provides direction for future experimental studies.

Objective To investigate the clinicopathological features,immunophenotypes,molecular genetic alterations,and prognosis of succinate dehydrogenase-deficient renal cell carcinoma(SDH-RCC).Methods A total of 11 cases of SDH-RCC diagnosed at West China Hospital,Sichuan University between 2016 and 2023 were selected for clinicopathological,immunohistochemical,and DNA sequencing analyses.Results Among the 11 cases of SDH-RCC,there were 5 male patients and 6 female patients.The patients'ages ranged from 12 to 71 years,with an average age of 39.7 years.Among them,5 patients had tumors located in the right kidney,5 had tumors located in the left kidney,and 1 patient had bilateral tumors.Microscopic observation showed that the tumor cells of the SDH-RCC patients displayed a wide spectrum of structures,forming sheet-like,nested,and glandular structures.In addition,tumor cells in papillary structures were observed in some cases.The tumor cells had abundant cytoplasm,was eosinophilic,and contained flocculent materials.Intracytoplasmic vacuolations were observed in some of the cells.Among all the patients,7(7/11,63.6％)showed typical low-grade features(grade 1-2 according to the International Society of Urological Pathology[ISUPJ/WHO 2016 classification),and 4(4/11,36.4％)showed high-grade features(grade 3 according to the ISUP/WHO 2016 classification).The average ages of patients with low-grade and high-grade features were 32.1 years and 58.0 years,respectively.Immunohistochemical staining of all 11 cases demonstrated negative results for SDHB and cytokeratin 7(CK7),and positive staining results for paired box 8(PAX-8),fumarate hydratase(FH),and epithelial membrane antigen(EMA).Their Ki-67 index was l％-30％.In one case,the loss of SDHB expression was also accompanied by a loss of SDHA expression.Sanger sequencing was performed to examine all the exons of SDHB in 7 cases.One case showed a frameshift mutation,c.236Tdel(p.K80Rfs*),and another case harbored a missense mutation,c.725G＞A(p.Arg242His).In another case,next generation sequencing revealed that large fragments of SDHB(Exon 4-8 del)were missing.Follow-up data were available for 10 patients.The follow-up time ranged from 4 to 138 months,with the average being 32.8 months,and all patients survived.Metastasis and recurrence were reported in 5 cases,with 3 of them showing high-grade features and 2 showing low-grade features.Conclusion SDH-RCC is rare and the patients demonstrate a relatively young age of onsets.Patients may present with bilateral tumors.Tumors with low-grade features usually occur in young patients,with their Ki-67 index usually being lower than 5％.Individual cases may experience tumor recurrence and metastasis over a long period of follow-up.Tumors with high-grade features tend to occur in older patients who have a higher Ki-67 index,and who are prone to recurrence and metastasis.Negative immunohistochemical staining results for SDHB can assist in tumor diagnosis,but the loss of SDHB protein expression does not necessarily lead to the detection of SDHB gene mutation.

Background and Aims:Kaempferol is a natural flavonoid compound that can regulate various processes and activities associated with cancer,such as the cell cycle,oxidative stress,apoptosis,proliferation,metastasis,and angiogenesis. Its potential for treating breast cancer has been validated in some studies,but its mechanism of action remains unclear. Therefore,this study was conducted to explore the potential targets and pathways of kaempferol in the treatment of breast cancer and establish a prognostic model.Methods:Using network pharmacology and bioinformatics approaches,the intersection targets of kaempferol for the treatment of breast cancer were obtained from databases such as HERB,GeneCards,STRING,PubChem,RCSB PDB,and TCGA. Protein interaction,GO analysis,and KEGG pathway enrichment analysis were performed,followed by molecular docking for validation. Prognostic-related genes were identified through LASSO-Cox regression analysis to establish a risk score model. The prognostic analysis for each gene,and the correlation between risk score and immune infiltration were analyzed. Results:Network pharmacology analysis identified 55 intersection targets. Protein interaction analysis revealed five potential key genes (Akt1,Bcl2,CASP3,ESR1,AR). GO analysis showed that kaempferol treatment of breast cancer involved 1604 biological process entries,20 cellular component entries,and 121 molecular function entries. KEGG pathway enrichment mainly included pathways related to chemical carcinogenesis,reactive oxygen species signaling,AGE-RAGE signaling in diabetic complications,TNF signaling,and IL-17 signaling. Molecular docking results indicated that kaempferol had a strong affinity for key targets,with the best binding effect observed with ESR1 (-9.1 kcal/mol). Ten prognostic genes (Bcl2,CYP1B1,DPP4,GSTM1,GSTM2,MMP1,NCOA2,NOS2,NR1I3,PTGS2) were obtained through LASSO-Cox regression analysis,and a risk score model was established. The AUC of this model for predicting breast cancer patient prognosis was greater than 0.5. Single-gene survival analysis indicated that higher expression of Bcl2 (HR=0.61,95％ CI=0.43-0.86,P=0.005),CYP1B1 (HR=0.68,95％ CI=0.49-0.94,P=0.022),GSTM1 (HR=0.68,95％ CI=0.47-0.98,P=0.037),GSTM2 (HR=0.64,95％ CI=0.46-0.90,P=0.010),and PTGS2 (HR=0.62,95％ CI=0.44-0.86,P=0.005) correlated with a higher overall survival (OS),while higher expression of MMP1 (HR=1.72,95％ CI=1.22-2.41,P=0.002),NCOA2 (HR=1.71,95％ CI=1.12-2.60,P=0.013),NOS2 (HR=1.67,95％ CI=1.20-2.32,P=0.002),and NR1I3 (HR=1.69,95％ CI=1.21-2.37,P=0.002) was associated with worse OS. The risk score was negatively correlated with the infiltration of T cells,CD8+T cells,myeloid dendritic cells,NK cells,and B cells,and positively correlated with monocyte/macrophage infiltration (all P<0.05).Conclusion:Kaempferol exerts therapeutic effects on breast cancer through multiple targets and pathways. The prognostic genes identified based on its related targets,along with the established prognostic model,can guide clinical treatment of breast cancer. The correlation between the prognostic model and immune infiltration provides direction for future experimental studies.

Transcranial magnetic stimulation (TMS) is a non-invasive neuromodulation therapy which uses short-term, rapid alternating or pulsed current in coil to produce an induced magnetic field in specific brain region. To improve its efficacy, studies on TMS targeted localization technique is advancing now. In the past 10 years, on the basis of brain-heart connection and frontal-vagal pathway theories, researchers proposed a hypothesis that neuro-cardiac-guided TMS (NCG-TMS) could be used for individualized targeting TMS treatment. A series of studies have been carried out to verify its feasibility. The aim of this review is to summarize the progress of NCG-TMS related studies.

Transcranial magnetic stimulation (TMS) is a non-invasive neuromodulation therapy which uses short-term, rapid alternating or pulsed current in coil to produce an induced magnetic field in specific brain region. To improve its efficacy, studies on TMS targeted localization technique is advancing now. In the past 10 years, on the basis of brain-heart connection and frontal-vagal pathway theories, researchers proposed a hypothesis that neuro-cardiac-guided TMS (NCG-TMS) could be used for individualized targeting TMS treatment. A series of studies have been carried out to verify its feasibility. The aim of this review is to summarize the progress of NCG-TMS related studies.

Objective:To establish a colon cancer cell line which overexpressing LINC01018 stably and study its biological characteristics.Methods:The expression of LINC01018 in HCoEpiC and HT-29 cells were detected by real time fluorescence quantitative polymerase chain reaction (qRT-PCR). HT-29 cells were infected with LINC01018 overexpression lentivirus to screen and establish HT-29 cell lines which overexpressing LINC01018 stably. The effect of LINC01018 on the proliferation, invasion and migration of HT-29 cells were detected by cell counting kit-8 (CCK-8) assay and Transwell assay separately. The expression of CDK6 and matrix metalloproteinase-2 (MMP-2) in HT-29 cells was detected by Western blot.Results:The expression of LINC01018 in HT-29 cells was significantly lower than that in the human colonic epithelial cells (HCoEpiC). HT-29-L18 cell lines which overexpressing LINC01018 stably was screened successfully. Overexpression of LINC01018 significantly inhibited the cell proliferation, invasion and migration, and reduced the protein expression of CDK6 and MMP-2 in HT-29 cells.Conclusions:The expression of LINC01018 was decreased abnormally in colon cancer cells. Up-regulation of LINC01018 expression can inhibit the proliferation, invasion and migration of colon cancer cells, which may be related to CDK6 and MMP-2.

Mantle cell lymphoma (MCL) is a distinct histological type of B-cell lymphoma with a poor prognosis. Several agents, such as proteasome inhibitors, immunomodulatory drugs, and inhibitors of B cell lymphoma-2 and Bruton's tyrosine kinase have shown efficacy for relapsed or refractory (r/r) MCL but often have short-term responses. Chimeric antigen receptor (CAR) T-cell therapy has emerged as a novel treatment modality for r/r non-Hodgkin's lymphoma. However, long-term safety and tolerability associated with CAR T-cell therapy are not defined well, especially in MCL. In this report, we described a 70-year-old patient with r/r MCL with 48-month duration of follow-up who achieved long-term remission after CAR T-cell therapy. CAR T-cell-related toxicities were also mild and tolerated well even in this elderly patient. This report suggested that CAR T-cell therapy is a promising treatment modality for patients with MCL, who are generally elderly and have comorbid conditions.
Adult ; Aged ; Cell- and Tissue-Based Therapy ; Humans ; Immunotherapy, Adoptive ; Lymphoma, Mantle-Cell/therapy* ; Neoplasm Recurrence, Local ; Receptors, Chimeric Antigen

Objective:To investigate the effects of Resolvin D1 (RvD1) on the inflammatory response and the expression of Nod-like receptor protein 3(NLRP3) inflammasomes in mice with acute lung injury.Methods:The 30 male BALB/c mice weighing 25-30 g were divided into 3 groups(each group with 10 mice). Mice in the normal control group were given normal saline by tail vein injection.Mice in the lipopolysaccharide (LPS) group were given the same volume of LPS (10 mg/kg) via tail vein injection.Mice in the RvD1 group were injected with RvD1 (5 μg/kg) through the tail vein 30 minutes prior to LPS administration.Mice were humanely sacrificed after 6 hours.Histopatholo-gical changes of lung tissue, the levels of pro-inflammatory cytokines interleukin(IL)-18 and IL-1β, and the expression of NLRP3 inflammasomes in lung tissue were measured.Results:After LPS administration, the lung of mice showed pathological damage.The levels of pro-inflammatory factors IL-18 and IL-1β as well as the expression of NLRP3, apoptosis-associated speck-like protein containing a card(ASC)and Caspase-1 in the LPS group were significantly higher than those in the normal control group (all P<0.05). After pretreatment with RvD1, the pathological damage of lung tissue was alleviated.The levels of pro-inflammatory factors IL-18 and IL-1β as well as the expression of NLRP3, ASC and Caspase-1 in the RvD1 group were significantly lower than those in the LPS group (all P<0.05). Conclusions:RvD1 can attenuate the pulmonary inflammation in acute lung injury and inhibit the release of pro-inflammatory factors, which is possibly related to the suppression of NLRP3.