Objective To investigate the effect of abnormal iron metabolism on neurological func-tion in elderly patients with hypertensive intracerebral hemorrhage(HICH)after minimally inva-sive surgical evacuation.Methods A prospective study was conducted on 300 elderly patients with HICH admitted to our hospital from January 2021 to December 2023.At 28 d after surgery,Glas-gow Outcome Scale(GOS)was used to assess the presence of neurological deficits or not,and then they were divided into a good neurological function group(GOS score≥4,175 cases)and a poor neurological function group(GOS score＜4,125 cases).Iron deposition in cerebrospinal fluid and serum iron metabolism were compared between the two groups,and the risk factors for neu-rological deterioration were analyzed.Results Compared with the good neurological function group,the poor neurological function group exhibited significantly decreases in Glasgow coma scale(GCS)scores at admission and 7 d after admission and iron ions(P＜0.01).Bleeding vo-lume,ferritin,transferrin,and quantitative susceptibility mapping(QSM)values of the thalamus and the hippocampus were obviously increased upon admission(P＜0.01).Multi variate logistic regression analysis showed that admission bleeding volume(OR=1.083,95％CI:1.012-1.159,P=0.021),ferritin(OR=1.065,95％CI:1.016-1.116,P=0.009),and thalamic QSM value(OR=4.075,95％CI:2.848-5.830,P=0.000)were risk factors for neurological dysfunction in the HICH patients after minimally invasive surgical treatment,while GCS score(OR=0.430,95％CI:0.259-0.715,P=0.001)and iron ions(OR=0.193,95％CI:0.064-0.581,P=0.003)at 7 d of admission were protective factors.Conclusion Iron deposition in cerebrospinal fluid and abnormal iron metabolism in serum are related to the deterioration of neurological function in eld-erly patients with HICH after minimally invasive hematoma evacuation,and are regarded as po-tential therapeutic targets.

Objective To investigate the effect of abnormal iron metabolism on neurological func-tion in elderly patients with hypertensive intracerebral hemorrhage(HICH)after minimally inva-sive surgical evacuation.Methods A prospective study was conducted on 300 elderly patients with HICH admitted to our hospital from January 2021 to December 2023.At 28 d after surgery,Glas-gow Outcome Scale(GOS)was used to assess the presence of neurological deficits or not,and then they were divided into a good neurological function group(GOS score≥4,175 cases)and a poor neurological function group(GOS score＜4,125 cases).Iron deposition in cerebrospinal fluid and serum iron metabolism were compared between the two groups,and the risk factors for neu-rological deterioration were analyzed.Results Compared with the good neurological function group,the poor neurological function group exhibited significantly decreases in Glasgow coma scale(GCS)scores at admission and 7 d after admission and iron ions(P＜0.01).Bleeding vo-lume,ferritin,transferrin,and quantitative susceptibility mapping(QSM)values of the thalamus and the hippocampus were obviously increased upon admission(P＜0.01).Multi variate logistic regression analysis showed that admission bleeding volume(OR=1.083,95％CI:1.012-1.159,P=0.021),ferritin(OR=1.065,95％CI:1.016-1.116,P=0.009),and thalamic QSM value(OR=4.075,95％CI:2.848-5.830,P=0.000)were risk factors for neurological dysfunction in the HICH patients after minimally invasive surgical treatment,while GCS score(OR=0.430,95％CI:0.259-0.715,P=0.001)and iron ions(OR=0.193,95％CI:0.064-0.581,P=0.003)at 7 d of admission were protective factors.Conclusion Iron deposition in cerebrospinal fluid and abnormal iron metabolism in serum are related to the deterioration of neurological function in eld-erly patients with HICH after minimally invasive hematoma evacuation,and are regarded as po-tential therapeutic targets.

Objective To construct and validate an effective prognostic nomogram for the patients with incidental gallbladder cancer(IGBC).Methods The clinical data of 161 patients with IGBC requiring radical surgery admitted to the First Affiliated Hospital of Xi'an Jiaotong University from May 2011 to October 2022 was analyzed retrospectively.COX proportional risk regression model was used to screen for influencing factors on overall survival(OS)of IGBC.Nomogram was constructed based on independent influencing factors that affected the prognosis of IGBC patients.The concordance index(C-index)and calibration curve were used to validate the performance of the model.Receiver operating characteristic(ROC)curve analysis and decision curve analysis(DCA)were used to validate the predictive accuracy and net benefit of the plotted column chart.Results Univariate COX regression analysis suggested that age,T stage,N stage,M stage,preoperative carcinoembryonic antigen(CEA),preoperative carbohydrate antigenl9-9(CA19-9),preoperative red blood cell volume distribution on width coefficient of variation(RDW-CV),treatment method,and recurrence and metastasis were risk factors which affected the long-term survival of IGBC patients after radical surgery.Multivariate COX regression analysis suggested that T stage,N stage,preoperative CA19-9,preoperative RDW-CV,preoperative AST,treatment methods,and recurrence and metastasis were independent risk factors which affected the prognosis of IGBC patients.The C-index of the constructed prognostic model was 0.872.The calibration plot demonstrated good performance of the Nomogram.ROC curve analysis showed an area under the curve of 0.869,confirming a high sensitivity and specificity.A high net benefit was proven by DCA.Conclusions The constructed Nomogram.can accurately and intuitively predict the survival probability of IGBC patients after radical surgery.

Pancreatic cancer has a very poor prognosis.Early diagnosis and treatment are especially critical for improving its prognosis.Nanotechnology has been widely used in the diagnosis and treatment of pancreatic cancer.Relying on the unique physicochemical properties of nanoparticles and their rich surface modifications,effective enrichment of tumor sites can be achieved.Magnetic iron oxide nanoparticles(MIONPs)is one of the commonly used nanomaterials in the diagnosis and treatment of pancreatic cancer,and has good biocompatibility.Through special surface modification,it can be used in targeted diagnosis and treatment of pancreatic cancer.MIONPs can be used as a contrast agent for MRI,and by modifying the surface,they also can be used in targeted imaging of pancreatic cancer.And they can also be modified as a drug delivery system to achieve targeted delivery of drugs and improve therapeutic effects.However,the application of MIONPs in pancreatic cancer diagnosis and treatment still faces some challenges,such as nanotoxicity and cost issues.With the development of technology,MIONPs are expected to play an important role in the personalized diagnosis and treatment of pancreatic cancer.

Background and purpose:Pancreatic cancer is a highly aggressive solid tumor of the digestive system,with radical resection being unfeasible in approximately 80%of patients due to the absence of specific clinical manifestations in the early stages.The use of gemcitabine as a first-line chemotherapeutic agent has not significantly improved patient prognosis,primarily due to the development of chemoresistance.The precise mechanisms underlying gemcitabine resistance in pancreatic cancer remain unclear.This study aimed to explore potential biomarkers associated with gemcitabine chemoresistance in pancreatic cancer by utilizing gemcitabine-resistant cell lines and pathological pancreatic cancer tissues,in conjunction with data from online databases.Additionally,we analyzed follow-up data from pancreatic cancer patients to assess the impact of relevant targets on patient prognosis.Methods:In this study,gemcitabine-resistant cell lines were developed through stepwise induction using a gemcitabine concentration gradient.Second-generation high-throughput RNA-seq sequencing was conducted on these resistant cells,and bioinformatics analysis was employed to identify four pancreatic cancer genes from the Gene Expression Omnibus(GEO)datasets(GSE106336,GSE110580,GSE35141,and GSE140077).Co-expressed genes were screened using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot and immunohistochemistry to verify the expression levels of target molecules.Surgical specimens from 70 patients diagnosed with pancreatic adenocarcinoma at the First Affiliated Hospital of Xi'an Jiao Tong University between June 2018 and June 2021 were analyzed.These included 30 specimens resistant to gemcitabine,16 non-resistant specimens,and 24 normal pancreatic tissues as controls.Ethical approval was obtained(Ethical approval:2021 LunxuanKeZi No.457,No.XJTU1AF2021LSK-457).Clinical and prognostic information was collected,and the log-rank test was used to evaluate the relationship between target molecule expression and patient prognosis.Results:The half maximal inhibitory concentration(IC50)for gemcitabine was significantly higher in the gemcitabine-resistant cell strain(Mia GR)than in the wild-type cell line(Mia WT)(258.10 μmol/L vs 0.18 μmol/L),with a resistance index(RI)of 1 443.9.Transcriptome sequencing identified 3 985 differentially expressed genes,of which 25 were shared with the GEO datasets.Further analysis highlighted INPP4B as a key gene.RTFQ-PCR and Western blot confirmed that INPP4B mRNA and protein levels were significantly elevated in drug resistant cells compared to wild-type cells(P＜0.05).Immunohistochemical analysis revealed that INPP4B expression was significantly higher in drug resistant pancreatic cancer tissues compared to non-drug resistant tissues,and lower in normal tissues than in both cancerous tissue types.Kaplan-Meier curves demonstrated that patients with low INPP4B expression had significantly better progression-free survival(PFS)than those with high expression(HR=2.874,95%CI:1.262-6.544,P=0.013).Although patients with low INPP4B expression also showed better overall survival(OS),the difference was not statistically significant(HR=1.484,95%CI:0.518-4.250,P=0.465).Conclusion:INPP4B may serve as a potential biomarker for gemcitabine chemoresistance in pancreatic cancer and is associated with poor prognosis in drug resistant patients.Developing targeted assays and treatments for INPP4B could facilitate early identification of patients likely to exhibit resistance to gemcitabine therapy,thereby improving their prognosis.

Objective:To investigate the effect of copper chlorophyllin sodium salt(CHL)on the sensitivity of human pancreatic cancer cells in response to gemcitabine(GEM)therapy and on the therapeutic effect on pancreatic cancer cells that have developed GEM resistance.Methods:MIA GR(a pancreatic cancer cell line resistant to GEM)was induced by a low-dose continuous incremental method,and the half inhibitory concentration(IC50)of MIA WT and MIA GR to GEM treatment was detected by the CCK-8 method,and the resistance index was calculated;the difference in IC50 of CHL on the two types of cells was detected by the CCK-8 method after treating MIA WT and MIA GR cells with different concentrations of CHL,CCK-8 method was used to detect the difference in IC50 of CHL on the two types of cells;on the basis of IC50,MIA WT and MIA GR cells were intervened with CHL and(or)GEM with different multiplicity of IC50,respectively,and the growth inhibition curves of MIA WT and MIA GR cells were detected by the CCK-8 method under the intervention of CHL combined with GEM;After the intervention of MIA WT and MIA GR cells with CHL and(or)GEM at IC50,respectively,the effects on the proliferation of the two different cells were detected using the clone formation assay;the effects on cytotoxicity/activity were observed under fluorescence microscopy;and the effects on apoptosis were detected using flow cytometry.Finally,western blotting was used to detect the effects of CHL and(or)GEM interventions on the drug resistance-associated molecules P-glycoprotein(P-gp)and ribonucleotide reductase regulatory subunit M2(RRM2)in MIA GR cells,the and sensitivity-related molecule deoxycytidine kinase(DCK)on protein expression levels.Results:MIA GR cells were verified to be well drug resistant,with resistance indices of 549.1 and 667.9 after 48 h and 96 h after GEM intervention compared to homologous wild-type MIA WT cells,respectively;CHL intervention inhibited the proliferation of MIA GR cells more significantly compared to that of MIA WT cells;and CHL in combination with GEM exerted a more significant growth inhibitory effect compared to GEM alone in both MIA WT cells(P<0.001)and MIA GR cells(P<0.01).CHL significantly inhibited the tumor proliferation of MIA GR cells,and the inhibitory effect was more pronounced in both cells when combined with GEM(P<0.000 1);furthermore,compared to GEM alone,the intervention with CHL could cause more pronounced cytotoxicity(P<0.000 1)in both MIA WT and MIA GR cells.caused more pronounced cytotoxicity(P<0.000 1)and induced a higher percentage of apoptosis than GEM alone.The results of the western blotting assay showed that CHL intervention caused a decrease in the expression levels of P-gp and RRM2 proteins,as well as an increase in the protein expression level of DCK in MIA GR cells.Conclusion:CHL increases the sensitivity of pancreatic cancer cells to GEM and also induces a decrease in the resistance of drug-resistant pancreatic cancer cells to GEM.

The purpose of this study was to reveal the resistance mechanism of Mycoplasma bovis from Xizang yaks to macrolide antibiotics and provide theoretical basis for clinical medication.In this study,10 strains of Mycoplasma bovis from Xizang yaks were tested for drug sensitivity to macrolide antibiotics using the micromethod,and sensitive strains were induced to be highly resist-ant in vitro.The results showed that all 10 strains of Mycoplasma bovis exhibited varying degrees of resistance to macrolide antibiotics.However,through methylation enzyme and inactivation en-zyme gene detection and analysis,it was found that there was no methylation enzyme encoded by the methylation enzyme gene and no resistance mechanism mediated by macrolide inactivation en-zymes,indicating that there were no mutations in the target gene loci of the sensitive and resistant strains.However,highly resistant strains in vitro have mutations at the domain Ⅱ,L22,and even L4 target gene loci,indicating that if two or more target gene amino acid loci undergo mutations,highly resistant strains can be produced.After testing with active efflux systems,it was found that there were no active efflux systems using macrolide antibiotics as substrates.It can be seen that the strain of Mycoplasma bovis from Xizang yaks is prone to mutation under the pressure of high con-centrations of macrolides,and there are mutations in two or more target gene amino acid sites,which is prone to produce highly resistant strains.

Background and purpose:Pancreatic cancer is a highly aggressive solid tumor of the digestive system,with radical resection being unfeasible in approximately 80%of patients due to the absence of specific clinical manifestations in the early stages.The use of gemcitabine as a first-line chemotherapeutic agent has not significantly improved patient prognosis,primarily due to the development of chemoresistance.The precise mechanisms underlying gemcitabine resistance in pancreatic cancer remain unclear.This study aimed to explore potential biomarkers associated with gemcitabine chemoresistance in pancreatic cancer by utilizing gemcitabine-resistant cell lines and pathological pancreatic cancer tissues,in conjunction with data from online databases.Additionally,we analyzed follow-up data from pancreatic cancer patients to assess the impact of relevant targets on patient prognosis.Methods:In this study,gemcitabine-resistant cell lines were developed through stepwise induction using a gemcitabine concentration gradient.Second-generation high-throughput RNA-seq sequencing was conducted on these resistant cells,and bioinformatics analysis was employed to identify four pancreatic cancer genes from the Gene Expression Omnibus(GEO)datasets(GSE106336,GSE110580,GSE35141,and GSE140077).Co-expressed genes were screened using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot and immunohistochemistry to verify the expression levels of target molecules.Surgical specimens from 70 patients diagnosed with pancreatic adenocarcinoma at the First Affiliated Hospital of Xi'an Jiao Tong University between June 2018 and June 2021 were analyzed.These included 30 specimens resistant to gemcitabine,16 non-resistant specimens,and 24 normal pancreatic tissues as controls.Ethical approval was obtained(Ethical approval:2021 LunxuanKeZi No.457,No.XJTU1AF2021LSK-457).Clinical and prognostic information was collected,and the log-rank test was used to evaluate the relationship between target molecule expression and patient prognosis.Results:The half maximal inhibitory concentration(IC50)for gemcitabine was significantly higher in the gemcitabine-resistant cell strain(Mia GR)than in the wild-type cell line(Mia WT)(258.10 μmol/L vs 0.18 μmol/L),with a resistance index(RI)of 1 443.9.Transcriptome sequencing identified 3 985 differentially expressed genes,of which 25 were shared with the GEO datasets.Further analysis highlighted INPP4B as a key gene.RTFQ-PCR and Western blot confirmed that INPP4B mRNA and protein levels were significantly elevated in drug resistant cells compared to wild-type cells(P＜0.05).Immunohistochemical analysis revealed that INPP4B expression was significantly higher in drug resistant pancreatic cancer tissues compared to non-drug resistant tissues,and lower in normal tissues than in both cancerous tissue types.Kaplan-Meier curves demonstrated that patients with low INPP4B expression had significantly better progression-free survival(PFS)than those with high expression(HR=2.874,95%CI:1.262-6.544,P=0.013).Although patients with low INPP4B expression also showed better overall survival(OS),the difference was not statistically significant(HR=1.484,95%CI:0.518-4.250,P=0.465).Conclusion:INPP4B may serve as a potential biomarker for gemcitabine chemoresistance in pancreatic cancer and is associated with poor prognosis in drug resistant patients.Developing targeted assays and treatments for INPP4B could facilitate early identification of patients likely to exhibit resistance to gemcitabine therapy,thereby improving their prognosis.

Objective:To investigate the effect of copper chlorophyllin sodium salt(CHL)on the sensitivity of human pancreatic cancer cells in response to gemcitabine(GEM)therapy and on the therapeutic effect on pancreatic cancer cells that have developed GEM resistance.Methods:MIA GR(a pancreatic cancer cell line resistant to GEM)was induced by a low-dose continuous incremental method,and the half inhibitory concentration(IC50)of MIA WT and MIA GR to GEM treatment was detected by the CCK-8 method,and the resistance index was calculated;the difference in IC50 of CHL on the two types of cells was detected by the CCK-8 method after treating MIA WT and MIA GR cells with different concentrations of CHL,CCK-8 method was used to detect the difference in IC50 of CHL on the two types of cells;on the basis of IC50,MIA WT and MIA GR cells were intervened with CHL and(or)GEM with different multiplicity of IC50,respectively,and the growth inhibition curves of MIA WT and MIA GR cells were detected by the CCK-8 method under the intervention of CHL combined with GEM;After the intervention of MIA WT and MIA GR cells with CHL and(or)GEM at IC50,respectively,the effects on the proliferation of the two different cells were detected using the clone formation assay;the effects on cytotoxicity/activity were observed under fluorescence microscopy;and the effects on apoptosis were detected using flow cytometry.Finally,western blotting was used to detect the effects of CHL and(or)GEM interventions on the drug resistance-associated molecules P-glycoprotein(P-gp)and ribonucleotide reductase regulatory subunit M2(RRM2)in MIA GR cells,the and sensitivity-related molecule deoxycytidine kinase(DCK)on protein expression levels.Results:MIA GR cells were verified to be well drug resistant,with resistance indices of 549.1 and 667.9 after 48 h and 96 h after GEM intervention compared to homologous wild-type MIA WT cells,respectively;CHL intervention inhibited the proliferation of MIA GR cells more significantly compared to that of MIA WT cells;and CHL in combination with GEM exerted a more significant growth inhibitory effect compared to GEM alone in both MIA WT cells(P<0.001)and MIA GR cells(P<0.01).CHL significantly inhibited the tumor proliferation of MIA GR cells,and the inhibitory effect was more pronounced in both cells when combined with GEM(P<0.000 1);furthermore,compared to GEM alone,the intervention with CHL could cause more pronounced cytotoxicity(P<0.000 1)in both MIA WT and MIA GR cells.caused more pronounced cytotoxicity(P<0.000 1)and induced a higher percentage of apoptosis than GEM alone.The results of the western blotting assay showed that CHL intervention caused a decrease in the expression levels of P-gp and RRM2 proteins,as well as an increase in the protein expression level of DCK in MIA GR cells.Conclusion:CHL increases the sensitivity of pancreatic cancer cells to GEM and also induces a decrease in the resistance of drug-resistant pancreatic cancer cells to GEM.

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.