Objective:To investigate the affection of high mobility group nucleosome-binding protein 5(HMGN5) gene in the hepatocellular cancer tissue and HepG2 cell and its function in the growth of HepG2 cell.Methods:70 Patients with hepatocellular carcinoma who under-went surgical resection and were confirmed by postoperative pathology in Sunshine Union Hospital of Weifang City from January 2017 to June 2020 were selected. Their clinical and pathological data, surgical resection of liver cancer tissue and adjacent normal liver tissue were collected, their survival time were also recorded. The contents of HMGN5 protein in the hepatocellular cancer tissues and adjacent normal liver tissues of 70 patients were detected by immunohistochemistry. Compare the expression of HMGN5 protein in liver cancer tissue and normal liver tissue and the positive expression rate of HMGN5 protein in liver cancer tissues of hepatocellular carcinoma patients with different clinical characteristics, to analyze the relationship between HMGN5 expression and prognosis of patients with hepatocellular carcinoma.The mRNA contents of HMGN5 gene in HepG2 cells and HL-7702 cells were determined by quantitative real time polymerase chain reaction(qRT-PCR) method. After transfectHepG2 cells with HMGN5 shRNA, the proliferation ability of HepG2 cells were evaluated by cell proliferation assay and the cell apoptosis was analyzed by flow cytometry. Measurement data with normal distribution were expressed as ±s, two independent samples t test was used for comparison between groups. Counting data was expressed as n(%), χ2 test was used for comparison between groups. Survival analysis of patients was performed by Kaplan-Meier method. Results:Immunohistochemical tests showed that HMGN5 staining is strong in liver cancer tissue, but weak in normal liver tissue. HMGN5 protein expression was positive in 48 of 70 patients with hepatocellular carcinoma. The positive expression rate of HMGN5 protein in hepatocellular carcinoma tissues of patients with pathological grade 3+4 and clinical stage Ⅲ+Ⅳ was higher than that of patients with grade 1+2 and stage Ⅰ+Ⅱ, respectively. There was no significant difference in cumulative survival rate between HMGN5 positive expression group and HMGN5 negative expression group ( χ2=3.81, P=0.051). The results of qRT-PCR showed that the expression level of HMGN5 mRNA in hepatoma HepG2 cells was higher than that in normal liver HL-7702 cells [(4.51±0.45) vs (1.35±0.27), the difference was statistically significant ( t=10.43, P=<0.001). After 24, 48 and 72h of the HMGN5 gene was knockout, the survival rate of HepG2 hepatoma cells was lower than that of HL-7702 hepatoma cells (all P<0.05) and the apoptosis rate of hepatoma HepG2 cells was higher than that of liver HL-7702 cells [(18.63±1.76)%vs(1.81±0.21)%] ,the difference was statistically significant ( t=16.44, P<0.001). Conclusions:HMGN5 genes and proteins are highly expressed in the hepatocellular cancer tissues and HepG2 cells. HMGN5 gene plays an important role in the growth of hepatocellular cancer and can be used as a potential target of treatment for hepatocellular cancer.

Objective To identify determinants of subtherapeutic voriconazole(VRCZ)concentrations in allogeneic hematopoietic stem cell transplantation(allo-HSCT)recipients and to develop/validate a nomogram-based risk prediction model.Methods This study retrospectively analyzed 310 VRCZ therapeutic drug monitoring(TDM)measurements from allo-HSCT recipients at 310 patients who under went allo-HSCT surgery at Hebei Yanda Ludaopei Hospital from October 2022 to October 2024 and received VRCZ for the prevention and treatment of invasive fungal infections before transplantion were selected as the study subjects.Cases were stratified into target-concentration group(0.5～5.0μg/ml)and subtherapeutic group(<0.5μg/ml).Through single factor and multiple factor Logistic regression analysis,indeipendent predictive factors forvecz plasma concentration non-compliance were screened,and a column chart prediction model(NPM)was constructed.The performance of the model was evaluateding area under the receiver operating characteristic curve(AUC),Hosmer-Lemeshow(H-L)goodness-of-fit test,and decision curve analysis(DCA).Results Among 310 VRCZ-TDM measurements,71.61%(222/310)achieved target concentrations.Multivariate analysis showed that CYP2C19 intermediate metabolite,daily dose of cyclosporine A(CSA),daily dose of VRCZ,creatinine(Cr)>97 μmol/L,albumin(Alb)and C-reactive protein(CRP)were independent influencing factors for VRCZ blood drug concentration non-compliance(Wald χ2=4.046～13.221,all P＜0.05).The nomogram demonstrated excellent discrimination,calibration(H-L goodness of fit test χ2=2.663,P=0.954),and clinical utility with net benefit across 0.05～0.96 risk thresholds.Conclusion The nomogram incorporating CYP2C19 gene phenotype,daily CSA dosing,daily VRCZ dosing,Cr levels,Alb and CRP provides a validated tool for optimizing VRCZ therapy in allo-HSCT recipients,enabling precision dosing strategies.

Objective To identify determinants of subtherapeutic voriconazole(VRCZ)concentrations in allogeneic hematopoietic stem cell transplantation(allo-HSCT)recipients and to develop/validate a nomogram-based risk prediction model.Methods This study retrospectively analyzed 310 VRCZ therapeutic drug monitoring(TDM)measurements from allo-HSCT recipients at 310 patients who under went allo-HSCT surgery at Hebei Yanda Ludaopei Hospital from October 2022 to October 2024 and received VRCZ for the prevention and treatment of invasive fungal infections before transplantion were selected as the study subjects.Cases were stratified into target-concentration group(0.5～5.0μg/ml)and subtherapeutic group(<0.5μg/ml).Through single factor and multiple factor Logistic regression analysis,indeipendent predictive factors forvecz plasma concentration non-compliance were screened,and a column chart prediction model(NPM)was constructed.The performance of the model was evaluateding area under the receiver operating characteristic curve(AUC),Hosmer-Lemeshow(H-L)goodness-of-fit test,and decision curve analysis(DCA).Results Among 310 VRCZ-TDM measurements,71.61%(222/310)achieved target concentrations.Multivariate analysis showed that CYP2C19 intermediate metabolite,daily dose of cyclosporine A(CSA),daily dose of VRCZ,creatinine(Cr)>97 μmol/L,albumin(Alb)and C-reactive protein(CRP)were independent influencing factors for VRCZ blood drug concentration non-compliance(Wald χ2=4.046～13.221,all P＜0.05).The nomogram demonstrated excellent discrimination,calibration(H-L goodness of fit test χ2=2.663,P=0.954),and clinical utility with net benefit across 0.05～0.96 risk thresholds.Conclusion The nomogram incorporating CYP2C19 gene phenotype,daily CSA dosing,daily VRCZ dosing,Cr levels,Alb and CRP provides a validated tool for optimizing VRCZ therapy in allo-HSCT recipients,enabling precision dosing strategies.

Objective:To investigate the affection of high mobility group nucleosome-binding protein 5(HMGN5) gene in the hepatocellular cancer tissue and HepG2 cell and its function in the growth of HepG2 cell.Methods:70 Patients with hepatocellular carcinoma who under-went surgical resection and were confirmed by postoperative pathology in Sunshine Union Hospital of Weifang City from January 2017 to June 2020 were selected. Their clinical and pathological data, surgical resection of liver cancer tissue and adjacent normal liver tissue were collected, their survival time were also recorded. The contents of HMGN5 protein in the hepatocellular cancer tissues and adjacent normal liver tissues of 70 patients were detected by immunohistochemistry. Compare the expression of HMGN5 protein in liver cancer tissue and normal liver tissue and the positive expression rate of HMGN5 protein in liver cancer tissues of hepatocellular carcinoma patients with different clinical characteristics, to analyze the relationship between HMGN5 expression and prognosis of patients with hepatocellular carcinoma.The mRNA contents of HMGN5 gene in HepG2 cells and HL-7702 cells were determined by quantitative real time polymerase chain reaction(qRT-PCR) method. After transfectHepG2 cells with HMGN5 shRNA, the proliferation ability of HepG2 cells were evaluated by cell proliferation assay and the cell apoptosis was analyzed by flow cytometry. Measurement data with normal distribution were expressed as ±s, two independent samples t test was used for comparison between groups. Counting data was expressed as n(%), χ2 test was used for comparison between groups. Survival analysis of patients was performed by Kaplan-Meier method. Results:Immunohistochemical tests showed that HMGN5 staining is strong in liver cancer tissue, but weak in normal liver tissue. HMGN5 protein expression was positive in 48 of 70 patients with hepatocellular carcinoma. The positive expression rate of HMGN5 protein in hepatocellular carcinoma tissues of patients with pathological grade 3+4 and clinical stage Ⅲ+Ⅳ was higher than that of patients with grade 1+2 and stage Ⅰ+Ⅱ, respectively. There was no significant difference in cumulative survival rate between HMGN5 positive expression group and HMGN5 negative expression group ( χ2=3.81, P=0.051). The results of qRT-PCR showed that the expression level of HMGN5 mRNA in hepatoma HepG2 cells was higher than that in normal liver HL-7702 cells [(4.51±0.45) vs (1.35±0.27), the difference was statistically significant ( t=10.43, P=<0.001). After 24, 48 and 72h of the HMGN5 gene was knockout, the survival rate of HepG2 hepatoma cells was lower than that of HL-7702 hepatoma cells (all P<0.05) and the apoptosis rate of hepatoma HepG2 cells was higher than that of liver HL-7702 cells [(18.63±1.76)%vs(1.81±0.21)%] ,the difference was statistically significant ( t=16.44, P<0.001). Conclusions:HMGN5 genes and proteins are highly expressed in the hepatocellular cancer tissues and HepG2 cells. HMGN5 gene plays an important role in the growth of hepatocellular cancer and can be used as a potential target of treatment for hepatocellular cancer.

Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

Drug-associated reward memories are conducive to intense craving and often trigger relapse. Simvastatin has been shown to regulate lipids that are involved in memory formation but its influence on other cognitive processes is elusive. Here, we used a mass spectrometry-based lipidomic method to evaluate the impact of simvastatin on the mouse brain in a cocaine-induced reinstatement paradigm. We found that simvastatin blocked the reinstatement of cocaine-induced conditioned place preference (CPP) without affecting CPP acquisition. Specifically, only simvastatin administered during extinction prevented cocaine-primed reinstatement. Global lipidome analysis showed that the nucleus accumbens was the region with the greatest degree of change caused by simvastatin. The metabolism of fatty-acids, phospholipids, and triacylglycerol was profoundly affected. Simvastatin reversed most of the effects on phospholipids induced by cocaine. The correlation matrix showed that cocaine and simvastatin significantly reshaped the lipid metabolic pathways in specific brain regions. Furthermore, simvastatin almost reversed all changes in the fatty acyl profile and unsaturation caused by cocaine. In summary, pre-extinction treatment with simvastatin facilitates cocaine extinction and prevents cocaine relapse with brain lipidome remodeling.
Animals ; Brain ; Cocaine ; Conditioning, Operant ; Extinction, Psychological ; Lipidomics ; Male ; Mice ; Simvastatin/therapeutic use*

Drug-associated reward memories are conducive to intense craving and often trigger relapse. Simvastatin has been shown to regulate lipids that are involved in memory formation but its influence on other cognitive processes is elusive. Here, we used a mass spectrometry-based lipidomic method to evaluate the impact of simvastatin on the mouse brain in a cocaine-induced reinstatement paradigm. We found that simvastatin blocked the reinstatement of cocaine-induced conditioned place preference (CPP) without affecting CPP acquisition. Specifically, only simvastatin administered during extinction prevented cocaine-primed reinstatement. Global lipidome analysis showed that the nucleus accumbens was the region with the greatest degree of change caused by simvastatin. The metabolism of fatty-acids, phospholipids, and triacylglycerol was profoundly affected. Simvastatin reversed most of the effects on phospholipids induced by cocaine. The correlation matrix showed that cocaine and simvastatin significantly reshaped the lipid metabolic pathways in specific brain regions. Furthermore, simvastatin almost reversed all changes in the fatty acyl profile and unsaturation caused by cocaine. In summary, pre-extinction treatment with simvastatin facilitates cocaine extinction and prevents cocaine relapse with brain lipidome remodeling.

Objective To explore the clinical value of CEUS guided biopsy of lymphoma in anterior mediastinum.Methods The data of 36 patients with lymphoma of anterior mediastinum underwent biopsy guided by CEUS and 36 patients by conventional ultrasound retrospectively.The successful rate of biopsy and rate of complication occurence were compared between the CEUS group and conventional ultrasound group.Results The successful rate of biopsy in CEUS group was 100％ (36/36),including 26 non-Hodgkin's lymphoma (NHL),10 Hodgkin's lymphoma (HL).The total times of puncture were 60 in 36 patients.The rate of complication occurrence was 11.11 ％ (4/36).The successful rate in conventional ultrasound group was 88.89％ (32/36),including 22 NHL,14 HL.The times of puncture were 91 in 36 patients.The rate of complication occurrence was 41.67％ (15/36).There were significant differences in successful rate and the rate of complication occurrence between two groups (x2 =4.235,8.651,P=0.040,0.003).Conclusion CEUS can reflect the microcirculation of lymphomas in anterior mediastinum,and can guide targeted biopsy.It can improve the successful rate of biopsy and reduce the complications.

Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.