Objective To explore the correlation of MET status in patients with advanced prostatic acinar adenocarcinoma with the clinical pathological parameters and prognosis. Methods The specimen from 135 patients with advanced prostatic acinar adenocarcinoma was included. The expression of c-MET protein was detected via immunohistochemistry, and MET gene amplification was assessed by fluorescence in situ hybridization. The relationships of c-MET expression and gene amplification with clinicopathological features and prognosis were analyzed. Results The positive expression rate of c-MET was 52.60% (71/135). Compared with the c-MET expression in adjacent tissues, that in tumor tissues showed lower heterogeneous expression. Among the cases, 1.71% (2/117) exhibited MET gene polyploidy, but no gene amplification was detected. Positive c-MET expression was significantly correlated with high Gleason scores and grade groups (P=0.0073). However, no significant relationship was found between c-MET expression and progression-free survival or post-treatment t-PSA levels. Conclusion c-MET protein is expressed at a relatively low level in advanced prostatic acinar adenocarcinoma, suggesting that c-MET may not be a viable target for ADC drug development in prostatic acinar adenocarcinoma.

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction

Objective To analyze the relationship of total imaging burden score with blood pressure variability(BPV)and cognitive function in elderly patients with cerebral small vessel disease(CSVD).Methods Clinical data of 182 elderly CSVD patients admitted in our hospital from December 2022 to January 2024 were collected and analyzed retrospectively.According to the results of Chinese Guidelines for Diagnosis and Treatment of CSVD-Related Cognitive Dysfunc-tion(2019),Montreal Cognitive Assessment(MoCA)and other tools for their cognitive impair-ment,they were divided into cognitively impaired group(76 cases)and cognitively normal group(106 cases).The total burden score and BPV indicators[24 h systolic blood pressure coefficient of variation(24 h SBPCV),24 h diastolic blood pressure coefficient of variation(24 h DBPCV)]were compared between the two groups.ROC curve was plotted to evaluate the diagnostic value of total burden score,24 h SBPCV and 24 h DBPCV on cognitive impairment in elderly CSVD patients.Multivariate logistic regression analysis was used to identify the risk factors of cognitive impairment in the patients.Based on the MoCA score of the cognitively impaired group,these pa-tients were further assigned into mild and moderate-to-severe cognitive impairment subgroups[with a MoCA score of 18-25(43 cases)and＜18(33 cases),respectively].Then the total burden score,24 h SBPCV and 24 h DBPCV were compared between the two subgroups.Pearson correla-tion coefficient was adopted to evaluate the correlation of severity of cognitive impairment with total burden score,24 h SBPCV and 24 h DBPCV in the elderly CSVD patients.Results The total burden score,24 h SBPCV and 24 h DBPCV were significantly higher in the cognitively impaired group than the cognitively normal group(P＜0.01).ROC curve analysis indicated that total bur-den score(AUC=0.953,95％CI:0.926-0.980,P=0.000),24 h SBPCV(AUC=0.850,95％CI:0.795-0.906,P=0.000)and 24 h DBPCV(AUC=0.761,95％CI:0.690-0.832,P=0.000)had good diagnostic efficiency for cognitive impairment in the elderly CSVD patients,with a cut-off value of 1.5,11.82％,and 8.92％,respectively.Multivariate logistic regression analysis revealed that the above three indicators were risk factors for cognitive impairment in the elderly patients with CSVD(P＜0.05,P＜0.01).Their values were significantly lower in the mild than the moder-ate-to-severe cognitive impairment subgroups(P＜0.01).Pearson correlation analysis displayed that MoCA score was negatively correlated with total burden score,24 h SBPCV and 24 h DBPCV in elderly patients with CSVD(r=-0.755,-0.632,-0.601,P＜0.01).Conclusion Detection of total burden score and BPV indicators is beneficial to the assessment of cognitive impairment in elderly CSVD patients.The higher the total burden score and the greater the BPV indicators,the more severe the cognitive impairment is,which may explore new ideas for clinical diagnosis and treatment of CSVD.

Objective Melatonin has been shown to have neuroprotective effects.This study is aimed at observing the effects of copper deposition on cognitive function in a toxic milk(TX)mouse model of Wilson disease(WD),and investigating the effects and mechanisms of action of Gandou Bushen Decoction(GDBSD)on melatonin synthesis and pineal function in the WD model mice.Methods A total of 30 homozygous TX mice were randomly assigned to 3 groups(n=10 in each group),including a WD group,a GDBSD group,and a dimercaptosuccinic acid(DMSA)group.A total of 10 DL mice were included in the normal control(NC)group.The structure and copper content of pineal gland tissues,oxidative stress and apoptosis-related markers,and serum melatonin levels were evaluated using hematoxylin-eosin(HE)staining,enzyme-linked immunosorbent assay(ELISA),flow cytometry,and Western blot.Results Compared with the NC group,the WD group exhibited decreased learning and cognitive abilities(P＜0.05),damaged pineal gland structure,increased copper content,reactive oxygen species(ROS)levels,and mitochondrial damage rate in the pineal gland(P＜0.01),altered levels of melatonin and oxidative stress-related markers(P＜0.05),upregulated expression levels of pro-apoptotic proteins Bax and Caspase-3,and decreased expression of the anti-apoptotic protein Bcl-2(P＜0.01).After treatment with GDBSD and DMSA,the SIRT3/FOXO3α signaling pathway was activated,the copper content in the pineal gland was reduced,and oxidative stress and apoptosis-related damages were improved,leading to an improvement in learning and memory abilities(P＜0.05).Conclusion GDBSD can alleviate cognitive impairments in WD mice caused by pineal gland copper deposition by inhibiting oxidative stress and apoptosis in the pineal gland.The underlying molecular mechanism is associated with the regulation of the SIRT3/FOXO3α signaling pathway.

Objective:To investigate the predictive value of neutrophil extracellular traps (NETs) on the prognosis of patients with epidermal growth factor receptor (EGFR) wild-type lung adenocarcinoma.Methods:A total of 132 surgical paraffin specimens of EGFR wild-type lung adenocarcinoma diagnosed at the Tumor Hospital Affiliated to Xinjiang Medical University from January 2016 to December 2023 and 12 pairs of cancer and paracancerous fresh tissues from patients with EGFR wild-type lung adenocarcinoma diagnosed in March-July 2024 were collected with their clinical information. Western blotting and immunofluorescence were used to detect the expression levels of citrullinated histone H3 (CitH3) and myeloperoxidase (MPO) in cancerous and paraneoplastic tissues. Immunohistochemistry was used to detect the infiltration of PD-L1, CD4 + T cells and CD8 + T cells in the tumors. The clinical and prognostic correlations between NETs and EGFR wild-type lung adenocarcinoma patients were analyzed. Results:The expression of MPO ( P＜0.001) and CitH3 ( P=0.009) was significantly increased in the tumors compared with the paracancerous tissues. The rate of high expression of NETs in cancer tissues was higher in patients with EGFR wild-type lung adenocarcinoma who were in stage Ⅲ and Ⅳ, with lymph node metastasis, distant metastasis, pleural invasion, high expression of Ki-67, low expression of CD8 + T, and lowered lymphocyte counts when compared to paraneoplastic tissues ( P＜0.05). Patients were stratified based on TNM stage Ⅱb for prognostic analysis. Kaplan-Meier univariate analysis showed that the median overall survival (OS) (stage Ⅰ to Ⅱb: 47 vs 87 months; stage Ⅲ to Ⅳ: 27 months vs not reach) and the median disease-free survival (DFS) (stage Ⅰ to Ⅱb: 42 vs 78 months; stage Ⅲ to Ⅳ: 18 vs 39 months) of patients with high expression of NETs in stage Ⅰ to II b and stage Ⅲ to Ⅳ were lower than those with low expression (stage Ⅰ to Ⅱb: OS, P＜0.001; DFS, P＜0.001; stage Ⅲ to Ⅳ: OS, P=0.001; DFS, P=0.022). The OS (stage Ⅰ to Ⅱb: HR=3.513, 95% CI: 1.966-6.277, P＜0.001; stage Ⅲ to Ⅳ: HR=3.215, 95% CI: 1.324-7.806, P=0.010) and the DFS (stage Ⅰ to Ⅱb: HR=2.478 ,95% CI: 1.396-4.400, P=0.002; stage Ⅲ to Ⅳ: HR=2.248, 95% CI: 1.089-4.638, P=0.028) in the group with high expression of NETs in either stage Ⅰ to Ⅱb or stage Ⅲ to Ⅳ were significantly shorter than those in the group with low expression. Conclusion:The EGFR wild-type lung adenocarcinoma patients with high expression of NETs have relatively shorter DFS and OS, which are independent risk factors for the prognosis of patients with EGFR wild-type lung adenocarcinoma, and are likely to be the potential biomarkers for the diagnosis and treatment of EGFR wild-type lung adenocarcinoma.

Objective:To investigate affinity of monoclonal antibody(MAb)of different cardiac troponinⅠ(cTnⅠ)epi-topes to cTnⅠ-troponin C(cTnⅠ-TnC)antigen,and to screen novel cTnⅠ-specific antibodies for clinical detection kit development.Methods:Based on technology platform of microarray chemiluminescence immunoassay,cTnⅠ-TnC antigen spot sample hard matrix chip was used,mouse derived cTnⅠ MAb was used as antibody to be detected,HRP-sheep-anti-mouse-IgG was used as antibody to detect,and experimental conditions were optimized for anti-origin solution,antigen spot sample concentration and antibody to be detected concentration.Affinity of cTnⅠ MAb with different antigenic epitopes to cTnⅠ-TnC was detected by competition method.Graded concentrations of cTnⅠ-TnC were added to experimental group,and diluents of equal volume were added to control group.Reaction signals of different cTnⅠ MAb were detected and affinity constants were calculated.Results:Antigen spot sample liquid was P105,antigen spot sample concentration was 0.1 mg/ml,and antibody concentration to be detected was 80 ng/ml were optimal experi-mental conditions.20c6cc and 7B9cc MAb containing TnC antibodies had the best affinity,and 20c6cc MAb reached 3.18×106 L/mol.cTnⅠ MAb located at C-end of peptide chain in a.a.r 130～145,169～178 and 190～196 regions showed good affinity,among which MAb 625(a.a.r 169～178)was 6.37×105 L/mol,showing the strongest affinity among cTnⅠ MAb.Conclusion:MAb has good affinity with cTnⅠ-TnC antigen at C-end of cTnⅠ peptide chain a.a.r 130～145,169～178,190～196 and TnC region.Designing a new type of cTnⅠ complement antibody targeting this region can be a new way to solve negative interference of autoantibody and peptide proteolysis.

Objective::This study aimed to assess the impact of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor treatment immediately after percutaneous coronary intervention (PCI) on the myocardial salvage index (MSI) in patients with anterior ST-segment elevation myocardial infarction (STEMI) 5-10 d after the procedure.Methods::The early PCSK9 inhibitor thERapy Following pErcutaneous Coronary inTervention (PERFECT) trial is a prospective randomized controlled trial. From January 2021 to December 2023, 32 patients with anterior STEMI from Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and Shanghai Tenth People’s Hospital were enrolled in the PERFECT trial. Patients were randomly assigned in a 1∶1 ratio to the PCSK9 inhibitor group ( n = 16) or the control group ( n = 16), and their baseline data were collected. Patients in the PCSK9 inhibitor group (ie, alirocumab group) received a subcutaneous injection of PCSK9 inhibitor (alirocumab, 75 mg) immediately after PCI based on conventional treatment. In the control group, patients received only conventional treatment. The primary endpoint was the MSI measured by cardiovascular magnetic resonance 5-10 d after PCI. The secondary endpoints included the left ventricular ejection fraction measured by cardiovascular magnetic resonance 5-10 d after PCI and the time to peak of creatine kinase isoenzyme-MB and high-sensitivity cardiac troponin T. Safety endpoints included any clinical adverse events that occurred during the 6-month follow-up period. Results::Baseline data during admission showed no intergroup significance. No significant difference in MSI (55.54% ± 14.80% vs. 44.72% ± 15.42%, P = 0.056) and left ventricular ejection fraction (51.24% ± 8.91% vs. 44.99% ± 8.84%, P = 0.060) was observed. Additional, there was no significant difference in the time to peak of creatine kinase isoenzyme-MB ((12.97 ± 5.67) h vs. (14.31 ± 7.04) h, P = 0.557) and high-sensitivity cardiac troponin T ((21.03 ± 12.46) h vs. (21.44 ± 9.99) h, P = 0.920) between the 2 groups. During the 6-month follow-up period, only 1 patient in the PCSK9 inhibitor group developed cerebral hemorrhage 6 months after PCI. Conclusions::Early treatment with alirocumab did not exhibit a significant increase in MSI at 5-10 d in patients with anterior STEMI. Larger trials are necessary to evaluate the impact of early administration of PCSK9 inhibitors after myocardial infarction.

Staphylococcus aureus has the ability to invade bovine mammary epithelial cells and cause persistent intracellular infections.For diagnosing mastitis in cows caused by the bacterial in-fections,it is necessary to determine whether there is a persistent intracellular infection by S.au-reus within mammary epithelial cells,and to establish scientific treatment strategies and imple-ment proactive management decisions.The content of this study is the establishment of a brand new diagnostic method aimed at confirming this type of bovine disease efficiently and cost-effec-tively.The diagnostic procedure is as follows:Add 100 μL S.aureus liquid culture with a concentra-tion of 8.5 ×1010 CFU/mL to the well-grown bovine mammary epithelial cell line and co-culture at 37 degrees Celsius for 2 h to establish an intracellular infection model;After the co-cultivation is completed,add 100 μL of lysostaphin to the dish to thoroughly eliminate extracellular S.aureus.It can be confirmed that the model has been successfully established that black spherical bacteria with a diameter of approximately 1μm are observed within the sample cells using a transmission e-lectron microscope.Intracellular infection is evidenced by the observation of DNA-staining positive material within the cytoplasm of DAPI-stained sample cells,as observed through a laser confocal microscope.The sample is subjected to gram staining after being treated with 200 μL of 0.5％Tri-tonX-100 for 15 min,and intracellular infection is observed within the cytoplasm of the sample cells with Gram-positive results under oil immersion microscopy.The results of the novel"Tri-tonX-100 & gram staining method"validate that the presence of Staphylococcus aureus individu-als within bovine mammary epithelial cells is consistent with images observed under transmission electron microscopy and laser confocal microscopy.The verification results indicate that the"Tri-tonX-100 & gram staining method"is suitable for the diagnosis of S.aureus persistent intracellu-lar infection in bovine mastitis.Later,somatic cells from milk were extracted by gradient centrifu-gation and purified to obtain bovine mammary epithelial cells for clinical testing of 70 S.aureus positive mastitis cattle,which showed a positive rate of 82.9％and a negative rate of 17.1％.

Purpose To explore the clinical value of real-time three-dimensional automatic left atrial quantification technology in evaluating left atrial volume and function in heart failure with preserved ejection fraction(HFpEF).Materials and Methods A total of 65 patients diagnosed as HFpEF at the Fourth Affiliated Hospital of Harbin Medical University from December 2021 to October 2023 were prospectively enrolled.The control group included 65 healthy subjects who underwent ultrasound examination during the same period and were matched with the HFpEF group in terms of age and gender.According to the New York Heart Association(NYHA)cardiac function classification,patients with NYHA grade Ⅰ+Ⅱ were classified into the HFpEF group A,and those with grade Ⅲ+Ⅳ into the HFpEF group B.Relevant clinical data,conventional ultrasound parameters and three-dimensional ultrasound parameters were recorded in both the HFpEF group and the control group.Left atrial volume parameters,longitudinal strain parameters and circumferential strain parameters were analyzed.The area under the receiver operating characteristic curve was used to compare the diagnostic efficacy of left atrial functional parameters for HFpEF.Results Compared with the control group,the HFpEF group exhibited significant abnormalities in cardiac structure and function.Specifically,left ventricular posterior wall thickness,interventricular septal thickness at end-diastole,and mean E/e′ were significantly increased(t=-5.127,-5.886,-16.670,all P<0.05),while the absolute value of left ventricular global longitudinal strain(LVGLS)and septal and lateral mitral annular e′ were significantly decreased(t=-17.092,40.279,45.412,all P<0.05).All left atrial volume parameters were significantly increased,whereas left atrial functional and strain parameters were significantly decreased(t=-13.632-6.912,all P<0.05).Compared with HFpEF group A,HFpEF group B showed lower left atrial total emptying fraction,left atrial expansion index,left atrial contraction strain and absolute value of LVGLS(t=2.062,3.545,-2.189,-2.586),as well as a higher left atrial minimum volume(t=-2.187),respectively(all P<0.05).Left atrial reservoir strain was negatively correlated with N-terminal pro-B-type natriuretic peptide and mean E/e′(r=-0.395,-0.626,both P<0.05),and positively correlated with the absolute value of LVGLS(r=0.602,P<0.05).The LASr and LAEI had high predictive value for HFpEF,with area under the curve of 0.898 and 0.817,cut-off values of 20.5%and 112%,sensitivities of 96.9%and 83.1%,specificities of 75.4%and 78.5%,and Youden indices of 0.723 and 0.616,respectively.Conclusion Real-time three-dimensional automatic left atrial quantification technology enables early and sensitive detection of left atrial dysfunction in HFpEF.Among the parameters derived,LASr(a strain parameter)and LAEI(a functional parameter)exhibit high diagnostic efficacy for HFpEF.

Objective::This study aimed to assess the impact of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor treatment immediately after percutaneous coronary intervention (PCI) on the myocardial salvage index (MSI) in patients with anterior ST-segment elevation myocardial infarction (STEMI) 5-10 d after the procedure.Methods::The early PCSK9 inhibitor thERapy Following pErcutaneous Coronary inTervention (PERFECT) trial is a prospective randomized controlled trial. From January 2021 to December 2023, 32 patients with anterior STEMI from Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and Shanghai Tenth People’s Hospital were enrolled in the PERFECT trial. Patients were randomly assigned in a 1∶1 ratio to the PCSK9 inhibitor group ( n = 16) or the control group ( n = 16), and their baseline data were collected. Patients in the PCSK9 inhibitor group (ie, alirocumab group) received a subcutaneous injection of PCSK9 inhibitor (alirocumab, 75 mg) immediately after PCI based on conventional treatment. In the control group, patients received only conventional treatment. The primary endpoint was the MSI measured by cardiovascular magnetic resonance 5-10 d after PCI. The secondary endpoints included the left ventricular ejection fraction measured by cardiovascular magnetic resonance 5-10 d after PCI and the time to peak of creatine kinase isoenzyme-MB and high-sensitivity cardiac troponin T. Safety endpoints included any clinical adverse events that occurred during the 6-month follow-up period. Results::Baseline data during admission showed no intergroup significance. No significant difference in MSI (55.54% ± 14.80% vs. 44.72% ± 15.42%, P = 0.056) and left ventricular ejection fraction (51.24% ± 8.91% vs. 44.99% ± 8.84%, P = 0.060) was observed. Additional, there was no significant difference in the time to peak of creatine kinase isoenzyme-MB ((12.97 ± 5.67) h vs. (14.31 ± 7.04) h, P = 0.557) and high-sensitivity cardiac troponin T ((21.03 ± 12.46) h vs. (21.44 ± 9.99) h, P = 0.920) between the 2 groups. During the 6-month follow-up period, only 1 patient in the PCSK9 inhibitor group developed cerebral hemorrhage 6 months after PCI. Conclusions::Early treatment with alirocumab did not exhibit a significant increase in MSI at 5-10 d in patients with anterior STEMI. Larger trials are necessary to evaluate the impact of early administration of PCSK9 inhibitors after myocardial infarction.