Objective:To discuss the expression of tripartite motif-containing protein 24(TRIM24)in the clear cell renal cell carcinoma(ccRCC)tissue,and to clarify its relationships with the clinicopathological features and prognosis of the ccRCC patients.Methods:The cancer and paracancer normal tissues were collected from 90 ccRCC patients who had not undergone preoperative radiotherapy or chemotherapy.Tissue microarray and immunohistochemistry were used to detect the expression levels of TRIM24 protein in ccRCC tissue.The differences in TRIM24 protein expression between ccRCC and paracancer normal tissues were analyzed.The score of TRIM24 protein expression and the average value were calculated,and based on the average value,the patients were classified into TRIM24 protein low-expression and TRIM24 protein high-expression groups.The associations between the TRIM24 protein expression and different clinicopathological features of the patients were analyzed,and the relationship between the TRIM24 protein expression and the prognosis of the patients was analyzed.Results:The immunohistochemistry results showed that the TRIM24 protein was expressed in both the nucleus and cytoplasm of the ccRCC tumor cells,and there were significant differences in the TRIM24 protein expression level in ccRCC tissue when compared with paracancer normal tissue(P＜0.05).The TRIM24 protein expression in the nucleus of ccRCC tissue was associated with the patient's age,gender,and tumor size(P＜0.05).The Kaplan-Meier survival analysis results showed that the overall survivals of the patients with high TRIM24 protein expression in the cytoplasm of ccRCC tissue,older age,and higher pathological grade were shorter than those with low TRIM24 protein expression,younger age,and lower pathological grade(P＜0.05).The multivariate Cox regression analysis results showed that the prognosis of the patients with high TRIM24 protein expression in the cytoplasm and higher pathological grade were poorer compared with the patients with low TRIM24 protein expression and lower pathological grade(P＜0.05).Conclusion:The ccRCC patients with high TRIM24 expression in the cytoplasm of ccRCC tumor tissue and higher pathological grade have the lower postoperative survival rates and poorer prognosis.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Objective:To investigate the effect of asiaticoside on cognitive impairment in young epileptic rats by regulating SIRT1/AMPK signaling pathway.Methods:Young rats were randomly separated into Sham group,Model group,low-dose asiaticoside(20 mg/kg)group,high-dose asiaticoside(40 mg/kg)group and high-dose asiaticoside+EX527 group.The seizure situation of rats was observed;Morris water maze was applied to test spatial learning and memory abilities;ELISA was applied to detect levels of TNF-α,IL-10,IL-6,MDA,SOD and GSH-Px in rat hippocampal tissue;Nissl staining was applied to observe morphological changes of rat hippocampal tissue;TUNEL staining was applied to detect apoptosis of hippocampal neurons in rats;Western blot was applied to detect expressions of Cleaved-caspase-3,Bax,Bcl-2,SIRT1,p-AMPK and AMPK proteins in rat hippocampal tissue.Results:Compared with Sham group,Model group showed severe tissue damage in hippocampal CA1 region of rats,epilepsy score and seizure duration were obviously increased,incubation period was obviously prolonged,levels of TNF-α,IL-6,MDA,neuronal apoptosis rate,expressions of Cleaved-caspase-3 and Bax proteins in CA1 region of hippocampus were obviously increased,the retention time and number of platform crossings were obviously decreased,levels of IL-10,SOD,GSH-Px,protein expressions of Bcl-2,SIRT1,p-AMPK/AMPK in CA1 region of hippocampus were obviously reduced(P<0.05);compared with Model group,tissue damage in hippocampal CA1 region of rats in low-dose and high-dose asiaticoside groups was obviously reduced,the epilepsy score and seizure duration were obviously decreased,incubation period was obviously shortened,levels of TNF-α,IL-6,MDA,neuronal apoptosis rate,expressions of Cleaved-caspase-3 and Bax proteins in CA1 region of hippocampus were obviously reduced,the retention time and the number of platform crossings were obviously increased,levels of IL-10,SOD,GSH-Px,protein expressions of Bcl-2,SIRT1 and p-AMPK/AMPK in CA1 region of hippocampus were obviously increased(P<0.05);EX527 was able to partially reverse the improve-ment effect of asiaticoside on epileptic young rats.Conclusion:Asiaticoside can reduce inflammatory response,oxidative stress and hippocampal neuronal apoptosis in rats and reduce nerve damage by activating SIRT1/AMPK signaling pathway,thus improving cogni-tive function of young epileptic.

Objective:To investigate the effect of asiaticoside on cognitive impairment in young epileptic rats by regulating SIRT1/AMPK signaling pathway.Methods:Young rats were randomly separated into Sham group,Model group,low-dose asiaticoside(20 mg/kg)group,high-dose asiaticoside(40 mg/kg)group and high-dose asiaticoside+EX527 group.The seizure situation of rats was observed;Morris water maze was applied to test spatial learning and memory abilities;ELISA was applied to detect levels of TNF-α,IL-10,IL-6,MDA,SOD and GSH-Px in rat hippocampal tissue;Nissl staining was applied to observe morphological changes of rat hippocampal tissue;TUNEL staining was applied to detect apoptosis of hippocampal neurons in rats;Western blot was applied to detect expressions of Cleaved-caspase-3,Bax,Bcl-2,SIRT1,p-AMPK and AMPK proteins in rat hippocampal tissue.Results:Compared with Sham group,Model group showed severe tissue damage in hippocampal CA1 region of rats,epilepsy score and seizure duration were obviously increased,incubation period was obviously prolonged,levels of TNF-α,IL-6,MDA,neuronal apoptosis rate,expressions of Cleaved-caspase-3 and Bax proteins in CA1 region of hippocampus were obviously increased,the retention time and number of platform crossings were obviously decreased,levels of IL-10,SOD,GSH-Px,protein expressions of Bcl-2,SIRT1,p-AMPK/AMPK in CA1 region of hippocampus were obviously reduced(P<0.05);compared with Model group,tissue damage in hippocampal CA1 region of rats in low-dose and high-dose asiaticoside groups was obviously reduced,the epilepsy score and seizure duration were obviously decreased,incubation period was obviously shortened,levels of TNF-α,IL-6,MDA,neuronal apoptosis rate,expressions of Cleaved-caspase-3 and Bax proteins in CA1 region of hippocampus were obviously reduced,the retention time and the number of platform crossings were obviously increased,levels of IL-10,SOD,GSH-Px,protein expressions of Bcl-2,SIRT1 and p-AMPK/AMPK in CA1 region of hippocampus were obviously increased(P<0.05);EX527 was able to partially reverse the improve-ment effect of asiaticoside on epileptic young rats.Conclusion:Asiaticoside can reduce inflammatory response,oxidative stress and hippocampal neuronal apoptosis in rats and reduce nerve damage by activating SIRT1/AMPK signaling pathway,thus improving cogni-tive function of young epileptic.

OBJECTIVE To study the effects of methimazole on the urinary metabolomics of hyperthyroidism rats， and to preliminarily investigate its possible mechanism. METHODS Thirty SD rats were randomly divided into control group， model group and methimazole group， with 10 rats in each group. Except for the control group， the rats in the other two groups were given Levothyroxine sodium tablets 160 mg/kg by intragastric administration for 15 days； at the same time， methimazole group was additionally given methimazole 3.6 mg/kg daily by intragastric administration every day. The basic condition of the rats was observed， and the body weight and anal temperature were measured. After the last medication， the serum levels of triiodothyronine （T3）， tetraiodothyronine （T4）， free triiodothyronine （FT3）， free tetraiodothyronine （FT4）， and thyroid stimulating hormone （TSH） were determined； 24-hour urine was collected on the 15th day after administration. UPLC-TOF-MS was used to analyze the urine metabolomics of rats. Principal component analysis and orthogonal partial least squares-discriminant analysis were used to screen out related differential metabolites， and potential metabolic pathways were analyzed by using HMDB and KEGG. RESULTS Compared with the control group， the rectal temperature， serum levels of T3， T4， FT3 and FT4， the expressions of differential metabolites sebacic acid， cholic acid 3-O-glucuronic acid and N6， N6， N6-trimethyl-L-lysine in urine were significantly up-regulated， while body weight， serum level of TSH， the expressions of deoxycytidine and 2-oxo-4-methylthiobutanoic acid in urine were significantly down-regulated （P＜0.01）. Compared with model group， above indexes of rats were reversed significantly in methimazole group （P＜0.01 or P＜0.05）. Above five differential metabolites were mainly involved in four signaling pathways: pentose and glucuronate interaction， lysine degradation， cysteine and methionine metabolism， and pyrimidine metabolism. CONCLUSIONS Methimazole might improve hyperthyroidism by modulating the four pathways of pentose and glucuronate interaction， lysine degradation， cysteine and methionine metabolism， and pyrimidine metabolism.

Objective To compare seminal carnitine levels between normal males and asthenozoospermic patients,evaluate its correla-tion with progressive motility(PR)of sperm,and observe the effects of exogenous carnitine supplementation on asthenozoospermic pa-tients.Methods Semen samples were collected from 511 normal fertile males and asthenozoospermic patients.Seminal was measured using a fixed-time assay kit and the levels of carnitine were compared between the two groups.The consistency between seminal carni-tine and PR was assessed.Additionally,77 asthenozoospermic patients received L-carnitine(1 g/time,3 times/day,30 days/course).The levels of seminal carnitine and PR alteration pre-and post-treatment were monitored.Results The seminal L-carnitine level in the patients with asthenospermia([194.34±65.41]μmol/L)was significantly lower than that in normal fertile males([405.43±72.12]μmol/L)(P＜0.01).When the seminal L-carnitine level ≥325 μmol/L was set as the threshold,the statistical results showed that Kappa value was 0.81,with a diagnostic coincidence rate of 93.74％.After one course of administration of L-carnitine,the concentra-tion of seminal L-carnitine([356.03±84.87]μmol/L)and PR([32.69±8.35]％)were significantly higher those that before treat-ment([183.61±79.54]μmol/L and[16.56±7.74]％,P＜0.01).Conclusion The seminal carnitine assay kit could be used for ac-curate and high-throughput quantification of clinical samples,facilitating asthenozoospermia diagnosis and therapeutic efficacy evalua-tion.Exogenous carnitine supplementation may elevate seminal carnitine levels and sperm motility in asthenozoospermic patients and po-tentially improve their fertility.

Malignant tumors are diseases that seriously threaten human health and social development. Traditional tumor therapies such as surgery, radiotherapy, chemotherapy and targeted therapy cannot fully meet the needs of clinical treatment, and emerging immunotherapy has become a research hotspot in the field of tumor treatment. Immune checkpoint inhibitors (ICIs) have been approved as a tumor immunotherapy method for the treatment of various tumors, such as lung cancer, liver cancer, stomach cancer and colorectal cancer, etc. However, during the clinical use of ICIs, only a small number of patients experienced durable responses, which also led to drug resistance and adverse reactions. Therefore, the identification and development of predictive biomarkers is crucial to improve the therapeutic efficacy of ICIs. The predictive biomarkers of tumor ICIs mainly include tumor biomarkers, tumor microenvironment biomarkers, circulation-related biomarkers, host environmental biomarkers and combinatorial biomarkers. They are of great significance for screening, individualized treatment and prognosis evaluation of tumor patients. This article reviews the advances of predictive markers for tumor ICIs therapy.
Humans ; Immune Checkpoint Inhibitors/therapeutic use* ; Lung Neoplasms ; Biomarkers ; Immunotherapy/methods* ; Biomarkers, Tumor/genetics* ; Prognosis ; Tumor Microenvironment

The estimated new cases of breast cancer (BC) patients were 2.26 million in 2020, which accounted for 11.7% of all cancer patients, making it the most prevalent cancer worldwide. Early detection, diagnosis and treatment are crucial to reduce the mortality, and improve the prognosis of BC patients. Despite the widespread use of mammography screening as a tool for BC screening, the false positive, radiation, and overdiagnosis are still pressing issues that need to be addressed. Therefore, it is urgent to develop accessible, stable, and reliable biomarkers for non-invasive screening and diagnosis of BC. Recent studies indicated that the circulating tumor cell DNA (ctDNA), carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA15-3), extracellular vesicles (EV), circulating miRNAs and BRCA gene from blood, and the phospholipid, miRNAs, hypnone and hexadecane from urine, nipple aspirate fluid (NAF) and volatile organic compounds (VOCs) in exhaled gas were closely related to the early screening and diagnosis of BC. This review summarizes the advances of the above biomarkers in the early screening and diagnosis of BC.
Humans ; Female ; Biomarkers, Tumor ; Early Detection of Cancer ; Breast Neoplasms/diagnosis* ; Prognosis ; MicroRNAs/genetics*

Tumor is a serious threat to human health. At present, surgical resection, chemoradiotherapy, targeted therapy and immunotherapy are the main therapeutic strategies. Monoclonal antibody has gradually become an indispensable drug type in the clinical treatment of cancer due to its high efficiency and low toxicity. Phage antibody library technology (PALT) is a novel monoclonal antibody preparation technique. The recombinant immunoglobulin variable region of heavy chain (VH)/variable region of light chain (VL) gene is integrated into the phage vector, and the antibody is expressed on the phage surface in the form of fusion protein to obtain a diverse antibody library. Through the process of adsorption-elution-amplification, the antibody library can be screened to obtain the antibody molecule with specific binding antigen as well as its gene sequence. PALT has the advantages of short antibody production cycle, strong plasticity of antibody structure, large antibody yield, high diversity and direct production of humanized antibodies. It has been used in screening tumor markers and preparation of antibody drugs for breast cancer, gastric cancer, lung cancer and liver cancer. This article reviews the recent progress and the application of PALT in tumor therapy.
Humans ; Bacteriophages/genetics* ; Immunoglobulin Variable Region/genetics* ; Gene Library ; Antibodies, Monoclonal/therapeutic use* ; Immunotherapy ; Peptide Library

Objective　To investigate the correlation between red cell distribution width to platelet ratio and short-term prognosis in acute ischemic stroke.Methods　The patients with ischemic stroke admitted in the First Affiliated Hospital of Zhengzhou University during the initial 24 hours after the onset of disease between 2015-2017 were recruited prospectively.Red cell distribution width (RDW),platelet and clinical characteristics were recorded.Functional outcome at 90 days after ischemic stroke was evaluated by the modified Rankin Scale (mRS).Results　Totally 1106 patients were included.Patients with poor functional outcome were older (P＜0.001=and had more frequent histories of hypertension (P=0.015),diabetes (P=0.001),atrial fibrillation (P＜0.001=,coronary heart disease (P=0.041),stroke (P＜0.001=and higher National Institutes of Health Stroke Scale (NIHSS)(P＜0.001=,white blood cell count (P＜0.001=,RDW (P＜0.001=and RDW to platelet ratio (RPR) (P=0.034).Higher RPR levels were associated with poor functional outcome at 90 days after ischemic stroke［(odds ratio,OR) 1.09,(confidence interval,CI) 1.03~1.16，P=0.005］. Conclusion　Higher RPR levels were associated with poor short-term outcome after acute ischemic stroke.