OBJECTIVE To investigate the protective effect and mechanism of saikosaponin C （SSC） on acute liver injury （ALI） in mice induced by carbon tetrachloride （CCl4） based on serum metabolomics. METHODS Forty mice were divided into blank group （water）， model group （water）， positive control drug group （Biphenyl diester drop pills， 150 mg/kg）， and SSC low- and high-dose groups （2.5， 10 mg/kg） using the random number table method， with 8 mice in each group. They were given water/ relevant drugs， once a day， for 7 consecutive days. One hour after the last administration， all mice were intraperitoneally injected with 0.2% CCl4 olive oil to induce ALI model， except for the blank group. After 17 hours of the modeling， the liver index of mice was calculated. The levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， lactate dehydrogenase （LDH）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and IL-1β in serum of mice were detected. The histopathological changes of liver tissue were observed. Meanwhile， the serum metabolomics of mice were analyzed by liquid chromatography-mass spectrometry. RESULTS Compared with the blank group， the levels of liver index， ALT， AST， LDH， TNF-α， IL-6， and IL-1β in the model group were significantly increased （P＜0.01）. Hepatocytes were edema， vacuolar degeneration， more necrosis， and a large number of inflammatory cells were infiltrated. Compared with the model group， liver index and serum index levels of mice were significantly decreased （P＜0.05 or P＜0.01）， accompanied by marked improvement in histopathological damage to the liver tissue. The metabolomics results showed that compared with the model group， there were 63 up-regulated and 256 down-regulated differential metabolites in the serum of mice in the SSC high-dose group， including prostaglandin B2， 20-hydroxy-leukotriene B4， 5- hydroxy-L-tryptophan， 7α -hydroxycholesterol， etc.； these metabolites were primarily involved in metabolic pathways such as arachidonic acid metabolism， 5-hydroxytryptamine synapse， primary bile acid biosynthesis. CONCLUSIONS SSC exerts a protective effect against CCl4-induced ALI by down-regulating the level of key metabolites such as prostaglandin B2 and 20-hydroxy-leukotriene B4， and then ruducing metabolic pathways such as arachidonic acid metabolism， 5- hydroxytryptamine synapse， and primary bile acid biosynthesis.

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

Cancer multidrug resistance (MDR) impairs the therapeutic efficacy of various chemotherapeutics. Novel approaches, particularly the development of MDR reversal agents, are critically needed to address this challenge. This study demonstrates that tenacissoside I (TI), a compound isolated from Marsdenia tenacissima (Roxb.) Wight et Arn, traditionally used in clinical practice as an ethnic medicine for cancer treatment, exhibits significant MDR reversal effects in ABCB1-mediated MDR cancer cells. TI reversed the resistance of SW620/AD300 and KBV200 cells to doxorubicin (DOX) and paclitaxel (PAC) by downregulating ABCB1 expression and reducing ABCB1 drug transport function. Mechanistically, protein arginine methyltransferase 1 (PRMT1), whose expression correlates with poor prognosis and shows positive association with both ABCB1 and EGFR expressions in tumor tissues, was differentially expressed in TI-treated SW620/AD300 cells. SW620/AD300 and KBV200 cells exhibited elevated levels of EGFR asymmetric dimethylarginine (aDMA) and enhanced PRMT1-EGFR interaction compared to their parental cells. Moreover, TI-induced PRMT1 downregulation impaired PRMT1-mediated aDMA of EGFR, PRMT1-EGFR interaction, and EGFR downstream signaling in SW620/AD300 and KBV200 cells. These effects were significantly reversed by PRMT1 overexpression. Additionally, TI demonstrated resistance reversal to PAC in xenograft models without detectable toxicities. This study establishes TI's MDR reversal effect in ABCB1-mediated MDR human cancer cells through inhibition of PRMT1-mediated aDMA of EGFR, suggesting TI's potential as an MDR modulator for improving chemotherapy outcomes.
Humans ; Protein-Arginine N-Methyltransferases/antagonists & inhibitors* ; Drug Resistance, Neoplasm/drug effects* ; ErbB Receptors/genetics* ; Animals ; Cell Line, Tumor ; Drug Resistance, Multiple/drug effects* ; Methylation/drug effects* ; Saponins/administration & dosage* ; Mice ; Mice, Nude ; Mice, Inbred BALB C ; ATP Binding Cassette Transporter, Subfamily B/genetics* ; Doxorubicin/pharmacology* ; Paclitaxel/pharmacology* ; Female ; Repressor Proteins

Objective:To discuss the effect of sericin on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-κB(NF-κB)signaling pathway and apoptosis in the streptozotocin(STZ)-damaged INS-1 cells,and to clarify its mechanism.Methods:The INS-1 cells were cultured with complete medium containing 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 Akt1 inhibitor A-674563,10 mmol·L-1 STZ,and 600 mg·L-1 sericin,and divided into 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563 groups,and the control group(complete medium without drugs)was set up.Cell counting kit-8(CCK-8)method was used to detect the survival rates of the INS-1 cells,and the half-maximal inhibitory concentration(IC50)value was calculated to determine the optimal inhibitory concentration of A-674563,which was further verified by Western blotting method.The INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),and low,medium,and high doses of sericin groups(10 mmol·L-1 STZ+150 mg·L-1 sericin+complete medium,10 mmol·L-1 STZ+300 mg·L-1 sericin+complete medium,and 10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium).CCK-8 method was used to detect the survival rates of the INS-1 cells in various groups to determine the optimal concentration of sericin.Additionally,the INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),sericin group(10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium),and A-674563 group(10 mmol·L-1 STZ+600 mg·L-1 sericin+0.3 μmol·L-1 A-674563+complete medium).Flow cytometry was used to detect the apoptotic rates of the INS-1 cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Akt1,NF-κB,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the INS-1 cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated Akt1(p-Akt1)and NF-κB proteins in the INS-1 cells in various groups;enzyme linked immunosorbent assay(ELISA)method was used to detect the levels of TNF-α and IL-6 in the INS-1 cells in various groups.Results:The survival rates of the INS-1 cells in control group was 100.00%±0.00%;in 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium groups,which were 82.50%±2.28%,69.47%±1.94%,51.51%±1.74%,38.94%±1.57%,24.79%±1.14%,and 19.85%±1.03%,respectively.The IC?? value of A-674563 for INS-1 cells was 0.3 μmol·L-1,and 0.3 μmol·L-1 A-674563 was selected for subsequent experiments.Compared with 0 μmol·L-1 A-674563,the expression level of p-Akt1 protein in the INS-1 cells after treated with 0.3 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium was significantly decreased(P<0.05).The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rates of the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the survival rate of the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Thus,600 mg·L-1 sericin was selected for subsequent experiments.The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rate of the INS-1 cells in sericin group was significantly increased(P<0.05);compared with sericin group,the survival rate of the INS-1 cells in A-674563 group was significantly decreased(P<0.05).The flow cytometry results showed that compared with normal control group,the apoptotic rate of the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the apoptotic rate of the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the apoptotic rate of the INS-1 cells in A-674563 group was significantly increased(P<0.05).The RT-qPCR results showed that compared with normal control group,the expression level of Akt1 mRNA in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of Akt1 mRNA in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of Akt1 mRNA in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB mRNA in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The Western blotting results showed that compared with normal control group,the expression level of p-Akt1 protein in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of p-Akt1 protein in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of p-Akt1 protein in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression level of NF-κB protein in the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the expression level of NF-κB protein in the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB protein in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The ELISA results showed that compared with normal control group,the levels of TNF-α and IL-6 in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α and IL-6 in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the levels of TNF-α and IL-6 in the INS-1 cells in A-674563 group were significantly increased(P<0.05).Conclusion:Sericin alleviates the PI3K/Akt/NF-κB signaling pathway-mediated inflammatory response and apoptosis by targeting Akt1,exerting a protective effect against STZ-induced damage in INS-1 cells.

The elimination of biofilms is a crucial step in controlling hospital-acquired infections.Once biofilms colonize luminal instruments,it is difficult to remove them using traditional disinfection methods.Conventional disinfection approaches now face a series of challenges,including microbial resistance,corrosiveness,cytotoxicity,residual disinfection byproducts,and environmental pollution.Therefore,developing novel disinfection technologies specifically targeting biofilm removal is vitally important.New disinfection technologies,such as slightly acidic electrolyzed water,plasma technology,surface modification techniques,nanomaterial-based disinfection,bacteriophage disinfection,and enzymatic disinfection,are constantly emerging.These technologies exhibit excellent performance against biofilms by leveraging the synergistic effects of multiple mechanisms,including the reactive oxygen species(ROS)burst,photocatalytic oxidation,physical disruption,and biological targeting.This review summarizes the characteristics,underlying mechanisms,and potential application scenarios of these novel disinfection technologies,with a particular focus on their effects against biofilms formed by common pathogenic bacteria on surfaces in hospital settings.It aims to provide a reference basis for the practical application and translation of these disinfection technologies and the development of new disinfection strategies.

Objective To analyze the development of nuclear medicine services in Hunan Province and to assess internal radiation doses among the nuclear medicine workers (NMWs) involved in iodine-131 radionuclide therapy. Methods Based on a survey of nuclear medicine institutions in Hunan Province, a total of 61 NMWs from seven hospitals providing iodine-131 therapy for thyroid cancer were selected as the study subjects by convenience sampling method. Thyroidal iodine-131 activity was measured using a portable gamma spectrometer to estimate internal dose and total annual effective dose. Results A total of 47 nuclear medicine institutions were reported in Hunan Province by 2023, most of which were public and tertiary hospitals, accounting for 38. Iodine-131 therapy was performed in 30 institutions, including nine for thyroid cancer. A total of nine participants had detectable thyroidal iodine-131 activity among 61 workers involved in iodine-131 thyroid cancer treatment, with the detection rate of 14.8%. Their internal radiation annual committed effective doses ranged from 0.100 to 1.584 mSv, with a mean of 0.499 mSv and median of 0.426 mSv. Except for one cleaner, the remaining eight physicians and nurses had the total annual effective doses ranging from 0.311 to 3.007 mSv, with a mean of 1.305 mSv, all below the annual dose limit of 20.000 mSv among radiation workers specified in national standard. Conclusion Internal exposure to iodine-131 among the NMWs should not be neglected. Standardized procedures and strengthened internal dose monitoring are recommended.

Frailty is a complex age-related clinical condition characterized by a decline in physiological capacity across several organ systems, with a resultant increased susceptibility to stressors.The relationship between frailty assessment and elderly patients’ safety during perioperative period has received widespread attention.This paper reviews research progress of frailty assessment in elderly patients during perioperative period in the department of urology.

OBJECTIVE To investigate the damage effect and potential toxic mechanism of SARS-CoV-2 spike protein(S protein)on human neuroblastomacells(SH-SY5Y).METHODS SH-SY5Y were treated with S protein at concentrations of 25,50,75,and 100 mg·L-1 for 24 h.Cell viability of SH-SY5Y was detected using the CCK-8 assay.The cytotoxic lactate dehydrogenase(LDH)detection kit was used to measure the release rate of LDH,and the 5-ethynyl-2′-deoxyuridine(EdU)-488 cell prolifera-tion kit was used to assess cell proliferation.The ATP detection kit was used to measure intracellular ATP content.The JC-1 fluorescent probe method was employed to detect the mitochondrial membrane potential(MMP)of cells.Seahorse XF was used to measure mitochondrial respiratory and glycolytic capacity.RESULTS Compared with the cell control group,cell viability was significantly reduced in S protein 25,50,75 and 100 mg·L-1 groups(P<0.01),and the half-inhibition concentration(IC50)was 65.05 mg·L-1.The LDH release rate wassignificantly increased(P<0.01)and the proportion of EdU positive cellswas significantly reduced(P<0.01)in S protein 25,50,75 and 100 mg·L-1 groups.S protein signifi-cantly reduced intracellular ATP content(P<0.01)at the concentrations of 75 and 100 mg·L-1,while significantly reduced intracellular MMP(P<0.05,P<0.01)at the concentrations of 50 and 75 mg·L-1.S protein 50 mg·L-1 increased the maximum value of basal glycolysis levels and glycolytic capacity(P<0.05,P<0.01),and S protein 25 and 50 mg·L-1 increased the maximum value of respiration capacity(P<0.05,P<0.01).SH-SY5Y cell viability was positively correlated with the intracellular ATP content and the MMP level(r2=0.9209,P=0.001;r2=0.6170,P=0.0025),and negatively correlated with the maximum level of basal glycolysis and glycolytic capacity(r2=0.5194,P=0.0285;r2=0.6664,P=0.0073),and nega-tively correlated with ATP production capacity(r2=0.8204,P=0.0008).CONCLUSIONS protein decreases the viability of SH-SY5Y cells and inhibited cell proliferation.The mechanism may be closely related to the disorder of energy metabolism.

OBJECTIVE To investigate the damage effect and mechanisms of cyclophosphamide(CTX)and its active metabolite derivative 4-hydroperoxycyclophosphamide(4-HC)to human neuroblas-toma SH-SY5Y cells.METHODS SH-SY5Y cells were treated with CTX[0(cell control),0.01,0.1,1,5,10,20,40 and 80 mmol·L-1]and 4-HC[0(cell control),0.01,0.1,1,5,10,20,40 and 80 μmol·L-1]for 48 h.Cell confluence and morphology were observed by the IncuCyte ZOOM system.Cell viability was assessed by CCK-8 assay.Lactate dehydrogenase(LDH)release was measured by LDH assay kit.SH-SY5Y cells were treated with CTX(0,1,5,10 and 20 mmol·L-1)and 4-HC(0,1,5,10 and 20 μmol·L-1)for 48 h before cell proliferation was analyzed by 5-ethynyl-2′-deoxyuridine(EdU)staining assay.Immunofluorescence was employed to assess the levels of the DNA double-strand break marker γ-H2AX and to evaluate changes in mitochondrial membrane potential.SH-SY5Y cells were treated with CTX(0,1,5 and 10 mmol·L-1)and 4-HC(0,1,5 and 10 μmol·L-1)for 48 h,and the alterations in glycolysis and oxidative phosphorylation levels were analyzed using the Seahorse XFe96 Analyzer.RESULTS Compared with the cell control group,cell confluence and cell viability were significantly reduced in the CTX and 4-HC groups(P<0.01),and the half-maximal inhibitory concentrations(IC50)for CTX and 4-HC were 4.44 mmol·L-1 and 4.78 μmol·L-1,respectively.The release rate of LDH was signif-icantly increased while the percentage of EdU+cells was significantly reduced in the CTX and 4-HC groups(P<0.01).The percentage of γ-H2AX+cells was significantly increased and mitochondrial membrane potential significantly decreased in the CTX and 4-HC group(P<0.05).Treatment with CTX and 4-HC resulted in reduced levels of maximum glycolytic capacity,glycolytic reserve,maximal respi-ration,and ATP production(P<0.05).CONCLUSION CTX and 4-HC exert significant cytotoxic effects on SH-SY5Y cells by disrupting cell membrane structure,impeding cell proliferation,and reducing cell viability.The mechanisms underlying these effects may involve intracellular DNA damage,disturbance of energy metabolism and mitochondrial dysfunction.