Cuproptosis, a recently identified form of regulated cell death triggered by excess intracellular copper, has emerged as a promising cytotoxic strategy for cancer therapy. However, the therapeutic efficacy of copper ionophores such as elesclomol (ES) is often hindered by cellular copper homeostasis mechanisms that limit copper influx and cuproptosis induction. To address this challenge, we developed a nanoagent utilizing outer membrane vesicle (OMV) derived from Akkermansia muciniphila (Akk) for co-delivery of antioxidant 1 copper chaperone (Atox1)-targeting siRNA and ES (siAtox1/ES@OMV) to tumors. In vitro, we demonstrated that Atox1 knockdown via siRNA significantly disrupted copper export mechanisms, resulting in elevated intracellular copper levels. Simultaneously, ES facilitated efficient copper influx and mitochondrial transport, leading to Fe-S cluster depletion, increased proteotoxic stress, and robust cuproptosis. In vivo, siAtox1/ES@OMV achieved targeted tumor delivery and induced pronounced cuproptosis. Furthermore, leveraging the immunomodulatory properties of OMVs, siAtox1/ES@OMV promoted T-cell infiltration and the activation of tumor-reactive cytotoxic T cells, enhancing tumor immune responses. The combination of siAtox1/ES-induced cuproptosis and immunogenic cell death synergistically suppressed tumor growth in both subcutaneous breast cancer and orthotopic rectal cancer mouse models. This study highlights the potential of integrating copper homeostasis disruption with a copper ionophore using an immunomodulatory OMV-based vector, offering a promising combinatorial strategy for cancer therapy.

Objective To analyze the antiviral effect of molnupiravir against influenza virus in vitro and in vivo.Methods The antiviral ability of molnupiravir against influenza virus strain H1N1 PR8 was detected in vitro and in vivo.H uman non-small cell lung cancer cell line(A549 cells)was used as an in vitro model of PR8 infection.The antiviral effects of molnupiravir on virus replication,protein synthesis,and viral particle formation were analyzed using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),Western blot(WB)assay,and plaque assay.PR8 nose-dripping infected C57BL/6 female mice were used as an in vivo infection model.The antivi-ral ability of molnupiravir was evaluated by detecting viral load,pathological changes,and immunohistochemistry in the control group,administration group,inoculation group,and inoculation-administration group.Results In vitro test results showed that molnupiravir had no significant inhibitory effect on influenza virus replication,protein syn-thesis,and virus particle formation(all P＞0.05).In vivo test results showed that compared with mice infected with H1N1 alone,the viral load in the lung tissue of mice treated with molnupiravir declined significantly,and the extent of pathological changes was milder.Immunohistochemical detection showed a significant weakening in nuclear protein(NP)antigen signal,and the expression levels of interferon(IFN)-α,interleukin(IL)-4,and IL-6 in the lungs were lower(all P＜0.01).Conclusion As a precursor with antiviral activity,molnupiravir can not exert anti-viral activity in lung-derived cells cultured in vitro,but can be transformed into an active form in the host,which significantly decreases the ability of H1N1 influenza virus to proliferate in the lungs and thereby alleviates the da-mage of virus to lung tissue.

Objective To analyze the antiviral effect of molnupiravir against influenza virus in vitro and in vivo.Methods The antiviral ability of molnupiravir against influenza virus strain H1N1 PR8 was detected in vitro and in vivo.H uman non-small cell lung cancer cell line(A549 cells)was used as an in vitro model of PR8 infection.The antiviral effects of molnupiravir on virus replication,protein synthesis,and viral particle formation were analyzed using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),Western blot(WB)assay,and plaque assay.PR8 nose-dripping infected C57BL/6 female mice were used as an in vivo infection model.The antivi-ral ability of molnupiravir was evaluated by detecting viral load,pathological changes,and immunohistochemistry in the control group,administration group,inoculation group,and inoculation-administration group.Results In vitro test results showed that molnupiravir had no significant inhibitory effect on influenza virus replication,protein syn-thesis,and virus particle formation(all P＞0.05).In vivo test results showed that compared with mice infected with H1N1 alone,the viral load in the lung tissue of mice treated with molnupiravir declined significantly,and the extent of pathological changes was milder.Immunohistochemical detection showed a significant weakening in nuclear protein(NP)antigen signal,and the expression levels of interferon(IFN)-α,interleukin(IL)-4,and IL-6 in the lungs were lower(all P＜0.01).Conclusion As a precursor with antiviral activity,molnupiravir can not exert anti-viral activity in lung-derived cells cultured in vitro,but can be transformed into an active form in the host,which significantly decreases the ability of H1N1 influenza virus to proliferate in the lungs and thereby alleviates the da-mage of virus to lung tissue.

Objective To investigate the anti-oxidative effects of alprostadil on contrast-induced nephropathy(CIN)after percuteous coronary intervention (PCI)in patients with chronic kidney disease(CKD).Methods A total of 200 patients with CKD were enrolled in our hospital.According to the random number table was divided into alprostadil 100 cases,100 cases of conventional treatment group.The levels of serum creatinine (Scr),creatinine clearance (eGFR),serum cystatin C (ScysC)and 8-hydroxy-deoxyguanine (8-OHdG)were observed before and after operation at 72 h and 7 d after operation.Results The incidence of CIN in the alprostadil group was significantly lower than that in the conventional treatment group (6% vs 12%,P<0.05).There was no significant difference in the level of Scr,eGFR,ScysC and 8-OHdG between the alprostadil group and the conventional treatment group (P>0.05).The level of Scr in the alprostadil group was significantly lower than that in the conventional treatment group at 72 h and 7 d after operation.The level of eGFR was significantly higher than that of the conventional treatment group (P<0.05).The levels of ScysC and 8-OHdG in the two groups were significantly higher than those before operation at 72 h and 7 d(P>0.05).The levels of ScysC and 8-OHdG in the alprostadil group were significantly lower than those in the conventional treatment group at 72 h and 7 d after PCI(P<0.05).Conclusion Alprostadil may improve the oxidative stress in patients with CKD and provide a preventive effect on CIN.

Objective: To study the effects of estrogen on the degenerative changes of condylar cartilage and subchondral bone in rats. Methods: 18 female SD rats aged 6 weeks were divided into control(C),unilateral anterior cross-bite(UAC) and UAC treated with estrogen(UAC + E) groups(n = 6). UAC metal prosthesis was cemented to the left incisors of maxilla and mandible of the rats in group UAC and UAC + E. Rats in UAC + E group were given pexitoneal injection of 80 μg 17β-estradiol per day. The rats in group C were untreated. Animal were sacrificed at the 4th weeks. The micro-structure of subchondral bone was observed by Micro-CT scanning. HE staining,Safranin O staining,immunohistochemical staining,TUNEL staining and TRAP staining for the observation of pathological changes of histomorphology,extracellular matrix,chondrocyte apoptosis in condylar cartilage,and osteoclasts number in subchondral bone. Results: UAC and UAC + E group showed evident osteoarthritis(OA)-like lesions. Compare with UAC group,there was a significant decrease in the expression of proteoglycan(P ＜ 0. 05),type Ⅱ collagen(P ＜ 0. 01),and a significant increase in the number of apoptotic chondrocytes(P ＜ 0. 01) in UAC + E group. As for subchondral bone,the BV/TV,Tb. Th parameters in C and UAC + E groups were significant higher than in UAC group(P ＜ 0. 01),while the BS /BV,Tb. N,Tb. Sp parameters and the osteoclasts number in C and UAC + E groups were significant fewer than in UAC groups(P ＜ 0. 01). There was no significant difference in bone ultra-parameters and osteoclasts number between C and UAC + E groups(P＞ 0. 05). Conclusion: In the model of rat TMJOA induced by unilateral anterior crossbite prosthesis,supra-physiological level of estrogen can reverse bone loss in subchondral bone,but accelerate the degradation of condylar cartilage.

Objective To explore the application value of fractional anisotropy(FA)values of magnetic resonance diffusion tensor imaging(DTI)parameters in rat rat glioma grading.Methods Sixty-seven female Wistar rats were divided into the experi-mental group(n=57)and control group(n=10)according to the random number table method.All the surviving rats were exam-ined by 3.0T DTI at 1-2 weeks(22 cases)and 3-4 weeks(35 cases)after inoculation,and the FA values of the tumor were ob-tained and compared with the pathological results.Results Among 57 tumor-loading rats,there were 18 low-grade gliomas and 39 high-grade gliomas.DTI showed that the FA value of high-grade gliomas was higher than that of the low-grade glioma,the differ-ence was statistically significant[(0.167 ± 0.035)vs.(0.147 ± 0.015),t=2.34,P<0.05].Conclusion The FA value of DTI pa-rameters can provide accurate,reliable and noninvasive imaging information for preoperative glioma grading.

Objective To investigate the correlation between morning blood pressure surge and Coronary Microvascular Dysfunction.Methods 58 cases of patients with hypertension in our hospital were given 24 h ambu-latory blood pressure monitoring(ambulatory blood pressure monitoring,ABPM).The coronary microvascular dys-function was estimated by the index of microcirculatory resistance(IMR). All cases were given biochemical test-ing,included TCH,TG,HDL-c,LDL-c and SUA.Results According to whether ABMP was arise,the patient were divided into MBPS group(n=21)and the Non-MBPS group(n=37).24 h,day,night and morning peak of systolic blood pressure were significantly higher in the morning peak group than in the average morning peak group. Multiple linear regression analysis showed that,MBPS,24 h average systolic blood pressure,day average systolic blood pressure,night average systolic blood pressure,and age were independent risk factors for coronary artery disease.Conclusion Morning blood pressure surge is closely related to severity of coronary microcirculation dysfunction. It is an independent risk factor for coronary microcirculation dysfunction. To control the blood pres-sure in patients with hypertension effectively can reduce morning peak target organ damage.

Objective To study the CYP2C19 genotype in patients with acute coronary syndrome(ACS) complicated with diabetes mellitus(DM)receiving percutaneous coronary intervension(PCI)by polymerase chain reaction(PCR),and to evaluate the efficacy of Ticagrelor in CYP2C19 gene polymorphism. Methods A total of 494 ACS patients with DM were enrolled. The patients were divided into routine treatment group and individual treatment group. Routine treatment group received 0.1 g/d aspirin/d and 75 mg/d of Clopidogrel. CYP2C19 gene polymorphism was examined in individual treatment group.(*1/*1)was classified into fast metabolic type,mutant heterozygous type(*1/*2,*1/*3)into intermediate metabolic type and mutant pure type (*2/*2,*2/*3,*3/*3) into slow metabolic type. Fast metabolic type received aspirin 0.1 g/d and Clopidogrel 75 mg/d,and intermediate and slow metabolic type received aspirin 0.1 g/d and Ticagrelor 90 mg bid for 12 months or more to observe the inci-dence of adverse cardiovascular events ,bleeding and other adverse reactions in 2 groups. Results The incidence of cardiac death ,recurrent myocardial infarction and angina pectoris ,stroke ,stent thrombosis and target vessel revascularization in routine treatment group was significantly higher than that in individual treatment group(P <0.05). There was no significant difference between two groups in terms of major bleeding and secondary bleeding (P > 0.05). The minimum bleeding rate was slightly higher in intermediate metabolic type in individual treatment group but without significantly difference when compared with that in fast metabolic type and slow metabolic type (both P>0.05). Conclusion Without elevating the risk of bleeding within 12 months,the efficacy of Ticagrelor is not affected by CYP2C19 gene polymorphism in patients with ACS complicated with DM after PCI.

Objective To compare the changes of plasma lipid indexes and coronary artery atherosclerotic plaque in TaqI B genotypes in CHD patients with impaired glucose tolerance (IGT) before and after statin therapy. Methods A total of 196 CHD with IGT and 160 controls were included. The changes of plasma lipid indexes and coronary artery atherosclerotic plaque in TaqI B genotypes were analyzed before and after Rosovastatin therapy. Sequenom Mass ARRAY platform was used to detect the CETP TaqI B SNPs. Results The genotype frequency of the B1B1, B1B2 and B2B2 in CHD with IGT group was 35.7%, 48.0% and 16.3% respectively, while in control group was 31.3%, 53.1% and 15.6% respectively. HDL-C, PA and MLA levels increased after Rosuvastatin therapy, while LDL-C, TG, TCH, Lpa, PA, EEMA and PB levels decreased. Conclusions CETP gene polymorphisms TaqI B would have association with the effects of Rosuvastatin therapy in the CHD with IGT.

OBJECTIVETo study the drug resistance of Acinetobacter baumannii (AB) producing VIM-2-type metallo-β-lactamase (MBL) isolated from burn patients of our ward against carbapenem antibiotics and its homology.

METHODSA total of 400 strains of AB (identified) were isolated from sputum, urine, blood, pus, and wound drainage. of burn patients hospitalized in our ward from September 2011 to March 2014. Drug resistance of the 400 strains of AB to 15 antibiotics, including compound sulfamothoxazole, aztreonam, etc. , was tested using the automatic microorganism identifying and drug sensitivity analyzer. Among the carbapenems-resistant AB isolates, modified Hodge test was applied to screen carbapenemase-producing strains. The carbapenemase genes of the carbapenemase-producing strains, and the mobile genetic elements class I-integron (Intl1) gene and conserved sequence (CS) of carbapenemase-producing strains carrying blaVIM-2 gene were determined with PCR and DNA sequencing. For carbapenemase-producing strains carrying blaVIM-2 gene, synergism test with imipenem-ethylene diamine tetraacetic acid (EDTA) and enhancement test with imipenem-EDTA and ceftazidime-EDTA were used to verify the MBL-producing status. Drug resistance of the VIM-2-type MBL-producing AB strains was analyzed. For VIM-2-type MBL-producing AB strains, plasmid conjugation experiment was used to explore the transfer of plasmid; outer membrane protein (OMP) CarO gene was detected by PCR. For VIM-2-type MBL-producing AB strains carrying CarO gene, the protein content of CarO was analyzed with sodium dodecyl sulfate polyacrylamide gel electro- phoresis. The repetitive consensus sequence of Enterobacteriaceae genome PCR (ERIC-PCR) was carried out for gene typing of VIM-2-type MBL-producing AB strains to analyze their homology.

RESULTS(1) The resistant rates of the 400 strains of AB against levofloxacin and compound sulfamethoxazole were low. A total of 381 carbapenems-resistant AB strains were screened, including 240 carbepenemase-producing strains. (2) Out of the 240 carbepenemase-producing strains, 18 strains were found to harbor the blaVIM-2 gene, accounting for 7.5%; 133 strains carried the blaTEM-1 gene, accounting for 55.42%; 195 strains carried the blaOXA23 gene, accounting for 81.25%; 188 strains carried the bla(armA) gene, accounting for 78.33%. (3) Eighteen carbepenemase-producing strains which carried the bla(VIM-2) gene were found to carry the Intl1 gene, showing the Intl1-VIM linkage. Simultaneously, Intl1 variable area CS showed diversity. (4) Eighteen carbepenemase-producing strains which carried the blaVIM-2 gene were verified to produce MBL. The resistant rates of the 18 strains of AB against compound sulfamethoxazole were the lowest, followed by levofloxacin and cefoperazone/sulbactam, and those against the other antibiotics were above 60.00%. (5) Through multiple joint tests, plasmid conjugation experiment positive transfer strain was not found in 18 VIM-2-type MBL-producing AB strains. (6) Nine out of the 18 VIM-2-type MBL-producing AB strains were found to carry CarO gene. The OMP CarO of VIM-2-type MBL-producing AB strains carrying CarO gene was lost or lowered in the protein content. (7) The 18 VIM-2-type MBL-producing AB strains were classified into 6 genotypes by the ERIC-PCR. There were respectively 6, 4, 3, and 1 stain (s) in genotypes A, B, C, and F, and there were 2 strains in genotypes D and E respectively.

CONCLUSIONSThe resistance mechanism of AB against carbapenems is mainly mediated by blaTEM-1, blaOXA-23, and bla(arma); meanwhile, VIM-2-type MBL-producing and lack or change in OMP CarO are attributable to carbapenems resistance of clinically isolated AB from burn wards, and the Intl1 gene may take a part in blaVIM-2 gene transmission.

Acinetobacter baumannii ; drug effects ; enzymology ; genetics ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Bacterial Proteins ; Burns ; drug therapy ; microbiology ; Carbapenems ; pharmacology ; Drug Resistance, Bacterial ; Genes, Bacterial ; Humans ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Sulbactam ; pharmacology ; beta-Lactamases ; genetics