Cyclin-dependent kinase(CDK)4/6 inhibitors plus endocrine therapy represents the standard first-line treatment for patients with hormone receptor-positive,human epidermal growth factor receptor 2(HER2)-negative advanced breast cancer.The introduction of CDK4/6 inhibitors has significantly improved the prognosis of breast cancer patients.However,it has also brought new clinical challenges,such as disease progression and treatment resistance in many patients.Currently,there is a lack of standardized subsequent treatment options for patients whose disease progresses after CDK4/6 inhibitor combined with endocrine therapy.Endocrine therapy resistance can lead to tumor progression through estrogen receptor(ESR)-dependent or ESR-independent pathways.Novel endocrine agents have the potential to benefit breast cancer patients harboring ESR1 mutations.Patients with alterations in the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway may be particularly sensitive to targeted inhibitors of this pathway.Furthermore,newly approved or investigational antibody-drug conjugate(ADC),immunotherapy-based combinations,and novel cell cycle inhibitors have demonstrated promising anti-tumor activities.Precision medicine-based combination strategies not only expand clinical treatment options but also enable physicians to make personalized treatment decisions for patients.Biomarker-driven precision therapeutic strategies have emerged as a critical area of treatment development in the post-CDK4/6 inhibitor era.

Objective This study sought to establish a non-alcoholic fatty liver disease(NAFLD)model in Wistar-SD hypercholesterolemia(WSHc)rats induced by a high-fat diet and to reveal the pathogenesis of NAFLD in these rats through the Srebp-1 gene.Methods After 2 weeks of dietary treatment,thirty 6-week-old WSHc rats were divided into High-fat control group,HFD+AAV no load group,and HFD+AAV group,with 10 rats in each group.The HFD+AAV no load group and HFD+AAV group were intravenously injected with a vector virus and an shRNA-containing virus,respectively.WSHc rats were fed with a normal fat diet as a normal control group.Serum levels of ALT,AST,TBIL,ALP,TBA,GLU,CHOL,and TG were measured every 2 weeks.After a further 8 weeks of feeding,the rats were euthanized and livers were excised for HE staining,Oil Red O staining,Masson staining,and Sirius red staining to observe the morphology,lipid deposition,and fibrosis of the liver tissues.RT-qPCR was performed to detect the expression of lipid metabolism-related genes namely Srebp-1,Aacs,FASN and LDLR in the livers.Furthermore,hepatocytes were isolated,cultured,and divided into a normal control group and a high-fat control group.Next,expression of the Srebp-1 gene was detected by RT-qPCR.Srebp-1 knockout(KO)hepatocytes were constructed,then TG content was detected and the lipid accumulation was observed by Oil Red O staining.Results After 10 weeks of high-fat diet treatment,serum ALT(P＜0.001),ALP(P＜0.001),TBA(P＜0.05),GLU(P＜0.001),and CHOL(P＜0.001)significantly increased in WSHc rats.Abnormal lipid deposition with formation of large vacuolar lipid droplets and fibrotic lesions in livers were observed.The mRNA expression of Srebp-1 noticeably increased in WSHc rats(P＜0.001).Moreover,compared with the high-fat control group,the ALT(P＜0.05)and GLU(P＜0.01)in the HFD+AAV group decreased,and liver lipid deposition and the formation of large vacuolar lipid droplets were alleviated.Expressions of genes such as FASN(P＜0.05)and LDLR(P＜0.01)were significantly upregulated.Additionally,there was a significant increase in the expression of Srebp-1 in hepatocytes of the high-fat control group(P＜0.001),while after Srebp-1 gene knockout,cellular TG levels decreased and the degree of lipid droplet aggregation was reduced.Conclusions The Srebp-1 gene plays a regulatory role in hepatic lipid metabolism and deposition,modulating the expression of lipid metabolism-related genes in WSHc rats with NAFLD.In vitro experiments demonstrated that downregulation of Srebp-1 alleviates lipotoxic injury in hepatocytes,suggesting that the development of NAFLD in WSHc rats is closely associated with abnormally high expressions of the Srebp-1 gene.

Infectious disease animal models serve as indispensable tools for understanding the transmission patterns,pathogenesis,and anti-infective medicine.During the preparation and application of infectious animal disease models,situations inevitably arise that violate animal welfare and ethics,such as animal pain,suffering,and distress.Considering the biosafety factors,animal mortality is still used as the experimental endpoint in most experiments on infectious animals,which poses extremely high requirements for animal welfare and ethics.It is imperative to establish guiding principles or norms for the welfare and ethics of infectious animal experiments.Based on the fundamental principles of the welfare and ethics of experimental animals,this paper explores the special welfare and ethical requirements in infectious animal experiments.It emphasizes that infectious animal experiments should fully consider the balance among the scientific objectives of the research plan,animal welfare and ethics,and occupational health and safety of personnel.Based on literature research and comparative analysis of the welfare and ethical requirements of conventional animal experiments,special welfare and ethics requirements for infectious animal experiments are proposed,including personnel requirements,experimental animal selection standards,living environment management and equipment,special care and veterinary care,and humane endpoints.Personnel are required to undergo effective biosafety training,and sufficient authority should be granted to the Institutional Animal Care and Use Committee(IACUC),veterinarians,and veterinary technicians to ensure the implementation of animal welfare and ethics practices.The selection of laboratory animals should fully consider the requirements of research objectives,welfare,ethics,and biosafety,with the susceptibility and body size of laboratory animals being the key concerns in high-level biosafety laboratories.It is also clarified that the humane endpoint is an indispensable element of welfare and ethics in infectious animal experiments.Environmental enrichment and special care are necessary guarantees for achieving animal welfare and ethics.Therefore,this study can serve as a reference for relevant work.

Infectious disease animal models serve as indispensable tools for understanding the transmission patterns,pathogenesis,and anti-infective medicine.During the preparation and application of infectious animal disease models,situations inevitably arise that violate animal welfare and ethics,such as animal pain,suffering,and distress.Considering the biosafety factors,animal mortality is still used as the experimental endpoint in most experiments on infectious animals,which poses extremely high requirements for animal welfare and ethics.It is imperative to establish guiding principles or norms for the welfare and ethics of infectious animal experiments.Based on the fundamental principles of the welfare and ethics of experimental animals,this paper explores the special welfare and ethical requirements in infectious animal experiments.It emphasizes that infectious animal experiments should fully consider the balance among the scientific objectives of the research plan,animal welfare and ethics,and occupational health and safety of personnel.Based on literature research and comparative analysis of the welfare and ethical requirements of conventional animal experiments,special welfare and ethics requirements for infectious animal experiments are proposed,including personnel requirements,experimental animal selection standards,living environment management and equipment,special care and veterinary care,and humane endpoints.Personnel are required to undergo effective biosafety training,and sufficient authority should be granted to the Institutional Animal Care and Use Committee(IACUC),veterinarians,and veterinary technicians to ensure the implementation of animal welfare and ethics practices.The selection of laboratory animals should fully consider the requirements of research objectives,welfare,ethics,and biosafety,with the susceptibility and body size of laboratory animals being the key concerns in high-level biosafety laboratories.It is also clarified that the humane endpoint is an indispensable element of welfare and ethics in infectious animal experiments.Environmental enrichment and special care are necessary guarantees for achieving animal welfare and ethics.Therefore,this study can serve as a reference for relevant work.

Cyclin-dependent kinase(CDK)4/6 inhibitors plus endocrine therapy represents the standard first-line treatment for patients with hormone receptor-positive,human epidermal growth factor receptor 2(HER2)-negative advanced breast cancer.The introduction of CDK4/6 inhibitors has significantly improved the prognosis of breast cancer patients.However,it has also brought new clinical challenges,such as disease progression and treatment resistance in many patients.Currently,there is a lack of standardized subsequent treatment options for patients whose disease progresses after CDK4/6 inhibitor combined with endocrine therapy.Endocrine therapy resistance can lead to tumor progression through estrogen receptor(ESR)-dependent or ESR-independent pathways.Novel endocrine agents have the potential to benefit breast cancer patients harboring ESR1 mutations.Patients with alterations in the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway may be particularly sensitive to targeted inhibitors of this pathway.Furthermore,newly approved or investigational antibody-drug conjugate(ADC),immunotherapy-based combinations,and novel cell cycle inhibitors have demonstrated promising anti-tumor activities.Precision medicine-based combination strategies not only expand clinical treatment options but also enable physicians to make personalized treatment decisions for patients.Biomarker-driven precision therapeutic strategies have emerged as a critical area of treatment development in the post-CDK4/6 inhibitor era.

Objective This study sought to establish a non-alcoholic fatty liver disease(NAFLD)model in Wistar-SD hypercholesterolemia(WSHc)rats induced by a high-fat diet and to reveal the pathogenesis of NAFLD in these rats through the Srebp-1 gene.Methods After 2 weeks of dietary treatment,thirty 6-week-old WSHc rats were divided into High-fat control group,HFD+AAV no load group,and HFD+AAV group,with 10 rats in each group.The HFD+AAV no load group and HFD+AAV group were intravenously injected with a vector virus and an shRNA-containing virus,respectively.WSHc rats were fed with a normal fat diet as a normal control group.Serum levels of ALT,AST,TBIL,ALP,TBA,GLU,CHOL,and TG were measured every 2 weeks.After a further 8 weeks of feeding,the rats were euthanized and livers were excised for HE staining,Oil Red O staining,Masson staining,and Sirius red staining to observe the morphology,lipid deposition,and fibrosis of the liver tissues.RT-qPCR was performed to detect the expression of lipid metabolism-related genes namely Srebp-1,Aacs,FASN and LDLR in the livers.Furthermore,hepatocytes were isolated,cultured,and divided into a normal control group and a high-fat control group.Next,expression of the Srebp-1 gene was detected by RT-qPCR.Srebp-1 knockout(KO)hepatocytes were constructed,then TG content was detected and the lipid accumulation was observed by Oil Red O staining.Results After 10 weeks of high-fat diet treatment,serum ALT(P＜0.001),ALP(P＜0.001),TBA(P＜0.05),GLU(P＜0.001),and CHOL(P＜0.001)significantly increased in WSHc rats.Abnormal lipid deposition with formation of large vacuolar lipid droplets and fibrotic lesions in livers were observed.The mRNA expression of Srebp-1 noticeably increased in WSHc rats(P＜0.001).Moreover,compared with the high-fat control group,the ALT(P＜0.05)and GLU(P＜0.01)in the HFD+AAV group decreased,and liver lipid deposition and the formation of large vacuolar lipid droplets were alleviated.Expressions of genes such as FASN(P＜0.05)and LDLR(P＜0.01)were significantly upregulated.Additionally,there was a significant increase in the expression of Srebp-1 in hepatocytes of the high-fat control group(P＜0.001),while after Srebp-1 gene knockout,cellular TG levels decreased and the degree of lipid droplet aggregation was reduced.Conclusions The Srebp-1 gene plays a regulatory role in hepatic lipid metabolism and deposition,modulating the expression of lipid metabolism-related genes in WSHc rats with NAFLD.In vitro experiments demonstrated that downregulation of Srebp-1 alleviates lipotoxic injury in hepatocytes,suggesting that the development of NAFLD in WSHc rats is closely associated with abnormally high expressions of the Srebp-1 gene.

[Objective] To summarize the clinical manifestations, pathological characteristics, treatment options and prognosis of the world's first case of prostate ductal adenocarcinoma (PDA) complicated with prostate mucinous adenocarcinoma (PMA). [Methods] The clinical and follow-up data of a patient with PDA and PMA treated in Zhongnan Hospital of Wuhan University were retrospectively analyzed, and relevant literature in PubMed and CNKI databases was retrieved. [Results] The patient sought medical attention due to dysuria, frequent urination, urinary urgency and urinary pain for more than half a year, and was admitted to hospital 3 times in total.The initial diagnosis upon the first admission was benign prostatic hyperplasia complicated with prostatic abscess.After 2 months, the patient was readmitted due to worsening symptoms, received transurethral bladder neck incision+ cystoscopy+ transurethral plasma resection of the prostate, and postoperative diagnosis confirmed PDA with local PMA.Three months after surgery, the patient had bleeding.After auxiliary examinations revealed extensive metastasis, he received hormonal therapy.After 9 months, the patient died due to multiple lung metastases. [Conclusion] Early diagnosis has a significant impact on the treatment and prognosis, but there have been no previous reports of PDA combined with PMA, so the lack of specific biomarkers in the early stage has led to missed diagnosis or misdiagnoses.There is no specific treatment for PDA with PMA. Radical prostatectomy was not satisfactory in the treatment of this case.

Objective:To investigate the molecular mechanism of autophagy and apoptosis induced by ultrafine carbon black in human bronchial epithelial cells (BEAS-2B cells), and to study the intervention effect and mechanism of N-acetylcysteine (NAC) on ultrafine carbon black-induced oxidative damage in BEAS-2B cells.Methods:In March 2023, BEAS-2B cells were used as research object, an in vitro airway model exposed to ultrafine carbon black was constructed. A control group and three carbon black exposure groups (50, 100, 200 μg/ml) were set up, and the cells were treated with corresponding concentrations of ultrafine carbon black for 24 hours. In addition, the experiment was divided into control group, NAC+ control group, 100 μg/ml carbon black exposure group and NAC+ exposure group. The corresponding groups were treated with 2 mmol/L NAC for 1 h and 100 μg/ml ultrafine carbon black for 24 h, respectively. Cell viability was measured by CCK-8 assay. Intracellular reactive oxygen species (ROS) level was detected by chemical fluorescence method. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), as well as the content of malondialdehyde (MDA) were detected by colorimetry. The mRNA and protein expressions of autophagy-related genes[Atg5, Atg7, Beclin1, microtubule-associated protein light chain 3B (LC3B), p62 and lysosome-associated membrane protein 2 (LAMP2) ] and apoptosis-related genes [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase3, Caspase9 and poly (ADP-ribose) polymerase 1 (PARP1) ] were determined by fluorescence quantitative PCR and Western blot. Cell apoptosis was determined by flow cytometry.Results:Compared with the control group, the relative survival rates of BEAS-2B cells in 50, 100, 200 μg/ml carbon black exposure groups were significantly decreased, the levels of ROS and MDA were significantly increased, and the activities of SOD, GSH-Px and CAT were significantly decreased ( P<0.05). The relative survival rate, ROS and MDA levels, SOD, GSH-Px and CAT activities were significantly correlated with the exposure dose of ultrafine carbon black ( rs=-0.755, 0.826, 0.934, -0.810, -0.880, -0.840, P<0.05). Compared with the control group, the relative expression levels of Atg5, Atg7, Beclin1, LC3B, p62, LAMP2, Bax, Caspase3, Caspase9, PARP1 mRNA and Atg5, Atg7, Beclin1, LC3BⅡ, p62, LAMP2, Bax, cleaved Caspase3 (C-Caspase3), cleaved Caspase9 (C-Caspase9), cleaved PARP1 (C-PARP1) protein and the ratio of LC3BⅡ/LC3BⅠ in 50, 100 and 200 μg/ml carbon black exposure groups were significantly increased, while the relative expression levels of Bcl-2 mRNA and protein were significantly decreased ( P<0.05). The changes of the above indexes were significantly correlated with the exposure dose of carbon black ( rs=0.892, 0.879, 0.944, 0.892, 0.828, 0.880, 0.814, 0.794, 0.931, 0.918, 0.813, 0.866, 0.774, 0.695, 0.918, 0.761, 0.794, 0.944, 0.833, 0.866, 0.905, -0.886, -0.748, P<0.05). Compared with 100 μg/ml carbon black exposure group, the relative survival rate, the activities of SOD, GSH-Px and CAT in NAC+exposure group were significantly increased, while the levels of ROS and MDA were significantly decreased, and the relative expression levels of LC3B, p62 and Caspase3 mRNA and protein as well as the ratio of LC3BⅡ/LC3BⅠ were significantly decreased, and the differences were statistically significant ( P<0.05). Compared with the control group, the apoptosis rates of BEAS-2B cells in 50, 100, 200 μg/ml carbon black exposure groups were significantly increased ( P<0.05), and there was a significant positive correlation between ultrafine carbon black exposure dose and cell apoptosis rate ( rs=0.944, P<0.05). While compared with 100 μg/ml carbon black exposure group, the apoptosis rate of NAC+exposure group was significantly decreased, and the difference was statistically significant ( P<0.05) . Conclusion:Cell autophagy and apoptosis may be important pathophysiological mechanisms of ultrafine carbon black-induced oxidative damage in BEAS-2B cells. NAC can alleviate the occurrence of BEAS-2B cell damage caused by ultrafine carbon black by regulating oxidative stress and the cascading autophagy and apoptosis pathways.

Objective:To explore the effects of isosorbide mononitrate injection combined with continuous renal replacement therapy (CRRT) in patients with uremia congestive heart failure (CHF) and the effects on renal function, cardiac function, and serum parathyroid hormone (PTH), and N terminal pro-B-type natriuretic peptide (NT-proBNP) levels.Methods:A total of 120 CHF patients with uremia who were treated in the First Affiliated Hospital of Shaoyang University from May 2020 to May 2022 were selected as the research objects, and they were divided into the CRRT group and the combined group by the random number table method, with 60 cases in each group. They were treated with CRRT and isosorbide mononitrate combined with CRRT, respectively. After treatment for 1 week, the total effective rate, renal function, cardiac function, serum PTH, NT-proBNP and adverse reactions of the two groups were compared.Results:After treatment for 1 week, the total effective rate in the combined group was higher than that in the CRRT group: 98.33%(59/60) vs. 85.00%(51/60), there was statistical diffenrence ( χ2 = 5.35, P<0.05). After treatment for 1 week, the levels of serum creatinine, blood urea nitrogen and β2 microglobulin-MG in the combined group were lower than those in the CRRT group: (670.83 ± 81.80)μmol/L vs. (706.88 ± 93.27) μmol/L, (10.62 ± 2.58) mmol/L vs. (12.80 ± 3.55) mmol/L, (13.16 ± 2.98) mg/L vs. (16.00 ± 2.84) mg/L, there were statistical differences ( P<0.05). After treatment for 1 week, the left ventricular end systolic diameter and left ventricular end diastolic diameter were lower than those in the CRRT group, and left ventricular ejection fraction was higher than that in the CRRT group: (42.88 ± 4.16) mm vs. (46.37 ± 6.55) mm, (51.57 ± 8.33) mm vs. (56.42 ± 7.55) mm, (49.50 ± 6.27)% vs.(44.68 ± 5.14)%, there were statistical differences ( P<0.05). After treatment for 1 week, the levels of PTH and NT-proBNP in the combined group were lower than those in the CRRT group: (50.16 ± 7.15) ng/L vs. (53.27 ± 6.46) ng/L, 281.52 (255.46, 304.50) mmol/L vs. 362.49 (331.88, 378.42) mmol/L, there were statistical differences ( P<0.05). There was no statistical difference in adverse reactions between the two groups ( P>0.05). Conclusions:Isosorbide mononitrate combined with CRRT has a good effect on improving cardiac and renal function and reducing cardiac load in patients with uremia CHF.

Objective:To investigate the molecular mechanism of autophagy and apoptosis induced by ultrafine carbon black in human bronchial epithelial cells (BEAS-2B cells), and to study the intervention effect and mechanism of N-acetylcysteine (NAC) on ultrafine carbon black-induced oxidative damage in BEAS-2B cells.Methods:In March 2023, BEAS-2B cells were used as research object, an in vitro airway model exposed to ultrafine carbon black was constructed. A control group and three carbon black exposure groups (50, 100, 200 μg/ml) were set up, and the cells were treated with corresponding concentrations of ultrafine carbon black for 24 hours. In addition, the experiment was divided into control group, NAC+ control group, 100 μg/ml carbon black exposure group and NAC+ exposure group. The corresponding groups were treated with 2 mmol/L NAC for 1 h and 100 μg/ml ultrafine carbon black for 24 h, respectively. Cell viability was measured by CCK-8 assay. Intracellular reactive oxygen species (ROS) level was detected by chemical fluorescence method. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), as well as the content of malondialdehyde (MDA) were detected by colorimetry. The mRNA and protein expressions of autophagy-related genes[Atg5, Atg7, Beclin1, microtubule-associated protein light chain 3B (LC3B), p62 and lysosome-associated membrane protein 2 (LAMP2) ] and apoptosis-related genes [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase3, Caspase9 and poly (ADP-ribose) polymerase 1 (PARP1) ] were determined by fluorescence quantitative PCR and Western blot. Cell apoptosis was determined by flow cytometry.Results:Compared with the control group, the relative survival rates of BEAS-2B cells in 50, 100, 200 μg/ml carbon black exposure groups were significantly decreased, the levels of ROS and MDA were significantly increased, and the activities of SOD, GSH-Px and CAT were significantly decreased ( P<0.05). The relative survival rate, ROS and MDA levels, SOD, GSH-Px and CAT activities were significantly correlated with the exposure dose of ultrafine carbon black ( rs=-0.755, 0.826, 0.934, -0.810, -0.880, -0.840, P<0.05). Compared with the control group, the relative expression levels of Atg5, Atg7, Beclin1, LC3B, p62, LAMP2, Bax, Caspase3, Caspase9, PARP1 mRNA and Atg5, Atg7, Beclin1, LC3BⅡ, p62, LAMP2, Bax, cleaved Caspase3 (C-Caspase3), cleaved Caspase9 (C-Caspase9), cleaved PARP1 (C-PARP1) protein and the ratio of LC3BⅡ/LC3BⅠ in 50, 100 and 200 μg/ml carbon black exposure groups were significantly increased, while the relative expression levels of Bcl-2 mRNA and protein were significantly decreased ( P<0.05). The changes of the above indexes were significantly correlated with the exposure dose of carbon black ( rs=0.892, 0.879, 0.944, 0.892, 0.828, 0.880, 0.814, 0.794, 0.931, 0.918, 0.813, 0.866, 0.774, 0.695, 0.918, 0.761, 0.794, 0.944, 0.833, 0.866, 0.905, -0.886, -0.748, P<0.05). Compared with 100 μg/ml carbon black exposure group, the relative survival rate, the activities of SOD, GSH-Px and CAT in NAC+exposure group were significantly increased, while the levels of ROS and MDA were significantly decreased, and the relative expression levels of LC3B, p62 and Caspase3 mRNA and protein as well as the ratio of LC3BⅡ/LC3BⅠ were significantly decreased, and the differences were statistically significant ( P<0.05). Compared with the control group, the apoptosis rates of BEAS-2B cells in 50, 100, 200 μg/ml carbon black exposure groups were significantly increased ( P<0.05), and there was a significant positive correlation between ultrafine carbon black exposure dose and cell apoptosis rate ( rs=0.944, P<0.05). While compared with 100 μg/ml carbon black exposure group, the apoptosis rate of NAC+exposure group was significantly decreased, and the difference was statistically significant ( P<0.05) . Conclusion:Cell autophagy and apoptosis may be important pathophysiological mechanisms of ultrafine carbon black-induced oxidative damage in BEAS-2B cells. NAC can alleviate the occurrence of BEAS-2B cell damage caused by ultrafine carbon black by regulating oxidative stress and the cascading autophagy and apoptosis pathways.