Objective To investigate the effect of sulforaphane(SFN)on the expression of apoptosis-related proteins(caspase-3 and caspase-9),in brain tissue of rats with acute carbon monoxide poisoning(ACOP),and to explore the molecular mechanism underlying its intervention in ACOP-induced brain injury.Methods The healthy male Sprague-Dawley(SD)rats were randomly assigned to three groups:normal control(NC)group,ACOP group,and SFN group,with 36 rats in each group.An ACOP animal model was established by exposing the rats to carbon monoxide(CO)in a hyperbaric oxygen chamber,while the rats in the NC group were allowed to breathe fresh air.The SFN group received an intraperitoneal injection of SFN 20 mg/kg within 2 hours after poisoning,once daily,until euthanasia.The NC and ACOP groups were injected with an equivalent volume of saline.Rats from each group were sacrificed on days 1,3,and 7 of the intervention to collect brain tissue,hematoxylin-eosin(HE)staining was performed to assess pathological damage in the brain tissue;Nissl staining was used to examine neuronal pathological changes;Immunohistochemistry was employed to detect the positive expression of caspase-3 and caspase-9 in the cortical region of the brain.Western blotting and quantitative reverse transcription-polymerase chain reaction(qRT-PCR)were conducted to measure the protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue.Results After CO poisoning,brain tissue damage in the ACOP group progressively worsened,with a gradual decrease in the number of Nissl bodies and a gradual increase in the number of positive cells for caspase-3 and caspase-9 in the cortical region of the brain.The protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue also gradually increased.Compared with the NC group at the same time point,the differences were statistically significant[Nissl bodies:69.33±0.94 vs.91.33±1.25;caspase-3 positive expression(A value):0.149±0.003 vs.0.113±0.004;caspase-9 positive expression(A value):0.178±0.002 vs.0.111±0.010;caspase-3 protein(caspase-3/GAPDH):1.634±0.045 vs.0.844±0.021;caspase-9 protein(caspase-9/GAPDH):1.754±0.024 vs.0.811±0.053;caspase-3 mRNA(2-ΔΔCt):1.718±0.052 vs.1;caspase-9 mRNA(2-ΔΔCt):1.722±0.066 vs.1,all P<0.05).Compared with the ACOP group at the same time point,the brain tissue damage in the SFN group improved,with a significant increase in the number of Nissl bodies(84.67±1.53 vs.69.33±0.94,P<0.05).The number of positive cells for caspase-3 and caspase-9 in the cortical region of the brain decreased significantly(A value:0.126±0.002 vs.0.149±0.003,0.127±0.002 vs.0.178±0.002,both P<0.05).The protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue were significantly reduced[caspase-3 protein(caspase-3/GAPDH):0.999±0.037 vs.1.634±0.045;caspase-9 protein(caspase-9/GAPDH):0.993±0.040 vs.1.754±0.024;caspase-3 mRNA(2-ΔΔCt):1.120±0.059 vs.1.718±0.052;caspase-9 mRNA(2-ΔΔCt):0.520±0.045 vs.1.722±0.066,all P<0.05].Conclusion SFN partially attenuated ACOP-induced brain injury in rats,potentially by downregulating both protein and mRNA expression of caspase-3 and caspase-9,thereby reducing cellular apoptosis.

Objective:To explore the neuroprotective effect and molecular mechanism of sulforaphane (SFN) on acute carbon monoxide poisoning (ACOP) in rats.Methods:A total of 135 healthy adult male Sprague-Dawley (SD) rats were randomly divided into normal control group, ACOP model group, and SFN intervention group, with 45 rats in each group. The ACOP animal model was reproduced using carbon monoxide (CO) inhalation in a hyperbaric oxygen chamber, while the normal control group was allowed to breathe fresh air freely. The rats in the SFN intervention group received intraperitoneal injection of SFN at a dose of 20 mg/kg once daily starting 2 hours after CO poisoning and continuing until euthanasia. The normal control group and the ACOP model group received equivalent volume of saline injection. Three rats from each group were sacrificed 1 day after intervention to observe the changes in the ultrastructure of neuronal mitochondria in brain tissues under transmission electron microscopy. Six rats from each group were evaluated for cognitive function using neurobehavioral test 7 days after intervention. Brain tissues of 6 rats in each group were collected 1, 3, and 7 days after intervention, and the expressions of phosphorylated-adenosine monophosphate-activated protein kinase (p-AMPK), mitofusin 2 (MFN2), and dynamin-related protein 1 (DRP1) were detected using immunohistochemistry staining and Western blotting. Linear regression analysis was performed to assess the correlations between the expression levels of above proteins.Results:In the normal control group, the rats did not exhibit any abnormalities in cognitive function or the ultrastructure of neuronal mitochondria in brain tissues. ACOP induced cognitive impairment and ultrastructural injury to neuronal mitochondria in rats. However, SFN significantly improved cognitive function in poisoned rats and mitigated the extent of neuronal mitochondrial damage. Over poisoning time, the expression levels of p-AMPK and MFN2 in the brain tissues of ACOP rats were gradually decreased, while the expression level of DRP1 was gradually increased. Compared with the normal control group, the ACOP model group showed significant differences in the expressions of p-AMPK, MFN2, and DRP1. After SFN intervention, the expression levels of above proteins were significantly reversed. Compared with the ACOP model group, the SFN intervention group exhibited a marked increase in the expressions of p-AMPK and MFN2 [p-AMPK positive expression ( A value): 0.226±0.003 vs. 0.177±0.033, p-AMPK protein (p-AMPK/GAPDH): 1.41±0.05 vs. 0.89±0.05, MFN2 positive expression ( A value): 0.241±0.004 vs. 0.165±0.007, MFN2 protein (MFN2/GAPDH): 1.33±0.04 vs. 0.79±0.03, all P < 0.05], along with a significant decrease in DRP1 expression [DRP1 positive expression ( A value): 0.103±0.002 vs. 0.214±0.011, DRP1 protein (DRP1/GAPDH): 1.00±0.03 vs. 1.50±0.03, both P < 0.05]. Linear regression analysis revealed a strong negative linear correlation between DRP1 protein expression and MFN2, p-AMPK protein expressions ( R2 values were 0.977 and 0.971, both P < 0.01), and a positive linear correlation between p-AMPK protein expression and MFN2 protein expression ( R2 = 0.985, P < 0.01). Conclusion:SFN can help maintain neuronal mitochondrial homeostasis by activating the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, thereby alleviating neuronal injury caused by ACOP.

Rosuvastatin(RVS)is an excellent drug with anti-inflammatory and lipid-lowering properties in the aca-demic and medical fields.However,this drug faces a series of challenges when used to treat atherosclerosis caused by hyperhomocysteinemia(HHcy),including high oral dosage,poor targeting,and long-term toxic side effects.In this study,we applied nanotechnology to construct a biomimetic nano-delivery system,macrophage membrane(M?m)-coated RVS-loaded Prussian blue(PB)nanoparticles(MPR NPs),for improving the bioavailability and targeting capacity of RVS,specifically to the plaque lesions associated with HHcy-induced atherosclerosis.In vitro assays demonstrated that MPR NPs effectively inhibited the Toll-like receptor 4(TLR4)/hypoxia-inducible factor-1α(HIF-1α)/nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathways,reducing pyroptosis and inflammatory response in macrophages.Additionally,MPR NPs reversed the abnormal distribution of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1)/ATP binding cassette transporter G1(ABCA1)/ATP binding cassette transporter G1(ABCG1)caused by HIF-1α,promoting cholesterol efflux and reducing lipid deposition.In vivo studies using apolipoprotein E knockout(ApoE-/-)mice confirmed the strong efficacy of MPR NPs in treating atherosclerosis with favorable bio-security,and the mechanism behind this efficacy is believed to involve the regulation of serum metabolism and the remodeling of gut microbes.These findings suggest that the synthesis of MPR NPs provides a promising nanosystem for the targeted therapy of HHcy-induced atherosclerosis.

Objective:To investigate the effects of evidence-based precision nursing in patients with sarcopenia undergoing total knee arthroplasty (TKA) .Methods:By convenience sampling, totally 41 TKA patients with sarcopenia treated in the Department of Orthopedics at The First Affiliated Hospital of Zhengzhou University from August 2021 to February 2022 were selected as the control group, and 41 TKA patients with sarcopenia treated from March 2022 to March 2023 as the observation group. Patients in the control group received conventional nursing care, while patients in the observation group received an evidence-based precision nursing plan. The differences in muscle mass, grip strength, physical activity ability, knee joint function, and quality of life between the two groups before and after the intervention were compared.Results:After the intervention, the total muscle mass, affected limb skeletal muscle mass, limb skeletal muscle mass, limb skeletal muscle mass index, and grip strength in the observation group were significantly higher than those in the control group ( P< 0.05). The scores on the Short Physical Performance Battery, Lysholm Score, and Sarcopenia Quality of Life Scale in the observation group were also significantly higher than those in the control group ( P<0.05) . Conclusions:Evidence-based precision nursing can improve postoperative muscle mass and strength, enhance physical activity ability and joint function, and improve the quality of life in TKA patients with sarcopenia.

Objective:This study collected a real-world data on survival and efficacy of gemcitabine-containing therapy in advanced breast cancer. Aimed to find the main reasons of affecting the duration of gemcitabine-base therapy in advanced breast cancer patients.Methods:Advanced breast cancer patients who received gemcitabine-base therapy from January 2017 to January 2019 were enrolled(10 hospitals). The clinicopathological data, the number of chemotherapy cycles and the reasons for treatment termination were collected and analyzed. To identify the reasons related with continuous treatment for advanced breast cancer and the factors which affect the survival and efficacy.Results:A total of 224 patients with advanced breast cancer were enrolled in this study, with a median age of 52 years (26-77 years), 55.4%(124/224) was postmenopausal. Luminal type were 83 cases, TNBC were 97 cases, and human epidermal growth factor receptor 2 (HER's-2) overexpression were 44. At the analysis, 224 patients who received the gemcitabine-based regimens were evaluated, included 5 complete reponse (CR), 77 partial response (PR), 112 stable disease (SD) and 27 progressive disease (PD). The objective response rate (ORR) was 36.6%(82/224). Seventy patients had serious adverse diseases, including leukopenia (9), neutrophilia (49), thrombocytopenia (15), and elevated transaminase (2). The median follow-up time was 41 months (26~61 months), and the median PFS was 5.6 months. The reasons of termination treatment were listed: disease progression were 90 patients; personal reasons were 51 patients; adverse drug reactions were 18 patients; completed treatment were 65 patients. It was found that progression-free survival (PFS) was significantly longer in patients receiving >6 cycles than that in patients with ≤6 cycles (8.2 months vs 5.4 months, HR=2.474, 95% CI: 1.730-3.538, P＜0.001). Conclusions:Gemcitabine-based regimen is generally well tolerated in the Chinese population and has relatively ideal clinical efficacy in the real world. The median PFS is significantly prolonged when the number of treatment cycles are appropriately increased.

Objective:This study collected a real-world data on survival and efficacy of gemcitabine-containing therapy in advanced breast cancer. Aimed to find the main reasons of affecting the duration of gemcitabine-base therapy in advanced breast cancer patients.Methods:Advanced breast cancer patients who received gemcitabine-base therapy from January 2017 to January 2019 were enrolled(10 hospitals). The clinicopathological data, the number of chemotherapy cycles and the reasons for treatment termination were collected and analyzed. To identify the reasons related with continuous treatment for advanced breast cancer and the factors which affect the survival and efficacy.Results:A total of 224 patients with advanced breast cancer were enrolled in this study, with a median age of 52 years (26-77 years), 55.4%(124/224) was postmenopausal. Luminal type were 83 cases, TNBC were 97 cases, and human epidermal growth factor receptor 2 (HER's-2) overexpression were 44. At the analysis, 224 patients who received the gemcitabine-based regimens were evaluated, included 5 complete reponse (CR), 77 partial response (PR), 112 stable disease (SD) and 27 progressive disease (PD). The objective response rate (ORR) was 36.6%(82/224). Seventy patients had serious adverse diseases, including leukopenia (9), neutrophilia (49), thrombocytopenia (15), and elevated transaminase (2). The median follow-up time was 41 months (26~61 months), and the median PFS was 5.6 months. The reasons of termination treatment were listed: disease progression were 90 patients; personal reasons were 51 patients; adverse drug reactions were 18 patients; completed treatment were 65 patients. It was found that progression-free survival (PFS) was significantly longer in patients receiving >6 cycles than that in patients with ≤6 cycles (8.2 months vs 5.4 months, HR=2.474, 95% CI: 1.730-3.538, P＜0.001). Conclusions:Gemcitabine-based regimen is generally well tolerated in the Chinese population and has relatively ideal clinical efficacy in the real world. The median PFS is significantly prolonged when the number of treatment cycles are appropriately increased.

Objective To investigate the effect of sulforaphane(SFN)on the expression of apoptosis-related proteins(caspase-3 and caspase-9),in brain tissue of rats with acute carbon monoxide poisoning(ACOP),and to explore the molecular mechanism underlying its intervention in ACOP-induced brain injury.Methods The healthy male Sprague-Dawley(SD)rats were randomly assigned to three groups:normal control(NC)group,ACOP group,and SFN group,with 36 rats in each group.An ACOP animal model was established by exposing the rats to carbon monoxide(CO)in a hyperbaric oxygen chamber,while the rats in the NC group were allowed to breathe fresh air.The SFN group received an intraperitoneal injection of SFN 20 mg/kg within 2 hours after poisoning,once daily,until euthanasia.The NC and ACOP groups were injected with an equivalent volume of saline.Rats from each group were sacrificed on days 1,3,and 7 of the intervention to collect brain tissue,hematoxylin-eosin(HE)staining was performed to assess pathological damage in the brain tissue;Nissl staining was used to examine neuronal pathological changes;Immunohistochemistry was employed to detect the positive expression of caspase-3 and caspase-9 in the cortical region of the brain.Western blotting and quantitative reverse transcription-polymerase chain reaction(qRT-PCR)were conducted to measure the protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue.Results After CO poisoning,brain tissue damage in the ACOP group progressively worsened,with a gradual decrease in the number of Nissl bodies and a gradual increase in the number of positive cells for caspase-3 and caspase-9 in the cortical region of the brain.The protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue also gradually increased.Compared with the NC group at the same time point,the differences were statistically significant[Nissl bodies:69.33±0.94 vs.91.33±1.25;caspase-3 positive expression(A value):0.149±0.003 vs.0.113±0.004;caspase-9 positive expression(A value):0.178±0.002 vs.0.111±0.010;caspase-3 protein(caspase-3/GAPDH):1.634±0.045 vs.0.844±0.021;caspase-9 protein(caspase-9/GAPDH):1.754±0.024 vs.0.811±0.053;caspase-3 mRNA(2-ΔΔCt):1.718±0.052 vs.1;caspase-9 mRNA(2-ΔΔCt):1.722±0.066 vs.1,all P<0.05).Compared with the ACOP group at the same time point,the brain tissue damage in the SFN group improved,with a significant increase in the number of Nissl bodies(84.67±1.53 vs.69.33±0.94,P<0.05).The number of positive cells for caspase-3 and caspase-9 in the cortical region of the brain decreased significantly(A value:0.126±0.002 vs.0.149±0.003,0.127±0.002 vs.0.178±0.002,both P<0.05).The protein and mRNA expression of caspase-3 and caspase-9 in the brain tissue were significantly reduced[caspase-3 protein(caspase-3/GAPDH):0.999±0.037 vs.1.634±0.045;caspase-9 protein(caspase-9/GAPDH):0.993±0.040 vs.1.754±0.024;caspase-3 mRNA(2-ΔΔCt):1.120±0.059 vs.1.718±0.052;caspase-9 mRNA(2-ΔΔCt):0.520±0.045 vs.1.722±0.066,all P<0.05].Conclusion SFN partially attenuated ACOP-induced brain injury in rats,potentially by downregulating both protein and mRNA expression of caspase-3 and caspase-9,thereby reducing cellular apoptosis.

Entinostat plus exemestane in hormone receptor-positive (HR+) advanced breast cancer (ABC) previously showed encouraging outcomes. This multicenter phase 3 trial evaluated the efficacy and safety of entinostat plus exemestane in Chinese patients with HR + ABC that relapsed/progressed after ≥1 endocrine therapy. Patients were randomized (2:1) to oral exemestane 25 mg/day plus entinostat (n = 235) or placebo (n = 119) 5 mg/week in 28-day cycles. The primary endpoint was the independent radiographic committee (IRC)-assessed progression-free survival (PFS). The median age was 52 (range, 28-75) years and 222 (62.7%) patients were postmenopausal. CDK4/6 inhibitors and fulvestrant were previously used in 23 (6.5%) and 92 (26.0%) patients, respectively. The baseline characteristics were comparable between the entinostat and placebo groups. The median PFS was 6.32 (95% CI, 5.30-9.11) and 3.72 (95% CI, 1.91-5.49) months in the entinostat and placebo groups (HR, 0.76; 95% CI, 0.58-0.98; P = 0.046), respectively. Grade ≥3 adverse events (AEs) occurred in 154 (65.5%) patients in the entinostat group versus 23 (19.3%) in the placebo group, and the most common grade ≥3 treatment-related AEs were neutropenia [103 (43.8%)], thrombocytopenia [20 (8.5%)], and leucopenia [15 (6.4%)]. Entinostat plus exemestane significantly improved PFS compared with exemestane, with generally manageable toxicities in HR + ABC (ClinicalTrials.gov #NCT03538171).

ObjectiveTo explore the effect of Xianlian Jiedu prescription （XLJDP） on the proliferation and glycolysis of human colorectal cancer HCT-116 cells and the underlying mechanism. MethodHCT-116 cells were cultured with XLJDP and then the survival rate was examined by methyl thiazolyl tetrazolium （MTT） assay. The effect on the HCT116 cell proliferation was detected by colony formation assay and 5-ethynyl-2′-deoxyuridine （EDU） incorporation assay. The amount of glucose consumed by HCT-116 cells was measured by glucose test kit， and the amount of produced lactic acid was determined by lactic acid test kit 48 h after the treatment with XLJDP. The expression of glycolysis-related proteins mammalian target of rapamycin （mTOR）， phosphorylated mTOR （p-mTOR）， glucose transporter 1 （GLUT1）， and lactate dehydrogenase （LDHA） was detected by Western blot. ResultThe half-maximal inhibitory concentration （IC₅₀） of XLJDP against HCT-116 cells was 6.82 g·L^-1. Compared with the blank group， XLJDP （1.625， 3.25， 6.50 g·L^-1） inhibited the proliferation of HCT-116 cells （P<0.05， P<0.01）. Moreover， compared with the blank group， XLJDP （1.625， 3.25， 6.50 g·L^-1） suppressed glucose uptake and lactic acid production in a dose-dependent manner （P<0.05， P<0.01）. The expression of p-mTOR/mTOR， LDHA， and GLUT1 was down-regulated by XLJDP （P<0.05， P<0.01）. ConclusionXLJDP can significantly inhibit the proliferation and the Warburg effect of glycolysis in colorectal cancer cells by regulating the mTOR signaling pathway and the down-regulating the expression of LDHA， GLUT1， and other key proteins and enzymes in glycolysis.