Objective To investigate the mechanisms by which astaxanthin(AST)alleviates oxaliplatin(OXA)-induced neuropathic pain through antioxidant pathways so as to provide theoretical basis for clinical intervention.Methods Animal experiments:SD rats were divided into five groups(n=6):control group,OXA(4 mg/kg)group,OXA+Oil group,OXA+AST(5 mg/kg)group,and OXA+AST(10 mg/kg)group.Mechanical and cold pain thresholds were measured at day 0,7,14,and 21.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity in the dorsal root ganglia(DRG)were detected using the thiobarbituric acid(TBA)method and WST-1 assay,respectively.Western blotting was performed to analyze the expressions of Nrf2 and HO-1.Cell experiments:neuro-2a cells were divided into control group,OXA(50 μmol/L)group,AST(10 μmol/L)group,and OXA(50 μmol/L)+AST(10 μmol/L)group.Cells were treated with nerve growth factor(NGF,50 ng/mL)to induce growth,and morphological changes were observed under an inverted microscope.Intracellular reactive oxygen species(ROS)level and mitochondrial superoxide were measured using DCFH-DA fluorescent probe and MitoSOXTM red,respectively.Mitochondrial function was assessed by JC-1 assay.Western blotting was used to detect Nrf2 and HO-1 expressions.Results Animal experiments:① Mechanical and cold pain thresholds were reduced in OXA and OXA+Oil groups(P＜0.05),while AST significantly increased these thresholds in OXA-treated rats(P＜0.05).② SOD activity decreased while MDA content increased in the DRG of OXA-treated rats(P＜0.05).AST restored SOD activity and reduced MDA level(P＜0.05,P＜0.01).③ Western blotting showed elevated Nrf2 and HO-1 expressions in OXA group(P＞0.05),which were further upregulated by AST(P＜0.05,P＜0.01).Cell experiments:① OXA reduced the number of neurite-bearing cells and shortened the average neurite length(P＜0.05).Inverted microscopic observation revealed that AST intervention increased both parameters(P＜0.01,P＜0.001).② OXA increased intracellular and mitochondrial ROS fluorescence intensity(P＜0.05),which was attenuated by AST(P＜0.01).③ JC-1 assay revealed decreased mitochondrial membrane potential in OXA group(P＜0.01),which was partially reversed by AST(P＜0.05).④ Western blotting results showed that OXA upregulated Nrf2 and HO-1 expressions(P＜0.05,P＜0.01),and AST further enhanced their levels(P＜0.01).Conclusion AST alleviates OXA-induced neuropathic pain by promoting Nrf2/HO-1 expression,enhancing SOD activity,reducing lipid peroxidation and ROS production,and improving mitochondrial function.

Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective To investigate the mechanisms by which astaxanthin(AST)alleviates oxaliplatin(OXA)-induced neuropathic pain through antioxidant pathways so as to provide theoretical basis for clinical intervention.Methods Animal experiments:SD rats were divided into five groups(n=6):control group,OXA(4 mg/kg)group,OXA+Oil group,OXA+AST(5 mg/kg)group,and OXA+AST(10 mg/kg)group.Mechanical and cold pain thresholds were measured at day 0,7,14,and 21.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity in the dorsal root ganglia(DRG)were detected using the thiobarbituric acid(TBA)method and WST-1 assay,respectively.Western blotting was performed to analyze the expressions of Nrf2 and HO-1.Cell experiments:neuro-2a cells were divided into control group,OXA(50 μmol/L)group,AST(10 μmol/L)group,and OXA(50 μmol/L)+AST(10 μmol/L)group.Cells were treated with nerve growth factor(NGF,50 ng/mL)to induce growth,and morphological changes were observed under an inverted microscope.Intracellular reactive oxygen species(ROS)level and mitochondrial superoxide were measured using DCFH-DA fluorescent probe and MitoSOXTM red,respectively.Mitochondrial function was assessed by JC-1 assay.Western blotting was used to detect Nrf2 and HO-1 expressions.Results Animal experiments:① Mechanical and cold pain thresholds were reduced in OXA and OXA+Oil groups(P＜0.05),while AST significantly increased these thresholds in OXA-treated rats(P＜0.05).② SOD activity decreased while MDA content increased in the DRG of OXA-treated rats(P＜0.05).AST restored SOD activity and reduced MDA level(P＜0.05,P＜0.01).③ Western blotting showed elevated Nrf2 and HO-1 expressions in OXA group(P＞0.05),which were further upregulated by AST(P＜0.05,P＜0.01).Cell experiments:① OXA reduced the number of neurite-bearing cells and shortened the average neurite length(P＜0.05).Inverted microscopic observation revealed that AST intervention increased both parameters(P＜0.01,P＜0.001).② OXA increased intracellular and mitochondrial ROS fluorescence intensity(P＜0.05),which was attenuated by AST(P＜0.01).③ JC-1 assay revealed decreased mitochondrial membrane potential in OXA group(P＜0.01),which was partially reversed by AST(P＜0.05).④ Western blotting results showed that OXA upregulated Nrf2 and HO-1 expressions(P＜0.05,P＜0.01),and AST further enhanced their levels(P＜0.01).Conclusion AST alleviates OXA-induced neuropathic pain by promoting Nrf2/HO-1 expression,enhancing SOD activity,reducing lipid peroxidation and ROS production,and improving mitochondrial function.

Objective To explore the value of a clinical diagnostic model of heart failure(HF)based on disulfidptosis-related genes.Methods The differentially expressed disulfidetosis-related genes from the training set of Gene Expression Omnibus Series(GSE)57345 were obtained,and then analyzed with Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)en-richment analysis,and Metascape disease enrichment analysis.Six male C57BL/6J mice were ran-domly divided into control group(intraperitoneal injection of normal saline)and HF group(intra-peritoneal injection of isoproterenol),with 3 mice in each group.Real-time quantitative PCR was applied to detect the expression levels of key genes.Results GO enrichment analysis revealed that the differentially expressed disulfidetosis-related genes were mainly involved in platelet aggrega-tion and other aspects.KEGG showed they were significantly enriched in tight junctions,vascular smooth muscle contraction and other signaling pathways.Metascape enrichment analysis indicated that these genes were mainly related to focal glomerulosclerosis,glomerular disease,platelet dis-ease,tumor infiltration,nephrotic syndrome and other diseases.The HF group had significantly higher heart weight-to-body weight ratio,and lower ejection fraction,fractional shortening,cardiac output and stroke volume than the control group(P＜0.05,P＜0.01).The cardiac mRNA levels of BNP and MYH10 were significantly higher[1.026±0.501 vs 0.686±0.187,P=0.038;1.469(1.782,2.670)vs 0.360(0.786,1.117),P=0.000],while those of MYL6 and TLN1 was obviously lower(0.575±0.105 vs 1.000±0.202,P=0.027;0.429±0.114 vs 1.000±0.109,P=0.000)in the HF group than the control group.Conclusion Our constructed HF diagnostic model has better di-agnostic performance.

Objective To explore the value of a clinical diagnostic model of heart failure(HF)based on disulfidptosis-related genes.Methods The differentially expressed disulfidetosis-related genes from the training set of Gene Expression Omnibus Series(GSE)57345 were obtained,and then analyzed with Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)en-richment analysis,and Metascape disease enrichment analysis.Six male C57BL/6J mice were ran-domly divided into control group(intraperitoneal injection of normal saline)and HF group(intra-peritoneal injection of isoproterenol),with 3 mice in each group.Real-time quantitative PCR was applied to detect the expression levels of key genes.Results GO enrichment analysis revealed that the differentially expressed disulfidetosis-related genes were mainly involved in platelet aggrega-tion and other aspects.KEGG showed they were significantly enriched in tight junctions,vascular smooth muscle contraction and other signaling pathways.Metascape enrichment analysis indicated that these genes were mainly related to focal glomerulosclerosis,glomerular disease,platelet dis-ease,tumor infiltration,nephrotic syndrome and other diseases.The HF group had significantly higher heart weight-to-body weight ratio,and lower ejection fraction,fractional shortening,cardiac output and stroke volume than the control group(P＜0.05,P＜0.01).The cardiac mRNA levels of BNP and MYH10 were significantly higher[1.026±0.501 vs 0.686±0.187,P=0.038;1.469(1.782,2.670)vs 0.360(0.786,1.117),P=0.000],while those of MYL6 and TLN1 was obviously lower(0.575±0.105 vs 1.000±0.202,P=0.027;0.429±0.114 vs 1.000±0.109,P=0.000)in the HF group than the control group.Conclusion Our constructed HF diagnostic model has better di-agnostic performance.

Objective:To investigate the effect and safety of Guanxinning tablets combined with antiplatelet therapy on acute coronary syndrome. Methods:A total of 120 patients with acute coronary syndrome who received treatment in Shulan (Hangzhou) Hospital from January 2021 to December 2021 were included in this study. They were randomly divided into a control group and a study group ( n = 60/group). The control group was treated with antiplatelet drugs clopidogrel and aspirin. The study group was treated with clopidogrel, aspirin and Guanxinning tablets in combination. All patients were treated for 3 months. The levels of blood lipids (high-density lipoprotein cholesterol, total cholesterol, triglyceride, low-density lipoprotein cholesterol), C-reactive protein, tumor necrosis factor-α, endothelin-1, and vascular endothelial growth factor measured before and after treatment as well as the incidence of adverse reactions were compared between the two groups. Results:After treatment, serum levels of low-density lipoprotein cholesterol, total cholesterol, and triglyceride in each group were significantly decreased compared with those measured before treatment (all P < 0.05). After treatment, serum level of high-density lipoprotein cholesterol in each group was significantly increased compared with that measured before treatment ( P < 0.05). After treatment, serum levels of total cholesterol, low-density lipoprotein cholesterol, and triglyceride in the control group were (4.36 ± 1.01) mmol/L, (3.02 ± 0.28) mmol/L, and (1.98 ± 0.12) mmol/L respectively which were significantly higher than (3.98 ± 1.05) mmol/L, (2.52 ± 0.42) mmol/L, (1.58 ± 0.23) mmol/L in the study group ( t = -2.02, -7.67, -11.94, all P <0.05). Serum level of high-density lipoprotein cholesterol in the control group was significantly lower than that in the study group [(1.26 ± 0.08) mmol/L vs. (2.36 ± 0.16) mmol/L, t = 47.63, P < 0.001). After treatment, serum levels of C-reactive protein and endothelin-1 in the control group were (5.62 ± 0.56) mg/L and (86.24 ± 12.68) pg/L, respectively, which were significantly higher than (4.32 ± 0.82) mg/L and (75.26 ± 12.46) pg/L in the study group, ( t = -10.14, -4.78, both P < 0.001). After treatment, serum levels of tumor necrosis factor-α and vascular endothelial growth factor in the control group were (5.62 ± 0.56) mg/L and (76.28 ± 13.52) pg/L, which were significantly lower than (8.76 ± 1.06) mg/L and (86.32 ± 13.46) pg/L in the study group ( t = 20.23, 4.08, both P < 0.05). The incidence of adverse reactions in the study group was significantly lower than that in the control group [5.00% (3/60) vs. 8.33% (5/60), χ2 = -0.54, P > 0.05). Conclusion:Guanxinning tablets combined with antiplatelet has a remarkable therapeutic effect on acute coronary syndrome. It is highly safe and has a clinical application value.

Objective : To investigate the protective effect of adiponectin APN on myocardial injury caused by sep⁃ sis and its possible mechanism. : Methods Results : Compared with sham group , the expression of Bax , cleaved Caspase3 protein increased and the expression of Bcl⁃2 protein decreased in LPS group. Compared with LPS group , the injury in APN + LPS group was significantly alleviated , showing that the expression of Bax , cleaved caspase3 protein decreased and the expression of Bcl⁃2 protein increased. Meanwhile , compared with sham group , the expression of Cx43 increased in LPS group and decreased in APN + LPS group. Conclusion Adiponectin can attenuate LPS⁃induced cardiac injury in septic mice. The mechanism may be through the inhibition of apoptotic signal pathway and the expression of Cx43.

Objective: To investigate the role of connexin 43 ( Cx43) in acute sympathetic stress induced by phe- nylephrine (PE) overactivation of α1-adrenergic receptor ( α1-AR) in cardiomyocytes． Methods: Cardiomyocytes of H9C2 rats were randomly divided into control group，PE alone treatment group，Gap26 ( Cx43 specific inhibitor) intervention group and Gap26 alone treatment group．PE alone treatment group was treated with 50 μmol / L PE for 15 min．The Gap26 intervention group was pretreated with 0. 5 μmol / L Gap26 for 30 min，and then treated with 50 μmol / L PE for 15 min．The protein and mRNA expression levels of Cx43，NLRP3 inflammasome，interleukin-1 β (IL-1 β) ，Caspase-1，interleukin-18 (IL-18 ) were detected by Western blot and qRT-PCR. The expression and co-location of Cx43 in cardiomyocytes were observed by immunofluorescence assay，and the expression of inflamma- tory cytokines IL-1 β and IL-18 in cardiomyocytes was detected by ELISA. Results: Compared with control group， the protein and mRNA levels of Cx43，NLRP3，Caspase-1 and IL-18 in PE group increased．Compared with PE a- lone treatment group，the protein and mRNA levels of Cx43，NLRP3，Caspase-1 and IL-18 decreased after Gap26 intervention，but they were still higher than those of control group． Similarly ，immunofluorescence showed that Cx43 protein expression increased in PE alone group，while Cx43 expression was down-regulated in Gap26 inter- vention group compared with PE alone group．ELISA results showed that the expression of IL-1 β and IL-18 was sig- nificantly up-regulated in PE alone group，but down-regulated in Gap26 intervention group． Conclusion Cx43 is involved in α1-AR activation induced cardiac acute sympathetic stress by regulating NLRP3 inflammasome.

【Objective】 To study the mechanism of Apocynum leaves in the treatment of sepsis by network pharmacology method in order to explore the multi-dimensional research method of Xinjiang ethnic medicine treatment of infectious diseases and provide scientific theoretical basis for clinical medication. 【Methods】 TCMSP and literature collection were used to screen the main active components of Apocynum venetum leaf, and target prediction analysis was conducted by SwissTarget Prediction database. We used Genecards database to screen relevant targets for sepsis, used Omicshare to calculate Venn figure of the intersection targets, constructed the protein-protein interaction network diagram in the STRING database, used Ensembl for name conversion of protein targets, then entered Omicshare server for Go function and KEGG pathway enrichment of dynamic analysis. Last we used Cytoscape3.6.0 software to construct "Apocynum venetum leaf-the active ingredients-targets-Go-KEGG-sepsis" network to explore the mechanism of action of Apocynum’s resistance to sepsis. 【Results】 Kaferol, luteolin, proanthocyanidin B1 and phytosterol in Apocynum venetum leaves may treat sepsis by controlling signal transduction, regulating hormone level and anti-infection through predictive biological targets such as Akt1, VEGFA, MAPK3, EGFR, Src and PTGS2. They can play an important role through VEGF, EGFR, erbB, HIF-1, RAS and other enrichment pathways. 【Conclusion】 The active components of Apocynum venetum leaves can fight against sepsis through multiple targets and multiple pathways.

Objective:To evaluate the relationship between transforming growth factor beta-3 (TGF-β 3)/mammalian homologs of the drosophila mad gene 3 (Smad3) signaling pathway and neuronal apoptosis during reduction of focal cerebral ischemia-reperfusion (I/R) injury by isoflurane postconditioning (ISO) in rats. Methods:Sixty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 210-230 g, were randomly divided into 5 groups ( n=12 each): sham operation group (group S), cerebral I/R group (group I/R), cerebral I/R plus isoflurane postconditioning group (group I/R+ ISO), cerebral I/R plus pirfenidone group (group I/R+ P), and cerebral I/R plus isoflurane postconditioning plus pirfenidone group (group I/R+ ISO+ P). Local cerebral I/R was produced by middle cerebral artery occlusion for 1.5 h followed by 24-h reperfusion in anesthetized rats.Pirfenidone 5 μg/kg was injected into the lateral ventricle at 30 min before ischemia in I/R+ P and I/R+ ISO+ P groups.In I/R+ ISO and I/R+ ISO+ P groups, 1.5% isoflurane was inhaled for 1 h starting from the time point immediately after onset of reperfusion.Neuro-functional deficit was assessed using neurologic deficit scores (NDS) at the end of reperfusion.Then the animals were sacrificed, and brain tissues were removed for determination of the neuronal damage rate (by Nissl staining), neuronal apoptosis rate (by TUNEL), expression of TGF-β 3 (using immunofluorescence), and expression of TGF-β 3, phosphorylated Smad3 (p-Smad3), caspase-3, Bax and Bcl-2 (by Western blot). Results:Compared with group S, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3, caspase-3 and Bax was up-regulated, and the expression of p-Smad3 and Bcl-2 was down-regulated in group I/R ( P<0.05). Compared with group I/R, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly decreased, the expression of TGF-β 3, p-Smad3 and Bcl-2 was up-regulated, and the expression of caspase-3 and Bax was down-regulated in group I/R+ ISO, and the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3 and p-Smad3 was down-regulated, and the expression of caspase-3 was up-regulated in group I/R+ P ( P<0.05). Compared with group I/R+ ISO, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3, p-Smad3 and Bcl-2 was down-regulated, and the expression of caspase-3 and Bax was up-regulated in group I/R+ ISO+ P ( P<0.05). Conclusion:Isoflurane postconditioning can inhibit neuronal apoptosis by activating the TGF-β 3/Smad3 signaling pathway, thus reducing focal I/R injury in rats.