Objective To analyze the relationship between the expression of ferroptosis markers in the tumor microenvironment(TME)and the prognosis of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods This prospective observational study included 100 HCC patients who received TACE treatment at Langfang Hospital of Traditional Chinese Medicine from March 2019 to June 2021 as the study subjects.The levels of 8-isoprostaglandin F2α(8-iso-PGF2α),4-hydroxy-2-nonenal(4-HNE),8-hydroxy-2'deoxyguanosine(8-OH-dG)and hepcidin in plasma were evaluated by ELISA kit at baseline(1 day before TACE),1 day after TACE and 4～8 weeks.The changes of ferroptosis related markers during TACE treatment were compared.The difference between the level of 8-iso-PGF2α,4-HNE and the baseline 1 day after TACE treatment was recorded as △8-iso-PGF2α,△4-HNE.Results Compared with the baseline,the levels of 8-iso-PGF2α and 4-HNE increased significantly and the level of hepcidin decreased significantly one day after TACE treatment,and the differences were statistically significant(t=8.03,16.29,2.92,all P<0.05).Compared with 1 day after treatment,the levels of 8-iso-PGF2α,4-HNE decreased and the level of 8-OH-dG increased at 4～8 weeks after TACE treatment,and the differences were statistically significant(t=9.12,17.17,2.63,all P<0.05).Multivariate COX analysis showed that △8-iso-PGF2α,△4-HNE and 8-iso-PGF2α 1 day after TACE treatment were independent factors affecting the overall survival after TACE(Wald χ2=5.205,13.801,6.054,all P<0.05).The survival time of patients with △4-HNE>2.01 μg/ml was significantly longer than that of patients with △4-HNE≤2.01 μg/ml(Log-rank=5.718,P=0.017),and that of patients with△8-iso-PGF2α>1.75ng/ml was sig-nificantly longer than that of patients with△8-iso-PGF2≤1.75ng/ml(Log-rank=4.163,P=0.041).Conclusion The prognosis of HCC patients who are in a state of high ferroptosis(4-HNE and 8-iso-PGF2 increased)at 1 day after TACE treatment is better,which indicate that ferroptosis mediated HCC death induced by TACE treatment.

Objective To analyze the relationship between the expression of ferroptosis markers in the tumor microenvironment(TME)and the prognosis of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods This prospective observational study included 100 HCC patients who received TACE treatment at Langfang Hospital of Traditional Chinese Medicine from March 2019 to June 2021 as the study subjects.The levels of 8-isoprostaglandin F2α(8-iso-PGF2α),4-hydroxy-2-nonenal(4-HNE),8-hydroxy-2'deoxyguanosine(8-OH-dG)and hepcidin in plasma were evaluated by ELISA kit at baseline(1 day before TACE),1 day after TACE and 4～8 weeks.The changes of ferroptosis related markers during TACE treatment were compared.The difference between the level of 8-iso-PGF2α,4-HNE and the baseline 1 day after TACE treatment was recorded as △8-iso-PGF2α,△4-HNE.Results Compared with the baseline,the levels of 8-iso-PGF2α and 4-HNE increased significantly and the level of hepcidin decreased significantly one day after TACE treatment,and the differences were statistically significant(t=8.03,16.29,2.92,all P<0.05).Compared with 1 day after treatment,the levels of 8-iso-PGF2α,4-HNE decreased and the level of 8-OH-dG increased at 4～8 weeks after TACE treatment,and the differences were statistically significant(t=9.12,17.17,2.63,all P<0.05).Multivariate COX analysis showed that △8-iso-PGF2α,△4-HNE and 8-iso-PGF2α 1 day after TACE treatment were independent factors affecting the overall survival after TACE(Wald χ2=5.205,13.801,6.054,all P<0.05).The survival time of patients with △4-HNE>2.01 μg/ml was significantly longer than that of patients with △4-HNE≤2.01 μg/ml(Log-rank=5.718,P=0.017),and that of patients with△8-iso-PGF2α>1.75ng/ml was sig-nificantly longer than that of patients with△8-iso-PGF2≤1.75ng/ml(Log-rank=4.163,P=0.041).Conclusion The prognosis of HCC patients who are in a state of high ferroptosis(4-HNE and 8-iso-PGF2 increased)at 1 day after TACE treatment is better,which indicate that ferroptosis mediated HCC death induced by TACE treatment.

Objective:To evaluate the role of nemo-like kinase (NLK) in lipopolysaccharide (LPS)-induced inflammatory responses of mouse bone marrow-derived macrophages (BMDMs) and the relationship with glycolysis.Methods:BMDMs were extracted from the wild type (WT) and NLK knockout (NLK -/-) C57BL/6 mice of either sex, aged 6-8 weeks, weighing 20-25 g, were used in this study. After induction, differentiation, maturation and purity identification, BMDMs were divided into phosphate buffer solution (PBS) control group (WT+ PBS group, NLK -/-+ PBS group, n=18) and LPS group (WT+ LPS group, NLK -/-+ LPS group, n=18) by a random number table method. LPS at a final concentration of 1 μg/ml was added to stimulate mature BMDMs for 6 h in LPS group, while the equal volume of PBS was given instead in PBS group. The cell viability was measured by Calcein/PI staining, and the expression of cellular NLK, inflammatory factors (tumor necrosis factor-alpha [TNF-α], interleukin-1 beta [IL-1β] and IL-6) and key glycolytic enzymes (hexokinase 2 [HK2], pyruvate kinase M2 (PKM2) and lactate dehydrogenase A [LDHA])mRNA was detected by real-time fluorescent quantitative polymerase chain reaction. Western blot was used to detect the expression of NLK, HK2, PKM2 and LDHA. The concentrations of TNF-α, IL-1β and IL-6 in the supernatant were determined by enzyme-linked immunosorbent assay. Glucose uptake and lactic acid production were measured by hexokinase method and colorimetric method respectively. Results:Compared with WT-type BMDMs, the expression of NLK protein and mRNA in NLK -/--type BMDMs was significantly down-regulated ( P<0.05). Compared with WT+ PBS group, the BMDM viability was significantly decreased, the expression of TNF-α, IL-1β and IL-6 was up-regulated, the concentrations of TNF-α, IL-1β and IL-6 in the supernatant were increased, the expression of NLK, HK2, PKM2 and LDHA protein and mRNA was up-regulated, and the glucose uptake and lactic acid production were increased in WT+ LPS group ( P<0.05). Compared with WT+ LPS group, the BMDM viability was significantly increased, the expression of TNF-α, IL-1β and IL-6 was down-regulated, the concentrations of TNF-α, IL-1β and IL-6 in the supernatant were decreased, the expression of NLK, HK2, PKM2 and LDHA protein and mRNA was down-regulated, and the glucose uptake and lactic acid production were decreased in NLK -/-+ LPS group ( P<0.05). Conclusions:NLK may enhance LPS-induced inflammatory responses of BMDMs by promoting glycolysis in BMDMs of mice.

Objective:To construct a framework for evaluating the quality of health economic evaluation methodology based on the pragmatic clinical trial.Methods:An evaluation framework was constructed based on existing quality evaluation tools for health economic evaluation other quality evaluation tools.The weights of each item in the framework were determined by the Delphi method,and the weighted average was calculated using the expert authority coefficient.Results:A total of 23 experts were consulted,and the expert authority coefficients were 0.88 and 0.90,respectively.The results of the Wilcoxon signed-rank test showed no statistically signifi-cant differences among the expert opinions in two rounds(P＞0.05).Finally,a framework with 3 dimensions and 8 items was estab-lished.Conclusion:The evaluation framework has high scientificity and reliability.

Diabetic retinopathy(DR)is a kind of microvascular disease caused by the long-term influence of diabetes mellitus(DM),and it is one of the main cause of global visual impairment and blindness.Its typical characteristics include microaneurysms,hard exudates,macular edema(DME)and neovascularization.Its pathogenesis is diverse,and the root cause is oxidative stress and advanced glycosylation end products.The nuclear red blood cell related factor 2(Nrf2)signa-ling pathway plays an important role in preventing various diseases.As one of the characteristics of DM,hyperglycemia will activate the generation of reactive oxygen species(ROS)in mitochondria.These oxidative stress factors activate the nucle-ar transcription factor κB pathway,becoming the main inducement of various complications of DM.This pathway will in-duce increased transcription of proinflammatory cytokines,including tumor necrosis factor-alpha,transforming growth fac-tor-beta and interleukin-1.The active ingredients of traditional Chinese medicine have significant antioxidant,anti-inflam-matory,anti-apoptotic and angiogenesis-promoting effects,and can block the progression of DR through various mecha-nisms.In this article,the research status of traditional Chinese medicine targeting the Nrf2 signaling pathway in the preven-tion and treatment of DR is reviewed to guide clinical and scientific research.

AIM To investigate the protective effect of Sanfeng Tongqiao Dropping Pills against house dust mite(HDM)-induced allergic asthma in mice.METHODS Compared to the intact BALB/c mice in the blank control group,the BALB/c mice randomly assigned into the model group,the dexamethasone group(0.67 mg/kg),and the low-dose,medium-dose,and high-dose Sanfeng Tongqiao Dropping Pills groups(15,30 and 60 mg/kg),were induced into acute allergic asthma models via weekly intraperitoneal sensitization with 0.1 mL HDM solution(0.5 mg/mL)for three weeks followed by three consecutive daily intranasal challenges with 10 μL HDM solution(2.5 mg/mL)starting in the third week.The drug administered continuously 7 days after the last excitation.The mice had their airway reactive Penh value detected,their pulmonary pathological changes observed by HE staining,their blood eosinophils(EOS)counted,their Th2 cytokines in lung tissue and serum IgE levels detected by ELISA,and their number of peripheral blood mononuclear cells(PBMC)and pulmonary Th2 cells detected by flow cytometry.Chronic allergic asthma was induced in grouped BALB/c mice through repeated intranasal challenges with 10 μL HDM solution(2.5 mg/mL)administered five times weekly for five consecutive weeks.Drug treatment continued for 14 days following the final challenge.After the final treatment,the mice had their pulmonary pathological changes observed by HE staining,and their levels of Th2 cytokines in B ALF and lung tissue and serum IgE detected by ELISA.RESULTS Compared to the blank control group,the acute allergic asthma model group exhibited increases in Penh value,EOS count and IgE level in serum,IL-4 and IL-5 levels in lung tissue(P＜0.01);obvious pulmonary inflammatory cells infiltration,and thickened airway wall;and increase in pulmonary number of Th2 cells(P＜0.01).Compared to the model group,the groups intervened with Sanfeng Tongqiao Dropping Pills demonstrated decreased Penh value,serum EOS count,IgE level and IL-5 level in lung tissue(P＜0.05,P＜0.01);reduced pulmonary inflammatory infiltration and alleviated airway wall thickening;and decreased number of pulmonary Th2 cells.Compared to the blank group,the chronic allergic asthma model group showed obvious pulmonary inflammatory infiltration and airway wall thickening;and increased EOS count and IgE level in serum,IL-4 and IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).Compared to the model group,the groups intervened with either medium-dose or high-dose Sanfeng Tongqiao Dropping Pills demonstrated reduced pulmonary inflammatory infiltration;and decreased serum EOS count,IgE level,IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).CONCLUSION Sanfeng Tongqiao Dropping Pills reduce Th2 cells in peripheral blood and lung tissue,suppress type 2 inflammation,and thereby alleviate allergic asthma.

OBJECTIVE To analyze the plasma-entering components of the ethanol extract of Yu-Ping-Feng-San by ultra-per-formance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UPLC-Q-TOF-MS)technology,so as to pro-vide a reference for the study of the pharmacological substances of Yu-Ping-Feng-San.METHODS Male BALB/c mice were ga-vaged with the ethanol extract of Yu-Ping-Feng-San(6.5 g·kg-1),and plasma samples were collected before administration(0 h)and 1,2,4,6,8,and 12 h after administration.The UPLC-Q-TOF-MS technology was used to analyze and identify the components absorbed into the plasma.RESULTS A total of 73 chemical components in the ethanol extract of Yu-Ping-Feng-San were identi-fied,and 26 plasma-entering components were identified in all plasma samples.Among the plasma-entering components,11 compo-nents were from Saposhnikoviae Radix,and 15 components were from Radix Astragali.CONCLUSION The chemical components and plasma-entering components of the ethanol extract of Yu-Ping-Feng-San were analyzed comprehensively,which provides a reference for further clarifying the pharmacological substance basis and mechanism of action of Yu-Ping-Feng-San.

Objective:To construct a framework for evaluating the quality of health economic evaluation methodology based on the pragmatic clinical trial.Methods:An evaluation framework was constructed based on existing quality evaluation tools for health economic evaluation other quality evaluation tools.The weights of each item in the framework were determined by the Delphi method,and the weighted average was calculated using the expert authority coefficient.Results:A total of 23 experts were consulted,and the expert authority coefficients were 0.88 and 0.90,respectively.The results of the Wilcoxon signed-rank test showed no statistically signifi-cant differences among the expert opinions in two rounds(P＞0.05).Finally,a framework with 3 dimensions and 8 items was estab-lished.Conclusion:The evaluation framework has high scientificity and reliability.

Diabetic retinopathy(DR)is a kind of microvascular disease caused by the long-term influence of diabetes mellitus(DM),and it is one of the main cause of global visual impairment and blindness.Its typical characteristics include microaneurysms,hard exudates,macular edema(DME)and neovascularization.Its pathogenesis is diverse,and the root cause is oxidative stress and advanced glycosylation end products.The nuclear red blood cell related factor 2(Nrf2)signa-ling pathway plays an important role in preventing various diseases.As one of the characteristics of DM,hyperglycemia will activate the generation of reactive oxygen species(ROS)in mitochondria.These oxidative stress factors activate the nucle-ar transcription factor κB pathway,becoming the main inducement of various complications of DM.This pathway will in-duce increased transcription of proinflammatory cytokines,including tumor necrosis factor-alpha,transforming growth fac-tor-beta and interleukin-1.The active ingredients of traditional Chinese medicine have significant antioxidant,anti-inflam-matory,anti-apoptotic and angiogenesis-promoting effects,and can block the progression of DR through various mecha-nisms.In this article,the research status of traditional Chinese medicine targeting the Nrf2 signaling pathway in the preven-tion and treatment of DR is reviewed to guide clinical and scientific research.