Mechanism of Modified Erchentang on COPD Inflammation Based on TNF-α/TNFR1/RIPKs Pathway / 中国实验方剂学杂志

ObjectiveBased on tumor necrosis factor alpha （TNF-α）/tumor necrosis factor receptor 1 （TNFR1）/receptor-interacting protein kinases （RIPKs） signaling pathway， this paper aims to study the effect of modified Erchentang on inflammation in rats with chronic obstructive pulmonary disease （COPD） and explore its mechanism of action. MethodA total of 60 SD rats were randomly divided into normal group， model group， high， medium， and low-dose groups （20， 10， 5 g·kg-1·d-1） of modified Erchentang， and Xiaokechuan group （3.5 mL·kg-1·d-1）， with 10 rats in each group. The COPD rat model was established by cigarette smoke combined with lipopolysaccharide （LPS）. The normal group and model group were given the same amount of normal saline for 21 days by gavage administration. The contents of TNF-α and TNFR1 in bronchoalveolar lavage fluid （BALF） of rats were detected by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expressions of RIPK1， RIPK3， and mixed lineage kinase domain-like （MLKL） in the lung tissue. The protein expressions of RIPK1， RIPK3， and MLKL in the lung tissue were detected by Western blot. The pathological changes in lung tissue were observed by hematoxylin-eosin （HE） staining. ResultCompared with the normal group， the contents of TNF-α and TNFR1 in BALF of the model group were significantly increased （P<0.01）， and the mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were significantly increased （P<0.01）. Compared with the model group， the contents of TNF-α and TNFR1 in BALF of high， medium， and low-dose groups of modified Erchentang and Xiaokechuan group were decreased （P<0.01）. The mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were decreased to different degrees （P<0.05， P<0.01）. ConclusionModified Erchentang can effectively improve the inflammatory response of lung tissue in COPD rats， and the mechanism may be by inhibiting the activation of the TNF-α/TNFR1/RIPKs signaling pathway.

Effect of Xuanfei Zhisou Prescription on IL-17 Signaling Pathway in COPD Model Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Xuanfei Zhisou prescription on the interleukin-17 （IL-17） signaling pathway in model rats with chronic obstructive pulmonary disease （COPD）. MethodA total of 60 Wistar rats were randomly divided into a blank group （10 rats） and a model group （50 rats）， and COPD model rats were established by tracheal infusion of lipopolysaccharide combined with passive fumigation. After modeling， the rats were divided into the model group， dexamethasone group， and high， medium， and low-dose Xuanfei Zhisou prescription groups （3.6， 1.8， 0.9 g·kg-1·d-1） according to the random number table. Rats in the blank group and model group were given normal saline of 10 mL·kg-1·d-1 by gavage administration， and the intervention groups of Xuanfei Zhisou prescription were given corresponding drugs. Rats in the dexamethasone group were given dexamethasone of 2.57×10-4 g·kg-1·d-1 for 28 days. The level of pulmonary function indexes in rats was measured by a pulmonary function detector. The contents of interleukin-6 （IL-6）， interleukin-8 （IL-8）， IL-17， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expressions of IL-17A， IL-17RA， nuclear factor-κB activator 1 （Act1）， tumor necrosis factor-associated factor 6 （TRAF6）， p-p38 mitogen-activated protein kinase （p-p38 MAPK）， nuclear factor-κB p65 （NF-κB p65）， and phosphorylation were detected by immunohistochemistry （IHC）. The protein expression levels of IL-17A， IL-17RA， Act1， and TRAF6 in the lung tissue were detected by Western blot. The mRNA expressions of IL-17A， IL-17RA， Act1， and TRAF6 in the lung tissue were detected by Real-time polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the serum contents of IL-6， IL-8， IL-17， IL-1β， and TNF-α in the model group were significantly increased （P<0.05）， and the flow rate and volume indexes of pulmonary function in the model group were significantly decreased （P<0.05）， while the time indexes and other indexes were significantly increased （P<0.05）. The mRNA and protein expression levels of IL-17A， IL-17RA， Act1， and TRAF6 in pulmonary tissue and the positive expressions of downstream NF-κB p65， p-NF-κB p65， and p-p38 MAPK were increased （P<0.05）. Compared with the model group， the levels of IL-6， IL-8， IL-17， IL-1β， and TNF-α in the serum of all treatment groups were decreased to varying degrees （P<0.05）， and the indexes of pulmonary function were improved to different degrees （P<0.05）. The mRNA and protein levels of IL-17A， IL-17RA， Act1， and TRAF6 and the positive expression of downstream NF-κB p65， p-NF-κB p65， and p-p38 MAPK in high and medium-dose Xuanfei Zhisou prescription groups were significantly decreased （P<0.05）. ConclusionXuanfei Zhisou prescription can effectively resist inflammation of COPD rats. The mechanism may be related to down-regulating the protein expression of IL-17A， IL-17RA， Act1， and TRAF6， inhibiting downstream NF-κB and p38 MAPK signaling pathways， and reducing the release of IL-6， IL-8， TNF-α， IL-17， and IL-1β， thus reducing the airway inflammation response.

Up-regulation of Proinflammatory Cytokines in Rostral Ventromedial Medulla Contributes to Chronic Postsurgical Pain by Promoting 5-HT Release / 中山大学学报(医学科学版)

ObjectiveTo investigate the role of proinflammatory cytokines tumor necrosis factor alpha （TNFα） and interleukin-1β （IL-1β） in rostral ventromedial medulla （RVM） in chronic postsurgical pain （CPSP） induced by skin/muscle incision and retraction （SMIR）. MethodsSD rats were randomly divided into 5 groups： ① Sham group； ② SMIR group； ③ SMIR+TNFα/IL-1β neutralizing antibody group； ④ SMIR+TNFα/IL-1β group and ⑤ SMIR+vehicle group. 50% paw mechanical withdrawal threshold （MWT） was measured by the up-down method， immunofluroscence was used to detect the TNFα and IL-1β expression and ELISA for the 5-Hydroxytryptamine （5-HT） level. ResultsSMIR elicited persistent nociceptive sensitization， upregulated TNFα and IL-1β expression in RVM neurons and astrocytes. Microinjection of TNFα or IL-1β neutralizing antibody into RVM inhibited the development of nociceptive sensitization and decreased the level of 5-HT in both RVM and spinal dorsal horn. While microinjection of recombinant TNFα or IL-1β into RVM enhanced the development of nociceptive sensitization and increased the level of 5-HT in both RVM and spinal dorsal horn. ConclusionUp-regulation of proinflammatory cytokines in RVM may contribute to SMIR induced CPSP by promoting 5-HT release.

Development and verification of a rapid detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line / 中国生物制品学杂志

@#Objective To develop and verify a rapid detection method for the biological activity of adalimumab based on U937-NF-κB-Luc cell line. Methods Using U937-NF-κB-Luc cell line as the detection cells,a method for detecting the biological activity of adalimumab was developed based on luciferase luminescence principle. The method was optimized for the concentration of tumor necrosis factor-α(TNF-α)(160 ng/mL as initial concentration,2 times serial dilution,10dilutions),the initial concentration of antibody(2 000 ng/mL,2 times serial dilution,20 dilutions),the dilution multiple of antibody(1. 5,2,3,4 times),the inoculation amount(8 × 103,2 × 104,4 × 104,6 × 104cells/well)and the incubation time(0. 5,1,2,3 h),and verified for the specificity,accuracy,precision and linear range. The relative potency of five batches of adalimumab was detected by using the optimized method and TNF-α neutralization activity method based on L929cells respectively. Results The dose-response curve of adalimumab international standard showed a typical S-type,and the data complied with the four-parameter equation y =(A-D)/[1 +(x/C)B]+ D,R2> 0. 99. The optimum concentration of TNF-α was 5 ng/mL,the initial concentration of antibody was 800 ng/mL,the dilution ratio for adalimumab was 1∶2,the inoculation amount was 2 × 104cells/well,and the induction time was 2 h. Three therapeutic monoclonal antibodies of TNF-α target,such as adalimumab,obtained good dose-response curves,while therapeutic monoclonal antibodies of other non-TNF-α targets did not show this curve. The linear regression equation of the logarithmic value of theoretical potency and the logarithmic value of the corresponding measured potency had a slope of 1. 037,and the relative bias was within the range of ± 12%. The geometric coefficient of variation(GCV)of the relative titer measured value of each sample was less than20%. The theoretical potency ranged from 64% to 156%,showing a good linear relationship with the measured values,and the fitting linear regression equation was y = 1. 037 4 x-0. 023 7,R2= 0. 998 4. There was no significant difference in the relative potency measured results of five batches of adalimumab by the two methods(t = 1. 198,P = 0. 265 1). Conclusion The developed detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line has good specificity,accuracy and precision with short time consumption(3 h),which can be used as a rapid evaluation method for the biological activity of adalimumab.

Research progress of adipokines and diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

Diabetic retinopathy(DR)is one of the common microangiopathy in diabetes and the main cause of blindness in adults. It can be seen that it is very important to find the specific target of DR prevention and treatment. Adipose tissue is not only an energy storage tissue, but also an active endocrine organ, which can release a variety of cytokines, called adipokines. Studies have shown that adipokines play an important role in the occurrence and development of DR. Adipokines can not only directly act on vascular endothelium through blood circulation, but also indirectly affect vascular endothelial function by affecting the activity of sympathetic nervous system and insulin sensitivity, which leads to dysfunction of vascular endothelial cells, increased retinal vascular permeability, neurodegeneration and neovascularization, and finally leads to the destruction of blood-retinal barrier. In recent years, the role of some new adipokines in DR has been paid more and more attention. This paper reviews the related research of several new adipokines in DR.

Hepatic and skeletal muscle tissue immunometabolism in trained rats / Inmunometabolismo del tejido hepático y muscular esquelético de ratas entrenadas / Imunometabolismo tecidual hepático e muscular esquelético de ratos treinados

ABSTRACT Introduction: Physical exercise can be an alternative for preventing and treating the harmful effects of obesity, mainly inflammatory effects on skeletal muscle and liver tissues. However, no consensus exists regarding this purpose's best physical training model. Objective: Evaluate morphological, metabolic, and inflammatory alterations in rats' skeletal and hepatic muscle tissues caused by aerobic and resistance training. Methods: 24 Wistar rats were divided into sedentary (S), aerobic (AE), and resistance training (R) groups. Blood glucose, total cholesterol, and serum triglycerides were measured periodically. After euthanasia, body mass was measured to calculate the total mass gain during the experiment. High-density lipoprotein (HDL) was measured. Adipose tissue was extracted to calculate its percentage relative to body mass and the liver, soleus, and gastrocnemius muscles for morphological analyses and concentrations of glycogen, lipids, and Tumor Necrosis Factor α (TNF-α). The Kruskall-Wallis test and Dunn's post-test were performed for statistical analysis, adopting p<0.05. Results: Both training models reduced the percentage of adipose tissue, body mass gain, and hepatic TNF-α concentration (p<0.05). AE increased serum HDL, gastrocnemius fiber diameter and reduced the fractal dimension in the soleus (p<0.05). R reduced blood glucose and serum and liver lipids, increased liver and soleus glycogen concentrations, increased gastrocnemius fiber diameter, and decreased TNF-α (p<0.05). Conclusion: Both training models reduced body mass, relative visceral adipose tissue, serum total cholesterol concentration, and liver inflammation. However, resistance training was more effective in promoting metabolic effects in the liver and skeletal muscle and reducing muscle inflammation in rats. Level of Evidence V; Expert Opinion.

RESUMEN Introducción: El ejercicio físico puede ser una alternativa para prevenir y tratar los efectos nocivos de la obesidad, principalmente los efectos inflamatorios sobre los tejidos del músculo esquelético y del hígado. Sin embargo, no existe consenso sobre cuál es el mejor modelo de entrenamiento físico para este fin. Objetivo: Evaluar las alteraciones morfológicas, metabólicas e inflamatorias del entrenamiento aeróbico y de resistencia en sobre los tejidos músculo esqueléticos y hepáticos de ratas. Métodos: 24 ratas Wistar se dividieron en grupos sedentarios (S), aeróbicos (AE) y de entrenamiento de resistencia (R). Se midieron periódicamente glucosa en sangre, colesterol total y triglicéridos. Después de la eutanasia, se midió la masa corporal para calcular la ganancia de masa total durante el experimento. Se midió la lipoproteína de alta densidad (HDL). Se extrajo tejido adiposo para calcular su porcentaje relativo a la masa corporal, así como hígado, músculos sóleo y gastrocnemio para análisis morfológicos y concentraciones de glucógeno, lípidos y Factor de Necrosis Tumoral α (TNF-α). Para el análisis estadístico fueron utilizados Kruskall-Wallis y el post-test de Dunn, adoptando p<0,05. Resultados: Ambos entrenamientos redujeron el porcentaje de tejido adiposo, masa corporal y la concentración de TNF-α hepático (p<0,05). AE aumentó el HDL sérico, el diámetro de la fibra del gastrocnemio y redujo la dimensión fractal en el sóleo (p<0,05). R redujo la glucosa en sangre y los lípidos séricos y hepáticos, aumentó las concentraciones de glucógeno hepático y sóleo, aumentó el diámetro de la fibra del gastrocnemio y disminuyó el TNF-α (p<0,05). Conclusión: Ambos modelos de entrenamiento redujeron la masa corporal, el tejido adiposo visceral relativo, la concentración sérica de colesterol total y la inflamación hepática. El entrenamiento de resistencia demostró ser más eficaz para promover los efectos metabólicos en el hígado y el músculo esquelético, además de reducir la inflamación muscular en ratas. Nivel de Evidencia V; Opinión del Especialista.

RESUMO Introdução: O exercício físico pode se apresentar como uma alternativa para prevenção e tratamento de efeitos deletérios da obesidade, principalmente efeitos inflamatórios sobre os tecidos muscular esquelético e hepático. No entanto, não há consenso quanto ao melhor modelo de treinamento físico para tal finalidade. Objetivos: Avaliar alterações morfológicas, metabólicas e inflamatórias dos treinamentos aeróbico e resistido sobre os tecidos muscular esquelético e hepático de ratos. Métodos: 24 ratos Wistar foram divididos nos grupos sedentário (S), treinamento aeróbico (AE) e resistido (R). Glicemia, colesterol total e triglicerídeos séricos foram mensurados periodicamente. Após a eutanásia, a massa corporal foi mensurada para calcular o ganho total de massa durante o experimento. A lipoproteína de alta densidade (HDL) foi dosada. O tecido adiposo foi extraído para cálculo de sua porcentagem relativa à massa corporal assim como o fígado e os músculos sóleo e gastrocnêmio para as análises morfológicas e das concentrações de glicogênio, lipídios e Fator de Necrose Tumoral α (TNF-α). Para análise estatística, foram utilizados o teste de Kruskall-Wallis e o pós-teste de Dunn, adotando-se p<0,05. Resultados: Ambos os modelos de treinamento reduziram o percentual de tecido adiposo, ganho de massa corporal e concentração hepática de TNF-α (p<0,05). AE aumentou o HDL sérico, o diâmetro das fibras do gastrocnêmio e reduziu a dimensão fractal no sóleo (p<0,05). R reduziu a glicemia e os lipídios séricos e hepáticos, aumentou a concentração de glicogênio hepático e sóleo, aumentou o diâmetro das fibras gastrocnêmicas e diminuiu o TNF-α (p<0,05). Conclusão: Ambos os modelos de treinamento reduziram a massa corporal, o tecido adiposo visceral relativo, a concentração sérica de colesterol total e a inflamação hepática. No entanto, o treinamento resistido mostrou-se mais eficaz em promover efeitos metabólicos no fígado e no músculo esquelético, além de reduzir a inflamação muscular em ratos. Nível de Evidência V; Opinião do Especialista.

New immunological aspects of peri-implantitis

ABSTRACT Objective: This study aimed to compare the levels of HIF1-α, VEGF, TNF-α, and IL-10 in the peri-implant crevicular fluid of patients with and without peri-implantitis. Methods: Forty patients, comprising 16 with and 24 without peri-implantitis were selected. Results: Patients with peri-implantitis exhibited significantly higher HIF-1α levels than those without peri-implantitis (p=0.0005). TNF-α revealed significant positive correlations with IL-10 (p=0.0008) and VEGF (p=0.0246), whereas HIF-1α and IL-10 levels (p=0.0041) demonstrated a negative and significative correlation in the peri-implantitis group. Conclusion: This study, for the first time demonstrates the balance of HIF-1α, TNFα, IL-10, and VEGF in peri-implantitis. It shows an elevated HIF-1α levels in patients with peri-implantitis, which could have stemmed from persistent inflammation- triggered hypoxia. Furthermore, the positive correlation between TNF-α and VEGF suggests intensified proinflammatory activity in peri-implantitis. Nevertheless, further studies are essential to understand these immune dynamics in peri-implantitis.

Study of Three Potential Diagnostic Biomarkers in Nasopharyngeal Carcinoma Samples from Guilan, North of Iran

Abstract Introduction Finding biomarkers for highly lethal cancers is a priority. Objective The current study was designed to understand the clinical significance of vascular endothelial growth factor (VEGF), latent membrane protein 1 (LMP1), and tumor necrosis factor-α (TNF-α) expression as the biomarkers, and evaluate their correlation with each other, in nasopharyngeal carcinoma (NPC) in the province of Guilan, North of Iran. Methods Gene expression was evaluated in 25 formalin-fixed paraffin-embedded (FFPE) blocks from cases of confirmed NPC and 20 FFPE samples of non-NPC by quantifying messenger ribonucleic acid (mRNA) and protein levels, using real-time polymerase chain reaction (PCR) and immunohistochemistry (IHC) methods, respectively. Furthermore, the correlations among the protein levels of different genes, along with the patients' demographic characteristics were assessed. Results Our findings on mRNA and protein levels demonstrated that the expression of the LMP1 gene in the NPC group was significantly elevated compared with that of the non-NPC group. In addition, the protein levels in the NPC group indicated a positive and significant correlation between LMP1 and VEGF expression. It was noted that both protein and mRNA levels showed no significant differences in the expression of TNF-α and VEGF genes between the NPC and control groups. Furthermore, there was no significant relationship between the expression of these proteins and the demographic characteristics of NPC patients. Conclusion Overall, a significant increase in LMP1 expression was observed in NPC patients, which may serve as a diagnostic biomarker for NPC. Also, LMP1 might be involved in NPC progression by inducing VEGF gene expression.

The prognostic values of the IL?10 (G1082A) and TNF?? (G308A) polymorphisms in Egyptian patients with acute lymphoblastic leukemia: A single?center study

Background: Interleukin?10 (IL?10) and tumor necrosis factor?alpha (TNF??) genes contribute to oncogenesis. We evaluated the influence of the IL?10 (G1082A) and TNF?? (G308A) polymorphisms on the prognosis and outcomes of Egyptian patients with acute lymphoblastic leukemia (ALL). Materials and Methods: We investigated 64 children and 76 adults with ALL, between 2016 and 2019, for the IL?10 (G1082A) and TNF?? (G308A) polymorphisms using allele?specific polymerase chain reaction and polymerase chain reaction杛estriction fragment length polymorphism. Survival analyses were performed using the Kaplan朚eier estimator and the log?rank test. Results: In children with ALL, the A allele of TNF?? and IL?10 polymorphisms was associated with older age (P = 0.04 and 0.03), more extramedullary disease (P = 0.02 and 0.001), positive breakpoint cluster region朅belson (BCR?ABL) rearrangement (p190; P = 0.04 and 0.001), and more relapse (P = 0.002). The IL?10 GG genotype was associated with higher overall survival in children (P = 0.026). Adults carrying the TNF?? A allele showed more extramedullary disease (P = 0.009) and relapse (P = 0.003). We also found a higher frequency of IL?10 A allele in adults with older age (P = 0.03), lower hemoglobin level (P = 0.04), positive BCR?ABL rearrangement (P = 0.001), more extramedullary disease (P = 0.001), more relapse (P = 0.002), and a longer time for the first complete remission (P = 0.003). Conclusion: A possible association exists between the A allele of IL?10 and TNF?? polymorphisms and poor prognosis in Egyptian patients with ALL, while the IL?10 GG genotype may be associated with better survival in children with ALL.

Comparative Analysis of TNF-alpha, TNF-R1, and TNF-R2 in Patients with Low-impact Fractures Due to Osteoporosis / Análise comparativa de TNF-alfa, TNF-R1 e TNF-R2 em pacientes com fraturas de baixo impacto decorrentes de osteoporose

Abstract Objective To analyze the serum levels of TNF-alpha and its TNF-R1 and TNF-R2 receptors in the blood of patients with low-impact fractures due to osteoporosis, comparing between genders and with healthy patients. Methods The present study was conducted with a blood sample of 62 patients, divided into patients with osteoporosis and healthy patients. The results were obtained using the ELISA method. Cytokine concentrations were determined based on the absorbance values obtained. Results Serum TNF-alpha levels were undetectable in female patients, while in males they were found only in one patient, with no significant difference. Similar results were found in the analyses of TNF-R1 and TNF-R2 levels, a significant increase in levels of TNF-alpha receptors in the groups of patients with osteoporosis compared with the control groupinbothsexes.There wasnosignificant difference between the sexes in the dosage of both receptors within the group with osteoporosis. There was also a positive and significant correlation in the levels of TNF-R1 and TNF-R2 only in women. Conclusion The significant increase in TNF-R1 and TNF-R2 levels in women with osteoporosis suggest that the release and expression of these receptors may be contributing differently to the development of osteoporosis in men and women.

Resumo Objetivo Analisar os níveis séricos de TNF-alfa e de seus receptores TNF-R1 e TNF-R2 no sangue de pacientes com fraturas de baixo impacto, decorrentes de osteoporose, comparando entre os sexos e com pacientes saudáveis. Métodos Oestudofoi realizadocom amostradesanguede 62 pacientes,divididos em pacientes com osteoporose e pacientes saudáveis. Os resultados foram obtidos através do método de ELISA. As concentrações de citocinas foram determinadas com base nos valores de absorbância obtidos. Resultados Os níveis séricos de TNF-alfa foram indetectáveis nos pacientes do sexo feminino, enquanto no masculino encontrou-se somente em um paciente, não havendo diferença significativa. Encontrou-se resultados semelhantes nas análises dos níveis de TNF-R1 e TNF-R2, aumento significativo nos níveis dos receptores de TNF-alfa nos grupos de pacientes com osteoporose em comparação com o grupo controle, em ambos os sexos. Não houve diferença significativa entre os sexos na dosagem de ambos os receptores dentro do grupo com osteoporose. Houve ainda correlação positiva e significativa nos níveis de TNF-R1 e TNF-R2 apenas nas mulheres. Conclusão O aumento significativo nos níveis de TNF-R1 e TNF-R2 em mulheres com osteoporose sugerem que a liberação e expressão destes receptores pode estar contribuindo de maneira distinta no desenvolvimento da osteoporose em homens e mulheres.

Biocompatibility of the Oxygenator on Pulsatile Flow by Electron Microscope

ABSTRACT Introduction: Extracorporeal perfusion flow type requires further investigation. The aim of this study is to compare the effects of pulsatile and nonpulsatile flow on oxygenator fibers that were analyzed by scanning electron microscope (SEM) and to extensively study patients' coagulation profiles, inflammatory markers, and functional blood tests. Methods: Twelve patients who had open heart surgery were randomly divided into two groups; the nonpulsatile flow (group NP, six patients) and pulsatile flow (group P, six patients) groups. Both superficial view and axial sections of the oxygenator fiber samples were examined under SEM to compare the thickness of absorbed blood proteins and amount of blood cells on the surface of oxygenators. Platelet count, coagulation profile, and inflammatory predictors were also studied from the blood samples. Results: Fibrinogen levels after cardiopulmonary bypass were significantly lower in group NP (group P, 2.57±2.78 g/L; group NP; 2.39±0.70 g/L, P=0.03). Inflammatory biomarkers such as C-reactive protein, interleukin (IL)-6, IL-12, apelin, S100β, and tumor necrosis factor alpha were comparable in both groups. Axial sections of the oxygenator fiber samples had a mean thickness of 45.2 µm and 46.5 µm in groups P and NP, respectively, and this difference is statistically significant (P=0.006). Superficial view of the fiber samples showed obviously lower platelet, leukocyte, and erythrocyte levels in group P. Conclusion: Our study demonstrated that both cellular elements and protein adsorption on oxygenator fibers are lower in the group P than in the group NP. Pulsatile perfusion has better biocompatibility on extracorporeal circulation when analyzed by SEM technique.

Oral lichen planus in a young patient: a case report with nine-year follow-up / Líquen plano oral em paciente jovem: um relato de caso com acompanhamento de nove anos

INTRODUCTION: Oral lichen planus is an inflammatory condition that affects the stratified squamous epithelium of the oral mucosa. It occurs more frequently in female patients and it is rarely observed in children, adolescents, or young adults. This study aims to report a case of oral lichen planus in a young patient with a nine-year followup. CASE REPORT: A 19-year-old man reported to the Dentistry Department with a complaint of an asymptomatic white lesion on the dorsum and left lateral border of his tongue, which had appeared a few weeks before. Two weeks later, a second lesion, very similar to the previous one, appeared on the central region of his tongue. An incisional biopsy was performed. The histological slides were stained with hematoxylin-eosin and the expression of interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) was assessed by immunohistochemistry. No pharmacological treatment was prescribed. The clinical and histopathological findings were suggestive of oral lichen planus. The IL-1ß/TNF-α expression was low. There was a spontaneous regression of the lesions after approximately one year. The nine-year follow-up showed no signs of recurrence. CONCLUSION: This case presents atypical features such as the age of the patient and the spontaneous remission of the lesions.

INTRODUÇÃO: O líquen plano oral é uma condição inflamatória que acomete o epitélio escamoso estratificado da mucosa oral. Ocorre mais frequentemente em pacientes do gênero feminino e é raramente encontrado em pacientes pediátricos ou juvenis. O objetivo do presente estudo é relatar um caso de líquen plano oral em um paciente jovem com acompanhamento de nove anos. RELATO DE CASO: Um rapaz de 19 anos procurou atendimento no Departamento de Odontologia com a queixa de uma lesão branca assintomática em região de dorso e borda lateral esquerda de sua língua, com tempo de evolução de algumas semanas. Duas semanas depois, uma segunda lesão, muito similar à primeira, apareceu na região central de sua língua. Uma biópsia incisional foi realizada. As lâminas histológicas foram coradas com hematoxilina-eosina e a expressão de interleucina-1beta (IL-1ß) e de fator de necrose tumoral alfa (TNF-α) foram avaliadas por imunohistoquímica. Nenhum tratamento farmacológico foi prescrito. Os achados clínicos e histopatológicos foram sugestivos de líquen plano oral. A expressão de IL-1ß/TNF-α foi baixa. Houve uma regressão espontânea das lesões após aproximadamente um ano. O acompanhamento de nove anos não detectou sinais de recorrência. CONCLUSÃO: Esse caso apresenta características atípicas, como a idade do paciente e a remissão espontânea das lesões.

Anti-inflammatory and antioxidant potential of Artemisia dracunculus L. aqueousextract against acetic acid induced ulcerative colitis in male Wistar rats

Ulcerative colitis (UC) is one of the two types of inflammatory bowel disease (IBD) which is increasing worldover due to modern life style. Patients with UC are prone to develop colorectal cancer. While the disease severity decides the treatment option, researchers look towards herbal medicines with anti-inflammatory properties for minimal or nil side effects. Artemisia dracunculus L., commonly called Tarragon, is a medicinal herb used in traditional Asian medicine mainly in Iran, India, Pakistan and Azerbaijan due to its special compounds. In this study, we tried to elucidate the effects of aqueous extract of tarragon on acetic acid induced ulcerative colitis (UC) in rats. Male Wistar rats were grouped into four groups of eight each viz., control; experimental control (UC was induced via luminal instillation of 4% acetic acid); and UC induced + aqueous tarragon extract (100 mg/kg) or prednisolone (2 mg/kg) orally for ten consecutive days. Tissue specimens were collected after the experimental period for evaluation of caspase-3 and cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Real-time PCR was used to monitor the levels of IL-1, IL-6 and TNF-? in colonic homogenates. Moreover, the levels of myeloperoxidase, nitric oxide and total antioxidant capacity were measured in colonic homogenates. The results showed that both treatment regimens could similarly reduce the severity of disease symptoms. Treatment with aqueous extract of tarragon caused a better improvement (P <0.05) in the levels of myeloperoxidase enzyme, and total antioxidant capacity of colonic homogenates compared to prednisolone. Nevertheless, the levels of the expression of caspase-3, and COX-2 and TNF-? were reduced in UC rats received prednisolone more than UC rats received aqueous extract of tarragon. The was no statistical difference in the levels of nitric oxide, IL-1 and IL-6 between UC rats received tarragon extract or prednisolone. Overall, these findings suggest that the aqueous extract of tarragon is a promising strategy to control ulcerative colitis. Aqueous extract can also be used as an anti-inflammatory and immune system stimulant in conditions where the immune system is damaged.

Role of PI3K/Akt/mTOR pathway-mediated macrophage autophagy in affecting the phenotype transformation of lung fibroblasts induced by silica dust exposure / 中南大学学报(医学版)

OBJECTIVES@#The phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway is one of the main signaling pathways related to autophagy. Autophagy plays a key role in the formation of silicosis fibrosis. The phenotypic transformation of lung fibroblasts into myofibroblasts is a hallmark of the transition from the inflammatory phase to the fibrotic phase in silicosis. This study aims to investigate whether the PI3K/Akt/mTOR pathway affects the phenotypic transformation of silicosis-induced lung fibroblasts into myofibroblasts via mediating macrophage autophagy.@*METHODS@#The human monocytic leukemia cell line THP-1 cells were differentiated into macrophages by treating with 100 ng/mL of phorbol ester for 24 h. Macrophages were exposed to different concentrations (0, 25, 50, 100, 200, 400 μg/mL) and different times (0, 6, 12, 24, 48 h) of SiO2 dust suspension. The survival rate of macrophages was measured by cell counting kit-8 (CCK-8) method. Enzyme linked immunosorbent assay (ELISA) was used to measure the contents of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) in the cell supernatant. The co-culture system of macrophages and HFL-1 cells was established by transwell. A blank control group, a SiO2 group, a LY294002 group, a SC79 group, a LY294002+SiO2 group, and a SC79+SiO2 group were set up in this experiment. Macrophages in the LY294002+SiO2 group were pretreated with LY294002 (PI3K inhibitor) for 18 hours, and macrophages in the SC79+SiO2 group were pretreated with SC79 (Akt activator) for 24 hours, and then exposed to SiO2 (100 μg/mL) dust suspension for 12 hours. The expression of microtubule-associated protein 1 light chain 3 (LC3) protein in macrophages was detected by the immunofluorescence method. The protein expressions of PI3K, Akt, mTOR, Beclin-1, LC3 in macrophages, and collagen III (Col III), α-smooth muscle actin (α-SMA), fibronectin (FN), matrix metalloproteinase-1 (MMP-1), tissue metalloproteinase inhibitor-1 (TIMP-1) in HFL-1 cells were measured by Western blotting.@*RESULTS@#After the macrophages were exposed to SiO2 dust suspension of different concentrations for 12 h, the survival rates of macrophages were gradually decreased with the increase of SiO2 concentration. Compared with the 0 μg/mL group, the survival rates of macrophages in the 100, 200, and 400 μg/mL groups were significantly decreased, and the concentrations of TGF-β1 and TNF-α in the cell supernatant were obviously increased (all P<0.05). When 100 μg/mL SiO2 dust suspension was applied to macrophages, the survival rates of macrophages were decreased with the prolonged exposure time. Compared with the 0 h group, the survival rates of macrophages were significantly decreased (all P<0.05), the concentrations of TGF-β1 and TNF-α in the cell supernatant were significantly increased, and the protein expression levels of Beclin-1 and LC3II were increased markedly in the 6, 12, 24, and 48 h groups (all P<0.05). Immunofluorescence results demonstrated that after exposure to SiO2 (100 μg/mL) dust for 12 h, LC3 exhibited punctate aggregation and significantly higher fluorescence intensity compared to the blank control group (P<0.05). Compared with the blank control group, the protein expressions of Col III, FN, α-SMA, MMP-1, and TIMP-1 in HFL-1 cells were up-regulated in the SiO2 group (all P<0.05). Compared with the SiO2 group, the protein expressions of PI3K, Akt, and mTOR were down-regulated and the protein expressions of LC3II and Beclin-1 were up-regulated in macrophages (all P<0.05), the contents of TNF-α and TGF-β1 in the cell supernatant were decreased (both P<0.01), and the protein expressions of Col III, FN, α-SMA, MMP-1, and TIMP-1 in HFL-1 cells were down-regulated (all P<0.05) in the LY294002+SiO2 group. Compared with the SiO2 group, the protein expressions of PI3K, Akt, and mTOR were up-regulated and the protein expressions of LC3II and Beclin-1 were down-regulated in macrophages (all P<0.05), the contents of TNF-α and TGF-β1 in the cell supernatant were increased (both P<0.01), and the protein expressions of Col III, FN, α-SMA, MMP-1, and TIMP-1 in HFL-1 cells were up-regulated (all P<0.05) in the SC79+SiO2 group.@*CONCLUSIONS@#Silica dust exposure inhibits the PI3K/Akt/mTOR pathway, increases autophagy and concentration of inflammatory factors in macrophages, and promotes the phenotype transformation of HFL-1 cells into myofibroblasts. The regulation of the PI3K/Akt/mTOR pathway can affect the autophagy induction and the concentration of inflammatory factors of macrophages by silica dust exposure, and then affect the phenotype transformation of HFL-1 cells into myofibroblasts induced by silica dust exposure.

Effect of synthetic peptide cSN50.1 on the malignant behavior of hepatocellular carcinoma HepG2 cells and its mechanism / 临床肝胆病杂志

ObjectiveTo investigate the effect of cSN50.1 on the proliferation， migration， invasion， and colony formation of HepG2 cells and its mechanism. MethodsHepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， cSN50.1 50 μmol/L， cSN50.1 70 μmol/L， and cSN50.1 90 μmol/L groups， and CCK－8 assay was used to investigate the effect of different concentrations of cSN50.1 on the proliferation of HepG2 cells and calculate half－maximal inhibitory concentration （IC50）. HepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， and cSN50.1 50 μmol/L groups， and wound healing assay， Transwell assay， and colony－forming assay were used to investigate the effect of different concentrations of cSN50.1 on the migration， invasion， and colony formation of HepG2 cells. HepG2 cells were divided into Control group， SP600125 group （an inhibitor of the AP－1 signaling pathway）， and cSN50.1 group to investigate the influence of the AP－1 signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and c－Jun protein in cytoplasm and nucleus. HepG2 cells were divided into Control group， PDTC group （an inhibitor of the NF－κB signaling pathway）， and cSN50.1 group to investigate the influence of the NF－κB signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and NF－κB protein in cytoplasm and nucleus. A one－way analysis of variance was used for comparison between multiple groups， and the SNK－q test was used for further comparison between two groups. ResultsCompared with the 0 μmol/L group， the 10 μmol/L group had no significant changes in proliferation， migration， invasion， and colony formation abilities （P >0.05）； the 30 μmol/L group had no significant change in proliferation ability （P>0.05）， but with significant reductions in migration， invasion， and colony formation abilities （P<0.05）； the 50 μmol/L group had significant reductions in proliferation， migration， invasion， and colony formation abilities （all P<0.01）； the 70 μmol/L and 90 μmol/L groups had a significant reduction in cell proliferation ability （P<0.01）， but with a cell survival rate of below 50%. Compared with the Control group， the SP600125， PDTC， and cSN50.1 groups had significant reductions in the mRNA and protein expression levels of CXCL5 and TNF－α （all P<0.05）. Compared with the Control group， the SP600125 group， the PDTC group， and the cSN50.1 group had a significant reduction in nuclear protein of c－Jun and NF－κB expression （P<0.05）； the SP600125 group and the PDTC group had a significant reduction in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）； the cSN50.1 group had a significant increase in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）. ConclusionThis study shows that cSN50.1 can inhibit the malignant behavior of hepatocellular carcinoma cells and reduce the expression of CXCL5 and TNF－α by inhibiting the nuclear import of c－Jun and NF－κB in hepatocellular carcinoma cells.

Effect of hypothermic machine perfusion on expression levels of inflammatory cytokines in rat kidney / 器官移植

Objective To evaluate the effect of hypothermic machine perfusion (HMP) on the expression levels of inflammatory cytokines in rat kidney. Methods Thirty male rats were randomly divided into the control (Control group), static cold storage group (SCS group) and HMP group, with 10 rats in each group. The velocity, intrarenal resistance and pH value of perfusion effluent were recorded during HMP. The expression levels of CXC chemokine ligand (CXCL)1, CXCL2, interferon (IFN)-β1, IFN-α4, CC chemokine ligand (CCL)2, CCL20, interleukin (IL)-17α, IL-17C and tumor necrosis factor (TNF)-α messenger RNA (mRNA) in renal tissues were evaluated by reverse transcription polymerase chain reaction (RT-PCR). Pathological changes of the kidney were observed by hematoxylin-eosin (HE) staining. Results During HMP, the velocity and intrarenal resistance remained stable, and the pH value of perfusion effluent was decreased slowly. RT-PCR showed that the relative expression levels of CXCL1, CXCL2, CCL2, CCL20, IL-17α, IL-17C and TNF-α mRNA in the SCS and HMP groups were higher compared with those in the Control group. Compared with the SCS group, the relative expression levels of CXCL1, CXCL2, CCL2, CCL20, IL-17α and TNF-α mRNA were up-regulated in the HMP group (all P<0.05). HE staining revealed that the morphology of renal cells was normal in the Control group, whereas evident epithelial necrosis, cytoplasmic vacuolation, brush border loss and epithelial shedding were observed in the SCS group. Compared with the SCS group, pathological changes in the HMP group were alleviated. Conclusions HMP may activate renal inflammation, and inhibiting the activation of inflammation during HMP is expected to further improve the effect of allograft preservation.

Polarization state and significance of macrophage in acute rejection after intestinal transplantation / 器官移植

Objective To investigate the changes of macrophage polarization during acute rejection (AR) after intestinal transplantation. Methods Six Brown Norway (BN) rats and 24 Lewis rats were divided into the sham operation group (6 Lewis rats), syngeneic transplantation group (Lewis→Lewis, 6 donors and 6 recipients) and allogeneic transplantation group (BN→Lewis, 6 donors and 6 recipients). At postoperative 7 d, the intestinal graft tissues in all groups were collected for hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Pathological manifestations and cell apoptosis were observed. The expression levels of serum cytokines related to M1 and M2 macrophage polarization were determined by enzyme-linked immunosorbent assay (ELISA). Surface markers of M1 and M2 macrophages of intestinal graft tissues in each group were co-localized and counted by immunofluorescence staining. Results HE staining and TUNEL assay showed that the intestinal epithelial morphology and structure were normal and no evident apoptotic bodies were found in the sham operation and syngeneic transplantation groups. At 7 d after transplantation, the epithelial villi structure of intestinal graft tissues was severely damaged, the number of crypts was decreased, the number of apoptotic bodies was increased, and inflammatory cells infiltrated into the whole intestinal wall, manifested with moderate to severe AR in the allogeneic transplantation group. ELISA revealed that the expression levels of serum cytokines related to M1 macrophage polarization, such as tumor necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin (IL)-12, of the recipient rats in the allogeneic transplantation group were higher than those in the sham operation and syngeneic transplantation groups. The expression levels of serum cytokines related to M2 macrophage polarization, such as IL-10 and transforming growth factor (TGF)-β, in the syngeneic transplantation group were higher compared with those in the sham operation and allogeneic transplantation group, and the differences were statistically significant (all P<0.05). Immunofluorescence staining showed that the number of M1 macrophages in the allogeneic transplantation group was higher than those in the sham operation and syngeneic transplantation groups, and the number of M2 macrophages in the syngeneic transplantation group was higher than those in the sham operation and allogeneic transplantation groups, and the differences were statistically significant (all P<0.05). Conclusions Among the allografts with AR after intestinal transplantation, a large number of macrophages, mainly M1 macrophages secreting a large number of pro-inflammatory cytokines, infiltrate into the whole intestinal wall. Regulating the direction of macrophage polarization is a potential treatment for AR after intestinal transplantation.

Effect of Zhuangyao Tongluo Formula（壮腰通络方） Containing Serum on TNF-α-mediated Vicious Circle of Pyroptosis of Nucleus Pulposus Cells / 中医杂志

ObjectiveTo explore the mechanism of Zhuangyao Tongluo Formula（壮腰通络方，ZTF） in delaying intervertebral disc degeneration. MethodsM1 macrophages were induced from THP-1 cells using LPS， IFN-γ and PMA. The induced M1 macrophages were then co-cultured with nucleus pulposus cells in a transwell system. Fetal bovine serum was used as the control serum， and the effects of different concentrations （5%， 10%， 15%， 20%） of serum from rats treated with ZTF on the activity of M1 macrophages and nucleus pulposus cells were analyzed using MTT assay. Experiment 1 was established， including the nucleus pulposus cell control group， M1 macrophage control group， nucleus pulposus cell + ZTF group， nucleus pulposus cell + TNF control group， nucleus pulposus cell + TNF + ZTF group， co-culture group， and co-culture + ZTF group. The levels of IL-1β， and IL-18 in the culture supernatant were detected using ELISA. The mRNA expression of IL-1β and IL-18 in nucleus pulposus cells was detected using qPCR. Additionally， the expression of GSDMD protein in nucleus pulposus cells was detected using cell immunofluorescence. In experiment 2， co-culture groups were constructed using TNF-α overexpression （OE） or empty vector （EV） plasmids， including co-culture group， TNF-EV + co-culture group， TNF-EV co-culture group + ZTF， co-culture + ZTF group， TNF-OE co-culture group + ZTF， and TNF-OE + co-culture group. The mRNA and protein expression of TNF-α in M1 cells in each group were detected using qPCR and WB. ResultsThe ZTF with 10% serum was selected for subsequent experiments. The results of experiment 1 showed that compared to the control group of nucleus pulposus cells， there was no statistically significant difference in the levels of IL-1β， IL-18， mRNA， and GSDMD expression in the nucleus pulposus cells + ZTF group （P＞0.05）. However， the TNF-α + co-culture group showed a significant increase in IL-1β， IL-18 levels， mRNA， and GSDMD expression （P＜0.01）. When compared to the co-culture group， the ZTF+ co-culture group showed a significant decrease in IL-1β， IL-18 levels， mRNA， and GSDMD expression （P＜0.01）. The results of experiment 2 showed that there was no statistically significant difference in TNF-α mRNA and protein expression between the empty vector plasmids + co-culture group and the co-culture group （P＞0.05）. Compared to the empty vector + co-culture group， the expression of TNF-α mRNA and protein was significantly reduced in the empty vector co-culture + ZTF group （P＜0.01）. Compared to the co-culture group and the empty vector + co-culture group， the expression of TNF-α mRNA and protein was significantly reduced in the co-culture + ZTF group （P＜0.01）. Compared to the co-culture + ZTF group， the expression of TNF-α mRNA and protein significantly increased in the overexpression vector co-culture + ZTF group （P＜0.01）. Compared to the overexpression vector co-culture + ZTF group， the expression of TNF-α mRNA and protein significantly increased in the overexpression vector co-culture group （P＜0.01）. ConclusionZTF serum can inhibit the TNF-α-induced apoptosis of nucleus pulposus cells and delay lumbar disc degeneration by reducing the expression of TNF-α in M1 macrophages.

Progress in epidermal growth factor receptor-tyrosine kinase inhibitors-associated interstitial lung disease / 中国胸心血管外科临床杂志

@#The tyrosine kinase activity of epidermal growth factor receptor (EGFR) plays a key role in tumor cell proliferation, invasion, migration, and drug resistance. Studies have shown that non-small cell lung cancer patients with somatic driver gene EGFR mutations are sensitive to and can benefit from EGFR-tyrosine kinase inhibitors (EGFR-TKIs). Nevertheless, EGFR-TKIs-related adverse events should not be ignored. Common adverse events such as diarrhea, acne-like rash and paronychia are usually manageable; although the incidence of interstitial lung disease is low, once it occurs, it is a serious threat to patients' life, and its pathogenesis is still unclear. There is very limited animal experimental and clinical research evidence on the potential mechanism of EGFR-TKIs-related interstitial lung disease in the available literature. Based on this, this article reviews the association between EGFR-TKIs and interstitial lung disease, at the same time, also discusses the research progress of EGFR-TKIs-related interstitial lung disease in combination with cytotoxic drugs or immunotherapeutic drugs and EGFR-TKIs, in order to provide a reference for the prevention and treatment of EGFR-TKIs-related interstitial lung disease in clinical practice in the future.

Change in expression of tumor necrosis factor receptor-associated factor 6 in lung tissues of rats with pulmonary hypertension / 中华麻醉学杂志

Objective:To determine the change in the expression of tumor necrosis factor receptor-associated factor 6 (TRAF6) in lung tissues of rats with pulmonary hypertension (PH).Methods:Sixteen SPF-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 200-220 g, were divided into 2 groups ( n=8 each) by the random number table method: control group (group C1) and PH group (group PH1). The model of PH was prepared by subcutaneous injection of monocrotaline. On day 28 after developing the model, the mean pulmonary arterial pressure (mPAP) was measured, and the Fulton index was calculated, and the percentage of media wall thickness of the small and medium pulmonary arteries and percentage of muscularized vessels were also calculated. The expression of TRAF6, transcription-3 (STAT3), phosphorylated STAT3 (p-STAT3) and Cyclin D1 in lung tissues was detected by Western blot, and p-STAT3/STAT3 ratio was calculated. The interaction between TRAF6 and STAT3 was determined by immunoprecipitation assay. Primarily cultured pulmonary artery smooth muscle cells of normal rats (group C2) and pulmonary artery smooth muscle cells of rats with PH (group PH2) were inoculated in 6-well plates ( n=3 each). The expression of TRAF6 mRNA was detected by quantitative polymerase chain reaction. The expression of TRAF6, STAT3, p-STAT3 and Cyclin D1 was detected by Western blot. Results:Compared with group C1, the mPAP, Fulton index, percentage of media wall thickness of the small and medium pulmonary arteries and percentage of muscularized vessels were significantly increased, the expression of TRAF6 and Cyclin D1 in lung tissues was up-regulated, the p-STAT3/STAT3 ratio was increased ( P<0.05 or 0.01), and the results of immunoprecipitation showed that TRAF6 interacted with STAT3 in group PH1. Compared with group C2, the expression of TRAF6 protein and mRNA and Cyclin D1 was significantly up-regulated, and the p-STAT3/STAT3 ratio was increased in group PH2 ( P<0.05 or 0.01). Conclusions:The expression of TRAF6 in the lung tissue is up-regulated in rats with PH, which may be related to pulmonary vascular remodeling by promoting the activation of STAT3.