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To study the effect of 50% ethanol extract of Bougainvillea xbuttiana on the enzymatic activity, cell via bility and cytokine production provoked by the venom of Bothrops jararaca in macro - phages. Three assays were used to study the effects of B. xbuttiana extract on the damage pro - duced by B. jararaca : Enzymatic activity was detected by measuring the proteoly tic and phos - pholipase A2; macrophages cytotoxicity was determined by the MTT method; levels of cytokine were evaluated using ELISA and a biological assay. After treatment with 300 µg/mL B. xbuttiana extract for 30 min, the proteolytic and phospholipase A2 activities of the venom were reduced to 95 and 61%, respectively. In macrophages cultures treated with B. xbuttiana extract combined with venom, the production of TNF - α, IL - 6 and IFN - γ was reduced, whereas IL - 10 was potenti - ated. Our results support the potential effect of the B. xbuttiana extract as a complementary therapy against the toxicity caused by the venom of B . jararaca snakes
Estudiar el efecto del extracto etanólico al 50% de Bougainvillea xbuttiana sobre la actividad enzimática viabilidad celular y producci ón de citoquinas provocada por el veneno de Bothrops jararaca en macrófagos Se utilizaron tres ensayos para estudiar los efectos del extracto de B. xbuttiana sobre el daño producido por B. jararaca : Se detectó actividad enzimática mediante la medición del proteolítico y fosfolipasa A2; la citotoxicidad de los macrófagos se determinó por el método MTT; Los niveles de citoquinas se evaluaron utilizando ELISA y un ensayo biológico. Después del tratamiento con 300 µg/mL de extracto de B. xbuttiana durante 30 mi n, las actividades proteolíticas y de fosfolipasa A2 del veneno se redujeron a 95 y 61%, respectivamente. En cultivos de macrófagos tratados con extracto de B. xbuttiana combinado con veneno, la producción de TNF - α, IL - 6 e IFN - γ se redujeron, mientras que IL - 10 se potenció. Nuestros resultados apoyan el efecto potencial del extracto de B. xbuttiana como terapia complementaria frente a la toxicidad provocada por el veneno de B. jararaca .
Asunto(s)
Animales , Femenino , Ratones , Extractos Vegetales/farmacología , Venenos de Crotálidos/antagonistas & inhibidores , Nyctaginaceae/química , Ensayo de Inmunoadsorción Enzimática , Supervivencia Celular/efectos de los fármacos , Citocinas , Venenos de Crotálidos/enzimología , Etanol , Fosfolipasas A2 , Macrófagos/efectos de los fármacos , Ratones Endogámicos BALB CRESUMEN
RESUMEN Introducción. El receptor de tipo Toll (TLR) que interactúe con el promastigote de Leishmania spp. determina la vía de activación celular. Objetivo. Identificar la expresión transcripcional de TLR-3, TLR-4, TLR-9, IL-12 y TNF-α en macrófagos infectados con una cepa nativa de L. braziliensis (Lbn). Métodos. La identificación de Lbn se hizo empleando qPCR para secuencias del DNA del cinetoplasto. Los macrófagos peritoneales de ratones fueron infectados con promastigotes y se midieron la producción de óxido nítrico (ON). Se cuantificaron los niveles transcripcionales para TLRs y citoquinas empleando qRT-PCR. Resultados. Lbn presentó 96% de homología con L. braziliensis. En los infectados con promastigotes se observó elevada producción de ON a las 2 h; significativa expresión transcripcional especialmente de TLR-3 y TLR-9 que se correspondió con la expresión para citoquinas. Conclusión. Lbn activó fuertemente a los macrófagos mediante los TLRs endosomales lo cual puede ser aplicado en el diseño de agonistas para tratar la enfermedad.
ABSTRACT Introduction. The Toll-like receptor (TLR) interacting with the promastigote of Leishmania spp. determines the cellular activation pathway. Objective. To determine the transcriptional expression of TLR-3, TLR-4, TLR-9, IL-12 and TNF-α in macrophages infected with a native strain of L. braziliensis (Lbn). Materials and Methods. Identification of Lbn was performed by qPCR for kinetoplast DNA sequences. Mouse peritoneal macrophages were infected with promastigotes (MI) and nitric oxide (NO) production was measured; transcript levels for TLRs and cytokines were quantified by qRT-PCR. Results. Lbn showed 96% homology to L. braziliensis. High ON production was observed in IMs at 2 h; significant transcriptional expression especially of TLR-3 and TLR-9, which corresponded with expression for cytokines. Conclusions. Lbn strongly activated macrophages via endosomal TLRs, which can be applied in the design of agonists to treat the disease.
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In this double-blind placebo-controlled randomized investigation, we assessed the tolerability of glutamine in older adults recruited from three daycare centers. The relevance of studying glutamine supplementation in elderly patients lies in its potential to provide a well-tolerated intervention. Glutamine, a crucial amino acid, plays a vital role in various physiological processes, including immune function and protein synthesis. Understanding its impact on older adults is essential, given the potential implications for their health and well-being. Participants received a daily dose of 12.4 g of oral effervescent glutamine (EGln group) or maltodextrin (placebo group) for 60 days. Fifteen patients from each group completed the study. The mean ages were 77.0±9.1 and 79.0±6.9 years for the EGln and placebo groups, respectively. We evaluated body mass index, aminogram, hemogram, plasma levels of glucose, prealbumin, albumin, urea, creatinine, uric acid, C-reactive protein, vitamin D, calcium, sodium, potassium, and the plasma activities of aspartate aminotransferase and alanine aminotransferase. Notably, we quantified a broad array of inflammatory markers and growth factors providing a holistic understanding of the potential effects of glutamine supplementation. The results demonstrated that oral glutamine did not induce significant changes in any evaluated parameters, and no adverse effects were reported. This finding suggested that the dosage of glutamine used in this study was well-tolerated and safe. This information contributes to the broader understanding of glutamine supplementation, emphasizing its safety and supporting its potential as a viable intervention for maintaining health in aging individuals.
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Considering the lack of consensus related to the impact of selective IL-6 receptor inhibition on bone remodeling and the scarcity of reports, especially on large bone defects, this study proposed to evaluate the biological impact of the selective inhibitor of interleukin-6 receptor (tocilizumab) in an experimental model of critical calvarial defect in rats. In this preclinical and in vivo study, 24 male Wistar rats were randomly divided into two groups (n=12/group): defect treated with collagen sponge (CG) and defect treated with collagen sponge associated with 2 mg/kg tocilizumab (TCZ). The defect in the parietal bone was created using an 8-mm diameter trephine drill. After 90 days, the animals were euthanized, and tissue samples (skull caps) were evaluated through micro-CT, histological, immunohistochemistry, cytokines, and RT-qPCR analyses. Tocilizumab reduced mononuclear inflammatory infiltration (P<0.05) and tumor necrosis factor (TNF)-α levels (P<0.01) and down-regulated tissue gene expression of BMP-2 (P<0.001), RUNX-2 (P<0.05), and interleukin (IL)-6 (P<0.05). Moreover, it promoted a stronger immunostaining of cathepsin and RANKL (P<0.05). Micro-CT and histological analyses revealed no impact on general bone formation (P>0.05). The bone cells (osteoblasts, osteoclasts, and osteocytes) in the defect area were similar in both groups (P>0.05). Tocilizumab reduced inflammatory cytokines, decreased osteogenic protein, and increased proteases in a critical bone defect in rats. Ninety days after the local application of tocilizumab in the cranial defect, we did not find a significant formation of bone tissue compared with a collagen sponge.
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Abstract Background: Only a fraction of patients with cutaneous lupus erythematosus (CLE) will eventually progress toward systemic disease (SLE). Objective: To find inflammatory biomarkers which could predict the progression of cutaneous lupus erythematosus (CLE) into systemic lupus erythematosus (SLE) using immunohistochemical (IHC) assays. Methods: Immunohistochemical markers for cytotoxic, inflammatory, and anti-inflammatory responses and morphometric methods were applied to routine paraffin sections of skin biopsies, taken from lesions of 59 patients with discoid lupus, subacute lupus, and lupus tumidus. For the diagnosis of SLE, patients were classified by both the American College of Rheumatology (ACR-82) and the Systemic Lupus International Collaborating Clinics (SLICC-12) systems. Results: Skin samples from CLE/SLE +patients presented higher expression of IL-1β (ARC-82: p = 0.024; SLICC-12: p = 0.0143) and a significantly higher number of cells marked with granzyme B and perforin (ARC: p = 0.0097; SLICC-12: p = 0.0148). Biopsies from CLE/SLE- individuals had higher expression of IL-17 (ARC-82: p = 0.0003; SLICC-12: p = 0.0351) and presented a positive correlation between the density of granzyme A+and FoxP3+ cells (ARC-82: p = 0.0257; SLICC-12: p = 0.0285) and CD8+ cells (ARC-82: p = 0.0075; SLICC-12: p = 0.0102), as well as between granulysin-positive and CD8+ cells (ARC-82: p = 0.0024; SLICC-12: p = 0.0116). Study limitations: Patients were evaluated at a specific point in their evolution and according to the presence or not of systemic disease. The authors cannot predict how many more, from each group, would have evolved towards SLE in the following years. Conclusions: In this cohort, immunohistochemical findings suggested that patients with a tendency to systemic disease will show strong reactivity for IL-1β, while those with purely cutaneous involvement will tend to express IL-17 more intensely.
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Objective To investigate the expressions of 12 cytokines(IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,IFN-α,IFN-γ,TNF-α)and procalcitonin in patients with infective endocarditis(IE).Methods Ten IE patients admitted to our hospital from December 2021 to December 2022 were included into the IE group,10 patients with non-infectious and non-rheumatic valvular diseases who were admitted to our hospital at the same period were randomly selected as the control group,and blood sampling of all patients were conducted at admission.The expressions of 12 cytokines and blood routine indexes were detected by flow cytometry,and the level of procalcitonin was detected by ELISA.The correlations among the expression levels of cytokines in IE patients were analyzed by Pearson method and the correlations of IL-8 level and white blood cell count with procalcitonin in IE patients were analyzed by Spearman method.Results Compared with the control group,the levels of cytokines of IL-1β,IL-2,IL-6,IL-10,TNF-α,IFN-α,IFN-γ and IL-12p70 in the IE group were significantly increased(P<0.05),the white blood cell count,neutrophil percentage and procalcitonin were significantly increased(P<0.05).There was no significant difference in the percentage of monocytes between the two groups(P>0.05).IFN-α of IE patients was positively correlated with IL-2,TNF-α,IL-1β and IL-12p70,IL-2 was positively correlated with TNF-α and IL-1β,IL-12p70 was positively correlated with IFN-γ,and procalcitonin was significantly positively correlated with IL-8 and white blood cell count,with statistically significant differences(P<0.05).Conclusion The levels of IL-1β,IL-2,IL-6,IL-10,TNF-α,IFN-α,IFN-γ,IL-12p70 and procalcitonin in IE patients are significantly higher than those in the normal population,and the detections of these indicators are of guiding significance for the early diagnosis of IE and the evaluation of the severity of the disease.
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Objective:The purpose of this study is to explore the clinical characteristics of Coronavirus Disease 2019 (COVID-19) in patients with type 2 diabetes mellitus (T2DM), and analyze the risk factors for adverse outcomes.Methods:2 052 patients diagnosed with COVID-19 who were hospitalized in Shanxi Bethune Hospital between December 1, 2022 and March 20, 2023 were included. They were divided into diabetes group ( n=70) and non-diabetes group ( n=1 982) according to the presence or absence of comorbid T2DM. The two groups were matched at 1:1 via propensity score matching. Clinical characteristics and laboratory examination results of the two groups were compared. According to the outcomes during hospitalization, the two groups were further divided into two subgroups respectively. Univariate analysis and subsequent binary Logistic regression was used to analyze the risk factors of adverse outcomes in patients with COVID-19 and type 2 diabetes. Results:After the propensity score matching, the most common comorbid condition in diabetes group and non-diabetes group was hypertension. The proportion of patients with severe or critical disease in diabetes group was higher compared with non-diabetes group. The levels of hemoglobin A1c (HbA1c), fasting blood glucose (FBG), blood urea, IL-4, IL-6, IL-10, IFN-γ and TNF-α were significantly higher in the diabetes group ( P<0.05). Logistic regression analysis within the diabetes group showed that hypertension ( OR=3.640, 95% CI: 3.156 to 4.290), FBG>11 mmol/L ( OR=3.283, 95% CI: 1.416 to 7.611), HbA1c>10% ( OR=2.718, 95% CI: 1.024 to 7.213) were independent risk factors for adverse outcomes in patients with COVID-19 and type 2 diabetes(all P<0.05). Conclusions:Compared with the non-diabetes group, patients with COVID-19 and T2DM have worse inflammatory response and higher levels of inflammatory cytokines. The elevated levels of FBG and HbA1c are related to the adverse outcome in patients with COVID-19 and T2DM.
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Autoimmune disease is a kind of heterogeneous disease caused by the loss of immune tolerance to autoantigens.In recent years,antibody drug has become an important choice for the treatment of autoimmune diseases.Monoclonal antibodies are limited by their large relative molecular mass and difficult to penetrate solid tissues,while nanobodies,the smallest antibody fragment known to date with intact antigen recognition,have gained much attention in the treatment of autoimmune diseases due to their ease of modifi-cation,high penetration,and ability to target more cryptic epitopes.The review focuses on the application of nanobodies in the treat-ment of diseases such as rheumatoid arthritis,acquired thrombotic thrombocytopenic purpura,systemic lupus erythematosus,psoria-sis,and looks forward to the prospect of nanobodies in the treatment of autoimmune diseases.
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Interstitial lung disease(ILD)is one of common pulmonary complications of connective tissue disease(CTD),which seriously affects quality of life and significantly increases risk of death of patients with CTD.However,related immune mecha-nism of CTD-ILD is not clear yet.This paper systematically reviews involvement of various cytokines in pathogenesis of CTD-ILD.Pathogenesis of CTD-ILD and correlation between cytokines and clinical indicators were discussed from cell level,animal model and clinical trials,to investigate whether cytokines can be targeted for treatment of CTD-ILD to help guide future research and clinical practice.
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Objective:To explore the correlation between attempted suicide and childhood trauma as well as inflammatory cytokines levels in adolescents with depressive disorder.Methods:From January to December 2021, a total of 164 adolescents with depressive disorder were included and categorized into attempted suicide group ( n=108) and non-attempted suicide group ( n=56) according to whether they accompanied attempted suicide or not. The Chinese version of the suicide module of MINI 5.0, the childhood trauma questionnaire(CTQ), and the center for epidemiological studies depression scale (CES-D) were used to assess suicidal behavior, childhood trauma and depressive symptoms, while the plasma inflammatory cytokines levels were measured by electrochemiluminescence technology. The data were statistically analyzed by SPSS 23.0 software. Logistic regression was applied to analyze the independent influencing factors of attempted suicide and the predictive value of each independent risk factor for attempted suicide was assessed by receiver operating characteristic curve(ROC) curves. Results:The prevalence of attempted suicide was 65.9%(108/164) in adolescents with depressive disorder.The univariate analyses showed that the total CTQ score ((55.19±12.44), (45.50±12.11)) ( t=-4.771, P<0.001), CES-D total score ((40.75±10.26), (32.98±13.39)) ( t=-4.131, P<0.001), IL-6 level(1.87 (1.32, 2.69) pg/mL, 1.56 (1.05, 1.87) pg/mL)( Z=-3.102, P=0.002), IL-10 level (0.53 (0.38, 0.83) pg/mL, 0.47 (0.31, 0.69) pg/mL) ( Z=-2.100, P=0.036) and IL-17A level (2.20 (1.52, 3.65) pg/mL, 1.67 (1.25, 2.97) pg/mL) ( Z=-2.254, P=0.024) in plasma of patients in the attempted suicide group were significantly higher than those in non-attempted suicide group. Logistic stepwise regression analyses showed that CTQ total score, IL-6, IL-10, and IL-17A were risk factors for attempted suicide in adolescents with depressive disorder.The ROC showed that the total score of CTQ combined with IL-6 (AUC=0.782, 95% CI=0.709-0.855, P<0.001), the total score of CTQ combined with IL-10 (AUC=0.765, 95% CI=0.688-0.841, P<0.001), and the total score of CTQ combined with IL-17A (AUC=0.762, 95% CI=0.684-0.840, P<0.001) had a larger area under the curve, indicating better recognition and prediction ability for attempted suicide, with high sensitivity and specificity. Conclusion:The incidence of attempted suicide in adolescents with depressive disorder is high, and which is closely related to childhood trauma and the levels of inflammatory cytokines.
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AIM To explore the mechanism of Yiqi Wenyang Huwei Decoction on airway inflammation improvement of rats with bronchial asthma based on IL-25/NF-κB signaling pathway.METHODS 60 rats were randomly divided into the control group,the model group,the dexamethasone group(0.2 mg/mL),the low-dose,medium-dose and high-dose Yiqi Wenyang Huwei Decoction groups(1,2,4 g/mL),with 10 rats in each group.Intraperitoneal injection of ovalbumin(OVA)and aluminum hydroxide suspension was applied to establish the rat asthma model,followed by 2-week corresponding dosing of the drugs.The rats of each group had their daily diet,mental status,hair growth and respiration observed;their differential count of inflammatory cells in bronchoalveolar lavage fluid(BALF)detected by automatic hematology analyzer;their pathological changes of lung tissue observed by HE staining;their pulmonary IL-25 protein expression detected by immunohistochemistry(IHC);their levels of IL-4,IL-5 and IL-13 in BALF measured by ELISA;their pulmonary expression of IL-25 and TRAF6 mRNA detected by RT-qPCR;and their pulmonary protein expressions of IL-25,TRAF6,IκBα,p-IκBα,NF-κB p65 and p-NF-κB p65 detected by Western blot.RESULTS Compared with the control group,the model group displayed severe damage of the lung tissue and infiltration of a large number of inflammatory cells;increased number of inflammatory cells and levels of IL-4,IL-5 and IL-13 in BALF(P<0.01);increased mRNA expressions of IL-25 and TRAF6,and pulmonary protein expressions of IL-25,TRAF6,p-IκBα/IκBα and p-NF-κB p65/NF-κB p65(P<0.01).Compared with the model group,all of the Yiqi Wenyang Huwei Decoction groups shared improved pulmonary infiltration of inflammatory cells;decreased number of inflammatory cells and levels of IL-4,IL-5 and IL-13 in BALF(P<0.05,P<0.01);and decreased mRNA expressions of IL-25 and TRAF6,and pulmonary protein expressions of IL-25,TRAF6,p-IκBα/IκBα and p-NF-κB p65/NF-κB p65(P<0.01).CONCLUSION Yiqi Wenyang Huwei Decoction can inhibit the airway inflammation in the rat model of bronchial asthma,which may be related to the inhibited activation of IL-25/NF-κB signaling pathway and the reduced expression of inflammatory factors.
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Objective To investigate the anti-inflammatory effects of Bupi Yichang Pills on mice with experimental colitis and its potential mechanism of action.Methods Dextran sulfate sodium(DSS)was used to model the experimental colitis,and low-,medium-and high-doses of Bupi Yichang Pills(1.5,3.0,6.0 g·kg-1·d-1)and Mesalazine(300 mg·kg-1·d-1)were fed at the same time.Mice were observed for general behavior and weighed.Hematoxylin-eosin staining was used to observe the pathological injury of colonic tissues.qPCR and ELISA were used to detect the levels of inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10,IL-35 and TGF-β1),qPCR and Western Blot were used to detect the mRNA and protein levels of glucose transporters and glycolytic kinases.Results Low-,medium-and high-doses of Bupi Yichang Pills significantly down-regulated disease activity index in colitis mice(P＜0.05,P＜0.01).The body mass and colon length were significantly increased,while colon mass,colon mass index and unit colon mass index were significantly reduced(P＜0.05,P＜0.01),and ulcer formation and inflammatory cell infiltration in colonic tissue were significantly improved.In addition,medium-and high-doses of Bupi Yichang Pills significantly down-regulated the mRNA levels and concentrations of pro-inflammatory cytokines including TNF-α,IL-1β and IL-6(P＜0.01),while significantly up-regulated the mRNA levels and concentrations of anti-inflammatory cytokines such as IL-10,IL-35 and TGF-β1(P＜0.01).We further found that high-dose of Bupi Yichang Pills significantly down-regulated the mRNA and protein expressions of glucose transporters(Glut1,Glut2,Glut4)and glycolytic kinases(HK2,Aldolase A,PKM2)in colonic tissue(P＜0.01).Conclusions Bupi Yichang Pills effectively alleviates DSS-induced experimental colitis,and its specific mechanism of action is related to the improvement of glycolytic metabolic pathways and the regulation of inflammatory cytokine expression.
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ObjectiveTo explore the mechanism of Jiawei Wendantang in preventing and treating diabetic atherosclerosis by observing the effect of this prescription on the nuclear factor-κB / NOD-like receptor protein 3（NF-κB/NLRP3） pathway and related inflammatory cytokines in rat model of diabetic atherosclerosis. MethodFifty-four SPF-grade rats were randomized into blank， model， atorvastatin （0.9 mg·kg-1·d-1）， and high-， medium-， low-dose （18.2， 9.1， 4.55 g·kg-1·d-1， respectively） Jiawei Wendantang groups. The rats in other groups except the blank group were modeled for diabetic atherosclerosis by intraperitoneal injection of streptozotocin and feeding with a high-sugar high-fat diet， and those in the blank group were injected with an equal dose of citric acid buffer and fed with a regular diet. The drug administration lasted for 4 weeks， and the blood glucose level in the tail vein was measured every 6 days. After the last administration， the rats were anesthetized for sample collection. Enzyme-linked immunosorbent assay was employed to measure the serum levels of interleukin-18 （IL-18）， C-reactive protein （CRP）， tumor necrosis factor-α （TNF-α）， and intercellular adhesion molecule-1 （ICAM-1）. Western blot was employed to determine the relative protein levels of NF-κB p65， NLRP3， and ICAM-1 in the abdominal aorta. Real-time quantitative polymerase chain reaction was employed to determine the mRNA levels of NLRP3 and interleukin-1β （IL-1β） in the abdominal aorta. The pathological changes in the thoracic aorta were observed by hematoxylin-eosin staining. ResultCompared with the blank group， the model group showed elevated levels of IL-18， CRP， TNF-α， and ICAM-1 in the serum and blood glucose （P<0.05， P<0.01）， up-regulated protein levels of NF-κB p65， NLRP3， and ICAM-1 （P<0.01）， and up-regulated mRNA levels of NLRP3 and IL-1β （P<0.05）. Compared with model group， Jiawei Wendantang lowered the levels of IL-18， CRP， TNF-α， ICAM-1 and blood glucose （P<0.05， P<0.01）， down-regulated the protein levels of NF-κB p65， NLRP3， and ICAM-1 （P<0.01）， and down-regulated the mRNA levels of NLRP3 and IL-1β （P<0.05， P<0.01）. Moreover， Jiawei Wendantang alleviated the pathological injuries in the thoracic aorta. ConclusionJiawei Wendantang may modulate the NF-κB/NLRP3 signaling pathway to reduce the release and adhesion of inflammatory cytokines and regulate the blood glucose level to treat diabetic atherosclerosis.
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Vernal keratoconjunctivitis is an allergic ocular surface disease mostly present with recurrent chronic inflammation, which usually affects children and adolescents and possibly results in corneal complications such as keratoconus. Keratoconus is a corneal disease characterized by a local cone-like corneal deformation. Previous studies have respectively put forward pathological mechanisms of vernal keratoconjunctivitis and keratoconus and the progressive risk factors, among which we find there's mutual mechanisms as well as cytokines' expression, including type I and IV hypersensitivity, inflammatory reaction, enzymatic changes, oxidative stress and mechanical injury. This review aims at summarizing the possible intrinsic mechanisms and cytokines exacerbating vernal keratoconjunctivitis patients corneas to keratoconus, so as to provide reference for the prevention and management in keratoconus caused by vernal keratoconjunctivitis.
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OBJECTIVE To evaluate the impact of sublingual immunotherapy(SLIT)on the balance of Treg/Th17 cells and related cytokines in preschool children aged 3-6 years with allergic rhinitis(AR).METHODS Seventy preschool children aged 3-6 years with AR were divided into the SLIT group and the medication group,and their clinical data were retrospectively analyzed.The medication group received symptomatic treatment alone,while the SLIT group received a combined treatment of SLIT and symptomatic medication,with a 3-year follow-up period.The Treg/Th17 cell balance,serum levels of TGF-β,IL-10,IL-17,IL-21,as well as the total nasal symptom score(TNSS),total medication score(TMS),and visual analog scale(VAS)scores were compared before and after treatment between the two groups.RESULTS After 3 years of treatment,both groups showed significant improvements(P<0.05)in the percentages of CD4+CD25+Foxp3+Treg and CD4+IL-17+Th17 cells among CD4+T cells,percentages of Treg and Th17 cells,serum levels of TGF-β,IL-10,IL-17,IL-21,TNSS,TMS,and VAS scores.Moreover,the SLIT group exhibited significantly better outcomes compared to the medication group(P<0.05).CONCLUSION SLIT can modulate the balance of Treg/Th17 cells and improve serum levels of TGF-β,IL-10,IL-17,and IL-21.
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Objective To analyze the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of Mycoplasma pneumoniae pneumonia(MPP)in children.Methods A total of 120 children with MPP who were admitted to the hospital from January 2020 to January 2023 were selected as the observation group,80 children with pulmonary tuberculosis(TB)were selected as the control group.During the same pe-riod,120 healthy children who were examined at the hospital check-up center were selected as the health group.The clinical data from each group were retrospective analyzed,and fasting venous blood from subjects was collected.The levels of T-lymphocyte subsets CD3+,CD4+and CD8+were detected by flow cytometry in each group,and the CD4+/CD8+was calculated.The levels of interferon(IFN)-y,interleukin-8(IL-8),inter-leukin-10(IL-10)and interleukin-13(IL-13)were measured by enzyme-linked immunosorbent assay,and the levels of peripheral blood T-lymphocyte subsets and cytokines in each group were compared.The receiver op-erating characteristic(ROC)curve was used to assess the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of MPP in children.Results The levels of CD3+,CD4+,CD4+/CD8+,IL-13,and IL-10 in the observation group and control group were significantly lower than those in the health group(P＜0.05),while CD8+,IL-8,and IFN-γ levels were significantly higher than those in the health group(P＜0.05).Compared with the control group,the observation group had higher levels of CD3+,CD8+,IL-8,IFN-γand lower levels of CD4+,CD4+/CD8+(P＜0.05),and there were no statistically significant differences in IL-8 and IL-13 levels between the two groups(P＞0.05).The ROC curve results showed that the area under the curve(AUC)or CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13,and IL-10 for the diagnosis of MPP in children were 0.751,0.687,0.784,0.864,0.798,0.672,0.650,and 0.811,and AUC of the combined detection was 0.924.Conclusion Children with TB and MPP have significantly decreased immune function in the early stages of the disease,and abnormal expression of peripheral blood T-lymphocyte subsets and cytokines.Com-pared with TB children,MPP children have lower levels of CD4+,CD4+/CD8+,and higher levels of CD3+,CD8+,IL-10 and IFN-γ,and the T-lymphocyte subsets and cytokine levels are closely related to the changes in the patients'condition.The combined detection of CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13 and IL-10 provides a theoretical basis for identifying and diagnosing early MPP in children.
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Objective To explore the application value of lymphocyte subsets combined with various cytokines in the disease progression of elderly patients with corona virus disease 2019(COVID-19).Methods From December 2022 to January 2023,146 elderly patients with COVID-19 diagnosed in the emergency ward of the Eighth Medical Center of PLA General Hospital were selected and divided into two groups according to the prognosis:127 cases in the COVID-19 survival group,19 cases in the COVID-19 death group.In addition,51 osteoporosis patients in geriatric medicine department were collected as control group.The proportion and absolute count of lymphocyte subsets(including T,B and NK cells),and 12 cytokines in plasma(including IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,TNF-α and IFN-γ)were compared between the control group and COVID-19 group,survival group and death group.The receiver operating characteristic(ROC)curve was used to evaluate its prognostic value in elderly patients with COVID-19 infection.Results Compared with the control group:① The proportion of NK cells in COVID-19 group was decreased,while the proportion of B cells was increased,and the differences were statistically significant(Z=-3.386,-4.140,all P<0.01).There was no significant difference in the proportion of T,CD8+T and CD4+T cells,and the differences were not statistically significant(Z=-1.244,-1.770,-0.951,all P>0.05).② The absolute numbers of T,CD8+T,CD4+T,NK and B cells in COVID-19 group were decreased,and the differences were statistically significant(Z=-9.418～-6.539,all P<0.01).③ The concentrations of IL-2,IL-6,IL-1β,IFN-γ,IL-8,IL-17,IL-12P70 and IL-10 in COVID-19 group were all increased,and the differences were statistically significant(Z=-8.851～-1.986,all P<0.05).There was no significant difference in the concentrations of IL-5,IFN-α,TNF-α and IL-4,and the differences were not statistically significant(Z=-0.460～-0.217,all P>0.05).Compared with the survival group:① There was no significant difference in the proportion of T,CD8+T,CD4+T,NK and B cells in the death group(Z=-1.873～-0.422,all P>0.05).② The absolute numbers of T,CD8+T and CD4+T cells in the death group were all decreased,and the differences were statistically significant(Z=-2.667,-2.287,-2.556,all P<0.05),while there was no significant difference in absolute numbers of NK and B cellsm and the differences were not statistically significant(Z=-1.934,-0.532,all P>0.05).③ The concentrations of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in the death group were all increased,and the differences were not statistically significant(Z=-4.211～-2.655,all P<0.05),and there was no significant difference in the concentrations of IL-5,IFN-α,IL-2,IL-1β,IL-12p70,TNF-α and IL-4 the differences were not statistically significant(Z=-1.329～-0.279,all P>0.05).ROC curve analysis for the prognostic value of lymphocyte subsets combined with cytokines in elderly patients with COVID-19 showed that:the areas of total T cells,B cells and NK cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.94,0.80 and 0.93,respectively.The areas of CD4+T cells and CD8+T cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.93 and 0.90,respectively.The areas of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in cytokines under the ROC curve for predicting the prognosis of COVID-19 infection were 0.91,0.71,0.87,0.74 and 0.90,respectively.However,the area of combined lymphocyte subsets and cytokines under ROC curve for predicting the prognosis of COVID-19 infection reached 0.99.Conclusion The immune status of elderly patients with COVID-19 was generally low.Evaluation of immune status has important clinical guidance significance in disease diagnosis,disease observation and prognosis.
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Objective To analyze the predictive value of serum levels of procalcitonin(PCT)and cytokines on the prognosis of patients with COVID-19 at admission.Methods From November 2022 to February 2023,patients diagnosed with COVID-19 who were admitted to Beijing Chest Hospital were enrolled.Chemiluminescence was used to detect serum PCT levels,and flow microsphere array was used to detect serum cytokines IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17A,IL-17F,IL-22,TNF-α,TNF-β,IFN-γ level.ICU admission,mechanical ventilation and in-hospital death were defined as poor prognosis.After excluding patients with bacterial infection,the relationship between serum PCT and cytokine levels at admission and the prognosis of COVID-19 patients was analyzed.After excluding patients with bacterial infection,the relationship between serum PCT and cytokine levels at admission and the prognosis of COVID-19 patients was analyzed.Results A total of 176 patients with complete data were included,including 134 in the PCT-normal group and 42 in the PCT-elevated group,with a median age of 71.50 years and 71.59%males.Patients in the PCT elevated-group had significantly higher rates of ICU admission(38.41%vs.13.11%,P<0.05),mechanical ventilation(76.92%vs.24.59%,P<0.001)and in-hospital mortality(38.46%vs.6.56%,P<0.001)were significantly higher than those in the PCT-normal group.Serum levels of cytokines IL-6(7.40 pg/mL vs.4.78 pg/mL,P = 0.033 4)and IL-8(10.97 pg/mL vs.5.92 pg/mL,P<0.001)were significantly higher in patients with poor prognosis than in those with good prognosis.The area under the curve for PCT,IL-6,and IL-8 to predict poor prognosis in COVID-19 patients was 0.687,0.660,and 0.746,respectively;sensitivity was 52.78%,55.17%,and 72.41%,respectively;and specificity was 81.58%,74.19%,and 74.19%,respectively,as calculated from the ROC curve.When PCT,IL-6 and IL-8 jointly predict the prognosis of COVID-19 patients,the area under the curve is 0.764,the sensitivity is 70.00%,and the specificity is 80.00%.Conclusion Serum PCT and cytokines IL-6 and IL-8 could be used as predictive markers for poor prognosis in patients with COVID-19.
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ObjectiveTo investigate the therapeutic effects and difference in the effects of Arisaematis Rhizoma （AR） before and after processing （i.e.， Arisaematis Rhizoma Preparatum， ARP） with Zingiberis Rhizoma Recens-Alumen on allergic asthma in rats and to provide a basis for the theory of processing improving the efficacy. MethodA rat model of allergic asthma was established in 70 SD rats by intraperitoneal injection of ovalbumin （OVA）-aluminum hydroxide. The rats were administrated with the aqueous extracts of AR （1.2， 0.3 g∙kg-1） and ARP （1.2， 0.3 g∙kg-1） aqueous extracts by gavage， and montelukast sodium （0.001 g∙kg-1） was used as the positive drug. The T helper cell type 1/type 2 （Th1/Th2） ratio in the serum and bronchoalveolar lavage fluid （BALF） and percentages of inflammatory cells in BALF were determined. Polymerase chain reaction （PCR） was employed to determine the mRNA level of mucin 5AC （MUC5AC） in the lung tissue. The pathological changes in the lung tissue were observed by hematoxylin-eosin （HE） staining and PAS staining. Immunohistochemical assay was employed to measure the expression of c-Jun amino-terminal kinase （JNK）， extracellular signal regulated protein kinase （ERK）， and p38 mitogen-activated protein kinase （p38 MAPK） in rat lung tissue. Western blot was employed to determine the protein levels of ERK， p-ERK， JNK， p-JNK， p38， p-p38 in the lung tissue. The effects of AR and ARP were compared based on overall desirability. ResultCompared with the blank group， the levels of interleukin-12 （IL-12） and γ interferon （IFN-γ） in serum and BALF of rats in the model group were significantly lower （P<0.05， P<0.01）， and the levels of interleukin-4 （IL-4）， interleukin-5 （IL-5） and interleukin-13 （IL-13） were significantly higher （P<0.05， P<0.01）. Compared with the model group， the serum and BALF contents of IL-12 and IFN-γ in rats in the montelukast sodium group， high-dose AR group and high-dose ARP group were significantly higher （P<0.05， P<0.01）， and the contents of IL-4， IL-5 and IL-13 were significantly lower （P<0.05， P<0.01）， and the serum contents of IFN-γ in rats in the low-dose AR group and low-dose ARP group were in BALF was significantly higher （P<0.05） and IL-4 and IL-13 were significantly lower （P<0.05， P<0.01）， the percentages of macrophages， lymphocytes， neutrophils， and eosinophils were reduced in BALF， and the expression of JNK/ERK/p38 MAPK signaling pathway and MUC5AC protein was inhibited in lung tissues. Overall assessment of the normalized analysis revealed that the ARP group was slightly more potent than the AR group after administration of the same dose. ConclusionAR and ARP can effectively treat allergic asthma by inhibiting JNK/ERK/p38 MAPK signaling pathway， and the effect is better after concoction， which can provide data support for its "concoction efficiency".
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ObjectiveTo evaluate the clinical efficacy of modified Banxia Xiexintang combined with vedolizumab （VDZ） in the treatment of active moderate to severe Crohn's disease （CD） with the syndrome of cold and heat in complexity and the effect of the therapy on intestinal flora. MethodEighty patients were randomized based on the random number table method into a control group （40 cases） and an observation group （40 cases）. The control group was treated with VDZ， and the observation group was treated with modified Banxia Xiexintang （1 bag per day） combined with VDZ. The treatment in both groups lasted for 14 weeks and the follow-up lasted until the 52th weeks. The CD activity index （CDAI）， CD simplified endoscopic score （SES-CD）， inflammatory bowel disease questionnaire （IBDQ） score， and syndrome score of cold and heat in complexity were assessed before treatment， after treatment， and at the end of follow-up. The levels of hemoglobin （HGB）， hematocrit （HCT）， albumin （ALB）， C-reactive protein （CRP）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-17 （IL-17）， and fecal calprotectin （FC） were measured before and after treatment. Intestinal flora was examined before and after treatment. The safety of the therapy was evaluated. ResultCompared with those before treatment， the scores of CDAI， SES-CD， and the syndrome of cold and heat in complexity decreased （P<0.05） and the IBDQ score increased after treatment （P<0.05）. Compared with those before treatment， the scores of CDAI， SES-CD， and the syndrome of cold and heat in complexity increased （P<0.05） and the IBDQ score decreased （P<0.05） at the end of follow-up. After treatment and at the end of follow-up， the observation group had lower scores of CDAI， SES-CD， and syndrome of cold and heat （P<0.05） and higher IBDQ score （P<0.05） than the control group. Moreover， the observation group had higher clinical remission rate（χ2=4.381，3.962） and response rate（χ2=5.541，4.306） and lower non-response rate（χ2=6.646，4.306） than the control group at the two time points （P<0.05）. The endoscopic remission rate（χ2=4.072，3.985） and response rate（χ2=4.528，5.161） in the observation group were higher than those in the control group （P<0.05）. After treatment， the HGB， HCT， and ALB levels in both groups elevated， and the observation group had higher levels than the control group （P<0.05）. The treatment in both groups lowered the levels of CRP， IL-6， TNF-α， IL-17， and FC （P<0.05）， and the observation group had lower levels of CRP， IL-6， TNF-α， IL-17， and FC than the control group after treatment （P<0.05）. The relative abundance of Bifidobacterium， Lactobacillus， and Prevotella increased （P<0.05）， while that of Proteus， Klebsiella， and Enterococcus decreased （P<0.05） in the two groups after treatment. Moreover， the changes in the relative abundance of these bacteria in the observation group were more obvious than those in the control group （P<0.05）. No adverse reactions related to the modified Modified Banxia Xiexintang were observed during the study period. ConclusionModified Banxia Xiexintang combined with VDZ can play a synergistic role and has good short-term and long-term efficacy. This therapy can improve the nutritional status， regulate intestinal flora， and reduce inflammatory injury in the treatment of moderate to severe active CD patients with the syndrome of cold and heat in complexity without causing severe adverse reactions.