Study on mechanism and Huanglong Mixture in the treatment of children with cough variant asthma and experimental verification based on network pharmacologic analysis / 国际中医中药杂志

Objective:To study the the mechanism of action of Huanglong Mixture in the treatment of cough variant asthma (CVA) in children based on the IL-4/signal transduction and activator of transcription 6 (STAT6) signaling pathway using network pharmacology methods, molecular docking techniques, and in vitro cell experiments.Methods:The components and targets of various TCM components in Huanglong Mixture were searched in TCMSP database, HERB database and literature, and the disease targets of CVA were found in Gene Cards database, OMIM database, DrugBank database and PharmGkb database. The STRING database was used to construct the protein-protein interaction network, and Cytoscape 3.9.1 was used for topology analysis to screen out the core targets. The disease-drug-component-target network was constructed to screen out the core components. The KEGG enrichment analysis and GO enrichment analysis of the intersection targets were performed using Metascape software. PDB protein database, PubChem, Autodock and R language were used for molecular docking verification of core targets and core drug components. Finally, rat primary airway smooth muscle cells were cultured, modeled with interleukin-4 (IL-4), and p-STAT6 expression in the cytoplasm and nucleus was detected by Western blot.Results:A total of 122 effective components were obtained, including quercetin, kaempferol, luteolin and so on. The core targets included JUN, ESR1, TP53, MYC, HIF1, etc. GO enrichment analysis involved biological processes such as response to external stimuli, response to oxygen levels, positive regulation of protein phosphorylation, and regulation of cellular stress response. KEGG enrichment analysis showed that the main pathways of Huanglong Mixture in treating CVA included advanced glycation end product-glycation end product receptor (AGE-RAGE) signaling pathway, phosphatidylinositol-3-kinase-protein kinase B (PI3K-Akt) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Janus kinase/signal transduction activation factor (JAK-STAT) signaling pathway. Molecular docking found that the core targets and core drug components had good combination. Cell experiments also confirmed that Huanglong Mixture could inhibit p-STAT6 entering the nucleus.Conclusions:The effective components and targets of Huanglong Mixture in the treatment of CVA are successfully predicted. The mechanism of Huanglong Mixture in the treatment of children with CVA may be related to the inhibition of IL-4/STAT6 signaling pathway.

Correlation of serum RAGE and HMGB1 expression with the occurrence of acute respiratory distress syndrome and IFN-γ/IL-4 ratio in patients with severe pneumonia / 实用医学杂志

Objective To explore the correlation between the expression level of serum Receptor for Advanced Glycation End-Product(RAGE)and High-Mobility Group Protein B1(HMGB1)expression with the occurrence of acute respiratory distress syndrome(ARDS)and interferon-γ/interleukin-4(IFN-γ/IL-4)ratio in patients with severe pneumonia(SP).Methods A prospective investigation was carried out on one hundred children with SP admitted to our hospital from March 2020 to February 2022,and the participants were classified into ARDS group(n = 56)and control group(n = 44)based on the occurrence of secondary ARDS.General informations werec-ollected.The expression of RAGE,HMGB1,IFN-γ and IL-4 in peripheral blood was measured using Enzyme-Linked Immunosorbent Assay(ELISA).Then multivariate Logistic regression analysis was conducted to screen the influencing factors of secondary ARDS in SP children,and the correlation with IFN-γ/IL-4 ratio was verified by pearson correla-tion analysis,moreover,receiver operating characteristic(ROC)curve was plotted to evaluate the value of RAGE and HMGB1 expression in predicting the occurrence of ARDS in SP children.Results There were no statistical difference in gender,age,body temperature and onset season between the two SP groups.The ARDS group had more types of pathogenic bacteria,larger ratio of the partial pressure of oxygen in arterial blood to the inspired oxygen fraction(PaO2/FiO2),higher Acute Physiological Score(APS),and up-regulated expression of RAGE,HMGB1,IFN-γ and IL-4,as well as larger IFN-γ/IL-4 ratio than those of control group,with statistical difference(all P<0.05).Multivariate Logistic regression analysis revealed that pathogen type,PaO2/FiO2 ratio,RAGE,HMGB1,IFN-γ,IL-4 and IFN-γ/IL-4 were the influencing factors for the occurrence of ARDS in children with SP.Pearson correlation test denoted that the serum RAGE and HMGB1 expression levels of SP children were positively correlated with IFN-γ,IL-4 and IFN-γ/IL-4 ratio(P<0.05).ROC curve found that the AUC of serum RAGE and HMGB1 in predicting the occurrence of ARDS in SP children was 0.707 and 0.750,with a sensitivity of 73.2%and 64.3%,and a specificity of 68.2%and 77.3%.The combined test of RAGE and HMGB1 in predicting the occurrence of ARDS in SP children reached an AUC of 0.848,providing a sensitivity and specificity of 80.4%and 81.8%respectively.Conclusions Serum RAGE and HMGB1 expression levels are elevated in SP children with ARDS,and the two are positively correlated with IFN-γ/IL-4 ratio.Therefore,monitoring serum RAGE and HMGB1 expression in children with ARDS secondary to SP has predictive value for the risk of ARDS in SP children.

Evaluation of IL-4, MIP-1α, and MMP-9 gene expression levels in peri-implant tissues in peri-implantitis

Abstract This case-control study evaluated the gene expression levels of interleukin (IL)-4, macrophage inflammatory protein type 1 alpha (MIP-1α), and metalloproteinase (MMP)-9, factors involved in the formation of giant cells in healthy peri-implant tissue and peri-implantitis. Thirty-five subjects (15 healthy and 20 with peri-implantitis), who met the inclusion and exclusion criteria, were included in this study. The peri-implant tissue biopsies were subjected to total RNA extraction, DNAse treatment, and cDNA synthesis. Subsequently, the reaction of real-time PCR was performed to evaluate the gene expression levels of IL-4, MIP-1α, and MMP-9 concerning the reference gene. IL-4 gene expression showed higher (18-fold) values in the Peri-Implantitis Group of Patients when compared with the Healthy (Control) Group (p<0.0001). Although MIP- 1α and MMP-9 gene expression levels were higher in diseased implants, they showed no significant differences (p=0.06 and p=0.2337), respectively. Within the limitations of this study, the results showed that in tissues affected by peri-implantitis, only levels of Il-4 were increased when compared with tissues in the control group.

Resumo Este estudo caso-controle teve como objetivo avaliar a expressão gênica dos níveis de interleucina (IL)-4, proteína inflamatória de macrófagos tipo alfa 1 (MIP-1α) e metalopreoteinase (MMP)-9, todos fatores envolvidos na formação de células gigantes em tecidos peri-implantares saudáveis e com peri-implantite. Trinta e cinco indivíduos (15 saudáveis e 20 com peri-implantite) foram incluídos nesse estudo seguindo os critérios de inclusão e exclusão. Os tecidos peri-implantares foram submetidos a extração do RNA total, tratamento de DNAse e síntese de cDNA. Subsequentemente, a reação de PCR em tempo real foi realizada para avaliar os níveis da expressão de IL-4, MIP-1α, e MMP-9 em relação ao gene de referência. O nível de expressão de IL-4 foi estatisticamwente maior (18 vezes) nos tecidos de pacientes com peri-implantite quando comparados aos pacientes saudáveis (grupo controle) (p<0,0001). Embora os níveis de expressão de MIP- 1α e MMP-9 apresentassem maiores valores nos implantes doentes, esses níveis não foram estatisticamente significantes (p=0.06 and p=0.2337) respectivamente. Dentro das limitações desse estudo, os resultados mostraram que nos tecidos afetados pela peri-implantite, apenas os nívies de IL-4 estavam aumentados quando comparados ao grupo controle.

Epidemiological characteristics of allergic rhinitis and its correlation with serum IFN-γ and IL-4 levels / 公共卫生与预防医学

Objective To analyze the epidemiological characteristics of AR and its correlation with serum IFN-γ and IL-4 levels in Bazhong City, and to provide theoretical basis for clinical prevention and treatment of AR. Methods Adopt the method of multistage stratified random survey from January 2019 to January 2020 bazhong 3 armour hospital otolaryngology seeing a doctor , with face to face questionnaire survey form the people generally, AR number of statistics, the main clinical symptoms and related symptoms, AR merger disease situation, according to the illness severity was divided into mild and moderately severe group, Five mL of elbow venous blood was extracted from AR patients, and 22 kinds of allergens were determined by using the allergen-specific IgE antibody detection kit (western blot). Serum IFN-γ and IL-4 levels were determined by enzyme-linked immunosorbent assay. Pearson correlation analysis was performed on serum IFN-γ and IL-4 levels and severity of allergic rhinitis. Results Among 1 243 patients who completed the questionnaire, the prevalence of AR was 275 (22.12%). There was significant difference in the prevalence of AR among different age groups (χ2=6.809 , P2=7.914 , P<0.0) and the prevalence of AR in workers was the highest (26.48%). Among 275 AR patients in Bazhong City, 153 cases (55.64%) had seasonal allergic rhinitis and 122 cases (44.36%) had perennial allergic rhinitis. The main clinical symptoms of AR patients were sneezing in 234 cases (85.09%) and nasal obstruction in 197 cases (71.64%). AR combined with bronchial asthma in 59 cases (21.45%), conjunctivitis in 28 cases (10.18%); the peak of AR incidence was mainly in July. Among 22 allergens, dust mite was the most common allergen in 139 cases (50.55%), followed by penicillin in 56 cases (20.36%). The main inducing factors were pollen 112 cases (40.73%) and cold 98 cases (35.64%). According to the severity of the disease, they were divided into mild group (n=178) and moderate to severe group (n=97). The serum IL-4 level in moderate and severe groups was significantly higher than that in mild group (P<0.05). The serum IFN-γ level in moderate and severe groups was significantly lower than that in mild group (P<0.05). Pearson correlation analysis showed that il-4 level was positively correlated with disease severity (r=0.492, P<0.05). IFN-γ was negatively correlated with the severity of the disease (r=-0.459, P<0.05). Conclusion The prevalence of AR is high in Bazhong city, and the main clinical symptom is sneezing. Among the complications, bronchial asthma is the most common. Colds and weather changes are the main factors causing AR.

Changes in serum IL-1β, IFN-γ and IL-4 levels in children with sepsis and their values in differential diagnosis of Gram-positive/Gram-negative bacterial infection / 中华微生物学和免疫学杂志

Objective:To analyze the changes in serum IL-1β, IFN-γ and IL-4 levels in children with sepsis and analyze their values in the differential diagnosis of Gram-positive (G + )/Gram-negative (G -) bacterial infection. Methods:Clinical data of 195 children with sepsis admitted to Xuzhou Central Hospital from February 2020 to February 2023 were retrospectively analyzed. They were all confirmed to have G + /G - bacterial infection based on pathogen culturing and identification and enrolled as disease group. Another 180 healthy children taking physical examination in the same hospital during the same period were recruited as control group. The distribution of G + /G - bacterial infection in the disease group was analyzed after pathogen culturing. ELISA was used to detect serum IL-1β, IFN-γ and IL-4 levels in both groups. Differences in the levels of serum IL-1β, IFN-γ and IL-4 were compared between the disease group and the control group as well as between the patients with G + /G - bacterial infection. Receiver operating characteristic (ROC) curve was drawn to evaluate the values of serum IL-1β, IFN-γ and IL-4 levels used alone or in combination in the differential diagnosis of G + /G - bacterial infection. Results:A total of 211 strains were isolated from 195 blood culture samples in the disease group, including 181 strains (85.78%) of G + bacteria and 30 strains (14.22%) of G - bacteria. Coagulase-negative Staphylococcus and Salmonella were the predominant bacteria causing G + /G - bacterial infection, respectively. The levels of serum IL-1β and IFN-γ in the disease group were higher than those in the control group ( P<0.05), while the levels of serum IL-4 were similar in the two groups ( P>0.05). The levels of serum IL-1β, IFN-γ and IL-4 in the patients with G - bacterial infection were higher than those in the patients with G + bacterial infection ( P<0.05). The sensitivity and the area under the ROC curve of serum IL-1β, IFN-γ and IL-4 levels used in combination in the differential diagnosis of G + /G - bacterial infection were respectively 91.62% and 0.960, which were higher than those of each parameter used alone ( P<0.01, P<0.05). However, there was no significant difference in the specificity ( P>0.05). Conclusions:The levels of serum IL-1β and IFN-γ increased in children with sepsis, while the serum IL-4 level was basically normal in them. Children with G - bacterial infection tended to have higher serum IL-1β, IFN-γ and IL-4 levels. The combination use of all three parameters showed higher value in the differential diagnosis of G + /G - bacterial infection.

Effect and mechanism of Asperosaponin Ⅵ on polarization of M1/M2 macrophages / 中国免疫学杂志

Objective:To study the regulation and mechanism of Asperosaponin Ⅵ on polarization of M1/M2 macrophages.Methods:MTT assay was used to detect the effects of Asperosaponin Ⅵ on RAW264.7 cell viability.The levels of TNF-α and IL-6 in supernatant of RAW264.7 cells induced by lipopolysaccharide(LPS)were determined by ELISA.The content of nitric oxide(NO)in supernatant of RAW264.7 cells induced by LPS was determined by Griess method.The gene expression levels of TNF-α,IL-6,argi-nase-1(Arg-1),heme oxygenase-1(HO-1)and suppressor of cytokine signaling protein-2(SOCS2)were detected by fluorescence quantitative PCR.Western blot was used to detect the expression levels of iNOS and p-p65 protein.Results:In LPS induced RAW264.7 cells,Asperosaponin Ⅵ inhibited protein or gene expression levels of TNF-α,IL-6,iNOS and p-p65,and increased HO-1 gene expression.Asperosaponin Ⅵ inhibited NO secretion in RAW264.7 cells induced by LPS.Asperosaponin Ⅵ increased the gene expression levels of M2 macrophage markers Arg1 and SOCS2 induced by IL-4.Conclusion:Asperosaponin Ⅵ inhibited RAW264.7 macrophage polarization to M1 type and promote it polarization to M2 type,which can play its anti-inflammatory and immunomodulato-ry role by regulating M1/M2 macrophage polarization.

Acupuncture combined with western medication for ocular myasthenia gravis: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical efficacy between acupuncture combined with western medication and simple western medication for ocular myasthenia gravis (OMG), and to explore its possible mechanism.@*METHODS@#A total of 60 patients of ocular myasthenia gravis were randomized into an acupuncture combined with western medication group (30 cases, 1 case dropped off) and a western medication group (30 cases, 2 cases dropped off). Oral pyridostigmine bromide tablet and prednisone acetate tablet were given in the western medication group. On the basis of the treatment in the western medication group, Tongdu Tiaoqi acupuncture (acupuncture for unblocking the governor vessel and regulating qi ) was applied at Baihui (GV 20), Fengfu (GV 16), Hegu (LI 4), Zusanli (ST 36), etc. in the acupuncture combined with western medication group, once a day, 6 days a week. The treatment was given 8 weeks in both groups. Before and after treatment, the OMG clinical absolute score was observed, electrophysiological indexes of orbicularis oculi (value of mean jitter, percentage of jitter >55 μs and percentage of blocks) were measured by single-fiber electromyography (SFEMG), serum levels of acetylcholine receptor antibody (AChR-Ab), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were detected by ELISA method.@*RESULTS@#After treatment, the OMG clinical absolute scores, values of mean jitter, percentages of jitter >55 μs, percentages of blocks and serum levels of AChR-Ab, IFN-γ and IL-4 were decreased compared before treatment in both groups (P<0.05), and those in the acupuncture combined with western medication group were lower than the western medication group (P<0.05).@*CONCLUSION@#Acupuncture combined with western medication can effectively improve ptosis, palpebra superior fatigability, eye movement disorder and neuromuscular junction dysfunction in patients with ocular myasthenia gravis, the therapeutic effect is superior to simple western medication. Its mechanism may be related to down-regulating serum levels of AChR-Ab, IFN-γ and IL-4 and promoting the recovery of orbicularis oculi function.

Effect of Maxing Shigantang on Airway Injury and EGFR Expression in Asthma Model Rats / 中国实验方剂学杂志

ObjectiveTo explore the optimal formula of Maxing Shigantang in regulating epidermal growth factor receptor（EGFR）expression and alleviating airway injury in asthmatic rats and to reveal the underlying mechanism. MethodSD male rats were randomly divided into normal group, model group, dexamethasone group (5×10-4 g·kg-1) and Maxing Shigantang 1∶0.5, 1∶1, 1∶2 groups (group A, B, C, 10 g·kg-1), with 8 rats in each group. The other groups except the normal group received nebulization of 2% acetylcholine chloride and 0.4% histamine phosphate for the modeling of asthma. One hour before modeling， the normal group and the model group were given the same amount of normal saline， and the other groups were given the same amount of corresponding drugs， once a day for 7 days. On the 7th day， the model was established and the incubation period of asthma was recorded. The rats were then immediately anesthetized， and arterial blood and tracheal tissue were collected. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the levels of interleukin-2 （IL-2）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in serum. Pathological sections were prepared for the observation of the pathological changes of tracheal tissues and the ultrastructure of epithelial cells in each group. Terminal-deoxynucleotidyl transferase-mediated nick-end labeling （TUNEL） was adopted to detect epithelial cell apoptosis， and in situ hybridization and Western blot were employed to determine the mRNA and protein levels of epidermal growth factor receptor （EGFR）， respectively. ResultCompared with the model group， groups A， B and C prolonged the incubation period of asthma （P<0.05，P<0.01）. Compared with the control group， the model group showed declined IL-2 level （P<0.01）， risen IL-4 and TNF-α levels （P<0.05，P<0.01）， increased airway pathology score， collagen volume fraction， and airway epithelial cell apoptosis index （P<0.01）， and up-regulated mRNA and protein levels of EGFR in trachea tissue （P<0.01）. Compared with the model group， group A showed increased IL-2 level （P<0.05） and declined IL-4 （P<0.05，P<0.01） level， and group B showed declined IL-4 level （P<0.05）. The level of TNF-α in groups A， B， and C declined compared with that in the model group （P<0.01）. Maxing Shigantang repaired the tracheal tissue to different degrees （P<0.05）. Among the three groups， group A inhibited tracheal fibrosis （P<0.05） and had the most significant effect of repairing the ultrastructural changes of airway epithelial cells. Groups A， B and C all inhibited the apoptosis of airway epithelial cells （P<0.05）. All the three groups inhibited the up-regulation of EGFR mRNA level （P<0.05，P<0.01）， and groups B and C inhibited the up-regulation of EGFR protein level （P<0.05，P<0.01）. ConclusionMaxing Shigantang can inhibit the abnormal changes of airway epithelial structure， alleviate airway injury， and can down-regulate the expression of EGFR in the tracheal tissue of asthma model rats. In this study， the optimal compatibility of Maxing Shigantang to repair airway epithelial injury in asthmatic rats was group A， with the Ephedrae Herba-Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma-Gypsum Fibrosum ratio of 1∶0.5∶4∶1.

Inhibitory effect of targeted fusion peptide IL-4Rα-lytic on primary effusion lymphoma / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨靶向融合肽IL-4Rα-lytic对卡波西肉瘤相关疱疹病毒（KSHV）阳性原发性渗出性淋巴瘤（PEL）细胞的体内外杀伤作用及其安全性。方法：应用MTT法检测IL-4Rα-lytic对KSHV阳性PEL细胞BCBL-1和BCP-1的杀伤能力。通过FCM检测IL-4Rα-lytic诱导KSHV阳性PEL细胞凋亡的情况。建立BCBL-1细胞小鼠移植瘤模型，连续3周（3次/周）腹腔注射IL-4Rα-lytic后，通过活体生物发光成像技术评估IL-4Rα-lytic对小鼠体内BCBL-1细胞移植瘤的抑制效果，并通过H-E染色和全血分析法检测其毒副作用。结果：靶向融合肽IL-4Rα-lytic在体外对两种KSHV阳性PEL细胞BCBL-1和BCP-1均有选择性杀伤作用（均P<0.01），并且可以在短时间内发挥杀伤作用（均P<0.01）。靶向融合肽IL-4Rα-lytic可诱导KSHV阳性PEL细胞BCBL-1和BCP-1凋亡（均P<0.05）。靶向融合肽IL-4Rα-lytic显著抑制BCBL-1细胞小鼠移植瘤的生长，与对照组比较差异具有统计学意义（P<0.05），并且无明显的器官毒性（均P>0.05），同时不会造成体质量异常（P>0.05）。结论：靶向融合肽IL-4Rα-lytic在体内外均显著抑制KSHV阳性PEL细胞的生长，且无明显毒副作用，有望为PEL的治疗提供一种新的治疗方案。

Effect mechanism of Ma Xing Gan Shi Decoction against influenza virus from JAK2/STAT3 signaling pathway by combining network pharmacology / 中国药理学通报

Aim To explore the effect of JAK2/STAT3 signaling pathway on lung tissue injury induced by influenza A virus in combination with network pharmacology and to further explore the intervention effect of Ma Xing Gan Shi Decoction.Methods Network pharmacological method was used to screen the signal pathway enriched by Ma Xing Gan Shi Decoction on the potential target of influenza virus.BLAB/c mice were intranasally infected with influenza A virus.The mice were divided into normal control group, model control group, oseltamivir group, antiviral granule group and Ma Xing Shigan decoction group.The animals were treated with corresponding drugs for 3 and 7 days.Body weight and lung index were detected by HE for observation of the pathological changes of lung tissues.Real-time PCR(RT-PCR)was used to detect the mRNA expression levels of JAK2, STAT3, IL-1β and IL-4 in lung tissues.Western blot and ELISA were used to detect the protein expression levels of JAK2, STAT3, IL-1β and IL-4 in lung tissues.AutoDock Vina software was used to conduct molecular docking between STAT3 and target compounds.Results The main active components of Ma Xing Gan Shi Decoction had 110 intersection targets with influenza virus and were enriched in 170 signaling pathways.Ma Xing Shigan decoction could up-regulate the body weight of mice infected with influenza A virus, improve the pathological injury of lung tissues, down-regulate the lung index and the expression levels of JAK2, STAT3, IL-1β mRNA and protein in lung tissues, and up-regulate the expression levels of IL-4 mRNA and protein.STAT3 had better binding activity with glycyrrhiza chalcone A, an active compound in Ma Xing Gan Shi Decoction.Conclusions Ma Xing Gan Shi Decoction, as an effective compound prescription of traditional Chinese medicine against influenza virus, can effectively reduce pulmonary inflammation and regulate the balance of cytokines.The possible mechanism is to alleviate the lung injury caused by influenza A virus infection in mice by inhibiting the activation of JAK2/STAT3 signal pathway.

Expresión génica de mediadores moleculares para potencial terapia celular en el síndrome gastrointestinal inducido por radiación / Genetic expression of molecular mediators for potential cell therapy in radiation-induced gastrointestinal syndrome

Antecedentes: Las células madres intestinales generan las distintas estirpes celulares a dicho nivel. Estas se regulan por interacciones entre el epitelio y las células del nicho celular anexo. Estas se pueden ver dañadas en tratamientos con radiación, generando el síndrome gastrointestinal inducido por radiación. Se ha visto que células madre mesenquimales (MSC) y macrófagos de médula ósea (BMM) tienen propiedades de regeneración tisular. Objetivos: Evaluar la expresión génica de IL-4, Wnt6, VEGF y bFGF, a partir de cultivos celulares primarios independientes de MSC derivadas de tejido adiposo y BMM de ratones C57BL/6, por medio de PCR en tiempo real (qRT-PCR). Diseño experimental: A partir de un análisis in silico, se confeccionaron primers para evaluar la expresión génica de las moléculas propuestas, en los cultivos primarios por medio de qRT-PCR y electroforesis. Resultados y proyecciones: IL-4 y Wnt6 no son expresadas en las muestras de BMM y MSC. VEGF y bFGF son expresadas por diferentes células, dando expresión diferenciada. A futuro, se deben evaluar las mismas estirpes celulares en un ambiente inflamatorio y su efecto en la expresión génica, en especial VEGF y bFGF. Limitaciones: El número de moléculas en estudio es limitado y la expresión se evalúo solo a nivel genético.

Background: Intestinal stem cell generates diferents cellular types in their niche. They're regulated by interactions between epithelium and niche's cells, and can be damaged by medical radiation treatments causing radiation-induced gastrointestinal syndrome. It has seen that mesenchymal stem cells (MSC) d and bone marrow-derived macrophages (BMM) have propierties of tissular regeneration. Objectives: Determinated genetic expression of IL-4, Wnt6, VEGF and bFGF, in primary cellular cultures of MSC derivated of adipose tissue and BMM of C57BL/6 mice, through real time PCR (qRT-PCR). Methods: By an in silico analysis, we created primers to evaluate the proposed molecules in the primary cellular cultives, with qRT-PCR and electrophoresis. Results and projections: IL-4 and Wnt6 were not expressed in the MSC and BMM samples. VEGF and bFGF were expressed by different cells, giving differential expression. In the future, the same samples should be analyzed in an inflammatory environment, especially VEGF and bFGF. Limitations: The number of molecules are limited and the expression of them is only in a genetic level.

IL-4-C-590T locus polymorphism and susceptibility to asthma in children: a meta-analysis

Abstract Objectives The study aimed to evaluate the link between the IL-4-C590T polymorphism and asthma susceptibility in children by meta-analysis. Sources The study collected all the case-control studies found in PubMed, Embase, CNKI, Wanfang, VIP, and other databases until September 2019. Stata v. 15.0 was used to conduct meta-analysis, calculate the combined OR and its 95% CI, and then conduct subgroup analysis. Summary of the findings Seven studies were included in the study, containing 860 cases and 810 controls. Relative to the C allele, the T allele at the IL-4-C590T locus was associated with susceptibility to asthma in children (OR = 1.45, 95% CI: 1.05-2.01). The results of ethnicity subgroup analysis showed that there was statistical significance, with OR = 1.61 (95% CI: 1.01-2.57) in the Asian population. In the dominant and recessive genetic models, the overall test and the Asian population subgroup analysis were statistically significant. In the homozygous model, there was statistical significance, but no statistical significance in heterozygous model. Conclusions The IL-4-C590T polymorphism was associated with asthma susceptibility, and T allele and TT genotype may increase the risk of asthma susceptibility in children, especially in the Asian population.

Mechanism of Total Saponins from Dioscoreae Nipponicae Rhizoma to Treat Gouty Arthritis by Regulating M1/M2 Polarization of Macrophages / 中国实验方剂学杂志

Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups： normal group， model group， total saponin group （160 mg·kg^-1）， celecoxib group （43.3 mg·kg^-1）， with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin（HE） staining. Immunohistochemistry method was used to detect the protein expression change of CD68， interleukin-4（IL-4）， inducible nitric oxide synthase （iNOS） and transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁）. Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling， and the disease was in the acute stage. On day 5， the inflammation was alleviated， and on day 8， the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue， and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd，5th and 8th day of administration （<italic>P</italic><0.01）. Compared with the model group， the total saponins group could reduce the expression of CD68 and iNOS （<italic>P</italic><0.05，<italic>P</italic><0.01）on the 3rd day of administration， and significantly reduced them expression on the 5th and 8th days （<italic>P</italic><0.01）. Compared with the normal group， IL-4 and TGF-<italic>β</italic>₁ expression were increased in the model group when the drug was given for three days（<italic>P</italic><0.01）. Total saponin group could enhance IL-4 expression（<italic>P</italic><0.05）and decreased the TGF-<italic>β</italic>₁ expression（<italic>P</italic><0.01）. Compared with normal group， the expression of IL-4 in the model group decreased on the 5th and 8th day of administration （<italic>P</italic><0.01）， and the expression of TGF-<italic>β</italic>₁ in the model group decreased on the 5th day of administration（<italic>P</italic><0.01）. Compared with the model group， the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic>₁ at 5 d and 8 d after administration （<italic>P</italic><0.01）. Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.

Effect of Bletillae Rhizoma Polysaccharide on PI3K/Akt in Gastric Tissue of GU Model Rats / 中国实验方剂学杂志

Objective::To investigate the effect of Bletillae Rhizoma polysaccharide on the expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) gene protein and its mediated cytokines interleukin-2 receptor (IL-2R) and interleukin-4 (IL-4) in gastric tissue of rats with gastric ulcer (GU). Method::Sixty SPF Wistar rats were randomly divided into blank group and model group.The GU model was replicated by direct acetic acid cauterization in model group.The GU model rats were randomly divided into five groups: model group, positive control group, and large, medium and small-dose Bletillae Rhizoma polysaccharide groups, with 10 rats in each group.Rats in blank group and GU model group were given 10 mL·kg-1·d-1 distilled water by gavage, rats in large, medium and small-dose groups were given 0.5, 0.25, 0.125 g·kg-1·d-1 Bletillae Rhizoma polysaccharide by gavage, while rats in positive control group were given 0.3 g·kg-1·d-1 ranitidine by gavage for 15 days.Serum nitric oxide (NO) content, pepsinase activity and cytokines IL-2R and IL-4 levels in rats of each group were measured by enzyme-linked immunosorbent assay (ELISA), PI3K and Akt mRNA expressions were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), and PI3K and Akt protein expressions were detected by Western blot. Result::Compared with the blank group, the contents and gene expressions of cytokines IL-2R and IL-4 in gastric tissue were significantly increased, and the PI3K and Akt genes and protein expressions were significantly increased, with statistical significance (P<0.01). Compared with GU model group, the content and gene expressions of IL-2R and IL-4 in large, medium and small-dose Bletillae Rhizoma polysaccharide groups were decreased significantly, and the PI3K and Akt gene and protein expressions were decreased significantly in large-dose Bletillae Rhizoma polysaccharide group, while those in large and medium-dose Bletillae Rhizoma polysaccharide groups were decreased significantly (P<0.05, P<0.01). Conclusion::Bletillae Rhizoma polysaccharide can protect gastric mucosa by down-regulating PI3K and Akt gene and protein expressions and inhibiting abnormal secretion of cytokines IL-2R and IL-4.

Effect of electroacupuncture combined with pill on sex hormone and Th2 cytokines in patients of decreased ovarian reserve function with liver-kidney deficiency / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) combined with pill on clinical symptoms, levels of serum sex hormone and Th2 cytokines in patients of decreased ovarian reserve function (DOR) with liver-kidney deficiency, and to compare the efficacy between EA combined with pill and pill alone.@*METHODS@#Sixty patients with DOR were randomly divided into an observation group (30 cases, 2 cases dropped off) and a control group (30 cases, 1 case dropped off). The patients in the control group were treated with pill, 1 pill each time, 3 times a day. Based on the treatment of the control group, the patients in the observation group were additionally treated with acupuncture at Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Zigong (EX-CA 1), Zusanli (ST 36), Sanyinjiao (SP 6), Taixi (KI 3) and Taichong (LR 3); EA was applied at bilateral Zusanli (ST 36) and Sanyinjiao (SP 6), with continuous wave, in frequency of 20 Hz and current intensity of 1 to 4 mA, for 20 min. The treatment was given 3 times a week. All the patients terminated treatment during menstrual period, and the treatment was given for 3 continuous menstrual cycles. The menstrual condition score and systemic symptom score were compared between the two groups before and after treatment. The levels of serum sex hormones on 2nd to 3rd day of menstruation, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2), and the serums levels of interleukin (IL) -4 and IL-10 secreted by Th2 cytokines were compared between the two groups before and after treatment.@*RESULTS@#After the treatment, the menstruation condition scores and systemic symptom scores in the two groups were reduced (<0.05), and the scores in the observation group were lower than those in the control group (<0.05). After the treatment, the levels of serum FSH, LH and FSH/LH were reduced (<0.05), and the E2 levels were increased in the two groups (<0.05), and the levels of FSH, LH in the observation group were lower than those in the control group (<0.05), and the E2 level was higher than that in the control group (<0.05). After the treatment, the levels of serum IL-4 and IL-10 in the two groups were increased (<0.05), and the levels of IL-4 and IL-10 in the observation group were higher than those in the control group (<0.05).@*CONCLUSION@#EA combined with pill could significantly improve menstruation, systemic symptoms and serum sex hormone levels in patients of decreased ovarian reserve function with liver-kidney deficiency, which may restore ovarian function by up-regulating the expression of Th2 cytokines.

New insights into roles of basophils in initiating T helper type 2 immunity / 中草药·英文版

Allergic diseases, mainly mediated by T helper type 2 (Th2) immunity, have become a worldwide public health problem. Traditional Chinese medicine (TCM) has long been used in treating and preventing allergic symptoms. As the new target of anti-allergy TCM, basophils, after approximately 140 years since their discovery, are just now gaining respect as important contributors in the pathogenesis underlying allergic inflammation and disease. In addition to their role as effector cells, basophils can release early IL-4, migrate from circulatory system into draining lymph nodes, present antigen to naive CD4+ T cells, and promote the differentiation of Th2 cells. Herein, we briefly summarized the recent research advances of the essential contributions of basophils in the initiation of Th2 immune responses.

Association of Serum Vitamin D, IL-4 Levels and Vitamin D Receptor Gene Polymorphism in Coronary Artery Disease with and Without Type 2 Diabetes Mellitus

The aim of this study was to investigate the association of serum vitamin D, IL-4 levels and vitamin D receptor gene polymorphism in coronary artery disease with and without type 2 diabetes mellitus. Methods: The study was conducted in Department of Medicine and Department of Biochemistry, Maulana Azad Medical College and Lok Nayak Hospital, New Delhi. It involved two groups of patients suffering from CAD with type 2 diabetes mellitus (n =40) and CAD without type 2 diabetes mellitus (n =40). Blood sample was collected from all subjects using all aseptic precautions. The levels of serum 25-hydroxy Vitamin D were measured by Electrochemiluminescence Immunoassay. Expected normal serum values considered was 14-80ng/ml. Serum IL-4 had been measured by using commercially available ELISA kit provided by GEN- PROBE Diaclone, France. Expected normal serum value considered was < 98pg/ml. Results: The mean age of patients in different study groups were CAD with DM, 59.15± 9.31 years and CAD without DM, 58.1±9.51 years. Mean vitamin D levels were 18.6±8.3 ng/ml in CAD with DM and 23.4±9 ng/ml in CAD without DM. Mean IL-4 levels were 1.31±0.27pg/ml in CAD with DM group, 1.21±0.29pg/ml in CAD without DM group. The FF genotype of vitamin D receptor gene was present in 47.5 % of CAD with DM patients and 35 % of CAD without DM patients. The Ff genotype was present in 37.5 % of CAD with DM patients and 52.5 % of CAD without DM patients. The ff genotype was reported in 15 % of CAD with DM patients and 12.5 % of CAD without DM patients. Allele F of Vitamin D receptor gene constituted 66 % of total gene pool in CAD with DM patients and 61 % in CAD without DM patients. No significant association was observed with respect to the VDR FokI genotypes and cardiovascular outcomes. Conclusion: Serum Vitamin D levels were decreased in both groups of patients, more significantly decreased in the presence of DM in CAD patients. Serum IL-4 levels were significantly higher in CAD with DM group as compared to CAD without DM group. No associations could be found between Vitamin D receptor gene FokI polymorphism and risk of CAD in diabetic and non-diabetic individuals. No significant correlation was found between vitamin D and IL-4 levels in the patients of both groups. The association between VDR FokI polymorphism, vitamin D and inflammatory markers needs to be further explored in diabetic CAD patients.

Mechanism of Th2 related inflammatory factors in the pathogenesis of eczema / 药学实践杂志

Eczema is a common clinical inflammatory skin disease characterized by severe itching and recurrent attacks.Its etiology is complex, including a variety of internal and external factors.The main mechanism is related to the imbalance of Th2immune response, and related inflammatory factors play an important role.Epithelial-derived factor like thymic stromal lymphopoietin and interleukin-33could trigger Th2immune imbalance and promote the secretion of type inflammatory factors such as interleukin-4, interleukin-5, and interleukin-13.These inflammatory factors will further induce eosinophilic granulocytosis and IgE formation.In this paper, the research progress of mechanism of type 2related inflammatory factors in eczema were reviewed, which provided the great significance for treatment.

Clinical Efficacy of Modified Xiaofengsan and Its Effect on Cytokines in Persistent Asthma Patients with Wind-asthma Pattern / 中国实验方剂学杂志

Objective: To observe the clinical efficacy of modified Xiaofengsan and its effect on cytokines in persistent asthma patients with wind-asthma pattern. Method: The 120 eligible patients with chronic and persistent asthma were randomly divided into control group (60 cases) and treatment group (60 cases). Patients in control group received the budesonide/formoterol (160 μg/4.5 μg, 2 inhales/time, bid). Patients in treatment group additionally received the modified Xiaofengsan combined with budesonide/formoterol (160 μg/4.5 μg). The treatment course was 1 month in both groups. The clinical efficacy of the two groups was observed. Before and 3 months after treatment, asthma control test (ACT) and the average times of acute exacerbation in 6 months before and after treatment between two groups were observed; percentage of forced expiratory volume in first second and its predicted value (FEV1%), variety ratio of Peak expiratory flow (PEF), eosinophilic granulocyte (EOS) count in peripheral blood and the levels of interleukin-4(IL-4), interleukin-6(IL-6) and interleukin-13(IL-13) in serum were measured before and after treatment respectively. In addition, the safety of the two groups was evaluated. Result: The total effective rate of the treatment group was higher than that of the control group (P1% in treatment group were higher(PPConclusion: Modified Xiaofengsan combined with budesonide/formoterol could relieve symptoms of asthma, improve pulmonary function and lower the levels of IL-4, IL-6 and IL-13 in serum.

Expression of serum Th1/Th2 cytokines in different stages of hepatitis B and its clinical significance / 国际检验医学杂志

Objective To investigate the expression levels of IFN-γ, IL-4, IL-6 in the plasma of HBV chronic patients.To acknowledge the correlation between the immune function of T-cell with chronic hepatitis B virus.Methods The expression levels of IFN-γ, IL-4, IL-6 in the plasma were determined by ELISA in 42 cases of chronic hepatitis B (CHB group), 30 cases of cirrhosis (LC group), 30 cases of primary liver cancer (HCC group) and 30 cases of health control (NC group).Combine with the load of HBV DNA, HBeAg and ALT, the expression levels of IFN-γ, IL-4, IL-6 were re-analysed.Results The expression levels of IFN-γ, IL-4 and IL-6 in the researched groups were statistically significantly different.The expression levels of IFN-γ, IL-4, IL-6 in CHB, LC and HCC groups were higher than NC group with significance.The expression level of IL-4 in the CHB group was higher than LC and HCC groups with significance.The expression level of IL-6 in the HCC group was higher than CHB and LC groups with significance.Compared with the control group, the ratio of IFN-γ/IL-4 in CHB, LC and HCC groups were decreased with significance.There was significantly positive correlation between the expression level of IL-4 with the load of HBV DNA (r=0.757, P＜0.001).The expression level of IFN-γhad negative correlation between the load of HBV DNA (r=-0.786, P＜0.001) with the level of ALT (r=-0.713, P＜0.001) with significance.There was no significant correlation between HBeAg with the cytokines in the serum of patients with chronic HBV.Conclusion The results showed the immune function of T-cell play important role in chronic HBV patients.That would provide a good basis on CHB treatment.The expression levels of IL-6 were higher in HCC group with significance, which provided the direction for the deep study of liver cancer.