Involvement of PM2.5-bound protein and metals in PM2.5-induced allergic airway inflammation in mice

Abstract

Background: The aim of this study was to investigate the protein and trace element components of PM2.5 and their contribution to the allergic airway inflammation in BALB/c mice.

Methods: PM2.5, treated at high temperature and with a strong acid to hydrolyze any protein content and remove trace elements, was administered to BALB/c mice. Allergic airway inflammation was compared between the three groups (saline, pure PM2.5 and treated PM2.5) by evaluating airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid (BALF) cells, serum IgE, the mRNA of various cytokine (IL-4, IL-5, IL-13, eotaxin-1 and CXCL3), mucus protein mRNA (MUC5ac and MUC5b) and the filtration of inflammatory cells in the lung.

Results: The treatment of PM2.5 with a strong acid at a high temperature attenuated AHR, eosinophil percentage in BALF, mRNA levels of IL-13 and CXCL3 and peribronchial inflammation. On the contrary, the percentage of neutrophils in BALF, mRNA expression of MIP2α, EGFR, Nrf2, and TLR4 and 4-OH-2-nonenal levels in the lung was increased. Moreover, the treatment of the PM2.5 reduced PM2.5-bound proteins as well as the percentages of the trace elements in PM2.5 in the order Zn?>?Cu?> Pb?>?P?>?S?>?Mn?>?Fe?>?Ca?>?Ni, whereas the percentage of C, Si and Cl increased.

Conclusions: PM2.5 collected by of the cyclone system induced allergic airway inflammation in mice. PM2.5-bound proteins and acid-soluble metals may be involved in the pathogenesis of PM2.5-induced allergic airway inflammation.     

Urban particulate matter induces pro-remodeling factors by airway epithelial cells from healthy and asthmatic children

Abstract

Context: Chronic exposure to ambient particulate matter pollution during childhood is associated with decreased lung function growth and increased prevalence of reported respiratory symptoms. The role of airway epithelium-derived factors has not been well determined.

Objective: To determine if urban particulate matter (UPM) stimulates production of vascular endothelial growth factor (VEGF) and transforming growth factor-β₂ (TGF-β₂), and gene expression of mucin 5AC (MUC5AC) and interleukin-(IL)-8 by primary airway epithelial cells (AECs) obtained from carefully phenotyped healthy and atopic asthmatic school-aged children.

Methods: Primary AECs from 9 healthy and 14 asthmatic children were differentiated in air--liquid interface (ALI) culture. The apical surface was exposed to UPM suspension or phosphate buffered saline (PBS) vehicle control for 96?h. VEGF and TGF-β₂ concentrations in cell media at baseline, 48 and 96?h were measured via ELISA. MUC5AC and IL-8 expression by AECs at 96?h was measured via quantitative polymerase chain reaction.

Results: Baseline concentrations of VEGF, but not TGF-β₂, were significantly higher in asthmatic versus healthy cultures. UPM stimulated production of VEGF, but not TGF-β₂, at 48 and 96?h; the magnitude of change was comparable across groups. At 96?h there was greater MUC5AC and IL-8 expression by UPM exposed compared to PBS exposed AECs.

Conclusions: Induction of the pro-remodeling cytokine VEGF may be a potential mechanism by which UPM influences lung function growth in children irrespective of asthma status. Respiratory morbidity associated with UPM exposure in children may be related to increased expression of MUC5AC and IL-8.     

基于IL-13/STAT6信号通路研究祛风宣痹方对哮喘气道黏液高分泌的影响

目的：探讨祛风宣痹方对哮喘小鼠气道黏液分泌及IL-13/STAT6信号通路的影响。方法：采用卵清蛋白（OVA）致敏建立小鼠哮喘模型。将24只Balb/c雌性小鼠随机分为对照组、哮喘组、祛风宣痹方组，祛风宣痹方组给予祛风宣痹方治疗，对照组和哮喘组给予等量生理盐水灌胃。采用肺组织HE染色、PAS染色观察肺组织病理形态变化以及气道黏液分泌，免疫组化实验观察黏蛋白MUC5AC、p-STAT6蛋白表达情况，ELISA实验检测小鼠血清中IL-13的含量。IL-13处理A549细胞模拟黏蛋白分泌病理模型（IL-13组），再加入祛风宣痹方进行干预（IL-13+祛风宣痹方组）；采用免疫荧光技术检测A549细胞中MUC5AC和p-STAT6的表达情况。结果：体内实验发现，与哮喘组相比，祛风宣痹方组小鼠肺组织中炎症细胞浸润及黏液高分泌情况均有明显改善，气道表面细胞MUC5AC、p-STAT6蛋白表达有所降低（P < 0.05）；小鼠血清中IL-13含量也明显减少（P < 0.01）。体外实验发现，与对照组相比，IL-13组A549细胞中MUC5AC及p-STAT6蛋白表达明显升高（P < 0.01），而IL-13+祛风宣痹方组细胞中MUC5AC及p-STAT6蛋白表达相对于IL-13组呈现明显下降趋势（P < 0.05）。结论：祛风宣痹方可减轻哮喘气道黏液高分泌，其作用机制可能与抑制IL-13/STAT6信号通路相关。     

Effects of naphthoquinone on airway responsiveness in the presence or absence of antigen in mice

We have recently demonstrated that naphthoquinone (NQ), one of extractable chemical compounds of diesel exhaust particles (DEP), enhances antigen-related airway inflammation with goblet cell hyperplasia in mice (Inoue et al. in Eur Respir J 209(2):259–267, 2007). Further, NQ has enhanced lung expressions of interleukin (IL)-4 and IL-5. However, the effects of NQ on other cardinal features of asthma have not been completely investigated. The aim of the present study was to evaluate the effects of NQ on airway responsiveness on the model. Vehicle, NQ, ovalbumin (OVA), or NQ + OVA was administered intratarcheally to ICR mice for 6 weeks. Twenty-four hours after the last instillation, lung histology, lung functions such as total respiratory system resistance (R) and Newtonian resistance (R _n), and protein level of IL-13 and mRNA level for MUC5AC in the lung were examined. Repetitive exposure to NQ aggravated antigen-related lung inflammation. NQ alone enhanced R and R _n as compared to vehicle without statistical significance. OVA alone or NQ plus OVA showed increases in R and R _n, which was prominent in NQ plus OVA (P < 0.05 vs. vehicle). Combined exposure to NQ and OVA elevated the levels of IL-13 and MUC5AC in the lung as compared with exposure to NQ or OVA alone. These results indicate that NQ can enhance airway hyperresponsiveness in the presence or absence of an antigen. Also, amplified lung expressions of IL-13 and MUC5AC might partly contribute to the deterioration of asthma features by NQ.     

Zingiber officinale ameliorates allergic asthma via suppression of Th2-mediated immune response

Abstract

Context: Ginger has been used commonly in the traditional system of medicine for the treatment of respiratory disorders.

Objective: The present study investigates the immunosuppressive activity of ginger by using the mouse model of ovalbumin-induced allergic asthma.

Materials and methods: Treatment with ethanol extract (500?mg/kg) and aqueous extract (720?mg/kg) of rhizomes, and methylprednisolone (5?mg/kg) was initiated 1 week after second sensitization of mice with ovalbumin and continued for 7?d. RT-PCR followed by gel electrophoresis and ELISA were used for the evaluation of mRNA expression levels and protein levels of Th2 type markers, respectively. Lung tissue histopathology was conducted by using H&E and PAS staining.

Results: We observed significant reduction in goblet cell hyperplasia (0.83?±?0.17 and 1.0?±?0.26), infiltration of inflammatory cells in airways (0.67?±?0.33 and 1.0?±?0.37), and edema with vascular congestion (1.0?±?0.26 and 1.2?±?0.17) by both ethanol and aqueous extracts, respectively. A highly significant reduction of total and differential count of eosinophils and neutrophils in BALF, and eosinophil count in blood were also observed. Both extracts significantly inhibited Th2-mediated immune response, which is evident by a decrease in mRNA expression levels of IL-4 and IL-5. Protein levels of IL-4 and IL-5 in BALF, along with total serum IgE levels, were also significantly suppressed by both extracts.

Discussion and conclusion: Our study validated the traditional use of ginger in respiratory disorders and suggests that ginger reduces allergic airway inflammation, possibly by the suppression of Th2-mediated immune response.     

血红素氧合酶-1对哮喘小鼠模型IL-25和MUC5AC表达的影响

摘要：目的　探讨血红素氧合酶-1（HO-1）对哮喘小鼠气道炎症的调节作用及其机制。方法　将28只健康雌性Balb/c小鼠根据随机数字表法分为4组：对照组、哮喘组、血晶素组、福多司坦组,每组7只。哮喘组、血晶素组和福多司坦组小鼠于第1天和第14天腹腔注射20 μg卵清蛋白（OVA）+500 μg Al（OH）3+0.2 mL PBS致敏,于第25~28天每天雾化吸入1次OVA激发气道高反应制备小鼠哮喘模型,对照组以等量生理盐水替代,血晶素组致敏前1、2 d,致敏后12、13、23、24、27 d腹腔注射血晶素75 μmol/kg,福多司坦组分别于致敏前25、26、27 d灌胃给药福多司坦（200 mg/kg）,末次激发后24 h内处死小鼠并收集血液、肺组织。HE染色观察肺组织形态学改变,免疫组化检测肺组织黏蛋白5AC（MUC5AC）的表达,酶联免疫吸附测定（ELISA）法检测肺组织中MUC5AC和血清中白细胞介素（IL）-25的水平,并对MUC5AC和IL-25进行相关性分析。结果　哮喘组肺组织可见大量炎症细胞浸润、气管黏膜增厚、黏膜下水肿,血晶素组和福多司坦组可见少量炎症细胞浸润及渗出,病理改变较哮喘组轻。与对照组相比,哮喘组、血晶素组、福多司坦组MUC5AC和IL-25的表达水平明显升高（P＜0.05）;与哮喘组相比,血晶素组和福多司坦组MUC5AC和IL-25降低（P＜0.05）,且血晶素组较福多司坦组降低更明显（P＜0.05）。MUC5AC与IL-25的表达水平呈正相关（r=0.932,P＜0.01）。结论　HO-1可下调哮喘小鼠MUC5AC和IL-25的表达,减轻气道炎症和黏液高分泌。     

罗格列酮对内毒素诱导气道MUC5AC表达的调控

目的探讨过氧化物酶体增殖物激活受体γ（PPARγ）激动剂罗格列酮对内毒素所致大鼠气道MUCSAC表达的影响及其机制。方法将36只雄性SD大鼠随机分为生理盐水对照组（A组）、罗格列酮对照组（B组）、脂多糖组（LPS组）（C组）、以及低、中、高剂量罗格列酮组（D组、E组和F组）。采用酶联免疫吸附测定（ELISA）法检测支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF—α）、白细胞介素-1β（IL-1β）和IL-10浓度。免疫组化染色及实时荧光定量逆转录-PCR检测气道肺组织MUC5AC蛋白及mRNA表达。结果与生理盐水对照组比较,LPS组BALF中TNF-α、IL-1β和IL-10浓度,以及气道肺组织MUC5AC蛋白和mRNA表达增加（P〈0．01）;TNF-α、IL-1β与MUC5AC mRNA表达呈正相关（r分别为0．851,0．803。P〈0．05）;IL-10浓度与MUC5AC mRNA表达呈负相关（r为-0．812,P〈0．05）。给予低、中、高剂量罗格列酮干预后,D组、E组和F组BALF中TNF—α浓度及MUC5AC蛋白、mRNA表达均逐渐降低,IL-10水平逐渐升高,各组间差异有统计学意义（P均〈0．05）。结论罗格列酮对内毒素诱导的气道黏液高分泌有拮抗作用,其具体机制可能与调节炎症反应,下调气道MUC5AC转录有关。     

The role of p38 MAPK in acute paraquat-induced lung injury in rats

Abstract

Context: Paraquat (PQ; 1,1'-dimethyl-4,4'-bipyridinium dichloride) is highly toxic and accounts for a large proportion of the herbicide poisonings seen in clinic. The major cause of mortality is respiratory failure. The p38 mitogen-activated protein kinase (MAPK) signal transduction pathway coordinates various cellular stress responses that have been shown to participate in the pathogenesis of PQ-induced lung injury.

Objective: To evaluate the effect of the specific p38 MAPK inhibitor SB203580 on PQ-induced lung injury and cytokine secretion.

Methods: In groups of 24, rats were treated with PQ, PQ and SB203580 (SB?+?PQ), SB203580 alone (SB) or normal saline (control group). Six rats from each group were euthanized at 1, 3, 5 or 7?d. Pathology of lung specimens was scored through hematoxylin and eosin staining. Edema in the lung was quantified from wet-to-dry weight ratios. p38 and p-p38MAPK proteins were measured via electrochemiluminescent Western blots. tumor necrosis factor (TNF)-alpha and interleukin-1 beta (IL-1β) concentrations in lung specimens and bronchoalveolar lavage fluid (BALF) were quantified via enzyme-linked immunosorbent assay.

Results: The mortality rate of the SB?+?PQ group (16.7%) was significantly lower than that of the PQ group (33.3%; p?<?0.05). The PQ group had significantly higher pulmonary histology scores, wet-to-dry weight ratios and phosphorylated p-p38 MAPK levels, as well as higher IL-1β and TNF-alpha levels in BALF and lung tissues, that did the SB?+?PQ and control groups (p?<?0.05, all).

Conclusion: The data suggest that the p38 MAPK signaling pathway has an important role in regulating the production of IL-1β and TNF-alpha in PQ-induced lung injury in rats.     

CCR3 monoclonal antibody inhibits airway eosinophilic inflammation and mucus overproduction in a mouse model of asthma

AIM: To explore the effect of a rat anti-mouse CC-chemokine receptor-3 (CCR3) monoclonal antibody (CCR3 mAb) on airway eosinophilia and mucus overproduction in asthmatic mice. METHODS: An asthma model was sensitized and challenged by ovalbumin (OVA) in male C57BL/6 mice. Asthmatic mice were given dual administration (intraperitoneal injection and aerosol inhalation) of CCR3 mAb or nonspecific rat IgG (ns-IgG). The number of total and differential inflammatory cells in the bronchial alveolar lavage fluid (BALF) was counted. Eosinophils number, the goblet cell percentage (GCP) and airway mucus index (AMI) were measured in the lung tissues. Interleukin (IL)-5 levels in the BALF were examined. The expression of MUC5AC and the epidermal growth factor receptor (EGFR) mRNA in the lung tissues was detected by semi-quantitative RT-PCR. The results were compared among the groups. RESULTS: CCR3 mAb significantly suppressed the increased eosinophils in the BALF and lung tissues in OVA-challenged mice compared with ns-IgG-treated mice. IL-5 levels in the BALF in CCR3 mAb and ns-IgG administration mice exhibited no obvious changes relative to OVA-challenged asthmatic mice. CCR3 mAb reduced the increased GCP and AMI after OVA challenge and decreased the enhanced expression of MUC5AC and EGFR mRNA in lung tissues in asthmatic animals. CONCLUSION: CCR3 mAb can significantly inhibit airway eosinophilia and mucus overproduction in asthmatic mice. Blockage of CCR3 may represent a new strategy to asthma therapy.     

Reversing the polarization of tumor-associated macrophages inhibits tumor metastasis

ObjectiveThe M2 phenotype is dominant in tumor associated macrophages (TAM), and plays a key role in promoting tumor growth, invasion and metastasis. Converting TAM polarization from M2 to M1 may contribute to eliciting anti-tumor-specific immune responses and inhibiting tumor metastasis. In this study, the effect of reversing the polarization of TAM on tumor metastasis was investigated.MethodsPeritoneal macrophages were obtained from BABL/c mice, and M2 polarization was induced by IL-4. In an in vivo experiment, BABL/c mice were transplanted with 4 T1 tumor cells. In vitro and in vivo experimental studies, M2 macrophage polarization was reversed with CpG-DNA or CpG-DNA combined with anti-IL-10R Ab. CD68, MHCII and FRβ molecular expression in macrophages were examined with immunofluorescence staining. The mRNA expression of IL-2, IL-6, IL-13, VEGF and MMP-9 were detected with RT-PCR. VEGF and MMP-9 protein expression of tumors in situ was measured by western blot assay. Lung-metastasis of the tumor was observed and assessed by micro-CT.ResultsCpG-DNA and CpG-DNA combined with anti-IL-10R Ab could promote MHCII, IL-2, IL-6 and IL-13 molecular expression, and suppress the expression of FRβ, MMP-9 and VEGF, in both freshly isolated peritoneal macrophages and M2 macrophages. In the CpG-DNA combined with anti-IL-10R Ab injecting group, the percentage of CD68⁺ MHCII⁺ cells were significantly higher than that of CD68⁺ FRβ⁺ cells (P < 0.05). This was distinct from the result of the control group, which CD68⁺ FRβ⁺ was higher than CD68⁺ MHCII⁺ cells (P < 0.01). Furthermore, VEGF-A and MMP-9 level in primary tumor tissues in the experimental group was significantly lower (P < 0.01), compared to the control group. Moreover, the number of detectable lung-metastasis foci was significantly lower in the experimental group than in the control group (P < 0.05).ConclusionReversing the polarization of TAM from M2 to M1 phenotype can inhibit tumor metastasis.     

Human albumin augmented airway inflammation induced by PM2.5 in NC/Nga mice

The synergic allergic inflammatory effects of particulate matter (PM) 2.5 and human albumin were investigated in NC/Nga mice, which are hypersensitive to mite allergens. PM2.5 or PM2.5 plus human albumin with aluminum oxide was injected twice intraperitoneally for sensitization. After 7 days, PM2.5 or PM2.5 plus human albumin was administered five times intranasally to mice for further sensitization. Subsequently, PM2.5 was administered as a challenge on the 11th day. On the 12th day, mice were examined for airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid (BALF) cell count, mRNA expression of Th₁, Th₂ cytokines, chemokines, and mucus proteins (MUC5AC and MUC5B) in the lung tissue and histopathology. Although PM2.5 or human albumin alone did not induce allergic airway inflammation, simultaneous inoculation of PM2.5 and human albumin‐induced airway inflammation showing increase in AHR, total BALF cell numbers, mRNA levels of IL‐13, eotaxin 1, eotaxin 2, and MUC5AC, and anti‐IG against human serum albumin. Inflammation was observed around the bronchus in PM2.5 plus human albumin‐induced lungs. These results demonstrate that PM2.5 can induce allergic airway inflammation through the synergistic action with human albumin in NC/Nga mice.     

Differential expression of two genes Oct‐4 and MUC5AC associates with poor outcome in patients with gastric cancer

Gastric cancer (GC) is the most frequent leading cause of cancer‐associated mortality worldwide that is linked to poor prognosis due to the lack of appropriate biomarkers. Our aim was to evaluate the MUC5AC and Oct‐4 expression levels in GC and to assess their association with clinical factors. Immunohistochemical analysis (IHC) and qRT‐PCR were performed in GC patients to examine the MUC5AC and Oct‐4 expression levels. The mRNA level of MUC5AC was significantly decreased in tumour tissues compared with non‐cancerous tissues (1.11 ± 0.69 vs 3.7 ± 0.71; P = .024). On the other hand, Oct‐4 mRNA level was upregulated in tumour tissues as compared to normal tissues (2. 86 ± 0.78 vs 0.87 ± 0.54; P = .0015). Decreased expression of MUC5AC was detected in 27 patients (67.5%), while high to moderate expression levels were observed in 13 cases (32.5%), but in normal tissues the expression levels of MUC5AC were increased (P = .001). The decreased expression of MUC5AC was associated with aggressive tumour characteristics, such as TNM stage (P = .023), histologic type (P = .012) and lymph node metastasis (P = .001). High expression of Oct‐4 was detected in 24 tumour tissues (60%), while 16 cases (40%) showed low expression level. Increased Oct‐4 expression was correlated with clinicopathological characteristics such TNM stage (P = .002), histologic type (P = .008) and lymph node metastasis (P = .001). Our results showed that high Oct‐4 expression and the reduction of MUC5AC expression may be involved in the progression and an unfavorable prognosis of GC.     

The effects of chrysophanol on ovalbumin (OVA)-induced chronic lung toxicology by inhibiting Th17 response

Chrysophanol (CH), extracted from plants of Rheum genus, possesses various pharmacological effects including anti-inflammatory activity. The purpose of the present study was to evaluate the protective effects and the underlying mechanisms of CH on ovalbumin (OVA)-induced asthma in mice. Fifty mice were randomly assigned to five experimental groups: control group, model group, dexamethasone (2?mg/kg) group and CH (5 and 10?mg/kg) groups. The number of eosinophil cells and the production of interleukin-6 (IL-6), IL-1β, IL-17?A and tumor necrosis factor-α in bronchoalveolar lavage fluid (BALF) were measured. In addition, pulmonary histopathology, airway resistance (Raw), T-helper17 (Th17) cells frequency and RORγt expression were evaluated. Our study demonstrated that CH effectively decreased eosinophil count and inflammatory cytokines production in BALF. In addition, treatment with CH significantly inhibited the Raw, Th17 percentage and RORγt expression in OVA-induced animals compared with those in model group. Histological studies also demonstrated that CH significantly suppressed OVA-induced eosinophilia in lung tissue compared with model group. Our findings supported that CH can prevent allergic asthma in the mouse model.     

The role of Act1, a NF-κB-activating protein,in IL-6 and IL-8 levels induced by IL-17 stimulation in SW982 cells

Abstract

Context: Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic inflammation in the synovial membrane of affected joints. It has been shown that several kinds of cytokine were increased in synovial fluid, while the underlying mechanism remains poorly understood.

Objectives: NF-κB activator 1 (Act1) is a recently identified protein binding to the IκB kinase complex. Our study aimed to investigate the expression of Act1 induced by cytokine IL-17 stimulation in SW982 cells.

Materials and methods: The human synovial sarcoma cell line SW982 and primary cultured RA fibroblast-like synovial cells were used. RT-PCR and Western blot assays were selected to investigate the genetic and protein expression of Act1. Additionally, four independent Act1 small interfering RNA (siRNA) oligonucleotides were designed and obtained according to the GenBank cDNA, the sequence of Act1 (Traf3ip2). Finally, enzyme-linked immunosorbent assay (ELISA) double antibody sandwich was used to assay supernatant IL-6 and IL-8 concentrations.

Results: The Act1 mRNA expression level increased significantly after stimulation with IL-17 (5–100?ng/ml) in SW982 cells. Additionally, the level of Act1 mRNA expression correlated positively with the concentration of IL-17 (p?<?0.01). IL-17 induced IL-6 and IL-8 in SW982 cells was in a concentration- and time-dependent way. Furthermore, ELISA assay revealed that IL-17 (20?ng/ml) significantly increased IL-6 (1927.4?±?288.77 versus 786.5?±?172.42?ng/ml, p?<?0.01) and IL-8 levels (984.8?±?95.09?ng/ml versus 307.1?±?90.83?ng/ml, p?<?0.01) compared with control group after stimulation for 24?h. However, transfection of Traf3ip2 siRNA markedly decreased IL-6 (995.9?±?115.30?ng/ml versus 1816.1?±?273.27?ng/ml, p?<?0.01) and IL-8 levels (575.6?±?65.96?ng/ml versus 929.4?±?124.39?ng/ml, p?<?0.01) compared to transfection negative control. These findings suggested that IL-6 and IL-8 level induced by IL-17 in SW982 cells could be reversed by down-regulation of Act1 expression level with Traf3ip2 siRNA.

Conclusion: Our results suggested that Act1 might play a key role in the pathophysiology and the treatment of RA.     

Protective effects of caffeoylxanthiazonoside isolated from fruits of Xanthium strumarium on sepsis mice

Abstract

Context: The fruit of Xanthium strumarium L. (Asteraceae) has been used for the treatment of various inflammatory diseases.

Objective: This study investigates the protective effect of caffeoylxanthiazonoside (CYXD) isolated from fruits of X. strumarium on sepsis mice in vitro and in vivo.

Materials and methods: Cecal ligation and puncture (CLP) operation was used to establish the sepsis mice model, and sham mice were also performed. CYXD was administered by intraperitoneal injection (10, 20, and 40?mg/kg/d), then the survival rate was measured in 96?h. Additionally, sepsis mice were induced by injection LPS (2?mg/kg); CYXD was administered by intraperitoneal injection (10, 20, and 40?mg/kg/d), then mice were sacrificed, and serum levels of TNF-α and IL-6 were determined by ELISA assay. Furthermore, the ability of CYXD to neutralize LPS was measured by using the LAL test, and expressions of TNF-α, IL-6 were determined by using real-time fluorogenic PCR.

Results: Results indicated that CYXD significantly elevated survival rates of sepsis mice induced by CLP (p?<?0.05) with survival rates of 35%, 45%, and 65%. Furthermore, the LPS level was decreased obviously by CYXD (1, 2, and 4?mg/L) (p?<?0.05). Additionally, CYXD (10, 20, and 40?mg/kg) can not only significantly decrease TNF-α and IL-6 levels induced by LPS in mice's serum (p?<?0.05), but also inhibit mRNA expressions of TNF-α and IL-6 induced by LPS in RAW 264.7 cells at doses of 20, 40, and 80?μg/mL (p?<?0.05).

Conclusion: Our study demonstrated that CYXD has significant protective effects on sepsis mice.     

降钙素原黏蛋白MUC5AC联合简化临床肺部感染评分对呼吸机相关性肺炎的早期诊断价值

目的 探讨降钙素原（PCT）、黏蛋白MUC5AC联合简化肺部感染评分（CPIS）对呼吸机相关性肺炎（VAP）的早期诊断价值。方法 选取2018年10月至2019年9月在安徽医科大学附属安庆医院重症医学科接受机械通气治疗的78例气管插管患者作为研究对象,依据VAP诊断标准,将其分为VAP组（42例）与非VAP组（36例）。比较两组患者机械通气48 h后血清PCT、黏蛋白MUC5AC及简化CPIS评分的差异,引用受试者工作特征（ROC）曲线评价上述指标对VAP的早期诊断价值。结果 VAP组患者黏蛋白MUC5AC水平为（107.21±16.36）ng/mL、PCT为（2.46±1.09）ng/mL、简化CPIS评分为（6.37±2.24）分,均高于非VAP组,差异均有统计学意义（P<0.05）。PCT、黏蛋白MUC5AC联合简化CPIS评分对早期诊断VAP的ROC曲线下面积、敏感度和特异度分别为0.962、93.32%和86.78%。结论 血清PCT、黏蛋白MUC5AC及简化CPIS评分在VAP患者中均明显增高,黏蛋白MUC5AC与简化CPIS评分呈正相关,3项指标联合分析在VAP早期诊断中具有较高的价值。     

The effect of vitamin E on tracheal responsiveness and lung inflammation in sulfur mustard exposed guinea pigs

Pulmonary complications of sulfur mustard (SM) range from mild respiratory symptoms to even severe bronchial stenosis. In the present study, the protective effect of vitamin E on tracheal responsiveness (TR) and lung inflammation of SM-exposed guinea pigs were examined. Guinea pigs were exposed to ethanol (control group), 40?mg/m³ inhaled SM and ethanol vehicle (sulfur mustard exposed (SME) group), SME treated with vitamin E (SME + E), SME with dexamethasone (SME + D) and both drugs (SME + E + D), (n?=?8 for each group). TR to methacholine, total and differential white blood cell (WBC) count of lung lavage and serum cytokines were evaluated 14 days post-exposure. TR, WBC, interleukin 4 (IL-4), interferon gamma (INF-γ), eosinophil, and monocyte levels in SME guinea pigs were significantly higher, but lymphocyte was lower than those of controls (P?<?0.05 to P?<?0.001). TR, IL-4, and eosinophil levels in SME + E, SME + D and SME + E + D, INF-γ in SME + E and SME + E + D and WBC in SME + E were significantly decreased compared to that of the SME group (P?<?0.01 to P?<?0.001). In addition, the TR of SME + D + E was significantly higher than that of SME + E (P?<?0.01) and SME + D (P?<?0.05) groups. The results showed a preventive effect of vitamin E, dexamethasone and their combination on TR and lung inflammation in SME guinea pigs.     

大环内酯类抗生素对慢性气道炎症黏液高分泌的干预机制

目的:探讨大环内酯类抗生素对慢性气道炎症黏液高分泌的干预机制.方法:利用大鼠慢性支气管炎模型,对其进行3种大环内酯类抗生素的持续给药干预,采用底物检测法、 ELISA法和原位杂交技术分别检测支气管肺泡灌洗液(BALF)中性粒细胞弹性蛋白酶(NE)活性、气道黏蛋白5AC(MUC5AC)含量和 MUC5AC mRNA的表达.结果:红霉素、克拉霉素、阿奇霉素干预后各指标与对照组比较,均具有显著性差异(P<0.01),但阿奇霉素的作用又弱于红霉素和克拉霉素(P<0.05).结论:大环内酯类抗生素的黏液抑制性调理作用可能是由于其内在化于中性粒细胞抑制 NE释放而间接实现的.     

Effect of Aegle marmelos leaf extract on N-methyl N-nitrosourea-induced hepatocarcinogensis in Balb/c mice

Abstract

Context and objective: Tobacco smoke and nitrostable foods containing N-methyl N-nitrosourea (MNU) are among the primary causes of liver cancer. To substantiate the beneficial claims ascribed to Aegle marmelos (L.) Corrêa (Rutaceae), the hepatoprotective potential of its leaf extract was studied using an MNU-induced hepatocarcinogenesis model in Balb/c mice.

Materials and methods: After dose selection, 40 mice were randomly assigned to 4 groups: I (control), II (intraperitoneally (i.p.) primed with 50?mg/kg MNU), III (100?mg/kg A. marmelos hydroalcoholic extract (HEAM) i.p.) and IV (MNU?+?HEAM, i.p.). Inflammatory (IL-1β, IL-6), anti-inflammatory (IL-4) cytokine expression, apoptosis (Bcl-2) and tumor-related (p53, c-jun) genes were assessed at mRNA level. HEAM effects on hematological parameters were examined.

Results and discussion: HEAM treatment decreased IL-1β, IL-6, Bcl-2 and c-jun respectively expressions by 90, 25, 53 and 30%, respectively. p53 and IL-4 expression was up-regulated by 1.5- and 2-fold. MNU decreased hemoglobin concentration (25%), lymphocyte count (42%) and increased leukocyte (100%), platelet (4-fold), neutrophil (43%), monocyte (10-fold) and eosinophil (10-fold) counts in Group II mice while HEAM modulated the same parameters by ?7%, ?21%, +24%, +3-fold, +12%, +3-fold and +4-fold, respectively, in MNU-induced mice compared to control. HEAM protective effect was confirmed by Raman spectroscopy where the MNU-induced peak at 1252?cm^?1 was normalized. DNA fragmentation data suggest apoptosis as one of the protective mechanisms of HEAM.

Conclusion: The hepatoprotective, anti-carcinogenic and immunomodulatory effects of A. marmelos extract indicate potential beneficial effects in cancer therapy.