Extent Of Inflammatory Cell Infiltration Research Articles

Amelioration of ovalbumin-induced lung inflammation in a mouse model by Trichinella spiralis novel cystatin.

Asthma, a chronic disease affecting humans and animals, has recently become increasingly prevalent and steadily widespread. The alternative treatment of asthma using helminth infections or helminth-derived immunomodulatory molecules (IMs) has been evaluated and demonstrated significant amelioration of disease severity index in vitro and in vivo. Trichinella spiralis, a parasitic nematode and its IMs, elicits a potential to relieve asthma and other immune-related disorders. In this study, we investigated the immunomodulatory function of recombinant T. spiralis novel cystatin (rTsCstN) in ameliorating acute inflammatory asthma disorders in a murine model. Female BALB/c mice were sensitized using intraperitoneal injection of ovalbumin (OVA)/alum and subsequently challenged with intranasal administration of OVA alone or OVA + rTsCstN for 3 consecutive days, producing OVA-induced allergic asthma models. To evaluate the therapeutic efficacy of rTsCstN, the inflammatory cells and cytokines in bronchoalveolar lavage fluid (BALF) and OVA-specific immunoglobulin E levels in serum were assessed. Histological alterations in the lung tissues were determined by hematoxylin and eosin (H&E) staining and eventually scored for the extent of inflammatory cell infiltration. The asthmatic mouse models challenged with OVA + rTsCstN demonstrated a significant reduction of eosinophils (p < 0.01), macrophages (p < 0.05), and cytokines tumor necrosis factor-α (p < 0.05) and interferon (IFN)-γ (p < 0.05) in BALF when compared with the mice challenged with OVA alone. However, the levels of interleukin (IL)-4 and IL-10 remained unchanged. Histological examination revealed that mice administered OVA + rTsCstN were less likely to have inflammatory cell infiltration in their perivascular and peribronchial lung tissues than those administered OVA alone. Recombinant T. spiralis novel cystatin demonstrated immunomodulatory effects to reduce severe pathogenic alterations in asthma mouse models, encouraging a viable alternative treatment for asthma and other immunoregulatory disorders in humans and animals in the future.

Integrated DNA Methylation and Transcriptomics Analyses of Lacrimal Glands Identify the Potential Genes Implicated in the Development of Sjögren's Syndrome-Related Dry Eye.

Sjögren's syndrome-related dry eye (SS-related dry eye) is an intractable autoimmune disease characterized by chronic inflammation of lacrimal glands (LGs), where epigenetic factors are proven to play a crucial role in the pathogenesis of this disease. However, the alteration of DNA methylation in LGs and its role in the pathogenesis of SS-related dry eye is still unknown. Here, we performed an integrated analysis of DNA methylation and RNA-Seq data in LGs to identify novel DNA methylation-regulated differentially expressed genes (MeDEGs) in the pathogenesis of SS-related dry eye. The DNA methylation and transcription profiles of LGs in NOD mice at different stages of SS-related dry eye (4-, 8-, 12- and 16 weeks old) were generated by reduced representation bisulfite sequencing (RRBS) and RNA-Seq. The differentially methylated genes (DMGs) and differentially expressed genes (DEGs) were analyzed by MethylKit R package and edgeR. Correlation analysis between methylation level and mRNA expression was conducted with R software. The functional correlation of DMGs and DEGs was analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, LG tissues from another litter of NOD mice were collected for methylation-specific polymerase chain reaction (MSP) and quantitative real-time PCR (qRT-PCR) to validate the methylation and expression levels of key genes. CD4+ cell infiltration of LGs was detected by immunofluorescence staining. Hypermethylation of LGs was identified in NOD mice with the progression of SS-related dry eye and the DMGs were mainly enriched in the GTPases activation and Ras signaling pathway. RNA-seq analysis revealed 1321, 2549, and 3712 DEGs in the 8-, 12- and 16-week-old NOD mice compared with 4-week-old normal control mice. For GO analysis, the DEGs were mainly enriched in T cell immune responses. Further, a total of 140 MeDEGs were obtained by integrated analysis of methylome and transcriptome, which were primarily enriched in T cell activation, proliferation and differentiation. Based on the main GO terms and KEGG pathways of MeDEGs, 8 genes were screened out. The expression levels of these key genes, especially Itgal, Vav1, Irf4 and Icosl, were verified to elevate after the onset of SS-related dry eye in NOD mice and positively correlated with the extent of inflammatory cell infiltration in LGs. Immunofluorescence assay revealed that CD4+ cell infiltration dramatically increased in LGs of SS-related dry eye mice compared with the control mice. And the expression levels of four genes showed significantly positive correlation with the extent of CD4+ cell infiltration in LGs. MSP showed the hypomethylation of the Irf4 and Itgal promoters in NOD mice with SS-related dry eye compared to control group. Our study revealed the critical role of epigenetic regulation of T cell immunity-related genes in the progression of SS-related dry eye and reminded us that DNA methylation-regulated genes such as Itgal, Vav1, Irf4 and Icosl may be used as new targets for SS-related dry eye therapy.

Reduction of carcinogens in fermented fish (pla-ra and pla-som) by heating.

The risk factors for cholangiocarcinoma (CCA) are opisthorchiasis and the intake of a combination of nitroso compounds through the consumption of traditionally fermented fish, which is very popular in areas where liver flukes are endemic. The incidence of CCA remains high because this cultural habit of rural people has been altered. Therefore, decreasing nitrate and nitrite concentrations in fermented fish are an alternative approach to reducing the risk of CCA. Thus, this study aimed to reduce nitrate and nitrite concentrations in fermented foods by heating and investigated its effect on CCA development in a hamster model. We used Association of Official Analytical Chemists method 973.31 to measure the nitrate and nitrite concentrations in both fermented fish (pla-ra [PR]) and pickled fish (pla-som [PS]) before and after boiling for 5 and 30 min, respectively. The same samples were fed to Opisthorchis viverrini (OV)-infected or -uninfected hamsters for 3 months. Thereafter, the hamsters' liver and blood were collected for analysis. The levels of nitrates and nitrites in PS and PR significantly decreased following boiling for 5 and 30 min. The OV-PR and OV-PS groups showed dramatically increased numbers of inflammatory cells, fibrosis surrounding the bile duct, and focal fibrotic areas. However, after boiling the fermented dishes for 5 and 30 min, the extent of inflammatory cell infiltration and intensity of fibrosis in these groups were decreased. Our findings suggest that boiling reduces nitrate and nitrite toxicity in fermented dishes, as evidenced by reduced hepatic inflammation. However, regardless of heating, kidney tissues are adversely affected when fermented meals are consumed daily.

Diffusion basis spectrum imaging detects pathological alterations in substantia nigra and white matter tracts with early-stage Parkinson's disease.

Using diffusion basis spectrum imaging (DBSI) to examine the microstructural changes in the substantia nigra (SN) and global white matter (WM) tracts of patients with early-stage PD. Thirty-seven age- and sex-matched patients with early-stage PD and 22 healthy controls (HCs) were enrolled in this study. All participants underwent clinical assessments and diffusion-weighted MRI scans, analyzed by diffusion tensor imaging (DTI) and DBSI to assess the pathologies of PD in SN and global WM tracts. The lower DTI fraction anisotropy (FA) was seen in SN of PD patients (PD: 0.316 ± 0.034 vs HCs: 0.331 ± 0.019, p = 0.015). The putative cells marker-DBSI-restricted fraction (PD: 0.132 ± 0.051 vs HCs: 0.105 ± 0.039, p = 0.031) and the edema/extracellular space marker-DBSI non-restricted-fraction (PD: 0.150 ± 0.052 vs HCs: 0.122 ± 0.052, p = 0.020) were both significantly higher and the density of axons/dendrites marker-DBSI fiber-fraction (PD: 0.718 ± 0.073 vs HCs: 0.773 ± 0.071, p = 0.003) was significantly lower in SN of PD patients. DBSI-restricted fraction in SN was negatively correlated with HAMA scores (r = - 0.501, p = 0.005), whereas DTI-FA was not correlated with any clinical scales. In WM tracts, only higher DTI axial diffusivity (AD) among DTI metrics was found in multiple WM regions in PD, while lower DBSI fiber-fraction and higher DBSI non-restricted-fraction were detected in multiple WM regions. DBSI non-restricted-fraction in both left fornix (cres)/stria terminalis (r = -0.472, p = 0.004) and right posterior thalamic radiation (r = - 0.467, p = 0.005) was negatively correlated with MMSE scores. DBSI could potentially detect and quantify the extent of inflammatory cell infiltration, fiber/dendrite loss, and edema in both SN and WM tracts in patients with early-stage PD, a finding remains to be further investigated through more extensive longitudinal DBSI analysis. Our study shows that DBSI indexes can potentially detect early-stage PD's pathological changes, with a notable ability to distinguish between inflammation and edema. This implies that DBSI has the potential to be an imaging biomarker for early PD diagnosis. • Diffusion basis spectrum imaging detected higher restricted-fraction in Parkinson's disease, potentially reflecting inflammatory cell infiltration. • Diffusion basis spectrum imaging detected higher non-restricted-fraction and lower fiber-fraction in Parkinson's disease, indicating the presence of edema and/or dopaminergic neuronal/dendritic loss. • Diffusion basis spectrum imaging metrics correlated with non-motor symptoms, suggesting its potential diagnostic role to detect early-stage PD dysfunctions.

Prevention of Postsurgical Abdominal Adhesion Using Electrospun TPU Nanofibers in Rat Model.

Intra-abdominal adhesions following surgery are a challenging problem in surgical practice. This study fabricated different thermoplastic polyurethane (TPU) nanofibers with different average diameters using the electrospinning method. The conditions were evaluated by scanning electron microscopy (SEM), atomic force microscope (AFM), and Fourier transform infrared spectrometer (FTIR) analysis. A static tensile test was applied using a strength testing device to assess the mechanical properties of the electrospun scaffolds. By changing the effective electrospinning parameters, the best quality of nanofibers could be achieved with the lowest bead numbers. The electrospun nanofibers were evaluated in vivo using a rat cecal abrasion model. The macroscopic evaluation and the microscopic study, including the degree of adhesion and inflammation, were investigated after three and five weeks. The resultant electrospun TPU nanofibers had diameters ranging from about 200 to 1000 nm. The diameters and morphology of the nanofibers were significantly affected by the concentration of polymer. Uniform TPU nanofibers without beads could be prepared by electrospinning through reasonable control of the process concentration. These nanofibers' biodegradability and antibacterial properties were investigated by weight loss measurement and microdilution methods, respectively. The purpose of this study was to provide electrospun nanofibers having biodegradability and antibacterial properties that prevent any adhesions or inflammation after pelvic and abdominal surgeries. The in vivo experiments revealed that electrospun TPU nanofibers reduced the degree of abdominal adhesions. The histopathological study confirmed only a small extent of inflammatory cell infiltration in the 8% and 10% TPU. Conclusively, nanofibers containing 8% TPU significantly decreased the incidence and severity of postsurgical adhesions, and it is expected to be used in clinical applications in the future.

The Extent of Inflammatory Cell Infiltrate and Fibrosis in Lungs of Telomere- and Surfactant-Related Familial Pulmonary Fibrosis.

Familial pulmonary fibrosis (FPF) is a monogenic disease most commonly involving telomere- (TERT) or surfactant- (SFTP) related mutations. These mutations have been shown to alter lymphocytic inflammatory responses, and FPF biopsies with histological lymphocytic infiltrates have been reported. Recently, a model of a surfactant mutation in mice showed that the disease initially started with an inflammatory response followed by fibrogenesis. Since inflammation and fibrogenesis are targeted by different drugs, we investigated whether the degree of these two features co-localize or occur independently in different entities of FPF, and whether they influence survival. We quantified the number of lymphocyte aggregates per surface area, the extent of diffuse lymphocyte cell infiltrate, the number of fibroblast foci per surface area, and the percentage of fibrotic lung surface area in digitally scanned hematoxylin and eosin (H&E) sections of diagnostic surgical biopsies of patients with TERT-related FPF (TERT-PF; n = 17), SFTP-related FPF (SFTP-PF; n = 7), and sporadic idiopathic pulmonary fibrosis (sIPF; n = 10). For comparison, we included biopsies of patients with cellular non-specific interstitial pneumonia (cNSIP; n = 10), an inflammatory interstitial lung disease with high lymphocyte influx and usually responsive to immunosuppressive therapy. The degree of inflammatory cell infiltrate and fibrosis in TERT-PF and SFTP-PF was not significantly different from that in sIPF. In comparison with cNSIP, the extent of lymphocyte infiltrates was significantly lower in sIPF and TERT-PF, but not in SFTP-PF. However, in contrast with cNSIP, in sIPF, TERT-PF, and SFTP-PF, diffuse lymphocyte cell infiltrates were predominantly present and lymphocyte aggregates were only present in fibrotic areas (p < 0.0001). Furthermore, fibroblast foci and percentage of fibrotic lung surface were associated with survival (p = 0.022 and p = 0.018, respectively), while this association was not observed for lymphocyte aggregates or diffuse lymphocytic infiltration. Inflammatory cells in diagnostic lung biopsies of TERT-PF, SFTP-PF, and sIPF were largely confined to fibrotic areas. However, based on inflammation and fibrosis, no differences were found between FPF and sIPF, substantiating the histological similarities between monogenic familial and sporadic disease. Furthermore, the degree of fibrosis, rather than inflammation, correlates with survival, supporting that fibrogenesis is the key feature for therapeutic targeting of FPF.

CD45 Immunohistochemistry in Mouse Kidney.

CD45 is a pan-leukocyte marker, and CD45 stain is widely used to determine the extent of inflammatory cell infiltration and its association with tissue injury. In this manuscript, we share a reliable immunohistochemistry (IHC) protocol for CD45 staining in sections of paraffin-embedded mouse kidney. A rat anti-CD45 antibody was used as primary antibody, and a mouse adsorbed biotin-conjugated goat anti-rat IgG was selected as secondary antibody. A horseradish peroxidase (HRP)-linked avidin/biotin detection system was used to amplify the signal, which was detected with 3,3'-Diaminobenzidine (DAB). With this protocol, we show that the CD45 antibody recognizes cells of hematolymphoid lineage in bone marrow, as well as monocyte/macrophages in liver and lung tissue. The utility of this protocol in pathology research was indicated by dramatically increased CD45-positive (CD45+) cells in the kidneys of a mouse model of diabetes. Double staining for CD45 and injury marker KIM-1 showed accumulated CD45+ cells around injured tubular cells. CD45 and F4/80 macrophage staining on adjacent tissue sections revealed overlap of CD45+ cells with other inflammatory cells.

Study on Prevention of Postoperative Abdominal Adhesions with PLGA Nanofiber Membrane

Objective: To evaluate the effectiveness of poly (lactic-co-glycolic acid) (PLGA) nanofiber membrane on prevention of postoperative abdominal adhesion. Methods: PLGA nanofiber membrane was prepared by high-voltage electrospinning technique. The effectiveness of the membrane in prevention of postoperative abdominal adhesions was characterized with rat abdominal adhesion models. Results: PLGA nanofiber membrane was prepared successfully by electrospinning technique. Scanning electron microscopy (SEM) observation showed that the average diameter of PLGA fibers was approximately 800 nm, and the membrane had microporous structures. Mechanical tests showed that the tensile strength of PLGA nanofiber membrane was 6.36 ± 0.39 MPa, which was significantly higher than the tensile strength of DIKANG absorbable medical film. The results of in vivo experiments showed that PLGA nanofiber membrane and DIKANG absorbable medical film could both reduce the degree of abdominal adhesions. The histological results showed that there was only a small extent of inflammatory cell infiltration in the PLGA group and the control group. The proliferation of connective tissue was reduced, and so was the degree of adhesion. Conclusion: PLGA nanofiber membrane can significantly reduce the incidence and severity of postoperative adhesions, and bodes well for future clinical applications.

Cardioprotective effects of garcinol following myocardial infarction in rats with isoproterenol-induced heart failure

Myocardial infarction is a clinical form of necrosis in the myocardium caused by an imbalance between the coronary blood supply and myocardial demand. Garcinol is a polyisoprenylated benzophenone found in the fruit of Garcinia indica, which is abundant in tropical regions. This fruit contains high levels of garcinol, isoxanthochymol, isogarcinol, hydroxycitric acid and xanthochymol. Garcinol and hydroxycitric acid have been shown to have antioxidant effects. In this study, rats were assigned to sham, control, low-dose, high-dose and positive control groups. Hemodynamic and apoptotic markers were evaluated, and histopathological analysis was conducted. The mRNA and protein levels of caspase-3, Bax, Bcl-2 and cleaved caspase-3 were quantified. Garcinol treatment increased the heart rate and improved the maximum rate of increase in left-ventricle (LV) pressure (+dp/dtmax), maximum rate of decrease in LV pressure (–dp/dtmax), LV ejection fraction and LV systolic pressure in rats with induced heart failure. Garcinol treatment reversed body, liver and heart weight changes, resulting in returns to near-normal levels. In the garcinol treatment group, the number of broken fibers, extent of inflammatory cell infiltration and rate of apoptosis remained within normal ranges. Garcinol reduced the cross-sectional areas of cardiomyocytes, and reduced interstitial fibrosis to a normal level. The mRNA and protein levels of cleaved caspase-3, caspase-3 and Bax were reduced, whereas those of Bcl-2 were increased, following high-dose (100 mg/kg) garcinol treatment. These findings suggest that garcinol effectively prevents apoptosis in rats with isoproterenol-induced heart failure and in cardiac H9C2 cells.

Oligodendrocytes that survive acute coronavirus infection induce prolonged inflammatory responses in the CNS

Neurotropic strains of mouse hepatitis virus (MHV), a coronavirus, cause acute and chronic demyelinating encephalomyelitis with similarities to the human disease multiple sclerosis. Here, using a lineage-tracking system, we show that some cells, primarily oligodendrocytes (OLs) and oligodendrocyte precursor cells (OPCs), survive the acute MHV infection, are associated with regions of demyelination, and persist in the central nervous system (CNS) for at least 150 d. These surviving OLs express major histocompatibility complex (MHC) class I and other genes associated with an inflammatory response. Notably, the extent of inflammatory cell infiltration was variable, dependent on anatomic location within the CNS, and without obvious correlation with numbers of surviving cells. We detected more demyelination in regions with larger numbers of T cells and microglia/macrophages compared to those with fewer infiltrating cells. Conversely, in regions with less inflammation, these previously infected OLs more rapidly extended processes, consistent with normal myelinating function. Together, these results show that OLs are inducers as well as targets of the host immune response and demonstrate how a CNS infection, even after resolution, can induce prolonged inflammatory changes with CNS region-dependent impairment in remyelination.

Muscle Weakness in Myositis: MicroRNA-Mediated Dystrophin Reduction in a Myositis Mouse Model and Human Muscle Biopsies.

ObjectiveMuscle inflammation is a feature in myositis and Duchenne muscular dystrophy (DMD). Autoimmune mechanisms are thought to contribute to muscle weakness in patients with myositis. However, a lack of correlation between the extent of inflammatory cell infiltration and muscle weakness indicates that nonimmune pathologic mechanisms may play a role. The present study focused on 2 microRNA (miRNA) sets previously identified as being elevated in the muscle of patients with DMD—an “inflammatory” miRNA set that is dampened with glucocorticoids, and a “dystrophin‐targeting” miRNA set that inhibits dystrophin translation—to test the hypothesis that these miRNAs are similarly dysregulated in the muscle of patients with myositis, and could contribute to muscle weakness and disease severity.MethodsA major histocompatibility complex class I–transgenic mouse model of myositis was utilized to study gene and miRNA expression and histologic features in the muscle tissue, with the findings validated in human muscle biopsy tissue from 6 patients with myositis. Mice were classified as having mild or severe myositis based on transgene expression, body weight, histologic disease severity, and muscle strength/weakness.ResultsIn mice with severe myositis, muscle tissue showed mononuclear cell infiltration along with elevated expression of type I interferon and NF‐κB–regulated genes, including Tlr7 (3.8‐fold increase, P < 0.05). Furthermore, mice with severe myositis showed elevated expression of inflammatory miRNAs (miR‐146a, miR‐142‐3p, miR‐142‐5p, miR‐455‐3p, and miR‐455‐5p; ~3–40‐fold increase, P < 0.05) and dystrophin‐targeting miRNAs (miR‐146a, miR‐146b, miR‐31, and miR‐223; ~3–38‐fold increase, P < 0.05). Bioinformatics analyses of chromatin immunoprecipitation sequencing (ChIP‐seq) data identified at least one NF‐κB consensus element within the promoter/enhancer regions of these miRNAs. Western blotting and immunofluorescence analyses of the muscle tissue from mice with severe myositis demonstrated reduced levels of dystrophin. In addition, elevated levels of NF‐κB–regulated genes, TLR7, and miRNAs along with reduced dystrophin levels were observed in muscle biopsy tissue from patients with histologically severe myositis.ConclusionThese data demonstrate that an acquired dystrophin deficiency may occur through NF‐κB–regulated miRNAs in myositis, thereby suggesting a unifying theme in which muscle injury, inflammation, and weakness are perpetuated both in myositis and in DMD.

Blood urea nitrogen may serve as a predictive indicator of retained placenta in dairy cows

Retained placentae (RP) results in significant economic losses to dairy farmers. In Experiment 1, to screen biochemical indicators of RP, 21 cows with RP and 21 cows with no retained placenta (NRP) were selected as a control group, and blood was collected at -7 d, 0 h (parturition) and 12 h. Serum biochemical indicators were ascertained. Results indicate serum concentrations of phosphorus (P) and blood urea nitrogen (BUN) in cows of the RP group were markedly greater than in cows of the NRP group at -7 d (P < 0.01). In Experiment 2, to evaluate predictive indicators for RP, 34 cows with RP and 34 cows with NRP were selected, and there was blood sampling at -15 d, -10 d, -7 d, -4 d, and -1 d. Serum P, BUN, and total protein (TP) were evaluated. Associations of values among the three indicators and occurrence of RP were analyzed using binary logistic regression. Results indicate there was a negative correlation between only the values for BUN and RP (P = 0.016). In Experiment 3, to test hypothesis that relatively greater concentrations of BUN effects immune function in placental tissues, four cows were selected, placentae were collected at 0 and 12 h, and hematoxylin-eosin (HE) staining was performed. Results indicated that the extent of inflammatory cell infiltration and vascular proliferation were less at the 12 than 0-hour timepoint. Taken together, BUN at -7 d may serve as a predictive indicator of RP in cows.

Effects of particulate matter from straw burning on lung fibrosis in mice

ObjectiveTo investigate the impacts of particulate matter 2.5 (PM2.5) from straw burning on the acute exacerbation of lung fibrosis in mice and the preventive effects of N-acetylcysteine (NAC). MethodsThe composition, particle size, and 30-min concentration change in an exposure system of the PM2.5 from straw-burning were determined. Forty C57BL male mice were equally randomized to two groups: bleomycin (BLM)-induced lung fibrosis with an exposure to air (BLM+air) and BLM+PM2.5 groups. On day 7 after receiving intratracheal injection of BLM, mice were exposed to air or PM2.5 in an exposure system for 30min twice daily and then sacrificed after one-week or four-week exposure (10 mice/group). Mouse survival, lung histopathology, macrophage accumulation in the lung, and pro-inflammatory cytokine levels in alveolar lavage fluid (ALF) were determined. ResultsPM2.5 from straw burning were mainly composed of organic matter (74.1%); 10.92% of the inorganic matter of the PM2.5 were chloride ion; 4.64% were potassium ion; other components were sulfate, nitrate, and nitrite. Particle size was 10nm–2μm. Histopathology revealed a greater extent of inflammatory cell infiltration in the lung, widened alveolar septum, and lung fibrosis in the BLM+PM2.5 group than in the BLM+air group and a greater extent of those adverse effects after four-week than after one-week exposure to PM2.5. The BLM+PM2.5 group also showed macrophages containing particular matter and increased pulmonary collagen deposition as the exposure to PM2.5 increased. Interleukin (IL)-6 and TNF-α levels in ALF were significantly higher in the BLM+PM2.5 group than in the BLM+air group (P<0.05) and significantly higher after four-week exposure than after one-week exposure to PM2.5 (P<0.05). TGF-β levels in ALF after four-week exposure were significantly higher in the BLM+PM2.5 group than in the BLM+air group (P<0.05). The levels of IL-6, TNF-α, and TGF-β in peripheral serum were not significantly different in the BLM+PM2.5 and BLM+air groups. Lung hydroxyproline contents increased as the exposure to PM2.5 increased and were significantly higher after four-week than after one-week exposure (P=0.019). Exposure to PM2.5 did not affect the survival of normal mice (100%) but reduced the survival of mice with BLM-induced IPF (30%), whereas NAC extended the survival (70%, vs. BLM+PM2.5, P=0.032). ConclusionExposure of mice with BLM-induced IPF to PM2.5 from straw burning exacerbated lung inflammation and fibrosis and increased mortality; NAC increased the mouse survival, indicating protective effects.

Pharmacological Effect of Caulophyllum robustum on Collagen-Induced Arthritis and Regulation of Nitric Oxide, NF-κB, and Proinflammatory Cytokines In Vivo and In Vitro.

Caulophyllum robustum Maxim (C. robustum) has commonly been used as traditional Chinese medicine for the treatment of rheumatic pain and rheumatoid arthritis (RA) in China. This paper first investigated the anti-inflammation effect of C. robustum extraction (CRME) on RAW264.7 cells stimulated by lipopolysaccharide (LPS) and gene expression levels of inflammatory factors. Moreover, we first evaluated the anti-RA effects of CRME using collagen-induced arthritis (CIA) in DBA/1J mice, and the incidence, clinical score, and joint histopathology were evaluated. The levels of IL-1, IL-6, TNF-α, and PGE2 inflammatory factors in sera of mice were detected by enzyme-linked immunosorbent assay. The expression of NF-κB p65 in the joint was tested by immune histochemical technique. The results showed that, compared with the model group, CRME significantly improved symptoms of the arthritis index, limb swelling, and histological findings by decreasing synovial membrane damage, the extent of inflammatory cell infiltration, and the expansion of capillaries in CIA mice. The results also showed that CRME can reduce the levels of IL-1, IL-6, TNF-α, and PGE2 and inhibit the expression of NF-κB p65. All these results indicated the anti-inflammatory efficacy of CRME as a novel botanical extraction for the treatment of RA.

Effect of sphingosine kinase modulators on interleukin-1β release, sphingosine 1-phosphate receptor 1 expression and experimental autoimmune encephalomyelitis.

The sphingosine analogue, FTY720 (GilenyaR ), alleviates clinical disease progression in multiple sclerosis. Here, we variously assessed the effects of an azide analogue of (S)-FTY720 vinylphosphonate (compound 5; a sphingosine kinase 1 activator), (R)-FTY720 methyl ether (ROMe, a sphingosine kinase 2 inhibitor) and RB-020 (a sphingosine kinase 1 inhibitor and sphingosine kinase 2 substrate) on IL-1β formation, sphingosine 1-phosphate levels and expression of S1P1 receptors. We also assessed the effect of compound 5 and ROMe in an experimental autoimmune encephalomyelitis (EAE) model in mice. We measured IL-1β formation by macrophages, sphingosine 1-phosphate levels and expression levels of S1P1 receptors in vitro and clinical score in mice with EAE and the extent of inflammatory cell infiltration into the spinal cord in vivo. Treatment of differentiated U937 macrophages with compound 5, RB-020 or sphingosine (but not ROMe) enhanced IL-1β release. These data suggest that these compounds might be pro-inflammatory in vitro. However, compound 5 or ROMe reduced disease progression and infiltration of inflammatory cells into the spinal cord in EAE, and ROMe induced a reduction in CD4+ and CD8+ T-cell levels in the blood (lymphopenia). Indeed, ROMe induced a marked decrease in expression of cell surface S1P1 receptors in vitro. This is the first demonstration that an activator of sphingosine kinase 1 (compound 5) and an inhibitor of sphingosine kinase 2 (ROMe, which also reduces cell surface S1P1 receptor expression) have an anti-inflammatory action in EAE.

Inflammatory cell infiltrates in the heart of patients with coronary artery disease with and without inflammatory rheumatic disease: a biopsy study.

BackgroundThe cause of premature cardiovascular disease (CVD) in inflammatory rheumatic diseases (IRDs) has not been fully elucidated. As inflammation may play a role, we wanted to compare the occurrence and extent of inflammatory cell infiltrates (ICIs), small vessel vasculitis, and the amount of adipose tissue and collagen in cardiac biopsies taken from patients with coronary artery disease with and without IRDs.MethodsFrom among the Feiring Heart Biopsy Study subjects, we selected patients undergoing coronary artery bypass grafting from whom paraffin-embedded, formalin-fixed specimens from the right atrium were available. The sample comprised 48 patients with IRD and 40 non-IRD patients. Hematoxylin and eosin staining was used to examine the presence and location of ICIs and vasculitis, and Lendrum (Martius yellow, scarlet, and blue) staining was carried out for collagen and adipose tissue.ResultsEpicardial ICIs were found in 27 (56 %) patients with IRD and 24 (60 %) non-IRD patients. There were no significant differences between patients with IRD and non-IRD patients in the amount of cardiac ICIs and adipose tissue, but patients with IRD had more collagen in the myocardium than non-IRD patients. Small vessel vasculitis was not observed in any cardiac specimen. Patients with epicardial ICIs were, on average, 7 years younger than those without.ConclusionsOur results do not support the notion that inflammation in cardiac peri-, epi-, and myocardium plays a more important role in CVD of patients with IRD than non-IRD patients. The increased amount of collagen in the myocardium of patients with IRD suggests differences in extracellular matrix composition and/or mass, which might play a role in cardiac remodeling, and represent targets for novel therapies against heart failure.

BMP Antagonist Gremlin 2 Limits Inflammation After Myocardial Infarction.

We have recently shown that the bone morphogenetic protein (BMP) antagonist Gremlin 2 (Grem2) is required for early cardiac development and cardiomyocyte differentiation. Our initial studies discovered that Grem2 is strongly induced in the adult heart after experimental myocardial infarction (MI). However, the function of Grem2 and BMP-signaling inhibitors after cardiac injury is currently unknown. To investigate the role of Grem2 during cardiac repair and assess its potential to improve ventricular function after injury. Our data show that Grem2 is transiently induced after MI in peri-infarct area cardiomyocytes during the inflammatory phase of cardiac tissue repair. By engineering loss- (Grem2(-/-)) and gain- (TG(Grem2)) of-Grem2-function mice, we discovered that Grem2 controls the magnitude of the inflammatory response and limits infiltration of inflammatory cells in peri-infarct ventricular tissue, improving cardiac function. Excessive inflammation in Grem2(-/-) mice after MI was because of overactivation of canonical BMP signaling, as proven by the rescue of the inflammatory phenotype through administration of the canonical BMP inhibitor, DMH1. Furthermore, intraperitoneal administration of Grem2 protein in wild-type mice was sufficient to reduce inflammation after MI. Cellular analyses showed that BMP2 acts with TNFα to induce expression of proinflammatory proteins in endothelial cells and promote adhesion of leukocytes, whereas Grem2 specifically inhibits the BMP2 effect. Our results indicate that Grem2 provides a molecular barrier that controls the magnitude and extent of inflammatory cell infiltration by suppressing canonical BMP signaling, thereby providing a novel mechanism for limiting the adverse effects of excessive inflammation after MI.

Establishment of rat model of central venous catheter (CVC): associated infection and evaluation of the virulence of bacterial biofilms.

In this study, a central venous catheter (CVC)-associated infection model was established in rats to investigate and evaluate the effect of biofilms on the virulence of the pathogens. Twenty-four adult SD rats were randomly divided into biofilm positive (BF+) and biofilm negative (BF-) groups to be challenged with strains of S.epidermidis. Serum levels of inflammatory cytokines were measured and the infection rate and counts of bacteria cells were studied. Compared to rats of BF- group, the serum level of TNF and IL-6 significantly increased in rats of BF+ group (P < 0.01) and the level of IL-10 and IFN-γ significantly decreased (P < 0.01), striking the balance of pro-inflammatory/anti-inflammatory cytokines. The infection rate and bacterial counts in tissues and blood of rats of BF + group were significantly higher than those of rats of BF- group (P < 0.05).Inflammatory cell infiltration in vital organs (heart, lung, liver and kidneys) was more significant in rats of BF+ group than that of rats of BF- group. CVC-associated infection model can be successfully reproduced in rats by injecting 5 × 10(6) CFU of S.epidermidis. Biofilm formation can significantly enhance the virulence of the bacteria, leading to uncontrolled infection. The serum level of inflammatory cytokines, infection rate and the extent of inflammatory cell infiltration are important markers for evaluating the virulence of biofilm.

Transient filiform papillitis in a child.

Dear Editor: The tongue is covered with many papillae, and these papillae primarily consist of the fungiform and filiform types. Histologically, filiform papillae show finger-like projections, and they have a core of loose connective tissue covered with keratinized stratified squamous epithelium, whereas fungiform papillae are round and covered with lightly or non-keratinized stratified squamous epithelium1. Transient lingual papillitis (TLP) is an inflammatory disorder that has a clinically abrupt onset of pain, rapid disappearance of papules on the tongue, and inflammatory cell infiltration with some ulcerations on the fungiform papillae that is observed on histopathological examination2. Herein, we describe a child showing clinical TLP features but the histopathological findings indicated primary involvement of filiform rather than fungiform papillae. An 8-year-old Korean healthy boy presented with a 1-day history of asymptomatic papules of the tongue. Physical examination revealed diffuse erythematous papules prominently observed on the lingual papillae on the dorsal surface of the tongue (Fig. 1). Biopsy showed tapered finger-like projections covered with keratinized stratified squamous epithelium, suggesting the involvement of filiform rather than fungiform papillae. Acanthosis, papillomatosis, and parakeratosis with sparse lymphocytic inflammatory cell infiltration in the lamina propria were noted (Fig. 2). To rule out herpes simplex virus (HSV) infection, polymerase chain reaction (PCR) was performed for amplification of a subunit of the deoxyribonucleic acid polymerase gene, by using an HSV Type 1/2 PCR kit (Bio-Core Inc., Seoul, Korea); negative results were obtained. After 14 days, the patient's parents reported that the lingual papules had cleared after a few days without any sequelae. From that time, the patient has occasionally developed papules on the tongue followed by spontaneous improvement. The final diagnosis was difficult to make because although his clinical progression was similar to asymptomatic TLP, the histological findings primarily involved filiform papillae, which exhibited a lesser extent of inflammation in contrast to the inflammatory and ulcerated changes that are associated with fungiform papillae in TLP. TLP progresses transiently because the pain subsides within a few days, and it is referred to as 'lie bumps' owing to its characteristics2,3. Recently, some authors reviewed 9 patients who showed long-duration adult-onset papillitis and termed this condition 'chronic lingual papulosis (CLP)'4. CLP represents multiple asymptomatic but enlarged lingual papules that microscopically exhibit dense, fibrous, hyperplastic filiform papillae. The present case was distinct from CLP in that the age of onset was much younger, the clinical course was transient, and our patient had no underlying lingual or oral abnormalities. It also showed some features of TLP and some characteristics of CLP. Although abrupt onset and rapid disappearance of tongue papules are clinical features of TLP, the signs of filiform papillae involvement with less extent of inflammatory cell infiltration observed on histological examination were indicative of CLP. In the present report, we introduce an interesting case of lingual papules characterized by an abrupt onset followed by the rapid disappearance of lingual papulosis with microscopic features of filiform papillitis. We believe that this case can be proposed as a new disease entity, specifically a filiform-involving lingual disorder, if similar cases are subsequently reported. Fig. 1 Multiple erythematous papules on the dorsal surface of the tongue. Fig. 2 Finger-like projecting structures covered with keratinized squamous epithelium exhibiting filiform papillae and sparse lymphocytic inflammatory cell infiltration in the lamina propria (H&E, ×100).

Differential role of MyD88 and Mal/TIRAP in TLR2-mediated gastric tumourigenesis

Signalling by the toll-like receptor (TLR) family of pathogen recognition receptors has emerged as a key molecular component in the pathogenesis of an increasing number of inflammatory-related cancers, among which gastric cancer rates as the second most lethal cancer world-wide. The myeloid differentiation factor 88 (MyD88) adapter molecule has a critical role in mediating innate immune signalling by members of the TLR and interleukin (IL)-1 families, and has been associated with either pro- or antitumourigenic responses in various cancer models. However, little is known about the in vivo role of MyD88 adapter-like (Mal)/TIR-domain containing adapter protein (TIRAP), which is restricted to facilitating TLR4 and TLR2 signalling. To interrogate the role of these innate immune signalling components in gastric tumourigenesis, here we have employed the spontaneous gastric cancer gp130(F/F) mouse model, in which TLR2 promotes the growth of gastric tumours. Genetic ablation of Myd88 in gp130(F/F) mice suppressed tumourigenesis and was associated with increased apoptosis and reduced proliferation in the gastric tumour epithelium, comparable to that observed previously upon deletion of Tlr2 in gp130(F/F) mice. By contrast, the tumour burden in gp130(F/F):Mal(-/-) mice was equivalent to their gp130(F/F) littermates. At the molecular level, suppressed tumourigenesis in gp130(F/F):Myd88(-/-) mice correlated with reduced expression and activation of TLR2-regulated protumourigenic genes and signalling pathways, respectively. Consistent with the previously defined non-essential role for TLR2 in gastric tumour inflammation, the extent of inflammatory cell infiltrates in gastric tumours from gp130(F/F):Mal(-/-) and gp130(F/F):Myd88(-/-) mice remained unaltered compared with gp130(F/F) mice. Collectively, our data reveal a differential, but inflammation-independent, requirement for Mal and MyD88 during TLR2-promoted gastric tumourigenesis.