Evodiamine Relieve LPS-Induced Mastitis by Inhibiting AKT/NF-κB p65 and MAPK Signaling Pathways.

Evodiamine, an alkaloid component in the fruit of Evodia, has been shown to have biological functions such as antioxidant and anti-inflammatory. But whether evodiamine plays an improvement role on mastitis has not been studied. To investigate the effect and mechanism of evodiamine on lipopolysaccharide (LPS)-induced mastitis was the purpose of this study. In animal experiments, the mouse mastitis model was established by injecting LPS into the canals of the mammary gland. The results showed that evodiamine could significantly relieve the pathological injury of breast tissue and the production of pro-inflammatory cytokines and inhibit the activation of inflammation-related pathways such as AKT, NF-κB p65, ERK1/2, p38, and JNK. In cell experiments, the mouse mammary epithelial cells (mMECs) were incubated with evodiamine for 1 h and then stimulated with LPS. Next, pro-inflammatory mediators and inflammation-related signal pathways were detected. As expected, our results showed that evodiamine notably ameliorated the inflammatory reaction and inhibit the activation of related signaling pathways of mMECs. All the results suggested that evodiamine inhibited inflammation by inhibiting the phosphorylation of AKT, NF-κBp65, ERK1/2, p38, and JNK thus the LPS-induced mastitis was ameliorated. These findings suggest that evodiamine maybe a potential drug for mastitis because of its anti-inflammatory effects.

Effect of Palrnatine on lipopolysaccharide-induced acute lung injury by inhibiting activation of the Akt/NF|-κB pathway.

Inflammation plays an important role in the development of acute lung injury (ALI). Severe pulmonary inflammation can cause acute respiratory distress syndrome (ARDS) or even death. Expression of proinflammatory interleukin-|1ß (IL-|1ß) and inducible nitric oxide synthase (iNOS) in the process of pulmonary inflammation will further exacerbate the severity of ALI. The purpose of this study was to explore the effect of Palrnatine (Pa) on lipopolysaccharide (LPS)-induced mouse ALI and its underlying mechanism. Pa, a natural product, has a wide range of pharmacological activities with the potential to protect against lung injury. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to detect the expression and translation of inflammatory genes and proteins in vitro and in vivo. Immunoprecipitation was used to detect the degree of P65 translocation into the nucleus. We also used molecular modeling to further clarify the mechanism of action. The results showed that Pa pretreatment could significantly inhibit the expression and secretion of the inflammatory cytokine IL-1ß, and significantly reduce the protein level of the proinflammatory protease iNOS, in both in vivo and in vitro models induced by LPS. Further mechanism studies showed that Pa could significantly inhibit the activation of the protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway in the LPS-induced ALI mode and in LPS-induced RAW264.7 cells. Through molecular dynamics simulation, we observed that Pa was bound to the catalytic pocket of Akt and effectively inhibited the biological activity of Akt. These results indicated that Pa significantly relieves LPS-induced ALI by activating the Akt/NF-κB signaling pathway.

Pedunculoside protects against LPS-induced mastitis in mice by inhibiting inflammation and maintaining the integrity of blood-milk barrier.

Mastitis is a disease that seriously threatens the health of the mammary gland after delivery. Pedunculoside (PE) is the main bioactive component of Aquifoliaceae. The purpose of this experiment is to explore the effects of PE on mastitis and its underlying mechanisms. Our research results showed that PE could significantly inhibit the increase in the levels of inflammatory mediators such as TNF-α, IL-6, IL-1ß, MPO and iNOS during mastitis. Mechanism studies have found that PE could significantly inhibit the phosphorylation of AKT protein and binds to the ASP-184 site. Further research found that PE also inhibited the activation of AKT's downstream pro-inflammatory signals NF-κB and MAPK. In addition, PE effectively promote the expression of tight junction proteins occludin and claudin-3 during inflammation, maintaining the integrity of the blood-milk barrier. In summary, our research shows that PE inhibits the phosphorylation of AKT/NF-κB and MAPK signals; It also relieves mastitis by repairing the blood-milk barrier.

Antilingula as a Surgical Reference Point for Vertical Ramus Osteotomy.

PURPOSE: This study investigated the antilingula and its related landmarks, the mandibular rami, by using cone-beam computed tomography (CBCT). METHODS: CBCT images of 37 patients (74 sides of the mandibular ramus) were collected. The landmarks of antilingula (AntiL), anterior ramus (A), posterior ramus (P), superior ramus (S), and inferior ramus (I) were identified. The distances (A-AntiL, P-AntiL, S-AntiL, and I-AntiL) were statistically evaluated according to gender, side (right and left), and skeletal patterns. RESULTS: The distance from the antilingula to the anterior (A-AntiL) border of the ramus was significantly longer on the right side (14.69 mm) than on the left side (13.97 mm). Male patients had longer AntiL-P, AntiL-I, and S-I distances (18.96, 40.07, and 54.94 mm, respectively) than did female patients (16.66, 35, and 47.54 mm, respectively). Regarding skeletal patterns, the classes can be ordered as follows in terms of the measurements: class III>class II>class I. However, the differences between the classes were nonsignificant. Pearson correlation analysis revealed that gender and S-I distance were strongly correlated (r = 0.667); specifically, male patients had a longer S-I distance. A-AntiL and A-P also exhibited a strong correlation (r = 0.796). CONCLUSION: Antilingula-related distances did not differ between skeletal patterns. Among antilingula-related variables, A-AntiL could serve as a favorable measuring point during operation.

Importance in the Occurrence Distribution of Minimum Oropharyngeal Cross-Sectional Area in the Different Skeletal Patterns Using Cone-Beam Computed Tomography.

PURPOSE: Obstructive sleep apnea is a condition involving repetitive partial or complete collapse of the pharyngeal airway, especially in patient with mandibular hypoplasia. The present study investigated the differences between the volume of the oropharyngeal airway and the minimum axial area in three skeletal patterns through the use of cone-beam computed tomography (CBCT). MATERIALS AND METHODS: CBCT scans of 147 patients were collected to measure the upper oropharyngeal airway volume (UOV), lower oropharyngeal airway volume (LOV), upper oropharyngeal airway area (UOA), minimum upper oropharyngeal airway area (MUOA), lower oropharyngeal airway area (LOA), minimum lower oropharyngeal airway area (MLOA), anatomical structures (orbitale, Or; porion, Po; pogonion, Pog; hyoid, H; second cervical vertebra, C2; fourth cervical vertebra, C4), and relevant angles. Statistical analysis was performed using analysis of variance and Pearson's test. RESULTS: Compared with patients in Class II, those in Class III and Class I exhibited a significantly anterior position of H and Pog. The vertical positions of H and Pog revealed no significant difference between the three skeletal patterns. Patients in skeletal Class III exhibited significantly larger oropharyngeal area (UOA, MUOA, LOA, MLOA) and oropharyngeal airway (UOV and LOV) than those in skeletal Class II did. The horizontal position of Pog had a moderately significant correlation with UOA (r = 0.471) and MUOA (r = 0.455). CONCLUSION: Patients in skeletal Class II had significantly smaller oropharyngeal airway areas and volumes than those in Class III did. The minimum oropharyngeal cross-sectional area had a 67% probability of occurrence in the upper oropharyngeal airway among patients in Class I and Class II and a 50% probability of occurrence among patients in Class III.

Protective effect of myricetin on LPS-induced mastitis in mice through ERK1/2 and p38 protein author.

The inflammatory reaction of mammary gland tissue in dairy cattle leads to the occurrence of mastitis disease and causes huge economic loss. Myricetin (Myr), a flavonoid natural product, is extracted from the root, stem, and leaves of Myrica rubra. It has a wide range of biological activities, such as anti-oxidant, anti-inflammatory, and anti-tumor. The purpose of this experiment is to further explore the effect of Myr on mastitis and further explore its potential mechanism in LPS-induced mice mastitis model and LPS-induced mice mammary epithelial cells (mMECs). The results showed that Myr could significantly inhibit the expression of TNF-α, IL-6, and IL-1ß in the mammary gland of mice. Furthermore, the results of mechanism studies show that Myr can significantly inhibit P38 and ERK1/2 protein phosphorylation levels in mice mammary tissue, and this result has been further verified at the cellular level. These results confirm that Myr can significantly inhibit mammary inflammation, and its potential mechanism is to play a protective role by inhibiting the phosphorylation level of P38 and ERK1/2 protein.

Myricetin protects against H2 O2 -induced oxidative damage and apoptosis in bovine mammary epithelial cells.

High-producing dairy cows are prone to oxidative stress due to their high secretion and strong metabolism, and excessive oxidative stress may cause the apoptosis of bovine mammary epithelial cells (bMECs). Myricetin (Myr) has been shown to have a wide range of pharmaceutical activities. The aim of this study was to evaluate the effect of Myr on hydrogen peroxide (H2 O2 )-induced oxidative stress and apoptosis in bMECs and to clarify the underlying mechanism. bMECs were pretreated with or without Myr and then stimulated with H2 O2 . The results showed that Myr significantly increased the total antioxidant capacity and superoxide dismutase levels and decreased the malondialdehyde (MDA) and reactive oxygen species (ROS) levels in a model of oxidative stress induced by H2 O2 in bMECs. Mechanistic studies found that Myr inhibited H2 O2 -induced oxidative stress in bMECs through the adenosine monophosphate-activated protein kinase/nuclear factor erythroid-2 related factor 2 (AMPK/NRF2) signaling pathway. Additional research found that Myr could also inhibit H2 O2 -induced apoptosis in bMECs through NRF2. These data suggest that Myr effectively alleviated oxidative stress and apoptosis in H2 O2 -induced bMECs through the activation of the AMPK/NRF2 signaling pathway.

The correlation between surgical reference points: antilingula, lingula, and mandibular foramen.

The purpose of the present study was to investigate and determine the anatomical relationship between the antilingula, lingula, and mandibular foramen using cone-beam computed tomography (CBCT). METHODS: CBCT images of 90 participants (180 mandibular ramus) were collected. The locations of and distances between the antilingula, lingual, and mandibular foramen according to side (right and left) and skeletal patterns were measured and then evaluated by statistical analysis. RESULTS: Only 27 participants (15%) had bilateral distinct antilingula, lingula, and mandibular foramen. The antilingula was located anteriorly (4.28 mm and 3.59 mm) and above (1.99 mm and 8.52 mm) the lingula and mandibular foramen. The lingula was behind (0.69 mm) and above (6.53 mm) the mandibular foramen. Skeletal Class III was anterior and inferior to Class II and Class I in the antilingula, lingula, and mandibular foramen. Considering the correlations of landmarks, we found that the lingula was strongly correlated with the mandibular foramen on the X axis (r = 0.757) and Y-axis (r = 0.878). CONCLUSION: The antilingula is located anteriorly and above the lingula and mandibular foramen. The lingula is behind and above the mandibular foramen. The osteotomy line of orthognathic surgery can only be safely designed through actual measurement of the locations of the antilingula, lingula, and mandibular foramen.

Colorectal Cancer Diagnostic Algorithm Based on Sub-Patch Weight Color Histogram in Combination of Improved Least Squares Support Vector Machine for Pathological Image.

In order to improve the diagnostic accuracy of colon cancer, a novel classification algorithm based on sub-patch weight color histogram and improved SVM is proposed, which has good approximation ability for complex pathological image. Our proposed algorithm combines wavelet kernel SVM with color histogram to classify pathological image. Firstly, the pathological image is divided into non-overlapping sub-patches, and the features of sub-patch histogram are extracted. Then, the global and local features are fused by the sub-patch weighting algorithm. Then, the RelicfF based forward selection algorithm is used to integrate color features and texture features so as to enhance the characterization capabilities of the tumor cell. Finally, Morlet wavelet kernel-based least squares support vector machine method is adopted to enhance the generalization ability of the model for small sample with non-linear and high-dimensional pattern classification problems. Experimental results show that the proposed pathological diagnostic algorithm can gain higher accuracy compared with existing comparison algorithms.

Staphylococcal phosphatidylinositol-specific phospholipase C potentiates lung injury via complement sensitisation.

Staphylococcus aureus is frequently isolated from patients with community-acquired pneumonia and acute respiratory distress syndrome (ARDS). ARDS is associated with staphylococcal phosphatidylinositol-specific phospholipase C (PI-PLC); however, the role of PI-PLC in the pathogenesis and progression of ARDS remains unknown. Here, we showed that recombinant staphylococcal PI-PLC possesses enzyme activity that causes shedding of glycosylphosphatidylinositol-anchored CD55 and CD59 from human umbilical vein endothelial cell surfaces and triggers cell lysis via complement activity. Intranasal infection with PI-PLC-positive S. aureus resulted in greater neutrophil infiltration and increased pulmonary oedema compared with a plc-isogenic mutant. Although indistinguishable proinflammatory genes were induced, the wild-type strain activated higher levels of C5a in lung tissue accompanied by elevated albumin instillation and increased lactate dehydrogenase release in bronchoalveolar lavage fluid compared with the plc- mutant. Following treatment with cobra venom factor to deplete complement, the wild-type strain with PI-PLC showed a reduced ability to trigger pulmonary permeability and tissue damage. PI-PLC-positive S. aureus induced the formation of membrane attack complex, mainly on type II pneumocytes, and reduced the level of CD55/CD59, indicating the importance of complement regulation in pulmonary injury. In conclusion, S. aureus PI-PLC sensitised tissue to complement activation leading to more severe tissue damage, increased pulmonary oedema, and ARDS progression.

Susceptibility rates of clinically important bacteria collected from intensive care units against colistin, carbapenems, and other comparative agents: results from Surveillance of Multicenter Antimicrobial Resistance in Taiwan (SMART).

OBJECTIVES: This study aimed to determine the in vitro susceptibility of commonly encountered Gram-negative bacilli (GNB) recovered from patients admitted to intensive care units (ICUs) in Taiwan against colistin, carbapenems, and other comparative agents. METHODS: In total, 758 nonduplicate GNB isolates were obtained from clinical specimens of ICU patients at seven medical centers in 2016. Minimum inhibitory concentrations (MICs) were determined using the Vitek 2 susceptibility system. The reference broth-microdilution method was performed to determine MICs of colistin. Five main carbapenemase genes among carbapenem-non-susceptible GNB and mcr-1-mcr5 genes among colistin non-wild-type or -resistant isolates were determined. RESULTS: After exclusion 38 Proteus mirabilis and 13 Morganella morganii spp. among 361 Enterobacteriaceae isolates, 34 (9.4%) isolates were carbapenem-insusceptible, 91.1% (n=31) were colistin wild type, and three and one Klebsiella pneumoniae isolates carried bla KPC and bla OXA48-like, respectively. Carbapenem-insusceptible isolates were found in 23.4% (30 of 128) and 63.0% (87 of 138) of isolates of the Pseudomonas aeruginosa and Acinetobacter baumannii complex, respectively. mcr-1 was detected in two (1.8%) Enterobacter cloacae isolates. Very major errors between two methods of susceptibility to colistin were found in 1.5% of K. pneumoniae, 27.5% of E. cloacae, 4.7% of P. aeruginosa, and 10.1% of A. baumannii complex isolates. CONCLUSION: In this study, 8.7% of Enterobacteriaceae isolates from ICUs were not susceptible to carbapenem, and bla KPC and bla OXA48-like were found among three and one carbapenem-insusceptible K. pneumoniae isolates, respectively. Colistin MICs determined by Vitek 2 were not reliable, especially for E. cloacae and A. baumannii complex isolates.

Evaluation of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry Bruker Biotyper for identification of Penicillium marneffei, Paecilomyces species, Fusarium solani, Rhizopus species, and Pseudallescheria boydii.

We evaluated the performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), the MALDI Bruker Biotyper system (microflex LT; Bruker Daltonik GmbH, Bremen, Germany), on the identification of 50 isolates of clinically encountered molds, including Penicillium marneffei (n = 28), Paecilomyces species (n = 12), Fusarium solani (n = 6), Rhizopus species (n = 3), and Pseudallescheria boydii (n = 1). The isolates were identified to species levels by sequence analysis of the internal transcribed spacer (ITS) regions using primers ITS1 and ITS4. None of the 28 genetically well characterized isolates of P. marneffei were identified as P. marneffei by MALDI-TOF MS, because P. marneffei was not present in either Bruker general library (DB 5627) or Bruker filamentous fungi library V1.0. However, the rate of accurate identification as P. marneffei (score value ≥ 2.000) was 85.7% based on newly created database from one P. marneffei strain (NTUH-3370) by MALDI Biotyper system. Sequencing analysis of these 22 non-P. marneffei isolates of molds revealed seven Paecilomyces variotii, six F. solani, four Paecilomyces lilacinus, and one each of Paecilomyces sinensis, Rhizopus arrhizus, R. oryzae, R. microspores, and P. boydii. Although all the seven P. variotii isolates, four of the six F. solani, two of the four P. lilacinus, and two of the three isolates of Rhizopus species, and the P. boydii isolate had concordant identification results between MALDI-TOF MS and sequencing analysis, the score values of these isolates were all of <1.700. This study indicated that the MALDI Bruker Biotyper is ineffective for identifying P. marneffei and other unusual molds because of the current database limitations. Therefore, it is necessary to continuously update the MALDI-TOF MS databases.

Clinical features and antimicrobial susceptibility trends in Citrobacter freundii bacteremia.

This retrospective study investigated the clinical characteristics and antimicrobial susceptibilities of 36 cases of Citrobacter freundii bacteremia treated at the Taipei Veterans General Hospital from 1996 through 1999. The results showed that the predominant infection site was the intraabdominal region and the mortality was 22%. The resistance of C. freundii to most third-generation cephalosporins and broad-spectrum penicillins increased in both nosocomial and community-acquired C. freundii bacteremia during the study period, and the strategy of using a combination of antimicrobial agents to treat C. freundii infection was effective. This study also demonstrated the importance of appropriate antimicrobial therapy to the successful outcome of C. freundii bacteremia. The guidelines of the National Nosocomial Infections Surveillance System were followed to determine trends of resistance of C. freundii to various antimicrobial agents. The resistance of C. freundii to antibiotics in 1999 (n = 10), compared with the period from 1996 through 1998 (n = 26), increased 66% for ciprofloxacin, 36% for ticarcillin/clavulanate, 70% for piperacillin/tazobactam, and 62.8% for piperacillin, but remained uniformly susceptible to imipenem/cilastatin and the new fluoroquinolone (levofloxacin). This increase in resistance was attributable to the use of third-generation cephalosporin instead of first-generation cephalosporins. These findings indicate the need for new measures to facilitate the appropriate choice of antimicrobial agents for patients with C. freundii bacteremia.