Revealing the mechanism: the influence of Baicalin on M1/M2 and Th1/Th2 imbalances in mycoplasma gallisepticum infection.

Mycoplasma gallisepticum (MG) is a pathogen that induces chronic respiratory illnesses in chickens, leading to tracheal and lung injury, and eliciting immune reactions that support sustained colonization. Baicalin, a compound found in scutellaria baicalensis, exhibits anti-inflammatory, antioxidant, and antibacterial properties. This study aimed to investigate the potential of baicalin in alleviating lung and cell damage caused by MG by restoring imbalances in M1/M2 and Th1/Th2 differentiation and to explore its underlying mechanism. In this research, a model for M1/M2 polarization induced by MG was initially developed. Specifically, infection with MG at a multiplicity of infection (MOI) of 400 for 6 h represented the M1 model, while infection for 10 h represented the M2 model. The polarization markers were subsequently validated using qRT-PCR, ELISA, and Western blot analysis. Baicalin disrupts the activation of M1 cells induced by MG and has the potential to restore the balance between M1 and M2 cells, thereby mitigating the inflammatory damage resulting from MG. Subsequent studies on MG-infected chickens detected imbalances in M1/M2 and Th1/Th2 differentiation in alveolar lavage fluid, as well as imbalances in macrophages and Th cells in the lung. The M1/Th1 model was exposed to MG for 5 d, while the M2/Th2 model was infected with MG for 7 d. The utilization of both light and electron transmission microscopes revealed that the administration of baicalin resulted in a reduction in the number of M1 cells, a decrease in cytoplasmic vacuoles, restoration of mitochondrial swelling and chromatin agglutination, as well as alleviation of alveolar rupture and inflammatory cell infiltration. Furthermore, baicalin restored MG-induced M1/M2 and Th1/Th2 imbalances and inhibited the phosphorylation of p38 and p65 proteins, thereby hindering the activation of the TLR4-p38 MAPK/NF-κB pathway. This study provides insights into the potential long-term effects of baicalin in MG infection and offers a theoretical basis for practical applications.

Lactobacillus salivarius ameliorates Mycoplasma gallisepticum-induced inflammation via the JAK/STAT signaling pathway involving respiratory microbiota and metabolites.

Mycoplasma gallisepticum (MG) can cause chronic respiratory disease (CRD) in chickens, which has a significant negative economic impact on the global poultry sector. Respiratory flora is the guardian of respiratory health, and its disorder is closely related to respiratory immunity and respiratory diseases. As a common probiotic in the chicken respiratory tract, Lactobacillus salivarius (L. salivarius) has potential antioxidant, growth performance enhancing, and anti-immunosuppressive properties. However, the specific mechanism through which L. salivarius protects against MG infection has not yet been thoroughly examined. This study intends to investigate whether L. salivarius could reduce MG-induced tracheal inflammation by modulating the respiratory microbiota and metabolites. The results indicated that L. salivarius reduced MG colonization significantly and alleviated the anomalous morphological changes by using the MG-infection model. L. salivarius also reduced the level of Th1 cell cytokines, increased the level of Th2 cell cytokines, and ameliorated immune imbalance during MG infection. In addition, L. salivarius improved the mucosal barrier, heightened immune function, and suppressed the Janus kinase/Signal transducer, and activator of transcription (JAK/STAT) signaling pathway. Notably, MG infection changed the composition of the respiratory microbiota and metabolites, and L. salivarius therapy partially reversed the aberrant respiratory microbiota and metabolite composition. Our results highlighted that these findings demonstrated that L. salivarius played a role in MG-mediated inflammatory damage and demonstrated that L. salivarius, by altering the respiratory microbiota and metabolites, could successfully prevent MG-induced inflammatory injury in chicken trachea.

The antagonism mechanism of astilbin against cadmium-induced injury in chicken lungs via Treg/Th1 balance signaling pathway.

The purpose of this study was to investigate the effect of Treg/Th1 imbalance in cadmium-induced lung injury and the potential protective effect of astilbin against cadmium-induced lung injury in chicken. Cadmium exposure significantly decreased T-AOC and GSH-Px levels and SOD activity in the chicken lung tissues. In contrast, it significantly increased the MDA and NO levels. These results indicate that cadmium triggers oxidative stress in lungs. Histopathological analysis revealed that cadmium exposure further induced infiltration of lymphocytes in the chicken lungs, indicating that cadmium causes pulmonary damage. Further analysis revealed that cadmium decreased the expression of IL-4 and IL-10 but increased those of IL-17, Foxp3, TNF-α, and TGF-ß, indicating that the exposure of cadmium induced the imbalance of Treg/Th1. Moreover, cadmium adversely affected chicken lung function by activating the NF-kB pathway and inducing expression of genes downstream to these pathways (COX-2, iNOS), associated with inflammatory injury in the lung tissue. Astilbin reduced cadmium-induced oxidative stress and inflammation in the lungs by increasing antioxidant enzyme activities and restoring Treg/Th1 balance. In conclusion, our results suggest that astilbin treatment alleviated the effects of cadmium-mediated lung injury in chickens by restoring the Treg/Th1 balance.

Research on directional rock blasting based on different slotted pipe materials of the combined charge structure.

For shaped charge blasting projects in mining, civil engineering, and similar fields, it is proposed to modify the charge structure by combining slotted tubes and shaped charge liners to obtain a new type of charge structure. This aims to achieve directional rock breaking through the focused action of the shaped charge. The influence of different slotted pipe materials on the directional rock-breaking effect of concentrated energy using a new charge structure is explored through theoretical analysis combined with model test study, high-speed camera, stress-strain gauge, and other equipment. A comparison is made between slotted pipes made of aluminum, kraft paper, and PVC, with the cutting width of 2 mm. Based on the characteristics of the cracks formed after blasting, the new charge structure made of aluminum slotted pipe produces a penetrating crack that is almost consistent with the pre-cracking direction. Based on the corresponding characteristics of successively released blasting energy, the guiding and convergence effect of the new charge structure made of aluminum slotted pipe on the explosion energy is greater than that of the new charge structure made of the other two types of slotted pipe material. According to the strain data measured after blasting, the peak arrival time of the strain peak in the direction of the slotted pipe on one side of the shaped hood is shorter than that in the other two directions, and the peak strain is greater than that in the other two directions while having a better energy gathering effect. Based on the findings, the new charge structure with directional energy concentration has a damage reduction effect. Furthermore, the material of aluminum slotted pipe is found to be better than PVC slotted pipe, whereas the material of PVC slotted pipe is better than kraft paper slotted pipe in achieving directional rock breaking.

Methylsulfonylmethane ameliorates inflammation via NF-κB and ERK/JNK-MAPK signaling pathway in chicken trachea and HD11 cells during Mycoplasma gallisepticum infection.

Mycoplasma gallisepticum (MG) is an avian pathogen that commonly causes respiratory diseases in poultry. Methylsulfonylmethane (MSM) is a sulfur-containing natural compound that could alleviate inflammatory injury through its excellent anti-inflammatory and antioxidant properties. However, it is still unclear whether MSM prevents MG infection. The purpose of this study is to determine whether MSM has mitigative effects on MG-induced inflammatory injury in chicken and chicken like macrophages (HD11 cells). In this research, White Leghorn chickens and HD11 cells were used to build the MG-infection model. Besides, the protective effects of MSM against MG infection were evaluated by detecting MG colonization, histopathological changes, oxidative stress and inflammatory injury of trachea, and HD11 cells. The results revealed that MG infection induced inflammatory injury and oxidative stress in trachea and HD11 cells. However, MSM treatment significantly ameliorated oxidative stress, partially alleviated the abnormal morphological changes and reduced MG colonization under MG infection. Moreover, MSM reduced the mRNA expression of proinflammatory cytokines-related genes and decreased the number of death cells under MG infection. Importantly, the protective effects of MSM were associated with suppression of nuclear factor-kappa B (NF-κB) and extracellular signal-related kinases (ERK)/Jun amino terminal kinases (JNK)-mitogen-activated protein kinases (MAPK) pathway in trachea and HD11 cells. These results proved that MSM has protective effects on MG-induced inflammation in chicken, and supplied a better strategy for the protective intervention of this disease.

Dietary administration of Bacillus subtilis KC1 improves growth performance, immune response, heat stress tolerance, and disease resistance of broiler chickens.

The purpose of the present study was to evaluate the probiotic properties of Bacillus subtilis KC1 as a feed additive in the poultry feed. Effects of the Bacillus subtilis supplementation on growth performance, heat-stress tolerance, resistance to Mycoplasma gallisepticum (MG) and Salmonella Pullorum challenge of broilers were determined. The protective effects of the Bacillus subtilis on liver function and immune response of broilers challenged with Aflatoxin B1 (AFB1) were also scrutinized. The results showed that the Bacillus subtilis supplementation could improve growth performance, increased body weight, relative weight of the immune organ and dressing percentage, and decrease feed conversion ratio. In addition, the Bacillus subtilis supplementation alleviated adverse effects caused by heat stress, MG, and Salmonella Pullorum challenge. Furthermore, the Bacillus subtilis supplementation resulted in improved liver function and enhanced immune response of broilers challenged with AFB1. In conclusion, these results suggested a tremendous potential of Bacillus subtilis KC1 as a feed additive in the poultry feed.

Mycoplasma gallisepticum induced inflammation-mediated Th1/Th2 immune imbalance via JAK/STAT signaling pathway in chicken trachea: Involvement of respiratory microbiota.

The respiratory microbiota plays a significant role in the host defense against Mycoplasma gallisepticum (MG) infection. The results showed that MG infection changed respiratory microbiota composition, which lead to the tracheal inflammation injury and oxidative stress. MG infection significantly induced immunosuppression in chickens at day 3 and 5 post-infection. In addition, MG infection increased the expressions of pro-inflammatory cytokines in tracheal tissues and activated TLR4 mediated JAK/STAT signaling pathway at day 3 post-infection compared to the control group. Meanwhile, the expressions of pro-inflammatory cytokines were decreased and the expressions of JAK/STAT signaling pathway were decreased at day 5 and day 7 post-infection. On the contrary, the expressions of anti-inflammatory cytokines were significantly decreased at day 3 post-infection and were increased at day 5 and day 7 post-infection in the MG infection group. The antibiotic cocktail group received the respiratory microbiota from the MG infection group, which induced inflammatory injury and oxidative stress, induced mucosal barrier damage by down regulating tight junction-related genes and altered the expressions of mucin, which could be the possible causes of dysregulated immune responses. Importantly, the expressions of pro-inflammatory cytokines were significantly decreased and TLR4 mediated JAK/STAT signaling pathway was downregulated at day 1 and 3 post-transplantation. While, respiratory microbiota transplanted from MG infection significantly increased the expressions of pro-inflammatory cytokines and activated JAK/STAT signaling at day 7 post-transplantation. These results highlighted the role of respiratory microbiota in MG-induced tracheal inflammation injury, and offered a new strategy for the preventive intervention of this disease.

Pharmacokinetic/pharmacodynamic profiles of baicalin against Mycoplasma gallisepticum in an in vivo infection model.

Mycoplasma gallisepticum (M. gallisepticum), a devastating avian pathogen that commonly causes chronic respiratory disease in chicken, is responsible for tremendous economic losses to the poultry industry. Baicalin is the main constituent of Scutellaria baicalensis that shows potential therapeutic effects against M. gallisepticum. However, the pharmacokinetic/pharmacodynamics (PK/PD) profiles of baicalin against M. gallisepticum are not well understood. The main objective of the present study was to determine the relationship between the PK/PD index and efficacy of baicalin in the M. gallisepticum infection model in chickens. The experiments were carried out on 10-day-old chickens that were challenged with M. gallisepticum in the bilateral air sacs. While, baicalin was orally administrated once in a day for 3 consecutive days, started from d 3 postinfection. Ultra-performance liquid chromatography (UPLC) was used to evaluate the PK parameters of baicalin at doses of 200, 400, and 600 mg/kg in M. gallisepticum-infected chickens. Real-time PCR (RT-PCR) was used for the quantitative detection of M. gallisepticum in lungs. The PK and PD data were fitted to WinNonlin software to evaluate the PK/PD profiles of baicalin against M. gallisepticum. The minimum inhibitory concentration (MIC) of baicalin against M. gallisepticum strain Rlow was 31.25 µg/mL. The in vivo data suggested that baicalin concentration in the lung tissues was higher than plasma (1.21-1.73 times higher). The ratios of AUC24h/MIC of baicalin against bacteriostatic, bactericidal, and eradication were 0.62, 1.33, and 1.49 h, respectively. In conclusion, these results provided potential reference for future clinical dose selection of baicalin and evaluation of susceptibility breakpoints.

METTL3-mediated M6A methylation modification is involved in colistin-induced nephrotoxicity through apoptosis mediated by Keap1/Nrf2 signaling pathway.

Colistin is a cationic polypeptide antibiotic. Despite its nephrotoxicity, it is still widely used as a last-line antibiotic against infection worldwide with the emergence of multi-drug resistant Gram-negative bacilli. N-methyladenosine (m6A) methylation-mediated degradation of RNA is essential for kidney development. However, m6A methylation impacts not only RNA stability, but also other RNA metabolism processes. How RNA decay affects the nephrotoxicity of colistin is largely unknown. Therefore, in this study, we verified that colistin could induce mouse kidney apoptosis through some apoptotic indicators, and confirmed the relationship between methylation and apoptosis through the detection of m6A methylation, thus elucidating the potential mechanism of colistin nephrotoxicity. The results showed that the renal tubule dilation and tubular structure were observed in the colistin group, and the oxidative stress index and ATPase activities were significantly different from those in the control group. Under electron microscope, the kidney in colistin group showed typical apoptotic morphological changes such as nuclear pyknosis, chromatin edge aggregation, and intact nuclear membrane, accompanied by significant changes in apoptosis-related genes. The level of m6A in the colistin group was significantly decreased, accompanied by downregulation of METTL3 mRNA and protein levels, and METTL3 was significantly correlated with apoptotic gene proteins. Data from this study suggested that m6A methylation was involved in oxidative stress-mediated apoptosis in the mechanism of colistin nephrotoxicity.

Baicalin attenuated Mycoplasma gallisepticum-induced immune impairment in chicken bursa of fabricius through modulation of autophagy and inhibited inflammation and apoptosis.

BACKGROUND: Mycoplasma gallisepticum (MG) is the primary etiologic agent of chronic respiratory disease in poultry. However, the mechanism underlying MG-induced immune dysregulation in chicken is still elusive. Baicalin shows excellent anti-bacterial, anti-inflammatory, anti-carcinogenic and anti-viral properties. In the present study, the preventive effects of baicalin against immune impairment in chicken bursa of fabricius (BF) were studied in an MG infection model. RESULTS: Histopathological examination showed increased inflammatory cell infiltrations and fragmented nuclei in the model group. Ultrastructural analysis revealed the phenomenon of apoptosis in bursal cells, along with the deformation of mitochondrial membrane and swollen mitochondria in the model group. However, these abnormal morphological changes were partially alleviated by baicalin. Meanwhile, baicalin treatment attenuated the level of proinflammatory cytokines, and suppressed nuclear factor-kappa B expression at both protein and mRNA level. Terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling assay showed extensive apoptosis in BF in the model group. The mRNA and protein expression levels of apoptosis-related genes were upregulated in BF, while baicalin treatment significantly alleviated apoptosis in BF. In addition, alterations in mRNA and protein expression levels of autophagy-related genes and mitochondrial dynamics proteins were significantly alleviated by baicalin. Moreover, baicalin treatment significantly attenuated MG-induced decrease in CD8+ cells and reduced bacterial load in chicken BF compared to the model group. CONCLUSIONS: These results suggested that baicalin could effectively inhibit MG-induced immune impairment and alleviate inflammatory responses and apoptosis in chicken BF. © 2020 Society of Chemical Industry.

Baicalin inhibits inflammation caused by coinfection of Mycoplasma gallisepticum and Escherichia coli involving IL-17 signaling pathway.

Coinfection of Mycoplasma gallisepticum (MG) and Escherichia coli (E. coli) is frequently reported in poultry farms. Baicalin possess various pharmacological properties such as anti-inflammatory, anticancer, and antioxidant, etc. However, the protective effects of baicalin against coinfection of MG and E. coli are still elusive. In this study, baicalin (450 mg/kg) treatment was started on day 13 after infection and continued for 5 d. Histopathological examination, qRT-PCR, ELISA, and molecular docking technique were used to evaluate the effects of baicalin on MG and E. coli coinfection in chicken lung and trachea. The results showed that coinfection caused severe lesions in the lung and tracheal tissues. However, baicalin treatment partially alleviated these lesions in coinfection group. Histopathological examination showed the alveolar spaces and mucosal layer thickening was restored and cilia gradually recovered with baicalin treatment compared in coinfection group and MG-infection group. Meanwhile, IL-17 singling pathway-related genes were significantly reduced (P < 0.05) in baicalin treatment group in lung, including IL-17C, TRAF6, NF-κB, CXCL1, CXCL2, MMP1, GM-CSF, and MUC5AC. The activities of cytokines and chemokines (CXCL1, CXCL2, MMP1, GMCSF, and MUC5AC) were decreased significantly (P < 0.05) in baicalin-treated group. The molecular docking of baicalin and NF-κB showed the highest fitness score and interaction. From these results, it has been suggested that baicalin proved effective against coinfection of MG and E. coli in chicken and provided scientific basis for further dose-response and drug-target interaction studies.

Population pharmacokinetics for danofloxacin in the intestinal contents of healthy and infected chickens.

Avian pathogenic Escherichia coli could cause localized and systemic infection in the poultry, and danofloxacin is usually used to treat avian colibacillosis through oral administration. To promote prudent use of danofloxacin and reduce the emergence of drug-resistant E. coli strains, it is necessary to understand the population pharmacokinetics (PopPK) of danofloxacin in chicken intestines. In this study, reversed-phase high performance liquid chromatography (HPLC) with fluorescence detection was used to detect the concentrations of danofloxacin in the contents of duodenum, jejunum, and ileum of the healthy and infected chickens after single oral administration (5 mg/kg body weight). Then, the PopPK of danofloxacin in intestines were analyzed using NONMEM software. As a result, a two-compartment PK model best described the time-concentration profile of duodenal, jejunal, and ileal contents. Interestingly, absorption rate (Ka ), distribution volume (V), and clearance (CL) for danofloxacin from duodenal, jejunal to ileal contents were sequentially decreased in the healthy chickens. However, the trend of Ka , V, and CL of danofloxacin was changed dramatically in the intestine of infected chickens. Ka and V of danofloxacin in the jejunum were higher than in the ileum and duodenum. Compared with healthy chickens, Ka and V of danofloxacin in the duodenum decreased significantly, while increased in jejunum, respectively. It has been noted that Ka decreased and V increased in the ileum of infected chickens. Besides, CL in the duodenum, jejunum, and ileum of infected chickens was, respectively, lower than those of healthy chickens. Interestingly, the relative bioavailability (F) of danofloxacin in the ileum was relatively higher in both healthy and infected chickens. In addition, F in the duodenal, jejunal, and ileal contents of infected chickens was respectively higher than healthy chickens. In summary, the PopPK for danofloxacin in infected chicken intestines was quite different from healthy chickens. The absorption, distribution, and clearance of danofloxacin in healthy chickens decreased from duodenum to jejunum and to ileum. Moreover, the pharmacokinetic characteristics in the intestine of infected chickens changed significantly, and the pharmacokinetic characteristics in the ileum can be used as a representative of all intestinal segments.

Preventive effects of nerve growth factor against colistin-induced autophagy and apoptosis in PC12 cells.

In this study, the preventive effects of NGF against colistin-induced autophagy and apoptosis in PC12 cells have been investigated. Fluorescence microscopy, real-time PCR, transmission electron microscopy (TEM), flow cytometery, and western blotting technique were used. The results showed that large amounts of autophagosomes and apoptotic markers were triggered by colistin. Consistently, a significant increase has been noted at mRNA and protein levels in autophagy and apoptosis-related genes. Besides, TEM analysis showed that autophagic vacuoles were obvious at 12 h, while nuclear chromatin condensation and edge accumulation were clearly seen at 24 h in colistin alone group. Importantly, the visual autophagy and apoptosis were markedly reduced with NGF treatment in a dose-dependent manner. Moreover, colistin-induced reduction in mitochondrial membrane potential was partly attenuated by NGF in a dose dependent manner. In summary, NGF ameliorated colistin-induced apoptosis and autophagy, and partially recovered MMP in PC12 cells.

Baicalin Attenuates Mycoplasma gallisepticum-Induced Inflammation via Inhibition of the TLR2-NF-κB Pathway in Chicken and DF-1 Cells.

BACKGROUND: Previous reports demonstrated that baicalin possesses potential anti-inflammatory properties. The present study was conducted to determine the effects of baicalin against inflammatory responses in chicken and DF-1 cells infected with Mycoplasma gallisepticum (MG). METHODS: An MG infection model was developed in chickens to study the anti-inflammatory mechanism of baicalin. Baicalin was mixed in water at a dose of 450 mg/kg per day, and the treatment is continued for 7 consecutive days. Samples were taken at 1, 4, and 7 days post treatment. RESULTS: By using transmission electron microscopy, ultrastructure of lung and tracheal cells has been examined. It can be seen that the cilia cells in the MG-infected group have pyknosis, degeneration, and necrosis. In the lung tissues, alveolar type-I epithelial cells were severely damaged. In the baicalin-treated group, cilia were swollen, mushroom-shaped edema bubbles formed on the apex, and fused together. Alveolar type I epithelial cells injury was significantly reduced. Compared to MG-infection group, the levels of proinflammatory cytokines IL-1ß and TNF-α were significantly decreased (P < 0.01). The corresponding proteins TLR2 and P-p65 decreased in the baicalin-treated group after 1 (p > 0.05), 4 (p < 0.05), and 7 days (p < 0.05), respectively. CONCLUSION: The results showed that baicalin can interfere with inflammatory injury by suppressing the release of inflammatory cytokines IL-1ß and TNF-α during MG infection both in vivo and in vitro. Meanwhile, baicalin suppressed TLR2-NFκB signaling pathway by inhibiting the phosphorylation of p65 and IκB, thereby affecting the expression of inflammatory factors. The results suggested that baicalin acts as a potential anti-inflammatory agent against MG infection in chicken and DF-1 cells.

A Dual Role of P53 in Regulating Colistin-Induced Autophagy in PC-12 Cells.

This study aimed to investigate the mechanism of p53 in regulating colistin-induced autophagy in PC-12 cells. Importantly, cells were treated with 125 µg/ml colistin for 12 and 24 h after transfection with p53 siRNA or recombinant plasmid. The hallmarks of autophagy and apoptosis were examined by real-time PCR and western blot, fluorescence/immunofluorescence microscopy, and electron microscopy. The results showed that silencing of p53 leads to down-regulation of Atg5 and beclin1 for 12 h while up-regulation at 24 h and up-regulation of p62 noted. The ratio of LC3-II/I and autophagic vacuoles were significantly increased at 24 h, but autophagy flux was blocked. The cleavage of caspase3 and PARP (poly ADP-ribose polymerase) were enhanced, while PC-12-sip53 cells exposed to 3-MA showed down-regulation of apoptosis. By contrast, the expression of autophagy-related genes and protein reduced in p53 overexpressing cells following a time dependent manner. Meanwhile, there was an increase in the expression of activated caspase3 and PARP, condensed and fragmented nuclei were evident. Conclusively, the data supported that silencing of p53 promotes impaired autophagy, which acts as a pro-apoptotic induction factor in PC-12 cells treated with colistin for 24 h, and overexpression of p53 inhibits autophagy and accelerates apoptosis. Hence, it has been suggested that p53 could not act as a neuro-protective target in colistin-induced neurotoxicity.

Colistin-induced autophagy and apoptosis involves the JNK-Bcl2-Bax signaling pathway and JNK-p53-ROS positive feedback loop in PC-12 cells.

Our recent study demonstrated neurotoxicity of colistin-induced autophagy and apoptosis in PC-12 cells, and that autophagy reached peak level at 12 h. In this study, we scrutinized the role of JNK in colistin-induced neurotoxicity and demonstrated the relationship among JNK, p53 and ROS in colistin treated PC-12 cells. Colistin-induced autophagy and apoptosis by JNK inhibition/activation were examined by western blotting, electron microscopy, and immunofluorescence/fluorescence microscopy. The results indicated that colistin induced JNK activation reached peak level at 12 h, while the highest levels of p-Bcl2/Bcl2 were observed at 12 h and Bax/Bcl2 significantly increased in a time-dependent manner. In PC-12 cells, inhibition of JNK by SP600125 (JNK inhibitor) resulted in significantly lower levels of autophagy upon colistin treatment, depending on the expression levels of Beclin1, LC3-II, p62 degradation and reduction in the number of autophagic vacuoles. In contrast, anisomycin pretreatment PC-12 cells led to upregulated autophagy. Especially, the highest levels of Beclin1 and p-Bcl2/Bcl2 were observed at 6 h, and Bax/Bcl2, cleaved-caspase3 and cleaved-PARP significantly increased in a time-dependent manner. The results revealed that JNK activation mediated autophagy and apoptosis related to Beclin1-Bcl2 and Bax-Bcl2 complex in colistin-treated PC-12 cells. Silencing of p53 by siRNA before colistin treatment substantially reduced ROS production and transactivated JNK in PC-12 cells. Moreover, activation of JNK increased ROS generation in PC-12 cells. In conclusion, colistin-induced autophagy and apoptosis is correlated to JNK-Bcl2-Bax signaling pathway, and an interaction effect found between intracellular ROS level and JNK-p53 signaling pathway in apoptosis.

TLR2 mediates autophagy through ERK signaling pathway in Mycoplasma gallisepticum-infected RAW264.7 cells.

Toll-like receptor 2 (TLR2) plays a crucial role in early innate immune response of host to various microorganisms. Mycoplasma gallisepticum (MG) is one of the major pathogen that can cause chronic respiratory diseases in chickens, but the molecular mechanism of MG infection still remained unclear. In this study, we examined the typical hallmarks of autophagy and multiple signaling pathways by western blot, immunofluorescence microscopy and electron microscopy. The results indicated that infection of mouse macrophage cell line RAW264.7 with MG activated autophagy and mitogen-activated protein kinases (MAPKs). Silencing of TLR2 by siRNA substantially down-regulated MG-triggered autophagy in macrophages, and markedly reduced MG-induced extracellular regulated protein kinase (ERK) in macrophages but did not down-regulate c-Jun N-terminal kinase (JNK) and p38. Importantly, in macrophages, inhibition of ERK by PD98059 (ERK inhibitor) also significantly attenuated the level of autophagy upon MG infection, and the simultaneous treatment of TLR2 siRNA and PD98059 showed a similar effect on MG-induced autophagy as compared with TLR2 siRNA treatment alone. These findings thus demonstrate that TLR2 may mediate MG-induced autophagy through ERK signaling pathway in macrophage.

A velocity probe-based method for continuous detonation and shock measurement in near-field underwater explosion.

A new velocity probe which permits recording the time history of detonation and shock waves has been developed by improving the commercial on principle and structure. A method based on the probe is then designed to measure the detonation velocity and near-field shock parameters in a single underwater explosion, by which the oblique shock wave front of cylindrical charges and the peak pressure attenuation curve of spherical explosive are obtained. A further derivation of detonation pressure, adiabatic exponent, and other shock parameters is conducted. The present method offers a novel and reliable parameter determination for near-field underwater explosion.

Attenuation of Porphyromonas gingivalis oral infection by α-amylase and pentamidine.

The Porphyromonas gingivalis bacterium is one of the most influential pathogens in oral infections. In the current study, the antimicrobial activity of α-amylase and pentamidine against Porphyromonas gingivalis was evaluated. Their in vitro inhibitory activity was investigated with the agar overlay technique, and the minimal inhibitory and bactericidal concentrations were determined. Using the bactericidal concentration, the antimicrobial actions of the inhibitors were investigated. In the present study, multiple techniques were utilized, including scanning electron microscopy (SEM), general structural analysis and differential gene expression analysis. The results obtained from SEM and bactericidal analysis indicated a notable observation; the pentamidine and α-amylase treatment destroyed the structure of the bacterial cell membranes, which led to cell death. These results were used to further explore these inhibitors and the mechanisms by which they act. Downregulated expression levels were observed for a number of genes coding for hemagglutinins and gingipains, and various genes involved in hemin uptake, chromosome replication and energy production. However, the expression levels of genes associated with iron storage and oxidative stress were upregulated by α-amylase and pentamidine. A greater effect was noted in response to pentamidine treatment. The results of the present study demonstrate promising therapeutic potential for α-amylases and pentamidine. These molecules have the potential to be used to develop novel drugs and broaden the availability of pharmacological tools for the attenuation of oral infections caused by Porphyromonas gingivalis.