Mycoplasma hyopneumoniae inhibits the unfolded protein response to prevent host macrophage apoptosis and M2 polarization.

Enzootic pneumonia caused by Mycoplasma hyopneumoniae (M. hyopneumoniae) has inflicted substantial economic losses on the global pig industry. The progression of M. hyopneumoniae induced-pneumonia is associated with lung immune cell infiltration and extensive proinflammatory cytokine secretion. Our previous study established that M. hyopneumoniae disrupts the host unfolded protein response (UPR), a process vital for the survival and immune function of macrophages. In this study, we demonstrated that M. hyopneumoniae targets the UPR- and caspase-12-mediated endoplasmic reticulum (ER)-associated classical intrinsic apoptotic pathway to interfere with host cell apoptosis signaling, thereby preserving the survival of host tracheal epithelial cells (PTECs) and alveolar macrophages (PAMs) during the early stages of infection. Even in the presence of apoptosis inducers, host cells infected with M. hyopneumoniae exhibited an anti-apoptotic potential. Further analyses revealed that M. hyopneumoniae suppresses the three UPR branches and their induced apoptosis. Interestingly, while UPR activation typically drives host macrophages toward an M2 polarization phenotype, M. hyopneumoniae specifically obstructs this process to maintain a proinflammatory phenotype in the host macrophages. Overall, our findings propose that M. hyopneumoniae inhibits the host UPR to sustain macrophage survival and a proinflammatory phenotype, which may be implicated in its pathogenesis in inducing host pneumonia.

Non-Communication Decentralized Multi-Robot Collision Avoidance in Grid Map Workspace with Double Deep Q-Network.

This paper presents a novel decentralized multi-robot collision avoidance method with deep reinforcement learning, which is not only suitable for the large-scale grid map workspace multi-robot system, but also directly processes Lidar signals instead of communicating between the robots. According to the particularity of the workspace, we handcrafted a reward function, which considers both the collision avoidance among the robots and as little as possible change of direction of the robots during driving. Using Double Deep Q-Network (DDQN), the policy was trained in the simulation grid map workspace. By designing experiments, we demonstrated that the learned policy can guide the robot well to effectively travel from the initial position to the goal position in the grid map workspace and to avoid collisions with others while driving.

Analysis of plasma migration components in Patrinia villosa (Thunb.) Juss. effective parts by UPLC-Q-TOF-MS.

Patrinia villosa (Thunb.) Juss. (PVJ) is described as pungent, bitter and slightly cold in Chinese medicine, and is associated with the large intestine, stomach and liver meridians. The preliminary experiments of our research team proved that PVJ total flavonoids have excellent inhibitory effects on liver cancer cells. The present experiment uses the UPLC-Q-TOF-MS technology and serum pharmacochemistry methods to analyze the chemical components in vitro and in vivo of PVJ antiliver tumors. A total of 14 chemical components were identified in the total flavonoids extract of PVJ, and it is mainly composed of flavonoids, flavonones, flavonols and phenolic acids. At the same time, seven prototypical components and seven metabolic components were detected in the drug-containing plasma. Hydrocaffeate and scutellarein are the phase I metabolites of caffeic acid and scutellarin, respectively. Sulfated apigenin, sulfated luteolin, sulfated kaempferol and methylated kaempferol are the II phase metabolites of apigenin, luteolin, kaempferol, respectively. The experiment provides a reference for the research and development of antitumor drug candidates, and provides a basis for revealing the bioactive components of PVJ and the antitumor mechanism.

Multiple Pyogenic Granulomas After Burns: Response to Conservative Treatment in Five Children.

We describe five children with multiple pyogenic granulomas after burns that were healed effectively using conservative treatment consisting of 1% povidone-iodine, silver nanoparticle dressing, and antibiotics. No relapse of the lesions was observed from 6 months to 2 years later.

[Differential expression profile of microRNAs between human bone marrow mesenchymal stem cells and sweat gland-like cells].

OBJECTIVE: To explore the differential microRNAs (miRNAs) expression profiles between human bone marrow mesenchymal stem cells (hBM-MSCs) and sweat gland-like cells. METHODS: The hBM-MSCs were induced to differentiate into osteogenic and adipogenic cells by related osteogenic differentiation basal medium and adipogenic differentiation basal medium respectively. And the results were assayed by oil red O and alizarin red stains respectively. When reaching 70%-80% confluence, primarily cultured sweat gland cells were heat-shocked and co-cultured with hBM-MSCs by a Transwell plate to induce the differentiation of hBM-MSCs. After 10 days, the total RNAs of hBM-MSCs and sweat gland-like cells were extracted, purified, labeled, hybridized and analyzed for predicting miRNAs target genes separately. The microarray results were confirmed by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: There were 68 miRNAs of >double differentiation on expression level between hBM-MSCs and sweat gland-like cells. And 19 miRNAs were up-regulated and 49 miRNAs down-regulated. Moreover, there were 13 miRNAs with >5 folds of differential expression level between hBM-MSCs and sweat gland-like cells, including 5 up-regulated miRNAs (miRNA-132-3p, -4467, -4484, -146a-5p and -6126) and 8 down-regulated miRNAs (miRNA-708-5p, -138-5p, -6812-5p, -138-1-3p, -1281, -3157-3p, -4298 and -4459). There were 18 miRNAs related to regulation of the signaling pathways of sweat gland development, including nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase/extracellular regulated protein kinase (MAPK/Erk), Wnt/beta-Catenin (Wnt/ß-Catenin) and ectodysplasin-A1/ectodysplasin-A1 receptor (EDA/EDAR). The results of RT-PCR on miRNA-146a-5p and miRNA-6812-5p had a high concordance with the results of microarray. CONCLUSIONS: There are obvious differentiation miRNAs expression profiles between hBM-MSCs and sweat gland-like cells. And 13 miRNAs >5 folds of differential expression and 18 miRNAs related to sweat gland development may play essential roles in regulated differentiation of hBM-MSCs into sweat gland-like cells.

[DNA methylation of HDAC4 gene affects the trans-differentiation process of hMSCs into sweat gland like cells].

OBJECTIVE: To investigate the changesof DNA methylation in histone deacetylases 4 gene (HDAC4) and its effectduring the trans-differentiation process of human mesenchymal stem cells (hMSCs) into sweat gland like cells (SGLCs). METHODS: Selected cell lines of human mesenchymal stem cells (hMSCs) were cultured and expended in vitro, the third generation ofhMSCs and heat-shocked sweat gland cells were picked up, and were co-culturedwith adding inducible factor in the transwell chamber. The sweat gland like cells (SGLCs)in experiment group and the hMSCs in control group were collected, the changes of DNA methylation degree of CpG dinucleotide sitesin histone deacetylases 4 gene (HDAC4) promotor were detected by methylation specific PCR (MSP)andMaldi-TOF Mass Array. And then, the hMSCs in experiment group were treated with 5-aza-CdR (5-aza-2-deoxycytidine, 10 µmol/L), while the hMSCsin control group were culturedwith PBS at the same time. ThemRNA expressions of HDAC4 gene and carcino-embryonic antigen (CEA)gene in the two groups were measured by RT-PCR. RESULTS: The methylation of HDAC4gene in hMSCs was in high level before induction, the methylation degreeof CpG dinucleotide sites located in cg2463009 was 0.901, and the methylation degree of HDAC4gene in SGLCs was markedly decreased by 37% after induction, which was 0.531. The methylationlevel of CpG dinucleotide sites located in cg14823429was changed from 0.687to 0.386 after induction. The mRNA expression of HDAC4 gene was upregulated in test group after treated with 5-aza-CdR for 48 hours, the mRNA expression of CEA gene related with transdifferentiation was enhanced too at the same term, there was significantly statistic difference compared with control group (P<0.05). CONCLUSIONS: Methylation of HDAC4 gene participates in the regulation of the trans-differentiation of hMSCs into sweet gland like cells.

[Optimization of primary human eccrine sweet glands' isolation in vitro].

OBJECTIVE: To optimize the methods of isolating human eccrine sweat gland cells in vitro so as to get efficiently primary human sweat glands. METHODS: The fresh and normal skin tissue was cut into pieces of microskin about 1mm3 and the following 3 group digestion buffer was applied to isolated gland cells. The digestion buffer of group A was the equivoluminal mixture of Trypsin-Ethylene Diamine Tetraacetic Acid (EDTA) and collagenase-II (2 mg/ml). The digestion buffer of group B was collagenase-II (2 mg/ml) traditionally and group C was Trypsin-EDTA. These three groups were placed into an incubator simultaneously and the emerging time of dissociated sweat glands was calculated. Sweat glands were sorted out and then placed in culture dish. The adherence and the growth of cells were observed. The proliferation index was detected by flow cytometry. The identification of cultured cells was performed by immunocytochemical staining. RESULTS: After digesting 30 min in group A and C, a very few of dissociated sweat glands were emerging. But after digesting for 2 h, there were lots of dissociated sweat glands emerging in group A rather than in group C. The emergence of dissociated sweat glands in group B would require at least 6 hours. After seeded in culture dishes, the sweat glands in group C couldn't adhere to the wall of dish, but the sweat glands in group A and B adhered very well and even grew like paving stones after 9 days. In addition, the proliferation index were (18 ± 4) % and (17 ± 6) % respectively, there was no statistical difference. The results of immunocytochemical staining showed that the cells expressed carcino-embryonic antigen (CEA) and cytokeratin 7(CK7) in group A and B. CONCLUSION: Trypsin-EDTA combined with collagenase-II can shorten the time of isolating sweat gland cells and have no effect on cell activity and proliferation.