Efficient production of single cell protein from biogas slurry using screened alkali-salt-tolerant Debaryomyces hansenii.

Production of single cell protein (SCP) by recovering ammonia nitrogen from biogas slurry shows great potential against protein scarcity and unsustainable production of plant and animal proteins. Herein, a high-alkali-salt-tolerant yeast strain, Debaryomyces hansenii JL8-0, was isolated and demonstrated for high-efficient SCP production. This strain grew optimally at pH 8.50 and 2500 mg/L NH4+-N, and it could efficiently utilize acetate as the additional carbon source. Under optimal conditions, SCP biomass of 32.21 g/L and productivity of 0.32 g/L·h-1 were obtained in fed-batch fermentation. Remarkably, nearly complete (97.40 %) ammonia nitrogen from biogas slurry was recovered, probably due to its high affinity for NH4+-N. Altogether, this strain showed advantages in terms of cell biomass titer, productivity, and yield. A cultivation strategy was proposed by co-culturing D. hansenii with other compatible yeast strains to achieve high-efficient SCP production from biogas slurry, which could be a promising alternative technology for biogas slurry treatment.

[Metabolomics study of Berberidis Radix in intervening ulcerative colitis based on UPLC-Q-TOF-MS].

The effect of Tujia medicine Berberidis Radix on endogenous metabolites in the serum and feces of mice with ulcerative colitis(UC) induced by dextran sulfate sodium(DSS) was analyzed by metabolomics technology to explore the metabolic pathway and underlying mechanism of Berberidis Radix in the intervention of UC. The UC model was induced in mice by DSS. Body weight, disease activity index(DAI), and colon length were recorded. The levels of tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) in colon tissues were determined by ELISA. The levels of endogenous metabolites in the serum and feces were detected by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to characterize and screen differential metabolites. The potential metabolic pathways were analyzed by MetaboAnalyst 5.0. The results showed that Berberidis Radix could significantly improve the symptoms of UC mice and increase the level of the anti-inflammatory factor IL-10. A total of 56 and 43 differential metabolites were identified in the serum and feces, respectively, belonging to lipids, amino acids, fatty acids, etc. After the intervention by Berberidis Radix, the metabolic disorder gradually recovered. The involved metabolic pathways included biosynthesis of phenylalanine, tyrosine, and tryptophan, linoleic acid metabolism, phenylalanine metabolism, and glycerophospholipid metabolism. Berberidis Radix can alleviate the symptoms of mice with DSS-induced UC, and the mechanism may be closely related to the re-gulation of lipid metabolism, amino acid metabolism, and energy metabolism.

HIV Tat-Conjugated Histone H3 Peptides Induce Tumor Cell Death Via Cellular Stress Responses.

Histone H3 is a nucleosome scaffold protein that is involved in a variety of intracellular processes. Aberrant modification of H3 is important in carcinogenesis. In contrast, free histones in cells can act as stimuli to trigger cellular immune responses and cell death. In this study, we linked cell-penetrating peptide HIV Tat to a histone H3 fragment to achieve intracellular delivery in tumor cells. We found that Tat-conjugated histone polypeptides localized to nuclei of lung and breast cancer cells and caused cell death. A trans-configured Tat sequence displayed dramatically improved peptide half-life and cytotoxicity. Mechanistic studies demonstrated that treatment with the peptides significantly elevated mitogen-activated protein kinase (MAPK) signaling, reactive oxygen species (ROS) production, as well as levels of stress-inducible transcription factor ATF3 (activating transcription factor 3) and AP-1 (activating protein-1). Cytotoxicity of the peptide was significantly reduced by inhibition of AP-1 activity and ROS production. These results suggest the potential of Tat-conjugated H3 peptides as antitumor agents to induce cell death via increased cellular stress response by activating p38-MAPK signaling and intracellular ROS production.

Epoxymicheliolide prevents dextran sulfate sodium-induced colitis in mice by inhibiting TAK1-NF-κB pathway and activating Keap1-NRF2 signaling in macrophages.

Ulcerative colitis (UC) is an unspecific colorectal inflammation associated with macrophages overactivation. Therefore, macrophage-targeted treatment has been considered a promising strategy for UC therapy. Epoxymicheliolide (EMCL) is a compound from Aucklandia lappa Decne, a TCM for treating gastrointestinal inflammatory diseases. The purpose of this study is to investigate the therapeutic effect of EMCL on DSS-induced mice colitis through the anti-inflammatory activity on macrophages and its underlying mechanisms. We found that EMCL inhibited the release of NO and PGE2 by down-regulating the expression of iNOS and COX2, while suppressed the expression of IL-1ß, IL-6, and TNF-α in LPS-stimulated RAW264.7 macrophages. EMCL also inhibited NO production in LPS-activated peritoneal macrophages and TNFα-stimulated RAW264.7 cells. Moreover, EMCL blocked the phosphorylation of TAK1, IKKα/ß, and IκBα, as well as IκBα degradation, thereby inhibiting the NF-κB pro-inflammatory signaling. Furthermore, EMCL decreased the intracellular ROS, by activating the NRF2 antioxidant pathway. CETSA and molecular docking showed that EMCL might form a covalent bond with Cys174 of TAK1 or Cya151 of Keap1, which may contribute to EMCL-mediated actions. Additionally, a thiol donor ß-mercaptoethanol obviously abolished EMCL-mediated actions in vitro, suggesting the crucial role of the α, γ-unsaturated lactone of EMCL on its anti-inflammatory effects. Furthermore, EMCL not only ameliorated symptoms of colitis and colon barrier injury, but also decreased the levels of pro-inflammatory cytokines, MPO, NO, and MDA in DSS-challenged mice. Thus, our study demonstrated that EMCL ameliorated UC by targeting NF-κB and Nrf2 pathways, indicating it may server as a promising drug candidate for UC therapy.

Streptomyces xanthii sp. nov. and Streptomyces roseirectus sp. nov. isolated from a Chinese medicinal plant.

Two strains of Actinobacteria, designated CRXT-Y-14T and CRXT-G-22T, were isolated from the healthy leaves and seeds, respectively, of a medicinal plant Xanthium sibiricum. Their taxonomic positions were determined using a polyphasic approach. Strain CRXT-Y-14T produced flexuous chains of smooth-surfaced spores. Strain CRXT-G-22T produced straight chains of smooth-surfaced spores. Their morphological features were consistent with the diagnostic characteristics of members of the genus Streptomyces. The results of 16S rRNA gene sequence analyses indicated two strains represented members of the genus Streptomyces. CRXT-Y-14T shared 99.3, 98.9, 98.8 % sequence similarities to Streptomyces atriruber NRRL B-24165T, Streptomyces avermitilis MA-4680T and Streptomyces davaonensis JCM 4913T, respectively. Whilst CRXT-G-22T exhibited highest similarity to Streptomyces acidiscabies ATCC 49003T (98.9 %). The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that the closest phylogenetic neighbours of strains CRXT-Y-14T and CRXT-G-22T were S. atriruber NRRL B-24165T and S. acidiscabies ATCC 49003T, respectively. The phylogenomic analyses further confirmed the relative relationship between strain CRXT-G-22T and S. acidiscabies ATCC 49003T, but indicated that CRXT-Y-14T could represent a novel species of the genus Streptomyce. However, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between CRXT-Y-14T and strain CRXT-G-22T, between CRXT-Y-14T and S. atriruber NRRL B-24165T, and between CRXT-G-22T and S. acidiscabies ATCC 49003T were 85.4 and 23.2 %, 85.8 and 23.9 % and 89.1 and 34.1 %, respectively, far below the 95~96 and 70 % cut-off points recommended for delineating species. Furthermore, these two novel isolates were distinctly differentiated from their relatives in the genus Streptomyces with respect to phenotypic and chemotaxonomic characteristics. On the basis of these data, CRXT-Y-14T and CRXT-G-22T clearly represent two novel species within the genus Streptomyces, for which the names Streptomyces xanthii sp. nov. (type strain CRXT-Y-14T = MCCC 1K04966T= JCM 34527T) and Streptomyces roseirectus sp. nov. (type CRXT-G-22T = MCCC 1K04979T= JCM 34565T) are proposed.

The regulatory effect of acetylation of HMGN2 and H3K27 on pyocyanin-induced autophagy in macrophages by affecting Ulk1 transcription.

Pyocyanin (PYO) is a major virulence factor secreted by Pseudomonas aeruginosa, and autophagy is a crucial homeostatic mechanism for the interaction between the pathogens and the host. It remains unknown whether PYO leads to autophagy in macrophages by regulating histone acetylation. The high mobility group nucleosomal binding domain 2 (HMGN2) has been reported to regulate the PYO-induced autophagy and oxidative stress in the epithelial cells; however, the underlying molecular mechanism has not been fully elucidated. In this study, PYO was found to induce autophagy in macrophages, and the mechanism might be correlated with the up-regulation of HMGN2 acetylation (HMGN2ac) and the down-regulation of H3K27 acetylation (H3K27ac) by modulation of the activities of acetyltransferases and deacetylases. Moreover, we further demonstrated that the up-regulated HMGN2ac enhances its recruitment to the Ulk1 promoter, while the down-regulation of H3K27ac reduces its recruitment to the Ulk1 promoter, thereby promoting or inhibiting the transcription of Ulk1. In conclusion, HMGN2ac and H3K27ac play regulatory roles in the PYO-induced autophagy in macrophages.

Correlation of periodontal diseases with intracranial aneurysm formation: novel predictive indicators.

BACKGROUND: We investigated whether periodontal diseases, specifically, periodontitis and gingivitis, could be risk factors of the incidence of intracranial aneurysms (IAs). METHODS: We performed a case-control study to compare the differences in the periodontal disease parameters of 281 cases that were divided into the IAs group and non-IAs group. All cases underwent complete radiographic examination for IAs and examination for periodontal health. RESULTS: Comparing with those in the non-IAs group, the cases in the IAs group were older (53.95 ± 8.56 vs 47.79 ± 12.33, p < 0.001) and had a higher incidence of hypertension (76 vs 34, p = 0.006). Univariate logistic regression analysis revealed that age (> 50 years) and hypertension were predictive risk factors of aneurysm formation (odds ratio [OR] 1.047, 95% confidence interval [95% CI] 1.022-1.073, p < 0.001 and OR 2.047, 95% CI 1.232-3.401, p = 0.006). In addition, univariate and multivariate logistic regression analyses showed that the parameters of periodontal diseases, including gingival index, plaque index, clinical attachment loss, and alveolar bone loss, were significantly associated with the occurrence of IAs (all p < 0.05). For further statistical investigation, the parameters of periodontal diseases were divided into four layers based on the quartered data. Poorer periodontal health condition (especially gingival index > 1.1 and plaque index > 1.5) had the correlation with IAs formation (p = 0.007 and p < 0.001). CONCLUSION: Severe gingivitis or periodontitis, combining with hypertension, is significantly associated with the incidence of IAs.

Animal Experiment of a Novel Neurointerventional Surgical Robotic System with Master-Slave Mode.

In order to inspect and improve the system performance of the neuro-interventional surgical robot and its effectiveness and safety in clinical applications, we conducted ten animal experiments using this robotic system. Cerebral angiography was performed on ten experimental animals, and various mechanical performance indicators, operating time, X-ray radiation dosage to the experimental animals and the experimenter, and arterial damage in the experimental animals were recorded when the robotic system completed cerebral angiography. The results show that the robotic system can successfully complete the cerebral angiography surgery, and the mechanical performance is up to standard. The operating time is almost the same as the physician's operating time. And the mean X-ray radiation dosage received by the experimental animals and experimenter was 0.893 Gy and 0.0859 mSv, respectively. There were no complications associated with damage to the vascular endothelium. The robotic system can basically complete the relevant assessment indicators, and its system performance, effectiveness, and safety in clinical applications meet the standards, basically meeting the requirements of clinical applications of neurointerventional surgery.

Brd4 inhibition ameliorates Pyocyanin-mediated macrophage dysfunction via transcriptional repression of reactive oxygen and nitrogen free radical pathways.

Macrophages play critical roles in the first-line immune defense against airway infections caused by Pseudomonas aeruginosa (PA). The redox-active phenazine-pyocyanin (PCN), as one of the most essential virulence factors, facilities PA-related infection via a wide spectrum of cellular oxidative damages. However, little is known for PCN cytotoxicity in macrophages. In this study, besides showing PCN-mediated reactive oxygen species (ROS) indeed involved in macrophage viability and function impairment, we at the first time demonstrated a novel role of reactive nitrogen species (RNS) pathway causing cellular damage in PCN-challenged macrophages. Using small molecule inhibitor JQ1 targeting Bromodomain and extra-terminal family proteins, we showed restrained iNOS-dependent nitric oxide (NO) production correlated with abolished Brd4 recruitment to the NOS2 (encoding inducible nitric oxide synthase-iNOS) promoter. Application of JQ1 diminished PCN-mediated peroxynitrite (ONOO-) that followed ROS and NO induction, restored macrophage survival and bacteria clearance as well as repressed local inflammation in PA/PCN-challenged mice lungs. Our results uncover a novel link between PCN-mediated macrophage dysfunction and reactive free radicals that rely on Brd4-dependent transcription modulation of multiple stress-response genes, suggesting Brd4 could be a promising therapeutic target in treating PA-related lung infection.

High-mobility group protein N2 induces autophagy by activating AMPK/ULK1 pathway and thereby boosts UPEC proliferation within bladder epithelial cells.

Urinary tract infection is one of the most common bacterial infections which is mainly caused by Escherichia coli (UPEC). Autophagy plays a key role in immune response to eliminate invading pathogens. Exploring the effect of autophagy on UPEC infection and the molecular mechanisms will be benefit for the treatment of urinary tract infection. High-mobility group protein N2 (HMGN2), a highly conserved nuclear protein and an antibacterial peptide, has been associated with bacterial infection induced immune response; however, whether this function is due to the regulation of autophagy remains unclear. In this study, we demonstrate for the first time that HMGN2 is upregulated in UPEC infection of bladder epithelial cell line 5637 (BEC 5637). Furthermore, HMGN2 enhances autophagy in BEC 5637 via activation of AMPK and ULK1, whereas UPEC suppresses autophagy. In addition, the enhanced autophagy activity by HMGN2 overexpression or rapamycin boosts the proliferation of UPEC J96 in BEC 5637. In summary, our data indicate that HMGN2 activates autophagy via AMPK/ULK1 pathway which can be utilized by UPEC J96 for their proliferation within bladder epithelial cells.

Euonymus alatus and its monomers alleviate liver fibrosis both in mice and LX2 cells by blocking TßR1-Smad2/3 and TNF-α-NF-κB pathways.

This study aimed to investigate the protective effects, effective constituents and preliminary mechanisms of Euonymus alatus on liver fibrosis and screen new high-efficacy drug for fibrosis. 112 male C57BL/6 mice were randomly divided into 14 groups: control group (CG), CCL4 group (CTG), low/medium/high dose of Euonymus alatus ethanol extracts (EAE), catechin (CA), dihydroquercetin (DHQ) and kaempferol (KA) groups. The study lasted for 30 days by injecting CCL4 in peritoneal cavity to make fibrosis model, all mice were sacrificed to observe morphological changes and collagenous fiber by HE and Masson staining, to test liver index, ALT, AST, to measure the expression of α-SMA and collagen I by immunohistochemistry and western blotting, to discuss the pathways of TßR1-Smad2/3 and TNF-α-NF-κB by WB and Elisa; after being evaluated the efficacy, anti-fibrosis drug of highest efficacy was chosen to repeat these indexes in human hepatic stellate cells-LX2. Results showed that EAE/CA/DHQ/KA prevented increases in liver index, ALT, AST, α-SMA, collagen I, TßR1, Smad2/3, TNF-α and p-NF-κB caused by CCL4 in dose-dependence, they also improved the liver morphology, decreased inflammatory cell infiltration and collagenous fiber in dose-dependence, CA' efficacy was best in mice; in LX-2, CA also decreased the expression of α-SMA, collagen I, TGF-ß, Smad2/3. All findings suggested that Euonymus alatus could alleviate liver inflammation and fibrosis by inhibiting TßR1-Smad2/3 and TNF-α-NF-κB pathways, flavonoid were effective constituents and catechin was screened as a new star for its best performance.

Initial Clinical Trial of Robot of Endovascular Treatment with Force Feedback and Cooperating of Catheter and Guidewire.

To evaluate the feasibility and safety of the robot of endovascular treatment (RobEnt) in clinical practice, we carried out a cerebral angiography using this robot system. We evaluated the performance of application of the robot system to clinical practice through using this robotic system to perform the digital subtraction angiography for a patient who was suspected of suffering intracranial aneurysm. At the same time, through comparing the postoperative head nuclear magnetic and blood routine with the preoperative examination, we evaluated the safety of application of the robot system to clinical practice. We performed the robot system to complete the bilateral carotid artery and bilateral vertebral arteriography. The results indicate that there was no obvious abnormality in the patient's cerebral artery. No obvious abnormality was observed in the examination of patients' check-up, head nuclear magnetism, and blood routine after the digital subtraction angiography. From this clinical trial, it can be observed that the robot system can perform the operation of cerebral angiography. The robot system can basically complete the related observation indexes, and its accuracy, effectiveness, stability, and safety basically meet the requirements of clinical application in neurointerventional surgery.

Operation evaluation in-human of a novel remote-controlled vascular interventional robot.

Remote-controlled vascular interventional robots (RVIRs) are being developed to increase the accuracy of surgical operations and reduce the number of occupational risks sustained by intervening physicians, such as radiation exposure and chronic neck/back pain. However, complex control of the RVIRs improves the doctor's operation difficulty and reduces the operation efficiency. Furthermore, incomplete sterilization of the RVIRs will increase the risk of infection, or even cause medical accidents. In this study, we introduced a novel method that provides higher operation efficiency than a previous prototype and allows for complete robot sterilization. A prototype was fabricated and validated through laboratory setting experiments and an in-human experiment. The results illustrated that the proposed RVIR has better performance compared with the previous prototype, and preliminarily demonstrated that the proposed RVIR has good safety and reliability and can be used in clinical surgeries.

Nuclear protein HMGN2 attenuates pyocyanin-induced oxidative stress via Nrf2 signaling and inhibits Pseudomonas aeruginosa internalization in A549 cells.

Pyocyanin (PCN, 1-hydroxy-5-methyl-phenazine) is one of the most essential virulence factors of Pseudomonas aeruginosa (PA) to cause various cytotoxic effects in long-term lung infectious diseases, however the early effect of this bacterial toxin during PA infection and subsequent autonomous immune response in host cells have not been fully understood yet. Our results display that early onset of PCN stimulates Pseudomonas aeruginosa PAO1 adhesion and invasion in A549 cells via ROS production. Non-histone nuclear protein HMGN2 is found to be involved in the regulation of PCN-induced oxidative stress by promoting intracellular ROS clearance. Mechanistically, HMGN2 facilitates nuclear translocation of transcription factor Nrf2 upon PCN stimulation and in turn elevates antioxidant gene expression. We also found that actin cytoskeleton dynamics is targeted by ROS, which is to be exploited by PAO1 for host cell internalization. HMGN2 regulates actin skeleton rearrangement in both PCN-dependent and independent manners and specifically attenuates PCN-mediated PAO1 infection via ROS elimination. These results uncover a novel link between nuclear protein HMGN2 and Nrf2-mediated cellular redox circumstance and suggest roles of HMGN2 in autonomous immune response to PA infection.

MiR-204 inhibits the proliferation and invasion of renal cell carcinoma by inhibiting RAB22A expression.

While miR-204 expression may be linked to renal cell carcinoma (RCC) progression, the detailed mechanisms remain unclear. In the present study, we demonstrated that miR-204 was differentially expressed in RCC tissues when compared with surrounding normal kidney tissues. Ectopic overexpression of miR-204 in human RCC cells suppressed cell proliferation and invasion in vitro and in vivo. Mechanism dissection revealed that miR-204 may function through RAB22A signals to inhibit RCC proliferation and invasion. Overexpression of RAB22A by oe-RAB22A was able to partially reverse the miR-204-mediated suppression of RCC tumor progression. Together, these results revealed that miR-204 suppressed RCC proliferation and invasion by directly targeting the RAB22A gene. Targeting newly identified RAB22A with miR-204 may aid in the suppression of RCC proliferation and invasion.

[Effect of Pongamia pinnata root flavonoids on the quality of ulcer healing and expression of EGF and TGF-alpha in the rat model of gastric ulcer induced by acetic acid].

OBJECTIVE: To observe the effects and mechanisms of Pongamia pinnata root flavonoids (PRF) on the experimental gastric ulcer induced by acetic acid and to study the mechanism of PRF on the quality of ulcer healing. METHODS: The models were established by acetic acid erosion, the quality of ulcer healing of PRF on the model of gastric ulcer were observed. The contents of epidermal growth factor (EGF) in serum were determined by radioimmunoassay. The expression of EGF and transforming growth factor-alpha (TGF-alpha) were detected by immunohistochemistry (SP). RESULTS: PRF significantly inhibited ulcerative formation induced by acetic acid (P < 0.05, P < 0.01). PRF could significantly increase the EGF and TGF-alpha (P < 0.05, P < 0.01) expression of para-ulcer mucosa tissue and improve the EGF contents in blood serum (P < 0.05, P < 0.01). CONCLUSION: PRF increases the contents of EGF in serum and the expression of EGF and TGF-alpha in the tissue around gastric ulcer which might be one of possible mechanisms that PRF improves quality of ulcer healing.

[Screening of effective fraction on experimental gastric ulcer from Pongamia pinnata roots].

OBJECTIVE: To study the effect of extract from Pongamia pinnata roots on experimental gastric ulcer and screen the effective fraction. METHOD: The models of gastric mucosa damage were induced by absolute alcohol in rats and reserpine in mice to observed the effect of ethyl alcohol extract from P. pinnata roots (PRE) and different parts on experimental gastric ulcer. RESULT: PRE, acetic ether extract and n-butanol extract could significantly inhibit the gastric mucosa damage induced by absolute alcohol in rats and reserpine in mice. In absolute alcohol models the gastric ulcer rates of inhibition were 86.4%, 85.4%, 11.5%, respectively. In reserpine models the gastric ulcer rates of inhibition were 37.8%, 33.8%, 19.7%, respectively. CONCLUSION: PRE, acetic ether extract and n-butanol extract could significantly inhibit the gastric mucosa damage induced by absolute alcohol in rats and reserpine in mice. Acetic ether extract from P. pinnata roots has the best effect on experimental gastric ulcer.