Punicalagin promotes mincle-mediated phagocytosis of macrophages via the NF-κB and MAPK signaling pathways.

Punicalagin (PUN) is a polyphenol derived from the pomegranate peel. It has been reported to have many beneficial effects, including anti-inflammatory, anti-oxidant, and anti-proliferation. However, the role of PUN in macrophage phagocytosis is currently unknown. In this study, we found that pre-treatment with PUN significantly enhanced phagocytosis by macrophages in a time- and dose-dependent manner in vitro. Moreover, KEGG enrichment analysis by RNA-sequencing showed that differentially expressed genes following PUN treatment were significantly enriched in phagocyte-related receptors, such as the C-type lectin receptor signaling pathway. Among the C-type lectin receptor family, Mincle (Clec4e) significantly increased at the mRNA and protein level after PUN treatment, as shown by qRT-PCR and western blotting. Small interfering RNA (siRNA) mediated knockdown of Mincle in macrophages resulted in down regulation of phagocytosis. Furthermore, western blotting showed that PUN treatment enhanced the phosphorylation of nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) in macrophages at the early stage. Mincle-mediated phagocytosis by PUN was inhibited by PDTC (a NF-κB inhibitor) and SB203580 (a p38 MAPK inhibitor). In addition, PUN pre-treatment enhanced phagocytosis by peritoneal and alveolar macrophages in vivo. After intraperitoneal injection of Escherichia coli (E.coli), the bacterial load of peritoneal lavage fluid and peripheral blood in PUN pre-treated mice decreased significantly. Similarly, the number of bacteria in the lung tissue significantly reduced after intranasal administration of Pseudomonas aeruginosa (PAO1). Taken together, our results reveal that PUN enhances bacterial clearance in mice by activating the NF-κB and MAPK pathways and upregulating C-type lectin receptor expression to enhance phagocytosis by macrophages.

Molecular mechanisms of the antibacterial activity of polyimide fibers in a skin-wound model with Gram-positive and Gram-negative bacterial infection in vivo.

Recently, the need for antibacterial dressings has amplified because of the increase of traumatic injuries. However, there is still a lack of ideal, natural antibacterial dressings that show an efficient antibacterial property with no toxicity. Polyimide (PI) used as an implantable and flexible material has been recently reported as a mixture of particles showing more desirable antibacterial properties. However, we have identified a novel type of natural polyimide (PI) fiber that revealed antibacterial properties by itself for the first time. The PI fiber material is mainly composed of C, N, and O, and contains a small amount of Ca and Cl; the characteristic peaks of polyimide appear at 1774 cm-1, 1713 cm-1, 1370 cm-1, 1087 cm-1, and 722 cm-1. PI fibers displayed significant antibacterial activities against Escherichia coli (as a Gram-negative bacteria model) and methicillin-resistant Staphylococcus aureus (MRSA, as a Gram-positive bacteria model) according to the time-kill kinetics in vitro, and PI fibers damaged both bacterial cell walls directly. PI fibers efficiently ameliorated a local infection in vivo, inhibited the bacterial burden, decreased infiltrating macrophages, and accelerated wound healing in an E. coli- or MRSA-infected wound model. In conclusion, PI fibers used in the present study may act as potent antibacterial dressings protecting from MRSA or E. coli infections and as promising candidates for antimicrobial materials for trauma and surgical applications.

Transcriptomic analysis and laboratory experiments reveal potential critical genes and regulatory mechanisms in sepsis-associated acute kidney injury.

Background: Sepsis-associated acute kidney injury (SA-AKI) is one of the most frequent and serious complications of sepsis. However, the transcriptional regulatory network of the pathophysiological mechanism of the kidney has not been revealed. This study identified new mechanisms in SA-AKI using bioinformatics analyses and laboratory-based experiments. Methods: We performed transcriptomic profiling of mouse kidneys after cecal ligation and puncture (CLP) to mimic clinical sepsis. RNA from kidney samples from the CLP and control groups was isolated and analyzed using bulk messenger RNA (mRNA)-seq. Differentially expressed genes (DEGs) between the two groups were identified, and GO, KEGG and GSEA pathway enrichment analyses were performed. The protein-protein interaction (PPI) network of DEGs and hub genes was analyzed. The hub genes were verified using quantitative real-time polymerase chain reaction (qPCR) or Western blotting. The interaction network, targeted microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) of hub genes were predicted, and the critical miRNA-hub gene regulatory axis was verified using qPCR, Western blotting, malondialdehyde (MDA) determination and flow cytometry. Correlation analyses of N6-adenosine methylation (m6A) RNA methylation regulators and hub genes and m6A modification analysis were performed. Results: A total of 4,754 DEGs were identified between the two groups using high-throughput sequencing. The pathways in which DEGs were enriched included ferroptosis (the highest enrichment score), apoptosis, and the PI3K-Akt, NF-kappa B and IL-17 signaling pathways. Seven (Hmox1, Spp1, Socs3, Mapk14, Lcn2, Cxcl1 and Cxcl12) of the 15 hub genes were involved in the KEGG pathway. mmu-miR-7212-5p-Hmox1 was a key RNA regulatory axis in ferroptosis. m6A RNA methylation modifications were involved in SA-AKI. The correlation analyses showed the close interactions among the m6A RNA methylation regulators and important hub genes. Conclusions: The findings of this study provide new insights into the mechanism regulating the occurrence and progression of SA-AKI. The mmu-miR-7212-5p-Hmox1 axis in ferroptosis and m6A RNA methylation regulators may have potential clinical significance for the future treatment of SA-AKI. The datasets generated for this study can be found in the repository of the GEO database (Series number: GSE186822).

Methylsulfonylmethane protects against lethal dose MRSA-induced sepsis through promoting M2 macrophage polarization.

BACKGROUND: Multi-drug-resistant bacterial infections, which have become a global threat, lack effective treatments. The discoveries of non-antibiotics with different modes of antibacterial action, such as methylsulfonylmethane (MSM), are a promising new treatment for multi-drug-resistant pathogens. METHODS: We constructed a mouse peritonitis infection model to evaluate the effects of MSM against methicillin-resistant Staphylococcus aureus (MRSA) infection. The time-kill kinetics of MSM against MRSA and the effect of MSM on the integrity of bacterial cell membrane were measured. Viability effects of MSM on THP1 cells were performed by CCK-8 cytotoxicity assay. Systematic inflammatory factor levels of mice were detected using ELISA. The immune response of peritoneal macrophages during MRSA-infection was evaluated using RNA sequencing. Gene Ontology function, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses, and correlation analyses were applied to analysis RNA sequencing data. RT-qPCR, western blotting and flow cytometry were performed to analysis the gene and protein expression levels of macrophages. RESULTS: In in vitro experiments, MSM did not show significant killing effects on the growth of MRSA directly and did not destroy bacterial membrane integrity. MSM also displayed no significant effects on the proliferative capacity of THP1 cells. However, MSM treatment protected mice against a lethal dose MRSA-infection and decreased systemic inflammation. MSM upregulated metabolic pathway in peritoneal macrophages, especial glycolysis, during MRSA infection. MSM increased the expression of M2 markers (such as Arg1), promoted phosphorylation of STAT3 (which regulates M2 polarization), and decreased the expression of M1 markers in peritoneal macrophages. Additionally, MSM treatment increased the expression of H3K18 lactylation specific target genes, including Arg1. GNE-140, the LDHA-specific inhibitor of glycolysis, blocked the MSM-induced Arg1 expression in this disease model. CONCLUSIONS: MSM protects against MRSA infection through immunomodulation. MSM promotes the expression of Arg1 by lactate-H3K18la pathway to control macrophage to M2 polarization; it firstly provides therapeutic potential for drug-resistant infections and sepsis.

Water extract of Cayratia albifolia C.L.Li root relieves zymosan A-induced inflammation by restraining M1 macrophage polarization.

BACKGROUND: Cayratia albifolia C.L.Li (CAC) is a traditional Chinese herbal medicine used to treat inflammatory diseases. Our laboratory has firstly reported that the water extract from CAC relieved lipopolysaccharide (LPS)-induced inflammation, however stronger evidence is still needed to prove its anti-inflammatory effects and the mechanisms involved are also ambiguous. PURPOSE: This study sought to provide more evidence for the application of CAC in alleviating infectious inflammation and disclose novel pharmacological mechanisms. METHODS: Mice were injected with zymA into their paws or peritoneal cavities, and then treated with CAC. ELISA, immunofluorescence and flow cytometry were performed to detect the cytokines (IL-1ß, IL-6, TNF-α and IL-10) generation, the cell infiltration, and the CD86 or CD206 expression of macrophages. Then in vitro assays were performed on bone marrow-derived macrophages (BMDMs) and peritoneal macrophages (PMs) to detect their expression of iNOS, arg-1 and the cytokines above. On mechanisms, western blotting (WB), electrophoretic mobility shift assay (EMSA) and flow cytometry were carried out to measure NF-κB transcriptional activity, mitochondrial bioactivity and the mTORC1 activation when BMDMs were stimulated by zymA and treated with CAC. Finally, the chemical components consisted in the extract were analyzed by LC-MS. RESULTS: 200 mg/kg CAC clearly inhibited zymA induced mouse paw edema and reduced the contents of IL-1ß, IL-6 and TNF-α rather than IL-10 in local tissues. CAC also reduced CD86 but not CD206 in macrophages in situ. Through in vitro experiments, it was discovered that CAC reduced the protein and mRNA levels of IL-1ß, IL-6 and TNF-α, and also inhibited iNOS expression, but showed no influence on IL-10 and arg-1 in macrophages. We found CAC reduced NF-κB transcriptional activity, down-regulated mitochondrial membrane potential and ROS levels, and inhibited mTORC1 activity. Finally, we identified 15 major compounds in the extract, among which 4-guanidinobutyric acid and kynurenic acid were the most abundant. CONCLUSION: This study provides further evidence that CAC significantly reduces zymA induced infectious inflammation. In addition, this novel data revealed that CAC restrained M1 rather than promoting M2 macrophages polarization via multi-target inhibitory effects, based on its potentially active components.

Novel electromagnetic-based navigation for percutaneous transforaminal endoscopic lumbar decompression in patients with lumbar spinal stenosis reduces radiation exposure and enhances surgical efficiency compared to fluoroscopy: a randomized controlled trial.

BACKGROUND: Percutaneous transforaminal endoscopic lumbar decompression (PTELD) is an emerging surgical alternative for treating lumbar spinal stenosis (LSS). However, the foraminoplasty procedure often requires repeated fluoroscopy, and endoscopy just offers a local view. No studies have focused on decreasing radiation exposure with electromagnetic navigation assistance. This study introduces a novel electromagnetic-based navigation (EMN) endoscopic system for PTELD in patients with LSS and compares the results in navigation and fluoroscopy groups. METHODS: Eighty-eight patients with LSS were randomized into either a navigation (44 patients) or fluoroscopy group. Duration of surgery, cannula placement time, radiation dose, blood loss, intraoperative pain assessment, and postoperative hospitalization stay were evaluated. The clinical outcomes were evaluated using a visual analogue scale (VAS), the Oswestry Disability Index (ODI), 6-minute walk test, and modified Macnab criteria. RESULTS: Eighty-five patients were followed-up for at least 12 months. The duration of surgery and cannula placement time were significantly more efficient in the navigation group (P=0.03 and P<0.001). Intraoperative pain assessment showed significantly less pain in the navigation group (P=0.038). The radiation dose was significantly higher in the fluoroscopy group than the navigation group (P<0.001). The VAS scores for back (P<0.001) and leg (P<0.001) pain improved significantly in both groups after surgery, as did the ODI (P<0.001) scores. Improvements in walking ability and Macnab criteria assessments at the 12-month follow-up, assessed subjective by patient assessments did not differ between the two groups. CONCLUSIONS: The EMN system used in PTELD for patients with LSS compared to fluoroscopy enhances efficiency for foraminoplasty, reduces intraoperative pain and levels of radiation exposure. It results in outcomes comparable with results using fluoroscopy.

Comparison of Preliminary clinical outcomes between percutaneous endoscopic and minimally invasive transforaminal lumbar interbody fusion for lumbar degenerative diseases in a tertiary hospital: Is percutaneous endoscopic procedure superior to MIS-TLIF? A prospective cohort study.

BACKGROUND: Percutaneous endoscopic transforaminal lumbar interbody fusion (PETLIF) has been used in the treatment of lumbar degenerative diseases, as a novel minimally invasive technique. OBJECTIVES: To compare the surgical trauma and the medium-short term postoperative outcomes of PETLIF and traditional minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF). METHODS: From April to August of 2018, 75 patients with lumbar degenerative diseases received PETLIF (Group PE, 35 cases) or MIS-TLIF (Group MIS, 40 cases) were enrolled in the prospective cohort study. We recorded the serum creatine kinase (CK) and C-reactive protein (CRP), blood loss, visual analog scale (VAS), Oswestry Disability Index (ODI), modified Macnab criteria score, complications, and fusion rates of the 2 groups. RESULTS: There were significant reductions in CRP (P = 0.002) on postoperative day (POD) 3, and CK (P = 0.011) on POD 1 for Group PE than Group MIS. The mean true total blood loss (P < 0.001), intraoperative blood loss (P < 0.001), postoperative drains (P < 0.001), and hidden blood (P = 0.020) in the Group PE were significantly less compared with Group MIS. The VAS score for low-back pain, leg pain and ODI score improved significantly in both groups after surgery (P < 0.05). The VAS of low-back pain on POD 1 was significant less (P < 0.001) for Group PE. There was no statistical difference (P = 0.561) in CT fusion rates between Group PE (85%) and Group MIS (92%). No serious complication was observed in any patients. CONCLUSION: The study indicated that PETLIF had advantages of less surgical trauma, less postoperative low-back pain, less hidden blood loss, and faster recovery, compared with MIS-TLIF. There was no significant difference in medium-short term surgical outcomes between the 2 techniques. However, the indications of PETLIF is relatively limited, and the learning curve of PETLIF is deep, surgeons need to select indications strictly. Further study with big sample size and long-term follow-up is needed.

Percutaneous Endoscopic Lumbar Discectomy Assisted by O-Arm-Based Navigation Improves the Learning Curve.

OBJECTIVE: There is a steep learning curve with traditional percutaneous endoscopic lumbar discectomy (PELD). The aim of this study is to assess the safety and efficacy of PELD assisted by O-arm-based navigation for treating lumbar disc herniation (LDH). METHODS: From September of 2017 to January of 2018, 118 patients with symptomatic LDH were enrolled in the prospective cohort study. The patients undergoing PELD with O-arm-based navigation technique were defined as group A (58 cases), and those undergoing traditional X-ray fluoroscopy method were defined as group B (60 cases). We recorded the operation time, cannula placement time, radiation exposure time, visual analog scale (VAS), Oswestry Disability Index (ODI), and Macnab criteria score of the 2 groups. RESULTS: The average operation time (95.21 ± 19.05 mins) and the cannula placement time (36.38 ± 14.67 mins) in group A were significantly reduced compared with group B (operation time, 113.83 ± 22.01 mins, P<0.001; cannula placement time, 52.63 ± 17.94 mins, P<0.001). The learning curve of PELD in group A was steeper than that in group B and was lower in the relatively flat region of the end. There were significant differences of the clinical parameters at different time points (VAS of low back, P < 0.001; VAS of leg, P < 0.001; and ODI, P < 0.001). The VAS scores for low back pain and leg pain improved significantly in both groups after surgery and gradually improved as time went by. No serious complication was observed in any patients in either group. CONCLUSION: The study indicated that PELD assisted by O-arm navigation is safe, accurate, and efficient for the treatment of lumbar intervertebral disc herniation. It reshaped the learning curve of PELD, reduced the difficulty of surgery, and minimized radiation exposure to surgeons. This study was registered at Chinese Clinical Trail Registry (Registration Number: ChiCTR1800019586).

Percutaneous Endoscopic Lumbar Interbody Fusion: Technical Note and Preliminary Clinical Experience with 2-Year Follow-Up.

OBJECTIVE: Endoscopic surgeries have been attempted in the field of lumbar decompression and fusion surgery in the past decade. Percutaneous endoscopic lumbar interbody fusion (PELIF) is a new-emerging technique taking advantages of an anatomical (Kambin's triangle) to achieve simultaneous decompression and fusion under endoscopic visualization. The purpose of this study is to evaluate the feasibility and safety of PELIF technique with general anesthesia and neuromonitoring. METHODS: The authors present the details of PELIF technique with general anesthesia and neuromonitoring. The first 7 consecutive patients treated with minimum of 2 year's follow-up were included. Clinical outcomes were assessed by visual analog scale (VAS) for back and leg pain, Oswestry Disability Index (ODI) scores, and the Short Form-36 health survey questionnaire (SF-36) in the immediate preoperative period and during the follow-up period. RESULTS: All patients underwent single-level PELIF surgery successfully and without conversion to open surgery. The average age was 56.0±13.0 years. All patients had Grade I degenerative/isthmic spondylolisthesis and 4 patients coexisted with disc herniation. The mean operative time was 167.5±30.9 minutes, and intraoperative blood loss was 70.0±24.5 ml. Postoperative drainage volume was 24.5±18.3 ml. The differences in the VAS scores for low back pain and leg pain between preoperative and follow-up were significant (P<0.05). The SF-36 Physical Component Summary (PCS) improved from 38.83±4.17 to 55.67±2.58 (P<0.001). The SF-36 Mental Component Summary (MCS) improved from 43.83±3.13 to 57.50±5.36 (P=0.001). The ODI score improvement rate was 33.7±3.7 %. All cases demonstrated radiopaque graft in the intervertebral disc space consistent with solid arthrodesis. CONCLUSIONS: PELIF technique seems to be a promising surgical technique for selected appropriate patients, with the minimal invasive advantages in decreased blood, shortage of ambulation time, and hospital stay, compared with MIS-TLIF. Because of limited Kambin's triangle space and the exiting nerve root nearby, PELIF is still a challenging technique. Future advancement and development in instrument and cage design are vital for application and popularization of this technique. Prospective, randomized, controlled studies with large sample size on PELIF technique are still needed to prove its safety, efficacy, and minimal invasive advantages.

A Novel Targeted Foraminoplasty Device Improves the Efficacy and Safety of Foraminoplasty in Percutaneous Endoscopic Lumbar Discectomy: Preliminary Clinical Application of 70 Cases.

OBJECTIVE: Percutaneous endoscopic lumbar discectomy (PELD) has become a mature and mainstream surgical technique for treating lumbar disc herniation (LDH); however, there is a steep learning curve with PELD, especially for puncture and foraminoplasty. In this study, we assessed the outcome and safety of a novel quantificational and targeted foraminoplasty device named ZESSYS for LDH. METHODS: From September to December of 2016, 70 patients with symptomatic LDH were enrolled in the study. The patients were assigned randomly to either the ZESSYS group or the conventional TESSYS group. We recorded the cannula introduction time, decompression time, radiation exposure time, intraoperative pain feeling score, visual analog scale, Oswestry Disability Index, and Macnab criteria score of the 2 groups. The mean follow-up period was 14 months. RESULTS: The average cannula introduction time (16.50 ± 3.29 minutes) and the radiation exposure time (40.71 ± 6.23 seconds) in the ZESSYS group were significantly reduced compared with the TESSYS group (cannula introduction time, 20.06 ± 3.37 minutes, P = 0.000; radiation exposure time, 49.20 ± 7.84 seconds, P = 0.000). Intraoperative pain feeling score in the ZESSYS group was significantly improved compared with the TESSYS group (P = 0.021). There were no significant differences between the 2 groups for decompression time (P = 0.617), Macnab criteria (P = 0.769), or visual analog scale and Oswestry Disability Index scores at the same time point (P > 0.05). No serious complication was observed in any patients in either group. CONCLUSIONS: The novel targeted foraminoplasty technique with the specially designed double-cannulas is an effective and safe treatment for lumbar intervertebral disc herniation. It reduces the difficulty of PELD learning, minimizes radiation exposure, and decreases intraoperative pain associated with foraminoplasty.