The bone mesenchymal stem cell-derived exosomal miR-146a-5p promotes diabetic wound healing in mice via macrophage M1/M2 polarization.

A diabetic wound is a refractory disease that afflicts patients globally. MicroRNA-146a-5p (miR-146a-5p) is reported to represent a potential therapeutic target for diabetic wounds. However, microRNA easily degrades in the wound microenvironment. This study extracted bone marrow mesenchymal stem cell (BMSC)-derived exosomes (EXO). Electroporation technology was used to load miR-146a-5p into EXO (labeled as EXO-miR-146a). The endothelial cells (human umbilical vein endothelial cells [HUVECs]) and macrophages were cocultured in transwell chambers in the presence of high glucose. Cell proliferation, migration, and angiogenesis were measured with cell counting kit 8, scratch, and tube forming assays, respectively. Flow cytometry was introduced to validate the biomarker of macrophages and BMSCs. The expression level of macrophage polarization-related proteins and tumor necrosis factor receptor-associated factor 6 (TRAF6) was assessed with western blotting analysis. The full-thickness skin wound model was developed to verify the in vitro results. EXO-miR-146a promoted the proliferation, migration, and angiogenesis of HUVECs in the hyperglycemic state by suppressing the TRAF6 expression in vitro. Additionally, EXO-miR-146a treatment facilitated M2 but inhibited M1 macrophage polarization. Furthermore, EXO-miR-146a enhances reepithelialization, angiogenesis, and M2 macrophage polarization, thereby accelerating diabetic wound healing in vivo. The EXO-miR-146a facilitated M2 macrophage polarization, proliferation, migration, and angiogenesis of HUVECs through TRAF6, thereby ameliorating intractable diabetic wound healing. These results established the basis for using EXO to deliver drugs and revealed mediators for diabetic wound treatment.

Chemical constituents of Viscum coloratum (Kom.) Nakai and their cytotoxic activities.

A new diarylheptanoid, (1 R,2S,3S,5S)-2,3-dihydroxy-3',3''-dimethoxy-4'-de-O-methylcentrolobine (1) and a new bisabolane-type sesquiterpenoid, (1 R,7S)-1,12,13-trihydroxybisabola-3,10-diene (2), together with nineteen known compounds (3-21) were isolated from the EtOH extract of the stems and branches of Viscum coloratum (Kom.) Nakai. Their structures were elucidated by extensive analysis of 1 D and 2 D NMR spectra and from the HRESIMS. All the compounds were evaluated for their cytotoxic activity against eight human tumor cell lines.

Inhibition of PLA2G4E/cPLA2 promotes survival of random skin flaps by alleviating Lysosomal membrane permeabilization-Induced necroptosis.

Necrosis that appears at the ischemic distal end of random-pattern skin flaps increases the pain and economic burden of patients. Necroptosis is thought to contribute to flap necrosis. Lysosomal membrane permeabilization (LMP) plays an indispensable role in the regulation of necroptosis. Nonetheless, the mechanisms by which lysosomal membranes become leaky and the relationship between necroptosis and lysosomes are still unclear in ischemic flaps. Based on Western blotting, immunofluorescence, enzyme-linked immunosorbent assay, and liquid chromatography-mass spectrometry (LC-MS) analysis results, we found that LMP was presented in the ischemic distal portion of random-pattern skin flaps, which leads to disruption of lysosomal function and macroautophagic/autophagic flux, increased necroptosis, and aggravated necrosis of the ischemic flaps. Moreover, bioinformatics analysis of the LC-MS results enabled us to focus on the role of PLA2G4E/cPLA2 (phospholipase A2, group IVE) in LMP of the ischemic flaps. In vivo inhibition of PLA2G4E with an adeno-associated virus vector attenuated LMP and necroptosis, and promoted flap survival. In addition, microRNA-seq helped us determine that Mir504-5p was differentially expressed in ischemic flaps. A string of in vitro and in vivo tests was employed to verify the inhibitory effect of Mir504-5p on PLA2G4E, LMP and necroptosis. Finally, we concluded that the inhibition of PLA2G4E by Mir504-5p reduced LMP-induced necroptosis, thereby promoting the survival of random-pattern skin flaps.Abbreviations: AAV: adeno-associated virus; ACTA2/α;-SMA: actin alpha 2, smooth muscle, aorta; ALOX15/12/15-LOX: arachidonate 15- lipoxygenase; c-CASP8: cleaved caspase; c-CASP3: cleaved caspase 3; CTSD: cathepsin D; CTSB: cathepsin B; CTSL: cathepsin L; DMECs: primary mouse dermal microvascular endothelial cells; ELISA: enzyme-linked immunosorbent assay; F-CHP: 5-FAM-conjugated collagen hybridizing peptide; FISH: fluorescence in situ hybridization; HUVECs: human umbilical vein endothelial cells; LAMP1: lysosomal-associated membrane protein 1; LAMP2: lysosomal-associated membrane protein 2; LC-MS: liquid chromatography-mass spectrometry; LDBF: laser doppler blood flow; LMP: lysosomal membrane permeabilization; LPE: lysophosphatidylethanolamine; LPC: lysophosphatidylcholine; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MLKL: mixed lineage kinase domain-like; NDI: N-dodecylimidazole; PECAM1/CD31: platelet/endothelial cell adhesion molecule 1; PLA2G4A/cPLA2: phospholipase A2, group IVA (cytosolic, calcium-dependent); PLA2G4E/cPLA2: phospholipase A2, group IVE; qPCR: quantitative real-time polymerase chain reaction; RIPK1: receptor (TNFRSF)-interacting serine-threonine kinase 1; RIPK3: receptor-interacting serine-threonine kinase 3; RISC: RNA-induced silencing complex; ROS: reactive oxygen species; shRNA: short hairpin RNA; SQSTM1: sequestosome 1; TBHP: tert-butyl hydroperoxide; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labelling.

A Gut-Specific LITAF-Like Gene in Antheraea pernyi (Lepidoptera: Saturniidae) Involved in the Immune Response to Three Pathogens.

Antheraea pernyi (Guérin-Méneville 1855) is an important resource for silk, food, and biohealth products; however, exogenous pathogens largely affect the commercial application potential of this species. Since the gut is a key organ for the digestion and absorption of nutrients as well as for immune defense, we used comparative transcriptome analysis to screen for a gut-specific molecular tool for further functional research in A. pernyi. In total, 3,331 differentially expressed genes (DEGs) were identified in the gut compared with all other pooled tissues of A. pernyi, including 1,463 upregulated genes in the gut. Among these, we further focused on a lipopolysaccharide-induced tumor necrosis factor-α factor (LITAF) gene because of its high gut-specific expression and the presence of a highly conserved SIMPLE-like domain, which is related to the immune response to pathogenic infections in many species. The cDNA sequence of ApLITAF was 447-bp long and contained a 243-bp open reading frame encoding an 80-amino acid protein. Immune challenge assays indicated that ApLITAF expression was significantly upregulated in the gut of A. pernyi naturally infected with nucleopolyhedrovirus (NPV) or fed leaves infected with the gram-negative bacterium Escherichia coli (Migula 1895) and the gram-positive bacterium Bacillus subtilis (Ehrenberg 1835). Cell transfection showed that ApLITAF localized to the lysosome. Collectively, these results suggested that ApLITAF played a role in the immune response of A. pernyi and could facilitate the future research and breeding application in this species.

miR-383-5p inhibits human malignant melanoma cells function via targeting CENPF.

Human malignant melanoma (MM), is a type of skin cancer with high morbidity and mortality. In this study, we investigated the role of miR-383-5p in human MM cells in vitro. miR-383-5p expression was downregulated in MM cell lines compared with the human normal melanocyte cell line, and miR-383-5p overexpression inhibited the proliferation, migration, and invasion of M14 and A375 cells. Furthermore, miR-383-5p was able to effectively bind to the 3'UTR of CENPF mRNA. miR-383-5p expression was negatively correlated with CENPF expression and miR-383-5p overexpression inhibited CENPF protein expression in M14 and A375 cells. The overexpression of CENPF could effectively rescue the inhibitory effect on proliferation and invasion caused by miR-383-5p. Additionally, using publicly available databases, we showed that CENPF expression was upregulated in human MM tissues and could predict the prognosis of MM. In conclusion, miR-383-5p acts as a tumor suppressor in human MM by targeting CENPF, suggesting CENPF as a potential therapeutic target for human MM.

Liraglutide, a TFEB-Mediated Autophagy Agonist, Promotes the Viability of Random-Pattern Skin Flaps.

Random skin flaps are commonly used in reconstruction surgery. However, distal necrosis of the skin flap remains a difficult problem in plastic surgery. Many studies have shown that activation of autophagy is an important means of maintaining cell homeostasis and can improve the survival rate of flaps. In the current study, we investigated whether liraglutide can promote the survival of random flaps by stimulating autophagy. Our results show that liraglutide can significantly improve flap viability, increase blood flow, and reduce tissue oedema. In addition, we demonstrated that liraglutide can stimulate angiogenesis and reduce pyroptosis and oxidative stress. Through immunohistochemistry analysis and Western blotting, we verified that liraglutide can enhance autophagy, while the 3-methylladenine- (3MA-) mediated inhibition of autophagy enhancement can significantly reduce the benefits of liraglutide described above. Mechanistically, we showed that the ability of liraglutide to enhance autophagy is mediated by the activation of transcription factor EB (TFEB) and its subsequent entry into the nucleus to activate autophagy genes, a phenomenon that may result from AMPK-MCOLN1-calcineurin signalling pathway activation. Taken together, our results show that liraglutide is an effective drug that can significantly improve the survival rate of random flaps by enhancing autophagy, inhibiting oxidative stress in tissues, reducing pyroptosis, and promoting angiogenesis, which may be due to the activation of TFEB via the AMPK-MCOLN1-calcineurin signalling pathway.

Before-after cohort study to assess the efficacy of fractional ablative carbon dioxide laser treatment of pediatric hand scars.

The purpose of this study is to investigate the efficacy of fractional ablative carbon dioxide laser (AFXL) surgery in patients with pediatric hand scars. This study enrolled hand scar patients who received treatment in our hospital between May 2018 and April 2019. Patients were assigned to undergo AFXL surgery based on their personal intents and condition, whereas the fractional laser was used for stiffness and abnormal texture. Outcomes were as follows: hand function was evaluated using the Michigan hand outcomes questionnaire; scar condition was evaluated using the Vancouver scar scale and UNC4P scar scale. Total 30 pediatric patients (mean age, 11.4 years) were eligible for the study and laser-treated scars were significantly improved in Michigan hand outcomes questionnaire from 52.30 ± 6.14 to 66.91 ± 6.43 (p < 0.001). Provider-rated Vancouver scar scale dropped from 8.80 ± 2.75 to 6.73 ± 2.52 (p < 0.001). Patient-reported UNC4P scar scale declined from7.07 ± 2.02 to 4.73 ± 1.31 (p < 0.001). AFXL surgery can significantly improve hand function and appearance of pediatric hand scars, suggesting its advantages over traditional methods of operative intervention.

Worldwide productivity in the hand and wrist literature: A bibliometric analysis of four highly cited subspecialty journals.

OBJECTIVE: Hand and wrist research has recently shown obvious progress. The quantity and quality of publications from different nations, however, have not been analyzed. In our study, we aimed to assess the characteristics of worldwide productivity in hand and wrist literature using highly cited subspecialty journals. METHODS: Literature search using the Web of Science database was conducted to identify hand and wrist articles in four highly cited subspecialty journals from 2005 to 2014. The number of articles, impact factors and citations were analyzed to evaluate the contributions of different countries. Publication activity was adjusted for the countries by population size. RESULTS: A total of 4268 publications were identified. The number of articles showed a significant increase of 2.10-fold between 2005 and 2014 (p = 0.0001). North America, West Europe, and East Asia were the most prolific areas. The majority of publications (92.03%) were from high-income countries, 7.97% from middle-income countries, and no publications from lower-income countries. The United States published the most articles (53.89%), followed by United Kingdom (6.51%), Japan (6.14%), Canada (3.70%), and China (3.37%). Articles originating from the United States showed the greatest number of total 5-year impact factors (5y-IF) (4059.56) and total citations (17,998). When normalized to population size, United States ranked the first (7.16), followed by Sweden (6.53), and Netherlands (5.72). However, Netherlands (1.893) had the highest mean 5y-IF, followed by Germany (1.884) and Australia (1.883). Sweden had the highest average citations per article (11.38), followed by Germany (9.63), and Australia (9.08). CONCLUSIONS: The number of publications of hand and wrist research shows a significant increase during the past 10 years. The United States is the most productive country in hand and wrist literature. However, some European countries and Australia may have higher quality of articles according to mean 5y-IF and mean citations per article.