Identification and validation of NAD+ metabolism-related biomarkers in patients with diabetic peripheral neuropathy.

Background: The mechanism of Nicotinamide Adenine Dinucleotide (NAD+) metabolism-related genes (NMRGs) in diabetic peripheral neuropathy (DPN) is unclear. This study aimed to find new NMRGs biomarkers in DPN. Methods: DPN related datasets GSE95849 and GSE185011 were acquired from the Gene Expression Omnibus (GEO) database. 51 NMRGs were collected from a previous article. To explore NMRGs expression in DPN and control samples, differential expression analysis was completed in GSE95849 to obtain differentially expressed genes (DEGs), and the intersection of DEGs and NMRGs was regarded as DE-NMRGs. Next, a protein-protein interaction (PPI) network based on DE-NMRGs was constructed and biomarkers were screened by eight algorithms. Additionally, Gene Set Enrichment Analysis (GSEA) enrichment analysis was completed, biomarker-based column line graphs were constructed, lncRNA-miRNA-mRNA and competing endogenouse (ce) RNA networks were constructed, and drug prediction was completed. Finally, biomarkers expression validation was completed in GSE95849 and GSE185011. Results: 5217 DEGs were obtained from GSE95849 and 21 overlapping genes of DEGs and NMRGs were DE-NMRGs. Functional enrichment analysis revealed that DE-NMRGs were associated with glycosyl compound metabolic process. The PPI network contained 93 protein-interaction pairs and 21 nodes, with strong interactions between NMNAT1 and NAMPT, NADK and NMNAT3, ENPP3 and NUDT12 as biomarkers based on 8 algorithms. Expression validation suggested that ENPP3 and NUDT12 were upregulated in DPN samples (P < 0.05). Moreover, an alignment diagram with good diagnostic efficacy based on ENPP3 and NUDT12 were identified was constructed. GSEA suggested that ENPP3 was enriched in Toll like receptor (TLR) pathway, NUDT12 was enriched in maturity onset diabetes of the young and insulin pathway. Furthermore, 18 potential miRNAs and 36 Transcription factors (TFs) were predicted and the miRNA-mRNA-TF networks were constructed, suggesting that ENPP3 might regulate hsa-miR-34a-5p by affecting MYNN. The ceRNA network suggested that XLOC_013024 might regulate hsa-let-7b-5p by affecting NUDT12. 15 drugs were predicted, with 8 drugs affecting NUDT12 such as resveratrol, and 13 drugs affecting ENPP3 such as troglitazone. Conclusion: ENPP3 and NUDT12 might play key roles in DPN, which provides reference for further research on DPN.

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.

Synergistic Effect of Yak Dung Fiber and Yak Dung Ash on the Mechanical and Shrinkage Properties of Cement Mortar.

The high value use of agricultural and livestock waste resources in the context of a low carbon economy is a challenge that currently plagues many countries. Yak dung, as a waste resource from livestock farming in the plateau, is considered to be a misplaced treasure. In this work, yak dung was processed into yak dung fiber (YDF) and yak dung ash (YDA), respectively, and the microscopic morphology of the YDF and YDA was assessed using scanning electron microscopy (SEM). The elements in the YDA were analyzed by energy dispersive X-ray spectroscopy (EDX). Moreover, cementitious composites were prepared with YDF at 0%, 0.3%, 0.5% and 0.7% and by replacing cement with YDA at 5%, 10% and 15% to assess the workability, mechanical properties and shrinkage properties of cementitious composites containing different YDF types (alkali treated and untreated), contents and different YDA contents. The results showed that alkali-treated YDF and YDA contain a large number of honeycomb structure pores, and the strength of cementitious materials with alkali-treated YDF was higher. The addition of YDF had a negative effect on the fluidity and compressive strength of the cementitious material, but the 0.3% YDF was beneficial in increasing its flexural strength. The compressive strength and flexural strength first increased and then decreased with the increase of YDA content. Both YDF and YDA inhibited the shrinkage of the cement paste, and the shrinkage strain of the cement matrix composites containing 0.3% YDF and 10% YDA was reduced by 51.1% compared to the control group. This work is expected to promote the application of yak dung waste in cementitious materials.

Effects of Ambient Humidity on Water Migration and Hydrate Change in Early-Age Hardened Cement Paste.

Ultra-low humidity environments will lead to changes in the microstructure of C-S-H, which will reduce the mechanical properties and service life of cement-based concrete. Thus, to further explore the mechanism on the microscale, this paper studied the water migration and the changes in the hydration products in white cement that was cured for 7 days at 20 °C and at different ambient relative humidities (RHs). The migration and transformation of different types of water in cement paste were studied by low-field nuclear magnetic resonance (NMR). At the same time, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to analyze semi-quantitatively the crystal phase in the hydration products. The results showed that in the first 7 days of the curing process, the content of the different types of water and the hydration products in the cement samples were influenced by the ambient RH. The total water content of the samples will decrease with the decrease in the RH; when the RH decreases to 54% or below, the chemically bound water in the samples will increase with the decline in the RH. Additionally, when the ambient RH is lower than 54%, the grossular will gradually transform into hydrogrossular crystals with the decrease in the RH, and the hibschite with less chemically bound water will transform into katoite with more chemically bound water. In future research, the water migration and hydrate changes under different curing ages, drying processes, and coupling effects should be explored.

Pupal Diapause Termination and Transcriptional Response of Antheraea pernyi (Lepidoptera: Saturniidae) Triggered by 20-Hydroxyecdysone.

The pupal diapause of univoltine Antheraea pernyi hampers sericultural and biotechnological applications, which requires a high eclosion incidence after artificial diapause termination to ensure production of enough eggs. The effect of pupal diapause termination using 20-hydroxyecdysone (20E) on the eclosion incidence has not been well-documented in A. pernyi. Here, the dosage of injected 20E was optimized to efficiently terminate pupal diapause of A. pernyi, showing that inappropriate dosage of 20E can cause pupal lethality and a low eclosion incidence. The optimal ratio of 20E to 1-month-old pupae was determined as 6 µg/g. Morphological changes showed visible tissue dissociation at 3 days post-injection (dpi) and eye pigmentation at 5 dpi. Comprehensive transcriptome analysis identified 1,355/1,592, 494/203, 584/297, and 1,238/1,404 upregulated and downregulated genes at 1, 3, 6, and 9 dpi, respectively. The 117 genes enriched in the information processing pathways of "signal transduction" and "signaling molecules and interaction" were upregulated at 1 and 3 dpi, including the genes involved in FOXO signaling pathway. One chitinase, three trehalase, and five cathepsin genes related to energy metabolism and tissue dissociation showed high expression levels at the early stage, which were different from the upregulated expression of four other chitinase genes at the later stage. Simultaneously, the expression of several genes involved in molting hormone biosynthesis was also activated between 1 and 3 dpi. qRT-PCR further verified the expression patterns of two ecdysone receptor genes (EcRB1 and USP) and four downstream response genes (E93, Br-C, ßFTZ-F1, and cathepsin L) at the pupal and pharate stages, respectively. Taken together, these genes serve as a resource for unraveling the mechanism underlying pupal-adult transition; these findings facilitate rearing of larvae more than once a year and biotechnological development through efficient termination of pupal diapause in A. pernyi in approximately half a month.

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.

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.

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.

Targeting TFE3 Protects Against Lysosomal Malfunction-Induced Pyroptosis in Random Skin Flaps via ROS Elimination.

Increasing evidence indicates that pyroptosis, a new type of programmed cell death, may participate in random flap necrosis and play an important role. ROS-induced lysosome malfunction is an important inducement of pyroptosis. Transcription factor E3 (TFE3) exerts a decisive effect in oxidative metabolism and lysosomal homeostasis. We explored the effect of pyroptosis in random flap necrosis and discussed the effect of TFE3 in modulating pyroptosis. Histological analysis via hematoxylin-eosin staining, immunohistochemistry, general evaluation of flaps, evaluation of tissue edema, and laser Doppler blood flow were employed to determine the survival of the skin flaps. Western blotting, immunofluorescence, and enzyme-linked immunosorbent assays were used to calculate the expressions of pyroptosis, oxidative stress, lysosome function, and the AMPK-MCOLN1 signaling pathway. In cell experiments, HUVEC cells were utilized to ensure the relationship between TFE3, reactive oxygen species (ROS)-induced lysosome malfunction and cell pyroptosis. Our results indicate that pyroptosis exists in the random skin flap model and oxygen and glucose deprivation/reperfusion cell model. In addition, NLRP3-mediated pyroptosis leads to necrosis of the flaps. Moreover, we also found that ischemic flaps can augment the accumulation of ROS, thereby inducing lysosomal malfunction and finally initiating pyroptosis. Meanwhile, we observed that TFE3 levels are interrelated with ROS levels, and overexpression and low expression of TFE3 levels can, respectively, inhibit and promote ROS-induced lysosomal dysfunction and pyroptosis during in vivo and in vitro experiments. In conclusion, we found the activation of TFE3 in random flaps is partially regulated by the AMPK-MCOLN1 signal pathway. Taken together, TFE3 is a key regulator of ROS-induced pyroptosis in random skin flaps, and TFE3 may be a promising therapeutic target for improving random flap survival.

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.

Comparison of bacterial communities between midgut and midgut contents in two silkworms, Antheraea pernyi and Bombyx mori.

Bacterial communities living inside the midgut of insects have been attracting increasing interest. Previous studies have shown that both the midgut and midgut contents harbor bacterial communities. However, whether the bacterial communities of the insect midgut are similar to those of the insect midgut contents (including the peritrophic membrane, food particles, and digestive fluids secreted by the midgut in this study) remains unknown. In the present study, we analyzed two economically important silkworms, the Chinese oak silkworm Antheraea pernyi (Lepidoptera: Saturniidae) and the mulberry silkworm Bombyx mori (Lepidoptera: Bombycidae), through Illumina MiSeq technology to address this issue. In A. pernyi larvae, 17 phyla and 162 genera were found in the midgut, while 7 phyla and 36 genera were found in the midgut contents. For B. mori larvae, 30 phyla and 465 genera were found in the midgut, but 22 phyla and 344 genera were found in the midgut contents. This evidence from the two silkworms suggests that the bacterial composition and diversity in the midgut are more diverse than those in the midgut contents. Principal component analysis revealed a significant difference in the bacterial community structure between the midgut and midgut contents of B. mori. To our knowledge, this is the first study to compare the bacterial communities between the midgut and midgut contents in insects, and the results will provide useful information for probing the functional differentiation within the midgut in the future.

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.

The complete mitochondrial genome of the Chinese oak silkmoth, Antheraea pernyi (Lepidoptera: Saturniidae).

We determined the complete nucleotide sequence of the mitogenome from Chinese oak silkmoth, Antheraea pernyi (Lepidoptera: Saturniidae). The 15,566 bp circular genome contains a typical gene organization and order for lepidopteran mitogenomes. The mitogenome contains the lowest A+T content (80.16%) among the known lepidopteran mitogenome sequences. An unusual feature is the occurrence of more Ts than As, with a slightly negative AT skewness (-0.021), in the composition of the major genome strand. All protein-coding genes are initiated by ATN codons, except for cytochrome oxidase subunit I, which is proposed by the TTAG sequence as observed in other lepidopterans. All transfer RNAs (tRNAs) have a typical clover-leaf structure of mitochondrial tRNA, except for tRNASer(AGN), the DHU arm of which could not form a stable stem-loop structure. Two aligned sequence blocks with a length of more than 50 bp and 90% of the sequence identity were identified in the A+T-rich region of the Saturniidae and Bombycoidae species.