GLP-1 receptor agonist, liraglutide, protects podocytes from apoptosis in diabetic nephropathy by promoting white fat browning.

Glucagon like peptide-1 receptor agonists (GLP-1RAs) belong to the class of incretin drugs. Incretin is a hormone secreted into blood by intestinal cells after food stimulation that induces insulin secretion. Liraglutide is a long-acting GLP-1RA that can reduce blood pressure, blood lipids, and inflammation. Previous studies showed that liraglutide can promote white fat browning and improve renal outcomes in patients with type 2 diabetes mellitus. However, no studies have linked white fat browning to kidney damage. The objective of this study was to investigate the effects of liraglutide-induced white fat browning on podocyte apoptosis in diabetic nephropathy. We also aimed to determine whether podocytes express glucagon like peptide-1 receptor (GLP-1R) and if liraglutide directly affects podocytes via GLP-1R. We assessed fat and renal function in db/db and wild-type mice and the effects of adipocyte conditioned medium on cultured podocytes. Liraglutide (400 mg/kg/d) was subcutaneously injected for 8 weeks. Liraglutide promoted white fat browning in vivo. During adipogenic differentiation of 3T3-L1 cells in vitro, liraglutide also upregulated expression of peroxisome proliferator-activated receptor γ coactivator-1 alpha (PGC1α) and uncoupling protein 1 (UCP1), which can induce white fat browning in vitro. Furthermore, we found that supernatant from 3T3-L1 cells stimulated by liraglutide reduced podocyte apoptosis. The inhibitory effect of liraglutide on apoptosis was eliminated by exogenous TNF-α. Finally, podocytes express GLP-1R. In vivo and in vitro studies showed that the apoptosis of podocytes in diabetic nephropathy may be related to the effect of liraglutide on promoting white lipid browning. Similarly, liraglutide may directly affect podocytes via GLP-1R.

Analytical investigation of nano-bio interfacial protein mediation for fibroblast adhesion on hydroxyapatite nanoparticles.

A quartz crystal microbalance with dissipation (QCM-D) analysis was used to investigate fetal bovine serum (FBS) protein preadsorption on a hydroxyapatite (HAp) surface and the subsequent adhesion process of fibroblasts as compared with the case of oxidized poly(styrene) (PSox). The results showed that the preadsorption of FBS proteins on HAp promoted the subsequent initial cell adhesion ability. Moreover, the measured frequency (Δf) and dissipation shift (ΔD) curves, ΔD-Δf plots and viscoelastic analysis were used to study the initial cell adhesion process in real time. It was suggested that FBS-HAp showed sensitive changes in mass and viscoelasticity as compared with FBS-PSox, which realized the in situ reflection of the cell adhesion state, and the interfacial reactions between the cells and FBS-HAp surfaces such as dehydration and binding occurred to promote the initial cell adhesion and spreading. The viscoelastic analysis of the interface layer showed that the adhered cells on FBS-HAp could secrete some viscous substances such as extracellular matrix (ECM) proteins at the interfaces to provide good adhesion behaviors, and the Voigt-based viscoelastic model could clearly reveal the cellular interfacial viscoelasticity depending on the substrate surface. In addition, the morphology of cells was observed by confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM), and it was found that the pseudopodia were more uniformly stretched on FBS-HAp than on FBS-PSox. Furthermore, the state of the interfacial protein layer was analyzed by localized Fourier-transform infrared (FT-IR) spectroscopy and fluorescence microscopy (FLM), and it was indicated that the type of substrate affects the formation state of ECM proteins, resulting in changes in cell adhesion properties and morphology. The abundant formation of connective proteins (i.e., collagen type I) on FBS-HAp promoted subsequent pseudopodia formation and cell spreading. Therefore, the initial adhesion properties of fibroblasts on the FBS-HAp surface were systematically studied, which is of great importance for understanding the interfacial interaction between biomaterials and cells, and has great application value in biomedical fields.

Construction of an Emotional Lexicon of Patients With Breast Cancer: Development and Sentiment Analysis.

BACKGROUND: The innovative method of sentiment analysis based on an emotional lexicon shows prominent advantages in capturing emotional information, such as individual attitudes, experiences, and needs, which provides a new perspective and method for emotion recognition and management for patients with breast cancer (BC). However, at present, sentiment analysis in the field of BC is limited, and there is no emotional lexicon for this field. Therefore, it is necessary to construct an emotional lexicon that conforms to the characteristics of patients with BC so as to provide a new tool for accurate identification and analysis of the patients' emotions and a new method for their personalized emotion management. OBJECTIVE: This study aimed to construct an emotional lexicon of patients with BC. METHODS: Emotional words were obtained by merging the words in 2 general sentiment lexicons, the Chinese Linguistic Inquiry and Word Count (C-LIWC) and HowNet, and the words in text corpora acquired from patients with BC via Weibo, semistructured interviews, and expressive writing. The lexicon was constructed using manual annotation and classification under the guidance of Russell's valence-arousal space. Ekman's basic emotional categories, Lazarus' cognitive appraisal theory of emotion, and a qualitative text analysis based on the text corpora of patients with BC were combined to determine the fine-grained emotional categories of the lexicon we constructed. Precision, recall, and the F1-score were used to evaluate the lexicon's performance. RESULTS: The text corpora collected from patients in different stages of BC included 150 written materials, 17 interviews, and 6689 original posts and comments from Weibo, with a total of 1,923,593 Chinese characters. The emotional lexicon of patients with BC contained 9357 words and covered 8 fine-grained emotional categories: joy, anger, sadness, fear, disgust, surprise, somatic symptoms, and BC terminology. Experimental results showed that precision, recall, and the F1-score of positive emotional words were 98.42%, 99.73%, and 99.07%, respectively, and those of negative emotional words were 99.73%, 98.38%, and 99.05%, respectively, which all significantly outperformed the C-LIWC and HowNet. CONCLUSIONS: The emotional lexicon with fine-grained emotional categories conforms to the characteristics of patients with BC. Its performance related to identifying and classifying domain-specific emotional words in BC is better compared to the C-LIWC and HowNet. This lexicon not only provides a new tool for sentiment analysis in the field of BC but also provides a new perspective for recognizing the specific emotional state and needs of patients with BC and formulating tailored emotional management plans.

Angiogenesis-An Emerging Role in Organ Fibrosis.

In recent years, the study of lymphangiogenesis and fibrotic diseases has made considerable achievements, and accumulating evidence indicates that lymphangiogenesis plays a key role in the process of fibrosis in various organs. Although the effects of lymphangiogenesis on fibrosis disease have not been conclusively determined due to different disease models and pathological stages of organ fibrosis, its importance in the development of fibrosis is unquestionable. Therefore, we expounded on the characteristics of lymphangiogenesis in fibrotic diseases from the effects of lymphangiogenesis on fibrosis, the source of lymphatic endothelial cells (LECs), the mechanism of fibrosis-related lymphangiogenesis, and the therapeutic effect of intervening lymphangiogenesis on fibrosis. We found that expansion of LECs or lymphatic networks occurs through original endothelial cell budding or macrophage differentiation into LECs, and the vascular endothelial growth factor C (VEGFC)/vascular endothelial growth factor receptor (VEGFR3) pathway is central in fibrosis-related lymphangiogenesis. Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), as a receptor of LECs, is also involved in the regulation of lymphangiogenesis. Intervention with lymphangiogenesis improves fibrosis to some extent. In the complex organ fibrosis microenvironment, a variety of functional cells, inflammatory factors and chemokines synergistically or antagonistically form the complex network involved in fibrosis-related lymphangiogenesis and regulate the progression of fibrosis disease. Further clarifying the formation of a new fibrosis-related lymphangiogenesis network may potentially provide new strategies for the treatment of fibrosis disease.

Mitochondrial Dysfunction in Metabolic Dysfunction Fatty Liver Disease (MAFLD).

Nonalcoholic fatty liver disease (NAFLD) has become an increasingly common disease in Western countries and has become the major cause of liver cirrhosis or hepatocellular carcinoma (HCC) in addition to viral hepatitis in recent decades. Furthermore, studies have shown that NAFLD is inextricably linked to the development of extrahepatic diseases. However, there is currently no effective treatment to cure NAFLD. In addition, in 2020, NAFLD was renamed metabolic dysfunction fatty liver disease (MAFLD) to show that its pathogenesis is closely related to metabolic disorders. Recent studies have reported that the development of MAFLD is inextricably associated with mitochondrial dysfunction in hepatocytes and hepatic stellate cells (HSCs). Simultaneously, mitochondrial stress caused by structural and functional disorders stimulates the occurrence and accumulation of fat and lipo-toxicity in hepatocytes and HSCs. In addition, the interaction between mitochondrial dysfunction and the liver-gut axis has also become a new point during the development of MAFLD. In this review, we summarize the effects of several potential treatment strategies for MAFLD, including antioxidants, reagents, and intestinal microorganisms and metabolites.

Psychometric testing of the Chinese National Health Service Sustainability Model as an instrument to assess innovation in Chinese nursing settings.

OBJECTIVES: To conduct psychometric testing of the Chinese version of the National Health Service Sustainability Model as an instrument to assess the sustainability of innovation in the Chinese nursing setting. BACKGROUND: Evidence-based practice is recognized worldwide as a way to improve the quality of healthcare; however, many evidence-based practice programmes decline over time and do not sustain the benefits of their improvements. A sustainability assessment tool is used internationally but its use has not been validated in China. DESIGN: A methodological study to test instrument validity and reliability. METHODS: The data collection was conducted from 15 June 2022 to 31 August 2022. The internal consistency of the Chinese version of the sustainability model was measured with Cronbach's alpha. Confirmatory factor analysis was used to test the model's structural validity. RESULTS: Four hundred eighty-three questionnaires were returned, of which 478 were valid. The short time taken to evaluate the Chinese version of the sustainability model demonstrated its efficiency and ability to adapt to a busy clinical environment. The confirmatory factor analysis showed a good fit model and supported the convergence validity of the sustainability model. The Cronbach's alpha coefficient was 0.905 for the total scale, which indicated good internal consistency. CONCLUSIONS: The results of this study suggest that the Chinese version of the sustainability model is a valid, reliable and efficient tool for measuring the sustainability of evidence-based practices in Chinese nursing settings.

Investigation into self-assembled collagen arrays guided by the surface properties of polyimide films.

The mechanism of highly-oriented collagen (Col) fibril arrays on rubbed polyimide (PI) films was investigated in order to understand the interfacial Col-PI interactions. It was found that the orientation of the surface functional groups of the rubbed PI films was most effectively controlled and optimized by the rubbing conditions. In particular, nano-grooves with a width of 100-600 nm and a depth of 2-10 nm were formed on the rubbed PI films at a rubbing strength of 2.4 m, leading to the formation of the highest density of the Col fibril array. Moreover, highly-oriented Col fibrils were formed inside the nano-grooves by the formation of hydrogen bonds between the CO of the imide groups (@ rubbed PI films) and the N-H of the amino groups (@ ß-Sheets of Col molecules), resulting in the orientation of the Col molecules and subsequent assembly to the fibrils. Thus, the orientation and density of the fibril arrays on the rubbed PI films were successfully controlled by the interfacial interactions between the ß-Sheet component of Col and the nano-groove surfaces of the rubbed PI films. Therefore, the novel technology of this study will provide an effective method to fabricate the one-directional fibrous nanostructures and to understand how to control the orientation of biomolecules in vitro.

Perceptions of nurses and physicians on pay-for-performance in hospital: A systematic review of qualitative studies.

AIMS: This study aimed to systematically examine perceptions of nurses and physicians on pay-for-performance in hospital. BACKGROUND: Pay-for-performance projects have proliferated over the past two decades, most systematic reviews of which solely focused on its effectiveness in primary health care and the physicians' or nurses' attitudes. However, systematic reviews of qualitative approaches for better examining perceptions of both nurses and physicians in hospital were lacking. EVALUATION: Electronic databases were systematically searched with date from the inception to 31 December 2020. Meta-aggregation synthesis methodology and the conceptual framework of the theory of planned behaviour were used to summarize findings. KEY ISSUES: A total of nine studies were included. Three major synthesized themes were identified: (1) perceptions of the motivation effects and positive outcomes, (2) perceptions about the design defects and negative effects and (3) perceptions of the obstacles in the implementation process. CONCLUSION: To maximize the intended positive effects, nurses' and physicians' perceptions should be considered and incorporated into the project design and implementation stage. IMPLICATIONS FOR NURSING MANAGEMENT: The paper gives enlightenment to nurse managers on improving and advancing the cause of nurses when planning for or evaluating their institutions' policies on pay-for-performance in the future research.

[Comparison Between Different Mitochondrial Staining Methods in Cell and Tissue Samples].

OBJECTIVE: To compare the effects of mitochondria staining between specific mitochondrial fluorescent probes and anti-mitochondrial protein antibody in cell and tissue samples. METHODS: The HepG2 cells fixed by 4% paraformaldehyde were stained with MitoTracker Deep Red (100 nmol/L) or anti-Grp75 antibody (75 nmol/L or 100 nmol/L). The human healthy liver tissue samples fixed by 4% paraformaldehyde were stained with 150 nmol/L MitoTracker Deep Red or anti-Grp75 antibody. The above stained cell and tissue samples were observed using confocal microscopy. RESULTS: We found non-specific staining in HeLa cells and obscure mitochondrial image using MitoTracker Deep Red probes, while clear tubular and punctate distribution using anti-Grp75 antibody. In contrast, we observed more specific and better effects of MitoTracker Deep Red probes-stained liver tissue samples as compared to the antibody. CONCLUSION: To visualize mitochondria, the anti-Grp75 antibody staining worked better on cells and the MitoTracker Deep Red probes are more suitable for tissue samples.

[Expression and Significance of HIF-3α in Mitochondria].

OBJECTIVE: To investigate the mitochondrial translocation of hypoxia inducible factor-3α (HIF-3α) under normoxia and hypoxia and its physiological and pathological meanings. METHODS: ① After hypoxic (1%O2) or DMOG, CoCl2 treatments mimicking the hypoxic treatment, Western blot and immunofluorescence were used to examine the HIF-3α expression in mitochondria of HeLa and ACHN cells, respectively. ②The protease sensitivity experiment was used to explore the sub-organelle localization of HIF-3α in mitochondria. ③Western blot was used to examine mitochondrial HIF-3α in the normal mouse tissues and human liver carcinoma tissues. RESULTS: ① In HeLa and ACHN cells, HIF-3α translocated to mitochondria under normoxia and hypoxia, and its mitochondrial expression was higher under hypoxia; ②The protease sensitivity of HIF-3α was similar to proteins locating in the mitochondrial outer membrane; ③Mitochondrial HIF-3α expressed in multiple normal mouse tissues; The expression of mitochondrial HIF-3α was higher in human liver carcinoma tissues than the normal and adjacent tissues. CONCLUSIONS: HIF-3α translocated to mitochondrial outer membrane under both normoxia and hypoxia, and hypoxia could up-regulated HIF-3α mitochondrial translocation. Meanwhile, the phenomenon may be involved in the process of liver carcinoma.

Intrauterine infusion of platelet-rich plasma improves fibrosis by transforming growth factor beta 1/Smad pathway in a rat intrauterine adhesion model.

This study aims to elucidate the effects of Platelet-rich plasma (PRP) in fibrosis development in intrauterine adhesion (IUA), and the associated underlying mechanisms are also explored, which are expected to be a potential therapeutic scheme for IUA. In this research, PRP was obtained and prepared from the peripheral venous blood of rats. A rat model was induced by mechanical injury. Further, PRP was directly injected into the uterus for treatment. The appearance and shape of the uterus were assessed based on the tissues harvested. The fibrosis biomarker levels were analyzed. The transforming growth factor beta 1 (TGF-ß1) and Mothers against decapentaplegic homolog 7 (Smad7) levels, the phosphorylation of Smad2 (p-Smad2), and the phosphorylation of Smad3 (p-Smad3) were analyzed, and the molecular mechanism was investigated by rescue experiments. It was found that PRP improved the appearance and shape of the uterus in IUA and increased endometrial thickness and gland numbers. The administration of PRP resulted in a decrease in the expressions of fibrosis markers including collagen I, α-SMA, and fibronectin. Furthermore, PRP increased Smad7 levels and decreased TGF-ß1 levels, p-Smad2, and p-Smad3. Meanwhile, administration of TGF-ß1 activator reversed the therapeutic effects of PRP in IUA. Collectively, the intrauterine infusion of PRP can promote endometrial damage recovery and improve endometrial fibrosis via the TGF-ß1/Smad pathway. Hence, PRP can be a potential therapeutic strategy for IUA.

Risk factors and prognostic analysis of right ventricular dysfunction after lung resection for NSCLC.

Objectives: Lung cancer is the leading cause of cancer death, and 80-85% of all lung cancer cases are non-small cell lung cancer (NSCLC). Surgical resection is the standard treatment for early-stage NSCLC. However, lung resection, a surgical procedure, can result in complications and increased mortality. Recent studies have shown a significant correlation between complications after lung resection and right ventricular dysfunction. Methods: Transthoracic echocardiography-derived right ventricular-pulmonary artery coupling (RV-PAC) was utilized to assess right ventricular function in these patients. Multivariate logistic regression analysis was also conducted to assess risk factors independently associated with RV-PA uncoupling. The 3- and 5-year cumulative survival rates were estimated with Kaplan-Meier curves, and differences between groups were analyzed using the Mantel-Cox log-rank test. Results: RV-PA uncoupling was defined as a TAPSE/PASP value < 0.67 mm/mm Hg according to spline analysis. The results of multivariable logistic regression analysis indicated that diabetes is an independent risk factor for right ventricular dysfunction after lung resection in patients with NSCLC. Kaplan-Meier analysis revealed a significant decrease in the survival rate of patients with RV-PA uncoupling at both the 3-year follow-up (73% vs 40%, p < 0.001) and 5-year follow-up (64% vs 37%, p < 0.001). Conclusions: After lung resection for NSCLC, the patient's right ventricular function predicts prognosis. Patients with right ventricular dysfunction, particularly those with diabetes mellitus, have a worse prognosis. It is crucial to actively prevent and correct risk factors to reduce the mortality rate in these patients.

Over-expression of microRNA-494 up-regulates hypoxia-inducible factor-1 alpha expression via PI3K/Akt pathway and protects against hypoxia-induced apoptosis.

BACKGROUND: Hypoxia-inducible factor-1 alpha (HIF-1α) is one of the key regulators of hypoxia/ischemia. MicroRNA-494 (miR-494) had cardioprotective effects against ischemia/reperfusion (I/R)-induced injury, but its functional relationship with HIF-1α was unknown. This study was undertaken to determine if miR-494 was involved in the induction of HIF-1α. RESULTS: Quantitative RT-PCR showed that miR-494 was up-regulated to peak after 4 hours of hypoxia in human liver cell line L02. To investigate the role of miR-494, cells were transfected with miR-494 mimic or miR-negative control, followed by incubation under normoxia or hypoxia. Our results indicated that overexpression of miR-494 significantly induced the expression of p-Akt, HIF-1α and HO-1 determined by qRT-PCR and western blot under normoxia and hypoxia, compared to negative control (p < 0.05). While LY294002 treatment markedly abolished miR-494-inducing Akt activation, HIF-1α and HO-1 increase under both normoxic and hypoxic conditions (p < 0.05). Moreover, apoptosis detection using Annexin V indicated that overexpression of miR-494 significantly decreased hypoxia-induced apoptosis in L02 cells, compared to control (p < 0.05). MiR-494 overexpression also decreased caspase-3/7 activity by 1.27-fold under hypoxia in L02 cells. CONCLUSIONS: Overexpression of miR-494 upregulated HIF-1α expression through activating PI3K/Akt pathway under both normoxia and hypoxia, and had protective effects against hypoxia-induced apoptosis in L02 cells. Thus, these findings suggested that miR-494 might be a target of therapy for hepatic hypoxia/ischemia injury.

[Rapid identification of chemical constituents in Salvia chinensis by HPLC-TOF-MS].

It's established a high-performance liquid chromatography-time of flight mass spectrometry(HPLC-TOF-MS) method to analyze chemical constituents in Salvia chinensis. The separation was performed on a SHISEIDO MG C18 reverse phase column (3.0 mm x 100 mm, 3 microm). The mobile phase consisted of acetonitrile (A) and water (containing 0.1% formic acid, B) was used as gradient elute. The gradient of a phase, 10%-90% (0-33 min), 90% (33-40 min). The flow rate was 0.6 mL x min(-1). Post-column split ratio was 2:1. Temperature of column was 25 degrees C. Time-of-flight mass spectrometer (TOF-MS) and electrospray ion source (ESI) was applied for qualitative analysis under positive ion mode, and mass scan range was m/z 100-1 000. As a result,28 of the major chemical constituents of S. chinensis were identified by HPLC-TOF-MS. In this study, a rapid and efficient method for studying the chemical constituents in S. chinensis by HPLC-TOF-MS was established, which paves a way for the quality control and further studies of the herb in vivo.

Investigation of Surface Layers on Biological and Synthetic Hydroxyapatites Based on Bone Mineralization Process.

In this review, the current status of the influence of added ions (i.e., SiO44-, CO32-, etc.) and surface states (i.e., hydrated and non-apatite layers) on the biocompatibility nature of hydroxyapatite (HA, Ca10(PO4)6(OH)2) is discussed. It is well known that HA is a type of calcium phosphate with high biocompatibility that is present in biological hard tissues such as bones and enamel. This biomedical material has been extensively studied due to its osteogenic properties. The chemical composition and crystalline structure of HA change depending on the synthetic method and the addition of other ions, thereby affecting the surface properties related to biocompatibility. This review illustrates the structural and surface properties of HA substituted with ions such as silicate, carbonate, and other elemental ions. The importance of the surface characteristics of HA and its components, the hydration layers, and the non-apatite layers for the effective control of biomedical function, as well as their relationship at the interface to improve biocompatibility, has been highlighted. Since the interfacial properties will affect protein adsorption and cell adhesion, the analysis of their properties may provide ideas for effective bone formation and regeneration mechanisms.

Study on the pathogenesis of MiR-6324 regulating diarrheal irritable bowel syndrome and bioinformatics analysis.

Objective: To investigate the pathogenesis of IBS-D by bioinformatics analysis of the differential microRNAs in rat colon tissue and to analyze and predict the function of their target genes. Methods: Twenty male Wistar rats of SPF class were randomly divided into two groups, the model group was manipulated using the colorectal dilatation method + chronic restraint stress method to establish the IBS-D model; while the blank group stroked the perineum at the same frequency. Screening of differential miRNAs after High-throughput sequencing of rat colon tissue. GO and KEGG analysis of target genes using the DAVID website, further mapping using RStudio software; the STRING database and the Cytoscape software were used to obtain the protein interaction network (PPI) of the target genes as well as the core genes. Finally, qPCR was used to detect the expression of target genes in the colon tissue of two groups of rats. Results: After the screening, miR-6324 was obtained as the key of this study. The GO analysis of target genes of miR-6324 is mainly involved in protein phosphorylation, positive regulation of cell proliferation, and intracellular signal transduction; it affects a variety of cellular components such as cytoplasm, nucleus, and organelles on the intracellular surface; it is also involved in molecular functions such as protein binding, ATP binding, and DNA binding. KEGG analysis showed that the intersecting target genes were mainly enriched in cancer pathways, proteoglycans in cancer, neurotrophic signaling pathway, etc. The protein-protein interaction network screened out the core genes mainly Ube2k, Rnf41, Cblb, Nek2, Nde1, Cep131, Tgfb2, Qsox1, and Tmsb4x. The qPCR results showed that the expression of miR-6324 decreased in the model group, but the decrease was not significant. Conclusion: miR-6324 may be involved in the pathogenesis of IBS-D as a potential biological target and provide further ideas for research on the pathogenesis of the disease or treatment options.

Corrigendum: Study on the pathogenesis of MiR-6324 regulating diarrheal irritable bowel syndrome and bioinformatics analysis.

[This corrects the article DOI: 10.3389/fphar.2023.1044330.].

Presence of Rare Variants is Associated with Poorer Survival in Chinese Patients with Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder with phenotypic and genetic heterogeneity. Recent studies have suggested an oligogenic basis of ALS, in which the co-occurrence of two or more genetic variants has additive or synergistic deleterious effects. To assess the contribution of possible oligogenic inheritance, we profiled a panel of 43 relevant genes in 57 sporadic ALS (sALS) patients and eight familial ALS (fALS) patients from five pedigrees in east China. We filtered rare variants using the combination of the Exome Aggregation Consortium, the 1000 Genomes and the HuaBiao Project. We analyzed patients with multiple rare variants in 43 known ALS causative genes and the genotype-phenotype correlation. Overall, we detected 30 rare variants in 16 different genes and found that 16 of the sALS patients and all the fALS patients examined harbored at least one variant in the investigated genes, among which two sALS and four fALS patients harbored two or more variants. Of note, the sALS patients with one or more variants in ALS genes had worse survival than the patients with no variants. Typically, in one fALS pedigree with three variants, the family member with three variants (Superoxide dismutase 1 (SOD1) p.V48A,  Optineurin (OPTN) p.A433V and TANK binding kinase 1 (TBK1) p.R573H) exhibited much more severe disease phenotype than the member carrying one variant (TBK1 p.R573H). Our findings suggest that rare variants could exert a negative prognostic effect, thereby supporting the oligogenic inheritance of ALS.

Functional analysis of alloreactive memory CD4+ T cells derived from skin transplantation recipient and naïve CD4+ T cells derived from untreated mice.

BACKGROUND: Memory T cells (T(M)s) exhibit differential susceptibility to many immunomodulatory strategies that induce immunologic tolerance in naïve T cells, which are believed to be an important barrier to inhibiting rejection and inducing tolerance. As skin grafts are a common model for acquiring T(M)s, we evaluated function of T(M)s derived from skin grafts. We also assessed the modulatory effects on memory T cells function of the microRNAs miR-155 and miR-181a, which are involved in regulating cytokine secretion and TCR sensitivity to antigen, respectively. METHODS: Memory CD4(+) T cells derived from skin-graft recipient mice, and naïve CD4(+) T cells from untreated mice, were isolated by negative magnetic selection, and then stimulated with dendritic cells pulsed with donor-specific antigens. Effector function and regulating mechanisms were assessed. RESULTS: In contrast to naïve CD4(+) T cells, CD4(+) T(M)s stimulated with donor-specific antigen could quickly generate effector function in terms of proliferation and cytokine secretion; miR-155 and miR-181a levels in CD4(+) T(M)s rapidly increased during immune response compared to naïve CD4(+) T cells. CONCLUSION: Memory CD4(+) T cells derived from skin grafts could be used as an experimental tool for evaluating effects of different immune-modulating strategies on T(M)s. Levels of miR-155 and miR-181a up-regulated quickly in T(M)s, which could be an important mechanism by which T(M)s mediate immune responses rapidly.

Biomimetic Mineralization in External Layer of Decalcified Fish Scale.

The mineralization process of the osseous layer, which is highly calcified in vivo, was successfully imitated by the immersion process of the decalcified fish scales in simplified simulated body fluid (SSBF). An alkali treatment was used to modify the native collagen in the decalcified Tilapia fish scale. After the alkali treatment, the mineralization was facilitated in SSBF. The XRD patterns and SEM-EDS observation results demonstrated that the externally-mineralized layers by the immersion process were highly similar to the osseous layer containing lower-crystalline hydroxyapatite, suggesting that the simple biomimetic precipitation process was developed.