Abstract Human periodontal ligament stem cells (hPDLSCs) are promising cells for dental and periodontal regeneration. Objective This study aimed to develop novel alginate-fibrin fibers that encapsulates hPDLSCs and metformin, to investigate the effect of metformin on the osteogenic differentiation of hPDLSCs, and to determine the regulatory role of the Shh/Gli1 signaling pathway in the metformin-induced osteogenic differentiation of hPDLSCs for the first time. Methodology CCK8 assay was used to evaluate hPDLSCs. Alkaline phosphatase (ALP) staining, alizarin red S staining, and the expression of osteogenic genes were evaluated. Metformin and hPDLSCs were encapsulated in alginate-fibrinogen solutions, which were injected to form alginate-fibrin fibers. The activation of Shh/Gli1 signaling pathway was examined using qRT-PCR and western blot. A mechanistic study was conducted by inhibiting the Shh/Gli1 pathway using GANT61. Results The administration of 50 μM metformin resulted in a significant upregulation of osteogenic gene expression in hPDLSCs by 1.4-fold compared to the osteogenic induction group (P < 0.01), including ALP and runt-related transcription factor-2 (RUNX2). Furthermore, metformin increased ALP activity by 1.7-fold and bone mineral nodule formation by 2.6-fold (P<0.001). We observed that hPDLSCs proliferated with the degradation of alginate-fibrin fibers, and metformin induced their differentiation into the osteogenic lineage. Metformin also promoted the osteogenic differentiation of hPDLSCs by upregulating the Shh/Gli1 signaling pathway by 3- to 6- fold compared to the osteogenic induction group (P<0.001). The osteogenic differentiation ability of hPDLSCs were decreased 1.3- to 1.6-fold when the Shh/Gli1 pathway was inhibited, according to ALP staining and alizarin red S staining (P<0.01). Conclusions Metformin enhanced the osteogenic differentiation of hPDLSCs via the Shh/Gli1 signaling pathway. Degradable alginate-fibrin hydrogel fibers encapsulating hPDLSCs and metformin have significant potential for use in dental and periodontal tissue engineering applications. Clinical Significance Alginate-fibrin fibers encapsulating hPDLSCs and metformin have a great potential for use in the treatment of maxillofacial bone defects caused by trauma, tumors, and tooth extraction. Additionally, they may facilitate the regeneration of periodontal tissue in patients with periodontitis.

Abstract As an indispensable and crucial part of the Moutai-flavored liquor production process, stacking fermentation facilitates the accumulation of various microbiota as microbial materials in fermented grains used for liquor making. The diversity of and dynamic changes in highly complex microbiota during this fermentation phase were investigated by characterizing microbial communities through high-throughput sequencing with an Illumina PE300 platform. Through community composition and differentially abundant analyses, 12 key bacteria and 12 key fungi were identified, which contributed to major variations in community succession. Bacillus and Saccharomyces were overrepresented in the bacterial and fungal communities, respectively. In addition, differentially abundant analyses showed that the key microbiota were mainly structured from the early period to the middle period of fermentation and functional populations showed a large accumulation in later periods. Further, endogenous factors showed changing trends, and their potential correlations and varied interactions with the key microbiota were explored through PLS-DA modeling. All the findings in the current study enhances knowledge of complex key microbiota in stacking fermentation and provides theoretical support for optimizing microbial structures through the regulation of endogenous factors and improving fermentative quality.

Abstract This study aims to explore the molecular mechanism of neuronal axonal lesions caused by high glucose, and to investigate the protective effect of Lycium barbarum polysaccharide (LBP) on neuronal axon damage with in vitro models. A rat model of type II diabetic encephalopathy was established, and the rats were treated with LBP for 10 weeks. Morris water maze experiment was used to detect spatial learning and memory ability. Neuronal axon lesions were measured by in vivo neuron tracing experiment. Immunohistochemistry and Western-blot technology were employed to detect protein localization and expression. Combined with in vitro primary neuron culture, total internal reflection fluorescence microscopy was used to detect the effects of high glucose and LBP on neuronal axon transport. PI3K inhibitors LY294002 and an activator of insulin signaling pathway were used to investigate the possible molecular mechanism. In vivo and in vitro experiments showed that in the hippocampus tissue of STZ-induced type II diabetic encephalopathy model rats, neuronal axons were damaged, accompanied by hyperphosphorylation of Tau protein, which subsequently resulted in axonal transport damage and cognition dysfunction. LBP significantly reduced peripheral blood glucose and serum insulin levels in type II diabetic encephalopathy model rats, thereby alleviating peripheral insulin resistance. At the same time, LBP can significantly improve the learning and memory impairment and brain neuron axon pathology in model rats. LY294002 treatment can effectively block the PI3K/Akt signaling pathway. The phosphorylation level of Tau protein in the LY294002+ LBP treatment group is higher than that of the LBP intervention group, while the insulin treatment can effectively reverse the blocking effect of LY294002 on LBPs. LBP can reduce the phosphorylation level of Tau protein by up-regulating the PI3K/Akt/GSK3β signaling pathway, with a protective effect on neuronal axon damage, thereby improving the cognitive function performance of type II diabetic encephalopathy model rats.

Abstract Purpose The research is intended for clarification of the efficacy as well as the underlying mechanism of GSK-3β inhibitors on the advancement of acute lung injuries in acute necrotizing pancreatitis (ANP) in rats. Methods Seventy-two rats were randomly divided into 6 groups: (1)ANP-vehicle; (2)ANP-TDZD-8;(3)ANP-SB216763;(4)Sham-vehicle;(5)Sham-TDZD-8;(6)Sham-SB216763; Blood biochemical test, histopathological examination and immunohistochemical analysis of rats pancreas and lung tissues were performed. The protein expression of GSK-3β, phospho-GSK-3β (Ser9), iNOS, ICAM-1, TNF-α, and IL-10 were detected in lung tissues by Western-blot. Results The outcomes revealed that the intervention of GSK-3β inhibitors alleviated the pathological damage of pancreas and lung (P<0.01), reduced serum amylase, lipase, hydrothorax and lung Wet-to-Dry Ratio, attenuated serum concentrations of IL-1β and IL-6 (P<0.01), inhibited the activation of NF-κB, and abated expression of iNOS, ICAM-1 and TNF-α protein, but up-regulated IL-10 expression in lung of ANP rats (P<0.01). The inflammatory response and various indicators in ANP-TDZD-8 groups were lower than those in ANP-SB216763 groups. Conclusions Inhibition of GSK-3β weakens acute lung injury related to ANP via the inhibitory function of NF-κB signaling pathway. Different kinds of GSK-3β inhibitors have different effects to ANP acute lung injury.

Reports remain insufficient on whether and how prostate-specific membrane antigen (PSMA) can influence in vivo osseous metastasis of prostate cancer (PCa). In the present study, the authors induced stable expression of PSMA in mouse PCa cell line RM-1. In vivo osseous metastasis was induced in 37 6-week-old female C57BL/6 mice weighing 22.45 ± 0.456 g. RM-1 cells were actively injected into the femoral bone cavity, leading to bilateral dissymmetry of bone density in the femoral bone. Tumor cells were also detected in bone tissue by pathological examination. The impact on bone density was demonstrated by the significant difference between animals injected with RM-PSMA cells (0.0738 ± 0.0185 g/cm²) and animals injected with RM-empty plasmid cells (0.0895 ± 0.0241 g/cm²). The lytic bone lesion of the RM-PSMA group (68.4%) was higher than that of the control group (27.8%). Immunohistochemistry showed that the expression of both vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) was distinctly higher in the RM-PSMA group than in the control group, while ELISA and Western blot assay indicated that VEGF and MMP-9 were higher in the RM-PSMA group compared to the control group (in vitro). Thus, the present study proposed and then confirmed for the first time that PSMA can promote in vivo osseous metastasis of PCa by increasing sclerotic destruction of PCa cells. Further analyses also suggested that PSMA functions positively on the invasive ability of RM-1 by increasing the expression of MMP-9 and VEGF by osseous metastases in vivo

Este artigo apresenta uma comparação de desempenho de vários métodos para o projeto de memórias associativas com Redes Neurais Celulares (RNCs). Embora tenha ocorrido um interesse crescente em tal tipo de aplicação de RNCs, não há uma comparação compreensível do desempenho entre os métodos propostos na literatura. Neste artigo, métodos existentes para projeto são revisados e o desempenho deles é medido por simulações numéricas. O teste T de Student é então utilizado para comparar os resultados obtidos.

This paper presents a performance comparison of various methods for associative memory design in Cellular Neural Networks (CNNs). Even though there have been an increasing interest in such kind of application for CNNs, there is no comprehensive comparison of the performance among the proposed methods in the literature. In this paper, existing design methods are reviewed and their corresponding performances are measured in numerical simulations. The T -Student test is then employed in order to compare the results obtained.