Bacterial composition and physicochemical characteristics of sorghum based on environmental factors in different regions of China

The fermentation process for Jiang-flavored baijiu using sorghum as the raw material involves a variety of microorganisms. However, the specific physicochemical characteristics of sorghum and microbial composition on its surface have not been fully elucidated. We aimed to perform a comprehensive comparative analysis of the variations in physicochemical properties and surface microflora in waxy sorghum samples from three prominent production regions in China (Renhuai, Jinsha, and Duyun). Multivariate statistical assessments were conducted that incorporated local soil and climate variables. The results showed that Cyanobacteria, unclassified bacteria, Proteobacteria, Firmicutes, and Bacteroidota were the dominant bacteria in these regions. These bacteria were associated with ethyl acetate, ethyl caprylate, ethyl lactate, and butyl groups, which synergistically produce flavorful compounds. The surface bacterial communities were affected by soil total phosphorus, altitude, diurnal temperature range, monthly mean temperature, precipitation, and effective accumulated temperature. The findings of this study provide a new perspective on microorganisms related to Jiang-flavored baijiu and can help establish a reference for the stability of liquor quality

Wnt/β-catenin pathway regulates cementogenic differentiation of adipose tissue-deprived stem cells in dental follicle cell-conditioned medium.

The formation and attachment of new cementum is crucial for periodontium regeneration. Tissue engineering is currently explored to achieve complete, reliable and reproducible regeneration of the periodontium. The capacity of multipotency and self-renewal makes adipose tissue-deprived stem cells (ADSCs) an excellent cell source for tissue regeneration and repair. After rat ADSCs were cultured in dental follicle cell-conditioned medium (DFC-CM) supplemented with DKK-1, an inhibitor of the Wnt pathway, followed by 7 days of induction, they exhibited several phenotypic characteristics of cementoblast lineages, as indicated by upregulated expression levels of CAP, ALP, BSP and OPN mRNA, and accelerated expression of BSP and CAP proteins. The Wnt/β-catenin signaling pathway controls differentiation of stem cells by regulating the expression of target genes. Cementoblasts share phenotypical features with osteoblasts. In this study, we demonstrated that culturing ADSCs in DFC-CM supplemented with DKK-1 results in inhibition of β-catenin nuclear translocation and down-regulates TCF-4 and LEF-1 mRNA expression levels. We also found that DKK-1 could promote cementogenic differentiation of ADSCs, which was evident by the up-regulation of CAP, ALP, BSP and OPN gene expressions. On the other hand, culturing ADSCs in DFC-CM supplemented with 100 ng/mL Wnt3a, which activates the Wnt/β-catenin pathway, abrogated this effect. Taken together, our study indicates that the Wnt/β-catenin signaling pathway plays an important role in regulating cementogenic differentiation of ADSCs cultured in DFC-CM. These results raise the possibility of using ADSCs for periodontal regeneration by modifying the Wnt/β-catenin pathway

DFC-CM condition effect Wnt/β-catenin signaling pathway in ADSCs.

<p>(<b>A</b>) Immunocytochemical staining showed that ADSCs cultured in basic medium or in DFC-CM condition expressed β-catenin. Scale bar represents 100 µm. (<b>B</b>) Beta-Catenin levels were examined by Western blot analysis and scanning densitometer. Beta-actin was used as internal control. (<b>C</b>). LEF-1and TCF-4 mRNA were subjected to real time-PCR analysis after cells were cultured in DFC-CM or basal medium (control) for 7 days. The expression levels were normalized to those of β-actin. The results represent mean values (SD) from three independent experiments performed in triplicates. *<i>P</i><0.05 vs. the control group.</p

Isolation and identification of ADSCs.

<p>(<b>A</b>) Representative images of colonies formed by ADSCs at low seeding density after 2 weeks in culture. (<b>B</b>). Flow cytometry analysis of the expression of cell surface markers related to mesenchymal (CD31, CD90, CD105, CD146 and STRO-1) or hematopoietic stem cells (CD34 and CD45). Cont: isotype control. (<b>C</b>) After ADSCs were cultured under osteogenic inductive conditions for 21 days, mineralized nodules were detected following alizarin red staining. ADSCs formed lipid clusters that stained positive for Oil Red O after 21 days of adipogenic induction. Scale bars represent 100 µm.</p

Regulation of cementogenic differentiation of ADSCs by Wnt/β-catenin signaling pathway.

<p>(<b>A</b>) Immunostaining of BSP and CAP in ADSCs cultured in basal medium or DFC-CM with or without Wnt3a (100 ng/mL)/DKK-1 (100 ng/mL) treatment for 7 d. Scale bar represents 100 µm. (<b>B–E</b>) Expression of CAP (<b>B</b>), ALP (<b>C</b>), BSP (<b>D</b>), and OPN (<b>E</b>) genes in ADSCs cultured in basal medium or in DFC-CM with Wnt3a (100 ng/mL)/DKK-1 (100 ng/mL) for 7 d. PBS was used as control condition. The expression levels were normalized to that of β-actin. The results represent mean values (SD) from three independent experiments performed in triplicates. *<i>P</i><0.05 vs. the PBS group.</p

Primer sequences.

<p>Primer sequences.</p

Wnt3a or DKK1 activate or inactivate the Wnt/β-catenin signaling pathway.

<p>(<b>A</b>) The expression of β-catenin as well as GSK3β were measured by Western blot in ADSCs grown in DFC-CM or in basal medium (control) with or without the presence of Wnt3a (100 ng/mL)/DKK-1 (100 ng/mL) for 7 d. Beta-actin was used as control for equal loading. (<b>B, C</b>) Expression of LEF-1 (<b>B</b>) and TCF-4 (<b>C</b>) genes in ADSCs cultured in basal medium or in DFC-CM with Wnt3a (100 ng/mL)/DKK-1 (100 ng/mL) for 7 d. PBS was used as control condition. The expression levels were normalized to that of β-actin. The results represent mean values (SD) from three independent experiments performed in triplicates. *<i>P</i><0.05 vs. the PBS group.</p

DFC-CM microenvironments induce ADSC differentiation into cementoblast-like cells.

<p>(<b>A</b>) Real-time PCR analysis of the expression of CAP, ALP, BSP and OPN in ADSCs grown in DFC-CM for 7 days. The expression levels were normalized to that of β-actin. The results represent mean values ( ± SD) from three independent experiments performed in triplicates. *<i>P</i><0.05 vs. the control group. (<b>B</b>) CAP, BSP, and OPN were measured by Western blot in ADSCs grown in basal medium or in cells cultured in DFC-CM for 7 d. GAPDH was used as control for equal loading. (<b>C</b>) Quantitative analysis of the OPN, BSP, ALP and CAP. *<i>P</i><0.05 vs. the control group.</p