[Latest Research Findings on Immune Microenvironment Regulation in Jawbone-Related Diseases].

Among the bones of the whole body, the jawbone is considered the region where remodeling takes place the most actively. In this region, the homeostasis of the bones is maintained by the balanced activities of osteoblasts and osteoclasts. The bone immune microenvironment can simultaneously regulate osteoblasts and osteoclasts. Compared with other bones, the jawbone is more susceptible to pathogens because it is exposed to the bacteria in the oral environment. In the case of inflammatory pathology, an over-activated immune system stimulates the activation of osteoclasts and inhibits osteoblasts. In this review, we summarized the different characteristics of the bone immune microenvironment of the jawbone compared with other bones, and the role of immune microenvironment regulation in common jawbone diseases. The development of corresponding therapeutic strategies for jawbone immune regulation targets may be helpful for the treatment of jawbone inflammatory diseases.

[Role of m 6A Reader YTHDC2 in Differentiation of Human Bone Marrow Mesenchymal Stem Cells].

OBJECTIVE: To study the regulatory effect of YTH domain-containing protein 2 (YTHDC2), a member of N 6-methyladenosine (m 6A) readers, on the osteogenic or adipogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). METHODS: YTHDC2 expression was knocked down by small interfering RNA (siRNA) in vitro. Osteogenic differentiation and adipogenic differentiation of hBMSCs were induced after YTHDC2 knockdown in order to study the changes in the differentiation phenotype of hBMSCs. Alkaline phosphatase staining (ALP staining) and alizarin red S staining were performed to examine osteogenic activity and calcium-nodular formation. Nile red staining was performed to examine lipid-droplet formation. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to assess the expression of osteogenesis and adipogenesis-related genes. RNA-sequencing was performed to identify the transcriptome changes after YTHDC2 knockdown and to explore the potential regulatory mechanism by which YTHDC2 regulated the diferentiation of hBMSCs. RESULTS: In this study, we found that siRNA-induced YTHDC2 knockdown resulted in increased ALP activity and calcium-nodular formation of hBMSCs during osteogenic differentiation, and significantly upregulated the expression of osteogenesis-related genes. In addition, the lipid-droplet formation capacity of hBMSCs was decreased during adipogenic differentiation. The expression of adipogenesis-related genes was significantly down-regulated. Gene-set enrichmen analysis of RNA-seq data showed that YTHDC2 was significantly correlated with ribosome function and mRNA-translation-related signaling pathways. CONCLUSION: The findings indicate that YTHDC2 knockdown can promote the osteogenic differentiation of hBMSCs and inhibit the adipogenic differentiation. YTHDC2 knockdown may cause changes in ribosome function.

Comparison of Artificial Neural Networks and Response Surface Methodology towards an Efficient Ultrasound-Assisted Extraction of Chlorogenic Acid from Lonicera japonica.

Chlorogenic acid (CGA), a bioactive compound commonly found in plants, has been demonstrated possessing nutraceutical potential in recent years. However, the more critical issue concerning how to improve production efficacy of CGA is still limited. It is a challenge to harvest a large amount of CGA without prolonging extraction time. In this study, the feasibility of using ultrasound for CGA extraction from Lonicera japonica was investigated. A central composite design (CCD) was employed to evaluate the effects of the operation parameters, including temperature, ethanol concentration, liquid to solid ratio, and ultrasound power on CGA yields. Meanwhile, the process of ultrasound-assisted extraction was optimized through modeling response surface methodology (RSM) and artificial neural network (ANN). The data indicated that CGA was efficiently extracted from the flower of Lonicera japonica by ultrasound assistance. The optimal conditions for the maximum extraction of CGA were as follows: The temperature at 33.56 °C, ethanol concentration at 65.88%, L/S ratio at 46:1 mL/g and ultrasound power at 150 W. ANN possessed greater optimization capacity than RSM for fitting experimental data and predicting the extraction process to obtain a maximum CGA yield. In conclusion, the process of ultrasound-assisted extraction can be well established by a methodological approach using either RSM or ANN, but it is worth mentioning that the ANN model used here showed the superiority over RSM for predicting and optimizing.

Statistical optimization of alkaline protease production from Penicillium citrinum YL-1 under solid-state fermentation.

Proteases from halotolerant and halophilic microorganisms were found in traditional Chinese fish sauce. In this study, 30 fungi were isolated from fermented fish sauce in five growth media based on their morphology. However, only one strain, YL-1, which was identified as Penicillium citrinum by internal transcribed spacer (ITS) sequence analysis, can produce alkaline protease. This study is the first to report that a protease-producing fungus strain was isolated and identified in traditional Chinese fish sauce. Furthermore, the culture conditions of alkaline protease production by P. citrinum YL-1 in solid-state fermentation were optimized by response surface methodology. First, three variables including peptone, initial pH, and moisture content were selected by Plackett-Burman design as the significant variables for alkaline protease production. The Box-Behnken design was then adopted to further investigate the interaction effects between the three variables on alkaline protease production and determine the optimal values of the variables. The maximal production (94.30 U/mL) of alkaline protease by P. citrinum YL-1 took place under the optimal conditions of peptone, initial pH, and moisture content (v/w) of 35.5 g/L, 7.73, and 136%, respectively.

Correlation between fluorescence in situ hybridization and testicular biopsy for the prediction of spermatogenesis in 37 patients with nonobstructive azoospermia.

OBJECTIVES: We applied interphase fluorescence in situ hybridization (FISH) to testis sections to examine the evidence of spermatogenesis in patients with nonobstructive azoospermia. This technique was evaluated and compared with conventional testicular histopathologic findings for the possibility of additional clinical applications. METHODS: Thirty-seven consecutive patients with nonobstructive azoospermia were carefully evaluated clinically. Testes were biopsied for both sperm extraction and histopathologic examination. FISH staining was performed with a CEP 18 SpectrumAqua/CEP X SpectrumGreen/CEP Y SpectrumOrange probe. RESULTS: Eight of 11 cases (sensitivity 73%) that were found to have spermatids on the histopathologic slides also were proven to produce haploid cells by FISH staining. On the other hand, 21 of the 26 cases (specificity 81%) for which no spermatids could be found on the histopathologic slides also had only diploid cells by FISH staining. On the basis of the good correlation between the FISH staining and conventional histopathologic findings, we could confirm the diagnosis of spermatogenesis using both methods. CONCLUSIONS: FISH staining of testicular sections allows more reliable prediction of spermatogenesis and provides benefits for a patient's decision regarding fertility counseling.