Probiotic characterization of Bacillus smithii: Research advances, concerns, and prospective trends.

Bacillus smithii is a thermophilic Bacillus that can be isolated from white wine, hot spring soil, high-temperature compost, and coffee grounds, with various biofunctions and wide applications. It is resistant to both gastric acid and high temperature, which makes it easier to perform probiotic effects than traditional commercial probiotics, so it can maintain good vitality during food processing and has great application prospects. This paper starts with the taxonomy and genetics and focuses on aspects, including genetic transformation, functional enzyme production, waste utilization, and application in the field of food science as a potential probiotic. According to available studies during the past 30 years, we considered that B. smithii is a novel class of microorganisms with a wide range of functional enzymes such as hydrolytic enzymes and hydrolases, as well as resistance to pathogenic bacteria. It is available in waste degradation, organic fertilizer production, the feed and chemical industries, the pharmaceutical sector, and food fortification. Moreover, B. smithii has great potentials for applications in the food industry, as it presents high resistance to the technological processes that guarantee its health benefits. It is also necessary to systematically evaluate the safety, flavor, and texture of B. smithii and explore its biological mechanism of action, which is of great value for further application in multiple fields, especially in food and medicine.

FoxO signaling pathway stimulation by Bacillus smithii XY1 contributes to alleviating copper-induced neurotoxicity.

Increasingly copper pollution in the environment exacerbates the risk of neurodegenerative diseases. It is necessary to look for effective targets and safe methods for protecting from copper-induced neurotoxicity. Here we firstly explored the impact of copper-exposure on expression profiles in zebrafish. Copper reduced embryo hatching, increased mortality and caused embryonic developmental abnormalities and behavioral dysfunction in juveniles. Transcriptomic analysis revealed that differential genes related to neuron were highly associated with oxidative stress especially enriched to FoxO pathway. Through further validation in Caenorhabditis elegans, copper resulted in nematode neurodegenerative movement disorders and neuronal damage, along with increased levels of reactive oxygen species (ROS) as well as decreased expressions of antioxidant-related enzymes and downstream genes which was also involved in FoxO signaling pathway. Bacillus smithii XY1, a novel strain with an excellent antioxidative activity, showed a great alleviative effect on copper-induced neurotoxicity that was related to FoxO stimulation, being a potential candidate for copper pollution management. Overall, these results suggested that FoxO pathway activation can regard as a strategy for mitigating neurotoxicity caused by copper and B. smithii XY1 with excellent tolerance and outstanding antioxidation specially targeted for FoxO has a promising application in controlling copper contamination.

Bacillus coagulans (Weizmannia coagulans) XY2 attenuates Cu-induced oxidative stress via DAF-16/FoxO and SKN-1/Nrf2 pathways and gut microbiota regulation.

Copper (Cu) pollution has become a serious environmental problem especially in recent decades. In this study, the mechanisms of Bacillus coagulans (Weizmannia coagulans) XY2 against Cu-induced oxidative stress were explored through a dual model. In mice, Cu disturbed microbial community structure, revealing an increased level of Enterorhabdus abundance and decreased levels of Intestinimonas, Faecalibaculu, Ruminococcaceae and Coriobacteriaceae_UCG-002 abundance. Meanwhile, B. coagulans (W. coagulans) XY2 intervention reversed this trend along with alleviated Cu-induced metabolic disturbances by increasing levels of hypotaurine and L-glutamate and declining levels of phosphatidylcholine and phosphatidylethanolamine. In Caenorhabditis elegans, nuclear translocation of DAF-16 and SKN-1 was inhibited by Cu, which in turn suppressed antioxidant-related enzymes activities. XY2 mitigated biotoxicity associated with oxidative damage caused by Cu exposure via regulating DAF-16/FoxO and SKN-1/Nrf2 pathways and intestinal flora to eliminate excess ROS. Our study provides a theoretical basis formulating future strategy of probiotics against heavy metal contamination.

Bacillus coagulans XY2 ameliorates copper-induced toxicity by bioadsorption, gut microbiota and lipid metabolism regulation.

Excessive copper pollutes the environment and endangers human health, attracting plenty of global attention. In this study, a novel strain named Bacillus coagulans XY2 was discovered to have a great copper tolerance and adsorption capacity. B. coagulans XY2 might maintain copper homeostasis through multisystem synergies of copper resistance, sulfur metabolism, Fe-S cluster assembly, and siderophore transport. In mice, by promoting the expression of SREBF-1 and SREBF-2 and their downstream genes, B. coagulans XY2 significantly inhibited the copper-induced decrease in weight growth rate, ameliorated dyslipidemia, restored total cholesterol and triglyceride contents both in serum and liver. Furthermore, B. coagulans XY2 recovered the diversity of gut microbiota and suppressed the copper-induced reduction in the ratio of Firmicutes to Bacteroidota. Serum metabolomics analysis showed that the alleviating effect of B. coagulans XY2 on copper toxicity was mainly related to lipid metabolism. For the first time, we demonstrated mechanisms of copper toxicity mitigation by B. coagulans XY2, which was related to self-adsorption, host copper excretion promotion, and lipid metabolism regulation. Moreover, working model of B. coagulans XY2 on copper homeostasis was predicted by whole-genome analysis. Our study provides a new solution for harmfulness caused by copper both in human health and the environment.

Chaihu Longgu Muli Decoction for post-stroke insomnia: A protocol for systematic review and meta-analysis.

BACKGROUND: Poststroke insomnia (PSI) is a frequent complication of stroke usually as a comorbidity of poststroke depression and mainly occurs within the first 6 months after stroke.[1] Addressing PSI to improve stroke prognosis is of great value. Herbal medicine like Chaihu Longgu Muli Decoction (CLMD), which is commonly considered to be a good treatment for depression and epilepsy, has the therapeutic potential on PSI; however, insufficient systematic reviews were conducted to testify its efficacy. Therefore, the objective of this paper is to provide reliable evidence of the efficacy and safety of CLMD on PSI and a foundation for further investigation. METHODS: The literature of clinical randomized controlled trials (RCTs) regarding CLMD for PSI published before June of 2021 will be retrieved in the databases, and 2 investigators will be asked to collect and crosscheck the data independently. For the including studies, the quality evaluation on methodology will be assessed in the light of the Cochrane Handbook for Systematic Review of Interventions V.5.1.0 as well as the quality of evidence will be evaluated by the Grading of Recommendations Assessment, Development, and Evaluation. Besides, the assessment of heterogeneity and reporting bias, the sensitivity analysis and the subgroup analysis will be conducted. Stata 15 will be applied to analyze the above data. RESULTS: The review will conduct a high-quality synthesis on present evidence of CLMD for PSI. CONCLUSION: The conclusion of the study will indicate whether CLMD is effective and safe for PSI.

Alleviative effects of a novel strain Bacillus coagulans XY2 on copper-induced toxicity in zebrafish larvae.

Copper (Cu) is a kind of micronutrient element that is essential for human metabolism. However, it is also considered as an environmental pollutant which is toxic to organisms at a high concentration level. Probiotics, regarded as beneficial microorganisms for promoting human health, have functions of antioxidant capacity, immune-enhancing properties, intestinal barrier protection and regulation. Several studies have reported that probiotics show positive effects on alleviating and intervening heavy metals toxicity. However, evidence for relieving copper-induced toxicity by probiotics is still limited. In this study, we firstly conducted a zebrafish larvae model to screen out microorganisms which are helpful for CuSO4 toxicity resistance and one novel strain named as Bacillus coagulans XY2 was discovered with the best protective activity. B. coagulans XY2 significantly reduced the mortality of zebrafish larvae exposed to 10 µmol/L CuSO4 for 96 hr, as well as alleviated the neutrophils infiltration in the larvae lateral line under a 2 hr exposure. B. coagulans XY2 exhibited a high in vitro antioxidant activity and against CuSO4-induced oxidative stress in zebrafish larvae by up-regulating sod1, gstp1 and cat gene transcriptional levels and relevant enzymatic activities. CuSO4 stimulated the inflammation process resulting in obvious increases of gene il-1ß and il-10 transcription, which were suppressed by B. coagulans XY2 intervention. Overall, our results underline the bio-function of B. coagulans XY2 on protecting zebrafish larvae from copper toxicity, suggesting the potential application values of probiotics in copper toxicity alleviation on human and the environment.

Effect of different acupuncture and moxibustion methods on functional dyspepsia caused by sequelae of COVID-19: A protocol for systematic review and meta-analysis.

BACKGROUND: Functional dyspepsia (FD) is a group of diseases that cannot be explained after routine clinical examination, and is characterized by postprandial fullness, early satiety, and upper abdominal pain or burning. According to the statistics, FD continues to become one of the high-risk sequelae of coronavirus disease 2019 (COVID-19), affecting patients' quality of life, increasing psychological burden and increasing economic costs. However, its optimal treatment is still an urgent problem. A large number of studies have shown that acupuncture and moxibustion is effective and safe in the treatment of FD caused by sequelae of COVID-19, which is of research value. Therefore, based on the current literatures, the effectiveness and safety of different acupuncture and moxibustion methods were systematically evaluated to provide possible alternative therapy on FD. METHODS: Studies search for eligible randomized controlled trials that use different acupuncture and moxibustion methods as the sole treatment on FD and their data extraction will be done by 2 researchers. In case of disagreement, a third researcher will be introduced for arbitration. Mean difference or relative risk with fixed or random effect model in terms of 95% confidence interval will be adopted for the data synthesis. To evaluate the risk of bias, the Cochrane risk of bias assessment tool will be utilized. The sensitivity or subgroup analysis will also be conducted when meeting high heterogeneity (I2 > 50%). RESULTS: This meta-analysis will provide an authentic synthesis of different acupuncture and moxibustion methods on FD caused by sequelae of COVID-19. CONCLUSION: This meta-analysis will evaluate the effect of acupuncture and moxibustion on FD caused by sequelae of COVID-19, providing evidence as to the treatment in these patients.

A Rapid Screening Method of Candidate Probiotics for Inflammatory Bowel Diseases and the Anti-inflammatory Effect of the Selected Strain Bacillus smithii XY1.

Inflammatory bowel disease (IBD) is a chronic intestinal disease associated with the inflammatory gastrointestinal tract and microbiome dysbiosis. Probiotics are a promising intervention, and several probiotics have been reported to positively affect IBD remission and prevention, particularly on ulcerative colitis (UC). However, there is still a limitation in the knowledge of effectiveness and safety of probiotics therapies for IBD. Exploring more potential probiotics helps to find extensive evidence for probiotic intervention. This study established a rapid method for probiotics candidate screening and finally screened out one strain with the best protective effect. Forty strains isolated from four different sources were used for this screening. Hemolysis tests and acute toxic test evaluated strain safety. Zebrafish were first treated with dextran sodium sulfate (DSS) for colitis induction, and every bacteria were individually added to the fish water subsequently. Results showed eight strains could lower the larvae mortality within 3 days under a 0.6% DSS concentration, including Lacticaseibacillus rhamnosus GG, L. rhamnosus NBRC3425, Bacillus smithii DSM4216, B. smithii XY1, Bacillus coagulans NBRC12583, Bacillus coagulans XY2, Lactobacillus parafarraginis XYRR2, and Bacillus licheniformis XYT3. Among eight, B. smithii XY1 was the only strain having the equal ability to alleviate neutrophil infiltration in the larvae intestine with that ability of prednisolone under a 0.5% DSS concentration. Bacillus smithii XY1 restored intestinal epithelial cell integrity after DSS damage, as well as regulated the gene expression inflammation-related factors, indicating its bio-function of inflammatory response alleviation.

Efficient biodegradation of organic matter using a thermophilic bacterium and development of a cost-effective culture medium for industrial use.

Microorganisms with efficient organic matter degradation ability are essential for organic waste treatment. In this study, a thermophilic bacterium, Bacillus thermoliquefaciens, was identified to have excellent cellulase, amylase, and protease activity, and provided efficient degradation of food waste. This is the first report on the organic matter degradation potential of B. thermoliquefaciens. Using a "one-variable-at-a-time" approach and response surface methodology, the optimal culture conditions for B. thermoliquefaciens were determined to be a 5% inoculation level, 50 °C culture temperature, 25 mL filling volumes in 250 mL flasks, and 180 rpm shaking for 24 h. The optimized medium was formulated as 1 g Na2HPO4, 1 g KH2PO4, 0.05 g MgSO4, 3 g NaCl, 0.05 g CaCl2, 11.44 g wheat bran powder, 4.92 g soybean meal, and 1 L distilled water at pH 7.12. The maximum biomass attained was 1.57 ± 0.153 × 109 CFU/mL. The cost of this medium was 4.18 times less than that before optimization. This promising result lays a foundation for future industrial application of this bacterium to the degradation of organic waste.

A Mechanism Study of NMDAR1 in A Rat Alzheimer Disease (AD) Model

ABSTRACT This study aimed to investigate the expression and mechanism of N- methyl -D- aspartate receptor 1 (NMDAR1) in the pathogenesis of Alzheimer disease (AD). Eighty adult Wistar rats were randomly divided into 4 groups (n=20 each) to receive an injection of 0, 5, 7 and 10 μl of 1 μg/μl amyloid-β 42 (Aβ1-42) in the hippocampus. Twenty rats in normal control group were injected with equal volume of saline. After 10 days, the hippocampus was isolated from 5 randomly selected rats in each group. The NMDAR1 protein and mRNA expression was determined by immunohistochemical staining and qRT-PCR. The aquaporin-1 (AQP-1) mRNA expression was also measured by qRT-PCR. We found that both NMDAR1 and AQP-1 expression in Aβ1-42 groups was increased in a dose-dependent manner. NMDAR1 and AQP-1 expression in 7 and 10 μl Aβ1-42 groups was significantly higher compared with 0 μl Aβ1-42 group (P <0.01). Further, the 10 μl Aβ1-42 group was randomly divided into 3 subgroups: AD-NMDA, AD-MK-801, and AD-Ctrl subgroup, which was given an intraperitoneal injection of NMDAR agonist NMDA, NMDAR antagonist MK-801 and saline, respectively. The relative APQ-1 expression in each subgroup was determined by qRT-PCR and Western blot analysis after 24 h. The AQP-1 expression was significantly decreased in AD-MK-801 group (P < 0.05), but was markedly increased in AD-NMDA group when compared with AD-Ctrl group (P <0.01). Our study suggested that expression abnormity of NMDAR1 is involved in the pathogenesis of AD. NMDAR1 might regulate the pathogenic process through stimulating the expression of AQP-1.

[Effects of Marsdenia tenacissima extract on proliferation and apoptosis of hematologic neoplasm cell line cells].

OBJECTIVE: To investigate the effects of the extract from Marsdensia tenacissima on proliferation and apoptosis of human hematologic neoplasm cell line cells. METHODS: Raji, NB4 and K562 cells were treated in vitro with different concentrations of the extract from Marsdensia tenacissima, including different ethanol elution components and C21 steroidal saponin monomer compounds, for different periods. Tumor cell proliferation was measured by MTT colorimetric assay and its apoptosis was determined by the flow cytometry (FCM). RESULTS: Firstly, with higher concentrations, 100 microg/mL and 200 microg/mL, 70% ethanol eluate from Marsdensia tenacissima inhibited the proliferation of Raji, NB4 and K562 cells significantly, in a dose and time dependent manner, compared with 30% and 50% ethanol elution components from Marsdensia tenacissima (P < 0.05). Secondly, four C21 steroidal saponin monomer compounds, tenacissosides B,C,I and marsdenoside K, also inhibited the proliferation of Raji, NB4 and K562 cells in vitro significantly, in a dose and time dependent manner, compared with that of control group (P < 0.05). Among them, tenacissoside C showed the strongest inhibition effects on proliferation of these cells under all experimental conditions compared with the other three C21 steroidal saponin monomer compounds (P < 0.05). Furthermor, the IC50 of tenacissosides C on proliferation of Raji, NB4 and K562 cells were 64.1 micromol/L, 70.4 micromol/L and 105.8 micromol/L, respectively. Finally, after Raji, NB4 and K562 cells were treated with 98.4 micromol/L tenacissoside C for 24 h and 48 h, the early apoptosis rates and late apoptosis rates of these tumor cells increased markedly, compared with the control group (P < 0.05). CONCLUSION: The extract from Marsdensia tenacissima, including different ethanol elution components and C21 steroidal saponin monomer compounds, may inhibit the proliferation of some human hematologic neoplasm cell line cells and induce these tumor cells apoptosis in vitro, especially tenacissoside C, one of the C21 steroidal saponin monomer compounds, showed the strongest effects on proliferation of these tumor cells when compared with other ones, with the strongest inhibition activities on human Burkitt's lymphoma cell line Raji cells.