Live Bacillus Subtilis Research Articles

Molecular Characterization of Ancient Prosaposin-like Proteins from the Protist Dictyostelium discoideum.

To combat the permanent exposure to potential pathogens every organism relies on an immune system. Important factors in innate immunity are antimicrobial peptides (AMPs) that are structurally highly diverse. Some AMPs are known to belong to the saposin-like proteins (SAPLIPs), a group of polypeptides with a broad functional spectrum. The model organism Dictyostelium discoideum possesses a remarkably large arsenal of potential SAPLIPs, which are termed amoebapore-like peptides (Apls), but the knowledge about these proteins is very limited. Here, we report about the biochemical characterization of AplE1, AplE2, AplK1, and AplK2, which are derived from the two precursor proteins AplE and AplK, thereby resembling prosaposins of vertebrates. We produced these Apls as recombinant polypeptides in Escherichia coli using a self-splicing intein to remove an affinity tag used for purification. All recombinant Apls exhibited pore-forming activity in a pH-dependent manner, as evidenced by liposome depolarization, showing higher activities the more acidic the setting was. Lipid preference was detected for negatively charged phospholipids and in particular for cardiolipin. Antimicrobial activity against various bacteria was found to be inferior in classical microdilution assays. However, all of the Apls studied permeabilized the cytoplasmic membrane of live Bacillus subtilis. Collectively, we assume that the selected Apls interact by their cationic charge with negatively charged bacterial membranes in acidic environments such as phagolysosomes and eventually lyse the target cells by pore formation.

Immunomodulatory effects of live and UV-killed Bacillus subtilis natto on inflammatory response in human colorectal adenocarcinoma cell line in vitro.

Colorectal cancer (CRC) is a heterogeneous disease of the colon or rectum arising from adenoma precursors and serrated polyps. Recently, probiotics have been proposed as an effective and potential therapeutic approach for CRC prevention and treatment. Probiotics have been shown to alleviate inflammation by restoring the integrity of the mucosal barrier and impeding cancer progression. In this study, we aimed to investigate the immunomodulatory effects of live and UV-killed Bacillus subtilis natto on the inflammatory response in CRC. Caco-2 cells were exposed to various concentrations of live and UV- killed B. subtilis natto, and cell viability was assessed using MTT assay. Gene expression analysis of IL-10, TGF-β, TLR2 and TLR4 was performed using RT-qPCR. Our findings showed that both live and UV-killed B. subtilis natto caused significant reduction in inflammatory response by decreasing the gene expression of TLR2 and TLR4, and enhancing the gene expression of IL-10 and TGF-β in Caco-2 cells as compared to control group. The results of this study suggest that live and UV-killed B. subtilis natto may hold potential as a therapeutic supplement for modulating inflammation in CRC.

Self-organization of mortal filaments and its role in bacterial division ring formation

Filaments in the cell commonly treadmill. Driven by energy consumption, they grow on one end while shrinking on the other, causing filaments to appear motile even though individual proteins remain static. This process is characteristic of cytoskeletal filaments and leads to collective filament self-organization. Here we show that treadmilling drives filament nematic ordering by dissolving misaligned filaments. Taking the bacterial FtsZ protein involved in cell division as an example, we show that this mechanism aligns FtsZ filaments in vitro and drives the organization of the division ring in living Bacillus subtilis cells. We find that ordering via local dissolution also allows the system to quickly respond to chemical and geometrical biases in the cell, enabling us to quantitatively explain the ring formation dynamics in vivo. Beyond FtsZ and other cytoskeletal filaments, our study identifies a mechanism for self-organization via constant birth and death of energy-consuming filaments.

Direct observation of a crescent-shape chromosome in expanded Bacillus subtilis cells

Bacterial chromosomes are folded into tightly regulated three-dimensional structures to ensure proper transcription, replication, and segregation of the genetic information. Direct visualization of chromosomal shape within bacterial cells is hampered by cell-wall confinement and the optical diffraction limit. Here, we combine cell-shape manipulation strategies, high-resolution fluorescence microscopy techniques, and genetic engineering to visualize the shape of unconfined bacterial chromosome in real-time in live Bacillus subtilis cells that are expanded in volume. We show that the chromosomes predominantly exhibit crescent shapes with a non-uniform DNA density that is increased near the origin of replication (oriC). Additionally, we localized ParB and BsSMC proteins – the key drivers of chromosomal organization – along the contour of the crescent chromosome, showing the highest density near oriC. Opening of the BsSMC ring complex disrupted the crescent chromosome shape and instead yielded a torus shape. These findings help to understand the threedimensional organization of the chromosome and the main protein complexes that underlie its structure.

Antioxidative single atomic nanocatalysts facilitate orally administered probiotic inulin gels for acute colitis amelioration

Acute colitis is characterized by the microenvironment with a high level of inflammation, oxidative stress as well as a disordered microbiome. Sustained delivery of the therapeutic medicine into the colitis foci is limited due to the harsh condition of the gastric environment and fast excretion kinetics in the gut. Herein, we have designed an orally administrable probiotic inulin gel (igel) containing living Bacillus subtilis (B. subt) engineered with outperformed antioxidative single manganese atomic nanocatalyst (SAM NCs) for effective, sustained, and biocompatible therapeutics against acute colitis. Based on the multi-catalytic antioxidative activities (superoxide dismutase, glutathione peroxidase, and catalase) of the synthetic SAM NCs, these nanocatalysts are capable of eliminating the oxidative stresses both in vitro and in vivo. With further engineering onto the living Bacillus subtilis and subsequent in situ gelation inside the inulin, the as-formed probiotic igel(SAM NCs@B. subt) could effectively target and colonize inside the colon region, triggering the in vivo anti-inflammatory, anti-oxidative and gut microbiome modulation performance with high biocompatibility. The present study provides a promising probiotic gelation therapeutic strategy against acute colitis, shedding light on the exploration and clinical transformation of gastrointestinal disease.

Biophotoelectrochemical process co-driven by dead microalgae and live bacteria.

Anaerobic reduction processes in natural waters can be promoted by dead microalgae that have been attributed to nutrient substances provided by the decomposition of dead microalgae for other microorganisms. However, previous reports have not considered that dead microalgae may also serve as photosensitizers to drive microbial reduction processes. Here we demonstrate a photoelectric synergistic linkage between dead microalgae and bacteria capable of extracellular electron transfer (EET). Illumination of dead Raphidocelis subcapitata resulted in two-fold increase in the rate of anaerobic bioreduction by pure Geobacter sulfurreducens, suggesting that photoelectrons generated from the illuminated dead microalgae were transferred to the EET-capable microorganisms. Similar phenomena were observed in NO3- reduction driven by irradiated dead Chlorella vulgaris and living Shewanella oneidensis, and Cr(VI) reduction driven by irradiated dead Raphidocelis subcapitata and living Bacillus subtilis. Enhancement of bioreduction was also seen when the killed microalgae were illuminated in mixed-culture lake water, suggesting that EET-capable bacteria were naturally present and this phenomenon is common in post-bloom systems. The intracellular ferredoxin-NADP+-reductase is inactivated in the dead microalgae, allowing the production and extracellular transfer of photoelectrons. The use of mutant strains confirmed that the electron transport pathway requires multiheme cytochromes. Taken together, these results suggest a heretofore overlooked biophotoelectrochemical process jointly mediated by illumination of dead microalgae and live EET-capable bacteria in natural ecosystems, which may add an important component in the energetics of bioreduction phenomena particularly in microalgae-enriched environments.

Quantification of the antagonistic and synergistic effects of Pb2+, Cu2+, and Zn2+ bioaccumulation by living Bacillus subtilis biomass using XGBoost and SHAP

Bioaccumulation and adsorption are efficient methods for removing heavy metal ions (HMIs) from aqueous environments. However, methods to quantifiably characterize the removal selectivity for co-existing HMIs are limited. In this study, we applied Shapley additive explanations (SHAP) following extreme gradient boosting (XGBoost) modeling, to generate SHAP values. We used these values to create an affinity interference index (AII) that quantitatively represented the interference between metal ions in a multi-metal bioaccumulation system. The selectivity for simultaneous bioaccumulation of Pb2+, Cu2+, and Zn2+ by living Bacillus subtilis biomass was then characterized as a proof of concept. The AII indicated that the bioaccumulation of Zn2+ was more strongly inhibited by Pb2+/Cu2+ (AII = 1) than that of Pb2+/Cu2+ by Zn2+. Moreover, the presence of Zn2+ promoted the bioaccumulation of Pb2+ (AII = 0.39), which was confirmed in further experiments where the bioaccumulation of Pb2+ (300 μM) was increased by 38% with Zn2+ (300 μM). This study demonstrated that the combination of XGBoost and SHAP is effective in the quantifiable characterization of the antagonistic and synergistic effects in a multi-metal simultaneous bioaccumulation system. This method could also be generalized to similar tasks for analyzing the selectivity effects in a multi-component system.

Oral administration of live combined Bacillus subtilis and Enterococcus faecium alleviates colonic oxidative stress and inflammation in osteoarthritic rats by improving fecal microbiome metabolism and enhancing the colonic barrier.

Osteoarthritis (OA) causes intestinal damage. The protective effect of probiotics on the intestine is indeed effective; however, the mechanism of protection against intestinal damage in OA is not clear. In this study, we used meniscal/ligamentous injury (MLI) to mimic OA in rats and explored the colonic protective effects of Bacillus subtilis and Enterococcus faecium on OA. Our study showed that treatment with B. subtilis and E. faecium attenuated colonic injury and reduced inflammatory and oxidative stress factors in the serum of osteoarthritic rats. α- and ß diversity of the fecal flora were not different among groups; no significant differences were observed in the abundances of taxa at the phylum and genus levels. We observed the presence of the depression-related genera Alistipes and Paraprevotella. Analysis of fecal untargeted metabolism revealed that histamine level was significantly reduced in the colon of OA rats, affecting intestinal function. Compared to that in the control group, the enriched metabolic pathways in the OA group were primarily for energy metabolisms, such as pantothenate and CoA biosynthesis, and beta-alanine metabolism. The treatment group had enriched linoleic acid metabolism, fatty acid biosynthesis, and primary bile acid biosynthesis, which were different from those in the control group. The differences in the metabolic pathways between the treatment and OA groups were more evident, primarily in symptom-related metabolic pathways such as Huntington's disease, spinocerebellar ataxia, energy-related central carbon metabolism in cancer, pantothenate and CoA biosynthesis metabolic pathways, as well as some neurotransmission and amino acid transport, and uptake- and synthesis-related metabolic pathways. On further investigation, we found that B. subtilis and E. faecium treatment enhanced the colonic barrier of OA rats, with elevated expressions of tight junction proteins occludin and Zonula occludens 1 and MUC2 mRNA. Intestinal permeability was reduced, and serum LPS levels were downregulated in the treatment group. B. subtilis and E. faecium also regulated the oxidative stress pathway Keap1/Nrf2, promoted the expression of the downstream protective proteins HO-1 and Gpx4, and reduced intestinal apoptosis. Hence, B. subtilis and E. faecium alleviate colonic oxidative stress and inflammation in OA rats by improving fecal metabolism and enhancing the colonic barrier.

Effects of Live Combined Bacillus subtilis and Enterococcus faecium on Gut Microbiota Composition in C57BL/6 Mice and in Humans.

Probiotics, prebiotics, and synbiotics can alleviate metabolic syndrome by altering the composition of the gut microbiota. Live combined Enterococcus faecium and Bacillus subtilis has been indicated to promote growth and reduce inflammation in animal models. However, the modulatory effects of live combined B. subtilis R-179 and E. faecium R-026 (LCBE) on human microbiota remain unclear. The current study examined the growth of these two strains in the presence of various oligosaccharides and assessed the effects of this probiotic mixture on human and murine gut microbiota in vitro and in vivo. Oligosaccharides improved the growth of E. faecium R-026 and B. subtilis R-179 as well as increased their production of short-chain fatty acids. E. faecium R-026 or B. subtilis R-179 co-incubated with Bifidobacterium and Clostridium significantly increased the number of the anaerobic bacteria Bifidobacterium longum and Clostridium butyricum by in vitro fermentation. Moreover, LCBE significantly reduced plasma cholesterol levels in mouse models of hyperlipidemia. LCBE combined with galacto-oligosaccharides led to a significant decrease in the Firmicutes/Bacteroidetes ratio and a significant increase in the relative abundance of Akkermansia and Bifidobacteria after treating mice with LCBE (0.23 g/day) for eight weeks. Furthermore, in vitro fermentation also showed that both the single strains and the two-strain mixture modulated human gut microbiota, resulting in increased Lactobacillus and Bifidobacteria, and decreased Escherichia-Shigella. Overall, these results suggest that LCBE can improve host health by reducing the level of cholesterol in mouse models by modifying the composition of the gut microbiota.

Live Combined B. subtilis and E. faecium Alleviate Liver Inflammation, Improve Intestinal Barrier Function, and Modulate Gut Microbiota in Mice with Non-Alcoholic Fatty Liver Disease.

BackgroundNon-alcoholic fatty liver disease (NAFLD) is a chronic, progressive liver disease with an increasing incidence rate. This study investigated the protective effects of live combined Bacillus subtilis and Enterococcus faecium (LCBE) on NAFLD, and its possible mechanisms.Material/MethodsFive-week-old C57BL/6 mice were randomly divided into 3 groups: chow, HFD, and HFD+LCBE groups. The levels of serum biochemical markers, glucose tolerance, insulin, the inflammatory cytokines IL-1β, IL-6, and TNF-α, LPS, and histological staining were measured using commercial kits. qPCR was used to examine the mRNA expression levels of inflammatory cytokines in the liver. Western blotting was used to determine the protein levels of TLR4, NF-κB p65, PPAR-α, and CPT-1 in the liver, and occludin and Claudin1 in the intestine. The intestinal flora of the mice was analyzed by high-throughput sequencing of the V3–V4 region of 16S rDNA.ResultsLCBE significantly lowered the body weight, liver/body weight ratio, and serum glucose level, and increased the serum insulin level in NAFLD mice. In addition, LCBE treatment improved the liver function and lipid profile, decreased the levels of LPS and inflammatory cytokines, and downregulated the expression of TLR4 and NF-κB p65. Moreover, LCBE enhanced the intestinal barrier function by increasing the expression of occludin and Claudin1. Furthermore, LCBE modulated the composition of the gut microbiota by reducing the Firmicutes to Bacteroidetes ratio, and the proportion of inflammation-related and LPS-producing bacteria, thus re-arranging the structure of the gut microbiota.ConclusionsLCBE protects against NAFLD by alleviating inflammation, restoring the intestinal barrier, and modulating gut microbiota composition.

Bacillus subtilis in PVA Microparticles for Treating Open Wounds.

Open wound dressings should provide a moist environment, protect the wound from bacterial contamination, and shield it from further damage. These requirements, however, are hard to accomplish since such wounds are colonized by pathogenic bacteria, including resistant species such as methicillin-resistant Staphylococcus aureus (MRSA). A new approach for treating open wounds that is based on sticky and dissolvable polyvinyl alcohol (PVA) microparticles containing live Bacillus subtilis (B. subtilis) is described. Microparticles, fabricated by the spray-drying technique, were administered directly to an open wound while B. subtilis continuously produced and secreted antimicrobial molecules. B. subtilis in PVA microparticles demonstrated remarkable antibacterial activity against MRSA and S. aureus. In in vivo experiments, both B. subtilis and empty PVA microparticles were effective in decreasing healing time; however, B. subtilis microparticles were more effective during the first week. There was no evidence of skin irritation, infection, or other adverse effects during the 15 day postoperative observation period. This concept of combining live secreting bacteria within a supportive delivery system shows great promise as a therapeutic agent for open wounds and other infectious skin disorders.

Effect of Combined Live Probiotics Alleviating the Gastrointestinal Symptoms of Functional Bowel Disorders.

Objective Changes of the gut microbiota are related to the pathogenesis of functional bowel disorders (FBDs), and probiotic supplementation may be an effective treatment option. Therefore, we aimed to investigate the effect of combined live probiotics on the gastrointestinal symptoms of FBDs via altering the gut microbiota. Methods Patients with the gastrointestinal symptoms of FBDs attending the Outpatient Department, from July to November 2019, were recruited. After the bowel preparation with polyethylene glycol electrolyte powder and colonoscopy, patients with normal result of colonoscopy were randomly divided into the probiotics group and control group. Patients in the probiotics group were prescribed with combined live Bacillus subtilis and Enterococcus faecium enteric-coated capsules for 4 weeks. Small intestinal bacteria overgrowth (SIBO) was measured by lactulose hydrogen breath test, and the microbial DNA was extracted from the fecal samples and the bacteria were classified by 16S rDNA gene amplicon sequencing. Results Twenty-five patients of each group were recruited, and there was no significant difference between the probiotics and control groups on baseline gastrointestinal symptom rating scale (GSRS), positive rate of SIBO, and relative abundances of the gut microbiota at the phylum level. After 4 weeks of treatment, the values of the probiotics and control groups were as follows: GSRS 1.4 ± 1.4 and 3.6 ± 1.6 and positive rate of SIBO 28.0% and 56.0%, respectively. The median relative abundances of the gut microbiota were 1.01% and 5.03% Actinobacteria and 43.80% and 35.17% Bacteroidetes at the phylum level; 0.76% and 3.29% Bifidobacterium, 0.13% and 0.89% Cillinsella, 0.03% and 0.01% Enterococcus, 0.18% and 0.36% Lachnospiraceae, 0.10% and 0.16% Ruminococcus torques group, 1.31% and 2.44% Blautia, and 0.83% and 2.02% Fusicatenibacter at the genus level (P < 0.05), respectively. Conclusion Combined live probiotic supplementation after the bowel preparation can alter the gut microbiota, decontaminate SIBO, and alleviate the gastrointestinal symptoms of FBDs. This trial is registered with ChiCTR1900026472.

A retrospective study of probiotics for the treatment of children with antibiotic-associated diarrhea.

This retrospective study aimed to explore the benefits and safety of probiotics (live combined Bacillus subtilis and Enterococcus faecium granules with multivitamines) for the treatment of children with antibiotic-associated diarrhea (AAD).A total of 72 children with AAD were analyzed in this study. Of these, 36 children received routine treatment plus probiotics, and were assigned to a treatment group. The other 36 children underwent routine treatment alone, and were assigned to a control group. Patients in both groups were treated for a total of 7 days. The efficacy and safety were evaluated by duration of diarrhea (days), number of dressings needed daily, abdominal pain intensity, stool consistency (as assessed by Bristol Stool Scale (BSS)), and any adverse events.After treatment, probiotics showed encouraging benefits in decreasing duration of diarrhea (days) (P < .01), number of dressings needed every day (P < .01), abdominal pain intensity (P < .01), and stool consistency (BSS (3–5), P < .01; BSS (6–7), P < .01). In addition, no adverse events were documented in this study.The findings of this study demonstrated that probiotics may provide promising benefit for children with AAD. Further studies are still needed to warrant theses findings.

Mechanistic insight into the effect of BT-benzo-29 on the Z-ring in Bacillus subtilis.

The assembly and disassembly of FtsZ play an essential role in bacterial cell division. Using single-cell imaging, we report that short exposure to BT-benzo-29 inhibits Z-ring formation in live Bacillus subtilis cells. Fluorescence recovery after photobleaching of the Z-ring in live bacteria demonstrated that BT-benzo-29 strongly suppressed the assembly dynamics of FtsZ in the Z-ring. Furthermore, B. subtilis cells expressing V275A-FtsZ resisted the antibacterial activity of BT-benzo-29 providing evidence that BT-benzo-29 inhibits bacterial proliferation by targeting FtsZ. In addition, a brief (8 min) exposure of BT-benzo-29 destroyed the Z-ring without perturbing the localization of a late cell division protein, DivIVA, the nucleoid segregation, and membrane permeability. BT-benzo-29, when used in combination with vancomycin and polymyxin B (PMB), produced a much stronger inhibitory effect on Bacillus subtilis and Escherichia coli cells, respectively. The combination index of BT-benzo-29 with vancomycin and PMB was determined to be <1, suggesting that BT-benzo-29 exhibits synergistic inhibitory effects on bacterial proliferation when used along with these antibiotics.

Methods for Studying Membrane-Associated Bacterial Cytoskeleton Proteins In Vivo by TIRF Microscopy.

MreB proteins are actin homologs present in nonspherical bacteria. They assemble into membrane-associated discrete filamentous structures that exhibit different dynamic behaviors along the bacterial sidewalls. Total internal reflection fluorescence (TIRF) microscopy, a sensitive method for studying molecular events at cell surfaces with high contrast and temporal resolution, is a method of choice to characterize the localization and dynamics of cortical MreB assemblies in vivo. This chapter describes the methods for visualizing fluorescently tagged MreB proteins in live Bacillus subtilis cells. We detail how to (1) grow B. subtilis strains for reproducible TIRF observations, (2) immobilize cells on agarose pads and (3) in CellASIC® microfluidic plates, and (4) acquire TIRF images and time lapses.

Protective Effects of Live Combined B. subtilis and E. faecium in Polymicrobial Sepsis Through Modulating Activation and Transformation of Macrophages and Mast Cells.

Aims: Clinical studies showed that the use of probiotics during critical illness reduced nosocomial infection and improved clinical outcome. However, the functional mechanisms of probiotics is remains unclear. Therefore the aim of current study is to explore the protective effects and understand the underlying mechanisms for the beneficial effects of live combined Bacillus subtilis and Enterococcus faecium (LCBE) in cecal ligation puncture (CLP)-induced sepsis.Methods and Results: Seven-week-old C57BL/6J mice were divided into three groups: sham group (6 mice), CLP-control group (20 mice, pretreatment with saline for 7 days before CLP surgery) and CLP-probiotics group (14 mice, pretreatment with LCBE enteric-coated capsules for 7 days before CLP surgery). In survival experiment, mice were monitored for 7 days after CLP. After the procedure, mice were sacrificed, and, serum, and peritoneal lavage fluid were collected and intestinal ileal samples were harvested.Results: Our results showed that the mortality was significantly reduced in mice CLP-probiotics group vs. CLP-control group (P < 0.05). Also, treatment CLP-probiotics group decreased the injury scores CLP-probiotics group when compared to CLP-control group. Additionally, levels of pro-inflammatory cytokines IL-6 and TNF-α levels in the serum and intestinal ileal tissues of CLP-probiotics group were reduced when compared to CLP-control group (P < 0.05). However, no significant differences in anti-inflammatory levels of IL-10 and TGF-β1 were observed between CLP-control and CLP-probiotic groups. Furthermore, our experiments showed that that probiotic treatment suppressed the macrophage activation and transformation from M-type to M1-type, inhibited the mast cells (MCs) degranulation, and activation of AKT (kinase B) pathway.Conclusion: In conclusion, our data shows that probiotics have a protective role in CLP septic mice through reducing intestinal inflammation, altering macrophage polarization and MCs degranulation, and regulating AKT signaling.Significance and Impact of Study: This study demonstrated the protective effects and mechanisms involved in the protective role of live combined Bacillus subtilis and Enterococcus faecium (LCBE) in CLP-induced septic mice model.

Dynamic Exchange of Two Essential DNA Polymerases during Replication and after Fork Arrest

The replisome is a multiprotein machine responsible for the faithful replication of chromosomal and plasmid DNA. Using single-molecule super-resolution imaging, we characterized the dynamics of three replisomal proteins in live Bacillus subtilis cells: the two replicative DNA polymerases, PolC and DnaE, and a processivity clamp loader subunit, DnaX. We quantified the protein mobility and dwell times during normal replication and following replication fork stress using damage-independent and damage-dependent conditions. With these results, we report the dynamic and cooperative process of DNA replication based on changes in the measured diffusion coefficients and dwell times. These experiments show that the replication proteins are all highly dynamic and that the exchange rate depends on whether DNA synthesis is active or arrested. Our results also suggest coupling between PolC and DnaX in the DNA replication process and indicate that DnaX provides an important role in synthesis during repair. Furthermore, our results suggest that DnaE provides a limited contribution to chromosomal replication and repair in vivo.

Living Bacteria in Thermoresponsive Gel for Treating Fungal Infections

AbstractThe leading living bacteria formulations currently available are from a limited list of genera and are generally limited to gastrointestinal tract syndromes. A formulation composed of living Bacillus subtilis incorporated in a thermoresponsive hydrogel that hardens after administration on the skin and continuously produces antifungal agents is described. The ability of the formula to support bacteria growth and its mechanical properties and penetrability through the skin are fine‐tuned by varying the ratio between polymer concentrations and bacterial media. The formula penetrates via the stratum corneum and accumulates in the epidermis without penetrating the inner, dermis layer. In vivo results mirror the results seen in vitro: bacillus formulations completely inhibit candida growth, demonstrating clinical effects comparable to those achieved by ketoconazole. LC‐MS/MS analysis of the bacterial formulation confirms the presence of surfactin, the most powerful biosurfactant that possesses a broad antifungal activity. This platform may enable rational design of novel formulations composed of secreting bacteria inside a responsive, smart, hydrogel—which is the prerequisite for producing a successful drug delivery system.

Efficacy of probiotics on the treatment of non-alcoholic fatty liver disease

Objective: To study the clinical effect of probiotics in the treatment of non-alcoholic fatty liver disease (NAFLD). Methods: A total of 200 patients with NAFLD were randomly divided into 4 groups: control group (routine treatment group) and combined treatment group A, B and C. Each group had equal patients. The control group received orally polyene phosphatidylcholine capsules; whereas combined group A, B and C were given orally the live "combined Bifidobacterium Lactobacillus and Enterococcus powder" , "two live combined Bacillus subtilis and Enterococcus" , and the both probiotics respectively. The duration of treatment was 1 month. Laboratory parameters were evaluated before treatment and thirtieth day after treatment, including cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol(LDL-C), alanine aminotransferase(ALT), aspartate aminotransferase (AST), fasting blood glucose (FPG), serum high molecular weight adiponectin (HMW-APN) and serum TNFα. Meanwhile the faece sample was collected for routine test and bacterial culture. Liver ultrasound scan was done in all patients. Results: In terms of blood lipids and blood glucose, each group improved after treatment with significant differences (P<0.05) except for HDL-C. As for liver function, serum ALT and AST decreased after treatment in each group; especially in combined group C which were lower than those of control group [(33.7±7.6) U/L vs. (45.0±8.5) U/L; (22.0±1.6) U/L vs. (29.4±3.7) U/L; P<0.05]. TNFα levels decreased after treatment in each group, in addition the values in combined group C was significantly lower than that of control group[(0.51±0.27) µg/L vs. (0.82±0.28) µg/L, P<0.05]. Serum HMW-APN increased after treatment in each group, and the HMW-APN in combined C group was significantly higher than that of control group[(9.28±3.72) µg/L vs. (7.87±3.96)µg/L, P<0.05]. (5) After treatment, all groups showed improvement of fatty liver by ultrasound, but the difference between groups was not statistically significant. (6) Compared with before treatment, fecal flora in combined groups was all reduced (P<0.01), but it was comparable before and after treatment in control group. Conclusions: Probiotics improve intestinal microecological system in NAFLD patients via inhibiting TNFα and enhancing adiponectin, possibly resulting in regulating blood glucose, lipid metabolism, and protecting liver injury from NAFLD.

The clinical effect of live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice

Objective To explore the clinical effect of live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice. Methods 90 patients with neonatal jaundice were randomly divided into control group and observation group.The control group received the blue light treatment, the observation group was given blue light irradiation combined with live Bacillus Subtilis and Enterococcus Faecium granules treatment, the patients were treated for 7 days.The clinical efficacy, serum bilirubin and main indicators of complex constant time were observed in the two groups. Results After treatment, the total effective rate of the observation group was 95.6%, which of the control group was 68.9%, the difference was statistically significant (χ2=5.473, P<0.05). After treatment for 1 day, 3 days, 7 days, the serum bilirubin values of the observation group were (185.6±25.1)μmol/L, (136.7±19.1)μmol/L, (82.1±10.3)μmol/L, respectively, which of the control group were (205.3±26.4)μmol/L, (184.1±20.3)μmol/L, (128.4±16.7)μmol/L, the differences between the two groups were statistically significant (t=3.628, 11.408, 15.829, all P<0.05). The recovery time of serum bilirubin in the observation group was (7.1±2.4)d, which of the control group was (12.9±3.1)d, there was significant difference between the two groups (t=9.924, P<0.05). The hospitalization time of observation group was (3.5±1.1)d, which of the control group was (5.8±1.5)d, the difference between the two groups was statistically significant (t=8.295, P<0.05). Conclusion Live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice is helpful to improve the clinical efficacy, reduce the treatment time and improve the clinical symptoms. Key words: Jaundice neonatal; Phototherapy; Combined bacillus subtilis and enterococcus faecium granules with multivitamines