Effects of probiotic supplementation on 12 min run performance, mood management, body composition and gut microbiota in amateur marathon runners: A double-blind controlled trial.

Background: Probiotic supplementation has a positive effect on endurance exercise performance and body composition in athletes, but the underlying mechanisms remain unclear. Gut microbiota can provide measurable markers of immune function in athletes, and microbial composition analysis may be sensitive enough to detect stress and metabolic disorders caused by exercise. Methods: Nineteen healthy active amateur marathon runners (15 male and 4 female) with a mean age of 29.11 years volunteered to participate in this double-blind controlled study. Based on the performance of the Cooper 12-min running test (CRT), the participants were allocated into two groups to receive either a probiotic formulation comprising lactobacillus acidophilus and bifidobacterium longum (n = 10) or placebo containing maltodextrin (n = 9) for five weeks. Consistency of diet and exercise was ensured throughout the experimental period. Before and after the intervention, all participants were assessed for CRT, emotional stability and gastrointestinal symptoms, gut microbiota composition, body composition and magnetic resonance imaging (MRI) indicators of skeletal muscle microcirculation. Results: Compared to before the intervention, the probiotics group showed an increase in CRT score (2.88 ± 0.57 vs 3.01 ± 0.60 km, P＜0.05), significant improvement in GSRS and GIQLI (9.20 ± 4.64 vs 7.40 ± 3.24, 118.90 ± 12.30 vs 127.50 ± 9.85, P＜0.05), while these indicators remained unchanged in the control group, with a significant time-group interaction effect on gastrointestinal symptoms. Additionally, some MRI metabolic cycling indicators of the thigh skeletal muscle also changed in the probiotics group (P＜0.05). Regarding microbiota abundance, the probiotics group exhibited a significant increase in the abundance of beneficial bacteria and a significant decrease in the abundance of harmful bacteria post-intervention (P＜0.05). Conclusion: As a sports nutritional supplement, probiotics have the potential to improve athletic performance by optimizing the balance of gut microbiota, alleviating gastrointestinal symptoms.

Prediction of half-marathon performance of male recreational marathon runners using nomogram.

BACKGROUND: Long-distance running is a popular competitive sport. We performed the current research as to develop an easily accessible and applicable model to predict half-marathon performance in male recreational half-marathon runners by nomogram. METHODS: Male recreational half-marathon runners in Zhejiang Province, China were recruited. A set of literature-based and panel-reviewed questionnaires were used to assess the epidemiological conditions of the recruited runners. Descriptive and binary regression analyses were done for the profiling and identification of predictors related to higher half-marathon performance (completing time ≤ 105 min). Participants were assigned to the training set (n = 141) and the testing set (n = 61) randomly. A nomogram was used to visually predict the half-marathon performance, and the receiver operating characteristic (ROC) was used to evaluate the predictive ability of the nomogram. RESULTS: A total of 202 participants (median age: 49 years; higher half-marathon performance: 33.7%) were included. After multivariate analysis, three variables remained as significant predictors: longer monthly running distance [adjusted odds ratio (AOR) = 0.992, 95% confidence interval (CI): 0.988 to 0.996, p < 0.001], faster mean training pace (AOR = 2.151, 95% CI: 1.275 to 3.630, p < 0.001), and better sleep quality [the Pittsburgh Sleep Quality Index (PSQI), AOR = 2.390, 95% CI: 1.164 to 4.907, p = 0.018]. The AUC of the training and testing sets in nomogram were 0.750 and 0.743, respectively. Further ternary and linear regression analyses corroborated the primary findings. CONCLUSIONS: This study developed a nomogram with good potential to predict the half-marathon performance of recreational runners. Our results suggest that longer monthly running distance, faster mean training pace and better sleep quality notably contribute to better half-marathon performance.

Impact of Running Exercise on Intervertebral Disc: A Systematic Review.

CONTEXT: Running is one of the most popular sports worldwide. However, controversies exist regarding how running affects runner's intervertebral discs (IVD). OBJECTIVE: The purpose of this study was to systematically review studies that evaluated IVD morphology or composition changes in response to running exercise, to determine the impact of running exercise on IVD. DATA SOURCES: A systematic literature search was performed for 4 major databases: PubMed, Cochrane, Embase, and Web of Science. STUDY SELECTION: Inclusion criteria were as follows: (1) healthy people without known IVD disease or major complications such as tuberculosis (IVD degeneration or low back pain are considered as minor complications); (2) subjects performed 1-time or regular running exercises; (3) pre and post comparison of runners or comparison between runners and healthy control subjects; (4) direct or indirect IVD morphology or composition measured; (5) IVD assessed before and after either acute or chronic running exercise, or compared cross-sectionally between runners and controls. Exclusion criteria were as follows: (1) reviews, editorials, letters or abstracts only; (2) animal studies; (3) subjects performed exercise other than running. STUDY DESIGN: Systematic review. LEVEL OF EVIDENCE: Level 3. DATA EXTRACTION: The extracted data included study design and primary outcomes of the included studies. The Newcastle-Ottawa scale (NOS) was used to evaluate study quality and risk of bias. RESULTS: A total of 13 studies with 632 participants were included in the final analysis; 4 studies measured IVD changes using stature or spinal height, and the other 9 measured IVD changes using magnetic resonance imaging; 6 studies found that running acutely and negatively impacts IVD; 3 out of 5 cross-sectional studies found that IVD parameters are better for runners than controls; 1 longitudinal study found no significant difference in IVD before and after training for marathon in runners; 1 longitudinal study found no significant difference in changes of IVD between runners and controls after 15 years of follow-up. CONCLUSION: Negative changes in IVD exist for a short period of time after running, which may be due to the temporary compression pushing water content out of the disc. Cross-sectional studies suggest that long-term running exerts a mild positive effect on IVD; however, this inference has not been confirmed by high-quality longitudinal studies.

Higher Risk of Sarcopenia in Older Adults with Type 2 Diabetes: NHANES 1999-2018.

INTRODUCTION: Recent studies suggested that sarcopenia may be a significant comorbidity of diabetes mellitus (DM). Nonetheless, studies with nationally representative data are scarce, and the changing trend of sarcopenia prevalence over time is largely unknown. Therefore, we aimed to estimate and compare the prevalence of sarcopenia in diabetic and nondiabetic US older population, and to explore the potential predictors of sarcopenia as well as the trend of sarcopenia prevalent in the past decades. METHODS: Data were retrieved from the National Health and Nutrition Examination Survey (NHANES). Sarcopenia and DM were defined according to corresponding diagnosis criteria. Weighted prevalence was calculated and compared between diabetic and nondiabetic participants. The differences among age and ethnicity groups were explored. RESULTS: A total of 6,381 US adults (>50 years) were involved. The overall prevalence of sarcopenia was 17.8% for US elders, and the prevalence was higher (27.9% vs. 15.7%) in those with diabetes ones than those without. Stepwise regression revealed that sarcopenia was significantly associated with DM (adjusted odds ratio = 1.37, 95% CI: 1.08-1.22; p < 0.05) after controlling for potential confounders including gender, age, ethnicity, educational level, BMI, and muscle strengthening activity. A slight fluctuation but overall increasing trend of sarcopenia prevalence was observed among diabetic elders, while no obvious changing trend was observed in their counterparts in recent decades. CONCLUSION: Diabetic US older adults face significantly higher risk of sarcopenia when compared with their nondiabetic counterparts. Gender, age, ethnicity, educational level, and obesity were important influencing factors of sarcopenia development.

Lysosomal iron recycling in mouse macrophages is dependent upon both LcytB and Steap3 reductases.

Iron that is stored in macrophages as ferritin can be made bioavailable by degrading ferritin in the lysosome and releasing iron back into the cytosol. Iron stored in ferritin is found as Fe3+ and must be reduced to Fe2+ before it can be exported from the lysosome. Here we report that the lysosomal reductase Cyb561a3 (LcytB) and the endosomal reductase six-transmembrane epithelial antigen of prostate 3 (Steap3) act as lysosomal ferrireductases in the mouse macrophage cell line RAW264.7 converting Fe3+ to Fe2+ for iron recycling. We determined that when lysosomes were loaded with horse cationic ferritin, reductions or loss of LcytB or Steap3 using CRISPR/Cas9-mediated knockout technology resulted in decreased lysosomal iron export. Loss of both reductases was additive in decreasing lysosomal iron export. Decreased reductase activity resulted in increased transcripts for iron acquisition proteins DMT1 and transferrin receptor 1 (Tfrc1) suggesting that cells were iron limited. We show that transcript expression of LcytB and Steap3 is decreased in macrophages exposed to Escherichia coli pathogen UTI89, which supports a role for these reductases in regulating iron availability for pathogens. We further show that loss of LcytB and Steap3 in macrophages infected with UTI89 led to increased proliferation of intracellular UTI89 suggesting that the endolysosomal system is retaining Fe3+ that can be used for proliferation of intravesicular pathogens. Together, our findings reveal an important role for both LcytB and Steap3 in macrophage iron recycling and suggest that limiting iron recycling by decreasing expression of endolysosomal reductases is an innate immune response to protect against pathogen proliferation and sepsis.

Prognostic signatures associated with high infiltration of Tregs in bone metastatic prostate cancer.

Metastatic cancer especially bone metastasis (BM) is the lethal end-stage of castration-resistant prostate cancer (CRPC). To understand the possible molecular mechanisms underlying the development of the distant metastasis is of potential clinical value. We sought to identify differentially expressed genes between patient-matched primary and bone metastatic CRPC tumors. Functional enrichment, protein-protein interaction networks, and survival analysis of DEGs were performed. DEGs with a prognostic value considered as candidate genes were evaluated, followed by genetic analysis of tumor infiltrating immune cells based on Wilcoxon test and immunofluorescence identification. Expression profiles analysis showed that 381 overlapping genes were screened as differentially expressed genes (DEGs), of which 16 DEGs were randomly selected to be validated and revealed that most of these genes showed a transcriptional profile similar to that seen in the datasets (Pearson's r = 0.76). Six core genes were found to be involved in regulation of extracellular matrix receptor interaction and chemotactic activity, and four of them were significantly correlated with the survival of PCa patients with bone metastases. Immune infiltration analysis showed that the expressions levels of COL3A1, RAC1, FN1, and SDC2 in CD4+T cells were significantly higher than those in tumor cells, especially regulatory T cell infiltration was significantly increased in BM tumors. We analyzed gene expression signatures specifically associated with the development of bone metastases of CRPC patients. Characterization of genes associated with BM of mCRPC is critical for identification of predictive biomarkers and potential therapeutic targets.

[Study on oral absorption mechanisms of puerarin in nanocrystals self-stabilized pickering emulsion].

Nanocrystals self-stabilized Pickering emulsion(NSSPE) is a new kind of emulsion where only nanocrystals of poorly soluble drugs are used as stabilizers. Our previous study showed that NSSPE with Ligusticum chuanxiong oil as the main oil phase can significantly promote oral absorption of puerarin. The present study aimed to explore its absorption mechanism in oral administration. The in vitro dissolution test was carried out to study the effect of NSSPE on release of puerarin. The effects and mechanism of NSSPE on uptake and transport of puerarin across Caco-2 cell were investigated. The results showed that the drug release rate of NSSPE was similar to that of nanocrystals, with their cumulative dissolution of puerarin not affected by pH of releasing mediums, both significantly higher than that of crude material. The uptake of puerarin in NSSPE was concentration-dependent and significantly higher than that of solution or surfactant stabilized emulsion. Genistein and indomethacin, inhibitors of lipid rafts/caveolin, could significantly reduce the uptake of puerarin in NSSPE. Compared with solution, NSSPE and surfactants stabilized emulsion obviously increased transport rate K_a and apparent permeability coefficient P_(app) of puerarin in AP → BL direction, but there was no significant difference in BL → AP direction. It could be inferred that there were both passive and active transport mechanisms, as well as lipid raft/caveolin mediated endocytosis for absorption of NSSPE. The promoted oral absorption of puerarin in NSSPE was mainly related to the existing nanocrystal form which could promote dissolution, puerarin as well as Ligusticum chuanxiong oil which could promote drug transmembrane transport and inhibit drug efflux. It is the unique structure and composition of the compound NSSPE that promoted the oral absorption of puerarin.

[Prognostic Significance of CUEDC1 in Patients with Acute Myeloid Leukemia (non-M3)].

OBJECTIVE: To investigate the prognostic significance of CUEDC1 in patients with acute myeloid leukemia (non-M3). METHODS: 52 cases newly diagnosed AML (non-M3) were selected and enrolled in AML non-M3 group, at the same time, 10 cases of iron doficiency anemia were selected and enrolled in control group. The bone marrow mononuclear cells(BMMC) were isolated from bone marrow of patients, the expression level of CUEDC1 in BMMC was detected by RT-PCR, the expression level of CUEDC1 mRNA in BMMC of AML-subtype patients was compared. The AML patients were divided into low and high expression groups according to the expression level of CUEDC1 mRNA, and the complete remission rate after the first chemothrapy course was compared, and the relative expression level of CUEDC1 mRNA between the remission and the non-remission group were compared. RESULTS: CUEDC1 was expressed in BMMC of 52 newly diagnosed patients with AML (non-M3) of all subtypes, which was higher than that in control group (P<0.05), and the expression level of CUEDC1 mRNA in M5 patients was the highest (P<0.05). In CUEDC1 low expression group, induced complete remisson rate （76.2%,16/21） after the first course of treatment seemed higher than that of the high expression group（67.7%,21/31）, but the difference was not statistically significant; the expression level of CUEDC1 mRNA in the remission group of patients with newly diagnosed AML(non-M3) was lower than that in the non-remission group(P<0.05). CONCLUSION: CUEDC1 is highly expressed in newly diagnosed patients with AML, among which the CUEDC1 mRNA expression level in M5 patients is the highest, the expression of CUEDC1 mRNA possibly relates to the prognosis of patients with AML.

Association of vitamin E on the risk of ovarian cancer: a meta-analysis.

Many researches were conducted to assess the association of vitamin E intake on the risk of ovarian cancer, with conflict results. The current meta-analysis of published observational studies aimed to investigate the effect of vitamin E intake on ovarian cancer risk. The summary relative risks (RRs) with corresponding 95% confidence intervals (CIs) were calculated to measure the effectiveness of vitamin E intake on ovarian cancer risk using a random-effects model. As a result, 14 studies including 4597 patients were identified. Eleven studies reported about total vitamin E intake, eight studies about vitamin E intake from food only and five studies about vitamin E intake from supplement only on the risk of ovarian cancer. Overall, the summary RRs on ovarian cancer risk was 0.95 (95%CIs = 0.78-1.16) in total vitamin E intake, 0.99 (95%CIs = 0.77-1.27) in vitamin E intake from food only and 0.82 (95%CIs = 0.54-1.25) in vitamin E intake from supplement only. Results in subgroup analyses by study design and geographic location were consistent with overall result. In conclusions, the findings of this meta-analysis suggested that high intake of vitamin E from food or vitamin E supplement had no significant effect on the risk of ovarian cancer.

Astragaloside IV Exerts a Myocardial Protective Effect against Cardiac Hypertrophy in Rats, Partially via Activating the Nrf2/HO-1 Signaling Pathway.

Previous evidence suggested that astragaloside IV (ASIV) had a cardioprotective effect, but the potential mechanisms were undetermined. This study is aimed at validating the prevention of cardiac hypertrophy in chronic heart failure (CHF) rats and hypertrophy in H9c2 cardiomyocytes by ASIV and at exploring the potential mechanism involved. CHF rat models of abdominal aortic constriction (AAC) were used with the aim of determining the protective effect of ASIV in cardiac hypertrophy in the rats. We proved that ASIV could attenuate cardiac hypertrophy by improving left ventricular function and structure and showed that the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2) and its downstream gene heme oxygenase-1 (HO-1) increased in the high-dose ASIV intervention group. To further investigate the specific mechanism of ASIV, we hypothesized that ASIV might prevent cardiac hypertrophy via activating the Nrf2/HO-1 signaling pathway. We established a cardiomyocyte hypertrophy model induced by angiotensin II (Ang II), which was then transfected with Nrf2 shRNA, to knock down the expression of the Nrf2 gene. We found that the protective effect of ASIV against Ang II-induced cardiomyocyte hypertrophy was abolished in the Nrf2 shRNA transfection group, ultimately aggravating cardiomyocyte hypertrophy induced by Ang II, and it is possible that oxidative stress may be involved in this process. Our results demonstrated that ASIV improved cardiac function and inhibited cardiac hypertrophy by upregulating Nrf2, and this effect was partially achieved by stimulating the Nrf2/HO-1 signaling pathway, suggesting that ASIV could have therapeutic potential for the treatment of cardiac hypertrophy and CHF.

[Clinical Significance of TF and VEGF Expressions on Peripheral CD14 Positive Monocytes in Patients with Diffuse Large B Cell Lymphoma].

OBJECTIVE: To investigate the clinical significance of tissue factor (TF) and vascular endothelial growth factor (VEGF) expression on peripheral blood CD14 positive monocytes in patients with diffuse large B cell lymphoma (DLBCL). METHODS: The expressions of TF and VEGF on peripheral CD14+ monocytes in 41 patients with DLBCL (DLBCL group) before chemotherapy and after 4 chemotherapeutic courses, and in 20 healthy subjects (control group) were detected by flow cytometry respectively, meanwhile, the relationship of the expression of TF and VEGF with international prognostic indexes (IPI) and short-term effects were analysed. RESULTS: The expression levels of TF and VEGF on peripheral CD14+ monocytes in DLBCL group were significantly higher than those in control group (P<0.01), and a positive correlation was found between the two groups (r=0.755, P<0.01). The expression of TF and VEGF on CD14+ monocytes in patients with prognostic risk factors significantly increased as compared with those in patients without prognostic risk factors (P<0.05), but there were no significant differences of TF and VEGF expressions on CD14+ monocytes in DLBCL group with different sex, age, subtypes (P＞0.05). As compared with patients without prognostic risk factors, the expression levels of TF and VEGF on CD14+ monocytes of patients with prognostic risk factors significantly increased (P<0.05). The expression of TF and VEGF on CD14+ monocytes in DLBCL group showed an increasing tendency along with the increase of IPI index (P<0.01). The expression levels of TF and VEGF on CD14+ monocytes in remission group before chemotherapy were lower than those in non-remission group (P<0.01); after chemotherapy, the expression levels of TF and VEGF on CD14+ monocytes in remission group were lower than those before chemotherapy (P<0.05), while the TF and VEGF expression levels in non-remission group were no singnificauly different from TF and VEGF levels before chemtherapy (P>0.05), the survival of patients in group with low expression of TF and VEGF was superior to that in group with high expression of TF and VEGF (P<0.05). CONCLUSION: The paripheral blood CD14+ monocytes in DLBCL patients highly express the TF and VEGF, which relate with IPI, therapeutic efficacy and survival, thus the TF and VEGF expression levels are of reference significance for evaluating the therapeutic efficacy and prognosis of patients.

Ferritin in serum and urine: A pilot study.

Serum ferritin reflects total body iron stores, thus a low serum ferritin is used as a parameter of iron deficiency. In healthy adults in Japan, urine ferritin levels were about 5% of serum ferritin levels, with a correlation coefficient of 0.79. It is not known whether a low urine ferritin could serve as a non-invasive screen for iron deficiency. If so, this might be useful for neonates and young children, avoiding phlebotomy to screen for iron deficiency. However, for urinary ferritin screening to be feasible, ferritin must be measurable in the urine and correlate with serum ferritin. Testing should also clarify whether the iron content of ferritin in serum and urine are similar. In this pilot feasibility study we measured ferritin in paired serum and urine samples of healthy adult males, healthy term neonates, growing preterm neonates, and children who had very high serum ferritin levels from liver disorders or iron overload. We detected ferritin in every urine sample, and found a correlation with paired serum ferritin (Spearman correlation coefficient 0.78 of log10-transformed values). These findings suggest merit in further studying urinary ferritin in select populations, as a potential non-invasive screen to assess iron stores.

[Construction of A Lentiviral Vector Carrying CUEDC1 Gene and Its Effect on the Proliferation and Colony-formating Ability of MOLT-4 Cells].

OBJECTIVE: To construct a lentiviral vector carrying human CUEDC1 gene, to establish leukemic cell line MOLT-4 stably expressing recombinant plasmid, to analyze the expression of CUEDC1 in MOLT-4 cells and to investigate its effect on the proliferation of MOLT-4 cells. METHODS: The CUEDC1 gene was amplified by RT-PCR, and then was subcloned into the lentiviral vector pCDH to generate a lentiviral vector pCDH-CUEDC1. Recombinant lentivirus was generated by co-transfection of 3 plasmids, and transfected into MOLT-4 cells. The Real-time PCR and Western blot were respectively applied to detect the expression of CUEDC1 mRNA and protein, the CCK-8 and colony formation assay were used to evaluate the effect of CUEDC1 on proliferation of MOLT-4 cells. RESULTS: The recombinant lentiviral vector pCDH-CUEDC1 had been constructed successfully. After infection of MOLT-4 cells with the lentivirus, the recombinant plasmid could stably up-regulate the expression of CUEDC1 and protein. The CCK-8 detection and colony formation assay showed that exogenous CUEDC1 could significantly promote cell growth and the colony formation of MOLT-4 cells. CONCLUSION: The recombinant lentiviral vector carrying human CUEDC1 has been successfully constructed, exogenous CUEDC1 can significantly promote cell growth and the colony formation of MOLT-4 cells.

[Effect of OCT4A Gene on the Biological Characteristics of K562 Cells].

OBJECTIVE: To evaluate biological effects of OCT4A gene on K562 cells and explore the molecular mechanism of K562 cell apoptosis. METHODS: Two recombinant lentiviral vectors were constructed, which could stablely up- regulate and down- regulate OCT4A protein. Recombinant lentivirus was generated by co-transfection of three-plasmids and transfec-ted into K562 cells. The experiments were divided into 5 groups: normal, pLVX-OCT4A-ZsGreen1, pLVX vector control, PLB-OCT4A shRNA and non-specific shRNA groups. Western blot was applied to detect the expression of OCT4A protein, the cell counting kit-8 was applied to evaluate the effect of OCT4A on proliferation of K562 cells. The apoptosis and differentiation of K562 cells were detected by flow cytometry with AnnexinV/7-AAD double staining. The mRNA expressions of caspase-3,BIM,BCL-xL,BAX in K562 cells were determined by real time PCR. RESULTS: The OCT4A fragment was amplified by reverse transcription polymerase chain reaction(RT-PCR), the 2 lentiviral vectors were successfully constructed. In comparson with those in the control group, the expression of OCT4A protein of pLVX-OCT4A-ZsGreen1 group was significantly increased, but decreased in PLB-OCT4A shRNA group. CCK-8 assay showed that the higher the content of OCT4A protein, the faster the cell proliferation. The apoptosis rate was (3.48±0.52)% of pLVX-OCT4A-ZsGreen1 group, which was lower than that of control group, while the apoptosis rate PLB-OCT4A shRNA group was (7.25±0.57)%, which was higher than that of control group (P<0.05), however, the K562 cells differentiation was not influenced(P>0.05). Compared with control group, the gene expression of Caspase-3,BIM and BAX was down-regulated(P>0.05), but a significant up-regulation of BCL-xL gene expression was observed(P<0.05). CONCLUSION: Two lentiviral vectors have been successfully constructed, which can stably up- and down- regulate the expression of OCT4A in K562 cells respectively. OCT4A can promote the K562 cell proliferation and inhibit the apoptosis, the mechanism may be related with up-regulation of BCL-xl expression.

Evaluation of truncated G protein delivered by live attenuated Salmonella as a vaccine against respiratory syncytial virus.

Human respiratory syncytial virus (RSV) can cause severe acute lower respiratory tract disease leading to numerous hospitalizations and deaths in the infant and elderly populations worldwide, while no vaccine or effective drug is available for RSV infections. In the present study, truncated G protein was successfully expressed both in prokaryotic and eukaryotic system, and high levels of serum IgG in response to truncated G protein were observed both in GD-protein group (intramuscularly with purified GD protein) and GD-VNP20009 group (challenged via the oral route with 1 × 109 CFU of pLIVE-RSV-GD-VNP20009 strains) since 21th day, and GD-VNP20009 significantly reduced the productions of IL-1ß, IL-6, and TNF-α, histamine and pathological features caused by the RSV Long strain (P < .01). Our data indicated that Salmonella typhimurium can be used to deliver truncated G DNA vaccine and represents a promising effect to protect host against RSV.

Effective dispersion of multi-walled carbon nanotubes in aqueous solution using an ionic-gemini dispersant.

A promising ionic-gemini molecule, 4, 4'-di (n-tetradecyl) diphenylmethane disulfate salt (DSDM), is reported for effective dispersion of multi-walled carbon nanotubes (MWCNTs) in aqueous medium in the present investigation. The dispersibility and stability of the DSDM-modified MWCNTs were characterized by UV-vis spectrophotometer, Raman spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy and zeta potential measurements. The hydrophobic interaction between alky chains and carbon nanotubes as well as the π-π-stacking interaction between benzene rings and carboatomic rings in MWCNTs enables a successful modification of DSDM onto the MWCNT surface. The dispersed MWCNTs individually existed in dispersion with no structural damage, indicating a much better performance than the MWCNTs dispersed by the sodium dodecylbenzene sulfonate (SDBS), a frequently reported single-chain ionic dispersant. Surface potential measurements showed that the DSDM-modified MWCNTs were negatively charged, giving rise to electrostatic repulsion between the MWCNTs in aqueous solution. A better MWCNT dispersion effect was observed with the increase in MWCNT surface potential, and the dispersion with high MWCNT surface potential presents high dispersion stability with no agglomeration appeared for more than 5 months. The magnesium (Mg) matrix composite fabricated based on the DSDM-dispersed MWCNTs demonstrated excellent mechanical properties compared to pure Mg. Our research may provide an alternative way to improve the mechanical properties of composites.

Therapeutic effect of herb residue fermentation supernatant on spleen­deficient mice.

To minimize the waste of active ingredients present in herb residues, in the present study, probiotics of Bacillus subtilis, Aspergillus oryzae and Lactobacillus plantarum M3 were selected to reuse herb residues from Jianweixiaoshi tablets, and the therapeutic effects of the herb residue fermentation supernatant were evaluated using a spleen­deficient mouse model. The results of the present study indicated that the fermentation supernatant may effectively improve the immunity of mice, as measured by body weight, spleen and thymus index, and inflammatory cytokines, including interleukin (IL)­2, IL­4 and interferon­Î³. The viable cell count and denaturing gradient gel electrophoresis results indicated that the fermentation supernatant markedly enhanced bacterial diversity and the number of lactobacilli in mouse intestines. Therefore, the combination of the Jianweixiaoshi herb residue and probiotics provided a novel method to reuse herb residues and may in the future contribute to the treatment of spleen deficiency.

Evaluation of the accuracy and sensitivity of high­throughput sequencing technology using known microbiota.

Next generation sequencing provides an excellent platform to explore microbiota in any given environment, and little work is required to evaluate the accuracy and sensitivity of high­throughput sequencing technology. In the present study, a known microbiota containing Escherichia coli, Lactobacillus plantarum, Streptococcus thermophilus, Bifidobacterium bifidum, Bacillus subtilis, Enterococcus faecalis and Salmonella typhimurium was used to evaluate the high­throughput sequencing technology. The results suggested that there were 122.7 operational taxonomic units (OTUs) in all groups, which is 17.5­fold (the whole OTU number/the actual bacterial number) greater compared with the actual microbial number in each group, and the Venn method indicated that only 46.38% (64/138), 58.70% (81/138), 86.13% (118/137), 83.57% (117/140) and 89.29% (125/140) of the common OTUs were identified in groups A, B, C, D and E, of which the majority of OTUs did not belong to known bacteria. In addition, the DNA extraction and amplification efficiency failed to identify bacteria at the phylum, class, order, family, genus and species levels, which may further increase false information of microbial analysis. In conclusion, the present study provided basic datato investigate the potential drawbacks of high­throughput sequencing technology, which will help researchers to avoid exaggerating the bacterial number when this technology is applied to study microbiota in particular environments.

High­throughput sequencing analyses of oral microbial diversity in healthy people and patients with dental caries and periodontal disease.

Recurrence of oral diseases caused by antibiotics has brought about an urgent requirement to explore the oral microbial diversity in the human oral cavity. In the present study, the high­throughput sequencing method was adopted to compare the microbial diversity of healthy people and oral patients and sequence analysis was performed by UPARSE software package. The Venn results indicated that a mean of 315 operational taxonomic units (OTUs) was obtained, and 73, 64, 53, 19 and 18 common OTUs belonging to Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria, respectively, were identified in healthy people. Moreover, the reduction of Firmicutes and the increase of Proteobacteria in the children group, and the increase of Firmicutes and the reduction of Proteobacteria in the youth and adult groups, indicated that the age bracket and oral disease had largely influenced the tooth development and microbial development in the oral cavity. In addition, the traditional 'pathogenic bacteria' of Firmicutes, Proteobacteria and Bacteroidetes (accounted for >95% of the total sequencing number in each group) indicated that the 'harmful' bacteria may exert beneficial effects on oral health. Therefore, the data will provide certain clues for curing some oral diseases by the strategy of adjusting the disturbed microbial compositions in oral disease to healthy level.