Analysis of risk factors, pathogenic bacteria characteristics, and drug resistance of postoperative surgical site infection in adults with limb fractures.

PURPOSE: We carried out the study aiming to explore and analyze the risk factors, the distribution of pathogenic bacteria, and their antibiotic-resistance characteristics influencing the occurrence of surgical site infection (SSI), to provide valuable assistance for reducing the incidence of SSI after traumatic fracture surgery. METHODS: A retrospective case-control study enrolling 3978 participants from January 2015 to December 2019 receiving surgical treatment for traumatic fractures was conducted at Tangdu Hospital of Air Force Medical University. Baseline data, demographic characteristics, lifestyles, variables related to surgical treatment, and pathogen culture were harvested and analyzed. Univariate analyses and multivariate logistic regression analyses were used to reveal the independent risk factors of SSI. A bacterial distribution histogram and drug-sensitive heat map were drawn to describe the pathogenic characteristics. RESULTS: Included 3978 patients 138 of them developed SSI with an incidence rate of 3.47% postoperatively. By logistic regression analysis, we found that variables such as gender (males) (odds ratio (OR) = 2.012, 95% confidence interval (CI): 1.235 - 3.278, p = 0.005), diabetes mellitus (OR = 5.848, 95% CI: 3.513 - 9.736, p < 0.001), hypoproteinemia (OR = 3.400, 95% CI: 1.280 - 9.031, p = 0.014), underlying disease (OR = 5.398, 95% CI: 2.343 - 12.438, p < 0.001), hormonotherapy (OR = 11.718, 95% CI: 6.269 - 21.903, p < 0.001), open fracture (OR = 29.377, 95% CI: 9.944 - 86.784, p < 0.001), and intraoperative transfusion (OR = 2.664, 95% CI: 1.572 - 4.515, p < 0.001) were independent risk factors for SSI, while, aged over 59 years (OR = 0.132, 95% CI: 0.059 - 0.296, p < 0.001), prophylactic antibiotics use (OR = 0.082, 95% CI: 0.042 - 0.164, p < 0.001) and vacuum sealing drainage use (OR = 0.036, 95% CI: 0.010 - 0.129, p < 0.001) were protective factors. Pathogens results showed that 301 strains of 38 species of bacteria were harvested, among which 178 (59.1%) strains were Gram-positive bacteria, and 123 (40.9%) strains were Gram-negative bacteria. Staphylococcus aureus (108, 60.7%) and Enterobacter cloacae (38, 30.9%) accounted for the largest proportion. The susceptibility of Gram-positive bacteria to Vancomycin and Linezolid was almost 100%. The susceptibility of Gram-negative bacteria to Imipenem, Amikacin, and Meropenem exceeded 73%. CONCLUSION: Orthopedic surgeons need to develop appropriate surgical plans based on the risk factors and protective factors associated with postoperative SSI to reduce its occurrence. Meanwhile, it is recommended to strengthen blood glucose control in the early stage of admission and for surgeons to be cautious and scientific when choosing antibiotic therapy in clinical practice.

Dynamic changes in intestinal microbiota and metabolite composition of pre-weaned beef calves.

Gut microbes and their metabolites are essential for maintaining host health and production. The intestinal microflora of pre-weaned calves gradually tends to mature with growth and development and has high plasticity, but few studies have explored the dynamic changes of intestinal microbiota and metabolites in pre-weaned beef calves. In this study, we tracked the dynamics of faecal microbiota in 13 new-born calves by 16S rRNA gene sequencing and analysed changes in faecal amino acid levels using metabolomics. Calves were divided into the relatively high average daily gain group (HA) and the relatively low average daily gain group (LA) for comparison. The results demonstrated that the alpha diversity of the faecal microbiota increased with calf growth and development. The abundance of Porphyromonadaceae bacterium DJF B175 increased in the HA group, while that of Lactobacillus reuteri decreased. The results of the LEfSe analysis showed that the microbiota of faeces of HA calves at eight weeks of age was enriched with P. bacterium DJF B175, while Escherichia coli and L. reuteri were enriched in the microbiota of faeces of LA calves. Besides, the total amino acid concentration decreased significantly in the eighth week compared with that in the first week (P < 0.05). Overall, even under the same management conditions, microorganisms and their metabolites interact to play different dynamic regulatory roles. Our results provide new insights into changes in the gut microbiota and metabolites of pre-weaned calves.

Soil microbial residue characteristics in Pinus massoniana lamb. Plantations.

A large amount of stable soil organic matter (SOM) is derived from microbial necromass, which can be assessed by quantifying amino sugar biomarkers. Pinus massoniana Lamb. Plantations are widely distributed in China and play a vital role in forest carbon sequestration. However, the patterns of soil microbial residue remain poorly understood. In this study, amino sugars were used to characterize patterns of soil microbial residues at three soil depths (0-10, 10-20, and 20-30 cm) in P. massoniana plantations of different ages (young, middle-aged, near-mature, mature, and over-mature; denoted as YG, MD, NM, MT, and OM, respectively). In the topsoil (0-10 cm), the total nitrogen (TN) content of the OM forest was the highest, whereas the soil organic carbon (SOC) content of the MT forest was the highest. Consistent with changes in SOC and TN, total microbial residue content decreased with increasing soil depth. However, the total microbial residues C to SOC contribution increased considerably with increasing depth, suggesting that more SOC was derived from microbial residues in the subsoil than that from the topsoil. The fungal residue C to SOC contribution was higher than that of bacterial residue C. Total amino sugar content in the topsoil increased with increasing age, and MT and OM had a significantly higher content than that of other forests. At all soil depths, SOC and TN content predominantly determined microbial necromass, whereas soil microbial biomass content predominantly determined microbial necromass in the topsoil; soil pH predominantly determined microbial necromass in the 10-20 cm soil layer; and soil pH and Ca2+ content were the primary factors in the soil layer below 20 cm. The study provides valuable insights into controls of microbial-derived organic C could be applied in Earth system studies for predicting SOC dynamics in forests.

At Least 10-fold Higher Lubricity of Molecularly Thin D2O vs H2O Films at Single-Layer Graphene-Mica Interfaces.

Interfacial water is a widespread lubricant down to the nanometer scale. We investigate the lubricities of molecularly thin H2O and D2O films confined between mica and graphene, via the relaxation of initially applied strain in graphene employing Raman spectroscopy. Surprisingly, the D2O films are at least 1 order of magnitude more lubricant than H2O films, despite the similar bulk viscosities of the two liquids. We propose a mechanism based on the known selective permeation of protons vs deuterons through graphene. Permeated protons and left behind hydroxides may form ion pairs clamping across the graphene sheet and thereby hindering the graphene from sliding on the water layer. This explains the lower lubricity but also the hindering diffusivity of the water layer, which yields a high effective viscosity in accordance with findings in dewetting experiments. Our work elucidates an unexpected effect and provides clues to the behavior of graphene on hydrous surfaces.

Myristoleic acid produced by enterococci reduces obesity through brown adipose tissue activation.

OBJECTIVE: Dietary fibre has beneficial effects on energy metabolism, and the majority of studies have focused on short-chain fatty acids produced by gut microbiota. Ginseng has been reported to aid in body weight management, however, its mechanism of action is not yet clear. In this study, we focused on the potential modulating effect of ginseng on gut microbiota, aiming to identify specific strains and their metabolites, especially long-chain fatty acids (LCFA), which mediate the anti-obesity effects of ginseng. DESIGN: Db/db mice were gavaged with ginseng extract (GE) and the effects of GE on gut microbiota were evaluated using 16S rDNA-based high throughput sequencing. To confirm the candidate fatty acids, untargeted metabolomics analyses of the serum and medium samples were performed. RESULTS: We demonstrated that GE can induce Enterococcus faecalis, which can produce an unsaturated LCFA, myristoleic acid (MA). Our results indicate that E. faecalis and its metabolite MA can reduce adiposity by brown adipose tissue (BAT) activation and beige fat formation. In addition, the gene of E. faecalis encoding Acyl-CoA thioesterases (ACOTs) exhibited the biosynthetic potential to synthesise MA, as knockdown (KD) of the ACOT gene by CRISPR-dCas9 significantly reduced MA production. Furthermore, exogenous treatment with KD E. faecalis could not reproduce the beneficial effects of wild type E. faecalis, which work by augmenting the circulating MA levels. CONCLUSIONS: Our results demonstrated that the gut microbiota-LCFA-BAT axis plays an important role in host metabolism, which may provide a strategic advantage for the next generation of anti-obesity drug development.

Exosomal miRNAs Profile in Children's Nonalcoholic Fatty Liver Disease and the Correlation with Transaminase and Uric Acid.

OBJECTIVE: The incidence of non-alcoholic fatty liver disease (NAFLD) in children increased rapidly. However, the pathogenesis of NAFLD, especially how non-alcoholic fatty liver progress to non-alcoholic steatohepatitis, is still unclear. This study aims to explore the exosomal miRNAs profiles and the underline pathogenesis of child NAFLD. METHODS: Twenty NAFLD and 20 health control were enrolled in this study. Circulating exosomes were isolated, and RNA sequencing was performed in test set (3 NAFLD/3 Controls). The differentially expressed miRNAs (DEM) were further validated in validation set (17 NAFLD/17 Controls). Spearman correlation -analysis was used to investigate the association between DEM and clinical parameters. RESULTS: Eighty-two miRNAs were differentially expressed (absolute fold change >2 and p < 0.05) in the 2 groups, they were involved in fat acid metabolism, starch and sucrose metabolism, bile acid metabolism and inflammation. miRNA122-5p, miRNA34a-5p, -miRNA155-5p and miRNA146b-3p were up-regulated in NAFLD group (p < 0.05) and positively correlated with body mass index (r, 0.41-0.59), alanine aminotransferase (r, 0.36-0.52), aspartate transaminase (r, 0.31-0.48) and uric acid (UA, r, 0.51-0.69; p < 0.05). CONCLUSIONS: Circulating exosomal miRNAs may be involved in the pathogenesis of NAFLD and correlated with transaminase and UA.

Dysregulated Response of Follicular Helper T Cells to Hepatitis B Surface Antigen Promotes HBV Persistence in Mice and Associates With Outcomes of Patients.

BACKGROUND & AIMS: Production of neutralizing antibodies against hepatitis B surface antigen (HBsAg) is dysregulated in patients with persistent hepatitis B virus (HBV) infection. We investigated mechanisms by which this immune response to the virus is disrupted and whether it can be restored to promote clearance of HBV. METHODS: Immune-competent C57BL/6N and C57BL/6J, as well as mice deficient in follicular helper T cells (Tfh-cell-deficient), B cells, or Foxp3+ T-regulatory cells (Treg cell deficient), were given hydrodynamic injections of pAAV/HBV1.2 plasmids. Some mice were given injections of sorted Tfh cells, pan-B cells, Treg cells, or a blocking antibody against CTLA4. Production of antibodies against HBsAg and clearance of HBV were assessed by flow cytometry, enzyme-linked immunosorbent assay, polymerase chain reaction, and immunohistochemical analyses. We obtained blood samples from patients with HBV infection and isolated Treg cells. We measured the ability of Treg cells to suppress production of interleukin 21 (IL21) in CD4+ T cells. RESULTS: Immune-competent C57BL/6N and C57BL/6J mice transfected with the plasmid encoding HBV had features of viral clearance and viral persistence observed in humans. A Tfh-cell response to HBsAg was required for clearance of HBV and was suppressed by Treg cells in mice with persistent HBV infection. Depletion of Treg cells or inhibition of Treg-cell function (with blocking antibody against CTLA4) restored the Tfh-cell response against HBsAg and clearance of HBV in mice. Impaired Tfh-cell response to HBsAg was observed in blood from patients with chronic HBV infection, responsiveness was restored by depletion of Treg cells or blocking antibody against CTLA4. CONCLUSIONS: In studies of HBV-infected mice and blood from patients with chronic HBV infection, we found a Tfh-cell response to HBsAg of to be required for HBV clearance, and that this response was blocked by Treg cells. Inhibiting Treg-cell activity using neutralizing antibody against CTLA4 restored the ability of Tfh cells to clear HBV infection; this approach might be developed for treatment of patients with chronic HBV infection.

ß-Cell function in obese children and adolescents with metabolic syndrome compared to isolated obesity.

OBJECTIVE: To evaluate ß-cell function in obese children and adolescents meeting clinical criteria for isolated obesity (iOB), isolated components of dysmetabolism (cMD), or metabolic syndrome (MS), and in obese children and adolescents with normal glucose tolerance (NGT), impaired glucose regulation (IGR), or type 2 diabetes (T2DM). STUDY DESIGN: We undertook a prospective study of Han Chinese children and adolescents aged 8-16 years (median 11 ± 1.4) seen in an obesity clinic between May 2013 and 2018. Patients were classified as iOB (53), cMD (139), and MS (139) groups based on clinical criteria. The same patients were also classified as NGT (212), IGR (111), or T2DM (8) based on results of an oral glucose tolerance test (OGTT). The MS patients were classified as NGT [MS](59) and IGR [MS](72) for the further study. All participants also completed a mixed-meal tolerance test (MMTT). RESULTS: Compared with the iOB group, the MS group had significantly higher area under the curve of C-peptide up to the 2 hours (AUC CP) (P = .03) and peak C-peptide (P = .03), adjusted for BMI, age and Tanner stage, on MMTT. However, there was no difference in the insulinogenic index (ΔI30/ΔG30) or oral disposition index (oDI) derived from the OGTT among the three groups. However, 52% of participants with MS had IGR, compared to 28% in the cMD group. Compared with the NGT group, the individuals with IGR had significantly lower ΔI30/ΔG30 (P = .001) and oDI (P < .001). Compared with the iOB group, the NGT[MS] had significantly higher AUC CP (P = .004), peak C-peptide (P = .004) and ΔI30/ΔG30 (P = .007) adjusted for age, but no difference in oDI. Compared with the NGT[MS], the IGR[MS] had significantly lower ΔI30/ΔG30 (P = .005) and oDI (P < .001), but the AUC CP and peak C-peptide had no difference. CONCLUSION: Although the MS youth have ß-cell hyperfunction as a whole, ß-cell dysfunction is present in the early stages of dysmetabolism in obese youth with cMD or MS and worsened across the spectrum from iOB to cMD and MS, contributing to development of T2DM.

Effects of dietary energy and crude protein levels on growth performance, blood profiles, and nutrient digestibility in weaning pigs.

OBJECTIVE: This experiment was conducted to investigate the effect of reducing dietary metabolic energy (ME) and crude protein (CP) levels on growth performance, blood profiles, and nutrient digestibility in weaning pigs. METHODS: A total of 240 crossbred pigs (Duroc×[Landrace×Yorkshire]) with an average body weight of 8.67±1.13 kg were used for a 6-week feeding trial. Experimental pigs were allotted to a 2×3 factorial arrangement using a randomized complete block design. The first factor was two levels of dietary ME density (low ME level, 13.40 MJ/kg or high ME level, 13.82 MJ/kg) and the second factor was three dietary CP levels based on subdivision of early and late weaning phases (low CP level, 19.7%/16.9%; middle CP level, 21.7%/18.9%; or high CP level, 23.7%/20.9%). RESULTS: Over the entire experimental period, there were no significant difference in body weight among groups, but a decrease in diet energy level was associated with an increase in average daily feed intake (p = 0.02) and decrease in gain-feed ratio (G:F) ratio (p<0.01). Decreased CP levels in the diet were associated with a linear increase in average daily gain (p< 0.05) and quadratic increase in G:F ratio (p<0.05). In the early weaning period, blood urea nitrogen concentration tended to increase when ME in the diet decreased and decrease when CP level in the diet decreased (p = 0.09, p<0.01, respectively). Total protein concentration tended to increase when CP level was reduced (p = 0.08). In the late weaning period, blood urea nitrogen concentration decreased linearly as CP level decreased (p<0.01). The CP and crude fat digestibility decreased when ME was decreased by 0.42 MJ/kg (p = 0.05, p = 0.01, respectively). The CP digestibility increased linearly as CP level decreased (p = 0.01). CONCLUSION: A weaning pig diet containing high ME level (13.82 MJ/kg) and low CP level (19.7%/16.9%) can improve pig growth performance and nutrient digestibility.

Inhibitory effects of Danhong Injection and its major constituents on human cytochrome P450 enzymes in vitro.

Danhong Injection (DHI) as a Chinese patent medicine is mainly used to treat ischemic encephalopathy and coronary heart disease in combination with other chemotherapy. However, the information on DHI's potential drug interactions is limited. The goal of this work was to examine the potential P450-mediated metabolism drug interaction arising from DHI and its active components. The results showed that DHI inhibited CYP2C19, CYP2D6, CYP3A4, CYP2E1 and CYP2C9 with IC50 values of 1.26, 1.42, 1.63, 1.10 and 1.67% (v/v), respectively. Danshensu and rosmarinic acid inhibited CYP2E1 and CYP2C9 with IC50 values of 36.63 and 75.76 µm, and 34.42 and 76.89 µm, respectively. Salvianolic acid A and B inhibited CYP2D6, CYP2E1 and CYP2C9 with IC50 values of 33.79, 21.64 and 31.94 µm, and 45.47, 13.52 and 24.15 µm, respectively. The study provides some useful information for safe and effective use of DHI in clinical practice.

Metabolite profiling of ginsenosides in rat plasma, urine and feces by LC-MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract.

Panax ginseng is widely consumed as a functional food in the form of tea, powder, capsules, among others, and possesses a range of pharmacological activities including adaptogenic, immune-modulatory, anti-tumor, anti-aging and anti-inflammatory effects. The aim of this study was to identify and quantify the major ginsenosides and their metabolites in rat plasma, urine and feces after administration of P. ginseng extract using LC-MS/MS. We collected rat plasma samples at 0.5, 1, 2, 4, 8, 12, 24 and 48 h, and the amounts of urine and fecal samples accumulated in 24 h. Fourteen major ginsenosides and their metabolites were observed in fecal samples at high levels; however, low levels of 11 ginsenosides were detected in urine samples. The pharmacokinetics of the major ginsenosides and their metabolites was investigated in plasma. The results indicated that the maximum plasma concentration, time to maximum concentration and area under the curve of compound K were significantly greater than those of other ginsenosides. This study thus provides valuable information for drug development and clinical application of P. ginseng.

Effects of cashew nut testa levels as an alternative to wheat bran in gestating sow diets.

OBJECTIVE: This study was conducted to evaluate the effects of dietary cashew nut testa (CNT) as an alternative feed ingredient to wheat bran on reproductive performance, litter performance, milk composition, and blood profiles of gestating sows. METHODS: Forth multiparous sows (Yorkshire×Landrace) were fed experimental diets starting at 35 days of pregnancy and an initial average body weight (BW) of 211.53±8.86 kg. Each sow was assigned to a treatment based on BW, backfat thickness (BF) and parity with 10 sows per treatment. Treatments were as follows: i) corn-soybean meal based diet with 6% of wheat bran (C0); ii) basal diet with 2% of CNT and 4% of wheat bran (C2); iii) basal diet with 4% of CNT and 2% of wheat bran (C4); and iv) basal diet with 6% of CNT (C6). RESULTS: There were no statistically significant differences in BW and BF of gestating sows throughout the experimental period. However, changes in BF (p = 0.09) and the daily feed intake of sows (p = 0.09) tended to linearly increase during the lactation period. The weaning to estrus interval (WEI) showed a quadratic response to CNT treatment (p = 0.02), and the C2 diet showed the shortest WEI. Litter birth weight (p = 0.04) and piglet birth weight (p = 0.06) were linearly decreased with increase in CNT. Furthermore, there had no significant differences in piglet weight and litter weight in 21 day. Insulin concentration at day 70 of gestation was linearly reduced with increasing CNT level in diets (p = 0.03). CONCLUSION: When 6% CNT replaced wheat bran in gestating sow diets, there were no negative effects on sow performance, but litter birth weight and piglet birth weight were decreased when CNT level increased in gestating sow diets.

Effects of wheat supplementation levels on growth performance, blood profiles, nutrient digestibility, and pork quality in growing-finishing pigs.

OBJECTIVE: This study was conducted to evaluate various wheat supplementation levels on growth performance, blood profiles, nutrient digestibility, and pork quality in growing-finishing pigs. METHODS: A total of 120 growing pigs ([Yorkshire×Landrace]×Duroc), with an average 27.75± 1.319 kg body weight, were used in growth trial. Pigs were allotted into each treatment by body weight and sex in 4 replicates with 6 pigs per pen in a randomized complete block design. Four-phase feeding programs were used in this experiment. The treatments included the following: i) corn-soybean meal (SBM) - based diet (CON), ii) corn-SBM - based diet+15% of wheat (W15), iii) corn-SBM - based diet+30% of wheat (W30), iv) corn-SBM - based diet+45% of wheat (W45), and 5) corn-SBM-based diet+60% of wheat (W60). RESULTS: There was no significant difference in growth performance among the dietary treatments. However, the gain-to-feed (G:F) ratio tended to increase (quadratic, p<0.08) when the pigs were fed a higher wheat diet during the finishing period. The digestibility of crude ash and fat tended to decrease as the wheat supplementation level increased (p<0.08). The proximate analysis of the longissimus muscle was not affected by the dietary level of wheat. The crude ash content in pork was decreased linearly as the wheat supplementation level increased (p = 0.05). There was no significant difference in the pH level, shear force, water holding capacity, and cooking loss of the pork. In pork and fat, L*, a*, and b* values were not significantly different among dietary treatments. CONCLUSION: Wheat can be supplemented up to 60% in a growing-finishing pig without detrimental effects on growth and pork quality. The G:F ratio tended to improve in the finishing period by wheat inclusion.

Various levels of rapeseed meal in weaning pig diets from weaning to finishing periods.

OBJECTIVE: This experiment was conducted to investigate the influence of rapeseed meal (RSM) supplementation in weaning pig diet on growth performance, blood profile, carcass characteristics and economic analysis on weaning to finishing pigs. METHODS: A total of 120 cross bred ([Yorkshire×Landrace]×Duroc) weaning pigs were allotted to 5 treatments in a randomized complete block design. Each treatment had 4 replications with 6 pigs per pen. Five different levels of RSM (0%, 2%, 4%, 6%, and 8%) were used as dietary treatments. RESULTS: Overall, no treatment showed significant differences in growth performance with increased dietary RSM levels. The concentration of blood urea nitrogen (BUN) decreased as dietary RSM levels increased in 6 weeks (linear response, p<0.01). Total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, triiodothyronine, and thyroxine showed no significant differences, neither were there any significant differences in the immune response (IgG and IgA). As the dietary RSM levels of weaning pig diet were increased, no differences were found among dietary treatments upon performing proximate analyses of the pork after finishing. The influence of RSM supplementation on nutrient digestibility and nitrogen retention were not affected by dietary RSM levels either. With increased dietary RSM levels in the weaning pig diet, no differences among dietary treatments were found after performing proximate analyses of the pork's physiochemical properties. In addition, there were no significant differences observed in pork colors, pH levels, and economic benefits. CONCLUSION: Consequently, this experiment demonstrated that weaning pig's diet containing RSM influenced BUN concentration, but there were no detrimental effects on the growth performance of weaning pigs with up to 8% RSM in the diet.

Accuracy of Magnetically Controlled Capsule Endoscopy, Compared With Conventional Gastroscopy, in Detection of Gastric Diseases.

BACKGROUND & AIMS: Diseases of the stomach, including gastric cancer and peptic ulcer, are the most common digestive diseases. It is impossible to visualize the entire stomach with the passive capsule currently used in practice because of the large size of the gastric cavity. A magnetically controlled capsule endoscopy (MCE) system has been designed to explore the stomach. We performed a prospective study to compare the accuracy of detection of gastric focal lesions by MCE vs conventional gastroscopy (the standard method). METHODS: We performed a multicenter blinded study comparing MCE with conventional gastroscopy in 350 patients (mean age, 46.6 y), with upper abdominal complaints scheduled to undergo gastroscopy at a tertiary center in China from August 2014 through December 2014. All patients underwent MCE, followed by conventional gastroscopy 2 hours later, without sedation. We calculated the sensitivity, specificity, positive predictive value, and negative predictive value of detection of gastric focal lesions by MCE, using gastroscopy as the standard. RESULTS: MCE detected gastric focal lesions in the whole stomach with 90.4% sensitivity (95% confidence interval [CI], 84.7%-96.1%), 94.7% specificity (95% CI, 91.9%-97.5%), a positive predictive value of 87.9% (95% CI, 81.7%-94.0%), a negative predictive value of 95.9% (95% CI, 93.4%-98.4%), and 93.4% accuracy (95% CI, 90.83%-96.02%). MCE detected focal lesions in the upper stomach (cardia, fundus, and body) with 90.2% sensitivity (95% CI, 82.0%-98.4%) and 96.7% specificity (95% CI, 94.4%-98.9%). MCE detected focal lesions in the lower stomach (angulus, antrum, and pylorus) with 90.6% sensitivity (95% CI, 82.7%-98.4%) and 97.9% specificity (95% CI, 96.1%-99.7%). MCE detected 1 advanced gastric carcinoma, 2 malignant lymphomas, and 1 early stage gastric tumor. MCE did not miss any lesions of significance (including tumors or large ulcers). Among the 350 patients, 5 reported 9 adverse events (1.4%) and 335 preferred MCE over gastroscopy (95.7%). CONCLUSIONS: MCE detects focal lesions in the upper and lower stomach with comparable accuracy with conventional gastroscopy. MCE is preferred by almost all patients, compared with gastroscopy, and can be used to screen gastric diseases without sedation. Clinicaltrials.gov number: NCT02219529.

Phycicoccus ginsengisoli sp. nov., isolated from cultivated ginseng soil.

Ginseng-cultivated soil is an excellent habitat for soil-borne bacteria to proliferate. A novel strain, DCY87T, was isolated from ginseng-cultivated soil in Gochang County, Republic of Korea, and subsequently characterized by polyphasic approach. Cells were rod shaped, non-motile, aerobic, Gram-reaction-positive, oxidase-negative and catalase-positive. 16S rRNA gene sequence analysis showed that strain DCY87T shared the highest similarity to 'Phycicoccus ochangensis' L1b-b9 (98.7 %). Closely phylogenetic relatives of strain DCY87T were identified: Phycicoccus ginsenosidimutans BXN5-13T (97.9 %), Phycicoccus soli THG-a14T (97.8 %), Phycicoccus bigeumensis MSL-03T (97.3 %), Phycicoccus cremeus V2M29T (97.3 %), Phycicoccus aerophilus 5516T-20T (97.3 %), Phycicoccus dokdonensis DS-8T (97.3 %) and Phycicoccus jejuensis KSW2-15T (97.1 %). The major polar lipids were classified as phosphatidylinositol and diphosphatidylglycerol. The major cellular fatty acids were composed of iso-C15 : 0, anteiso-C15:0, C17 : 0 and C17 : 1ω8c. The menaquinone was resolved as MK-8(H4). Strain DCY87T contained meso-diaminopimelic acid as diamino acid in the cell-wall peptidoglycan and glucose, xylose and rhamnose in the whole-cell sugar. The genomic DNA G+C content was calculated to be 72.7 mol%. DNA-DNA hybridization value between strain DCY87T and 'P. ochangensis' L1b-b9 was estimated to be 50 %. However, DNA-DNA hybridization value obtained between strain DCY87T and P. ginsenosidimutans BXN5-13T, P. soli THG-a14T and P. bigeumensis MSL-03T was well below 17 %. In general, polyphasic taxonomy demonstrated that DCY87T strain represented a novel species within the genus Phycicoccus. Accordingly, we propose the name Phycicoccus ginsengisoli sp. nov. The type strain is DCY87T (=KCTC 39635T=JCM 31016T).

Biotransformation of gypenoside XVII to compound K by a recombinant ß-glucosidase.

OBJECTIVE: To study the ß-glucosidase gene (bgy1) from Lactobacillus brevis that was cloned and expressed in Escherichia coli BL21 (DE3) and then using it for the biotransformation of gypenoside XVII. RESULTS: The bgy1 gene consists of 2283 bp encoding 761 amino acids, with homology to the glycosyl hydrolase family-3 protein domain. The enzyme (Bgy1) hydrolyzed the glucose moieties at the C-3 position and the outer glucose moieties at the C-20 position of gypenoside XVII. Using 0.1 mg enzyme ml(-1) in 20 mM sodium phosphate buffer at 30 °C and pH 6.0, 1 mg gypenoside XVII ml(-1) was transformed into 0.58 mg compound K ml(-1) within 6 h, with a corresponding molar conversion yield of 89 %. CONCLUSION: The recombinant Bgy1 is considered potentially useful for the practical preparation of compound K.

Metabolism profiles of nuciferine in rats using ultrafast liquid chromatography with tandem mass spectrometry.

Nuciferine (NF) is one of the main aporphine alkaloids existing in the traditional Chinese medicine Folium Nelumbinis (lotus leaves). Modern pharmacological studies have demonstrated that NF has a broad spectrum of bioactivities, such as anti-HIV and anti-hyperlipidemic effects, and has been recommended as a leading compound for new drug development. However, the metabolites and biotransformation pathway of NF in vivo have not yet been comprehensively investigated. The present study was performed to identify the metabolites of NF for exploring in vivo fates. Rat plasma and urine samples were collected after oral administration and prepared by liquid-liquid extraction with ethyl acetate. A method based on ultrafast liquid chromatography with tandem mass spectrometry was applied to identify the metabolites. Q1 (first quadrupole) full scan combined with a multiple reaction monitoring (MRM) survey scan were used for the detection of metabolites. MRM-information-dependent acquisition of enhanced product ions was used for the structural identification of detected metabolites. A total of 10 metabolites were identified, including phase I (demethylation, oxidation and dehydrogenation) and phase II (glucuronidation, sulfation and glutathione) biotransformation products. Demethylation is the main metabolic pathway of NF in the body. These results can help in improving understanding of the disposition and pharmacological mechanism of NF in the body. Copyright © 2016 John Wiley & Sons, Ltd.

Percutaneous Intraluminal Radiofrequency Ablation for Malignant Extrahepatic Biliary Obstruction: A Safe and Feasible Method.

BACKGROUD: The Habib™ EndoHBP catheter is a novel bipolar radiofrequency catheter developed for intraluminal ablation to relieve malignant extrahepatic biliary obstruction. Clinical experience with its use is limited and scattered. AIM: The purpose of this study was to evaluate the clinical feasibility and safety of this technique. METHODS: A single central retrospective analysis was performed with patients who underwent percutaneous intraluminal radiofrequency ablation (RFA) combined with biliary stenting for treatment of extrahepatic obstructive jaundice between September 2011 and May 2014. A Habib™ EndoHBP catheter was used for RFA. Clinical and telephonic follow-ups were carried out. Procedure-related complications, stent patency, patient survival rate and postoperative biochemical tests were investigated. RESULTS: All the 47 patients tolerated well a total of 65 RFA procedures with self-expandable metal stents placed. The predominant disease was distal cholangiocarcinoma (16 of 47 cases). No procedure-related hemobilia or infections occurred. The main postablation complication was pain which could be controlled by analgesics. One patient suffered abdominal hemorrhage, diagnosed by blood test and abdominal ultrasonography and cured with conservative therapy. Significantly decreased TBIL and DBIL levels (P < 0.05) were observed on day 7 postoperatively. Stent patency was 149 days (15-281). Median survival was 181 days (15-495) from the time of the first RFA in each patient. CONCLUSIONS: Percutaneous intraluminal RFA combined with biliary stenting is a safe and feasible therapeutic option for unresectable extrahepatic malignant biliary obstruction. Multiple central prospective controlled trials are necessary for the long-term benefits of RFA.

Characterization of biochemical properties of Bacillus subtilis RecQ helicase.

RecQ family helicases function as safeguards of the genome. Unlike Escherichia coli, the Gram-positive Bacillus subtilis bacterium possesses two RecQ-like homologues, RecQ[Bs] and RecS, which are required for the repair of DNA double-strand breaks. RecQ[Bs] also binds to the forked DNA to ensure a smooth progression of the cell cycle. Here we present the first biochemical analysis of recombinant RecQ[Bs]. RecQ[Bs] binds weakly to single-stranded DNA (ssDNA) and blunt-ended double-stranded DNA (dsDNA) but strongly to forked dsDNA. The protein exhibits a DNA-stimulated ATPase activity and ATP- and Mg(2+)-dependent DNA helicase activity with a 3' → 5' polarity. Molecular modeling shows that RecQ[Bs] shares high sequence and structure similarity with E. coli RecQ. Surprisingly, RecQ[Bs] resembles the truncated Saccharomyces cerevisiae Sgs1 and human RecQ helicases more than RecQ[Ec] with regard to its enzymatic activities. Specifically, RecQ[Bs] unwinds forked dsDNA and DNA duplexes with a 3'-overhang but is inactive on blunt-ended dsDNA and 5'-overhung duplexes. Interestingly, RecQ[Bs] unwinds blunt-ended DNA with structural features, including nicks, gaps, 5'-flaps, Kappa joints, synthetic replication forks, and Holliday junctions. We discuss these findings in the context of RecQ[Bs]'s possible functions in preserving genomic stability.