Protein Containing the GGDEF Domain Affects Motility and Biofilm Formation in Vibrio cholerae and is Negatively Regulated by Fur and HapR.

Objective: This study aimed to investigate whether the VCA0560 gene acts as an active diguanylate cyclase (DGC) in Vibrio cholerae and how its transcription is regulated by Fur and HapR. Methods: The roles of VCA0560 was investigated by utilizing various phenotypic assays, including colony morphological characterization, crystal violet staining, Cyclic di-GMP (c-di-GMP) quantification, and swimming motility assay. The regulation of the VCA0560 gene by Fur and HapR was analyzed by luminescence assay, electrophoretic mobility shift assay, and DNase I footprinting. Results: VCA0560 gene mutation did not affect biofilm formation, motility, and c-di-GMP synthesis in V. cholerae, and its overexpression remarkably enhanced biofilm formation and intracellular c-di-GMP level but reduced motility capacity. The transcription of the VCA0560 gene was directly repressed by Fur and the master quorum sensing regulator HapR. Conclusion: Overexpressed VCA0560 functions as an active DGC in V. cholerae, and its transcription is repressed by Fur and HapR.

Influences of different drying methods on the structural characteristics and prebiotic activity of polysaccharides from bamboo shoot (Chimonobambusa quadrangularis) residues.

In the present study, in order to assess the influences of drying methods on the chemical structures, in vitro digestibility and prebiotic potential of polysaccharides extracted from Chimonobambusa quadrangularis shoot (CPSs), four drying methods, including hot air-, vacuum-, freeze-, and spray-drying, were utilized to dry CPSs. Results revealed that the physicochemical characteristics and prebiotic activity of CPSs varied by different drying methods. In comparison with the other drying methods, freeze-dried CPSs had higher uronic acid content (9.58%), lower medium-high molecular weight (117.63 kDa), smaller particle size (115.97 nm) and better solubility. All four CPSs fractions exhibited low degree of digestibility to pretended human gastric juice (< 2%) and α-amylase (< 5%). The freeze-dried CPSs showed the greatest prebiotic activity as this polysaccharide induced the strongest proliferation effect of probiotic bacteria and the highest production of total short chain fatty acids. Our results collectively provided substantial evidence that the freeze-drying method proposed in this study could be an effective technique in improving the prebiotic potentiality of CPSs.

Comparisons of prebiotic activity of polysaccharides from shoot residues of bamboo (Chimonobambusa quadrangularis) via different ethanol concentrations.

Three polysaccharide fractions from bamboo shoot (Chimonobambusa quadrangularis), CPS70, CPS75, and CPS80, were prepared using a final ethanol concentration of 70%, 75%, and 80% in the precipitation process. In vitro digestibility and the prebiotic activity of CPS70, CPS75, and CPS80 were evaluated and compared. The results indicated that all three of the CPS fractions exhibit a high degree of nondigestibility to human gastric juice (>98.5%) or α-amylase hydrolysis (>94.5%). Compared with the blank control, the three CPS fractions could not only significantly (p < .05) stimulate the proliferation of B. adolescentis, B. infantis, B. bifidum, and L. acidophilus, but also significantly (p < .05) enhance the production of lactic, acetic, propionic, and butyric acids when these polysaccharides were added as alternative carbon sources to glucose during the in vitro fermentation of four probiotics. Furthermore, when comparing the three CPS fractions, CPS75 displayed the strongest prebiotic potential, as this polysaccharide had the strongest effect on the proliferation of probiotic bacteria as well as the greatest effect on SCFAs production. These results demonstrated that the concentration of ethanol used during the precipitation process has a significant impact on the prebiotic activity of CPS. PRACTICAL APPLICATIONS: Ethanol precipitation is the first step when extracting polysaccharides from aqueous extracts as it is simple, rapid, and easy to carry out. This study focuses on how different concentrations of ethanol used in the precipitation process affect the prebiotic potential of bamboo shoot (Chimonobambusa quadrangularis) polysaccharides (CPS). The result indicated that the concentration of ethanol used during the precipitation process has a significant impact on the prebiotic activity of CPS. To our knowledge, it is the first to evaluate the effects of the concentration of ethanol during the process of precipitation on prebiotic potential of polysaccharides, which can subsequently be applied to the optimization of ethanol concentration when precipitating natural polysaccharides for the purpose of in vitro fermentation.

Different Numbers of Long-Pulse 1064-nm Nd-YAG Laser Treatments for Onychomycosis: A Pilot Study.

PURPOSE: To examine the benefits of different numbers of 1064-nm Nd-YAG laser treatments in patients with onychomycosis. METHODS: This was a pilot study of patients with onychomycosis who were divided into three groups: four treatment sessions (group A), eight sessions (group B), and 12 sessions (group C). Only infected nails of degrees II-III (Scoring Clinical Index for Onychomycosis) were included. Treatment was given once a week using a long-pulse Nd-YAG 1064-nm laser. Patients were followed at 8, 16, and 24 weeks after the first treatment. Side effects were recorded. RESULTS: Treatments were completed for 442 nails in 102 patients. The efficacy rates at 8, 16, and 24 weeks were 35.5%, 38.7%, and 37.4% for group A; 31.4%, 41.7%, and 44.0% for group B; and 27.7%, 50.0%, and 55.4% for group C, respectively. There was a significant difference in the efficacy rate at 24 weeks (P = 0.016) between groups A and C, but not for groups A vs. B, or for groups B vs. C. No difference in the efficacy rate at 8 or 16 weeks was observed among the three groups. In all three groups, the efficacy was better for degree II nails than for degree III nails (all P = 0.016) between groups A and C, but not for groups A vs. B, or for groups B vs. C. No difference in the efficacy rate at 8 or 16 weeks was observed among the three groups. In all three groups, the efficacy was better for degree II nails than for degree III nails (all. CONCLUSIONS: The 1064-nm Nd-YAG laser had clinical benefits against onychomycosis. Higher numbers of treatments provided better long-term (24-week) benefits, but had no impact on the short-term outcomes. The efficacy of laser treatment on degree II onychomycosis was better than for degree III.

Effect of stem cell-based therapy for ischemic stroke treatment: A meta-analysis.

Stroke is a major cause of death and long-term disability worldwide. Cell-based therapies improve neural functional recovery in pre-clinical studies, but clinical results require evaluation. We aimed to assess the effects of mesenchymal stem cells on ischemic stroke treatment. We searched the PubMed, Embase and Cochrane databases until July 2015 and selected the controlled trials using mesenchymal stem cells for ischemic stroke treatment compared with cell-free treatment. We assessed the results by meta-analysis using the error matrix approach, and we assessed the association of mesenchymal stem cell counts with treatment effect by dose-response meta-analysis. Seven trials were included. Manhattan plots revealed no obvious advantage of the application of stem cells to treat ischemic stroke. For the comprehensive evaluation index, stem cell treatment did not significantly reduce the mortality of ischemic stroke patients (relative risk (RR) 0.59, 95% confidence interval (CI) 0.29-1.19; ln(RR) 0.54, 95% CI -0.18 to 1.25, p=0.141). The National Institutes of Health Stroke Scale was also not significantly improved by stem cell treatment (standardized mean difference (SMD) 0.94, 95% CI -0.13 to 2.01, p=0.072). The European Stroke Scale was significantly improved using the stem cell treatment (SMD 1.15, 95% CI 0.37-1.92). The dose-response meta-analysis did not reveal a significant linear regression relationship between the number of stem cells and therapeutic effect, except regarding the National Institutes of Health Stroke Scale index. In conclusion, our assessments indicated no significant difference between stem cell and cell-free treatments. Further research is needed to discover more effective stem cell-based therapies for ischemic stroke treatment.

[The impact of interrupted abdominal aorta compression on resuscitation effects of cardiac arrest rabbit].

OBJECTIVE: To determine whether interrupted abdominal aorta compression-cardiopulmonary resuscitation (IAAC-CPR), as a new cardiopulmonary resuscitation (CPR) method, can improve resuscitation effects on rabbits with cardiac arrest (CA). METHODS: Twenty-eight New Zealand rabbits were randomly divided into two groups in equal number (n=14), and they were resuscitated either by standard CPR (STD-CPR, external chest compression) or by IAAC-CPR (abdominal aorta compression at intermission of external chest compression). CA model was reproduced by injection of iced-potassium chloride, and it was maintained for 3 minutes before resuscitation. Electrocardiogram (ECG), hemodynamic indexes were monitored continuously. Aortic systolic pressure and right atrial pressure at baseline and during resuscitation were recorded. Mean arterial pressure (MAP), coronary perfusion pressure (CPP) and recovery of spontaneous circulation (ROSC) indexes were compared between two groups. RESULTS: Compared with STD-CPR group, MAP within 2 minutes of resuscitation (without drugs intervention) in the IAAC-CPR group was significantly increased (30 s: 46.0±12.6 mm Hg vs. 30.0±12.2 mm Hg, 60 s: 57.0±15.6 mm Hg vs. 31.9±9.7 mm Hg, 90 s: 61.9±14.4 mm Hg vs. 36.0±9.8 mm Hg, 120 s: 63.1±12.6 mm Hg vs. 37.8±11.1 mm Hg, all P<0.05), and CPP was also significantly increased (30 s: 29.9±17.4 mm Hg vs. 14.1±11.0 mm Hg, 60 s: 43.1±22.4 mm Hg vs. 14.3±9.8 mm Hg, 90 s: 46.7±20.1 mm Hg vs. 15.9±7.7 mm Hg, 120 s: 50.5±21.0 mm Hg vs. 17.3±9.9 mm Hg, all P<0.05), the time of ROSC was earlier (212.1±43.4 s vs. 307.3±51.2 s, P<0.05), and successful resuscitation rate was significantly higher (85.7% vs. 42.8%, P<0.05). However, there were no statistical differences in ROSC rate and the survivor rate at 24 hours and 48 hours between the two groups, though they were increased. No abdominal organs injury was found in all animals by postmortem study. CONCLUSION: Abdominal aorta compression in the intermittent period of chest compression (IAAC-CPR) increases MAP, CPP and the successful rate of resuscitation, thus improves the outcome.

[Effect of urokinase on cerebral perfusion in rabbits after cardiopulmonary resuscitation].

OBJECTIVE: To investigate the effect of urokinase on cerebral microcirculatory perfusion after cardiopulmonary resuscitation (CPR) in rabbits after cardiac arrest (CA). METHODS: Twenty New Zealand rabbits were assorted into drug thrombolysis group and routine CPR group by randomized digital method with 10 in each group. Potassium chloride injection combined with asphyxia was employed to establish the CA model, CPR and basic life-support were performed in routine CPR group. Following the above treatments, 20 kU/kg urokinase was given in thrombolysis group. The relative cerebral blood flow (rCBF), cerebral blood volume (rCBV) and top teep time (rTTP) were observed by dual-slice spiral CT cerebral perfusion imaging in order to investigate the effect of urokinase on cerebral microcirculatory perfusion in rabbits. RESULTS: rCBF and rCBV in thrombolysis group were significantly higher than those in routine CPR group (rCBF: 1.248±0.139 vs 0.900±0.070, t=6.870, P=0.000; rCBV: 1.206±0.117 vs. 0.969±0.067, t=6.156, P=0.000), and rTTP in thrombolysis group was significantly shorter than that in routine CPR group (0.950±0.037 vs. 1.015±0.026, t=3.777, P=0.004). The cerebral perfusion in thrombolysis group was obviously better than routine CPR group. There were no significant differences in the time for restoration of spontaneous circulation (ROSC), the time for restoration of spontaneous breathing or mean arterial pressure (MAP between routine CPR group and thrombolysis group [ time of ROSC (s): 307.9±96.4 vs. 242.0±71.0, t=-1.741, P=0.099; time of restoration of spontaneous breathing (minutes): 20.6±12.5 vs. 19.3±10.1, t= -0.256, P=0.801; MAP (mm Hg, 1 mm Hg=0.133 kPa): 65.5 = 6.2 vs 69.0±6.7, t=1.217, P=0.239]. CONCLUSION: Thrombolytic therapy with urokinase in CPR could improve the cerebral microcirculatory perfusion in CA rabbits.

Use of rich BHI medium instead of synthetic TMH medium for gene regulation study in Yersinia pestis.

OBJECTIVE: This study is to verify the use of rich BHI medium to substitute synthetic media for gene regulation studies in Yersinia pestis. METHODS: The transcriptional regulation of rovA by PhoP or via temperature upshift, and that of pla by CRP were investigated when Y. pestis was cultured in BHI. After cultivation under 26 °C, and with temperature shifting from 26 to 37 °C, the wild-type (WT) strain or its phoP or crp null mutant (ΔphoP or Δcrp, respectively) was subject to RNA isolation, and then the promoter activity of rovA or pla in the above strains was detected by the primer extension assay. The rovA promoter-proximal region was cloned into the pRW50 containing a promoterless lacZ gene. The recombinant LacZ reporter plasmid was transformed into WT and ΔphoP to measure the promoter activity of rovA in these two strains with the ß-Galactosidase enzyme assay system. RESULTS: When Y. pestis was cultured in BHI, the transcription of rovA was inhibited by PhoP and upon temperature upshift while that of pla was stimulated by CRP. CONCLUSION: The rich BHI medium without the need for modification to be introduced into the relevant stimulating conditions (which are essential to triggering relevant gene regulatory cascades), can be used in lieu of synthetic TMH media to cultivate Y. pestis for gene regulation studies.

Effects of subdiaphragmatic cardiac compression on cardiac arrest during liver transplantation.

Cardiac arrest during upper abdominal surgery such as liver transplantation is a rare but very severe complication. Traditional external cardiac compression has been the mainstay of basic life support in general circumstances. Subdiaphragmatic cardiac compression (SDCC), with no incision in the diaphragm, may be a more effective measure. This maneuver can provide more effective and timely cardiac compression via the already open abdomen in surgery and not add extra trauma. This method can provide a quicker and more effective means of circulation support for intraoperative cardiac arrest patients without adding new injuries. Five cases are reported and all the patients had return of spontaneous circulation (ROSC). This is the first report of the SDCC method.

[An experimental study on the effects of rhythmic abdominal lifting and compression during cardiopulmonary resuscitation in a swine model of asphyxia].

OBJECTIVE: To compare the hemodynamic and respiratory influences of chest compression- cardiopulmonary resuscitation (CC-CPR) and rhythmic abdominal lifting and compression-cardiopulmonary resuscitation (ALC-CPR) in a swine model of asphyxia cardiac arrest (CA), and evaluate the effectiveness of rhythmic abdominal lifting and compression. METHODS: Thirty swines were randomly divided into two groups, with 15 swines in each group. CA model was reproduced by asphyxia as a result of clamping the trachea, and CC-CPR and ALC-CPR was conducted in two groups, respectively. Electrocardiogram (ECG), pulse oxygen saturation [SpO(2)], end-tidal partial pressure of carbon dioxide [P(ET)CO(2)], aorta systolic blood pressure (SBP), diastolic blood pressure (DBP), central venous pressure (CVP), and tidal volume (VT) were monitored continuously from 10 minutes before asphyxia to the end of experiment. The aorta mean arterial pressure (MAP), coronary perfusion pressure (CPP) and minute ventilation (MV) were calculated. Artery blood samples were collected to determine the blood gas analysis at 10 minutes before asphyxia, 10 minutes after asphyxia, and 5, 10, 20 minutes after resuscitation. The restoration of spontaneous circulation (ROSC) rate, 24-hour survival rate and 24-hour neurological function score were observed. RESULTS: There were no significant differences in all mentioned indexes between two groups at 10 minutes before and 10 minutes after asphyxia. At 2 minutes after the resuscitation, the MAP (mm Hg, 1 mm Hg = 0.133 kPa) and CPP (mm Hg) in CC-CPR group were significantly higher than those in ALC-CPR group (MAP: 43.60 ± 12.91 vs. 33.40 ± 6.59, P < 0.05; CPP: 21.67 ± 11.28 vs. 11.80 ± 4.16, P < 0.01), the VT (ml) and MV (L/min)in ALC-CPR group were significantly higher than those in CC-CPR group (VT: 111.67 ± 18.12 vs. 56.60 ± 7.76; MV: 11.17 ± 1.81 vs. 5.54 ± 0.79, both P < 0.01). At 5, 10, 20 minutes after resuscitation, in ALC-CPR group, pH value, arterial partial pressure of oxygen [PaO(2), mm Hg] and arterial oxygen saturation [SaO(2)] were increased, and HCO(3)(-) (mmol/L) and base excess (BE, mmol/L) were decreased, which significantly higher than those in CC-CPR group [pH at 20 minutes after resuscitation: 7.16 ± 0.16 vs. 7.01 ± 0.14; PaO(2): 82.73 ± 13.20 vs. 58.33 ± 17.77; HCO(3)(-): 27.71 ± 3.11 vs. 21.04 ± 3.62; BE: -4.78 ± 4.30 vs. -10.23 ± 2.12; SaO(2): 0.893 ± 0.088 vs. 0.764 ± 0.122], and arterial partial pressure of carbon dioxide [PaCO(2), mm Hg], K(+) (mmol/L) and lactic acid (Lac,mmol/L) were significantly lower than those in CC-CPR group [PaCO(2) at 20 minutes after resuscitation: 49.40 ± 15.60 vs. 79.80 ± 15.35; K(+): 7.18 ± 1.76 vs. 8.55 ± 1.02; Lac: 8.17 ± 1.46 vs. 10.39 ± 1.92], with statistical significant (P < 0.05 or P < 0.01). But the ROSC rate and 24-hour survival rate in ALC-CPR group were significantly higher than those in CC-CPR group (ROSC rate: 80.0% vs. 26.7%, P < 0.01; 24-hour survival rate: 60.0% vs. 13.3%, P < 0.05), and the 24-hour neurological function score was significantly lower than that in CC-CPR group (1.11 ± 0.33 vs. 3.50 ± 0.70, P < 0.01). CONCLUSION: In the incipient stage of cardiopulmonary resuscitation of the swine CA model of asphyxia, compared with CC-CPR, ALC-CPR can be more effective.