Analysis and identification of key anti-inflammatory molecules in Eerdun Wurile and exploration of their mechanism of action in microglia.

Ethnomedicine Eerdun Wurile (EW) can significantly promote poststroke neuro-recovery through modulation of microglia polarization. Fraction 4-6 (F4-6) isolated from EW via serial fractionation inhibits the expression of pro-inflammatory genes in LPS stimulated microglia. However, the key active molecules of F4-6 have not been identified. Herein, we identified alantolactone (Ala) and dehydrodiisoeugenol (Deh) as the active anti-inflammatory components of F4-6 by UPLC-qTof MS analysis. We confirmed that, F4-6, Ala, Deh and mixture of Ala and Deh (Mix) downregulate the expression of several pro-inflammatory genes including Ccl2, Cox2 and Il6 in LPS-treated microglia in a similar pattern. At the same time upregulate the expression of anti-inflammatory genes including Hmox1, Tgfß, Igf1 and Creb1. Moreover, the conditioned culture media obtained from F4-6 treated microglia significantly enhanced proliferation of N2a cells, and promoted neurite outgrowth possibly through upregulation of Nefh and Dlg4. Mechanistically, F4-6 strongly downregulated the expression of NF-κB p65, while also inhibiting the nuclear translocation of p65, leading to the suppression of transcription of pro-inflammatory genes initiated by NF-κB. Collectively, our data identified and quantified the key chemicals of EW and provide insights into the optimization of the herbal composition for neuroprotection.

Enhancement of the heterogeneous adsorption and incorporation of uraniumVI caused by the intercalation of ß-cyclodextrin into the green rust.

The influence of intercalated anions on the structure and composition of green rusts supplies a theoretical possibility for the investigation of the structural modification of FeII/FeIII (oxyhydr)oxide materials. ß-Cyclodextrin was intercalated into the mixed-valent iron-based hydroxide layers to synthesize new green rust materials (ß-CD GRs), pursuing high-capacity uraniumVI (UVI) sorption. The molar ratios of FeII to FeIII and the molar ratios of ß-CD GR to FeII + FeIII had a significant effect on the synthesis of ß-CD GRs. The synthesis process was further optimized by the quadric predictor and desirability function in a central composite design in combination. Both strong acidity and alkalinity were harmful to the adsorption of ß-CD GRs towards UVI. The pseudo-first-order kinetic model and Langmuir isotherm model were appropriate in fitting the whole adsorption process. The maximum monolayer adsorption capacity of ß-CD GRs was 2548.61 mg/g. The presence of mimic groundwater constituents explicitly deteriorated the interaction between ß-CD GR and UVI species. Nanoscale nodules and particles were formed on the ß-CD GR after the adsorption experiments. The peaks at 1159 and 609 cm-1 vanished with the band at 1103 cm-1 being left-shifted to 1117 cm-1 in the FTIR spectra of ß-CD GR during the heterogeneous process. The intercalation of ß-CD brought obvious enhancement of UVI species sorption to the GR material, which was combinedly driven by several reaction pathways and different from the unmodified GRs.

Meta-analysis of the efficacy and safety of non-vitamin K antagonist oral anticoagulants with warfarin in Latin American patients with atrial fibrillation.

BACKGROUND: Data of non-vitamin K antagonist oral anticoagulants (NOACs) in current management of atrial fibrillation (AF) are predominantly derived from North American and European regions. However, the effects of NOACs for stroke prevention in Latin America remain unclear. Therefore, we aimed to compare the efficacy and safety of NOACs with warfarin in Latin American patients with AF. METHODS: The PubMed and Embase databases were systematically searched until July 12, 2019 for applicable randomized clinical trials. The risk ratios (RRs) and 95% confidence intervals (CIs) were pooled using a random-effects model. RESULTS: Four trials involving 8943 Latin American patients were included in this meta-analysis. In anticoagulated patients with AF, Latin American patients had higher rates of stroke or systemic embolism and all-cause death compared with non-Latin American subjects. Compared with warfarin use, the use of NOACs was significantly associated with reduced risks of stroke or systemic embolism, major bleeding, intracranial bleeding, and any bleeding in Latin American patients. There were no significant differences in the risks of ischemic stroke, all-cause death, and gastrointestinal bleeding between Latin and non-Latin American groups. All the interactions between Latin and non-Latin American groups about efficacy and safety outcomes of NOACs compared with warfarin were non-significant (all Pinteraction > .05). CONCLUSIONS: Our meta-analysis suggested that the use of NOACs was at least non-inferior to warfarin use for stroke prevention in Latin American patients with AF.

Analysis and Identification of Tumorigenic Targets of MicroRNA in Cancer Cells by Photoreactive Chemical Probes.

Photoactive RNA probes have unique advantages in the identification of microRNA (miR) targets due to their ability for efficient conjugation to the target sequences by covalent crosslinking, providing stable miR-mRNA complexes for further analysis. Here, we report a highly efficient and straightforward method for miR target identification that is based on photo-reactive chemical probes and RNA-seq technology (denotes PCP-Seq). UV reactive probes were prepared by incorporating psoralen in the specific position of the seed sequence of miR. Cancer cells that were transfected with the miR probes were treated with UV, following the isolation of poly(A) RNA and sequencing of the transcriptome. Quantitative analysis of RNA-seq reads and subsequent validation by qPCR, dual luciferase assay as well as western blotting confirmed that PCP-Seq could highly efficiently identify multiple targets of different miRs in the lung cancer cell line, such as targets PTTG1 and PTGR1 of miR-29a and ILF2 of miR-34a. Collectively, our data showed that PCP-Seq is a robust strategy for miR targets identification, and unique in the identification of the targets that escape degradation by miRISC and maintain normal cellular level, although their translation is repressed.

Design of curdlan-based pH-sensitive polymers with endosome buffering functionality for siRNA delivery.

Developing nucleic acid-based tools to control disease-relevant gene expression in human disorders, such as siRNAs, opens up potential opportunities for therapeutics. Because of their high molecular weight and polyanionic nature, synthetic siRNAs fail to cross biological membranes by passive diffusion and therefore, generally require transmembrane siRNA delivery technologies to access the cytoplasm of target cells. To create a biocompatible siRNA delivery agent, we chemically modified natural polysaccharide curdlan derivative 6AC-100 in a regioselective manner to introduce different ratios of imidazole rings in the amino units (denoted as Curimi) and evaluated their siRNA binding ability, cytotoxicity, endosome buffering capacity and siRNA transfection efficiency. The novel curdlan based Curimi polymers formed nanoparticles with siRNA at pH 7.4 in range of 85-105 nm and their size distribution increased along with decreasing pH condition. The zeta potential increased by lowering pH value as well. Curimi polymers showed lower toxicity and higher buffering capacity compared to 6AC-100, and efficiently delivered siRNA against to PLK1 into cancer cells, and subsequently, significantly inhibited target mRNA level. Our result suggested that novel curdlan based Curimi polymers may be used as efficient siRNA carrier for cancer therapy.

[The evaluation of cerebral function by diffusion weighted imaging after norepinephrine-induced hypertensive perfusion therapy in pig model of cardiac arrest].

OBJECTIVE: To evaluate the changes in cerebral function of pigs with cardiac arrest (CA) after recovery of spontaneous circulation (ROSC) after hypertension perfusion therapy induced by norepinephrine (NE). METHODS: Ventricular fibrillation (VF) was induced by electrical stimulation, and standard cardiopulmonary resuscitation (CPR) was performed after VF for 4 minutes. The pigs with successful ROSC were randomly divided into two group, each n=5. The pigs in the hypertensive reperfusion group were given with NE immediately to maintain the mean arterial pressure (MAP) at 130% before VF for 4 hours; MAP of the pigs in normal reperfusion group was maintained for 4 hours as baseline. The changes in hemodynamics were observed for 4 hours in both groups. Cerebral cortex was scanned with diffusion weighted imaging (DWI) before VF and 1 hour and 3 hours after ROSC, and the dynamic changes in brain functional imaging were observed. Twenty-four hours after ROSC, brain biopsy were collected and examined after hematoxylin and eosin staining (HE). RESULTS: Compared with the normal reperfusion group, heart rate (HR), MAP, cardiac output (CO) and coronary perfusion pressure (CPP) in the hypertensive reperfusion group showed a tendency to increase (ROSC 30 minutes HR: 167±8 bpm vs. 140±15 bpm, ROSC 1 hour MAP: 131±9 mm Hg vs. 108±10 mm Hg, ROSC 1 hour CO: 4.9±0.1 L/min vs. 3.4±0.5 L/min, ROSC 2 hours CPP: 118±12 mm Hg vs. 88±1 mm Hg, P<0.05 or P<0.01). There was no obvious abnormality as shown by DWI before and after resuscitation, and the apparent diffusion coefficient (ADC) showed a tendency to decrease after resuscitation in both groups. The ADC in the normal reperfusion group was decreased more than that in the hypertensive reperfusion. Pathological study showed that the protective effect of the hypertensive reperfusion on brain tissue was better than that of the normal reperfusion group. CONCLUSION: Hypertensive reperfusion can produce hemodynamic changes, and an increase in cerebral blood flow, thus it produces a protective effect on brain to promote the recovery of neurological function in pigs with CA after resuscitation.

Effects of hypothermia on brain injury assessed by magnetic resonance imaging after cardiopulmonary resuscitation in a porcine model of cardiac arrest.

OBJECTIVE: To evaluate the effects of hypothermia on cerebral edema and metabolism, a porcine model of cardiac arrest was assessed by magnetic resonance imaging during the first 72 hours after restoration of spontaneous circulation (ROSC). METHODS: Ventricular fibrillation was induced in 33 pigs. After 8 minutes of untreated ventricular fibrillation, 30:2 cardiopulmonary resuscitation was performed. After ROSC, 30 survival animals were randomly divided into normothermia group (n = 15) and hypothermia group (n = 15). The hypothermia group immediately received endovascular cooling to regulate temperature to 33°C, which was maintained for 12 hours, followed by passive rewarming at 0.5°C/h to 37°C. Diffusion-weighted imaging and (1)hydrogen proton magnetic resonance spectroscopy were acquired for each group at 6, 12, 24, and 72 hours after ROSC. RESULTS: Compared with the normothermia group, the hypothermia group exhibited a higher 72-hour survival (73.3% vs. 33.3%, P = .028) and a superior neurological deficit score (P = .031). Cerebral injury was found in both groups, but a lesser decrease in the apparent diffusion coefficient and N-acetyl aspartate/creatinine (P < .05) and a greater increase in choline/creatinine (P < .05) were found in the hypothermia group. CONCLUSIONS: Magnetic resonance imaging could effectively detect the dynamic trend of cerebral injury in a porcine model of cardiac arrest within the first 72 hours after ROSC. Hypothermia produced a protective effect on neurological function by reducing brain edema and formation of adverse metabolites.

Evaluation of respiratory dysfunction in a pig model of severe acute dichlorvos poisoning.

BACKGROUND: Respiratory failure is the main cause of death in acute organophosphorus pesticide poisoning. In this study, a pulse-induced contour cardiac output monitor was used to evaluate the respiratory status in a pig model of acute dichlorvos poisoning. METHODS: Twenty female pigs were randomly allocated to dichlorvos (n = 7), atropine (n = 7), and control (n = 6) groups. In the dichlorvos group, pigs were administered 80% emulsifiable dichlorvos (100 mg/kg) via a gastric tube. In the atropine group, pigs were similarly administered dichlorvos, and 0.5 hours later, atropine was injected to attain and maintain atropinization. The control group was administered saline solution. Arterial blood gas was measured at 0, 0.5, 1, 2, 4, and 6 hours post-injection. The extravascular lung water index and pulmonary vascular permeability index were recorded by the pulse-induced contour cardiac output monitor. At termination of the study, the animals were euthanized, the lung wet-to-dry weight ratio was determined, and histopathology was observed. RESULTS: In the dichlorvos group, the extravascular lung water index and pulmonary vascular permeability index were substantially increased from 0.5 hours and were particularly high within 1 hour. In the atropine group, these indices increased initially, but decreased from the 1-hour mark. The control group exhibited no obvious changes. In both the dichlorvos and atropine groups, the extravascular lung water index was negatively correlated with partial pressure of oxygen/fraction of inspiration oxygen (PO2/FiO2) and positively correlated with the pulmonary vascular permeability index. Compared with the control group, the lung wet-to-dry weight ratio markedly increased and the histopathological findings obviously changed in the dichlorvos group, but only mildly increased and changed, respectively, in the atropine group. CONCLUSION: The extravascular lung water index is an appropriate and valuable parameter for assessment of respiratory function in acute dichlorvos poisoning.

Investigation of myocardial stunning after cardiopulmonary resuscitation in pigs.

OBJECTIVE: To investigate cardiac function and myocardial perfusion during 48 h after cardiopulmonary resuscitation (CPR), further to test myocardial stunning and seek indicators for long-term survival after CPR. METHODS: After 4 min of untreated ventricular fibrillation, fifteen anesthetized pigs were studied at baseline and 2 h, 4 h, 24 h, and 48 h after restoration of spontaneous circulation (ROSC). Hemodynamic data, echocardiography and gated-single photon emission computed tomography myocardial perfusion images were carried out. RESULTS: Mean arterial pressure (MAP), coronary perfusion pressure (CPP) and cardiac troponin I (CTNI) showed significant differences between eventual survival animals and non-survival animals at 4 h after ROSC (109.2 ± 10.7 mmHg vs. 94.8 ± 12.3 mmHg, P=0.048; 100.8 ± 6.9 mmHg vs. 84.4±12.6 mmHg, P=0.011; 1.60 ± 0.13 ug/L vs. 1.75 ± 0.10 ug/L, P=0.046). Mitral valve early-to-late diastolic peak velocity ratio, mitral valve deceleration time recovered 24 h; ejection faction and the summed rest score recovered 48 h after ROSC. CONCLUSION: Cardiac systolic and early active relaxation dysfunctions were reversible within survival animals; cardiac stunning might be potentially adaptive and protective after CPR. The recovery of MAP, CPP, and CTNI could be the indicators for long-term survival after CPR.

Effects of norepinephrine on kidney in a swine model of cardiopulmonary resuscitation.

BACKGROUND: The aim of this study was to study the effects of norepinephrine (NE)-induced hypertension (HT) on renal biochemistry, enzymology, and morphology after restoration of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in swine. METHODS: After 4 minutes of ventricular fibrillation, standard CPR was carried out. The survivors were then divided into 2 groups. The HT group (n = 5) received 0.4 to 1.0 µg kg⁻¹ min⁻¹ of NE continuously to maintain the mean arterial pressure (MAP) at 130% of the baseline (ie, MAP before ventricular fibrillation). The normal pressure (NP) group (n = 5) received 0.2 to 0.5 µg kg⁻¹ min⁻¹ NE continuously to maintain MAP at the baseline level. Hemodynamic status and oxygen metabolism were monitored, and blood urea nitrogen and creatinine were measured in blood samples obtained at baseline and at 10 minutes, 2 and 4 hours after ROSC. At 24 hours after ROSC, the animals were killed and the kidney was removed to determine Na⁺-K⁺-ATPase and Ca²âº-ATPase activities and histologic changes under a light and electron microscopy. RESULTS: mean arterial pressure, cardiac output, and coronary perfusion pressure were significantly higher (P < .01), whereas the oxygen extraction ratio was lower in the HT group than in the NP group (P < .05). Blood urea nitrogen and creatinine increased in the NP group but did not change in the HT group. Renal ATP enzyme activity was significantly higher in the HT group than the NP group (Na⁺-K⁺-ATP enzyme: 4.024 ± 0.740 U versus 3.190 ± 0.789 U, Ca²âº-ATP enzyme: 3.615 ± 0.668 versus 2.630 ± 0.816; both P < .05). The HT group showed less cellular edema, necrosis, and fewer damaged mitochondria compared with the NP group. CONCLUSION: These data suggest that inducing HT by NE helps to maintain stable hemodynamic status and oxygen metabolism and may protect the kidney in terms of biochemistry, enzymology, and histology after CPR.

Evaluation of cerebral metabolism by ¹H-magnetic resonance spectroscopy for 4°C saline-induced therapeutic hypothermia in pig model of cardiac arrest.

BACKGROUND: Previous studies have shown that therapeutic hypothermia could improve neurologic recovery when induced after cardiac arrest, but dynamic changes in cerebral metabolism have not been studied at low temperature. In this study, we aim to evaluate hypothermia-induced changes in pigs' cerebral metabolism by (1)H-magnetic resonance spectroscopy (¹H-MRS). MATERIAL AND METHODS: Ten anesthetized Landrace (25-30 kg) pigs were randomized into 2 groups and subjected to 4 minutes of ventricular fibrillation, followed by cardiopulmonary resuscitation. The hypothermic group was given an infusion of 30 mL/kg of 4°C normal saline (NS) at an infusion rate of 1.33 mL/kg per minute starting after restoration of spontaneous circulation (ROSC), then 10 mL/kg per hour for 4 hours. The control group received the same infusion of room temperature NS. Core temperature and hemodynamic variables were monitored at baseline and repeatedly for 240 minutes after ROSC. The ¹H-MRS scans were obtained at baseline, 1 hour, and 3 hours after successful ROSC to observe the dynamic changes of cerebral metabolism at different temperatures. RESULTS: The mean reduction of temperature was 1.5°C ± 0.4°C in the hypothermic group. There was no difference in hemodynamic variables between groups. ¹H-MRS detected statistically significant (P < .01) changes in cerebral metabolism between the control and hypothermia groups (P < .01). CONCLUSIONS: Infusion of 4°C NS can effectively reduce cerebral metabolism after successful cardiopulmonary resuscitation and have a protective effect on the recovery of neurologic function. The ¹H-MRS technology can be used as a powerful tool to evaluate interventions in the treatment of cardiopulmonary resuscitation.

Effect of continuous compressions and 30:2 cardiopulmonary resuscitation on global ventilation/perfusion values during resuscitation in a porcine model.

OBJECTIVE: Rescue ventilations during bystander resuscitation, although previously considered essential, interrupt the continuity of chest compressions and might have deleterious effects in basic life support. This study was undertaken to analyze the global ventilation/perfusion values of continuous compressions and 30:2 cardiopulmonary resuscitation to determine the effectiveness for each approach in a porcine model of prolonged bystander cardiopulmonary resuscitation for ventricular fibrillation. DESIGN: Prospective, randomized animal study. SETTING: A university animal research laboratory. SUBJECTS: Twenty-four male domestic pigs (n = 12/group) weighing 30 ± 2 kg. INTERVENTIONS: All animals had ventricular fibrillation induced by programmed electrical stimulation instruments and were randomized into two groups. Continuous compressions or 30:2 compression/rescue ventilation cardiopulmonary resuscitation was performed in each group. MEASUREMENTS AND MAIN RESULTS: Continuous respiratory variables, hemodynamic parameters, and blood gas analysis outcomes were recorded, and global ventilation/perfusion values were calculated. Alveolar minute volume and global ventilation/perfusion values decreased progressively after ventricular fibrillation, but cardiac output was stable. The global ventilation/perfusion value was higher in the ventilation cardiopulmonary resuscitation group than that in the continuous compression group (p < .0001) and was higher than normal. Coronary perfusion pressure was progressively decreased after 6 mins of cardiopulmonary resuscitation and greatly fluctuated in the ventilation cardiopulmonary resuscitation group. Coronary perfusion pressure was higher in the continuous compression group than that in the ventilation cardiopulmonary resuscitation group after 9 mins of cardiopulmonary resuscitation (p < .05). Values for pH and Pao2 progressively decreased, but there were no significant differences between the two groups, except for pH at 12 mins of cardiopulmonary resuscitation and Paco2 after 3 mins of cardiopulmonary resuscitation. CONCLUSIONS: In the first 12 mins of cardiopulmonary resuscitation, continuous compressions could maintain relatively better coronary perfusion pressure, Pao2, and global ventilation/perfusion values than 30:2 cardiopulmonary resuscitation. Therefore, rescue ventilation during 12 mins of simulated bystander cardiopulmonary resuscitation did not improve hemodynamics or outcomes compared with compression-only cardiopulmonary resuscitation.

[The influence of hypothermia therapy on enzymology and pathology of lung after cardiac arrest].

OBJECTIVE: To study the effect of hypothermia on pro-inflammatory mediators in serum, the enzymology and pathology of lung tissue. METHODS: Ventricular fibrillation for 4 minutes was induced in 10 domestic pigs. Standard cardiopulmonary resuscitation was given to them. They were then divided into two groups according to the random table after restoration of spontaneous circulation (ROSC): low temperature group (n=5): pigs were given an infusion 30 ml/kg of 4 centigrade normal saline (NS) at an infusion rate of 1.33 ml * kg(-1) * min(-1), started after ROSC for 22 minutes, then 10 ml * kg(-1) * h(-1) for 4 hours; ambient temperature group (n=5) received the same infusion of NS in room temperature. Hemodynamic parameters were observed, blood samples were collected to measure tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in serum before ventricular fibrillation and 10 minutes, 2 hours, 4 hours after ROSC. Na(+)-K(+)-ATPase of lung tissue was determined 24 hours after ROSC, and the pathology and ultrastructure of the lung were studied. RESULTS: There was no significant difference in the hemodynamic parameters, except the temperature, between low temperature and ambient temperature groups. TNF-alpha contents at 10 minutes, 2 hours, 4 hours after ROSC in low temperature group were (15.55+/-1.65), (17.06+/-0.86), (12.52+/-1.82) ng/L, and the IL-6 contents were (173.80+/-15.01), (184.09+/-13.44), (73.17+/-6.95) ng/L, while the TNF-alpha contents at 10 minutes, 2 hours, 4 hours after ROSC in ambient temperature group were (20.09+/-1.32), (26.18+/-1.16), (29.18+/-1.20) ng/L, and the IL-6 contents were (176.92+/-16.68), (239.17+/-13.18), (405.48+/-55.49) ng/L. The pro-inflammatory mediators in low temperature group were reduced significantly (P<0.05 or P<0.01). Low temperature could significantly reduce the activity of Na(+)-K(+)-ATPase [(3.78+/-1.14) U/L vs. (6.22+/-1.23) U/L, P<0.01]. The pathology of lung was milder in low temperature group compared with that of ambient temperature group. CONCLUSION: Hypothermia therapy with infusion of 4 centigrade NS can reduce the release of pro-inflammatory mediators, inhibit the ATPase activity of alveolar membrane, and shows a protective effect on lung tissue against low perfusion.