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OBJECTIVE：To study the distri bution and targeting characteristics of Apigenin nanosuspension （AP-NPs）in mice. METHODS：AP-NPs was prepared with ultrasound microprecipitation. Kunming mice were randomly divided into apigenin （AP） solution group and AP-NPs suspension group ，with 45 mice in each group. The mice were given relevant medicine intragastrically （80 mg/kg）；blood sample of eyeball 500 μL were collected before medication（0 h）and 0.25，0.5，1，2，4，6，8，10 h after medication. After the last blood collection ，the mice were sacrificed and their heart ，liver，spleen，lung，kidney and brain tissues were taken. After protein precipitation with methanol ，HPLC method was adopted for determining plasma and tissues. The determination was performed on Shimadzu ODS-SP column with mobile phase consisted of methanol-water （70 ∶ 30，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 340 nm，and column temperature was 35 ℃. The sample size was 20 μL. The concentration of AP in different samples was calculated according to standard curve，main pharmacokinetic parameters （AUC，cmax）of AP and the ratio of peak concentration （ce），relative uptake rate （RUE），uptake ratio and its change value were calculated with DAS 2.0 software and Excel 2010 software；the tissue distribution and targeting characteristics of AP were analyzed. RESULTS：The linear range of AP in plasma and tissue s were 0.1-25.0 μg/mL（all r＞0.99）；the lower limits of quantification were 0.1 μg/mL. RSDs of intra-day and inter-day were all lower than 15％，and the accuracy were 94.37％-117.48％. The extraction recovery rates were all more than 80％. Compared with AP solution group ，the concentrations of AP in plasma sample （during 0.5-6 h），liver tissue （during 0.25-8 h），spleen tissue （during 0.25-8 h）and cerebral tissue （during 0.25-4 h）were increased significantly in AP-NPs suspension group （P＜0.05 or P＜0.01），and the highest in liver tissue. The concentrations of AP in heart tiusse （6 h），liver tissue （10 h），lung tissue （0.5 h），spleen tissue （during 0.25-10 h）were decreased significantly （P＜ 0.05 or P＜0.01）. There was statistical significance in AUC and cmax of AP in plasma and tissue samples between 2 groups（P＜ 0.05）. The ce，RUE，uptake ratio and its change value of liver tissue were the highest ，being 1.34±0.40，1.99±0.29，48.49％ and 15.71％ . CONCLUSIONS ：After AP is made into nanosus- pension，the distribution of drug tissue is changed ，especially targeting effect on liver tissue is improved.
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Objective To investigate the effects of fecal microbiota transplantation on septic gut flora and the cortex cholinergic anti-inflammatory pathway in rats. Methods Sixty clean grade male Sprague-Dawley (SD) rats were divided into normal saline (NS) control group, sepsis model group and fecal microbiota transplantation group by random number table, with 20 rats in each group. The rat model of sepsis was reproduced by injection of 10 mg/kg lipopolysaccharide (LPS) via tail vein, the rats in the NS control group was given the same amount of NS. The rats in the fecal microbiota transplantation group received nasogastric infusion of feces from healthy donor on the 1st day, 2 mL each time, for 3 times a day, the other two groups were given equal dose of NS by gavage. Fecal samples were collected on the 7th day after modeling, the levels of intestinal microbiota composition was determined using the 16SrDNA gene sequencing technology. The brain function was evaluated by electroencephalogram (EEG), and the proportion of each waveform in EEG was calculated. After sacrifice of rats, the brain tissues were harvested, the levels of protein expression of α7 nicotinic acetylcholine receptor (α7nAChR) were determined by Western Blot, and positive cells of Iba-1 in brain tissue were detected by immunohistochemistry method. The levels of interleukins (IL-6 and IL-1β) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Results Seven days after the reproduction of the model, all rats in the NS control group survived, while 10 rats and 8 rats died in the sepsis model group and fecal microbiota transplantation group, respectively, with mortality rates of 50% and 40% respectively. Finally, there were 20 rats in the NS control group, 10 in the sepsis model group and 12 in the fecal microbiota transplantation group. Compared with the NS control group, the diversity and composition of intestinal flora were changed, the incidence of abnormal EEG increased significantly, the expression of α7nAchR in the cortex decreased significantly, and the levels of Iba-1, TNF-α, IL-6 and IL-1β were significantly increased in the model group, suggested that the intestinal flora was dysbiosis, and severe inflammatory reaction occurred in the cerebral cortex, and brain function was impaired. Compared with the model group, the diversity of intestinal flora in the fecal microbiota transplantation group was significantly increased (species index: 510.24±58.76 vs. 282.50±47.42, Chao1 index: 852.75±25.24 vs. 705.50±46.50, both P < 0.05), the dysbiosis of intestinal flora at phylum, family, genus level induced by LPS were also significantly reversed, and with the improvement of intestinal flora, the incidence of abnormal EEG waveforms was lower in the fecal microbiota transplantation group compared with that in the model group [25.0% (3/12) vs. 80.0% (8/10), P < 0.05], and the expression of α7nAChR protein in the cerebral cortex was significantly increased (α7nAChR/β-actin: 1.56±0.05 vs. 0.82±0.07, P < 0.05), immunohistochemistry analysis showed that Iba-1 positive expression of microglia decreased significantly, and cerebral cortex TNF-α, IL-6, IL-1β levels were significantly decreased [TNF-α (ng/L): 6.28±0.61 vs. 12.02±0.54, IL-6 (ng/L): 28.26±3.15 vs. 60.58±4.62, IL-1β (ng/L): 33.63±3.48 vs. 72.56±2.25, all P < 0.05]. Conclusion The results reveal that fecal microbiota transplantation has remarkably modulated the dysbiosis of intestinal microbiota and activated cholinergic anti-inflammatory pathway, and ameliorate the brain dysfunction in septic rats.
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Objective@#To investigate the effects of fecal microbiota transplantation on septic gut flora and the cortex cholinergic anti-inflammatory pathway in rats.@*Methods@#Sixty clean grade male Sprague-Dawley (SD) rats were divided into normal saline (NS) control group, sepsis model group and fecal microbiota transplantation group by random number table, with 20 rats in each group. The rat model of sepsis was reproduced by injection of 10 mg/kg lipopolysaccharide (LPS) via tail vein, the rats in the NS control group was given the same amount of NS. The rats in the fecal microbiota transplantation group received nasogastric infusion of feces from healthy donor on the 1st day, 2 mL each time, for 3 times a day, the other two groups were given equal dose of NS by gavage. Fecal samples were collected on the 7th day after modeling, the levels of intestinal microbiota composition was determined using the 16SrDNA gene sequencing technology. The brain function was evaluated by electroencephalogram (EEG), and the proportion of each waveform in EEG was calculated. After sacrifice of rats, the brain tissues were harvested, the levels of protein expression of α7 nicotinic acetylcholine receptor (α7nAChR) were determined by Western Blot, and positive cells of Iba-1 in brain tissue were detected by immunohistochemistry method. The levels of interleukins (IL-6 and IL-1β) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA).@*Results@#Seven days after the reproduction of the model, all rats in the NS control group survived, while 10 rats and 8 rats died in the sepsis model group and fecal microbiota transplantation group, respectively, with mortality rates of 50% and 40% respectively. Finally, there were 20 rats in the NS control group, 10 in the sepsis model group and 12 in the fecal microbiota transplantation group. Compared with the NS control group, the diversity and composition of intestinal flora were changed, the incidence of abnormal EEG increased significantly, the expression of α7nAchR in the cortex decreased significantly, and the levels of Iba-1, TNF-α, IL-6 and IL-1β were significantly increased in the model group, suggested that the intestinal flora was dysbiosis, and severe inflammatory reaction occurred in the cerebral cortex, and brain function was impaired. Compared with the model group, the diversity of intestinal flora in the fecal microbiota transplantation group was significantly increased (species index: 510.24±58.76 vs. 282.50±47.42, Chao1 index: 852.75±25.24 vs. 705.50±46.50, both P < 0.05), the dysbiosis of intestinal flora at phylum, family, genus level induced by LPS were also significantly reversed, and with the improvement of intestinal flora, the incidence of abnormal EEG waveforms was lower in the fecal microbiota transplantation group compared with that in the model group [25.0% (3/12) vs. 80.0% (8/10), P < 0.05], and the expression of α7nAChR protein in the cerebral cortex was significantly increased (α7nAChR/β-actin: 1.56±0.05 vs. 0.82±0.07, P < 0.05), immunohistochemistry analysis showed that Iba-1 positive expression of microglia decreased significantly, and cerebral cortex TNF-α, IL-6, IL-1β levels were significantly decreased [TNF-α (ng/L): 6.28±0.61 vs. 12.02±0.54, IL-6 (ng/L): 28.26±3.15 vs. 60.58±4.62, IL-1β (ng/L): 33.63±3.48 vs. 72.56±2.25, all P < 0.05].@*Conclusion@#The results reveal that fecal microbiota transplantation has remarkably modulated the dysbiosis of intestinal microbiota and activated cholinergic anti-inflammatory pathway, and ameliorate the brain dysfunction in septic rats.
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Objective To evaluate the efficacy of regional cerebral oxygen saturation(rSO2) combined with neuroelectrophysiological monitoring in guiding intraoperative blood pressure management in elderly patients undergoing carotid endarterectomy. Methods Thirty patients of both sexes, aged 65-80 yr, of American Society of Anesthesiologists physical statusⅡorⅢ, scheduled for elective carotid endar-terectomy, were divided into 2 groups(n=15 each)using a random number table: control group(group C)and rSO2combined with neuroelectrophysiological monitoring group(group M). During occlusion of carotid artery, the vasoactive drugs were used to make systolic blood pressure(SBP)increase by 20%-30% of the baseline value in group C and to make rSO2not less than 20% of the baseline value, the ampli-tude of somatosensory evoked potential P40 not less than 50% of the baseline value and the amplitude of e-lectroencephalogram voltage not less than 50% in group M. SBP and rSO2were recorded immediately after intubation(T1), at 5 min after anesthesia induction(T2), at 5 min after blocking the carotid artery (T3), at 5 min after opening the carotid artery(T4)and immediately after extubation(T5). Decrease in rSO2≥20% of the baseline value was recorded. The carotid artery occlusion time, myocardial oxygen con-sumption and consumption of vasoactive drugs during occlusion were recorded. Results Compared with group C, SBP was significantly decreased at T3, and the consumption of vasoactive drugs and myocardial oxygen consumption were reduced in group M(P<0.05), and no significant change was found in rSO2at each time point in group M(P>0.05). Decrease in rSO2≥20% of the baseline value was not found in two groups. Conclusion rSO2combined with neuroelectrophysiological monitoring provides guidance for intra-operative blood pressure management in patients undergoing carotid endarterectomy.