Objective:To evaluate the effect of arctigenin (ARG) on cognitive dysfunction induced by sevoflurane anesthesia in aged rats and the role of autophagy-mediated pyroptosis.Methods:Fifty SPF male Sprague-Dawley rats, aged 18-20 months, weighing 550-600 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (C group), sevoflurane anesthesia group (Sev group), sevoflurane anesthesia+ ARG group (Sev+ ARG group), sevoflurane anesthesia+ autophagy inducer rapamycin group (Sev+ RAPA group), and sevoflurane anesthesia+ ARG+ autophagy inhibitor 3-methyladenine (3-MA) group (Sev+ ARG+ 3-MA group). Except for group C, the rats in the other groups inhaled 6% sevoflurane for 3 h to establish the cognitive impairment model. At 30 min before anesthesia, ARG 20 mg/kg was intraperitoneally injected in Sev+ ARG group, rapamycin 7.5 mg/kg was intraperitoneally injected in Sev+ RAPA group, and ARG 20 mg/kg (dissolved in dimethyl sulfoxide) was intraperitoneally injected, followed by immediate intraperitoneal injection of 3-MA 1.5 mg/kg in Sev+ ARG+ 3-MA group. The equal volume of dimethyl sulfoxide was intraperitoneally injected in C group and Sev group. The Morris water maze test was conducted to assess the cognitive function at 48 h after the end of administration. After completion of the Morris water maze test, the hippocampal tissue was taken under deep anesthesia for observation of the pathological changes (after HE staining) which were scored and for determination of neuronal pyroptosis (after propidium iodide staining) and expression of neuronal autophagy-related proteins (microtubule-associated protein 1 light chain 3 [LC3], Beclin-1, p62), pyroptosis-related proteins (NOD-like receptor protein 3 [NLRP3], apoptosis-associated speck-like protein containing a CARD [ASC], pro-cysteine aspartate-specific protease 1 [pro-caspase-1], cleaved-caspase-1, gasdermin D [GSDMD], and N-terminal fragment of gasdermin D [GSDMD-N], interleukin-1β [IL-1β] and IL-18). Results:Compared with C group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the hippocampal injury score and neuronal pyroptosis rate were increased, LC3Ⅱ/LC3Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was up-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were increased in Sev group ( P<0.05). Compared with Sev group, the escape latency was significantly shortened, the time of staying at the original platform quadrant was prolonged, the number of crossing the original platform was increased, the hippocampal injury score and neuronal pyroptosis rate were decreased, LC3Ⅱ/LC3Ⅰ ratio was increased, the expression of Beclin-1 was up-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was down-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were decreased in Sev+ ARG group and Sev+ RAPA group ( P<0.05). Compared with Sev+ ARG group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the hippocampal injury score and neuronal pyroptosis rate were increased, LC3Ⅱ/LC3Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was up-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were increased in Sev+ ARG+ 3-MA group ( P<0.05). Conclusions:AGR can alleviate sevoflurane anesthesia-induced cognitive dysfunction in aged rats, and the mechanism is related to reduction of autophagy-mediated cell pyroptosis.

Objective:To evaluate the effect of arctigenin (ARG) on cognitive dysfunction induced by sevoflurane anesthesia in aged rats and the role of autophagy-mediated pyroptosis.Methods:Fifty SPF male Sprague-Dawley rats, aged 18-20 months, weighing 550-600 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (C group), sevoflurane anesthesia group (Sev group), sevoflurane anesthesia+ ARG group (Sev+ ARG group), sevoflurane anesthesia+ autophagy inducer rapamycin group (Sev+ RAPA group), and sevoflurane anesthesia+ ARG+ autophagy inhibitor 3-methyladenine (3-MA) group (Sev+ ARG+ 3-MA group). Except for group C, the rats in the other groups inhaled 6% sevoflurane for 3 h to establish the cognitive impairment model. At 30 min before anesthesia, ARG 20 mg/kg was intraperitoneally injected in Sev+ ARG group, rapamycin 7.5 mg/kg was intraperitoneally injected in Sev+ RAPA group, and ARG 20 mg/kg (dissolved in dimethyl sulfoxide) was intraperitoneally injected, followed by immediate intraperitoneal injection of 3-MA 1.5 mg/kg in Sev+ ARG+ 3-MA group. The equal volume of dimethyl sulfoxide was intraperitoneally injected in C group and Sev group. The Morris water maze test was conducted to assess the cognitive function at 48 h after the end of administration. After completion of the Morris water maze test, the hippocampal tissue was taken under deep anesthesia for observation of the pathological changes (after HE staining) which were scored and for determination of neuronal pyroptosis (after propidium iodide staining) and expression of neuronal autophagy-related proteins (microtubule-associated protein 1 light chain 3 [LC3], Beclin-1, p62), pyroptosis-related proteins (NOD-like receptor protein 3 [NLRP3], apoptosis-associated speck-like protein containing a CARD [ASC], pro-cysteine aspartate-specific protease 1 [pro-caspase-1], cleaved-caspase-1, gasdermin D [GSDMD], and N-terminal fragment of gasdermin D [GSDMD-N], interleukin-1β [IL-1β] and IL-18). Results:Compared with C group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the hippocampal injury score and neuronal pyroptosis rate were increased, LC3Ⅱ/LC3Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was up-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were increased in Sev group ( P<0.05). Compared with Sev group, the escape latency was significantly shortened, the time of staying at the original platform quadrant was prolonged, the number of crossing the original platform was increased, the hippocampal injury score and neuronal pyroptosis rate were decreased, LC3Ⅱ/LC3Ⅰ ratio was increased, the expression of Beclin-1 was up-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was down-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were decreased in Sev+ ARG group and Sev+ RAPA group ( P<0.05). Compared with Sev+ ARG group, the escape latency was significantly prolonged, the time of staying at the original platform quadrant was shortened, the number of crossing the original platform was reduced, the hippocampal injury score and neuronal pyroptosis rate were increased, LC3Ⅱ/LC3Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, the expression of p62, NLRP3, ASC, IL-1β and IL-18 was up-regulated, and the cleaved-caspase-1/pro-caspase-1 ratio and GSDMD-N/GSDMD ratio were increased in Sev+ ARG+ 3-MA group ( P<0.05). Conclusions:AGR can alleviate sevoflurane anesthesia-induced cognitive dysfunction in aged rats, and the mechanism is related to reduction of autophagy-mediated cell pyroptosis.

Objective To investigate the pharmacodynamic effects of Poria triterpenes on tumor suppression in mice with ascites tumors,and to detect and identify the chemical components of Poria triterpenes using HPLC-LTQ-Orbitrap method,and to explore the potential mechanism of action of Poria triterpenes on tumor suppression in mice with ascites tumors based on both of them using non-labeled definitive proteomics techniques.Methods Poria triterpene parts were extracted by solvent method,and the main components were detected and identified by HPLC-LTQ-Orbitrap liquid mass spectrometer;mice were randomly divided into model group,Poria triterpene group and positive control group(mushroom polysaccharide group),after 21 days of continuous administration,to establish After continuous administration for 21 days,the H22 ascites tumor mouse model was established,and the effect of each drug group on the amount of ascites,spleen index,thymus index,liver index,serum TNF-Alpha and IL-2 in H22 ascites tumor mice was observed after one week of continued administration;then H22 cells were extracted from the ascites of mice,and the H22 cells in the model group and Poria triterpene group were detected by non-lable-free quantitative proteomics technique.The differentially expressed proteins(DEPs)were screened out by using GO,KEGG and other analyses.Results The abdominal water volume in the mouse Poria triterpene group was significantly lower than that in the model group(P<0.05);the thymus index in the mouse Poria triterpene group was significantly higher than that in the model group(P<0.05);the serum levels of TNF-Alpha in the mouse Poria triterpene group were significantly different from those in the model group(P<0.01),and the serum levels of IL-2 in the mouse Poria triterpene group were significantly different from those in the model group(P<0.01).Through the analysis of the chemical composition of Poria triterpene parts,a total of 19 triterpenoids were identified,with four main structural types.A total of 188 differentially expressed proteins were identified in the Poria triterpene group compared with the model group,of which 86 differentially expressed proteins were up-regulated and 102 differentially expressed proteins were down-regulated;GO database analysis showed that the differentially expressed proteins in the Poria triterpene group were mainly involved in the regulation of interleukin-1 production The KEGG database analysis showed that the differentially expressed proteins in the Poria triterpene group were involved in signaling pathways closely related to tumor,mainly MAPK,Apoptosis,mTOR,Wnt and p53 pathways,etc.The genes coding for the seven representative differential proteins were validated at the mRNA level by RT-qPCR.Conclusion The pharmacodynamic study found that Poria triterpenes had tumor suppressive effect on H22 ascites tumor mice,then by proteomics found Poria triterpenes group ascites H22 cell protein compared to the model group changed significantly,the study thus showed that Poria triterpenes for mice ascites tumor mechanism of tumor suppressive effect mainly involves apoptosis,inflammatory response and immune process.

Objective To evaluate the efficacy of endotracheal tube (ETT) with injecting medicine for continuous airway topical anesthesia in inhibiting responses to an indwelling ETT after operation in the patients undergoing maxillofacial surgery. Methods Sixty patients of both sexes, aged 18-64 yr, weig-hing 45-75 kg, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, scheduled for elective maxillofacial surgery under general anesthesia, were divided to control group ( group C) and ETT with in-jecting medicine for continuous airway topical anesthesia group (group E) by a random number table meth-od, with 30 patents in each group. ETT was indwelled until the patients were admitted to intensive care unit (ICU) after the end of surgery, midazolam 0. 02-0. 03 mg∕kg was intravenously injected, and sufentanil was intravenously infused at a rate of 0. 02-0. 10 μg·kg-1·h-1for sedation. In group E, 2% lidocaine 2 ml was injected through the ETT with injecting medicine every 2 h to perform airway topical anesthesia. The equal volume of normal saline was injected through the ETT with injecting medicine every 2 h to maintain the Ramsay sedation score ≥2 in group C. The occurrence of hypertension, tachycardia and bucking and seda-tive requirements were recorded within 24 h after admission to ICU. The concentrations of norepinephrine and methoxyepinephrine in urine were determined by enzyme-linked immunosorbent assay at 24 h after ad- mission to ICU. Results Compared with group C, the frequency of hypertension, tachycardia and bucking were significantly decreased, the consumption of midazolam and sufentanil was reduced, and the concentra-tions of norepinephrine and methoxyepinephrine in urine were decreased in group E (P＜0. 05). Conclusion ETT with injecting medicine can markedly inhibit responses to an indwelling ETT after operation when used for continuous airway topical anesthesia in the patients undergoing maxillofacial surgery.

Objective To compare the toxicity and anti-inflammatory effect of total saponins from Panax japonicus by different extraction technology. Methods The total saponins of sample 1, sample 2, sample 3, sample 4, sample 5 and sample 6 was prepared respectively by different process, and RAW264.7 cells were treated with the samples of different concentration. Then cells morphology was observed under microscope, thiazolyl blue (MTT) method was used to detect cell activity, the nitric oxide (NO) release of RAW264.7 cells was detected with NO kit. Results The cell toxicity of different samples from low to high was as follows:sample 4sample 5>sample 2>sample 6>sample 1>sample 3. Conclusion Among these six different kinds of extraction process of total saponins from Panax japonicus, the total saponins extract by foam fractionation has not only the minimal toxicity, but also the best primary anti-inflammatory effect.

Background:From April to July in 2009 and 2010,unexplained severe hemorrhagic fever-like illnesses occurred in farmers from the Huaiyangshan mountains range.Methods:Clinical specimens (blood,urine,feces,and throat swabs) from suspected patients were obtained and stored.Mosquitoes and ticks in affected regions were collected.Virus was isolated from 2 patients and characterized by whole genome sequencing.Virus detection in additional patients and arthropods was done by virus-specific reverse transcription (RT) PCR.Clinical and epidemiological data of RT-PCR confirmed patients were analyzed.Results:An unknown virus was isolated from blood of two patients and from Haemaphysalis ticks collected from dogs.Whole genome sequence analysis identified the virus as a novel member of the family Bunyaviridae,most closely related to the viruses of the genus Phlebovirus within which it forms a separate lineage.Subsequently,infection was confirmed by RT-PCR in 33 of 58 suspected patients.The illness in these patients was characterized by fever,severe malaise,nausea,vomiting,and diarrhea.Prominent laboratory findings included low white cell- and platelet counts,coagulation disturbances,and elevation of liver enzymes.Hemorrhagic complications were observed in 3 cases,5 (15%) patients died.Conclusions:A novel tick-borne Bunyavirus causing life-threatening hemorrhagic fever in humans has emerged in the Huaiyangshan mountain areas of China.Further studies are needed to determine the epidemiology,geographic distribution and vertebrate animal ecology of this virus.