ObjectiveTo investigate the mechanism by which Anmei Dan （AMD） regulates recombinant sirtuin 3/nuclear factor erythroid 2-related factor 2（SIRT3/Nrf2） signaling to ameliorate mitochondrial oxidative stress and improve the cognitive function in aged sleep-deprived mice. MethodsSixty aged C57 mice were randomly assigned to a control group， a model group， a melatonin （1.3 mg·kg^-1·d^-1） group， and high-， medium-， and low-dose （26.26， 13.13， 6.565 g·kg^-1·d^-1， respectively） AMD groups， with ten mice per group. Continuous sleep deprivation was administered for 4 weeks via a custom-built sleep deprivation chamber. The cognitive function of mice was assessed via the Morris water maze test. Hematoxylin-eosin （HE） staining was performed to observe morphological alterations in pyramidal neurons. Biochemical assays were carried out to measure the hippocampal and serum levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， catalase （CAT）， and total antioxidant capacity （T-AOC）. Transmission electron microscopy was adopted to examine mitochondrial pathological alterations in the hippocampus. Immunohistochemical assay was conducted to examine the expression of nuclear factor E₂-related factor 2 （Nrf2）， sirtuin 3 （SIRT3）， peroxisome proliferator-activated receptor gamma coactivator-1α （PGC-1α）， mitochondrial transcription factor A （TFAM）， and brain-derived neurotrophic factor （BDNF） in the mouse hippocampus. The protein levels of Nrf2， SIRT3， heme oxygenase-1 （HO-1）， and NAD（P）H：quinone oxidoreductase 1 （NQO1） in mouse hippocampal tissue were determined by Western blot. Immunofluorescence double labeling was employed to determine the protein levels of SIRT3/Nrf2 in mouse hippocampal tissue. ResultsCompared with the control group， the model group exhibited prolonged latency to navigate， increased total swimming distance， reduced number of platform crossings， and shortened time spent in the target quadrant （P<0.05， P<0.01）， disorganized morphology and arrangement of hippocampal neurons， with increased damaged mitochondria， mitochondrial swelling， reduced cristae， and vacuolization， declined levels of SOD， GSH-Px， CAT， and T-AOC and elevated levels of MDA in the hippocampus and serum （P<0.01）， and downregulated protein levels of SIRT3， Nrf2， PGC-1α， TFAM， BDNF， HO-1， and NQO1 （P<0.01）. Compared with the model group， the melatonin group and AMD groups exhibited shortened spatial navigation latency， reduced total swimming distance， increased number of platform crossings， and prolonged activity time in the target quadrant （P<0.05， P<0.01）， reduced neuronal damage and mitochondrial damage in the hippocampal tissue， declined level of MDA and elevated levels of SOD， GSH-Px， CAT， and T-AOC in the hippocampus and serum （P<0.05， P<0.01）， and upregulated protein levels of SIRT3， Nrf2， PGC-1α， TFAM， BDNF， HO-1， and NQO1 in the hippocampus （P<0.05， P<0.01）. ConclusionAMD may improve the learning and memory in aged sleep-deprived mice by mediating mitochondrial oxidative damage through the SIRT3/Nrf2 signaling pathway.

ObjectiveTo investigate whether Anmei Dan （AMD） improves cognitive function and alleviates circadian rhythm disorders in an aged mouse model of sleep deprivation by inhibiting the erythropoietin-producing hepatocellular receptor A4 （EphA4）/Eph receptor-interacting protein A3 （EphrinA3） signaling pathway and its downstream Ras homolog family member A （RhoA）/Rho-associated coiled-coil containing protein kinase （ROCK） signaling， and to analyze its multi-target regulatory characteristics. MethodsAn aged C57 mouse model of sleep deprivation was established by the modified multiple platform water environment method. The mice were randomized into blank， model， rhynchophylline （Rhy， antagonist， 50 mg·kg^-1·d^-1）， and antagonist plus AMD （26.26 g·kg^-1·d^-1） groups. Spontaneous activity was assessed by the open field test， and circadian rhythm activity was monitored. Hematoxylin-eosin （HE） staining and Nissl staining were conducted to observe neuronal morphology in the hippocampal CA1/CA3 regions. Western blot and immunofluorescence assay were employed to determine the protein levels of EphA4， EphrinA3， Neuronal Nuclei Antigen （NEUN）， glial fibrillary acidic protein （GFAP）， phosphorylated （p）-EphA4， RhoA， Rho-associated coiled-coil containing protein kinase 1 （ROCK1）， Rho-associated coiled-coil containing protein kinase 2 （ROCK2）， brain-derived neurotrophic factor （BDNF）， synaptophysin （SYN）， postsynaptic density protein 95 （PSD95）， and growth-associated protein 43 （GAP43）. The mRNA levels of EphA4 and EphrinA3 were determined by real-time PCR. ResultsCompared with the blank group， the model group showed decreased spontaneous activity （P<0.01）， circadian rhythm disruption （P<0.01）， hippocampal neuron disarrangement and reduced Nissl bodies， upregulated protein levels of EphA4， EphrinA3， NEUN， p-EphA4， RhoA， ROCK1， ROCK2， and GFAP （P<0.01）， and downregulated protein levels of NEUN， BDNF， SYN， PSD95， and GAP43 （P<0.01）. Compared with the model group， both the antagonist group and the antagonist plus AMD group showed recovery in the above indicators （P<0.05， P<0.01）， with the combination group outperforming the antagonist group in behavioral， pathological， and molecular expression aspects （P<0.05， P<0.01）. ConclusionAMD can alleviate cognitive and rhythm disorders caused by sleep deprivation by inhibiting the EphA4/EphrinA3 signaling pathway and its downstream RhoA/ROCK signaling， and upregulating the expression of synapse-related proteins such as BDNF. Its combination with an EphA4 antagonist exhibits a synergistic effect， which suggests that the mechanism involves multi-link synergistic intervention.

ObjectiveTo investigate the effects of Anmei Dan （AMD） on the expression of proteins in the C-X3-C motif chemokine ligand 1 （CX3CL1）/C-X3-C chemokine receptor 1 （CX3CR1） signaling pathway， hippocampal inflammation， and learning and memory in an aged sleep-deprived model. MethodsSixty aged C57 mice were randomly assigned into a control group， a model group， a melatonin group （1.3 mg·kg^-1·d^-1）， and high-， medium-， and low-dose AMD groups （26.26， 13.13， 6.565 g·kg^-1·d^-1， respectively）， with 10 mice per group. Continuous sleep deprivation was administered for 4 weeks through a custom-built sleep deprivation chamber. The cognitive function of mice was assessed via the Y-maze test. Histomorphological alterations in pyramidal cells of the hippocampal CA1 and DG regions were examined by hematoxylin-eosin （HE） and Nissl staining. Synaptic morphology in the hippocampus was visualized with Golgi staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of inflammatory cytokines interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the hippocampal tissue as well as the level of interleukin-10 （IL-10） in the prefrontal cortex. Real-time PCR was performed to detect markers of M1-type ［cluster of differentiation （CD） 86 and inducible nitric oxide synthase （iNOS）］ and M2-type ［CD206 and arginase 1 （Arg1）］ macrophages. Western blot was employed to quantify the protein levels of CX3CL1， CX3CR1， phosphorylated nuclear factor κB p65 subunit （p-NF-κB p65）， and phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK） in the hippocampus. Immunofluorescence double staining was performed to co-label CX3CR1 with ionized calcium-binding adapter molecule 1 （Iba-1） for observation of the co-localization of target proteins with microglia. ResultsCompared with the control group， the model group exhibited disrupted daily activity patterns with reduced spontaneous alternation rates （P<0.01）， disorganized morphology and arrangement of hippocampal neurons， accompanied by reduced numbers of Nissl bodies and dendritic spines （P<0.01）， upregulated protein levels of IL-6， IL-10， IL-1β， TNF-α， CX3CL1， CX3CR1， p-NF-κB p65， p-p38 MAPK， and Iba-1 and mRNA levels of CD86 and iNOS （P<0.01）， and downregulated mRNA levels of CD206 and Arg1 （P<0.05，P<0.01）. Compared with the model group， the melatonin group and medium- and high-dose AMD groups showed increased spontaneous alternation rates （P<0.01）， improved neuronal morphology， number， and spine density in the hippocampal CA1 and DG regions （P<0.01）， declined levels of IL-6， IL-10， IL-1β， and TNF-α （P<0.05， P<0.01）， downregulated protein levels of CX3CL1， CX3CR1， p-NF-κB p65， p-p38 MAPK， and Iba-1 and mRNA levels of CD86 and iNOS， and upregulated mRNA levels of CD206 and Arg1 （P<0.01）. ConclusionAMD may improve the learning and memory in aged sleep-deprived mice by mediating neuroinflammation through the CX3CL1/CX3CR1 signaling pathway.

To further investigate the molecular genetic characteristics of enzomatic nasal tumor vi-rus of goats(ENTV-2)in Yunnan Province,this study measured and analyzed the entire genome of ENTV-2 in Yunnan Province.The results showed that the complete genome sequence of the EN-TV-2 YN2023 strain(GenBank accession number:PP682590.1)was successfully obtained.The YN2023 strain has a total length of 7 307 bp and a typical structure of 5'-M5-gag-pro-pol-env-M3-3'.Whole genome sequence homology analysis showed that the nucleotide homology between YN2023 strain and 41 reference strains ranges from 85.3％to 95.5％.The whole genome evolution-ary tree indicates that the YN2023 strain is closely related to the prevalent strains in China,with certain genetic diversity and geographical clustering.The analysis of the gag gene evolutionary tree shows that the gag gene cluster of YN2023 strain is on a branch of the ENTV-2 gag gene,and YN2023 is clustered on the same small branch as enENTV-FJ1 and GDQY2017 strains,with the closest genetic relationship.The env gene evolutionary tree shows that YN2023 is on the same branch as GDQY2017,GDZJ2022,ENTV-2CHN1-6,ENTV-FJ1,and ENTV-FJ3,and is also on the same branch as GDQY2017,indicating a close genetic relationship.Recombination analysis showed that the YN2023 strain underwent a potential recombination event between breakpoint positions 6378-7478 bp,with the Chinese Chongqing strain enENTV-CQ1(OR669623.1)as the primary parent and the Chinese Sichuan strain BH(MT254062.1)as the secondary parent.This study enriches the genomic information of the ENTV-2 strain in Yunnan Province and provides data sup-port for the genetic variation of ENTV-2 in Yunnan Province.

To further investigate the molecular genetic characteristics of enzomatic nasal tumor vi-rus of goats(ENTV-2)in Yunnan Province,this study measured and analyzed the entire genome of ENTV-2 in Yunnan Province.The results showed that the complete genome sequence of the EN-TV-2 YN2023 strain(GenBank accession number:PP682590.1)was successfully obtained.The YN2023 strain has a total length of 7 307 bp and a typical structure of 5'-M5-gag-pro-pol-env-M3-3'.Whole genome sequence homology analysis showed that the nucleotide homology between YN2023 strain and 41 reference strains ranges from 85.3％to 95.5％.The whole genome evolution-ary tree indicates that the YN2023 strain is closely related to the prevalent strains in China,with certain genetic diversity and geographical clustering.The analysis of the gag gene evolutionary tree shows that the gag gene cluster of YN2023 strain is on a branch of the ENTV-2 gag gene,and YN2023 is clustered on the same small branch as enENTV-FJ1 and GDQY2017 strains,with the closest genetic relationship.The env gene evolutionary tree shows that YN2023 is on the same branch as GDQY2017,GDZJ2022,ENTV-2CHN1-6,ENTV-FJ1,and ENTV-FJ3,and is also on the same branch as GDQY2017,indicating a close genetic relationship.Recombination analysis showed that the YN2023 strain underwent a potential recombination event between breakpoint positions 6378-7478 bp,with the Chinese Chongqing strain enENTV-CQ1(OR669623.1)as the primary parent and the Chinese Sichuan strain BH(MT254062.1)as the secondary parent.This study enriches the genomic information of the ENTV-2 strain in Yunnan Province and provides data sup-port for the genetic variation of ENTV-2 in Yunnan Province.

OBJECTIVE To analyze the metabolites of Zhideke granules and speculate its metabolic pathway in rats in vivo. METHODS Male SD rats were randomly divided into blank group and administration group （Zhideke granules， 9.45 g/kg）； they were given ultrapure water or relevant medicine， twice a day， every 6-8 h， for 3 consecutive days. Serum， urine and feces samples of rats were collected， and their metabolites were identified by UPLC-Q-Exactive-MS technique after intragastric administration of Zhideke granules； their metabolic pathways were speculated. RESULTS After intragastric administration of Zhideke granules， 16 prototype components （i.g. irisflorentin， baicalin， chlorogenic acid） and 11 metabolites （i.g. hydration products of kaempferol or luteolin， methylation products of chlorogenic acid， and hydroxylation products of baicalin） were identified in serum， urine and feces of rats. Among them， 8 prototype components and 4 metabolites were identified in serum samples； 10 prototype components and 7 metabolites were identified in urine samples； 8 prototype components and 5 metabolites were identified in the fecal samples. CONCLUSIONS The metabolites of Zhideke granules in rats mainly include baicalin， irisflorentin，chlorogenic acid， and the main metabolic pathways included methylation， hydroxylation， glucuronidation.

OBJECTIVE：To optimize the extraction technology of Zhideke granules. ME THODS：The extraction technology （water extraction ，alcohol extraction ，water extraction and ethanol precipitation ）of Zhideke granules was initially screened by ammonia-induced cough experiment and xylene-induced ear swelling experiment in mice. Based on its preparation route ，the immersion time of medicinal materials containing volatile oil was investigated with water absorption as index firstly. The single factor test was adopted to investigate the amount of water added and the extraction time taking the volatile oil yield as index to optimize the extraction technology of medicinal materials containing volatile oil. Taking the contents of irisflorentin and total flavonoids as indicators ，on the basis of single factor investigation ，orthogonal test was adopted to examine the influence of three factors including the amount of water added ，extraction time and extraction frequency ，so as to optimize the water extraction technology of Zhideke granules and the validation tests were conducted. RESULTS ：The results of pharmacodynamics experiment showed that the cough latency of mice in water extract low-dose and high-dose groups （6.34，12.68 g/kg，by crude drug ）and water-extraction alcohol-precipitation extract high-dose group （12.68 g/kg，by crude drug ）were significantly longer than those inmodel group ，and the number of cough within 2 minutes was significantly reduced （P＜0.05 or P＜0.01）. Compared with model group ， the ear swelling of mice in water extract low-dose and high-dose groups （6.34，12.68 g/kg，by crude drug），ethanol extract high-dose group （12.68 g/kg，by crude drug） and water-extraction alcohol-precipitation extract hig dose group （12.68 g/kg，by crude drug ） were decreased significantly （P＜0.05 or P＜0.01）. The swelling inhibition rates were 42.26％，55.08％，33.49％，51.56％，39.57％ and 44.36％ in low-dose and high-dose groups of water extract ，alcohol extract ， water-extraction and alcohol-precipitation extract respectively ，indicating that the water extract had better antitussive and anti-inflammatory effects. The optimal extraction technology of volatile oil was adding 5-fold water ，soaking for 30 minutes，and extracting for 3 hours. The optimal water extraction technology was adding 12-fold water ，extracting for 3 times after soaked for 50 min，lasting for 1 h each time. Results of 3 times of validation tests showed that average content of irisflorentin in the extract obtained by optimal technology was 76.47 μg/g（RSD＝ 2.15％，n＝3）and the average content of total flavonoids was 92.45 mg/g（RSD＝0.48％，n＝3）. CONCLUSIONS ：The optimal extraction technology of Zhideke granules is stable and feasible.

OBJECTIVE： To study the chemical constituents of ethanol extract from Yao medicine Cissampelopsis spelaeicola. METHODS： The petroleum ether， ethyl acetate and n-butanol fraction from 75％ ethanol extract of C. spelaeicola were isolated and purified by silica gel， SephadexLH-20 gel column and AB-8 macroporous resin column， etc. The structures of the compounds were analyzed and identified by physicochemical properties and spectral data （mass spectrometry， hydrogen spectrum， carbon spectrum）. RESULTS & CONCLUSIONS： Twelve compounds were isolated and identified from 75％ ethanol extract of C. spelaeicola. β-sitosterol（Ⅰ） and Stigmasterol（Ⅱ） were isolated from petroleum ether fraction； p-hydroxybenzoic acid（Ⅲ）， β-daucossterol（Ⅳ）， protocatechuic acid（Ⅴ）， 6β-hydroxyeremophi-7（11）-en-12，8β-olide（Ⅵ）， 10β-hydroxyeremophil-7（11）-en-8，12-olide（Ⅶ）， 10β-hydroxyeremophi-7（11），8（9）-dien-8，12-olide（Ⅷ）， Quercetin（Ⅸ）， Hyperin（Ⅹ） and 4α-hydroxy- eudesman-11-ene（Ⅺ） were isolated from ethyl acetate fraction； quercetin-3-O-robinobioside（Ⅻ） was isolated from n-butanol fraction. Compounds Ⅰ-Ⅻ are isolated from C. spelaeicola for the first time. The study can lay material foundation for activity stady of C. spelaeicola.

Objective To construct Four in One informational teaching model based on cloud platforms, simulation training, nursing micro-lectures and multidimensional evaluation investigate, and to explore the effect of theFour in Oneinformational teaching model applied in Medical Nursing teaching. Methods Two classes students of 2014 were selected by randomized sampling and were divided into experiment group and control group, the experiment group operated the Four in One informational teaching mod, the control group operated regular teaching. After the experiment,using the medical nursing theory test, the skill test, the questionnaire survey to evaluate the teaching effectiveness. Results The theoretical examination score, skill examination score and overall results of experiment group were (78.81±6.44), (82.01±5.22), (80.41±6.32) points, while (71.12±8.12), (76.74±6.3), (75.57±5.51) points of control group, and the difference was statistically significant (t=5.47, 4.86, 3.22, P<0.01 or 0.05). The students' interested in the informational teaching model of the experimental group reached 78.72%(37/47) and learning satisfaction reached 91.48% (43/47). Conclusions The Four in One informational teaching model applied in Medical Nursing teaching can improve the study effect .

Objective To understand the spatial distribution characteristics of wild feces in schistosomiasis endemic areas of Jiangling County,Hubei Province and further explore the source of infection efficiently,so as to provide the evidence for the development of corresponding monitoring and response technology. Methods In 2011,the fresh wild feces were investigated every two months in the selected 15 villages by the severity of historical endemic in Jiangling County. The schistosome miracidi-um hatching method was used to test the schistosome infection of the wild feces. The descriptive analysis and spatial analysis were used for the description of the spatial distribution of the wild feces. Results Totally 701 wild feces samples were collected with the average density of 0.0556/100 m2,and the positive rate of the wild feces was 11.70%(82/701). The results of the re-gression analysis showed a positive spatial correlation between the positive rate of wild feces and the rate of human infection,the area with infected Oncomelania hupensis and the number of fenced cattle,and the corrected R2 of the model was 0.58. Conclu-sion The infection rate of wild feces is positively correlated with the rate of human infection,area with infected O. hupensis and number of fenced cattle in space in Jiangling County,so the prevention and control measures could be conducted according to the spatial distribution of the positive wild feces.