Objective:To evaluate the effect of esketamine on learning and memory function after chronic stress and the signaling pathway of N-methyl-D-aspartate receptor (NMDAR)-calmodulin-dependent protein kinase type 2 (CaMKⅡ)-cAMP-responsive element-binding protein (CREB) in the hippocampus of developing rats.Methods:Sixty clean-grade healthy Sprague-Dawley rats of either sex, aged 7 days, weighing 10-15 g, were divided into 3 groups ( n=20 each) using a random number table method: control+ normal saline group (CN group), chronic stress+ normal saline group (NS group), and chronic stress+ esketamine group (ES group). A chronic stress model was established using a chronic unpredictable stress method. After the end of stress stimulation, esketamine 10 mg/kg was intraperitoneally injected once a day for 7 consecutive days in ES group, and the equal volume of normal saline was given instead in NS group. Y maze test and Morris water maze test were used to assess the learning and memory function after intraperitoneal administration. Venous blood samples were obtained to measure the serum cortisol and reactive oxygen species (ROS) concentrations by enzyme-linked immunosorbent assay. The animals were then sacrificed under anesthesia, the brain was removed and the hippocampal tissue was isolated for examination of the pathological changes in the hippocampal CA1 region and for determination of the ratios of phosphorylated NMDAR (p-NMDAR)/NMDAR, phosphorylated CaMKII (p-CaMKⅡ)/CaMKⅡ, and phosphorylated CREB (p-CREB)/CREB (by Western blot). Results:Compared with CN group, the time spent in the novel arm was significantly shortened, the number of entries into the novel arm was reduced, the escape latency was prolonged, the number of crossing the original platform was reduced, the serum cortisol and ROS concentrations were increased, the p-NMDAR/NMDAR ratio, p-CaMKⅡ/CaMKⅡ ratio and p-CREB/CREB ratio were decreased ( P<0.05), and the pathological changes of neurons were marked in NS group. Compared with NS group, the time spent in the novel arm was significantly prolonged, the number of entries into the novel arm was increased, the escape latency was shortened, the number of crossing the original platform was increased, the serum cortisol and ROS concentrations were decreased, the p-NMDAR/NMDAR ratio, p-CaMKⅡ/CaMKⅡ ratio and p-CREB/CREB ratio were increased ( P<0.05), and the pathological changes of neurons were significantly attenuated in ES group. Conclusions:Esketamine can improve the learning and memory function after chronic stress in developing rats, and the mechanism may be related to reduction of oxidative stress and enhancement of the activity of hippocampal NMDAR-CaMKII-CREB signaling pathway.

Sha （痧） is an acute infectious disease characterised by the appearance of rashes on the skin， caused by exposure to epidemic toxin and pestilent qi. Sha Zhang Yu Heng （《痧胀玉衡》） discussed the treatment principles and methods， and listed collateral bloodletting as one of the main treatments. Through organizing the articles and proved cases， we found that the author believes Sha （痧） is caused by epidemic pathogen， belonging to heat toxin with rapid changes， so timely treatment for qi and blood simultaneously could achieve the effect of transforming qi into defensive qi. Sha Zhang Yu Heng focuses on patient's position during treatmet， the material of the needle， the site of treatment， the quantum of stimulation and the operation of the contraindications and other essentials. According to the depth of the disease location， use traditional Chinese herbal medicine， scraping together to identify the root of the disease. In addition， diet suggestions for the prevention of the recrudescence of disease are also described in detail.

Objective:To evaluate the role of microRNA-149-5p (miR-149-5p) in resveratrol-induced reduction of lipopolysaccharide (LPS)-induced cardiomyocyte injury in rats.Methods:Rat cardiomyocyte cell line H9C2 was cultured and then divided into 5 groups ( n=27 each) using a random number method: control group (C group), LPS group, resveratrol group (RSV group), miR149-5p inhibitor negative control group (LRN group), and miR149-5p inhibitor group (LRI group). A cardiomyocyte injury model was prepared by incubating cells with culture medium containing 10 μg/ml LPS for 24 h. RSV group was incubated with resveratrol (final concentration of 10 μmol/L) for 24 h, followed by incubation with culture medium containing 10 μg/ml LPS for another 24 h. LRN group and LRI group were transfected with miR149-5p inhibitor negative control and miR149-5p inhibitor, respectively, and then the other treatments were similar to those previously described in RSV group. The cell viability was measured by CCK-8 assay, the apoptosis rate by flow cytometry, the concentration of lactate dehydrogenase (LDH) and content of glutathione (GSH) in the supernatant by microplate method, the content of malondialdehyde (MDA) by TBA reaction method, the activity of superoxide dismutase (SOD) by WST-1 method, the level of reactive oxygen species (ROS) by DCFH-DA fluorescent probe, the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant by enzyme-linked immunosorbent assay, and the expression of miR-149-5p by quantitative real-time polymerase chain reaction. Results:Compared with C group, the expression of miR-149-5p was significantly down-regulated, the cell viability was decreased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were increased, and the GSH content and SOD activity were decreased in LPS group ( P<0.05). Compared with LPS group, the expression of miR-149-5p was significantly up-regulated, the cell viability was increased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were decreased, and the GSH content and SOD activity were increased in RSV group ( P<0.05). Compared with RSV group or LRN group, the expression of miR-149-5p was significantly down-regulated, the cell viability was decreased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were increased, and the GSH content and SOD activity were decreased in LRI group ( P<0.05). Conclusions:The mechanism by which resveratrol alleviates LPS-induced cardiomyocyte injury is associated with the up-regulation of miR-149-5p expression and inhibition of cell apoptosis, oxidative stress and inflammatory responses in rats.

Objective:To evaluate the effect of dexmedetomidine on the hippocampal brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrκB) signaling pathway in a rat model of cerebral ischemia-reperfusion (I/R).Methods:Forty-five clean-grade healthy Sprague-Dawley rats, half male and half female, aged 5-6 months, weighing 200-250 g, were divided into 3 groups ( n=15 each) using a random number table method: sham operation group (group S), cerebral I/R group (group I/R), and dexmedetomidine + I/R group (group Dex). A rat model of cerebral I/R injury was established by occluding the middle cerebral artery for 90 min followed by restoring perfusion. In group Dex, dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before ischemia, while the equal volume of normal saline was intraperitoneally injected in S and I/R groups. Neurological deficit scores were evaluated at 12 h of reperfusion. The rats were anesthetized and sacrificed, and the hippocampus was isolated for determination of the percentage of cerebral infarct size (by TTC method), expression of BDNF and TrκB (by Western blot), and expression of BDNF mRNA and TrκB mRNA (by real-time polymerase chain reaction) and for microscopic examination of cell apoptosis (by TUNEL method). Results:Compared with group S, the neurological deficit scores and percentage of cerebral infarct size were significantly increased, the number of apoptotic hippocampal neurons was increased, and the expression of BDNF and TrκB protein and mRNA was down-regulated in I/R and Dex groups ( P<0.05). Compared with group I/R, the neurological deficit scores and percentage of cerebral infarct size were significantly decreased, the number of apoptotic hippocampal neurons was reduced, and the expression of BDNF and TrκB protein and mRNA was up-regulated in group Dex ( P<0.05). Conclusions:The mechanism by which dexmedetomidine alleviates cerebral I/R injury may be related to activating hippocampal BDNF/TrκB signaling pathways in rats.

Objective To investigate the effect of resveratrol(Rsv)on inhibiting miR-149-5p-mediated ferroptosis and impro-ving lipopolysaccharide(LPS)-induced cardiomyocyte(H9C2)injury.Methods Different concentrations of Rsv were used to treat H9C2 cells,followed by LPS stimulation to observe the effect of Rsv on LPS-induced injury in H9C2 cells.The H9C2 cells were divided into different groups,including the Control group,LPS group,LPS+Rsv group,LPS+miRNA-inhibitor-NC group,LPS+miR149-5p-inhibitor group,and LPS+miR149-5p-inhibitor+Rsv group.CCK-8 assay was used to detect cell viability,and a kit was used to measure changes in lactate dehydrogenase(LDH),glutathione(GSH),and malondialdehyde(MDA)release in H9C2 cells.DCFH-DA fluorescent probe was used to determine the level of reactive oxygen species(ROS)in H9C2 cells.Iron ion colorimetry was used to observe changes in Fe2+content in cardiomyocytes,and RT-qPCR was used to detect the expression of miR-149-5p in cells.Results Rsv significantly reduced LPS-induced injury in H9C2 cells.Compared with the LPS group,the Rsv+LPS group showed in-creased cell viability,reduced LDH secretion,increased GSH release,reduced lipid ROS generation,decreased Fe2+content,reduced MDA release,and increased miR-149-5p expression.Compared with the LPS+miR-149-5p-inhibitor group,the LPS+miR-149-5p-inhibitor+Rsv group showed increased cell viability,reduced LDH content,increased GSH,reduced ROS production,re-duced lipid peroxidation and iron accumulation,and increased miR-149-5p expression.Conclusion Rsv may inhibit ferroptosis by upregulating miR-149-5p expression and thus alleviate LPS-induced injury in H9C2 cells.

Objective To investigate the effects of dexmedetomidine on the proliferation and auto-phagy of human colon cancer cells.Methods In experiment 1,human colon cancer cells LoVo and HCT116 were selected and divided into eight groups:LoVo-1 group(group L0-1),LoVo+dexmedetomi-dine 1 nmol/L-1 group(group L1-1),LoVo+dexmedetomidine 10 nmol/L-1 group(group L10-1),LoVo+dexmedetomidine 100 nmol/L-1 group(group L100-1),HCT116-1 group(group H0-1),HCT116+dexmedetomidine 1 nmol/L-1 group(group H1-1),HCT116+dexmedetomidine 10 nmol/L-1 group(group H10-1)and HCT116+dexmedetomidine 100 nmol/L-1 group(group H100-1).The cell proliferation rate was detected by CCK-8 method 24 and 48 hours after drug treatment.The cells were collected 24 hours after drug treatment,and the contents of microtubule-associated protein 1 light chain 3(LC3)-Ⅱ and autophagy associated protein Beclin-1 were detected by Western blot method.In experiment 2,LoVo and HCT116 were selected and divided into four groups:LoVo-2 group(group L0-2),LoVo+dexmedetomidine 10 nmol/L-2 group(group L10-2),HCT116-2 group(group H0-2)and HCT116+dexmedetomidine 10 nmol/L-2 group(group H10-2).Cells were collected 24 hours after drug treatment,LC3 protein expression was observed by immunofluorescence method,and the positive rate of LC3 site was calculated.The autophagosomes were col-lected and observed by transmission electron microscopy 24 hours after drug treatment.Results In experi-ment 1,the cell proliferation rate in groups L10-1 and L100-1 was significantly lower than that in groups L0-1 and L1-1 24 and 48 hours after drug treatment,the protein content of LC3-Ⅱ and Beclin-1 was signifi-cantly higher than that in groups L0-1 and L1-1 24 hours after drug treatment(P＜0.05).The cell prolifer-ation rate in groups H10-1 and H100-1 was significantly lower than that in groups H0-1 and H1-1 24 and 48 hours after drug treatment,and the protein content of LC3-Ⅱ and Beclin-1 was significantly higher than that in groups H0-1 and H1-1 24 hours after drug treatment(P＜0.05).In experiment 2,compared with group L0-2,the positive rate of LC3 site in group L10-2 was significantly increased 24 hours after drug treatment(P＜0.05).Compared with group H0-2,the positive rate of LC3 site in group H10-2 was significantly in-creased 24 hours after drug treatment(P＜0.05).Groups L0-2 and H0-2 have intact cell membranes and clear nuclei.The cell membrane in groups L10-2 and H10-2 was damaged,the arrangement of organelles was disordered,and a large number of autophagosomes and autophagosomes were visible.Conclusion Dexmedetomidine may inhibit the proliferation of colon cancer cells by inducing autophagy.

Objective To study the requirement of exogenous phosphorus for 2-year-old and 3-year-old ginseng seedings.Methods Two-and three-year-old ginsengs were used as experimental materials.Calcium superphosphate and phosphorus-free Hoagland solution were used as fertilizer,and the concentrations of P2O5 were set to be 0 mmol·L-1,0.5 mmol·L-1,1.5 mmol·L-1,3 mmol·L-1,6 mmol·L-1,8 mmol·L-1,respectively.The effects of different concentrations of phosphorus fertilizer on agronomic indexes,photosynthetic characteristics and accumulation of ginsenosides Rg1,Rb1 and Re were studied.Results The plant height,stem diameter,root weight,leaf area,relative content of chlorophyll,net photosynthetic rate and total saponins content of different year-old panax ginseng were significantly increased by applying phosphorus at 1.5-3.0 mmol·L-1.Among them,compared with the phosphorus-free group,the root weight of second-year ginseng was increased by 16％and the saponin content was increased by 24％;the root weight of third-year ginseng was increased by 89％and the saponin content was increased by 132％.The appropriate application rate of phosphorus fertilizer(phosphorus pentoxide)during the growth of second and third year ginseng was 26.6 mg and 53.3 mg of plant,respectively.Conclusion External application of suitable concentration of phosphorus fertilizer can enhance the external morphological characteristics of ginseng,improve photosynthetic physiological properties and increase the content of active ingredients.

Objective:To evaluate the effect of intravenous infusion of lidocaine on pulmonary gas exchange function during acute lung injury in septic rats.Methods:Thirty clean-grade healthy male Sprague-Dawley rats, aged 2-3 months, weighing 220-280 g, were divided into 3 groups ( n=10 each) using a random number table method: control group (C group), sepsis group (S group) and lidocaine group (L group). Model of acute lung injury in septic rats was prepared by intraperitoneal injection of LPS 10 mg/kg in S and L groups, the equal volume of normal saline was injected in group C, lidocaine was injected at a loading dose of 10 mg/kg via the tail vein at 1 min after LPS injection and then continuously infused for 3 h at a rate of 10 mg·kg -1·h -1, and the equal volume of normal saline was given instead in C and S groups. Rats were sacrificed at 24 h after LPS injection, blood samples from the abdominal aorta were collected for blood gas analysis, and the oxygenation index (OI), alveolar-arterial oxygen difference (A-aDO 2) and respiratory index (RI) were calculated. Then the chest was immediately opened, and the left lung tissues were then immediately removed to determine the levels of tumor necrosis factor-α (TNF-α), heme oxygenase-1 (HO-1) and reactive oxygen species (ROS) (by enzyme-linked immunosorbent assay). The right upper lung tissues were removed for microscopic examination of the alveolar structure and pulmonary edema with a light microscope. The right lower lung tissues were also removed to observe the vascular endothelial structure with a transmission electron microscope. Results:Compared with group C, the levels of A-aDO 2, RI, TNF-α, HO-1 and ROS were significantly increased, the PaO 2 and OI were decreased ( P<0.05), and no significant change was found in PaCO 2 in group S and group L ( P>0.05). Compared with group S, the PaO 2, OI and HO-1 were significantly increased, the levels of A-aDO 2, RI, TNF-α and ROS were decreased ( P<0.05), and no significant change was found in PaCO 2 levels in group L ( P>0.05). The pathological damages of the lung tissues were significantly attenuated in group L when compared with group S. Conclusions:Intravenous infusion of lidocaine can improve pulmonary gas exchange function during acute lung injury in septic rats.

Objective:To evaluate the relationship between microRNA-27a (miR-27a) and silent information regulator 1 (SIRT1) during myocardial ischemia-reperfusion (I/R) in rats.Methods:Fifty clean-grade healthy male Sprague-Dawley rats, aged 2-3 months, weighing 220-280 g, were divided into 5 groups ( n=10 each) by the random number table method: sham operation group (Sham group), myocardial I/R group (I/R group), AAV9-miR-27a overexpression + myocardial I/R group (AAV+ I/R group), miR-27a antagomir + myocardial I/R group (AG+ I/R group) and AAV9-miR-27a negative control+ myocardial I/R group (NC+ I/R group). The myocardial I/R injury model was prepared by ligating the anterior descending branch of the left coronary artery for 30 min followed by 120 min reperfusion. At day 14 before ischemia, AAV9-miRNA-27a adeno-associated virus 2×10 11 v. g was injected via the tail vein in AAV+ I/R group, and AAV9-miR-27a NC 2×10 11 v. g was injected via the tail vein in NC+ I/R group. miR-27a antagomir 10 mg/kg was injected via the tail vein once a day at 3 days before ischemia in AG+ I/R group. At the end of 120 min of reperfusion, serum cardiac troponin T(cTnT), creatine kinase isoenzymes (CK-MB) and lactic dehydrogenase (LDH) concentrations and contents of glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MDA) in myocardial tissues were determined by enzyme-linked immunosorbent assay, the percentage of myocardial infarct volume by TTC staining, the expression of miR-27a in myocardial tissues by quantitative real-time polymerase chain reaction, and the expression of SIRT1 in myocardial tissues by Western blot. Results:Compared with Sham group, the percentage of myocardial infarct volume and serum concentrations of cTnT, CK-MB and LDH were significantly increased, the contents of GSH and SOD in myocardial tissues were decreased, MDA contents were increased, miR-27a expression was up-regulated, and SIRT1 expression was down-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of myocardial infarct volume and serum concentrations of cTnT, CK-MB and LDH were significantly increased, the contents of GSH and SOD in myocardial tissues were decreased, MDA contents were increased, miR-27a expression was up-regulated, and SIRT1 expression was down-regulated in AAV+ I/R, and the percentage of myocardial infarct volume and serum concentrations of cTnT, CK-MB and LDH were significantly decreased, the contents of GSH and SOD in myocardial tissues were increased, MDA contents were decreased, miR-27a expression was down-regulated, and SIRT1 expression was up-regulated in AG+ I/R group ( P<0.05), and no significant change was found in the parameters mentioned above in NC+ I/R group ( P>0.05). Compared with AAV+ I/R group, the percentage of myocardial infarct volume and serum concentrations of cTnT, CK-MB and LDH were significantly decreased, the contents of GSH and SOD in myocardial tissues were increased, MDA contents were decreased, miR-27a expression was down-regulated, and SIRT1 expression was up-regulated in AG+ I/R group ( P<0.05). Conclusions:miR-27a is involved in the pathophysiological mechanism underlying myocardial I/R injury probably through inhibition of SIRT1 expression in rats.

Objective:To evaluate the effect of esketamine on long-term cognitive dysfunction induced by propofol anesthesia in the developing rats and the role of phosphatidylinositol-3-kinase (PI3K)/serine-threonine protein kinase (Akt) signaling pathway.Methods:Forty-eight clean-grade healthy Sprague-Dawley rats of either sex, aged 7 days, weighing 10-15 g, were divided into 4 groups ( n=12 each) using a random number table method: fat emulsion group (C group), propofol group (P group), esketamine + propofol group (EP group), and PI3K inhibitor LY294002 + esketamine + propofol group (LYEP group). Medium/long-chain fat emulsion injection 100 mg/kg was intraperitoneally injected in C group. Propofol was intraperitoneally injected at a dose of 50 mg/kg, followed by an additional dose of 50 mg/kg after the righting reflex was restored (40-60 min later) in P group. In group EP, esketamine 10 mg/kg was intraperitoneally injected, followed by propofol administration using the same method as previously described in P group. In LYEP group, LY294002 25 μg was injected via the lateral ventricle, 30 min later ketamine 10 mg/kg was intraperitoneally injected, and then propofol was given using the same method as previously described in P group. Six rats in each group were randomly sacrificed at 2 h after emergence for microscopic examination of pathological changes of hippocampal neurons and for determination of Akt, phosphorylated Akt (p-Akt), Bax, and cleaved caspase-3 in the hippocampal tissues (using Western blot). The remaining 6 rats in each group were subjected to Y-maze test to evaluate their learning and memory abilities at 30 days after birth. The p-Akt/Akt ratio was calculated. Results:Compared with C group, the p-Akt/Akt ratio in the hippocampal tissues was significantly decreased, the expression of Bax and cleaved caspase-3 was up-regulated, the number of training sessions required for learning was increased, the correct response rate was decreased ( P<0.05), and the pathological damage to neurons in hippocampal CA1 region was found in P, EP and LYEP groups. Compared with P group, the p-Akt/Akt ratio in the hippocampal tissues was significantly increased, the expression of Bax and cleaved caspase-3 was down-regulated, the number of training sessions required for learning was decreased, the correct response rate was increased ( P<0.05), and the pathological damage to neurons in hippocampal CA1 region was significantly attenuated in EP and LYEP groups. Compared with EP group, the p-Akt/Akt ratio in the hippocampal tissue was significantly decreased, and the expression of Bax and cleaved caspase-3 was up-regulated, the number of training sessions required for learning was increased, the correct response rate was decreased ( P<0.05), and the pathological damage to neurons in hippocampal CA1 region was aggravated in LYEP group. Conclusions:Esketamine can alleviate long-term cognitive impairment caused by propofol anesthesia in the developing rats, and the mechanism may be related to activation of the PI3K/Akt signaling pathway and inhibition of apoptosis in neurons.