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Objective:This study aims to explore the feasibility and practicality of ChatGPT in scientific research statistics through examples, in order to better understand and evaluate the application value of ChatGPT in scientific research statistics.Methods:Literature research was used to illustrate the principles of ChatGPT artificial intelligence model, combined with practical examples and tests, to assess the reliability of its results and analyze the advantages and limitations of ChatGPT in scientific research statistics.Results:ChatGPT can be applied to various scenarios in scientific research statistics, such as literature retrieval and organization, experimental design, statistical analysis programming, and statistical graphing. It can provide efficient scientific research statistical analysis capabilities, but it also has limitations.Conclusions:Although there are limitations, ChatGPT has a promising future in scientific research statistics, and it will be applied widely and deeply to greatly facilitate scientific research.
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Objective:To evaluate the efficacy of esketamine combined with fascia iliaca compartment-subarachnoid block in optimizing anesthesia in elderly patients undergoing hip fracture surgery.Methods:Sixty-two American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ elderly patients of either sex, aged 60-85 yr, with body mass index of 18.5-30.0 kg/m 2, were divided into 2 groups ( n=31 each) using a random number table method: fascia iliaca compartment-subarachnoid block group (FS group) and esketamine combined with fascia iliaca compartment-subarachnoid block group (ES group). In FS group, patients underwent ultrasound-guided fascia iliaca compartment block at 30 min before the operation of subarachnoid anesthesia on the surgical side. In ES group, esketamine 0.25 mg/kg was intravenously administered at 5 min before skin incision based on the fascia iliaca compartment-subarachnoid block. Patient-controlled intravenous analgesia was used for postoperative analgesia, and tramadol 1 mg/kg was intravenously given for rescue analgesia when numerical rating scale score > 4. The pressing times of patient-controlled analgesic pump, the number of rescue analgesia and consumption of tramadol were recorded within 48 h after operation. The occurrence of postoperative adverse reactions (respiratory depression, nausea and vomiting, dizziness, drowsiness, pruritus, illusion, nightmares) was recorded. Results:Compared with FS group, the consumption of postoperative tramadol was significantly decreased, and the pressing times of patient-controlled analgesic pump and the number of rescue analgesia were reduced in ES group ( P<0.05). There were no significant differences in the incidence of postoperative adverse reactions between the two groups ( P>0.05). Conclusions:Combination of esketamine with fascia iliaca compartment-subarachnoid block for hip fracture surgery can raise postoperative analgesia and optimize clinical management strategies in elderly patients.
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Objective:To evaluate the role of transient receptor potential vanillic acid 4 (TRPV4) in dexmedetomidine-induced improvement in cognitive function in mice with mechanical ventilator-caused brain injury.Methods:Ninety clean-grade healthy male C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were divided into 5 groups ( n=18 each) using a random number table method: control group (group C), mechanical ventilation group (group V), HC-067047 group (group H), dexmedetomidine group (group D), and dexmedetomidine+ GSK1016790A group (group DG). In group C, the animals breathed air spontaneously for 6 h without mechanical ventilation. In group V, the animals were mechanically ventilated for 6 h. In group H, TRPV4 blocker HC-067047 10 mmol was injected into the cerebral ventricle at 3 and 6 h of mechanical ventilation. In D and DG groups, dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation. In group DG, TRPV4 agonist GSK1016790A 5 μmol was injected into the cerebral ventricle at 60 min before mechanical ventilation. Morris water maze test was performed on 6 mice in each group at 1 day before mechanical ventilation and 3 and 7 days after mechanical ventilation. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the brain tissue was taken for determination of the neuronal apoptosis in hippocampal CA1 area by TUNEL method, and the apoptosis index was calculated. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the hippocampal tissues were taken for determination of the expression of TRPV4, serine-threonine protein kinase (Akt), phosphorylated Akt (p-Akt), Bcl-2, Bax and caspase-3 by Western blot. Results:Compared with group C, the escape latency was significantly prolonged and the number of crossing the original platform was reduced at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was up-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group V and group DG ( P<0.05). Compared with group V, the escape latency was significantly shortened and the number of crossing the original platform was increased at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was down-regulated, the expression of p-Akt was up-regulated, the ratio of Bcl-2/Bax was increased, and the apoptosis index of neurons was decreased in group D and group H ( P<0.05). Compared with group D, the escape latency was significantly prolonged at 3 and 7 days after mechanical ventilation, the number of crossing the original platform was reduced, the expression of TRPV4 and caspase-3 was up-regulated, the expression of p-Akt was down-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group DG ( P<0.05). Conclusions:TRPV 4 is involved in dexmedetomidine-induced improvement in cognitive function, which is related to up-regulation of p-Akt expression and inhibition of apoptosis in hippocampal neurons in mice with mechanical ventilation-caused brain injury.
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Objective:To evaluate the role of transient receptor potential vanilloid receptor 1 (TRPV1)/nuclear factor-κB (NF-κB) signaling pathway in dexmedetomidine-induced alleviation of ventilator-induced lung injury (VILI) in rats.Methods:One hundred clean-grade healthy male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 5 groups ( n=20 each) using a random number table method: control group (group C), VILI group (group V), AMG9810 group (group A), dexmedetomidine group (group D), and dexmedetomidine + RTX group (group DR). VILI model was prepared by mechanical ventilation with a tidal volume of 40 ml/kg for 4 h. In group A, TRPV1 inhibitor AMG9810 30 mg/kg was intraperitoneally injected at 1 h before mechanical ventilation.Dexmedetomidine 5.0 μg/kg was intravenously infused at 20 min before mechanical ventilation, and dexmedetomidine was intravenously infused at the rate of 5.0 μ g·kg -1·h -1 during ventilation in group D and group DR.In group DR, RTX 70 μ g/kg was intraperitoneally injected for 3 consecutive days before mechanical ventilation.At 4 h of mechanical ventilation, the concentrations of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 in bronchoalveolar lavage fluid (BALF) were detected, oxygenation index (OI) and wet/dry lung weight (W/D) ratio were measured, the histopathological changes of lung tissues were observed, and lung injury was assessed and scored.The expression of TRPV1 and NF-κB in lung tissues was detected by Western blot, and real-time polymerase chain reaction was used to detect the expression of TRPV1 and NF-κB mRNA. Results:Compared with group C, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly increased, OI was decreased, the W/D ratio and lung injury scores were increased, and the expression of TRPV1 and NF-κB protein and mRNA was up-regulated in group V ( P<0.05). Compared with group V, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly decreased, OI was increased, the W/D ratio and lung injury scores were decreased, and the expression of TRPV1 and NF-κB protein and mRNA was down-regulated in A, D and DR groups ( P<0.05). Compared with group D, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly increased, OI was decreased, the W/D ratio and lung injury scores were increased, and the expression of TRPV1 and NF-κB protein and mRNA was up-regulated in group DR ( P<0.05). Conclusions:The mechanism by which dexmedetomidine alleviates VILI is partially related to inhibition of the activation of TRPV1/NF-κB signaling pathway and inhibition of the inflammatory responses in lung tissues of rats.
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Objective:To evaluate the effects of dexmedetomidine on alveolar epithelial barrier function in rats with ventilator-induced lung injury (VILI), and the role of protein kinase C (PKC).Methods:One hundred clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 5 groups ( n=20 each) using a random number table method: control group (group C), VILI group (group V), PKC inhibitor group (group B), dexmedetomidine group (group D), and dexmedetomidine plus PKC agonist group (DP group). The VILI model was developed by mechanical ventilation with a tidal volume of 40 ml/kg for 4 h in anesthetized animals.Group C breathed air autonomously for 4 h without mechanical ventilation.Group V was mechanically ventilated for 4 h. In group B, bisindolvlmaleimide I 0.12 mg/kg was injected intramuscularly 1 h before mechanical ventilation.In D and DP groups, dxmedetomidine 5.0 μg/kg was injected intravenously at 20 min before mechanical ventilation, and dexmedetomidine was intravenously infused at the rate of 5.0 μg·kg -1·h -1 during mechanical ventilation.In group DP, PKC agonist phorbol-12-myristic acid-13-acetate 15 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation.At 4 h of mechanical ventilation, oxygenation index (OI), lung permeability index (LPI) and wet/dry lung weight (W/D) ratio were measured, the pathological changes of lung tissues were observed, and lung injury was assessed and scored.The expression of PKC, occludin and ZO-1 protein was detected by Western blot, and the expression of PKC mRNA, occludin mRNA and ZO-1 mRNA was determined by real-time polymerase chain reaction. Results:Compared with group C, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in V and DP groups ( P<0.05), and no significant change was found in the parameters mentioned above in B and D groups ( P>0.05). Compared with group V, OI was significantly increased, LPI, W/D ratio and lung injury score were decreased, the expression of PKC protein and mRNA was down-regulated, and the expression of occludin and ZO-1 protein and mRNA was up-regulated in B, D and DP groups ( P<0.05). Compared with group D, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in group DP ( P<0.05). Conclusions:Dexmedetomidine can reduce the damage to alveolar epithelial barrier function in rats with VILI, and the mechanism is related to inhibition of PKC activation and up-regulation of the expression of occludin and ZO-1.
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Objective:To investigate the effect of baicalin on cognitive function of mice with brain injury induced by mechanical ventilation and its mechanism.Methods:Seventy two C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were randomly divided into control group (group C), mechanical ventilation group (group V), baicalin group (group B), baicalin+ Akt inhibitor MK-2206 group (group BM) according to random number table method, with 18 in each group.Mice in group C did not have mechanical ventilation and breathed air independently for 6 hours.Mice in group V received mechanical ventilation for 6 hours.Mice in group B and group BM were intraperitoneally injected with baicalin 100 mg/kg 30 minutes before mechanical ventilation, and mice in group BM were injected intraventricular with Akt inhibitor MK-2206 300 μg/kg 60 minutes before mechanical ventilation.Six mice in each group were randomly selected to test their learning and memory abilities by Morris water maze test 1st day before mechanical ventilation and 3rd day and 7th day after mechanical ventilation.One day after mechanical ventilation, six mice in each group were killed, and the brain tissue was taken.TUNEL method was used to detect the neuronal apoptosis in hippocampal CA1 area, and the apoptosis index was calculated.One day after mechanical ventilation, six mice in each group were killed, and the hippocampus was taken, Western blot was used to detect the protein expressions of caspase-3, caspase-9, Akt, p-Akt, GSK-3β and p-GSK-3β.SPSS 22.0 software was used for statistical analysis of data, repeated measure ANOVA and one-way ANOVA were used for comparison between multiple groups.LSD- t test was used for further pairwise comparison. Results:The results of water maze test showed that the time and group interaction of the four groups were not significant ( F=1.14, P>0.05), the main effect of time and group were both significant ( F=47.36, 59.65, both P<0.05). At 3rd day and 7th day after mechanical ventilation, the escape latencies of mice in group V were higher than those in group C (both P<0.05), and the numbers of platform crossing were lower than those in group C (both P<0.05). And 3 days and 7 days after mechanical ventilation, the escape latencies of mice in group B were lower than those in group V (both P<0.05) and the numbers of platform crossing were higher than those in group V (both P<0.05). The escape latenies of mice in BM group on the 3rd and 7th day were higher than those in group B (both P<0.05), and the numbers of platform crossing were lower than those in group B on the 3rd day and 7th day after mechanical ventilation(both P<0.05). TUNEL and Western blot results showed that apoptosis index of hippocampal neurons and expression levels of apoptosis-related proteins caspase-3 and caspase-9 were significant different in the four groups ( F=51.42, 41.21, 40.19, all P<0.05). The apoptosis index of hippocampal neurons ((40.6±3.9)%), the expression levels of caspase-3 (4.93±0.92) and caspase-9 (4.81±0.88) in the hippocampus of mice in group V were higher than those in group C ((13.7±1.4)%, (1.87±0.27), (1.71±0.25), all P<0.05), the apoptosis index of hippocampal neurons ((15.6±1.6)%), the expression levels of caspase-3 (1.95±0.30) and caspase-9 (1.76±0.28) in group B were lower than those in group V ((40.6±3.9)%, (4.93±0.92), (4.81±0.88), all P<0.05), the apoptosis index of hippocampal neurons ((27.8±2.7)%), the expression levels of caspase-3 (3.58±0.61) and caspase-9 (3.49±0.57) in BM group were higher than those in group B ((15.6±1.6)%, (1.95±0.30), (1.76±0.28), all P<0.05). Expression level of p-Akt, p-GSK-3β in hippocampal tissues of the four group of mice were significantly different ( F=37.54, 43.23, both P<0.05). The expression level of p-Akt (0.51±0.06) and p-GSK-3β (0.47±0.05) of hippocampal tissues of mice in group V were lower than those of group C ((1.07±0.10), (1.11±0.12), both P<0.05), the expression level of p-Akt (0.99±0.10) and p-GSK-3β (1.08±0.09) of hippocampal tissues of mice in group B were higher than those of group V (both P<0.05), the expression level of p-Akt (0.83±0.08) and p-GSK-3β (0.81±0.07) of hippocampal tissues of mice in group BM were lower than those in group B (both P<0.05). Conclusion:Baicalin can improve the cognitive function of mice with brain injury induced by mechanical ventilation, which is related with activation of Akt/GSK-3β signaling pathway and inhibition of hippocampal neuron apoptosis.
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Objective:To evaluate the relationship between the mechanism underlying methylprednisolone-induced alleviation of ventilator-induced lung injury (VILI) and p38 mitogen-activated protein kinase (p38 MAPK)/nucleotide binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) pathway in lung tissues of rats.Methods:Sixty clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), mechanical ventilation group (group V), and methylprednisolone group (group M). Group C breathed air spontaneously for 4 h without mechanical ventilation.Group V was mechanically ventilated (RR 40 times/min, V T 40 ml/kg, I∶E 1∶1, PEEP 0, FiO 2 21%) for 4 h. Group M received intravenous methylprednisolone 10 mg/kg at 20 min before mechanical ventilation.At 4 h of mechanical ventilation, broncho-alveolar lavage fluid (BALF) was collected to measure the concentrations of interleukin-1beta (IL-1β), IL-18, and tumor necrosis factor-alpha (TNF-α) and wet/dry lung weight ratio (W/D ratio), and lung tissues were obtained for microscopic examination of the histopathological changes and for detection of the expression of p38MAPK, phosphorylated p38MAPK (p-p38MAPK), NLRP3, apoptosis-related speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific protease-1 (caspase-1) (using Western blot). Results:Compared with group C, the W/D ratio of lung tissues and concentrations IL-1β, IL-18 and TNF-α in BALF were significantly increased, and the expression of p-p38MAPK, NLRP3, ASC and caspase-1 was up-regulated in group V ( P<0.05), and no significant change was found in group M ( P>0.05). Compared with group V, the W/D ratio of lung tissues and concentrations of IL-1β, IL-18 and TNF-α in BALF were significantly decreased, and the expression of p-p38MAPK, NLRP3, ASC and caspase-1 was down-regulated in group M ( P<0.05). Conclusion:The mechanism by which methylprednisolone alleviates VILI may be related to inhibition of p38MAPK/NLRP3 pathway activity and reduction of inflammatory responses in lung tissues of rats.
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Objective:To evaluate the effect of carbon monoxide-releasing molecule-3 (CORM-3) on blood transfusion-related acute lung injury in rats with traumatic brain injury (TBI).Methods:Seventy-two clean-grade healthy adult male Sprague-Dawley rats, weighing 300-350 g, were divided into 4 groups ( n=18 each) using the random number table method: sham operation group (group S), TBI group (T group), TBI plus 10 ml/kg plasma transfusion group (TP group), and TBI plus 10 ml/kg plasma transfusion plus CORM-3 group (TPC group). TBI was induced by dropping a 20-g weight from 20 cm height falling freely in anesthetized rats.Plasma 10 ml/kg was infused via the femoral vein after TBI in TP and TPC groups.The rats were sacrificed at 24 h after plasma transfusion, and lung tissues were obtained for determination of wet/dry weight (W/D) ratio, cell apoptosis, and expression of caspase-3, Bid, Bim and Puma (by Western blot). The lung injury score was calculated using the results of HE staining.Lung ultrasonography was performed for assessment of sonographic score, and the apoptosis rate was calculated by the TUNEL staining method. Results:Compared with S group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in the other three groups ( P<0.05). Compared with T group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in TP group ( P<0.05). Compared with TP group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly decreased, and the expression of activated caspase-3, Bid, Bim and Puma was down-regulated in TPC group ( P<0.05). Conclusion:CORM-3 can reduce acute lung injury related to blood transfusion in rats with TBI, and the mechanism may be related to inhibiting cell apoptosis in lung tissues.
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Objective:To evaluate the effect of dexmedetomidine on the extracellular signal-regulated kinase(ERK)/sodium-potassium ATPase(Na + -K + -ATPase)signaing pathway in lung tissues of rats with mechanical ventilation-induced lung injury (VILI). Methods:Forty-eighty clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 4 groups ( n=12 each) using a random number table method: control group (group C), VILI (alpha2-adrenergic receptor antagonist) group (group V), dexmedetomidine group (group D), and dexmedetomidine plus yohimbine group (group DY). Group C underwent no mechanical ventilation and breathed air spontaneously for 4 h. Mechanical ventilation (respiratory rate 40 breaths/min, tidal volume 40 ml/kg, inspiratory/expiratory ratio 1∶1, PEEP 0, fraction of inspired oxygen 21%) lasted 4 h in group V. Dexmedetomidine was infused intravenously in a dose of 5.0 μg/kg at 20 min before ventilation followed by an infusion of 5.0 μg·kg -1· h -1 throughout ventilation in group D. In group DY, yohimbine 0.1 mg/kg was injected intravenously at 10 min before dexmedetomidine, and the other treatments were similar to these previously described in group D. Blood samples and lung tissues were taken at 4 h of mechanical ventilation to determine the wet/dry weight ratio (W/D ratio), lung permeability index (LPI), alveolar fluid clearance rate (AFC), and expression of extracellular signal-regulated kinase (ERK), phosphorylated extracellular signal-regulated kinase (p-ERK), and Na + -K + -ATPase in lung tissues (by Western blot) and to observe pathological changes of lung tissues. Results:Compared with group C, LPI and W/D ratio were significantly increased, AFC was decreased, p-ERK expression was up-regulated, and Na + -K + -ATPase expression was down-regulated in group V and group DY ( P<0.05), and no significant change was found in the incidence of the parameters mentioned above in group D ( P>0.05). Compared with group V, LPI and W/D ratio were significantly decreased, AFC was increased, p-ERK expression was down-regulated, Na + -K + -ATPase expression was up-regulated ( P<0.05), and the pathological changes of lung tissues were significantly attenuated in group D, and no significant change was found in the incidence of the parameters mentioned above in group DY ( P>0.05). Compared with group D, LPI and W/D ratio were significantly increased, AFC was decreased, p-ERK expression was up-regulated, Na + -K + -ATPase expression was down-regulated ( P<0.05), and the pathological changes of lung tissues were accentuated in group DY. Conclusion:The mechanism by which dexmedetomidine alleviates VILI may be related to activating alpha2-adrenergic receptors and inhibiting ERK/Na + -K + -ATPase signaling pathway in rats.
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Objective:To evaluate the effect of VX-765 on cognitive function in acute rapid eye movement (REM) sleep-deprived juvenile rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 3-4 weeks, weighing 52-101 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), acute REM group (group AREM) and VX-765 group (group V). Sleep deprivation model was established by modified multi-platform water environment method.In group V, VX-765 solution 10 mg/kg was intravenously injected via the tail vein at 9: 00 a. m.every day for 4 consecutive days.The equal volume of normal saline was given instead in C and AREM groups.Morris water maze and novel object recognition tests were performed for 4 consecutive days during sleep deprivation.The rats were then sacrificed after the end of Morris water maze and novel object recognition tests on 5th day, and hippocampi were removed for determination of the expression of interleukin-1beta (IL-1β) and IL-18 by Western blot. Results:Compared with group C, the latency of novel object recognition was significantly prolonged, the percentage of novel object exploration was shortened, and the number of head exploration was decreased, the percentage of novel object exploration and discrimination index were decreased, the number of crossing the original platform in Morris water maze test was reduced, the time of staying at the target quadrant was shortened, and the expression of IL-1β and IL-18 was up-regulated in AREM and V groups ( P<0.05). Compared with group AREME, the latency of novel object recognition was significantly shortened, the percentage of novel object exploration was prolonged, and the number of head exploration was increased, the percentage of novel object exploration and discrimination index were increased, the number of crossing the original platform in Morris water maze test was increased, the time of staying at the target quadrant was prolonged, and the expression of IL-1β and IL-18 was down-regulated ( P<0.05). Conclusion:VX-765 can improve the cognitive function in acute REM sleep-deprived juvenile rats, which is related to inhibiting hippocampal inflammatory responses.
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Objective To evaluate the role of hippocampal extracellular signal-regulated kinase 1/2 (ERK1/2) in exogenous carbon monoxide (CO)-induced improvement of cognitive function in a rat model of hemorrhage shock and resuscitation.Methods Ninety clean-grade healthy male Sprague-Dawley rats,aged 9-10 weeks,weighing 350-400 g,were divided into 5 groups (n=18 each) using a random number table method:sham operation group (S group),hemorrhage shock and resuscitation group (H group),CO group,PD98059-CO group (PCO group) and PD98059 group (P group).Hemorrhagic shock was induced by withdrawing blood from the femoral vein until mean arterial pressure was reduced to 25-35 mmHg which was maintained for 60 min and resuscitated by infusing the blood withdrawn over 15 min until the initial blood pressure was achieved,and normal saline was infused when needed.Rats were exposed to air mixture containing 1％ CO for 3 h in a glass box after the end of resuscitation in group CO.ERK1/2 inhibitor PD98059 (30 μmol/L) 30 μl was injected into the cerebral ventricle at 30 min before hemorrhage in PCO and PC groups.Right femoral artery and vein were only cannulated,and normal saline was injected into the cerebral ventricle in group S.Rats were sacrificed at 3 h after the end of resuscitation,brains were removed and hippocampi were isolated for determination of CO content (by gas chromatograph assay).Cognitive function was assessed by Morris water maze test at 15 days after the end of resuscitation,rats were then sacrificed and hippocampi were isolated for determination of cell apoptosis in hippocampal CA1 region by TUNEL and cleaved caspase-3 immunofluorescence,and the apoptosis rate was calculated.Rats were sacrificed at 6 h after the end of resuscitation,and hippocampi were isolated to detect the expression of phosphorylated ERK1/2 (p-ERK1/2),Bcl-2 and Bax by Western blot.Results Compared with group S,the escape latency was significantly prolonged in H,PCO and P groups,and the hippocampal CO content and apoptosis rate were increased,the expression of cleaved caspase-3 and ERK1/2 was up-regulated,and the Bcl-2/Bax ratio was decreased in H,CO,PCO and P groups (P＜0.05).Compared with group H,the escape latency was significantly shortened,the hippocampal CO content was increased,and the apoptosis rate was decreased,the cleaved caspase-3 expression was down-regulated,p-ERK1/2 expression was upregulated,and Bcl-2/Bax ratio was increased in group CO,and the escape latency was significantly prolonged,the hippocampal CO content was decreased,the apoptosis rate was increased,the cleaved caspase-3 expression was up-regulated,p-ERK1/2 expression was down-regulated,and Bcl-2/Bax ratio was decreased in group P (P＜0.05).Compared with group CO,the escape latency was significantly prolonged,the apoptosis rate was increased,the cleaved caspase-3 expression was up-regulated,p-ERK1/2 expression was down-regulated,and Bcl-2/Bax ratio was decreased in group PCO (P＜0.05).Conclusion The mechanism by which exogenous CO improves cognitive function is related to raising the phosphorylation of ERK1/2 in hippocampal neurons and inhibiting neuronal apoptosis in a rat model of hemorrhage shock and resuscitation.
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Objective To evaluate the effect of carbon monoxide (CO) postconditioning on pyroptosis induced by oxygen-glucose deprivation and restoration (OGD/R) in rat hippocampai neurons and the relationship with mitochondrial permeability transition pore (mPTP)/reactive oxygen species (ROS) signaling pathway.Methods Primary hippocampal neurons were cultured in vitro,seed in 6-well or 96-well plates,and divided into 5 groups (n =24 each) using a random number table method:control group (C group),OGD/R group,CO postconditioning group (CO group),specific mPTP opener atractyloside plus CO postconditioning group (ACO group),and specific ROS inducer antimycin A plus CO postconditioning group (KCO group).Neurons were subjected to O2-glucose deprivation (OGD) for 16 h followed by restoration of O2-glucose supply for 24 h to establish the model of OGD/R injury.In group CO,neurons were exposed to 2％ CO-5％ CO2 for 3 h at 37 ℃ starting from the end of OGD,followed by normal culture for 21 h.In ACO and KCO groups,atractyloside 20 μmol/L and antimycin A 50 μmol/L were added at the end of OGD,respectively,and the other treatments were similar to those previously described in group CO.Neuronal pyroptosis rate was determined using double immunofluorescent staining cleaved caspase-1-AlexaFluor 568/DAPI after the end of treatments in each group.The neuronal survival rate was determined by MTT,opening of mPTP by Calcein-AM fluorescence,ROS content by DCFH-DA,and expression of interleukin1beta (IL-1β) and IL-18 by Western blot.Results Compared with C group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in the other groups (P＜0.05).Compared with OGD/R group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly decreased,the neuronal survival rate was increased,and the expression of IL-1β and IL-18 was down-regulated in CO,ACO and KCO groups (P＜0.05).Compared with CO group,neuronal pyroptosis rate and ROS content were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in ACO and KCO groups,and opening of mPTP was significantly inctreased in ACO group (P＜0.05).Conclusion CO postconditioning can inhibit OGD/R-induced pyroptosis in rat hippocampal neurons,and the mechanism is related to inhibiting mPTP/ROS signaling pathway.
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Objective To investigate hyperpolarization-activated cyclic nucleotide-gated (HCN) channels inhibited by dezocine when primary rat cortical astrocytes were exposed to β amyloid peptide.Methods Cultured primary cortical astrocytes from new-born SD rats (within 24 h) were divided into 7 groups (n=5) according to random number table.The astrocytes in groups ZA and CDA were pretreated with 7-CH-cAMP (30 mmol/L),an agonist of HCN channels and ZD7288 (30 mmol/L),an inhibitor of HCN channels for 24 h,respectively.And then the cells in groups DA1,DA2,DA3 and CDA were treated with dezocine 10-7,10-6,10-5and 10-16 mmol/L for 24 h,respectively.Following with the treatments above,the cells in groups A,DA1,DA2,DA3,ZA and CDA were exposed to Aβ1-42 (15μmol/L) for 24 h,but the cells in group C were cultured normally.The effects of cell apoptosis,viability and injury were assessed by Annexin V-FITC/PI assay,MTT assay and lactate de hydrogenase (LDH) release assay.IL-1β was assessed by ELISA assay.The expressions of Cu/ZnSOD and Mn-SOD protein were assessed by Western blot assay.Results Compared with group C,there were significant increases of cell apoptosis,injury and IL-1β level in group A (P＜0.05).Compared with group A,there were significant decreases of cell apoptosis,injury,and IL-1β level but increase of Cu/Zn-SOD and Mn-SOD expressions in groups ZA,DA2 and DA3 (P＜0.05),while the neuroprotection of dezocine was partially restored by 7-CH-cAMP in group CDA (P ＜ 0.05).Conclusion The neuroprotection of dezocine against apoptosis induced by β amyloid peptide could be associated with up-regulation of anti-oxidative stress related Cu/Zn-SOD and Mn-SOD mediated by inhibition of HCN channels.
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Hepatocellular carcinoma (HCC) has high incidence and mortality rates and poor prognosis and greatly threatens human health .Early identification and diagnosis is an important method for reducing the incidence and mortality rates of HCC .The sensitivities and specificities of commonly used serum tumor markers cannot meet the need in clinical practice .Metabolomics focuses on the changes in metabolites produced by organisms, such as lipids, bile acid, and amino acids, which are located at the downstream of systems biology and can directly reflect the biochemical status of tissues .With the help of the techniques including chromatography , mass spectrometry, and magnetic resonance,metabolomics can identify the differences in metabolites in serum , urine, and tissue samples, evaluate their diagnostic values, and help to find potential markers for HCC.In the early screening of tumors, metabolomics has the advantages of easily available specimens ,simple operation, and processing a large number of specimens simultaneously .This article reviews the recent research advances in metabolic markers for HCC with the application of metabolomics.
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Objective To evaluate the role of mitochondrial ATP-seusitive potassium (mito-KATP) channels in sevoflurane postconditioning-induced inhibition of oxygen-glucose dcprivation and restoration (OGD/R)-induced pyroptosis in primary rat cardiomyocytes.Methods Cardiomyocytes of newborn Sprague-Dawley rats (＜48 h after birth) were cultured in vitro and seeded in 6-well dishes (2 cm in diameter)or in 96-well plates.The cells were divided into 6 groups (n =15 each) using a random number table:control group (group C),OGD/R group (group O),sevoflurane postconditioning group (group Sev),sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD) group (group SH),5-HD group (group H) and OGD/R plus 5-HD group (group HO).The cardiomyocytes were subjected to oxygen-glucose deprivation for 4 h followed by restoration of oxygen-glucose supply for 24 h.After oxygen-glucose restoration,the cardiomyocytes in the culture media were exposed to 2％ sevoflurane for 1 h to perform sevoflurane postconditioning.At 1 h before oxygen-glucose deprivation,a specific mito-KATP channel blocker 5-HD 100 μmol/L was added to the culture media.Cardiomyocytes were cultured in normal culture atmosphere in group C.Cardiomyocytes were collected at 24 h of oxygen-glucose restoration.Cell pyroptosis was detected by double flow cytometry AlexaFour488 (caspase-1 FLICA staining) and TMR red (DNA staining) staining.The pyroptosis rate was calculated.The cell survival rate was measured by methyl thiazolyl tetrazolium assay.The content of reactive oxygen species (ROS) in mitochondria was determined by 2',7'-dichlorofluorescin diacetate assay.The mitochondrial membrane potential (MMP) was measured by using JC-I fluorescent probe.The expression of interleukin-1beta (IL-1β) was determined by Western blot.Results Compared with group C,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group O (P＜0.05).Compared with group O,the pyroptosis rate and ROS content were significantly decreased,the cell survival rate and MMP were increased,and the expression of IL-1β was down-regulated in group Sev (P＜0.05).Compared with group Sev,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and M MP were decreased,and the expression of IL-1β was up-regulated in group SH (P＜0.05).Compared with group SH,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group H O (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits OGD/R-induced pyroptosis in primary rat cardiomyocytes is probably associated with increasing mito-KATP channel opening.
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Objective To evaluate the role of extracellular signal-regulated kinase (ERK) signaling pathway in inhibition of oxygen-glucose deprivation and restoration (OGD/R)-induced apoptosis in rat cortical neurons by sevoflurane and the relationship with mitochondrial permeability transition pore (mPTP).Methods The rat cortical neurons were cultured in vitro and seeded in 6-well or 12-well culture plates.The neurons were randomly divided into 4 groups (n =18 each) using a random number table:control group (group C);OGD/R group (group O);sevoflurane group (group OS);sevoflurane + ERK1/2 inhibitor PD98059 group (group OSP).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h in group O.The neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OS.OGD/R was performed at 1 h after ERK1/2 inhibitor PD98059 30 μmol/L was added,and the neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OSP.At 24 h of restoration of O2-glucose supply,the expression of phosphorylated ERK1/2 (p-ERK1/2) in neurons was measured by Western blot,the neuronal apoptosis was detected using Annexin V-FITC/PI double staining combined with flow cytometry,and the opening of mPTP was determined through measuring the optical density at 540 nm.The apoptosis rate was calculated.Results Compared with group C,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly increased in group O (P＜0.05).Compared with group O,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly decreased in group OS (P＜0.05).Compared with group OS,the expression of p-ERK1/2 in neurons was significantly down-regulated,and the apoptosis rate and mPTP opening were significantly increased in group OSP (P＜0.05).Conclusion The mechanism by which sevoflurane inhibits OGD/R-induced apoptosis in rat cortical neurons is related to inhibition of mPTP opening after activation of ERK signaling pathway.
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Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.
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Objective To explore the relationship of sevoflurane neurotoxicity with the expres-sion of Bid,Bim,Puma.Methods The cortical neuron from newborn SD rat (within 24 h)were see-ded in 6 or 12 well plate,and then randomly divided into 4 groups.Rat culture cortical neurons in vitro exposed in 1%,2%,4% and 0% sevoflurane for 6h were divided into A,B,C and D group. The effect of neuron viability,death and apoptosis were assessed using CCK-8,LDH and caspase-3 cleavage 1 7kDa expression assay.The expressions of Bid,Bim and Puma were assessed by western blot.Results Compared with group D, there were significant increases of neuron death and apoptosis,but a decrease of neuron viability,and upregulated expressions of Bid,Bim and Puma in group B (P <0.05);Compared with group B,Group C had increased death and apoptosis and de-creased viability of neurons,as well as upregulated expressions of Bid,Bim and Puma (P <0.05 ). Conclusion Along with the increase of the concentration,sevoflurane neurotoxicity was increased by upregulation of Bid,Bim,Puma expression.
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Objective To explore the mediating role of self-esteem between affective property and metal healthy,and then search effective way to ascent juniors' metal healthy lever.Methods 300 juniors were chosen and received PANAS,MSSMHS and SES assessment.Results ①In sequence,study stress,lability of affect,obsession,anxiety and sensitive of interpersonal relationship produced effect on juniors.②There was a significant negative correlation (r=-0.498～-0.314,P＜0.05)between mental health and self-esteem,positive emotions.There was a significant positive correlation(r=0.291～ 0.520,P＜ 0.05) between mental health and negative emotions.There was a significant positive correlation(r=0.337,P＜0.05)between self-esteem and positive emotions.③The degree of self-esteem and positive emotions had an obvious negatively predictable (R 2=0.223) function to the total average of mental health,and the degree of negative emotions had an obvious positively predictable (R 2=0.350) function to the total average of mental health.④ The mediating effect of subordinates' perceived self-esteem between positive emotions and mental health was also validated(ab/c'=0.118) and the negative emotion had no regulation effect on mental health.Conclusion With the help of significant mediating role of self-esteem,affective property can be seen as a predictors of metal healthy of juniors.
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Objective To evaluate the role of mitochondrial permeability transition pore (mPTP) in sevoflurane-induced inhibition of apoptosis induced by oxygen-glucose deprivation and restoration (OGD/ R) in rat neurons.Methods Cortical neurons isolated from neonatal Sprague-Dawley rats born within 24 h, were cultured primarily and seeded in 6-well plates (2 ml/well) at a density of 1 × 106 cells/ml.The neurons were randomly divided into 5 groups (n =72 each) using a random number table: control group (group C), OGD/R group (group O), mPTP opener group (group A), mPTP opener + sevoflurane group (group AS) , and sevoflurane group (group Sev).The cells were cultured in normal culture medium in group C, and the cells were subjected to OGD for 90 min followed by restoration of O2-glucose supply for 24 h in the other groups.In A and AS groups, mPTP opener atractyloside 20 μmol/L was added immediately after OGD.In Sev and AS groups, the cells were post-conditioned with 2％ sevoflurane for 1 h immediately after OGD.The neurons were collected at 24 h of OGD in O and A groups, or at 23 h of OGD in Sev and AS groups.Annexin V-FITC/PI double staining was performed to count apoptotic neurons, apoptotic rate was calculated, and cell survival rate was measured using MTT assay.The mitochondrial membrane potential (MMP) was measured by using JC-1 fluorescence probe as indicator.The opening of mPTP was determined through assessing the changes of mitochondrial optical density (△OD540 of mPTP).The expression of pro-apoptotic factors Bid, Bim, Puma protein and mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction and Western blot.Results Compared with group C, apoptotic rate was significantly increased, cell survival rate and MMP were decreased, the opening of mPTP was increased, and the expression of Bid, Bim, Puma protein and mRNA was up-regulated in the other groups.Compared with group O, apoptotic rate was significantly decreased, cell survival rate and MMP were increased, the opening of mPTP was decreased, and the expression of Bid, Bim, Puma protein and mRNA was down-regulated in Sev group.Compared with group Sev, apoptotic rate was significantly increased,cell survival rate and MMP were decreased, the opening of mPTP was increased, and the expression of Bid, Bim, Puma protein and mRNA was up-regulated in A and As groups.Compared with group A, apoptotic rate was significantly decreased, cell survival rate and MMP were increased, the opening of mPTP was decreased, and the expression of Bid, Bim, Puma protein and mRNA was down-regulated in group AS.Conclusion Sevoflurane mitigates OGD/R-induced apoptosis in rat neurons through inhibiting mPTP opening.