OBJECTIVE To explore the clinical efficacy of Fuxin Mixture in the treatment of patients with heart failure of yang deficiency and water inundation type after myocardial infarction and its effect on serum myocardial injury and ventricular remodeling re-lated indicators.METHODS A total of 92 patients with heart failure of yang deficiency and water inundation type after myocardial infarction were randomly divided into an observation group and a control group(n=46).The control group was given basic western medicine treatment,and the observation group was given Fuxin Mixture on the basis of the control group's treatment.The TCM syn-drome scores,ultrasound cardiogram indices[left ventricular ejection fraction(LVEF),left ventricular Tei index,and stroke volume(SV)],serum myocardial injury indices[serum N-terminal pro-B-type natriuretic peptide(NT-ProBNP)level,serum high-sensitiv-ity cardiac troponin I(hs-cTnI)level,and creatine kinase isoenzyme(CK-MB)level],serum ventricular remodeling indices[hypoxi-a-inducible factor 1α(HIF-1α),soluble growth-stimulated gene expressed 2 protein(sST2),and angiotensin Ⅱ(AngⅡ)],and changes in the Minnesota Living with Heart Failure Questionnaire(MLHFQ)were evaluated before and after treatment.The two groups were followed up for 1 year after treatment to observe the occurrence of major adverse cardiovascular events.RESULTS After treat-ment,the TCM syndrome scores of both groups were significantly reduced(P＜0.01),the observation group was better than the control group(P＜0.05,P＜0.01),and the total effective rate of TCM in the observation group was better than that of the control group(P＜0.05);the serum ultrasound cardiogram indexes,myocardial injury,ventricular remodeling indexes and MLHFQ scores of the two groups were significantly reduced(P＜0.01),and the observation group was better than the control group(P＜0.01).During the fol-low-up period,the all-cause mortality,malignant arrhythmia rate and acute heart failure rate of the observation group after treatment were significantly lower than those of the control group(P＜0.05,P＜0.01).CONCLUSION Fuxin Mixture has significant clinical efficacy in the treatment of patients with heart failure of yang deficiency and water inundation type after myocardial infarction,and the efficacy mechanism may be related to its reduction of myocardial injury and inhibition of ventricular remodeling.

Objective:To evaluate the effects of endothelial progenitor cell (EPC)-derived exosomes on neuronal injury induced by oxygen-glucose deprivation and restoration (OGD/R).Methods:HT22 neurons of mice were cultured and divided into 3 groups ( n=30 each) using a random number table method: control group (C group), OGD/R group and OGD/R plus EPC-derived exosome group (OGD/R+ EXO group). Cells in group C were cultured in normal atmosphere.In group OGD/R, the cells were exposed to 94%N 2-1%O 2-5%CO 2 for 6 h in glucose- and serum-free DMEM medium, followed by 24 h restoration of O 2 and glucose in the normal medium.In group OGD/R+ EXO, 20 μg/ml EPC-derived exosomes were added to the culture medium at 24 h before developing the model.EPCs were identified by immunofluorescence staining.Exosomes were identified by Western blot, transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Cell viability was measured by CCK-8 assay, the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by enzyme-linked immunosorbent assay.The neuronal apoptosis was detected by TUNEL staining, and the apoptosis rate was calculated.The expression of Bax, Bcl-2, caspase-3, and cleaved caspase-3 was determined by Western blot, and cleaved caspase-3/caspase-3 ratio was calculated. Results:The cultured cells were EPCs, and EPC-derived exosomes were successfully extracted.Compared with group C, the cell viability was significantly decreased, the content of MDA was increased, the activity of SOD was decreased, the apoptosis rate was increased, the expression of Bax was up-regulated, the expression of Bcl-2 was down-regulated, and the ratio of cleaved-caspase-3/caspase-3 was increased in group OGD/R and group OGD/R+ EXO ( P<0.05). Compared with group OGD/R, the cell viability was significantly increased, the content of MDA was decreased, the activity of SOD was increased, the apoptosis rate was decreased, the expression of Bax was down-regulated, the expression of Bcl-2 was up-regulated, and the ratio of cleaved-caspase-3/caspase-3 was decreased in group OGD/R+ EXO ( P<0.05). Conclusions:EPC-derived exosomes can reduce OGD/R-induced neuronal injury, which is related to inhibition of oxidative stress and neuronal apoptosis.

Objective:To evaluate the role of hemopexin (HPX) in cerebral ischemia-reperfusion (I/R) injury in rats.Methods:One hundred and twenty healthy male Sprague-Dawley rats, aged 7-8 weeks, weighing 250-280 g, were divided into sham operation group (S group, n=36), cerebral I/R group (I/R group, n=36), vehicle group (V group, n=24), and HPX group ( n=24). The model of cerebral I/R injury was established by 120 min middle cerebral artery occlusion followed by reperfusion in anesthetized rats.At 6, 12 and 24 h of reperfusion, 4 rats in S group and I/R group were sacrificed, and the ischemic penumbra of the ipsilateral cerebral cortex was obtained to detect the expression of HPX by Western blot.In I/R, V and HPX groups, 0.9% normal saline 10 μl, 0.1% NaN 3 10 μl, and 1.86 mg/ml HPX 10 μl were injected into the lateral ventricle, respectively, immediately after reperfusion.Eight rats in each group were selected, and neurological deficit was scored at 1-7 days of reperfusion.Eight rats in each group were sacrificed at 1 and 7 days of reperfusion, brains were removed, and brain tissues were obtained for measurement of infarct size, and the percentage of infarct size was calculated. Results:Compared with S group, the expression of HPX in cerebral ischemic penumbra was significantly up-regulated at 24 h of reperfusion in I/R group, and the neurological deficit scores were significantly decreased at 1-7 days of reperfusion, and the percentage of cerebral infarct size was increased at 1 and 7 days of reperfusion in I/R, V and HPX groups ( P<0.05). Compared with I/R group, the neurological deficit scores were significantly increased at 1-7 days of reperfusion, and the percentage of cerebral infarct size was decreased at 1 and 7 days of reperfusion in HPX group ( P<0.05), and no significant change was found in the above indicators in V and I/R groups ( P>0.05). Conclusion:Up-regulation of HPX expression is the endogenous protective mechanism of cerebral I/R injury in rats.

Background: Diabetes-related neuropathic pain frequently occurs, and the underpinning mechanism remains elusive. The periaqueductal gray (PAG) exhibits descending inhibitory effects on central pain transmission. The current work aimed to examine whether inflammatory cytokines regulate mechanical allodynia and thermal hyperalgesia induced by diabetes through the phosphoinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathway in the PAG. Methods: Streptozotocin (STZ) was administered intraperitoneally to mimic allodynia and hyperalgesia evoked by diabetes in rats. Behavioral assays were carried out for determining mechanical pain and thermal hypersensitivity. Immunoblot and ELISA were performed to examine PAG protein amounts of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), as well as their corresponding receptors in STZ rats, and the expression of PI3K/protein kinase B (Akt)/mTOR signaling effectors. Results: Increased PAG p-PI3K/p-Akt/p-mTOR protein amounts were observed in STZ-induced animals, a PI3K-mTOR pathway inhibition in the PAG attenuated neuropathic pain responses. Moreover, the PAG concentrations of IL-1β, IL-6, and TNF-α and their receptors (namely, IL-1R, IL-6R, and tumor necrosis factor receptor [TNFR] subtype TNFR1, respectively) were increased in the STZ rats. Additionally, inhibiting IL-1R, IL-6R, and TNFR1 ameliorated mechanical allodynia and thermal hyperalgesia in STZ rats, alongside the downregulation of PI3K-mTOR signaling. Conclusions Overall, the current study suggests that upregulated proinflammatory cytokines and their receptors in the PAG activate PI3K-mTOR signaling, thereby producing a de-inhibition effect on descending pathways in modulating pain transmission, and eventually contributing to neuropathic pain.

Objective:To evaluate the effect of acetaminophen on sepsis-associated encephalopathy (SAE) in mice and the relationship with ferroptosis.Methods:A total of 160 clean-grade healthy adult male C57BL/6J mice, aged 6 weeks, weighing 22-24 g, were divided into 4 groups ( n=40 each) using a random number table method: sham operation group (Sham group), sham operation+ acetaminophen group (Sham+ APAP group), SAE group and SAE+ acetaminophen group (SAE+ APAP group). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Acetaminophen 100 mg/kg was intraperitoneally injected at 1 h before the model was established in group Sham+ APAP and group SAE+ APAP.The postoperative 7-day survival rate was recorded.At 24 h after operation, brain tissues were taken for examination of the pathological changes of neurons in hippocampal CA1 region.At 24 h after establishment of the model, ultrastructure was observed with a transmission electron microscope, the contents of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) in hippocampus were determined by colorimetry, and the expression of glutathione peroxidase (GPX4), cystine/glutamate antiporter (xCT) and 4-hydroxy-2-nonenal (4-HNE) was determined by Western blot. Results:Compared with group Sham, the postoperative 7-day survival rate of mice was significantly decreased, contents of ROS and MDA in hippocampus were increased, GSH content in hippocampus was decreased and expression of GPX4 and xCT was down-regulated in SAE and SAE+ APAP groups, 4-HNE expression was up-regulated in group SAE ( P<0.05), and no significant change was found in the parameters mentioned above in group Sham+ APAP ( P>0.05). Compared with group SAE, the postoperative 7-day survival rate of mice was significantly increased, contents of ROS and MDA in hippocampus were decreased, GSH content in hippocampus was increased, expression of GPX4 and xCT was up-regulated and expression of 4-HNE was down-regulated in group SAE+ APAP ( P<0.05). The pathological changes in hippocampal CA1 region and damage to ultrastructure of hippocampal tissue were significantly attenuated in group SAE+ APAP as compared with group SAE. Conclusion:Acetaminophen can effectively alleviate SAE, and the mechanism is related to inhibiting ferroptosis in mice.

Objective:To evaluate the relationship between phosphorylation of Tau protein and apolipoprotein E (ApoE) containing 18 kDa fragments and investigate the mechanism of neuronal damage induced by sevoflurane.Methods:Primary neurons (ApoE3 and ApoE2 genotypes, 24 dishes for each genotype) of fetal mice cultured until the 5th day were divided into 4 groups ( n=12 each) using a random number table method: ApoE3 control group (A3C group), ApoE3 sevoflurane group (A3S group), ApoE2 control group (A2C group) and ApoE2 sevoflurane group (A2S group). Neurons were treated with 21% oxygen + 5% carbon dioxide + 4.1% sevoflurane for 4 h in A3S and A2S groups, while the neurons were only treated with 21% oxygen + 5% carbon dioxide in A3C and A2C groups.The cell proteins were then extracted to detect the expression of full-length ApoE and ApoE, AT8 and PHF1 containing 18 kDa fragments (by Western blot), expression of ApoE mRNA (by real-time polymerase chain reaction), and concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant (by enzyme-linked immunosorbent assay). Results:Compared with A2C group, the expression of ApoE mRNA and full-length ApoE in neurons was up-regulated ( P<0.05), and no significant change was found in the expression of AT8 and PHF1 and concentrations of TNF-α and IL-6 in the supernatant in A2S group ( P>0.05). Compared with A3C group, the expression of ApoE mRNA, full-length ApoE, and ApoE, AT8 and PHF1 containing 18 kDa fragments was up-regulated, and the concentrations of TNF-α and IL-6 in the supernatant were increased in A3S group ( P<0.05). Conclusion:Sevoflurane may promote phosphorylation of Tau proteins and increase inflammatory responses through up-regulating the expression of ApoE containing 18 kDa fragments, thus leading to neuronal damage.

Background: Diabetes-related neuropathic pain frequently occurs, and the underpinning mechanism remains elusive. The periaqueductal gray (PAG) exhibits descending inhibitory effects on central pain transmission. The current work aimed to examine whether inflammatory cytokines regulate mechanical allodynia and thermal hyperalgesia induced by diabetes through the phosphoinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathway in the PAG. Methods: Streptozotocin (STZ) was administered intraperitoneally to mimic allodynia and hyperalgesia evoked by diabetes in rats. Behavioral assays were carried out for determining mechanical pain and thermal hypersensitivity. Immunoblot and ELISA were performed to examine PAG protein amounts of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), as well as their corresponding receptors in STZ rats, and the expression of PI3K/protein kinase B (Akt)/mTOR signaling effectors. Results: Increased PAG p-PI3K/p-Akt/p-mTOR protein amounts were observed in STZ-induced animals, a PI3K-mTOR pathway inhibition in the PAG attenuated neuropathic pain responses. Moreover, the PAG concentrations of IL-1β, IL-6, and TNF-α and their receptors (namely, IL-1R, IL-6R, and tumor necrosis factor receptor [TNFR] subtype TNFR1, respectively) were increased in the STZ rats. Additionally, inhibiting IL-1R, IL-6R, and TNFR1 ameliorated mechanical allodynia and thermal hyperalgesia in STZ rats, alongside the downregulation of PI3K-mTOR signaling. Conclusions Overall, the current study suggests that upregulated proinflammatory cytokines and their receptors in the PAG activate PI3K-mTOR signaling, thereby producing a de-inhibition effect on descending pathways in modulating pain transmission, and eventually contributing to neuropathic pain.

Objective:To evaluate the effectiveness and safety of early exercise safety management in patients undergoing mechanical ventilation in intensive care unit (ICU).Methods:A prospective historical control observation was conducted. Forty-five patients with severe respiratory failure undergoing mechanical ventilation admitted to the ICU of Affiliated Hospital of Jining Medical University from April to June in 2019 were enrolled in the observation group and implemented early exercise safety management, including establishing multidisciplinary safety management team with ICU doctors, ICU nurses, respiratory therapists, rehabilitation therapists, dietitians and psychological consultants to jointly develop early exercise plan; equipping with sports and safety protection equipment; assessing the early exercise risks, formulating early exercise prescriptions; formulating the nutritional and psychological prescriptions; carrying out the propaganda and education in the early exercise process of patients and communicating with patients timely; strengthening team training. Other 45 patients with severe respiratory failure admitted for mechanical ventilation from January to March in 2019 were enrolled in the control group, whose gender, age and artificial airway conditions were matched with those in the observation group. Routine mechanical ventilation nursing was performed, including condition monitoring, medication nursing, environmental management and routine examination assistance. The improvement of depression, anxiety, comfort, lung function, and quality of life before and after nursing intervention were observed, and the occurrence of complications was recorded.Results:There was no significant difference in gender, age or artificial airway conditions between the two groups. Before nursing intervention, there was no significant difference in depression, anxiety, comfort, lung function and quality of life between the two groups. After the intervention of different nursing programs, the above conditions of the two groups were significantly improved, and the improvement was more significant in the observation group, which showed that the hospital anxiety and depression scale (HAD) score and forced vital capacity (FVC) of the observation group were significantly lower than those of the control group [HAD score: 10.80±2.54 vs. 17.51±3.66, FVC (L): 1.81±0.42 vs. 2.23±0.39, both P < 0.01], while the general comfort questionnaire (GCQ) score, forced expiratory volume in 1 second (FEV1), FEV1/FVC ratio, FEV1 percentage of predicted (FEV1%) and each dimension score of 36-item short form health survey (SF-36) scale were significantly higher than those of the control group [GCQ score: 110.87±5.33 vs. 96.93±3.02, FEV1 (L): 1.99±0.37 vs. 1.71±0.15, FEV1/FVC ratio: 0.88±0.04 vs. 0.84±0.03, FEV1%: (88.98±8.57)% vs. (80.41±4.45)%, mental function score: 49.74±9.88 vs. 40.17±8.95, physical function score: 27.65±9.46 vs. 20.32±9.53, social relationship score: 62.14±6.33 vs. 50.28±8.76, general health score: 38.61±8.48 vs. 30.63±8.93, all P < 0.01]. The overall incidence of complications in the observation group was significantly lower than that in the control group (24.44% vs. 57.78%, P < 0.01). Conclusions:Early exercise safety management scheme for patients undergoing mechanical ventilation in ICU can improve clinical efficacy and reduce complications. At the same time, it can further standardize the behaviors of medical staff and ensure the early exercise.

Objective:To evaluate the effect of hydrogen on lipopolysaccharide (LPS)-caused inflammatory responses in BV-2 microglia and the role of autophagy.Methods:The BV-2 microglial cells cultured in vitro were seeded in 6- or 96-well plates and were divided into 4 groups ( n=24 each) using a random number table method: control group (group C), group LPS, hydrogen-rich medium group (group H) and autophagy inhibitor 3-methylpurine group (group 3-MA). In group C, cells were cultured in MEM culture medium supplemented with 15% fetal bovine serum for 24 h. In group LPS, LPS was added at a final concentration of 1 μg/ml, and cells were incubated for 24 h. In group H, LPS was added at a final concentration of 1 μg/ml, the culture medium was replaced with a hydrogen-rich medium at a final concentration of 0.6 mmol/L, and cells were incubated for 24 h. In group 3-MA, 3-methylpurine was added at a final concentration of 2 mmol/L, and the subsequent treatment was similar to those previously described in group H. The cell survival rate was detected by CCK-8 assay.The concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and transforming growth factor-β (TGF-β) in supernatant were detected by enzyme-linked immunosorbent assay.The percentage of ionized calcium binding adaptor molecule-1 (Iba-1) +, Iba-1 + CD86 + and Iba-1 + CD206 + cells was detected by flow cytometry.The expression of microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ), LC3Ⅱ, Beclin-1 and p62 was detected by Western blot, and the ratio of LC3Ⅱ/LC3Ⅰ was calculated. Results:There was no significant difference in the cell survival rate among the four groups ( P>0.05). Compared with group C, the concentrations of TNF-α, IL-6, IL-10 and TGF-β and percentage of Iba-1 +, Iba-1 + CD86 + and Iba-1 + CD206 + cells were significantly increased in LPS, H and 3-MA groups, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was significantly down-regulated, and p62 expression was up-regulated in LPS and 3-MA groups, and the ratio of LC3LC3Ⅱ/LC3Ⅰ and Beclin-1 expression was significantly up-regulated, and p62 expression was down-regulated in group H ( P<0.05). Compared with group LPS, the concentrations of TNF-α and IL-6 were significantly decreased, the concentrations of IL-10 and TGF-β were increased, the percentage of Iba-1 + and Iba-1 + CD86 + cells were decreased, the percentage of Iba-1 + CD206 + cells was increased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was up-regulated, and p62 expression was down-regulated in group H ( P<0.05), and no significant change was found in the above indexes in group 3-MA ( P>0.05). Compared with group H, the concentrations of TNF-α and IL-6 were significantly increased, the concentrations of IL-10 and TGF-β were decreased, the percentage of Iba-1 + and Iba-1 + CD86 + cells was increased, the percentage of Iba-1 + CD206 + cells was decreased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was down-regulated, and p62 expression was up-regulated in group 3-MA ( P<0.05). Conclusion:The mechanism by which hydrogen reduces LPS-caused inflammatory responses in BV-2 microglia is related to enhancing autophagy and inhibiting microglial activation.

Objective:To evaluate the effect of dexmedetomidine on intestinal injury in severely burned rats.Methods:One hundred and twenty healthy clean-grade male Sprague-Dawley rats, weighing 240-260 g, were divided into 4 groups ( n=30 each) using a random number table method: sham operation group (group Sham), sham plus dexmedetomidine group (group Sham+ Dex), severe burn group (group Burn), and severe burn plus dexmedetomidine group (group Burn+ Dex). Forty percent total body surface area of III degree burn model was developed in chloral hydrate-anesthetized rats.Dexmedetomidine was intravenously infused for 4 h at a rate of 5 μg·kg -1·h -1 starting from 3 h after establishing the model in Sham+ Dex group and Burn+ Dex group.The small intestinal tissues were removed for examination of the pathological changes which were scored and for determination of tumor necrosis factor-alpha (TNF-α) and high-mobility group box 1 protein (HMGB1) contents (by enzyme-linked immunosorbent assay), and expression of occludin and ZO-1 protein (by Western blot). The serum concentrations of 4-kD-FITC were measured at 90, 180, 360 and 720 min after establishing the model. Results:Compared with Sham group, the pathological scores of intestinal tissues, contents of TNF-α and HMGB1, serum concentrations of 4-kD-FITC at each time point were significantly increased, and the expression of occludin and ZO-1 was down-regulated in Burn group and Burn+ Dex group ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ Dex group ( P>0.05). Compared with Burn group, the pathological scores of intestinal tissues, contents of TNF-α and HMGB1, serum concentrations of 4-kD-FITC at each time point were significantly decreased, and the expression of occludin and ZO-1 was up-regulated in Burn+ Dex group ( P<0.05). Conclusion:Dexmedetomidine can reduce intestinal injury in severely burned rats, and the mechanism may be related to inhibiting inflammatory responses in the intestine.