Role of IL-6 trans-signaling pathway in perioperative neurocognitive disorder in mice / 中华麻醉学杂志

Objective:To investigate the role of IL-6 trans-signaling pathway in perioperative neurocognitive disorder in mice.Methods:Eighty-four SPF healthy male C57BL/6 wild-type mice and 84 SPF healthy male IL-6R -/- mice, aged 12-14 weeks, weighing 25-35 g, were used.The 84 wild-type mice were divided into 4 groups ( n=21 each) using a random number table method: sham group (SH group), surgery group (S group), sgp130Fc (specific IL-6 trans-signaling pathway blocker) group (F group), and sgp130Fc+ surgery group (FS group). In S group and FS group, internal fixation was performed under general anesthesia with sevoflurane after tibial fracture.Mice only received anaesthesia with sevoflurane in SH group and F group.In FS group and F group, sgp130Fc 10 mg/kg was intraperitoneally injected before anesthesia.Blood samples were collected from the celiac vein at 24 h after surgery for determination of the concentrations of interleukin 6 (IL-6), IL-1β and tumor necrosis factor (TNF)-α in plasma by enzyme-linked immunosorbent assay (ELISA). Then the mice were sacrificed, brains were removed, and hippocampal tissues were obtained for measurement of the contents of IL-6, IL-1β and TNF-α (by ELISA) and for observation of activation of microglias in the hippocampal DG region (by immunofluorescence staining, n=6). Cognitive function was evaluated by contextual fear conditioning test ( n=15) on 3 days after surgery.Eighty-four IL-6R -/- mice were randomly divided into 4 groups ( n=21 each): sham group (KO-SH group), surgery group (KO-S group), saline group (KO-C group), and hyper IL-6 (specific IL-6 trans-signaling pathway activator) group (KO-H group). The treatment in KO-SH group and KO-S group was the same as those previously described in SH group and S group, respectively.0.9% NaCl solution 100 μl was intraperitoneally injected in KO-C group, 100 μl hyper IL-6 40 μg/kg was intraperitoneally injected in KO-H group, and 24 h later blood was collected from the celiac vein for measurement of the concentrations of IL-6, IL-1β and TNF-α in plasma by ELISA.Then the mice were sacrificed, brains were removed, and hippocampal tissues were obtained for determination of the contents of IL-6, IL-1β and TNF-α (by ELISA) and for observation of activation of microglias in the hippocampal DG region (by immunofluorescence staining, n=6). Cognitive function was evaluated by contextual fear conditioning test ( n=15) on 3 days after surgery. Results:Compared with SH group, the percentage of freezing time in the contextual fear conditioning test was significantly decreased, and the activation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampi were increased in S group ( P<0.05). Compared with S group, the percentage of freezing time in the contextual fear conditioning test was significantly increased, and the activation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampus were decreased in FS group ( P<0.05). There were no significant differences in the percentage of freezing time, activation of microglias in the hippocampal DG region, and levels of IL-6, IL-1β and TNF-α in plasma and hippocampi between KO-S group and KO-SH group ( P>0.05). Compared with KO-C group, the percentage of freezing time in the contextual fear conditioning test was significantly decreased, and the activiation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampus were increased in KO-H group ( P<0.05). Conclusions:IL-6 trans-signaling pathway is involved in the process of perioperative neurocognitive disorder in mice.

Effect of clemastine fumarate on TLR4/PI3K/Akt signaling pathway during hypoxia-reoxygenation in rat cardiomyocytes / 中华麻醉学杂志

Objective To evaluate the effect of clemastine fumarate on Toll-like receptor 4/phosphatidylinositol-3-kinase/serine-threonine kinase (TLR4/PI3K/Akt) signaling pathway during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9C2 cells of rats cultured in vitro were seeded in culture wells or dishes at a density of 1×105 cells/ml and divided into 3 groups (n=11 each) by using a random number table method:control group (group C),H/R group and clemastine fumarate group (CF group).Cardiomyocytes were exposed to 5％ CO2-95％ N2in a low-glucose DMEM medium at 37℃ for 4 h followed by 4 h reoxygenation.At 4 h of reoxygenation,the cell viability was detected by CCK-8 assay,the ultrastructure was observed with a transmission electron microscope,the expression of TLR4,PI3K,phosphorylated Akt (p-Akt) and caspase-3 was detected by Western blot,and the expression of TLR4,PI3K and caspase-3 was detected by immunofluorescence.Results Compared with group C,the cell viability was significantly decreased,the expression of TLR4 and caspase-3 was up-regulated,and the expression of PI3K and p-Akt was down-regulated in group H/R (P＜0.05).Compared with group H/R,the cell viability was significantly increased,the expression of TLR4 and caspase-3 was down-regulated,the expression of PI3K and p-Akt was up-regulated (P＜0.05),and the mitochondrial damage was significantly attenuated in group CF.Conclusion The mechanism by which clemastine fumarate alleviates H/R injury to rat cardiomyocytes may be related to inhibiting TLR4 expression and activating PI3K/Akt signaling pathway.

Stem Cell-Based Therapies for Liver Diseases: An Overview and Update

BACKGROUND: Liver disease is one of the top causes of death globally. Although liver transplantation is a very effective treatment strategy, the shortage of available donor organs, waiting list mortality, and high costs of surgery remain huge problems. Stem cells are undifferentiated cells that can differentiate into a variety of cell types. Scientists are exploring the possibilities of generating hepatocytes from stem cells as an alternative for the treatment of liver diseases. METHODS: In this review, we summarized the updated researches in the field of stem cell-based therapies for liver diseases as well as the current challenges and future expectations for a successful cell-based liver therapy. RESULTS: Several cell types have been investigated for liver regeneration, such as embryonic stem cells, induced pluripotent stem cells, liver stem cells, mesenchymal stem cells, and hematopoietic stem cells. In vitro and in vivo studies have demonstrated that stem cells are promising cell sources for the liver regeneration. CONCLUSION: Stem cell-based therapy could be a promising therapeutic method for patients with end-stage liver disease, which may alleviate the need for liver transplantation in the future.

A path analysis of impacts of social support and rumination on posttraumatic growth of patients with human immunodeficiency virus / 中国实用护理杂志

Objective To explore the effect of social support and rumination on posttraumatic growth of patients with human immunodeficiency virus (HIV). Methods A total of 1 152 patients with HIV from Shanghai Public Clinical Center were investigated using General questionnaire, Perceived Social Support Scale, Event Related Rumination Inventory and Posttraumatic Growth Inventory by cross-sectional survey method from January 2018 to October 2018. The path of social support and rumination on post-traumatic growth was established by correlation analysis and structural equation model. Results The total score of posttraumatic growth in patients with HIV was (47.93±23.55) points, which was at the low-middle level. Correlation analysis showed that posttraumatic growth was positively correlated with comprehension of social support (r=0.234, P＜0.01), positively correlated with rumination (r=0.352, P＜0.01). Structural equation model showed social support had directly positive effect on posttraumatic growth, path coefficient were 0.55. Rumination had a partial mediating effect between social support and posttraumatic growth, and mediation effects account for 11.65% of the total effect. Conclusions The posttraumatic growth level of patients with HIV needs to be improved. Health care providers should help patients get as high a level of social support as possible, as well as focus on guiding patients to think positive about the disease.

A path analysis of impacts of social support and rumination on posttraumatic growth of patients with human immunodeficiency virus / 中国实用护理杂志

Objective@#To explore the effect of social support and rumination on posttraumatic growth of patients with human immunodeficiency virus (HIV).@*Methods@#A total of 1 152 patients with HIV from Shanghai Public Clinical Center were investigated using General questionnaire, Perceived Social Support Scale, Event Related Rumination Inventory and Posttraumatic Growth Inventory by cross-sectional survey method from January 2018 to October 2018. The path of social support and rumination on post-traumatic growth was established by correlation analysis and structural equation model.@*Results@#The total score of posttraumatic growth in patients with HIV was (47.93±23.55) points, which was at the low-middle level. Correlation analysis showed that posttraumatic growth was positively correlated with comprehension of social support (r=0.234, P<0.01), positively correlated with rumination (r=0.352, P<0.01). Structural equation model showed social support had directly positive effect on posttraumatic growth, path coefficient were 0.55. Rumination had a partial mediating effect between social support and posttraumatic growth, and mediation effects account for 11.65% of the total effect.@*Conclusions@#The posttraumatic growth level of patients with HIV needs to be improved. Health care providers should help patients get as high a level of social support as possible, as well as focus on guiding patients to think positive about the disease.

Effect of sevoflurane preconditioning on expression of metabotropic glutamate receptor typeⅡ dur-ing focal cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of sevoflurane preconditioning on the expression of metabotropic glutamate receptor type Ⅱ( mGluRⅡ) during focal cerebral ischemia-reperfusion ( I∕R) in rats. Methods Forty-eight clean-grade healthy male Sprague-Dawley rats were divided into 3 groups (n＝16 each) using a random number table method: sham operation group ( group S), cerebral I∕R group (group I∕R) and sevoflurane preconditioning group (group Sev). Rats were anesthetized with 10% chloral hydrate 3 ml∕kg. Focal cerebral I∕R was produced by occlusion of the right middle cerebral artery for 2 h fol-lowed by 24 h reperfusion. In group Sev, 2. 7% sevoflurane was inhaled for 1 h and 24 h later focal cerebral I∕R was produced. At 24 h after reperfusion, neurological deficit was scored, the cerebral infarct size was determined by TTC staining, the cell apoptosis in ischemic penumbra was observed by TUNEL, IκB-α ex-pression was detected by Western blot, and mGluRⅡexpression was determined by immunofluorescent stai-ning. The apoptosis rate was calculated. Results Compared with group S, the neurological deficit score, cerebral infarct size and apoptosis rate were significantly increased, the expression of mGluRⅡwas up-regu-lated, and the expression of IκB-α was down-regulated in I∕R and Sev groups ( P＜0. 05). Compared with group I∕R, the neurological deficit score, cerebral infarct size and apoptosis rate were significantly de-creased, the expression of mGluRⅡwas down-regulated, and the expression of IκB-α was up-regulated in group Sev (P＜0. 05). Conclusion Sevoflurane preconditioning reduces focal cerebral I∕R injury through inhibiting the expression of mGluRⅡ in rats.

Effect of isoflurane preconditioning on inflammatory responses during spinal cord injury in rats / 中华麻醉学杂志

Objective To evaluate the effect of isoflurane preconditioning on inflammatory responses during spinal cord injury (SCI ) in rats .Methods Sixty adult male Sprague-Dawley rats ,weighing 250-300 g , were randomly divided into 3 groups ( n= 20 each ) using a random number table :sham operation group (S group) , SCI group , and isoflurane preconditioning group (I group ) . The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg .SCI was produced by a weight-drop contusion at the T10 level .The rats inhaled 2% isoflurane for 2 h ,and the model was established at 24 h after the end of isoflurane inhalation in I group . Neurological function was assessed and scored by using the the Basso , Beattie , Bresnahan (BBB ) Locomotor Rating Scale on 7 days after SCI .Five rats in each group were then chosen and spinal cord specimens were obtained and cut into sections which were stained with haematoxylin and eosin for determination of the viable neuron count .Fifteen rats in each group were sacrificed and the spinal cord was removed for detection of nuclear factor kappaB (NF-κB ) and interleukin-1β (IL-1β) expression (by Western blot ) .Results Compared with S group ,BBB score and the number of viable neurons were significantly decreased ,and the expression of NF-κB and IL-1βprotein was up-regulated in SCI group ( P<0.05) .Compared with SCI group ,BBB score and the number of viable neurons were significantly increased ,and the expression of NF-κB and IL-1βprotein was down-regulated in group I ( P<0.05 ) .Conclusion The mechanism by which isoflurane preconditioning protects the spinal cord is related to inhibition of inflammatory responses in rats .

Inhibitory effect of alantolactone on the proliferation of K562/ADR cells and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the inhibitory effect of alantolactone on the proliferation of adriamycin-resistant human chronic myelogenous leukemia cell line K562/ADR cells and its mechanism.</p><p><b>METHODS</b>K562/ADR cells were treated with various concentrations of alantolactone (0, 1, 2, 4, 6, 8, and 10 μmol/L) for different time points. Cell viability was analyzed with MTT assay. The effect of alantolactone on the apoptosis of K562/ADR cells was measured by flow cytometry. The expression of apoptosis-related proteins after treatment with alantolactone was analyzed using Western blot.</p><p><b>RESULTS</b>Alantolactone could effectively inhibit the proliferation of K562/ADR cells in dose- and time- dependent manner, the IC50 value of alantolactone treatment of K562/ADR cells for 24 h was 4.7 μmol/L (P<0.05). Flow cytometric analysis displayed that the apoptotic rates were 1.35%, 16.91%, 29.61% and 46.26%, respectively, after treatment with alantolactone at 0, 2.5, 5 and 7.5 μmol/L. Meanwhile, the expression of Bcl-2 and BCR-ABL proteins were significantly decreased and that of Bax, cytochrome C, cleaved-caspase-9, cleaved-caspase-3 and cleaved-PARP increased by alantolactone treatment.</p><p><b>CONCLUSION</b>Alantolactone had obvious inhibitory effect on the proliferation of K562/ADR cells through the caspase dependent mitochondrial(or intrinsic)apoptotic pathway.</p>

Role of δ opioid receptor in brain injury following asphyxial cardiac arrest-resuscitation in rats / 中华麻醉学杂志

Objective To evaluate the role of δ opioid receptor in the brain injury following asphyxial cardiac arrest-resuscitation in rats.Methods Ninety-six pathogen-free male Sprague-Dawley rats,weighing 300-350 g,were randomly divided into 4 groups (n =24 each):sham operation group (group S),asphyxial cardiac arrest-resuscitation group (group M),δ opioid receptor agonist BW373U86 group (group B) and δ opioid receptor antagonist naltrindole group (group N).Cardiac arrest was induced by clamping the tracheal tube for 8 min.Mechanical ventilation with pure oxygen was performed.Epinephrine 0.02 mg/kg and 5％ NaHCO3 1 mg/kg were injected intravenously as soon as chest compression was started.Appearance of spontaneous breathing and MAP ＞ 50 mm Hg (lasting for more than 10 min) were considered to be signs of successful recovery of spontaneous circulation (ROSC).BW373U86 and naltrindole 1 mg/kg were injected via the femoral vein immediately after ROSC in groups B and N,respectively,while the equal volume of normal saline was given instead in groups S and M.Neurological deficit score (NDS) was evaluated at 3,24 and 72 h after ROSC.The rats were then sacrificed,brains were isolated and the hippocampus was obtained for detection of the expression of brain-derived neurotrophic factor (BDNF)and tyrosine receptor kinase B (TrkB)mRNA by RT-PCR.The histological grading (HG) of neurons and number of survival neurons in hippocampal CA1 region were determined at 72 h after ROSC.Results Compared with group S,the expression of BDNF and TrkB mRNA was significantly up-regulated,HG was increased,and NDS and the number of survival neurons were decreased in groups M,B and N (P ＜ 0.05).Compared with group M,the expression of BDNF and TrkB mRNA was significantly up-regulated in group B,the expression of BDNF and TrkB mRNA was down-regulated in group N,and HG was significantly decreased,and NDS and the number of survival neurons were increased in groups B and N (P ＜ 0.05).NDS was significantly lower,the number of survival neurons was smaller,the expression of BDNF and TrkB mRNA was lower,and HG was higher in group N than in group B (P ＜ 0.05).Conclusion Activation of δ opioid receptor can reduce the brain injury following asphyxial cardiac arrest-resuscitation in rats and the mechanism may be related to up-regulation of BDNF and TrkB after activation of δ opioid receptor.

Effect of isoflurane preconditioning on expression of 5-1ipoxygenase during focal cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To investigate the effect of isoflurane preconditioning on the expression of 5-lipoxy-genase (5-LOX) during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-nine male adult Sprague-Dawley rats weighing 250-300 g were randomly divided into 3 groups (n =13 each):sham operation group (group S); focal cerebral I/R group (group I/R); isoflurane preconditioning group (group Ⅰ).Focal cerebral I/R was produced by mid-cerebral artery occlusion in anesthetized rats.The rats inhaled 2 h of 2％ isoflurane and focal cerebral I/R was produced 24 h later in group I.The neurological deficits were scored at 24 h of reperfusion.The animals were then sacrificed.The brains were immediately removed for determination of the infarct size.The expression of 5-LOX,myeloid differentiation factor88 (MyD88) and nuclear factor kappa B (NF-κB) protein and mRNA was detected using Western blot and RT-PCR respectively.Results Compared with group S,the neurological deficit score was significantly increased,the infarct size was enlarged in groups I/R and I,the expression of 5-LOX,MyD88 and NF-κB protein and mRNA was up-regulated in group I/R,and the expression of 5-LOX mRNA and MyD88 protein and mRNA was up-regulated in group I (P ＜ 0.05).Compared with group I/R,the neurological deficit score was significantly lower,the infarct size was smaller,and the expression of 5-LOX,MyD88 and NF-κB protein and mRNA was lower in group I (P ＜ 0.05).Conclusion Isoflurane preconditioning can reduce focal cerebral I/R injury by down-regulating the expression of 5-LOX and inhibiting MyD88/NF-κB signaling pathway in rats.

Effect of isoflurane preconditioning on TLR4-MyD88 signaling pathway in ischemic penumbra following focal cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To investigate the effect of isoflurane preconditioning on Toll-like receptor 4 (TLR4)-myeloid differentiation factor 88 (MyD88) signaling pathway in ischemic penumbra following focal cerebral ischemia-reperfusion (I/R) in rats.Methods Fifty-four healthy male SD rats,aged 3 months,weighing 250-280 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),I/R group and isoflurane preconditioning group (group IP).Focal cerebral I/R was induced by middle cerebral artery occlusion.In groups I/R and IP,a nylon thread with rounded tip was inserted into the right internal jugular vein and threaded cranially until resistance was met.The middle cerebral artery was occluded for 2 h,followed by 24 h reperfusion.In group IP,the animals inhaled 2.0％ isoflurane for 2 h,and middle cerebral artery occlusion was performed at 24 h after the end of preconditioning.Neurological deficit was scored at 24 h of reperfusion and then the rats were sacrificed.Five rats in each group were chosen and the brains removed for measurement of the cerebral infarct volume.The right cerebral ischemic penumbra was removed for detection of the expression of HSP60,TLR4,MyD88 protein and mRNA by Western blot analysis and real time-PCR.Apoptosis was detected in the ischemic penumbra in the left 3 rats in each group using TUNEL.Apoptosis index (AI) was calculated.Results Neurological deficit scores and AI were significantly increased,the cerebral infarct volume was significantly enlarged,and the expression of HSP60,TLR4,MyD88 protein and mRNA was up-regulated in groups I/R and IP as compared with group S ( P ＜ 0.05).Isoflurane preconditioning significantly reduced the cerebral infarct volume and decreased neurological deficit scores and AI,and down-regulated the expression of HSP60,TLR4,MyD88 protein and mRNA (P ＜ 0.05).Conclusion The mechanisn by which isoflurane preconditioning protects ischenic penumbra following focal cerebral I/R may be related to inhibition of TLR4-MyD88 signaling pathway.