ABSTRACT
Objective To explore the effect and mechanism of IncRNA EVADR on the proliferation and migration in human colorectal cancer cell lines HCT116 and LOVO. Methods HCT116 and LOVO cell lines were transfected with IncRNA EVADR by overexpressing lentivirus system. CCK8 assay was performed to measure the growth of HCT116 and LOVO cells after overexpression of EVADR. Transwell migration was performed to determine if EVADR promote HCT116 and LOVO cells migration. Finally, the expression of Ecadherin and transcription factor Snail, Slug, ZEB1 and ZEB2 were detected by Western blot and realtime quantitative PCR respectively. Results We successfully established colorectal cancer cells strains HCT116, LOVO which can stably overexpress IncRNA EVADR and the capacity of proliferation and migration in overexpression group was significantly improved (P＜0.05). The expression of Ecadherin was decreased while mesenchymal markers Snail, Slug, ZEB1 and ZEB2 were increased in EVADR overexpression HCT116 and LOVO cells. Conclusions Overexpression of IncRNA EVADR in HCT116 and LOVO cells can significantly promote the proliferation and migration of HCT116 and LOVO cells which may play an important role in regulating epithelial-mesenchymal transition in colorectal cancer cells.
ABSTRACT
Objective To investigate the effects of nicorandil on hypoxia-inducible factor (HIF)-1α mRNA and protein in lung tissue of one-lung ventilation.Methods Twenty-four clean New Zealand white rabbits were randomly divided into sham group (group S) (two-lung ventilation+thoracotomy),negative control group (group C) (one-lung ventilation + thoracotomy + saline),nicorandil group (group N) (one-lung ventilation+ thoracotomy+ nicorandil) and antagonist group (group J) (one-lung ventilation + thoracotomy + nicorandil + glibenclanide) equally.The implementation of mechanical ventilation depended on self-made double-lumen endotracheal tube after intravenous induction through ear marginal vein.Intravenous maintenance medicine was infused by trace injection pump after anesthesia induction.The implementation of thoracic surgery was simulated through one-lung and two-lung ventilation by auscultation,bubble test and direct observation.Group S was given anaesthesia only,no one-lung ventilation group S,the other three groups had single lung ventilation,and the drug was injected before the operation.Group N was infused nicorandil 100 ptg· kg-1 · h-1 before the implementation of single lung ventilation for 1 h.Group C was injected with the same amount of normal saline.Group J was intravenous infusion of glibenclamide 75 μg· kg-1 · h-1 and nieorandil 100μg · kg-1 · h-1 the implementation of single lung ventilation for 1 h.Then wet and dry weight ratio(W/D) and superoxide dismutase (SOD) activity were measured after non-ventilatory lung was processed and preserved.The expression of HIF-1α protein of non ventilatory lung tissue was detected by Western-blot in the four groups.The transcription of HIF-1α mRNA was detected by real-time quantitative real-time PCR (qRT-PCR) in all groups.Results W/D in groups C and J were significantly higher compared with that of groups S and N (P＜0.05).The activity of SOD in groups C and J was significantly lower compared with groups S and N (P＜0.05).The expression of HIF1α protein and transcription of HIF-1α mRNA in groups C,N and J were significantly higher than those in group S,and that of group N was significantly higher than those of groups C and J (P＜0.05).Conclsion Nicorandil has a protective effect on the collapse and inflation of non-ventilatory lung in rabbit with one-lung ventilation,reducing oxidative stress by SOD,acting on mito KATP and coming into play by up-regulation of HIF-1α.
ABSTRACT
[Objective]To radiolabel the PSMA aptamer A10-3.2 with 99mTc , and explore its biological characteristics in vivo and in vitro.[Methods]Using Succinimidyl 6-hydrazinonicotinate hydrochloride (SHNH) as the bifunctional chelating agent to label aptamer A10-3.2 with 99mTc, then tested for the stability in vitro, the specific uptake by prostate cancer LNCaP cells (PSMA+) , the characteristics of SPECT/CT imaging and biodistribution in LNCaP tumor-bearing NOD/SCID mice.[Results]The labeling rate and radiochemical purity of the products (99mTc-SHNH-A10-3.2) are(71.31 ± 6.78)% and 97.03%,respectively. 99mTc-SHNH-A10-3.2 had obvious target specificity for PSMA positive prostate cancer LNCaP cells, its uptake rate was significantly higher than PSMA nega?tive PC-3 cells (P<0.01). And in tumor-bearing mice, the tumor has a certain uptake and a high ratio of the tumor tissue to the mus?cle.[Conclusion]This study successfully constructed 99mTc-labeled PSMA-targeted aptamer A10-3.2, which has a good stability and targeting in vivo and in vitro, has a high tumor tissue/muscle ratio in tumor-bearing mice, which show that it may be a potential target?ed molecular imaging agent for prostate cancer.
ABSTRACT
Objective To investigate the effects and possible mechanisms of nicorandil on lung injury of the collapsed lung in one-lung ventilation.Methods Twenty-four clean Japanese big-ear rabbits were randomly divided into sham group (group S) (two-lung ventilation + thoracotomy),negative control group (group C) (one-lung ventilation + thoracotomy + saline),nicorandil group (group N) (one-lung ventilation+thoracotomy+nicorandil) and antagonist group (group J) (onelung ventilation+ thoracotomy+ nicorandil+ glyburide) equally.Mechanical ventilation was implemented through self-made double-lumen endotracheal tube after intravenous induction through ear marginal vein.Intravenous maintenance medicine was infused by trace injection pump after anesthesia induction.Thoracic surgery was simulated through one-lung or two-lung ventilation determined by auscultation,bubble test and direct observation.Then wet and dry weight ratio (W/D) and content of MDA were measured after non-ventilatory lung was processed and preserved.The expression of Akt,p-Akt and NF-κB protein in non-ventilatory lung tissue were detected by Western-blot in all groups.Results In respects of W/D and content of MDA,the other three groups had significant differences compared with group S (P ＜ 0.05).It was significantly lower in group N than in group C (P ＜0.05),and it was significantly higher in group J than in group N (P＜0.05).The expressions of pAkt protein and p-Akt/Akt in group N were significantly higher than those in group S and group C (P＜0.05).Those of group J were significantly lower than group N (P＜0.05).Compared with group S,the expression of NF-κB protein in group C was significantly higher (P＜0.05).That of group N was significantly lower than that of group C (P＜0.05).But that in group J was higher than that in group N (P ＜ 0.05).Conclusion Nicorandil has a protective effect on the collapse and inflation of non-ventilatory lung in rabbits under one-lung ventilation,acting on mitoKATP through PI3K/Akt,and down-regulating NF-κB to reduce IR-induced lung injury.
ABSTRACT
Objective To investigate the protective effects of dexmedetomidine on lung injury during one lung ventilation in patients undergoing lobectomia pulmonalis.Methods Sixty-four patients undergoing lobectomy in our hospital from May 2014 to February 2017 were selected.There were 38 males and 26 females,aged 42-75 years,ASA physical status Ⅱ or Ⅲ.Patients were divided into two groups according to different treatments,n=32 in each group.The patients in the observation group were given dexmedetomidine 0.5 μg· kg-1 · h-1 at 20 min before the induction of anesthesia,and adjusted to 0.2-0.5 μg·kg-1 ·h-1 after 10 min.The control group was given equal volume normal saline.The changes of polymorphonuclear neutrophil (PMN),myeloper oxidase (MPO),intra-pulmonary shunt rate(Qs/Qt),xanthine oxidase (XOD),vascular endothelial growth factor (VEGF) and nitric oxide (NO) concent ration were recorded at 10 min before induction (T0),beginning of OLV (T1),OLV for 60 min (T2),90 min (T3),postoperative 24 h (T4).Results Compared with T0,the PMN counts increased significantly at T2-T4 and the serum MPO and XOD concentrations were significantly increased (P＜0.05).The PMN counts,serum MPO and XOD concentration in the observation group were significantly lower than those in the control group (P＜0.05).Compared with T0,the serum VEGF concentration was significantly increased at T2 and T3,and the serum VEGF concentration in the observation group was significantly lower than that in control group at T3 (P＜0.05).The serum NO concentration at T2 and T3 in observation group was significantly higher than that in control group (P＜0.05).Conclusion Dexmedetomidine can reduce the inflammatory response of the lungs and has a protective effect on ischemia-reperfusion and injury in patients with single lung ventilation,and reduce the degree of oxidative stress,which plays a protective effect on lung.
ABSTRACT
Objective To investigate the effect of ultrasound-guided subcostal transverses abdominis plane (TAP) block with dexmedeto-midine after laparoscopic radical operation. Methods 40 patients underwent laparoscopic radical operation for colorectal cancer were ran-domized into dexmedetomidine group (group DEX) and control group (group CON). All the patients received ultrasound-guided subcostal TAP block after operation, Group DEX with dexmedetomidine 1μg/kg and 0.25%ropivacaine to 20 ml, and group CON with 0.25%ropiva-caine 20 ml. All the patients were assessed with Ramsay scores and the pain at rest and on coughing were assessed with Visual Analogue Scale (VAS), 2, 6, 12, 24 and 24 hours after operation. The highest level and the duration of sensory blockade, the first time and the total times of pressing the analgesia pump in the first day after operation, and the requirements of sufentanil were recorded. First flatus time, first diet time and the length of hospital stay were compared. Results The scores of VAS were significantly less (P<0.001), and the Ramsay scores were more in the group DEX than in the group CON (P<0.01) 2, 6 and 12 hours after operation; with the longer time of sensory blockade (P<0.001), the later to first press the analgesia pump (P<0.001), the less frequence of pressing the analgesia pump (P<0.001), and less dosage of sufentanil (P<0.001). The first flatus time, first diet time were significantly earlier in the group DEX than in the group CON (P<0.001), with the less length of total hospital stay (P<0.001). Conclusion Dexmedetomidine can promote the anaesthesia of ultra-sound-guided subcostal TAP block with ropivacaine and improve the recovery after laparoscopic radical operation.
ABSTRACT
Objective To evaluate the role of ERK1/2 signal transduction pathway in sevoflurane postconditioning-induced reduction of oxygen-glucose deprivation(OGD)injury in rat hippocampal slices.Methods Male adult SD rats weighing 80-100 g were anesthetized with ether and decapitated.The hippocampi were removed and sagittally sliced(400μm thick)and placed in artificial cerebrospinal fluid(aCSF)aerated with 95％ O2-5％CO2.Fifty hippoeampal slices were randomly divided into 5 groups(t =10 each):OGD group; 4％ sevoflurane postconditioning group(group Sevo); PD98059(specific inhibitor of ERK)group(group PD); dimethyl sulfoxide (DMSO)group; 4％ sevoflurane postconditioning + PD98059 group(group SPD).OGD was induced by incubating the slices in glucose-free aCSF aerated with 95％ N2-5％ CO2 for 15 min in group OGD.The hippocampal slices were perfused with aCSF saturated with 4％ sevoflurane for 30 min after OGD was induced in group Sevo.The hippocampal slices were perfused with aCSF containing PD98059 50 μmol/L for 10 min after OGD was induced in group PD.The hippocampal slices were perfused with aCSF containing DMSO 1 mol/L for 10 min after OGD was induced in group DMSO.The hippocampal slices were perfused with aCSF containing PD98059 50 μmol/L and aerated with 4％ sevoflurane for 30 min after OGD was induced in group SPD.The hippocampal slices were then perfused with plain aCSF for 1 h again in all the groups.The electrophysiological technique was used 1o record the amplitude of orthodromic population spike(OPS)in the stralum pyramidale of the CAI region.TTC staining was used to determine the degree of tissue injury.Results Compared with group OGD,the recovery amplitude and rate of OPS were significantly increased,and the degree of tissue injury was significantly decreased in group Sevo(P ＜0.01),while no significant change was found in each parameter in the other three groups(P ＞ 0.05).Compared with group Sevo,the recovery amplitude and tale of OPS were significandy decreased,and the degree of tissue injury was significantly increased in groups PD,DMSO and SPD(P ＜ 0.01).Conclusion ERK1/2 signal transduction pathway is involved in sevoflurane postcondilioning-induced reduction of OGD injury in tat hippocampal slices.
ABSTRACT
Objective To investigate the changes in the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) in the distal cerebrospinal fluid contacting neurons (CSF-CNs) in rats with neuropathic pain.Methods Forty-eight male adult SD rats aged 2-3 months weighing 230-270 g were randomly divided into 2 groups (n =24 each): sham operation group (group S) and chronic constrictive injury group (group CCI).Neuropathic pain was induced by CCI.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.Thermal withdrawal latency (TWL) was measured with radiation heat stimulus before (baseline) and 1,3,5,7,14 d after CCI in 4 animals at eachtime point in each group.The animals were then sacrificed.30％ cholera toxin subunit B with horse radish peroxidase 3 μl was injected into the lateral ventricle at 48 h before the animals were sacrificed at each time point.The brain tissue in the region of the midbrain aqueduct was taken.The number of distal CSF-CN expressing p-p38MAPK was calculated.p-p38MAPK was detected by histochemical staining.Results The TWL was significantly shortened at 1-14 d after CCI in group CCI as compared with group S.The number of distal CSF-CNs expressing p-p38MAPK was significantly increased at 5-14 d after CCI in group CCI as compared with group S.Conclusion The distal CSF-CNs may be involved in the neuropathic pain induced by CCI through p38MAPK signaling pathway.p38MAPK may contribute only to the maintenance but not development of neuropathic pain.
ABSTRACT
ObjectiveTo investigate the effects of diazoxide pretreatment on expression of phosphatidylinositol 3-kinase(PI3K) mRNA and protein serine/threonine kinase(Akt) mRNA in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation (H/R).MethodsThe SD rat myocardial microvascular endothelial cells were cultured.The cells were seeded in 96-well plates ( 100μd/hole) or in 6 cm diameter dishes (2 ml/dish) with the density of 1 × 106/ml and randomly divided into 4 groups ( n =25 each):normal control group (group C),H/R group,diazoxide pretreatment group (group DZ) and diazoxide pretreatment + 5-hydroxydecanoate (5-HD,a mitochondrial ATP-sensitive potassium channel blocker) group (group DZ + 5-HD).The cells were exposed to 2 h hypoxia followed by 2 h reoxygenation.Diazoxide 100 μmol/L and diazoxide 100 μmol/L + 5-HD 100 μmol/L were added to the culture medium 2 h before hypoxia in groups DZ and DZ + 5-HD respectively.The cell viability,apoptotic rate and expression of PI3K mRNA and Akt mRNA were detected at the end of reoxygenation.ResultsCompared with group C,the cell viability was significantly decreased,while the apoptotic rate increased and expression of PI3K mRNA and Akt mRNA up-regulated in group H/R (P ＜0.05 or 0.01).Compared with group H/R,the cell viability was significantly increased,while the apoptotic rate decreased and expression of PI3K mRNA and Akt mRNA up-regulated in group DZ (P ＜ 0.05 or 0.01).5-HD could inhibit diazoxide pretreatment-induced changes mentioned above (P ＜ 0.05 or 0.01 ).ConclusionDiazoxide pretreatment can reduce H/R injury by promoting PI3K gene and Akt gene transcription through activation of mitochondrial ATP-sensitive potassium channels in rat myocardial microvascular endothelial cells.
ABSTRACT
Objective To investigate the protective effects of isoflurane preconditioning plus mild hypothermia on isolated rat hearts against ischemia-reperfusion (I/R) injury. Methods Fifty male SD rats weighing 250-300 g were randomly divided into 5 groups ( n = 10 each): group Ⅰ control (group C); group Ⅱ I/R; group Ⅲ isoflurane preconditioning (group P); group Ⅳ mild hypothermia (group M) and group Ⅴ isoflurane preconditioning + mild hypothermia (group PM). The animals were anesthetized with intraperitoneal pentobarbital 40 mg/kg. The hearts were immediately removed and perfused with Krebs-Hensleit (K-H) solution aerated with 95% O2 and 5% CO2 at 10 kPa via aorta at 37℃ in a Langendorff apparatus. The hearts were made globally ischemic for 30 min followed by 60 min reperfusion in group Ⅱ-Ⅴ. In group P and PM the hearts were perfused with K-H solution saturated with 1.0% isoflurane for 15 min followed by 15 min washout before ischemia.In group M and PM the hearts were made ischemic at 31 ℃ and perfused at 37℃. LVEDP, LVSP, dp/dtmax,dp/dtmm and HR were measured after equilibration (baseline), immediately before ischemia, and at 30 and 60 min reperfusion. The infarct size and cytochrome C level in cytoplasm and mitochondria of myocytes were measured.Motochondrial ultrastructure was examined using electron microscope. Results Cardiac function was significantly better, the infarct size significantly smaller, the cytochrome C level in cytoplasm significantly lower, while the cytochrome C level in mitochondria of myocytes significantly higher in group Ⅲ-Ⅴ than in group Ⅱ and in group Ⅴ than in group Ⅲ. The cytochrome C level in cytoplasm was significantly lower, while the cytochrome C level in mitochondria of myocytes significantly higher in group Ⅴ than in group Ⅳ. Less damage to mitochondria was observed in group PM than in group I/R, P and M. Conclusion Isoflurane preconditioning combined with mild hypothermia provides better protection against myocardial I/R injury by attenuating the release of cytochrome C from mitochrondria.
ABSTRACT
Objective To investigate the effects of diazoxide pretreatment on the expression of NF-κB mRNA and fractalkine (FKN) mRNA in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation (H/R). Methods The SD rat myocardial microvascular endothelial cells were cultured. The cells were seeded in 96-well plates (100μl/hole) or in 6 cm diameter dishes (2 ml/dish) with the density of 1 × 106/ml and randomly divided into4 groups (n = 24 each): Ⅰ normal control group (group C), Ⅱ H/R group, Ⅲ diazoxide pretreatment group (group DZ), Ⅳ diazoxide pretreatment + mitochondrial ATP-sensitive potassium channel blocker 5-hydroxydecanoate (5-HD) group (group DZ + 5-HD). The cells were exposed to 2 h hypoxia followed by 2 h reoxygenation. Diazoxide 100 μmol/L and diazoxide 100 μmol/L + 5-HD 100μmol/L were added to the cultured medium 2 h before hypoxia in group DZ and DZ + 5-HD respectively. The cell vitality, apoptotic rate and expression of NF-κB mRNA and FKN mRNA were detected at end of reoxygenation. Results Compared with group C, the cell vitality was significantly decreased, apoptotic rate increased and the expression of NF-κB mRNA and FKN mRNA up-regulated in H/R group. Compared with group H/R, the cell vitality was significantly increased,apoptotic rate decreased and the expression of NF-κB mRNA and FKN mRNA down-regulated in group DZ. 5-HD could inhibit diazoxide pretreatment-induced changes mentioned above. Conclusion Diazoxide pretreatment can reduce H/R injury in rat myocardial microvascular endothelial cells through down-regulating the expression of NFκB and FKN, and the mechanism is related to activation of mitochondrial ATP-sensitive potassium channels.