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Objective To evaluate the effect of resveratrol on mitochondrial function in renal tubular epithelial cells of rats with sepsis-induced acute kidney injury.Methods Ninety-six healthy SpragueDawley rats of both sexes,aged 5-7 weeks,weighing 180-220 g,were divided into 4 groups (n =24 each) using a random number table method:sham operation group (Sham group),sepsis group (group Sep),sepsis plus vehicle group (Sep+Ⅴ group) and sepsis plus resveratrol group (Sep+R group).Sepsis was induced by cecal ligation and puncture (CLP).Normal saline 0.5 ml,vehicle 0.5 ml and resveratrol 10 mg/kg were intraperitoneally injected at 6,12 and 18 h after CLP in Sep,Sep+Ⅴ and Sep+R groups,respectively.At 24 h after CLP,serum concentrations of creatinine (Cr) and blood urea nitrogen (BUN) were measured,and kidney tissues were obtained for examination of the pathological changes (using transmission electron microscopy),and the damage to the renal tubules was scored.The renal tubular epithelial cells (RTECs) were isolated from the kidney cortex at 24 h after CLP for determination of mitochondrial transmembrane potential (by flow cytometry with the fluorescent probe JC-1),intracellular ATP content,mitochondrial permeability transition pore (mPTP) opening,lipid peroxide (LPO) content,and lysosomal membrane permeability.Results Compared with group Sham,the serum concentrations of Cr and BUN,renal tubular damage score,mPTP opening,LPO content and lysosomal membrane permeability were significantly increased,and mitochondrial transmembrane potential and ATP content were decreased in Sep and Sep+Ⅴ groups (P＜0.01).Compared with Sep and Sep+Ⅴ groups,the serum concentrations of Cr and BUN,renal tubular damage score,mPTP opening,LPO content and lysosomal membrane permeability were significantly decreased,and mitochondrial transmembrane potential and ATP content were increased in group Sep+R (P＜0.05).Conclusion Resveratrol improves mitochondrial function in renal tubular epithelial cells of rats with sepsis and reduces acute kidney injury,and the mechanism may be related to inhibiting oxidative stress and decreasing the lysosomal membrane permeability.
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Objective To determine if polydatin inhibited oxidative stress and inflammatory response in rats with sepsis-induced acute kidney injury (AKI).Methods Seventy-two rats (weighing 180-220 g) were randomly divided into the following groups: sham group, cecal ligation and puncture (CLP) CLP+normal saline group (group CN), group CLP+vehicle (group CV), and group CLP+polydatin (group CD) (n=18 each).Rats in groups CN, CV and CD underwent CLP to mimic sepsis-induced AKI.In sham group, the cecum was not ligated or punched, and the remaining procedures were the same as in group CLP.Normal saline, vehicle, and 30 mg/kg polydatin were administered at 6, 12, and 18 hours after CLP via the tail vein.At 24 hour post CLP, two clinically used markers of AKI, blood urea nitrogen (BUN), and creatinine (Cr) were tested, pathological changes of kidney tissue was observed under light microscopy in each group.Renal tubular damage assessment was carried out.Malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) content of renal tissue, serum cytokines such as TNF-α, IL-1β and IL-6, were also measured in each group at 24 hours after CLP.Results Compared with sham group, multiple indexes such as BUN, Cr, tubular injury scores, MDA content of renal tissue, and serum cytokines incluing TNF-α, IL-1β and IL-6 increased significantly (P<0.01), while SOD and GSH levels of renal tissue significantly decreased in groups CN and CV (P<0.01).Compared with groups CN and CV, the indicators such as BUN, Cr, tubular injury scores, MDA content of renal tissue, and serum cytokines such as IL-1β and IL-6 significantly decreased (P<0.05);while SOD and GSH levels of renal tissue significantly increased (P<0.05).Conclusion Sepsis caused by sepsis cecal ligation and puncture can cause acute kidney injury.Polydatin could alleviate kidney damage by attenuating systemic inflammatory response and inhibiting oxidative stress of renal tissue.
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Objective To evaluate the efficacy of an airway topical anesthesia catheter for topical anesthesia using a spray-as-you-go technique via the fiberoptic bronchoscope (FOB).Methods Forty American Society of Anesthesiologists physical status Ⅰ-Ⅲ patients with obstructive sleep apnea syndrome,aged 20-64 yr,with body mass index of 23-35 kg/m2,with no upper respiratory tract infection within 1 week before operation,scheduled for elective uvulopalatopharyngoplasty,were divided into 2 groups (n =20 each) using a random number table:routine control group (group C) and FOB-airway topical anesthesia catheter group (group F).In group C,the pharynx and larynx were sprayed with lidocaine FOB by using a laryngo-tracheal mucosal atomization device,and cricothyroid membrane puncture was performed and then lidocaine was injected.In group F,airway topical anesthesia was performed using a spray-as-you-go technique via the FOB with an airway topical anesthesia catheter spraying lidocaine via the nose.At 5 min after topical anesthesia of the airway,FOB-guided intubation was performed,and dexmedetomidine was intravenously infused at 0.1 μg · kg-1 · min-1 for sedation in both groups.Ramsay sedation scores were assessed after topical anesthesia and before intubation.The scores for the intubating condition and tolerance of tracheal tube were assessed during FOB-guided intubation.Successful intubation and the development of responses to intubation and hypoxemia were recorded.The patients were followed up one day after the end of operation,and parents' satisfaction with the procedure of intubation was recorded.Results Compared with group C,the intubating condition score,tolerance of tracheal tube score,success rate of intubation at first attempt and rate of parents' satisfaction with the procedure of intubation were significantly increased,and the incidence of responses to intubation was decreased (P＜0.05),and no significant change was found in Ramsay sedation scores before intubation and incidence of hyoxemia in group F (P＞0.05).Conclusion When the FOB is used to guide awake nasotracheal intubation,the airway topical anesthesia catheter provides better efficacy,better intubating conditions,and fewer side effects when applied for topical anesthesia using a spray-as-you-go technique via the FOB,it can be easily accepted by the patients and the efficacy is better that of routine airway topical anesthesia.
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Objective To compare the development of postoperative cognitive dysfunction ( POCD) under total inhalation anesthesia with sevoflurane versus total intravenous anesthesia with propofol in elderly patients. Methods Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients, aged 65-77 yr, weighing 43-78 kg, with preoperative Mini?Mental State Examination ( MMSE) score≥25, scheduled for elective surgery for oral and maxillofacial carcinoma, were divided into 2 groups ( n=30 each) using a random number table: total inhalation anesthesia with sevoflurane group ( group S) and total intravenous anesthesia with propofol?based anesthesia group ( group P ) . The patients were tracheally intuba?ted under local infiltration anesthesia. In group S, anesthesia was induced with inhalation of 8% sevoflurane (oxygen flow rate 8 L∕min), rocuronium 0?9 mg∕kg was injected intravenously when the bispectral index ( BIS) value reached 45, and the patients were mechanically ventilated; anesthesia was maintained with inhalation of sevoflurane with the end?tidal concentration of 2%-3%. In group P , anesthesia was induced with iv propofol 2 mg∕kg and sufentanil 0?3μg∕kg, rocuronium 0?9 mg∕kg was injected intravenously when the BIS value reached 45, and the patients were mechanically ventilated; anesthesia was maintained with target?controlled infusion of propofol ( target plasma concentration 3-5μg∕ml) and remifentanil ( target plas?ma concentration 3-5 ng∕ml). In both groups, intermittent iv boluses of cisatracurium 0?04 mg∕kg were given to maintain muscle relaxation during operation, and BIS value was maintained at 40-60 during opera?tion. Before intubation ( T1 ) , immediately after onset of intubation ( T2 ) , at 10 min of intubation ( T3 ) , immediately after begninning of skin incision ( T4 ) , while operating on the base of tongue or sawing the low?er jaw ( T5 ) , at the end of operation ( T6 ) and on the morning of the postoperative day 1 ( T7 ) , blood sam?ples from the elbow vein were collected for determination of plasma norepinephrine and epinephrine concen?trations by high?performance liquid chromatography and electrochemistry. At T1?7 and on the morning of the postoperative day 3 ( T8 ) , blood samples from the elbow vein were collected for measurement of plasma cor?tisol concentrations by radioimmunoassay. MMSE was used to assess the cognitive function on the postopera?tive day 7. MMSE score0?05) . Conclusion Although the probability of the development of POCD is low when the two anesthetic methods are used, total intravenous anesthesia with propofol?based anesthesia induces a marked decrease in periop?erative stress responses when compared with total inhalation anesthesia with sevoflurane.
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Objective To investigate the effects of drag-reducing polymers on microcirculation in 40%total body surface area burn-injured rats. Methods SD rats were randomized into control group, drag-reducing polymer (DRPs) group and normal saline (NS) group (5 minutes after scald, drag-reducing polymer or saline was injected for fluid resuscitation). Wet dry weigh ratio of lung, histopathologic changes and arterial blood gas at 24 hour were respectively measured by wet dry weigh ratio method, hematoxylin-eosin (HE) staining and arterial blood gas analysis. The velocity of flow of red cell in oblique ridge and the survival time of burn-injured rats were observed. Results Compared with control group, rats in NS group exhibit significant lung injury characterized by a high W/D (P < 0.01), accumulation of a large number of neutrophils in HE stain, low partial pressure of oxygen (PO2) and high lactate (Lac) (P<0.05 or P<0.01) in arterial blood. Compared with the NS group, DRPs treatment rats exhibit significantly reduced lung injury characterized by W/D reducing (P < 0.05), the reduction of neutrophil infiltration, increased PO2, decreased Lac (P<0.05, P<0.01). In addition, DRPs treatment obviously increases the burn-induced low velocity of flow of red cell in oblique ridge (P<0.01). Moreover, the survival time of burned rats can be improved by DRPs treatment (P < 0.05). Conclusion DRPs ameliorates burn-induced acute lung injury, the mechanism may be through improving the burn-induced microcirculation disorders.
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Objective To evaluate the efficacy of T-joint endoscopy mask for fiberoptic bronchoscopy (FOB)-guided awake nasotracheal intubation in patients with cervical spinal cord injury.Methods Forty patients of both sexes aged 21-64 yr with fracture of cervical spine complicated by spinal cord injury scheduled for anterior decompression and interbody fusion under general anesthesia were randomly divided into 2 groups according to the technique for awake nasotracheal intubation (n =20 each):group nasal catheter and group T-joint endoscopy mask.Topical anesthesia of nasal cavity,pharynx,larynx and trachea with 2％ lidocaine was conducted and then remifentanil was continuously infused at 0.05-0.15 μg· kg-1 · min-1 in both groups.The incidence of hypoxemia and intubation time were recorded.Arterial blood samples were obtained for determination of PaO2 and PaCO2 before topical anesthesia (baseline),immediately before and 1 min after placement of FOB and immediately after nasotracheal intubation was accomplished.Results The incidence of hypoxemia was significantly lower in group Tjoint endoscopy mask (0) than in group nasal catheter (25％) (P ＜ 0.05).The PaO2 during nasotracheal intubation was significantly higher in group T-joint endoscopy mask than in group nasal catheter (P ＜ 0.05).There was no significant difference in PaCO2 and intubation time between the 2 groups (P ＞ 0.05).Conclusion T-joint endoscopy mask facilitates awake nasotracheal intubation without affecting oxygen inhalation in patients with cervical spinal cord injuries.
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Objective To construct F344 rat bone marrow mesenchymal stem cell line (MSC) modified with human hepatocyte growth factor (hHGF) gene.Methods Recombinant virus containing hHGF was obtained by transfecting the packaging cell line 293 FT with lentiviral vector pLV/EF1α-hHGF-IRES-eGFP.MSCs derived from F344 rat bone marrow were then tranfected with packed lentiviral vector.Purified MSCs expressing hHGF was obtained by screening culture with G418.MSCs and MSCs transfected with empty vector were used as control.The expression of hHGF protein was detected by Western blot (eGFP-MSCs).The hHGF-transfected MSCs were cultured in osteoblast-inducing culture medium and osteoblast phenotype was assayed by alizarin Red staining.The cells were also cultured in adipogenesis medium and stained with Oil Red O for identification.Results The expression of hHGF protein was significantly up-regulated in the hHGF-MSCs as compared with MSCs and eGFP-MSCs.hHGF-MSCs readily differentiated into mineralizing cells or adipocytes when incubated in differentiation medium.Conclusion A F344 rat MSC line that stably expresses HGF is successfully established.
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Objective To investigate the effect of human hepatocyte growth factor (hHGF) genetic modification on the ameliorating effects of mesenchymal stem cells (MSCs) implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension (PH).Methods MSCs were obtained from F344 rats and transduced with lentiviral vector modified with human HGF (hHGF-MSCs) or empty vector (EGFP-MSCs).Sixty-six 7 week old male F344 rats weighing 180-250 g were used in this study.PH was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg injected at 2 weeks after operation.The animals with PH were randomly divided into 3 groups:control group (group C),EGFP-MSCs group (group E) and HGF-MSCs group (group H).Groups H and E received hHGF-MSCs or EGFP-MSCs 5 × 105 in DMEM 1 ml iv at 3 weeks after subcutaneous MCT injection,while group C received plain DMEM 1 ml.Mean pulmonary arterial pressure (mPAP) was measured and right ventricular hypertrophy and angiogenesis in the lung were assessed and the content of rat HGF (rHGF) and hHGF protein in lung tissue and pulmonary capillary density (by immuno-histochemistry) was measured at 2 weeks after MSCs implantation.The survival rates within 45 days after MCT administration were compared among the 3 groups.Results No hHGF was detected in groups C and E.Both hHGF-MSCs and EGFP-MSCs significantly reduced MPAP and right ventricular hypertrophy and increased pulmonary capillary density and survival rates in groups H and E as compared with group C and the efficacy of hHGF-MSCs was significantly greater than that of EGFP-MSCs.Barium angiography revealed that distal pulmonary vasculature was significantly increased in group H as compared with groups E and C.The survival of the rats receiving hHGF-MSCs was significantly longer in group H than that in groups E and C.Conclusion hHGF genetic modification can improve the ameliorating effects of MSCs implantation on PH-related microvascular rarefaction.
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Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) genetically modified with human hepatocyte growth factor gene (hHGF) on angiogenesis in the rat lung.Methods Twenty F344 rats,aged 2 months,weighing 200-250 g,were randomly divided into 2 groups ( n =10 each):HGF group and control group (group C).MSCs genetically modified with hHGF was injected through the external jugular vein in group HGF.While the equal volume of DMEM culture medium (1 ml) was given instead in group C.The mean pulmonary artery pressure was detected at 28 days after transplantation.Then the rats were sacrificed and the lungs were removed for determination of the content of hHGF,expression of proliferating cell nuclear antigen (to reflect the degree of endothelial cell proliferation showed by the small pulmonary vessels) and Ⅷ factor (to reflect the density of the small pulmonary vessels),and microscopic examination.Results Compared with group C,no significant change was found in mean pulmonary artery pressure ( P ＞ 0.05),while the content of hHGF,degree of endothelial cell proliferation,and density of the small pulmonary vessels were significantly increased in group HGF ( P ＜ 0.01).No change was found in the structure of the small pulmonary vessels in group HGF.Conclusion Transplantation of MSCs genetically modified with hHGF can promote angiogenesis in the rat lung.
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Objective To investgate the changes in the expression of hepatocyte growth factor (HGF)and c-met in the lungs in a rat model of pulmonary hypertension.Methods Eighty 7 week old male SD rats weighing 180-250 g were randomly divided into 2 groups ( n =40 each ):control group (group C) and pulmonary hypertension group (group PH).Pulmonary hypertension was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg 2 weeks later.Pulmonary artery pressure and the ratio between the weight of right ventricle and left ventricle + interventricular septum ( RV/LV + S) were measured at 7,14,21 and 28 d after MCT administration.HGF and c-met protein and mRNA expression and TGF-β content in the lung tissue were determined.Results Pulmonary hypertension and right ventricular hypertrophy associated with hypertrophy of pulmonary artery tunica media and muscularization of small pulmonary arteries developed after MCT administration in PH group.In PH group HGF protein and mRNA expression in the lungs was significantly down-regulated as compared with group C.There were no significant differences in c-met protein and mRNA expression in the lungs between the 2 groups.The TGF-β content in the lungs was significantly increased in group PH as compared with group C.Conclusion Decrease in HGF production in the lungs plays an important role in the pulmonary hypertension.Increasing of pulmonary TGF-β may play an important role in the down-regulation of pulmonary HGF expression during pulmonary hypertension.
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Objective To compare the efficacy of orotracheal intubation after rapid induction and nasotracheal intubation after slow induction assisted by fiberoptic bronchoscope (FOB) in patients with obstructive sleep apnea syndrome (OSAS).Methods Forty ASA Ⅱ patients with OSAS and hypertension, aged 38-64 yr, weighing 82-123 kg, undergoing uvulopalatopharyngoplnsty, were randomly allocated into 2 groups (n = 20 each): group Ⅰ awake nasotracheal intubation through FOB after slow induction and group Ⅱ orotracheal intubation through laryngoscope and FOB after rapid induction. MAP and HR were recorded immediately after anesthesia induction and intubation. The intubation time was recorded, and rate of successful intubation, incidences of tschycardia, hypertension and myocardial ischemia during intubation, and satisfactory rate of otolaryngologists with airway management were calculated. Results The rate of successful intubation were significantly higher, MAP and HR after induction were significantly lower, incidences of tachycardia, hypertension and myocardial ischemia during intubation were significantly lower, and satisfactory rate of otolaryngologists was significantly higher in group Ⅱ than in group Ⅰ ( P < 0.05). Four patients in group Ⅰ had nasal bleeding after extubation. Conclusion Orotracheal intubation after rapid induction assisted by FOB provides less hemodynamic response to endotracheal intubation and higher success rate with less injury to the airway in patients with OSAS than nasotracheal intubation after slow induction assisted by FOB.