ABSTRACT
Objective:To determine the dose-effect relationship of oxycodone inhibiting responses to endotracheal intubation with combination of etomidate-rocuronium during induction of general anesthesia.Methods:A total of 120 patients, aged 20-63 yr, with body mass index of 18.0-25.2 kg/m 2, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective surgery under general anesthesia with tracheal intubation, were divided into 4 groups ( n=30 each) using a random number table method: group O 0.15, group O 0.23, group O 0.34 and group O 0.51.In O 0.15, O 0.23, O 0.34 and O 0.51 groups, oxycodone 0.15 mg/kg, 0.23 mg/kg, 0.34 mg/kg and 0.51 mg/kg were injected intravenously, respectively, 3 min later etomidate 0.3 mg/kg and rocuronium 0.8 mg/kg were intravenously injected in turn, and tracheal intubation was performed using Macintosh laryngoscope.Heart rate (HR), systolic blood pressure (SBP) and diastolic blood pressure immediately before intubation and the peak levels of HR, BP and diastolic blood pressure within 3 min after intubation were recorded.The response to endotracheal intubation was defined by the SBP and (or) HR having increased by more than 30% following intubation.The occurrence of adverse events during induction of anesthesia were recorded. Results:The rate of response to endotracheal intubation was 97%, 73%, 27%, and 3% in O 0.15, O 0.23, O 0.34 and O 0.51 groups, respectively.The median effective dose (ED 50) (95% confidence interval) of oxycodone inhibiting responses to endotracheal intubation was 0.259 (0.230-0.292) mg/kg, and the 95% effective dose (ED 95) (95% confidence interval) was 0.387 (0.358-0.420) mg/kg.Only the incidence of hypotension (27%) was significantly higher in group O 0.51 than in the other 3 groups ( P<0.05). Conclusion:With combination with etomidate-rocuronium, the ED 50 and ED 95 of oxycodone inhibiting responses to endotracheal intubation performed using Macintosh laryngoscope during induction of general anesthesia are 0.259 mg/kg and 0.387 mg/kg, respectively, and the optimum dose 0.51 mg/kg is recommended.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
Objective To evaluate the effect of resveratrol on the cognitive function after isoflurane anesthesia in obese rats.Methods Sixty SPF healthy male Sprague-Dawley rats,aged 4 weeks,weighing 70-80 g,were divided into 4 groups (n=15 each) using a random number table:normal diet plus 30％ O2 group (group N+O),high-fat diet plus 30％ O2 group (group H+O),high-fat diet plus 2％ isoflurane group (group H+I),and reveratrol plus high-fat diet plus 2％ isoflurane group (group R+H+I).Rats were fed a normal diet for 8 weeks in N+O group,while animals were fed a high-fat diet for 8 weeks in H+O,H+I,and R+H+I groups.Starting from 9th week,reveratrol 40 mg · kg-1 · d-1 was given into the stomach through a gastric tube for 7 consecutive days in group R+H+I,while the equal volume of 1％ carboxymethyl cellulose sodium was given instead of reveratrol in the other groups.After the end of drug intervention,animals were exposed to the mixture of 70％ nitrogen and 30％ oxygen for 4 h in N+O and H+O groups or to 2％ isoflurane for 4 h in H+I and R+H+I groups.Ten rats were randomly selected on 2nd day after inhaling isoflurane,and Morris water maze test was performed to assess the cognitive function.Five rats were randomly sacrificed on 3rd day after inhaling isoflurane,brains were removed,and hippocampi were isolated for determination of the expression of brain-derived neurotrophic factor (BDNF),tyrosine kinase B receptor (TrkB) and phosphorylated TrkB (p-TrkB) in hippocampal tissues by Western blot.The p-TrkB/TrkB ratio was calculated.Results Compared with group N+O,the escape latency was significantly prolonged on 3rd and 4th days,the exploration time spent on the original target quadrant was shortened,the expression of BDNF and p-TrkB in hippocampal tissues was down-regulated,and the p-TrkB/TrkB ratio was decreased in group-H+O (P＜0.05).Compared with group H+O,the escape latency was significantly prolonged on 2nd-5th days,the exploration time spent on the original target quadrant was shortened,the expression of BDNF and p-TrkB in hippocampal tissues was down-regulated,and the p-TrkB/TrkB ratio was decreased in group H+I (P＜0.05).Compared with group H+I,the escape latency was significantly shortened on 2nd-5th days,the exploration time spent on the original target quadrant was prolonged,the expression of BDNF and p-TrkB in hippocampal tissues was up-regulated,and the p-TrkB/TrkB ratio was increased in group R+H+I (P＜0.05).Conclusion Resveratrol can mitigates the cognitive dysfunction after isoflurane anesthesia in obese rats,and the mechanism may be related to promoting the activation of BDNF/TrKB signaling pathway.
ABSTRACT
Objective To evaluate the effect of high-level spinal cord injury(SCI)on the expression of mitochondrial voltage-dependent anion channel 2(VDAC2)in rat cardiomyocytes.Methods Forty-eight pathogen-free healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 2 groups(n=24 each)using a random number table:sham operation group(group S)and high-level SCI group(group H).The animals were anesthetized with intraperitoneal chloral hydrate and subjected to SCI using the modified Allen weight-drop method in group H.The spinal cord was only exposed in group S.At 6,12,24 and 48 h after SCI(T1-4),6 rats in each group were randomly selected and sacrificed,and myocardial specimens were collected from the cardiac apex for microscopic examination of the cell morphology(with a transmission electron microscope) and for determination of cell apoptosis(by TUNEL assay),expression of Bax,Bcl-2 and VDAC2 protein and mRNA in cardiomyocytes(by Western blot and real-time polymerase chain reaction,respectively).The apoptosis rate and ratios of Bax/Bcl-2 protein and mRNA were calculated.Results Compared with group S,the apoptosis rate and ratios of Bax/Bcl-2 protein and mRNA were significantly increased at T1-4,the expression of VDAC2 protein and mRNA was significantly down-regulated at T2-4(P<0.05 or 0.01),and the pathologic changes of cardiomyocytes were aggravated in group H.Conclusion The mechanism of myocardial damage is related to down-regulation of mitochondrial VDAC2 expression in cardiomyocytes and promotion of cell apoptosis in rats with high-level SCI.
ABSTRACT
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.
ABSTRACT
Objective To investigate the effects of different ventilation modes on PaCO 2 and PaO 2 during laparoscopic surgery in the Trendelenburg position.Methods Forty cases of laparoscopic resection of rectal cancer were randomly divided into group A (n =20)and group B (n =20).Under general anesthesia,all the cases were in volume-controlled ventilation mode (VCV,tidal volume=10 ml/kg)before pneumoperitoneum.After ventilating in VCV mode for 20 minutes with a lower tidal volume (8 ml/kg),group A was converted to PCV mode for 20 minutes and then back to the VCV mode for 20 minutes again.Next we switched to PCV combined with PEEP (5 cm H 2 O)mode (PCV+PEEP)for 20 minutes.In group B,we only alternated PCV with PCV combined with PEEP.Arte-rial blood-gas analysis was obtained at each time when ventilating mode changed.Results In both group A and B,PaO 2 in VCV mode was less than that in PCV mode and PCV+PEEP mode (P <0.05),PaO 2 in PCV mode was also less than that in mode PCV+PEEP mode (P <0.05).PaCO 2 in PCV and PCV combined with PEEP mode was less than that in VCV mode (P <0.05 ),and there was no difference of PaCO 2 in PCV mode and PCV+PEEP mode.The pH value in VCV mode was less than that in PCV mode and PCV+PEEP mode (P < 0.05).There was no difference in pH value between PCV mode and PCV+PEEP mode.Conclusion PCV combined PEEP mode is beneficial ei-ther in increasing or decreasing of PaCO 2 during laparoscopic surgery in the Trendelenburg position comparing with single VCV mode or PCV mode.
ABSTRACT
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.
ABSTRACT
Objective To compare the accuracy of Marsh model and Schnider model for propofol target?controlled infusion ( TCI) system. Methods Eighty patients, aged 20-60 yr, of American Society of Anesthesiologists physical status ⅠorⅡ, with body mass index of 17?5-28?0 kg∕m2 , scheduled for e?lective gynecological operation under general anesthesia, were equally and randomly divided into either Marsh model group ( group M) or Schnider model group ( group S) using a random number table. The target plasma concentration was set at 3 μg∕ml in both groups. During TCI and at different time points after the end of TCI, the blood samples were collected for determination of blood propofol concentrations by high per?formance liquid chromatography with fluorescence detector. The difference between measured and predicted concentrations (△C) at each time point was calculated. The median performance error ( MDPE) , median absolute performance error ( MDAPE) , and wobble of propofol TCI system were calculated in each group. Results In M and S groups, the MDPE was 9. 90% and 14?00%, respectively; the MDAPE was 11?43% and 14?49%, respectively;the wobble was 7?77% and 7?79%, respectively. There was no sig?nificant difference in △C at each time point during TCI between group M and group S (P>0?05). After TCI was stopped, △C at each time point was significantly lower in group M than in group S ( P<0?05) . Conclusion Marsh model provides higher accuracy than Schnider model for propofol TCI system in the pa?tients undergoing gynecological operation.
ABSTRACT
Objective To evaluate the effect of high-level spinal cord injury (SCI) on the myocardial energy metabolism in rats.Methods Sixty healthy male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n=30 each) using a random number table:sham operation (group S) and SCI group.SCI was induced in anesthetized rats by dropping a 10 g weight onto C7 spinal cord from 5 cm height falling freely inside a vertical hollow glass tube.At 6,12,24,48 and 72 h after SCI,6 rats in each group were chosen and arterial blood samples were taken for measurement of serum creatine kinase (CK) and creatine kinase isoenzyme-MB (CK-MB) activities.The rats were then sacrificed and myocardial specimens were obtained for examination of myocardial ultrastructure and for determination of ATP weight ratio,levels of Na+-K+-ATPase,Ca2+-Mg2+-ATPase,non-esterified fatty acids (NEFA) and lactic acid (LD),and expression of peroxisome proliferator-activated receptor alpha (PPARα) mRNA and protein (using fluorescent quantitative PCR and Western blot).Results Compared with group S,the serum CK and CK-MB activities were significantly increased,the ATP weight ratio,activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase and levels of NEFA and LD were decreased,and the expression of PPAR-α mRNA and protein was down-regulated in SCI group.No pathological changes of myocardium were found in group S,and the pathological changes of myocardium were obvious in SCI group.Conclusion High-level SCI can lead to decrease in the myocardial energy metabolism in rats,and down-regulated expression of PPARα is involved in the mechanism.
ABSTRACT
Objective To evaluate the effect of isoflurane preconditioning on cell apoptosis during renal ischemia-reperfusion (I/R) in rats.Methods Thirty male Sprague-Dawley rats, aged 12-14 weeks, weighing 300-320 g, were randomly divided into 3 groups (n =10 each) using a random number table: sham operation group (group S);I/R group;isoflurane preconditioning plus I/R group (group Iso+I/R).The rats were anesthetized with intraperitoneal pentobarbital sodium 30 mg/kg.To establish the model of renal I/R injury, the right kidney was removed, and the left renal pedicle was occluded for 30 min with atraumatic mini-clamp for 30 min, followed by 2 h reperfusion.In Iso+I/R group, 1.5％ isoflurane was inhaled for 1 h, followed by 30 min of washout before I/R.In S and I/R groups only oxygen 2 L/min was inhaled for 1 h.Arterial blood samples were taken at 2 h of reperfusion to determine the concentrations of serum creatinine (Cr), blood urea nitrogen (BUN) and cystatin C (CysC).The animals were then sacrificed, and left kidneys were sampled for determination of the cell apoptosis (by TUNEL), expression of p53, Bax and Bcl-2 mRNA (using real-time fluorescent quantitative PCR), and expression of Bax, Bcl-2, and caspase-3 (by Western blot analysis).The Bcl-2/Bax ratio was calculated.Results Compared with group S, the serum Cr, BUN, and CysC concentrations were significantly increased, the Bcl-2/Bax ratio was decreased, Bcl-2 mRNA, Bax mRNA, Bcl-2, Bax and caspase-3 expression was up-regulated, and AI was increased in group I/R.Compared with group I/R, the serum Cr, BUN, and CysC concentrations were significantly decreased, the Bcl-2/Bax ratio was increased, Bcl-2 mRNA, Bax mRNA, Bax and caspase-3 expression was down-regulated, and AI was decreased in group Iso+I/R.There was no significant difference in p53 mRNA expression among the three groups.Conclusion Regulating the balance between Bcl-2 and Bax and inhibiting apoptosis in kidney cells are involved in the mechanism by which isoflurane preconditioning reduces renal I/R injury in rats.
ABSTRACT
Objective To investigate the effect of creatine phosphate sodium on myocardial protection and calcium-sensitive receptor (CaSR) expression following high-level spinal cord injury.Methods Thirty healthy male SD rats weighing 250-300 g were assigned to sham operation,12-hour injury,24-hour injury,12-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium,and 24-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium according to the random number table,with 6 rats in each group.High-level spinal cord injury was induced at C7 segment by dropping a 10 g weight falling freely along the hollow glass tube from a 5 cm height.Level of blood troponin Ⅰ (cTnⅠ) was measured.Myocardial tissues were collected to study ultrastructure of myocardial cells under transmission electron microscope and CaSR expression using fluorescence quantitative PCR and Western blotting.Results cTnⅠ level was (0.031 ±0.002) U/L and (0.026 ± 0.001) U/L in 12-and 24-hour injury groups,but it was reduced to (0.023 ± 0.002) U/L and (0.018 ± 0.006) U/L at the same time point in treatment groups (P ＜ 0.05).Whereas either in injnry or treatment groups,cTnⅠ level was higher than (0.004 ± 0.002) U/L in sham operation group (P ＜ 0.05).CaSR mRNA level was (0.991 ±0.146) × 10-3 and (1.245 ±0.204) × 10-3 in 12-and 24-hour injury gronp and decreased to (0.880 ± 0.096) × 10-3 and (0.782 ± 0.138) × 10 3 at the same time point in treatment groups (P ＜ 0.05),but all were higher than (0.437 ± 0.065) × 10-3 in sham operation group (P ＜ 0.05).CaSR protein expressed in 12-and 24-hour injury group was (0.627 ±0.066) × 10 3 and (0.809 ±0.154) ×10 3 and lowered to (0.505 ±0.176) × 10-3 and (0.524 ±0.138) × 10-3 at the same time point in treatment groups,but all were higher than (0.331 ± 0.102) × 10-3 in sham operation group (P ＜ 0.05).Transmission electron microscopy demonstrated normal myocardial ultrastructure in sham operation group but impairment in injury groups,but the impairment was significantly improved in treatment groups.Conclusion Creatine phosphate sodium can decrease cTnⅠ level,attenuate the damage to myocardial ultrastructure and down-regulate CaSR after high-level spinal cord injury.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
Objective To investigate the effects of exogenous pulmonary surfactant(PS)on the inflammatory response in rats with ventilator-induced lung injury(VILI).Methods Twenty-eight adult male Wistar rats weighing 310-388 g were randomly divided into 4 groups(n =7 each):normal control group(group C),group VILI,group PS and air control group(group A).VILI was produced by high-pressure ventilation(HPV)with peak inspiratory pressure(PIP)40 cm H2 O,respiratory rate(RR)20 hpm and without positive end-expiratory pressure(PEEP)for 20 min.The rats were sacrificed by exsanguination immediately after anesthesia and after VILI in groups C and VILI,respectively.In groups PS and A,PS 100 mg/kg(50 mg/ml)and the equal volume of air were injected into the trachea via the airway after the tracheal edema fluid was removed respectively,and the rats were mechanically ventilated(Vr 10 ml/kg,RR 45 bpm and PEEP 7.5 cmH2O)for 120 min and then sacrificed by exsanguination.The blood samples were taken from femoral artery for determination of the plasma levels of IL-6,IL-10,MIP-2 and TNF-α by ELISA.The tracheal edema fluid was collected to determine the protein concentration by Bradford method.The lungs were removed for microscopic examination and the number of neutrophils was counted under microscope.Results There was no significant difference in the plasma levels of TNF-α among the four groups(P ＞ 0.05).Compared to group C,the plasma levels of MIP-2,IL-10 and IL-6 were significantly increased and the number of neutrophils was significantly enlarged in group VILI(P ＜ 0.05).The number of neutrophils was significantly smaller in group PS than in group A(P ＜ 0.05).There was no significant difference in the plasma levels of MIP-2,IL-10 and IL-6 between groups PS and A(P ＞ 0.05).The inflammatory damage in lung tissues was observed obviously in groups VILI,A and PS.Conclusion When exogenous PS is used to treat the rats with VILI,PS reduces neutrophil recruitment,but can not inhibit the release of inflammatory cytokines.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
ObjectiveTo investigate the role of myocardial calcium-sensing receptor (CaSR) in a rat model of high-level spinal cord injury (SCI).MethodsEighteen healthy male SD rats weighing 250-300 g were randomly divided into 2 groups:sham operation group(group S,n ＝6) and SCI group(n ＝ 12).SCI model was induced by dropping a 10 g weight onto spinal cord (C7) in freely vertical falling along the hollow glass tube from 5 cm height.The blood samples were taken 12 and 24 h after SCI in group SCI and 12 h after SCI in group S,and serum activity of creatine kinase(CK) and MB isoenzyme of creatine kinsse(CK-MB) were measured.Then myocardium specimens were obtained for uhrastructure examination and determination of CaSR mRNA and protien expression by fluorescence quantitative RCR and Western blot.Results Serum activities of CK and CK-MB and CaSR mRNA and protein expression were higher in group SCI than in group S.Serum activity of CK and CaSR mRNA expression were higher,and serum activity of CK-MB was lower at 24 h after SCI than that at 12 h after SCI.There was no significantly difference in CaSR protein expression between the two time points in group SCI.The ultrastructure examination showed that myocardial injury was found in group SCI.ConclusionThe expression of CaSR is up-regulated after SCI in rats,which might be the mechanism of myocardial injury after SCI.
ABSTRACT
Objective To investigate the effects of sevoflurane preconditioning on renal ischemia-reperfusion(I/R)injury in rats.Methods Twenty-four adult male SD rats weighing 250-300 g were randomly divided into 3 groups(n=8 each):sham operation group (group S);I/R group; sevoflurane preconditioning group (group SP). After the rats underwent right nephrectomy, renal I/R was produced by occlusion of left renal artery for 45 min followed by reperfusion in I/R and SP groups.In group SP, the rats inhaled 2.2% sevoflurane for 1 h, then the inhalation was stopped and renal ischemia was performed 10 min later. Venous blood samples were collected at 2 h of reperfusion to determine the concentrations of serum creatinine(Cr), urea nitrogen (BUN), cystatin C (Cys C) . The renal tissues were obtained for microscopic examination, and Paller's score was recorded. Results Compared with group S, there was no significant difference in the serum Cr and BUN concentrations (P＞0.05), while the serum Cys C concentration and Paller's score for acute renal tubular injury were significantly increased in group I/R(P＜0.05). The serum Cys C concentration and Paller's score were significantly lower in group SP than in group I/R(P＜0.05).I/R-induced renal injury was significantly reduced in group SP compared with group I/R. Conclusion Preconditioning with sevoflurane can provide significant protection against renal I/R injury.