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@#Objective To explore the relationship between preoperative fasting plasma glucose (FPG) and postoperative pulmonary complications (PPCs) in type 2 diabetic patients undergoing elective thoracoscopic lung resection, and provide a reference for prediction and prevention of PPCs in the clinic. Methods A retrospective analysis was performed on the type 2 diabetic patients who underwent elective thoracoscopic lung resection for the first time in our hospital from January 2017 to March 2021. According to the level of FPG one day before the operation, the patients were divided into three groups: a hypoglycemia group (<6.1 mmol/L), a medium level blood glucose group (≥6.1 mmol/L and <8.0 mmol/L) and a high blood glucose group (≥8.0 mmol/L). Besides, the patients were divided into a PPCs group and a non-PPCs group according to whether PPCs occurred. The risk factors for PPCs were analyzed by logistic regression analysis, and the predictive value of preoperative FPG level on PPCs was estimated by the area under the receiver operating characteristic curve (AUC). Results A total of 130 patients were included, including 75 (57.7%) males and 55 (42.3%) females with an average age of 63.5±9.0 years. Logistic regression analysis showed that compared to non-PPCs patients, the level of preoperative FPG (P=0.023) and smoking history ratio (P=0.036) were higher and the operation time was longer (P=0.004) in the PPCs patients. High FPG level on preoperative day 1 and longer operation time were associated with PPCs risk. Besides, the preoperative FPG of 6.79 mmol/L was the threshold value to predict the occurrence of PPCs [AUC=0.653, 95%CI (0.559, 0.747), P=0.003]. Conclusion There is a certain correlation between preoperative FPG level and postoperative PPCs, which may be used as an index to predict the occurrence of PPCs.
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Objective:To evaluate the relationship between Erbin and Bax/Bcl-xL-mediated cell apoptosis during sepsis-induced acute kidney injury in mice.Methods:Thirty-two SPF male wild type C57BL/6 mice, 32 SPF male Erbin (-/-) C57BL/6 mice, aged 6-8 weeks, weighing 20-30 g, were divided into 2 groups ( n=16 each) using the random number table method: wild type sham operation group (WT+ Sham group), wild type sepsis group (WT+ S group), Erbin(-/-) sham operation group (EKO+ Sham group), and Erbin(-/-) sepsis group (EKO+ S group). The sepsis model was established using the moderate cecal ligation and puncture (CLP) in anesthetized animals.The survival rates within 7 days after CLP were recorded.The serum concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), IL-1β, creatinine (Cr), blood urea nitrogen (BUN) and lactic dehydrogenase (LDH) were determined at 24 h after CLP.Then the renal tissues were taken for assessment of renal injury which was scored and for determination of the apoptosis rate (by TUNEL) and expression of cleaved-caspase-3, Bcl-xL and Bax (by Western blot). Results:Compared with sham operation groups, the survival rates were significantly decreased, the serum concentrations of IL-1β, IL-10, TNF-α, Cr, BUN and LDH, renal injury score and apoptosis rate were increased, the expression of Bax and cleaved-caspase-3 was up-regulated, and the expression of Bcl-xL was down-regulated in sepsis groups ( P<0.05). Compared with WT+ S group, the survival rates were significantly decreased, the serum concentrations of IL-1β, LDH, TNF-α, Cr and BUN and renal injury score were increased, the serum concentration of IL-10 was decreased, the apoptosis rate of renal tissues was increased, the expression of Bax and cleaved-caspase-3 was up-regulated, and the expression of Bcl-xL was down-regulated in EKO+ S group ( P<0.05). Conclusions:Erbin can inhibit Bax/Bcl-xL-mediated cell apoptosis and is involved in endogenous protective mechanism against sepsis-induced acute kidney injury in mice.
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Objective:To evaluate the role of ErbB2 interacting protein (Erbin) in sepsis-associated encephalopathy (SAE) in mice and the relationship with nod-like receptor thermoprotein domain associated protein 3 (NLRP3) inflammasomes.Methods:Sixty SPF-grade healthy male wild-type C57BL/6 mice and 60 Erbin (-/-)C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=30 each) by a random number table method: wild-type sham operation group (WT+ Sham group), wild-type SAE group (WT+ SAE group), Erbin (-/-) sham operation group (EKO+ Sham group) and Erbin (-/-) plus SAE group (EKO+ SAE group). The model of SAE was established by cecal ligation and perforation in anesthetized mice.Open field test (total distance moved) was performed at 7 days after establishing the model, new object recognition test (recognition index) was performed at 8 days after establishing the model, and Morris water maze test (time of staying at target quadrant) was performed at 10 days after establishing the model.The mice were sacrificed, and hippocampal tissues were removed for microscopic examination of pathologic changes (by HE staining) and for determination of neuron count, expression of NLRP3, caspase-1 and apoptosis-associated speck-like protein containing a CARD (ASC) (by Western blot), the number of NLRP3 positive cells (by immunohistochemistry), and contents of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-18 (by enzyme-linked immunosorbent assay). The cell survival rate was calculated. Results:Compared with group WT+ Sham, the time of staying at target quadrant was significantly shortened, the recognition index and cell survival rate were decreased, the contents of IL-1β, IL-18 and TNF-α and the number of NLRP3 positive cells were increased, and the expression of NLRP3, caspase-1 and ASC was up-regulated in group WT+ SAE ( P<0.05). Compared with group EKO+ Sham, the time of staying at target quadrant was significantly shortened, the recognition index and cell survival rate were decreased, the contents of IL-1β, IL-18 and TNF-α and the number of NLRP3 positive cells were increased, and the expression of NLRP3, caspase-1 and ASC was up-regulated in group EKO+ SAE ( P<0.05). Compared with group WT+ SAE, the time of staying at target quadrant was significantly shortened, the recognition index and cell survival rate were decreased, the contents of IL-1β, IL-18 and TNF-α and the number of NLRP3 positive cells were increased, and the expression of NLRP3, caspase-1 and ASC was up-regulated in group EKO+ SAE ( P<0.05). There was no significant difference in total distance moved between the four groups ( P>0.05). Conclusion:Erbin can exert endogenous protection by inhibiting the activation of NLRP3 inflammasomes in mice with SAE.
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Objective:To evaluate the effect of esketamine on acute kidney injury (AKI) in the rats with sepsis and the role of autophagy.Methods:Forty SPF healthy adult male Sprague-Dawley rats, weighing 200-240 g, were divided into 5 groups ( n=8 each) by a random number table method: control group (Con group), esketamine group (Con+ Ket group), sepsis group (lipopolysaccharide [LPS] group), sepsis plus esketamine group (LPS+ Ket group), and sepsis plus esketamine plus 3-methyladenine (3MA) group (LPS+ Ket+ 3MA group). The model of AKI was established by intraperitoneal injection of LPS in anesthetized rats.Normal saline 10 ml/kg was intraperitoneally injected in Con group.In Con+ Ket group, normal saline 10 ml/kg was intraperitoneally injected, and 30 min later esketamine 10 mg/kg was injected via the tail vein.LPS 10 mg/kg was intraperitoneally injected in LPS group.In LPS+ Ket group, LPS 10 mg/kg was intraperitoneally injected, and 30 min later esketamine 10 mg/kg was injected via the tail vein.In LPS+ Ket+ 3MA group, LPS 10 mg/kg was intraperitoneally injected, and 30 min later esketamine 10 mg/kg and 3-MA 15 mg/kg were injected via the tail vein.The rats were anesthetized at 24 h after intraperitoneal injection of LPS and then sacrificed, and renal tissues were removed for microscopic examination of the pathological changes which were scored and for determination of contents of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1, interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay) and expression of LC3, P62 and Beclin-1 (by Western blot). Results:Compared with Con group, the score for pathological damage to renal tissues and contents of NLRP3, ASC, caspase-1, IL-1β and IL-18 were significantly increased, LC3-Ⅱ/LC3-Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, and the expression of P62 was up-regulated in LPS group ( P<0.05). Compared with LPS group, the score for pathological damage to renal tissues and contents of NLRP3, ASC, caspase-1, IL-1β and IL-18 were significantly decreased, LC3-Ⅱ/LC3-Ⅰ ratio was increased, the expression of Beclin-1 was up-regulated, and the expression of P62 was down-regulated in LPS+ Ket group ( P<0.05). Compared with LPS+ Ket group, the score for pathological damage to renal tissues and contents of NLRP3, ASC, caspase-1, IL-1β and IL-18 were significantly increased, LC3-Ⅱ/LC3-Ⅰ ratio was decreased, the expression of Beclin-1 was down-regulated, and the expression of P62 was up-regulated in LPS+ Ket+ 3MA group ( P<0.05). Conclusion:Esketamine can reduce AKI and autophagy is involved in the process, which is related to inhibiting the activation of NLRP3 inflammasomes and decreasing inflammatory responses in rats with sepsis.
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Objective:To evaluate the role of ErbB2-interacting protein (Erbin) in muramyl dipeptide (MDP)-induced inflammatory responses in the macrophages of mice.Methods:Erbin gene knockout RAW264.7 cell line (Erbin -/ -RAW264.7) was constructed by CRISPR/CAS9 gene-editing technology.RAW264.7 cells were cultured in vitro.Each type of cells was divided into 2 groups ( n=16 each)by a random number table method: RAW264.7 group, RAW264.7 plus MDP group, erbin -/ -RAW264.7 group, and erbin -/ -RAW264.7 plus MDP group.In each MDP group, cells were incubated with 10 μg/ml MDP for 6 h, then immunofluorescence was used to determine the expression of nuclear factor kappa B (NF-κB) p65, and the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the culture medium were determined by enzyme-linked immunosorbent assay. Results:Compared with RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased( P<0.05), NF-κB p65 moved to the nucleus, and the red fluorescence area was increased in RAW264.7+ MDP group.Compared with RAW264.7+ MDP group and Erbin -/- RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased ( P<0.05), NF-κB p65 moved more markedly to the nucleus, and the red fluorescence area was increased in Erbin -/-RAW264.7+ MDP group. Conclusion:Erbin inhibits MDP-induced inflammatory responses in macrophages through inhibiting the activity of NF-κB p65 in mice.
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Objective To evaluate the effect of penehyclidine hydrochloride (PHC) on the expression of caveolin-1 (Cav-1) with lipopolysaccharide (LPS)-induced lung injury (LI) in rats.Methods Thirty SPF healthy male Sprague-Dawley rats,weighing 170-190 g,were divided into 3 groups (n =10each) using a random number table method:control group (group C),LPS-induced LI group (group LI)and PHCD group.LI was produced by injecting LPS 0.2 ml (5 mg/kg) via the trachea in anesthetized rats.PHCD 0.5 ml (2 mg/kg) was intraperitoneally injected at 1 h before establishing the model in group PHCD.Arterial blood samples were collected at 24 h after establishing the model for blood gas analysis and for determination of serum tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) concentrations by enzyme-linked immunosorbent assay.Rats were then sacrificed,and the lungs were removed.The main bronchus was lavaged,and the broncho-alveolar lavage fluid (BALF) was collected for calculation of the percentage of polymorphonuclear neutrophils (PMNs).Lung tissues were obtained for examination of pathological changes and for determination of myeloperoxidase (MPO) activity (by colorimetric assay),wet/dry weight ratio (W/D ratio),and expression of Cav-1 and nuclear factor kappa B (NF-sB) in nucleoprotein (by Western blot).Results Compared with group C,pH value and PaO2 were significantly decreased,the PaCO2,percentage of PMNs in BALF,W/D ratio and MPO activity were increased,the Car-1 expression was down-regulated,the expression of NF-κB in nucleoprotein was up-regulated,and the serum TNF-α and IL-1β concentrations were increased in group LI (P＜0.05).Compared with group LI,pH value and PaO2 were significantly increased,the PaCO2,percentage of PMNs in BALF,W/D ratio and MPO activity were decreased,the Cav-1 expression was up-regulated,the expression of NF-κB in nucleoprotein was down-regulated,and the serum TNF-α and IL-1β concentrations were decreased (P＜0.05),and the path ological changes of lung tissues were significantly attenuated in group PHCD (P＞0.05).Conclusion The mechanism by which PHC reduces LPS-induced LI may be related to up-regulating the expression of Cav-1 and mitigating inflammatory responses in lung tissues of rats.
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Objective To evaluate the relationship between cholinergic anti-inflammatory pathway and autophagy during liver injury in septic mice.Methods SPF healthy male 32 C57BL/6 mice,aged 6-8 weeks,weighing 18-22 g,were divided into 4 groups (n=8 each) using a random number table method:sham operation group (group Sham),sepsis group (group Sep),α7nAChR agonist GTS-21 plus sepsis group (GTS-21+Sep group),and α7nACh antagonist α-BGT plus sepsis plus GTS-21 group (α-BGT+Sep+GTS-21 group).Sepsis was induced by cecal ligation and puncture.GTS-21 4 mg/kg was intraperitoneally injected immediately after operation in group GTS-21 +Sep.α-BGT 1 mg/kg was intraperitoneally injected at 15 min before operation,and GTS-21 4 mg/kg was intraperitoneally injected immediately after operation in group α-BGT+Sep+GTS-21.Blood samples were obtained at 6 h after surgery for determination of serum aspartate transaminase (AST) and alanine transaminase (ALT) concentrations by enzyme-linked immunosorbent assay.Liver tissues were obtained for determination of the expression of microtubule-associated protein-1 light chain 3-Ⅱ (LC3 Ⅱ) and sequestosome-1/p62 (by Western blot) and for examination of the number of autophagosomes in liver cells (under a transmission electron microscope).Results Compared with Sham group,the expression of LC3 Ⅱ and sequestosome-1/p62 was significantly up-regulated,the number of autophagosomes was increased,and the serum AST and ALT concentrations were increased in Sep group (P＜0.05).Compared with Sep group,the expression of LC3 Ⅱ was significantly up-regulated,the expression of sequestosome-1/p62 was down-regulated,the number of autophagosomes was increased,and the serum AST and ALT concentrations were decreased in GTS-21 +Sep group (P＜0.05).Compared with GTS-21 +Sep group,the expression of LC3 Ⅱ was significantly down-regulated,the expression of sequestosome-1/p62 was up-regulated,the number of autophagosomes was decreased,and the serum AST and ALT concentrations were increased in α-BGT+Sep+GTS-21 group (P＜0.05).Conclusion Activation of the cholinergic anti-inflammatory pathway can enhance the level of autophagy in liver cells and is involved in the endogenous protective mechanism of sepsis-induced liver injury in septic mice.
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Objective To evaluate the relationship between inflammatory responses and autophagy in lung tissues after scald and nucleotide-binding oligomerization domain-containing protein 2 (NOD2) signaling pathway in septic rats.Methods Twenty SPF healthy male Sprague-Dawley rats,weighing 200-250 g,were divided into control group (group C,n=10) and sepsis after scald group (group SS,n=10) using a random number table.The rats were subjected to a third-degree scald burn covering 20％ of total body surface area (body surface was shaved and then exposed to 99-100 ℃ water for 12 s),and 24 h later muramyldipeptide 5 mg/kg was intravenously injected to induce sepsis.The rats were only exposed to 20 ℃ water,and 24 h later normal saline 1 ml was given instead in group C.At 6 h after muramyldipeptide injection in group SS and at 6 h after normal saline injection in group C,arterial blood samples were collected for determination of serum tumor necrosis factor-r and interleukin-6 concentrations by enzyme-linked immunosorbent assay.Then rats were sacrificed and lungs were removed tor measurement of activity of myeloperoxidase,NOD2 mRNA expression (using real-time polymerase chain reaction) and expression of receptor interacting protein 2,nuclear factor kappa Bp65 and microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ) and LC3 Ⅱ in lung tissues (by Western blot).The LC3 Ⅱ / Ⅰ ratio was calculated.Results Compared with group C,the expression of NOD2 mRNA,receptor interacting protein 2 and nuclear factor kappa Bp65 was significantly up-regulated,and the LC3 Ⅱ / Ⅰ ratio and serum tumor necrosis factor-α and interleukin-6 concentrations were increased in group SS (P＜0.05).Conclusion The mechanism underlying enhanced inflammatory responses and autophagy in lung tissues during sepsis after scald may be related to activation of NOD2 signaling pathway in rats.
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Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.
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Objective To evaluate the effect of penehyclidine hydrochloride on cell apoptosis during acute lung injury (ALI) induced by two-hit in rats.Methods Thirty male SPF Sprague-Dawley rats, aged 8 weeks, weighing 240-270 g, were randomly assigned into 3 groups (n =10 each) using a random number table: sham operation group (group Sham), ALI induced by blunt chest trauma and hemorrhagic shock group (group ALI), and penehyclidine hydrochloric group (group PHC).In ALI and PHC groups, the rats were subjected to the combination of chest trauma and hemorrhage (mean arterial pressure 35-40 mmHg mm Hg, lasting for 60 min) to establish a model of ALI.In group PHC, penehyclidine hydrochloric 2 mg/kg was injected intraperitoneally at 1 h before blunt chest trauma.At 8 h after successful establishment of the model, the rats were sacrificed, and lungs were removed for examination of the pathologic changes and for determination of Bax, Bcl-2 and caspase-3 expression (using Western blot), tumor necrosis factor-alpha (TNF-α) content (by enzyme-linked immunosorbent assay) , and cell apoptosis (by TUNEL).Apoptotic index was calculated.Results Compared with group Sham, the levels of Bax, caspase-3 and TNF-α and apoptotic index were significantly increased, and Bcl-2 expression was down-regulated in ALI and PHC groups.Compared with group ALI, the levels of Bax, caspase-3 and TNF-α and apoptotic index were significantly decreased, and Bcl-2 expression was up-regulated in group PHC.The pathologic changes of lungs were significantly reduced in group PHC than in group ALI.Conclusion Penehyclidine hydrochloride mitigates ALI induced by two-hit through inhibiting cell apoptosis in rats.
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Objective To evaluate the effect of electro-acupunctare at Zusanli on liver injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C); sepsis after scald group (group SS); electro-acupuncture at Zusanli group (group E); electric stimulation of non-acupoint group (group NE); electro-acupuncture at Zusanli + α-bungarotoxin (α-BGT) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ of the total body surface area (TBSA) and sepsis was induced with muramyl dipeptide (MDP) 5 mg/kg injected into the femoral vein immediately after scald.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral Zusanli acupoints or non-acupoints was performed for 12 min every 8 h for 2 consecutive days starting from the time point immediately after MDP was injected in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the acupoints of bilateral Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in normal saline 1 ml) was injected into the femoral vein before electric stimulation,then electro-stimulation was performed and the method was similar to those previously described in group E.After 48 h of continuous stimulation,liver specimens were obtained for microscopic examination of the pathological changes.The blood samples were obtained from the abdominal aorta for determination of the levels of serum ALT and AST,tumor necrosis factor-alpha (TNF-α),and high mobility group box-1 (HMGB1) (by ELISA),and expression of Nod-like receptor 2 (NLR2) mRNA (by RTPCR) and receptor-interacting protein 2 (RIP2) in the liver tissues (by Western blot).Results The serum ALT and AST,TNF-α and HMGB-1 levels and NLR2 mRNA and RIP2 expression in liver tissues were significantly higher in SS group than in C group.Compared with SS group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly decreased and NLR2 mRNA and RIP2 expression was down-regulated in E group,and no significant changes were found in the parameters mentioned above in NE group.Compared with E group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly increased,and NLR2 mRNA and RIP2 expression was up-regulated in α-BGT group.The pathological changes in liver tissues were significantly reduced in group EA as compared with SS,NE and α-BGT groups.Conclusion Electro-acupuncture at Zusanli can reduce liver injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic antiinflammatory pathway mediated by α7 nicotinic acetylcholine receptors in liver tissues may be involved in the mechanism.
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Objective To investigate the effect of electro-acupunctare at zusanli on acute lung injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C),sepsis after scald group (group SS),electro-acupuncture at zusanli group (group E),electric stimulation of non-acupoint group (group NE) and electro-acupuncture at zusanli + α-bungarotoxin (α-BGT,a selective α7 nicotinic acetylcholine receptor antagonist) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ total body surface (TBS) and muramyl dipeptide (MDP) 5 mg/kg was injected into the femoral vein immediately after scald to induce sepsis.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral zusanli was performed for 12 min starting from the time point immediately after MDP injection and every 8 h for 2 consecutive days in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the bilateral acupoints of Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in 1 ml of normal saline) was injected into the femoral vein before electro-stimulation of zusanli.At 48 h after treatment,arterial blood samples were obtained for determination of serum tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) protein levels (by ELISA) and lung specimens were removed for microscopic examination and for determination of the expression of Nod like receptor 2 (NLR2) mRNA (by RT-PCR) and receptor interacting protein 2 (RIP2) in the lung tissues (by Western blot).Results Compared with group C,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in SS,NE and α-BGT groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group E (P ＞ 0.05).Compared with group SS,the expression of NLR2 mRNA and RIP2 was down-regulated,and the serum TNF-α and HMGB1 levels were decreased in group E (P ＜ 0.05),and no significant change was found in the parameters mentioned above in NE and α-BGT groups (P ＞ 0.05).Compared with group E,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in NE and α-BGT groups (P ＜ 0.05).The pathological changes of lung tissues were significantly reduced in group E as compared with group SS.Conclusion Electro-acupunctare at Zusanli can reduce acute lung injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic anti-inflammatory pathway in lung tissues may be involved in the mechanism.
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Objective To evaluate the regulatory role of acetylcholine receptor in muramyl dipeptide (MDP)-induced activation of Nod-like receptor 2/receptor-interacting protein 2 (2NLR2/RIP2) pathway in macrophages of mice.Methods RAW264.7 cells at the logarithmic growth phase were seeded in 12-well plates (density 1 × 106 cells/ml,2 ml/well),a total of 108 wells.The cells were randomly divided into 3 groups (n =36 each) using a random number table:control group (group C),MDP group (group M),and GTS-21 (a7nAChR specific agonist) group (group G).The cells were routinely cultured in group C.MDP with the final concentration of 10 μg/ml was added to the culture medium in group M.MDP with the final concentration of 10μg/ml and GTS21 with the final concentration of 50 μg/ml were added to the culture medium in group G.The cells were incubated for 24 h.At 1,6 and 24 h of incubation with MDP,12 wells were chosen and the cell suspension was obtained for measurement of NLR2 mRNA expression (by real-time fluorescent quantitative PCR),RIP2 expression (by Western blot),and concentrations of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) in the culture media (by ELISA).Results Compared with group C,the levels of NLR2 mRNA,RIP2,TNFα and HMGB1 were significantly increased at each time point in group M (P ＜ 0.05).Compared with group M,the levels of NLR2 mRNA,RIP2,TNF-α and HMGB1 were significantly decreased at each time point in group G (P ＜ 0.05).Conclusion Acetylcholine receptor can suppress MDP-induced transduction of NLR2/RIP2 pathway in macrophages of mice.
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Objective To establish a rat model of sepsis induced by muramyl dipeptide (MDP) after scald burn.Methods Fifty SPF male Sprague-Dawley rats,aged 2-3 months,weighing 200-250 g,were randomly divided into 3 groups:control group (group C,n =10),scald group (group S,n =10) and MDP group (n =30).The rats were subjected to a third-degree scald burn covering 20% of total body surface area in groups S and MDP.The rats were only exposed to 20 ℃ water in group C.MDP 5 mg/kg was injected via the femoral vein at 24 h after scald bum in group MDP.Arterial blood samples were collected at 1,6 and 24 h after MDP injection in group MDP,at 24 h after scald burn in group S,or at 24 h after exposure to 20 ℃ water in group C for blood gas analysis and for measurement of white blood cell (WBC) and platelet (Plt) counts,serum aminotransferase (ALT),aspartate transferase (AST),total bilirubin (TB),creatinine (Cr) and blood urea nitrogen (BUN) levels,creatine kinase isoenzyme-MB (CK-MB) activity,and plasma tumor necrosis factor-α (TNF-α),interferon-γ(IFN-γ),interleukin-6 (IL-6),IL-10 and high mobility group box 1 protein (HMGB-1) levels.The rats were sacrificed after collecting blood samples,and heart,liver,lung,and kidney specimens were obtained for microscopic examination of pathologic changes.The activity of myeloperoxidase (MPO) in lung tissues was measured.Another 90 male Sprague-Dawley rats were randomly divided into 3 groups and treated as the method previously described for record of the survival rate within 72 h.Results Compared with C group,the plasma IL-6,IL-10,IFN-γ and HMGB1 levels,WBC count,serum ALT,AST,and BUN levels and MPO activity were significantly increased,and the survival rate within 72 h was decreased in S group,and the plasma TNF-α,IL-6,IL-10,IFN-γand HMGB-1 levels,serum ALT,AST,TB,BUN,Cr and CK-MB levels,MPO activity,PaCO2 and BE value were significantly increased,and WBC and PLT counts,pH value,PaO2 and survival rate within 72 h were decreased in MDP group (P ＜ 0.05).Compared with S group,the plasma TNF-α,IL-6,IFN-γ and HMGB-1 levels,serum ALT,AST,TB,BUN,Cr and CK-MB levels,MPO activity,PaCO2 and BE value were significantly increased,and WBC and Plt counts,pH value,PaO2 and survival rate within 72 h were decreased in MDP group (P ＜ 0.05).The pathologic changes of heart,liver,lung and kidney were obvious in S and MDP groups and severer in MDP group.Conclusion After a third-degree 20% total body surface area scald burn,MDP induces excessive production of inflammatory cytokines accompanying with multiple organ damage ; thus the model of sepsis is successfully established after scald burn in rats.
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Objective To. investigate the effect of electroacupuncture at acupoint Zusanli (ST36) on the liver injury during the early stage after bum in rats. Methods Forty adult male SD rats weighing 220-250 g were randomly divided into 5 groups ( n = 8 each) : group sham operation (group Ⅰ ) ; group burn (group Ⅱ ) ; group acupoint at Zusanli (ST36) (group Ⅲ ); group non-acupoint stimulation (group Ⅳ ) and group ST36 + alphabungarotoxin (alpha7 nicotinic acetylcholine receptor antagonist) (group Ⅴ ). Rats were subjected to 3rd degree burn covering 30% of the total body surface area. Rats were resuscitated with lactataed Ringer's solution according to Parkland formula (4 ml/kg per 1% body surface area) immediately after burn. Bilateral acupoints Zusanli were stimulated with constant voltage (3 V, 3 Hz,2ms) for 20 min 3 times a day for 2 days starting immediately after resuscitation in H and V groups. In group V alpha-bungarotoxin 1.0 μg/kg was administered iv immediately after fluid resuscitation before acupuncture. In group Ⅳ same electric stimulation was performed at a point 0.5 cm lateral to Zusanli. The animals were sacrificed at 48 h after burn. The content and expression of high mobility group box 1 (HMGB1) protein in liver were measured. Liver specimens were obtained for microscopic examination (with light and electronic microscope). Results Compared with group Ⅰ , hepatic HMGB1 protein level significantly increased in Ⅱ and Ⅳ groups. There were significant ultrastructural changes in the liver in burn rats in group Ⅱ and group Ⅳ. Electric stimulation of ST36 significantly attenuated the histologic changes in the liver and decreased the hepatic HMGB1 protein level in group Ⅲ . Pretreatment with specific alpha.7 nicotinie acetylcholine receptor antagonist alpha-bungarotoxin reversed the beneficial effect of electroacupuncture at Zusanli. Conclusion Electric stimulation of acupoint ST36 can ameliorate liver injury during the early stage of burn by activating alpha7 nicotinic acetylcholine receptor-mediated pathways for anti-inflammation.
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Objective To investigate the effect of nicotine on coagulation abnormalities in endotoxemic rats.Methods Ninety-six male SD rats weighing 200-250 g were randomly divided into 4 groups (n=24 each): group normal saline (group NS);group LPS;group nicotine(group NIC)and group α-bungarotoxin (α7 nicotinic acetylcholine receptor antagonist, group α-BGT) . Endotoxemia was induced by LPS 10 mg/kg injected via femoral vein in LPS, NIC and α-BGT groups. In group NIC nicotine 400 μg/kg was injected intraperitoneally at 30 min before LPS injection. In group α-BGT α-BGT 1 μg/kg was injected intraperitoneally at 15 min before intraperitoneal nicotine. Prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(Fib),antithrombin (AT),von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1),D-dimer,platelet count and TNF-α were measured before (baseline) and 2, 4 and 6 h after LPS injection.Results PT and APTT were significantly prolonged and plasma Fib and AT concentrations and platelet count were significantly decreased, while plasma PAI-1, D-dimer, vWF and TNF-α concentrations were significantly increased after LPS administration in group LPS as compared with group NS. Nitotine pretreatment significantly attenuated the LPS-induced changes in group NIC.The effect of nicotine was counteracted by α-BGT. Conclusion Nicotine can attenuate coagulation abnormalities induced by LPS by acting on α7 nicotinic acetylcholine receptor.
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Objective To investigate the effect of electro-acupuncture at zusanli on Severe thermal injury-induced acute lung injury in rats.Methods Forty male SD rats weighing 200-250 g were used in this study.Thirty percent of the total body surface (TBS) was shaved chemically with 20% sodium sulfate and then exposed to 99-100℃ water for 12 s.The animals with third degree thermal injury involving 30% TBS were randomly divided into 5 groups(n=8 each):group Ⅰ control(group C);groupⅡ thermal injury;group Ⅲ electro-acupuncture at zusanli;group Ⅳ electric stimulation of non-acupoint and group Ⅴ electro-acupuncture at zusanli+α-bungarotoxin α-BGT).In group Ⅲ,Ⅳ,and Ⅴ electro-stimulation(3 v,2 ms,3 Hz) of zusanli or non-acupoint was performed for 12 min immediately after thermal injury model was established and every 8 h.hung specimens were obtained at 48 h after thermal injury for microscopic examination.The pulmonary HMGB-l protein level was measured by ELISA.The expression of HMGB-1 mRNA and protein in the lung was determined by RT-PCR and immuno-histochemistry respectively.Results Thermal injury induced leucocytosis in the interstitial capillaries,interstitial edema,intra-alveolar fibrin deposit,blebbing of type Ⅱ alveolar lining cells and decrease in lamellar body.Both expression of HMGB-1 mRNA and protein in the lung was significantly enhanced at 48 h after thermal injury.Electrical stimulation of zusanli significantly down-regalated the expression of HMGB-1 mRNA and protein in the lung.However,α-BGT pretreatment reversed the effects of electrical stimulation of zusanli.Conclusion Electrical stimulation of zusanli could significantly ameliorate severe thermal injury-induced acute lung injury through inhibition of HMGB-1 mRNA and protein expression and activation of cholinergic anti-inflammatory pathway mediated by nicotinic acetylcholine receptor α7 subunit.
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Objective To identify the correlation factors associated with cerebral intragenic ischemia after temporary parent arterial occlusion in intracranial aneurysm surgery. Methods One hundred and eighteen patients who underwent temporary arterial occlusion in the 120 aneurysms (from a group of 324 consecutive aneurysm patients treated from 1996 to 2002) were reviewed retrospectively. These variables included sex, age, presence of preoperative subarachnoid hemorrhage (SAH), neurological clinical grade, operational timing, duration of arterial occlusion, numbers of temporary occlusion, mode of arterial occlusion, intraoperative aneurysm rupture, hypertension, the location of temporary occlusion, aneurysm size, hyperglycemia, atheromatous mass. Univariate and multivariate were used to investigate the relationship between the variates and postoperative ischemic changes. Results The total times of temporary occlusion were 156, with an average of 1.30. The duration of arterial occlusion ranged from 1 to 45 min (9.75?7.75). Seventeen patients (14.4%) demonstrated evidence of new infarction in the vascular territory subjected to temporary arterial occlusion. Conclusion In the univariate analysis, age, presence of preoperative SAH, duration of arterial occlusion, atheromatous mass are all significantly correlated with postoperative ischemic injuries. Multivariate logistic regression revealed that the age, older more than 60 (P= 0.010 3 , relative risk=4.335), and the duration of arterial occlusion, lasting more than 20 min (P= 0.032 9 , relative risk=4.177), have significant correlation with the injuries. Based on these findings, temporary occlusion is safe and useful in aneurysm surgery and the postoperative cerebral ischemia is less likely to occur when the duration of clipping is shorter than 20 min.
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Objective To investigate the effects of ketamine on endotexin-induced production of proinflammatory cytokines(IL-6, TNF-?) and activation of their medulating factor NF-?B in vivo. Methods Forty adult male SD rats weighing 200-250 g were randomly divided into 4 groups: (Ⅰ)control group(n=10); (Ⅱ) endotoxin group received intravenous endotoxin(Escherichia coli O111: B4, Sigma) 5 mg?kg~(-1)(n=10); (Ⅲ, Ⅳ)endotexin+ketamine group received ketamine or 5 or 10mg?kg~(-1)?h~(-1) after endotoxin(n=10). The animals were anesthetized with urethane i.p. (1g?kg~(-1)). Carotid artery was cannulated for BP and HR monitoring and jugular vein was cannulated for fluid or drug administration. Two hours after endotoxin administration the animals were sacrificed by exsanguination. Blood was collected and peripheral blood monocytes(PBMC) were isolated. NF-?B activity in PBMC was measured by EMSA and plasma TNF-? and IL-6 levels were determined by ELISA. Results Progressive hypotension and tachycardia developed after endotoxin administration. Endotexin also increased NF-?B activity in PBMCs and plasma TNF-? and IL-6. Ketamine 10 mg?kg~(-1) attenuated the endotexin-induced hemedynamie levels. Ketamine(5, 50 mg?kg~(-1)?h~(-1)) suppressed NF-?B activity in PBMC and inhihited plasma TNF-? level but plasma IL-6 level was not affected. Conclusion Ketamine can suppress endotoxin-induced NF-?appa B activation. Subanesthetic dose of ketamine has anti-inflammatory action.