1.Effects of complement C1q on motor function and postsynaptic density protein 95 of rats with cere-bral ischemia-reperfusion injury
Qingtong ZHANG ; Chenyao JIAN ; Mingda LIU ; Shenquan CAI ; Manlin DUAN
The Journal of Clinical Anesthesiology 2024;40(8):855-859
Objective To explore the effects of complement C1q on postsynaptic density protein 95(PSD95)and motor function of rats with cerebral ischemia-reperfusion injury.Methods Thirty-six clean male SD rats,aged 6-8 weeks,weighing 220-250 g,were randomly divided into three groups:sham group(group S),middle cerebral artery occlusion(MCAO)group(group M),and MCAO+C1q neutralization group(group A),12 rats in each group.Group S only received anatomical separation of common carotid ar-tery without insertion of monofilament,group M established the MCAO model was prepared by insertion of monofilament,and group A received C1q neutralizing antibody 10 μl injected into the lateral ventricle on the first day after establishing MCAO model.Three days after modeling,the adhesion removal experiment was used to record the adhesion stimulus removal time,and the balance beam experiment was used to evaluate the limb motor function of the rats.After the rats were sacrificed,the ELISA method was used to detect the C1q and C3 levels in the hippocampus tissue,the Western blot method was used to detect the C1q subcom-ponent subunit A(C1qa)and PSD95 protein levels in the hippocampus tissue,and the Nissl staining was used to record the number of Nissl bodies.Results Compared with group S,group M exhibited a signifi-cantly longer adhesion removal time,significantly elevated balance beam experiment score,significantly higher levels of C1q,C3,and C1qa in the hippocampus,and significantly lower levels of PSD95 and Nissl's body count in the hippocampus(P<0.05).Compared with group M,group A exhibited significantly shor-ter adhesion removals,a significantly lower balance beam experiment score,significantly lower levels of C1q,C3,and C1qa in the hippocampus,significantly higher PSD95 content,and a significantly higher Nissl's body count in the hippocampus(P<0.05).Conclusion Cerebral ischemia-reperfusion injury in rats can induce extensive activation of the complement system and aggravate the loss of synaptic protein PSD95 and motor dysfunction in rat hippocampal tissue through complement protein C1q.Neutralizing com-plement therapy can effectively improve cerebral ischemia-reperfusion injury in rats.
3.Effect of hydrogen on expression of hippocampal cold-inducible RNA-binding protein after cardiac arrest-resuscitation in rats
Lin WU ; Yingchun DONG ; Kangli HUI ; Miaomiao XU ; Baojian ZHAO ; Manlin DUAN
Chinese Journal of Anesthesiology 2020;40(4):477-480
Objective:To evaluate the effect of hydrogen on the expression of hippocampal cold-inducible RNA-binding protein (CIRP) after cardiac arrest-resuscitation in rats.Methods:Ninety clean-grade healthy male Sprague-Dawley rats, weighing 280-320 g, were randomly divided into 3 groups: sham group (group Sham, n=20), cardiac arrest-cardiopulmonary resuscitation group (group CPR, n=35), and hydrogen-rich saline group (group H 2, n=35). Cardiac arrest was induced by transoesophageal cardiac pacing followed by CPR in group CPR.Only femoral arteriovenous puncture and tracheal intubation were performed in group Sham.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected immediately after recovery of spontaneous circulation (ROSC) and at 6 and 12 h after ROSC in group H 2 , while the equal volume of normal saline was given instead in the other two groups.Neuro-functional deficit was assessed using neurologic deficit scores (NDS) at 1 and 3 days after ROSC.The animals were sacrificed immediately after intubation in group Sham and at 6 h and 1, 2 and 3 days after ROSC in CPR and H 2 groups, and the hippocampal tissues were obtained to detect the expression of nuclear and cytoplasmic CIRP by Western blot. Results:Compared with group Sham, NDS was significantly decreased at each time point after ROSC in group CPR and group H 2, the expression of nuclear CIRP was significantly down-regulated at 1, 2 and 3 days after ROSC, and the expression of cytoplasmic CIRP was up-regulated at 1 and 2 days after ROSC in group CPR, and the expression of nuclear CIRP was significantly down-regulated at each time point after ROSC, and the expression of cytoplasmic CIRP was down-regulated at 2 and 3 days after ROSC in group H 2 ( P<0.05). Compared with group CPR, NDS was significantly increased at each time point after ROSC, the expression of nuclear CIRP was down-regulated at 6 h after ROSC, and the expression of cytoplasmic CIRP was down-regulated at 1 and 2 days after ROSC in group H 2 ( P<0.05). Conclusion:The nechanism by which hydrogen reduces brain injury after cardiac arrest-resuscitation may be related to down-regulating hippocampal CIRP expression in rats.
5.Effect of mild hypothermia combined with hydrogen-rich saline on cerebral injury after cardiac arrest and resuscitation in rats
Yu GAO ; Zhengqin SUN ; Qiang WANG ; Manlin DUAN ; Jianguo XU
Chinese Journal of Anesthesiology 2019;39(5):556-560
Objective To evaluate the effect of mild hypothermia combined with hydrogen-rich saline on cerebral injury after cardiac arrest and resuscitation in rats.Methods Healthy male Sprague-Dawley rats,aged 7-8 weeks,weighing 280-320 g,were divided into 5 groups (n=33 each) using a random number table method:sham operation group (group S),cardiac arrest and resuscitation group (group CAR),hydrogen-rich saline group (group H2),mild hypothermia group (group MH),and mild hypothermia plus hydrogen-rich saline group (group MH+H2).Cardiac arrest was induced with transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the cerebral injury model.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected immediately after return of spontaneous circulation (ROSC) in H2 and MH+H2 groups,while the equal volume of normal saline was given instead in the other groups.The body temperature of rats was cooled down to 32-34℃ within 15 min starting from the time point immediately after ROSC and maintained for 4 h in MH and MH+H2 groups.Fifteen rats were selected at 24 h after ROSC to assess the neurological function score (NDS).Eighteen rats in each group were sacrificed at 24 h after ROSC,and brains were removed for microscopic examination of the pathological changes in hippocampal CA1 region after hematoxylin and eosin staining and for determination of pyramidal cell count and expression of glucose-regulated protein 78 (GRP78),C/EBP-homologous protein (CHOP),caspase-12,caspase-3,Bcl-2 and Bax in hippocampal CA1 region (by Western blot).Results Compared with group S,the NDS was significantly decreased,the pyramidal cell count was reduced,the expression of GRP78,CHOP,caspase-12,caspase-3 and Bax was up-regulated,and the expression of Bcl-2 was down-regulated in the other four groups (P<0.05).Compared with group CA-R,the NDS and pyramidal cell count were significantly increased,the expression of GRP78 and Bcl-2 was up-regulated,and the expression of CHOP,caspase-12,caspase-3 and Bax was down-regulated in H2,MH and MH+H2 groups (P<0.05).Compared with group H2 or group MH,the NDS and pyramidal cell count were significantly increased,the expression of caspase-3 and Bax was down-regulated,the expression of Bcl-2 was up-regulated (P<0.05),and no significant change was found in the expression of GRP78,CHOP and caspase-12 in group MH+H2 (P> 0.05).Conclusion Combination of mild hypothermia and hydrogen-rich saline offers enhanced efficacy in reducing cerebral injury after cardiac arrest and resuscitation over mild hypothermia or hydrogen-rich saline alone in rats.
6.Changes in expression of cold-inducible RNA-binding protein in hippocampus during brain injury in a rat model of cardiac arrest-cardiopulmonary resuscitation
Yongyi CHEN ; Manlin DUAN ; Minhao ZHANG ; Liangbin CAO ; Juan LI ; Jiejie ZHOU ; Lianbing GU
Chinese Journal of Anesthesiology 2018;38(5):602-605
Objective To evaluate the changes in expression of cold-inducible RNA-binding protein (CIRP) in hippocampus during brain injury in a rat model of cardiac arrest-cardiopulmonary resuscitation.Methods Seventv-two clean-grade healthy male Sprague-Dawley rats,weighing 280-350 g,aged 8-10 weeks,were divided into 2 groups using a random number table:sham operation group (S group,n=18) and ischemia-reperfusion group (I/R group,n=54).Tracheal intubation was performed and artery and veins were punctured in group S.Ventricular fibrillation was induced by transoesophageal cardiac pacing to establish the model of cardiac arrest in group I/R.Rats were sacrificed at 12,24 and 48 h after resuscitation and the hippocampus was harvested for determination of CIRP,tumor necrosis factor-alpha (TNF-α) and interleukin-lbeta (IL-1β) protein and mRNA expression (by quantitative polymerase chain reaction or Western blot) and for determination of pathological changes of hippocampi (with a light microscope).Results Compared with group S,the expression of CIRP mRNA in hippocampus was up-regulated at 24 and 48 h after resuscitation,the expression of TNF-α mRNA was up-regulated at 12,24 and 48 h after resuscitation,the expression of IL-1β mRNA was up-regulated at 12 and 24 h after resuscitation,and the expression of CIRP,TNF-α and IL-1β was up-regulated at 12,24 and 48 h after resuscitation in group I/R (P<0.05).Pathological changes in hippocampal CA1 region were found in group I/R.Conclusion The expression of CIRP in hippocampus is up-regulated,which promotes central inflammatory responses during brain injury in a rat model of cardiac arrest-cardiopulmonary resuscitation.
7.A study on the safety of pneumoperitoneum performed under intravenous anesthesia during robotic rectectomy
Li TANG ; Qian LI ; Manlin DUAN ; Dunyi QI
Journal of Chinese Physician 2018;20(11):1651-1655
Objective The purpose of this study was to assess the safety of carbon dioxide pneumoperitoneum on patients during robotic rectectomy.Methods 50 patients [American Society of Anesthesiologists (ASA) physical status Ⅱ,18 and 65 years of age] underwent rectal cancer surgery were selected in our study.According to whether or not robotic assisted surgery was performed,they were divided into a robotic surgery group (group RS) and a laparoscopic surgery group (group LS) by surgical approach,25 patients in each group.All participants were given the same anesthesia protocol.Arterial blood samples 1 ml was obtained from the left radial artery for blood gas analysis to measure the partial pressure of arterial carbon dioxide (PaCO2) and and calculate the arterial-to-end-tidal carbon dioxide pressure difference (Pa-ETCO2) just 10 min after endotracheal intubation (T0),at 30 min(T1),1 h(T2),2 h(T3) after pneumoperitoneum and 30 minutes after release (T4).Meanwhile,the airway peak pressure was monitored.Blood samples (4 ml) extracted at T0,T3 and T4 were centrifuged and measured the serum levels of interleukin (IL)-6 and IL-10 by enzyme linked immunosorbent assay (ELISA).The time to resuscitation,extubation time,intraoperative medication and perioperative adverse events were all recorded.Results Compared with group LS,PaCO2 in the RS group was increased significantly at T1,T2 and T3 after pneumoperitonum and the IL-6 was lower at T4 (P < 0.05).There were no statistically significant differences in Pa-ETCO2,airway peak pressure,IL-10,time to resuscitation,extubation time and the incidence of adverse events between the two groups (P > 0.05).Conclusions It is safe for normal adult patients performed by intravenous anesthesia during robotic-assisted rectal surgery and the inflammatory response is small,which is beneficial to the patient's postoperative recovery.
8.Effect of hydrogen sulfide on hippocampal endoplasmic reticulum stress during global cerebral ischemia-reperfusion in rats
Liangbin CAO ; Yongyi CHEN ; Shenquan CAI ; Jingjing FAN ; Hao ZHONG ; Manlin DUAN ; Jianguo XU
Chinese Journal of Anesthesiology 2017;37(9):1091-1094
Objective To evaluate the effect of hydrogen sulfide on hippocampal endoplasmic reticulum stress during global cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two pathogen-free healthy male Sprague-Dawley rats,weighing 280-320 g,aged 8-10 weeks,were divided into 3 groups (n=24 each) using a random number table:sham operation group (group Sham),global cerebral I/R group (group I/R) and global cerebral I/R plus sodium hydrosulfide group (group I/R+NaHS).Cardiac arrest was induced with transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the global cerebral I/R model.Immediately after recovery of spontaneous circulation,sodium hydrosulfide 2.5 mg/kg was intraperitoneally injected in group I/R+NaHS,and normal saline 5 ml/kg was given in group I/R.The hippocampi were immediately removed at 24 h of reperfusion for determination of the expression of glucose-regulated protein 78 (GRP78),C/EBP-homologous protein (CHOP) and caspase-12 in hippocampal tissues (by Western blot).At 1,3 and 7 days of reperfusion,the hippocampal tissues were obtained and stained with haematoxylin and eosin for examination of the pathological changes in hippocampal CA1 region (under a light microscope) and for determination of apoptosis in hippocampal cells (using TUNEL staining),and the apoptosis rate was calculated.Results Compared with group Sham,the apoptosis rate of hippocampal tissues at 1,3 and 7 days of reperfusion in group I/R and at 3 and 7 days of reperfusion in group I/R+NaHS were significantly increased,and the expression of GRP78,CHOP and caspase-12 in hippocampal tissues was significantly up-regulated in I/R and I/R+NaHS groups (P<0.05).Compared with group I/R,the apoptosis rate of hippocampal tissues was significantly decreased,and the expression of GRP78,CHOP and caspase-12 was down-regulated at 1,3 and 7 days of reperfusion (P<0.05),and the pathological changes were significantly attenuated in group I/R+NaHS.Conclusion The mechanism by which hydrogen sulfide reduces apoptosis in hippocampal cells is related to inhibition of endoplasmic reticulum stress during global cerebral I/R in rats.
9.Effect of dexmedetomidine on proliferation, migration and invasion of human gastric cancer SGC-7901 cells
Jianhua TONG ; Ningning JI ; Manlin DUAN ; Zhiqiang ZHOU ; Ming XIA ; Jianguo XU
The Journal of Clinical Anesthesiology 2017;33(11):1096-1098
Objective To observe different behavior of proliferation,migration and invasion of SGC-7901 cells when exposured to dexrnedetomidine of different concentrations.Methods Human gastric cancer cells SGC-7901 were inoculated on culture plate for 24 h,then were randomly divided into 5 groups:control group (group C),dexmedetomidine 312.5μg/ml group (group D1),dexme detomidine 625μg/ml group (group D2),dexmedetomidine 1 250 μg/ml group (group D3),dexmedetomidine 2 500 μg/ml group (group D4).Each group was medicated and incubated for 48 h,then the cell proliferation,migration and invasion immediately were detected by CCK-8 and Transwell.Results SGC-7901 cell viability of groups D1,D2,D3 和 D4 had no significant difference compared with that of group C.The invasion ability and migration ability of SGC-7901 cells in groups D1,D2,D3 and D4 were significantly higher than those in group C (P < 0.05 or P < 0.01).Conclusion Dexmedetomidine can promote migration and invasion of SGC-7901 cells.
10.Effect of continuous incision infusion different concentration of ropivacaine for postoperative analgesia after radical mastectomy
Ming XIA ; Hui LI ; Jianhua TONG ; Ningning JI ; Wen QI ; Manlin DUAN ; Shaohua WANG ; Jianguo XU
The Journal of Clinical Anesthesiology 2017;33(1):19-21
Objective To observe the effect of continuous incision infusion different concentra-tion of ropivacaine for postoperative analgesia after radical mastectomy.Methods One hundred pa-tients under radical mastectomy,aged 40-70 years,ASA Ⅰ or Ⅱ,were randomly divided into four groups (n =25 each):0.2% (group R1),0.3% (group R2),0.4% (group R3)ropivacaine incision continued infiltration group and patient-controlled intravenous analgesia (group PCIA)as control group.VAS pain scores,sedation Ramsay score and side effects were recorded at each time point in rest and turning over 90°,2 h (T1 ),4 h (T2 ),8 h (T3 ),12 h (T4 ),24 h (T5 ),48 h (T6 )after the operation.Results VAS scores in group R1 at T1-T6 in rest and turn over 90°were significantly high-er than that of group PCIA (P <0.05).There were no significant differences among the group PCIA, group R2 and group R3.Sedation score in PCIA group was significantly higher than that in the other three groups (P <0.05),and the adverse reactions,such as nausea and vomiting,in group PCIA (2 cases)were more serious than that in the other groups (0 cases ).There were no significant differences among the other groups.Conclusion Ropivacaine plays an effective role in infiltration an-algesia when its concentration reaches 0.3% subcutaneous after radical mastectomy.

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