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Objective:To evaluate the role of 1, 4, 5-inositol triphosphate receptor (IP3R) in necroptosis of hippocampal neurons induced by sevoflurane anesthesia in aged rats.Methods:Sixty healthy male Sprague-Dawley rats, aged 18 months, weighing 500-600 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S) and sevoflurane anesthesia + IP3R antagonist group (group S+ I). S and S+ I groups inhaled 2% sevoflurane for 5 h. In group S+ I, IP3 receptor antagonist 2-APB 3 mg/kg was intraperitoneally injected at 10 min before sevoflurane inhalation, and the equal volume of dimethyl sulfoxide was intraperitoneally injected in group C and group S. Morris water maze test was used to test the cognitive function on the day after the end of sevoflurane anesthesia.Then the animals were sacrificed and the brain tissues were obtained for microscopic examination of the pathological changes after HE staining and Nissl staining (with a light microscope) and for determination of the free calcium concentration ([Ca 2+ ] i) and rate of necroptosis of hippocampal neurons (by flow cytometry) and expression of IP3R, receptor-interacting protein kinase-1 (RIPK1), receptor-interacting protein kinase-3 (RIPK3) and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, the times of crossing the platform were reduced, the time of staying at the target quadrant was shortened, the [Ca 2+ ] i and necroptosis rate of hippocampal neurons were increased, and the expression of IP3R, RIPK1, RIPK3 and p-MLKL in hippocampal neurons was up-regulated in group S and group S+ I ( P<0.05). Compared with group S, the escape latency was significantly shortened, the times of crossing the platform were increased, the time of staying at the target quadrant was prolonged, the [Ca 2+ ] i and necroptosis rate of hippocampal neurons were decreased, and the expression of IP3R, RIPK1, RIPK3 and p-MLKL in hippocampal neurons was down-regulated in group S+ I ( P<0.05). Conclusions:The mechanism by which sevoflurane induces cognitive dysfunction may be related to the imbalance of calcium homeostasis caused by activation of IP3R and thus inducing programmed necrosis in aged rats.
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Objective:To investigate the characteristics of an abnormal attachment point of the anterior horn of the medial meniscus (AHMM) by methods of arthroscopy, radiological imaging, and pathology.Methods:A total of seven cases of a rare abnormal attachment (anteromedial meniscofemoral ligament, AML) connecting the AHMM and anterior cruciate ligament (ACL) were identified in the more than six thousand arthroscopic procedures from January 2018 to April 2021. During the procedures, subtotal meniscal resection or meniscal repair was performed in all cases. Tissue removed intraoperatively was used for pathologic examination. For this study, the preoperative MRI films of these patients were retrospectively examined.Results:Finally, seven cases with diagnosed meniscal tears were included in this study, including 4 males and 3 females with a mean age of 37.85±7.70 years (range, 27-50 years). The AML, which extends outward and upward and is anterior to the ACL, begins at the AHMM and ends anterior to the femoral attachment of the ACL. The histological staining showed that the AML was a bundle of collagen fibers, whereas the ACL was a bundle of dense fibrous collagen. The AML could be clearly identified in the proton-weighted image of sagittal MRI preoperatively and showed a low signaling similar to the signal intensity of the ACL. AML was easily identified on sagittal MRI but not on axial and coronal MRI. In three patients who underwent bilateral MRI, the AML structure was found in both knees.Conclusion:The incidence of rare anatomic variation between the AHMM and the lateral condyle of the femur is 0.1%, usually occurring in both knees simultaneously. In cases with this variation, there is no bony attachment in the AHMM connected to the ACL via the AML. After transection of the AML, the annular fibrous structure and mechanical properties of the meniscus are damaged, as consequently the AML should be carefully protected in clinical practice.
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Objective:To compare the clinical outcome between arthroscopically-assisted modified outside-in technique and inside-out technique for reconstruction of posterior cruciate ligament (PCL).Methods:A retrospective cohort study was conducted to analyze the clinical data of 53 patients with PCL rupture treated at Zhengzhou Orthopaedics Hospital from September 2015 to March 2019. There were 36 males and 17 females, at age of 21-58 years [(37.2±12.5)years]. There were 29 patients with right knee injury and 24 patients with left knee injury. All patients underwent minimally invasive arthroscopically- assisted PCL reconstruction, including the femoral tunnel established using the modified outside-in technique in 22 patients (outside-in group) and the inside-out technique with 120° of knee flexion in 31 patients (inside-out group). The two groups were compared in terms of operation time, length of femoral tunnel intraoperatively, angle between femoral tunnel and intra-articular graft at postoperative 3 days, and posterior drawer test (PDT), International Knee Documentation Committee (IKDC) score, Lysholm knee score, knee range of motion (ROM), knee extension muscle strength at the last follow-up. The complications were recorded after operation.Results:All patients were followed up for 15-38 months [(21.4±8.7) months]. There were no significant differences between the two groups in operation time and last follow-up evaluation of PDT, IKDC score, Lysholm knee score and knee ROM (all P>0.05). The length of femoral tunnel in outside-in group [(38.6±2.9)mm] was longer than that in inside-out group [(32.4±2.5)mm] ( P<0.05). The angle between femoral tunnel and intra-articular graft in outside-in group [(147.5±3.1)°] was larger than that in inside-out group [(136.4±2.6)°] ( P<0.01). The knee extension muscle strength of all patients reached grade V at the last follow-up. There were 2 patients with 5° flexion limitation in outside-in group, comparable to 3 patients with 5° flexion limitation and 1 patient with 10° flexion limitation in inside-out group ( P>0.05). No incision infection, implant loosening or injury of the extension apparatus of the knee occurred after operation. Conclusions:In treating PCL rupture, both arthroscopically-assisted modified outside-in technique and inside-out technique can achieve satisfied stability and functional recovery. However, the length and angle of femoral tunnel is more controllable when using the the modified outside-in technique.
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Objective:To investigate the effect of activation of microglia in prefrontal cortex on long-term spatial memory in post-stroke depression mice.Methods:Forty-eight male C57BL/6 mice were divided into sham operation group, stroke group, post-stroke depression group and depression group according to the random number table method with 12 in each group, and 36 mice were divided into solvent group, enrofloxacin group and minocycline group according to the random number table method with 12 in each group.Middle cerebral artery occlusion (MCAO) was use to establish the stroke model, and forced swimming was used to establish the depression model.The post-stroke depression model mice were received MCAO first and then received forced swimming on the 4th day after stroke to establish the model.Mice in enrofloxacin group and minocycline group were treated with enrofloxacin and minocycline injection once a clay for 14 days from the 5th day after stroke, respectively.Forced swimming test and sugar water preference test were used to evaluate the depression of mice in each group, Morris water maze test was used to detect the spatial memory function of mice in each group, and Nissl staining and immunofluorescence staining were used to detect the neuronal function and the number and type of microglia activation.The expression of inflammatory cytokines IL-6 and IL-1β were detected by Western blot.GraphPad Prism 8.0.1 statistical software was used for statistical analysis.The single factor variance analysis was used to compare the difference among multiple groups, and pairwise comparison was performed with SNK- q test. Results:(1) There were statistically significant differences in depression, learning and memory, neuron damage, activation of microglia, inflammatory factors and other indicators in sham operation group, stroke group, post-stroke depression group and depression group ( F=43.58-255.70, all P<0.05). Compared with stroke group, post-stroke depression group had longer floating immobility time ((222.70±29.12) s, (79.25±46.78) s, P<0.05), the preference rate of sugar water was significantly lower ( (49.44±6.19) %, (84.49±4.73) %, P<0.05), and the average value of platform approach after correction was higher((125.00±9.95) mm, (96.79±12.57) mm, P<0.05), Nissl bodies expression was lower ((53.50±15.78) cells /mm 2, (85.67±17.52) cells /mm 2, P<0.05), NeuN positive expression rate was lower ((29.78±3.70) %, (45.73±4.51) %, P<0.05), the percent of M1 microglia expression was significantly higher ((75.55±8.84) %, (58.19±5.69) %, P<0.05), the percent of M2 microglia expression was lower ((43.46±5.11)%, (57.14±5.40)%, P<0.05), and the expression levels of IL-6 ((1.14±0.03), (0.94±0.05), P<0.05) and IL-1β((1.17±0.03), (0.56±0.04), P<0.05) were significantly higher.(2) Depression, learning and memory, neuron injury, activation of microglia, inflammatory factors and other indicators of mice in solvent group, enrofloxacin group and minocycline group were significantly different ( F=7.13-94.35, all P<0.05). Compared with enrofloxacin group, mice in minocycline group had shorter floating immobility time ((169.30±13.04) s, (224.30±22.60) s, P<0.05) and higher sugar water preference rate ((62.81±7.75) %, (47.71±8.11) %, P<0.05), the mean value of platform approach estimation after water maze correction was lower ((97.66±14.56) mm, (120.20±12.08) mm, P<0.05), and the expression level of Nissl bodies was higher ((80.17±10.55) cells /mm 2, (52.00±8.94) cells /mm 2, P<0.05), NeuN expression rate was high ((45.04±3.62) %, (28.88±4.50) %, P<0.05), Iba-1 expression was lower ((97.33±10.67) cells/mm 2, (112.50±6.54)cells/mm 2, P<0.05), the percent of M1 microglia expression was lower ((54.43±5.22) %, (73.82±6.88) %, P<0.05), and the percent of M2 microglia expression was significantly higher ((51.86±6.22) %, (36.30±5.72) %, P<0.05). The expression levels of IL-6 ((0.75±0.06), (1.21±0.07), P<0.05) and IL-1β ((0.61±0.06) (1.09±0.09), P<0.05) were lower. Conclusion:The long-term spatial memory impairment of post-stroke depression mice is aggravated, which is related to the neuron damage caused by increased activation of M1 microglia in PFC area.Inhibition of M1 microglia by minocycline can effectively improve the spatial memory ability of mice.
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Objective:To evaluate the effect of carbon monoxide-releasing molecule-3 (CORM-3) on blood transfusion-related acute lung injury in rats with traumatic brain injury (TBI).Methods:Seventy-two clean-grade healthy adult male Sprague-Dawley rats, weighing 300-350 g, were divided into 4 groups ( n=18 each) using the random number table method: sham operation group (group S), TBI group (T group), TBI plus 10 ml/kg plasma transfusion group (TP group), and TBI plus 10 ml/kg plasma transfusion plus CORM-3 group (TPC group). TBI was induced by dropping a 20-g weight from 20 cm height falling freely in anesthetized rats.Plasma 10 ml/kg was infused via the femoral vein after TBI in TP and TPC groups.The rats were sacrificed at 24 h after plasma transfusion, and lung tissues were obtained for determination of wet/dry weight (W/D) ratio, cell apoptosis, and expression of caspase-3, Bid, Bim and Puma (by Western blot). The lung injury score was calculated using the results of HE staining.Lung ultrasonography was performed for assessment of sonographic score, and the apoptosis rate was calculated by the TUNEL staining method. Results:Compared with S group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in the other three groups ( P<0.05). Compared with T group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in TP group ( P<0.05). Compared with TP group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly decreased, and the expression of activated caspase-3, Bid, Bim and Puma was down-regulated in TPC group ( P<0.05). Conclusion:CORM-3 can reduce acute lung injury related to blood transfusion in rats with TBI, and the mechanism may be related to inhibiting cell apoptosis in lung tissues.
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Objective:To evaluate the effect of VX-765 on cognitive function in acute rapid eye movement (REM) sleep-deprived juvenile rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 3-4 weeks, weighing 52-101 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), acute REM group (group AREM) and VX-765 group (group V). Sleep deprivation model was established by modified multi-platform water environment method.In group V, VX-765 solution 10 mg/kg was intravenously injected via the tail vein at 9: 00 a. m.every day for 4 consecutive days.The equal volume of normal saline was given instead in C and AREM groups.Morris water maze and novel object recognition tests were performed for 4 consecutive days during sleep deprivation.The rats were then sacrificed after the end of Morris water maze and novel object recognition tests on 5th day, and hippocampi were removed for determination of the expression of interleukin-1beta (IL-1β) and IL-18 by Western blot. Results:Compared with group C, the latency of novel object recognition was significantly prolonged, the percentage of novel object exploration was shortened, and the number of head exploration was decreased, the percentage of novel object exploration and discrimination index were decreased, the number of crossing the original platform in Morris water maze test was reduced, the time of staying at the target quadrant was shortened, and the expression of IL-1β and IL-18 was up-regulated in AREM and V groups ( P<0.05). Compared with group AREME, the latency of novel object recognition was significantly shortened, the percentage of novel object exploration was prolonged, and the number of head exploration was increased, the percentage of novel object exploration and discrimination index were increased, the number of crossing the original platform in Morris water maze test was increased, the time of staying at the target quadrant was prolonged, and the expression of IL-1β and IL-18 was down-regulated ( P<0.05). Conclusion:VX-765 can improve the cognitive function in acute REM sleep-deprived juvenile rats, which is related to inhibiting hippocampal inflammatory responses.
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Objective:To evaluate the effect of exogenous carbon monoxide (CO) on cell apoptosis during acute renal injury induced by hemorrhagic shock and resuscitation (HSR) in rats.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 4 groups ( n=12 each) by a random number table method: sham operation group (S group), HSR group (H group), HSR plus CORM-3 group (HC group) and HSR plus iCORM-3 group (HiC group). Mean arterial pressure was maintained at 30-35 mmHg for 45 min by withdrawing blood from the femoral vein, and the shed blood was re-transfused within 15 min to reach the initial blood pressure for resuscitation.Normal saline was infused when necessary, and the model of HSR was established.CORM-3 4 mg/kg and iCORM-3 4 mg/kg were added during resuscitation in HC group and HiC group, respectively.Only femoral vein and artery puncture was performed in S group.Blood samples were obtained from the tail vein at 3 h after resuscitation for measurement of serum urea nitrogen (BUN) and creatinine (Scr) concentrations.Rats were sacrificed at 12 h after resuscitation, and renal tissues were obtained for determination of the expression of Bcl-2 and Bak protein and cleaved caspase-3 (by Western blot) and cell apoptosis (by TUNEL). The damage to the renal tubules was assessed by paller assay after HE staining.Bcl-2/Bak ratio and apoptosis rate were calculated. Results:Compared with group S, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly increased, Bcl-2/Bak ratio was decreased, and the expression of cleaved caspase-3 was up-regulated in H, HC and HiC groups ( P<0.05). Compared with group H, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly decreased, Bcl-2/Bak ratio was increased, and the expression of cleaved caspase-3 was down-regulated in group HC ( P<0.05). Compared with group HC, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly increased, Bcl-2/Bak ratio was decreased, and the expression of cleaved caspase-3 was up-regulated in group HiC ( P<0.05). There was no significant difference in the indexes mentioned above between group H and group HiC ( P>0.05). Conclusion:The mechanism by which exogenous CO improves acute kidney injury may be related to inhibiting cell apoptosis in a rat model of HSR.
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Objective:To evaluate the effect of selective inhibitor of caspase-1 VX-765 on cognitive function in a rat model of hemorrhage shock and resuscitation (HSR).Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (S group), HSR group (H group), VX-765 group (V group), and solvent control group (C group). The rats in H, V and C groups were subjected to hemorrhage by bleeding from femoral vein to achieve mean arterial pressure of 25-35 mmHg which was maintained at this level for 60 min followed by resuscitation with shed blood within 15 min to restore blood pressure, and normal saline was infused when needed.VX-765 1 mg/kg and 0.4% polyethylene glycol 1 mg/kg were intravenously injected via the femoral vein immediately after the end of resuscitation in V and C groups, respectively.Six rats in each group were selected and sacrificed at 12 h after the end of resuscitation, and the cerebral cortex was removed for determination of neuronal pyroptosis (by immunofluorescence) and degree of cortical edema (using T2-weighted imaging). Cognitive function was measured by open field test on day 7 after resuscitation in the rest 6 rats in each group. Results:Compared with S group, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly increased, the distance in the central square and the number of standing on the back legs were decreased on day 7 after resuscitation, and the time spent in the central square was shortened in H, V and C groups ( P<0.05). Compared with H and C groups, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly decreased, the distance in the central square and the number of standing on the back legs were increased on day 7 after resuscitation, and the time spent in the central square was prolonged in V group ( P<0.05). Conclusion:VX-765 can improve the cognitive function, and the mechanism may be associated with inhibiting pyroptosis in cortical neurons in a rat model of HSR.
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Objective To evaluate the effect of carbon monoxide (CO) postconditioning on pyroptosis induced by oxygen-glucose deprivation and restoration (OGD/R) in rat hippocampai neurons and the relationship with mitochondrial permeability transition pore (mPTP)/reactive oxygen species (ROS) signaling pathway.Methods Primary hippocampal neurons were cultured in vitro,seed in 6-well or 96-well plates,and divided into 5 groups (n =24 each) using a random number table method:control group (C group),OGD/R group,CO postconditioning group (CO group),specific mPTP opener atractyloside plus CO postconditioning group (ACO group),and specific ROS inducer antimycin A plus CO postconditioning group (KCO group).Neurons were subjected to O2-glucose deprivation (OGD) for 16 h followed by restoration of O2-glucose supply for 24 h to establish the model of OGD/R injury.In group CO,neurons were exposed to 2％ CO-5％ CO2 for 3 h at 37 ℃ starting from the end of OGD,followed by normal culture for 21 h.In ACO and KCO groups,atractyloside 20 μmol/L and antimycin A 50 μmol/L were added at the end of OGD,respectively,and the other treatments were similar to those previously described in group CO.Neuronal pyroptosis rate was determined using double immunofluorescent staining cleaved caspase-1-AlexaFluor 568/DAPI after the end of treatments in each group.The neuronal survival rate was determined by MTT,opening of mPTP by Calcein-AM fluorescence,ROS content by DCFH-DA,and expression of interleukin1beta (IL-1β) and IL-18 by Western blot.Results Compared with C group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in the other groups (P＜0.05).Compared with OGD/R group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly decreased,the neuronal survival rate was increased,and the expression of IL-1β and IL-18 was down-regulated in CO,ACO and KCO groups (P＜0.05).Compared with CO group,neuronal pyroptosis rate and ROS content were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in ACO and KCO groups,and opening of mPTP was significantly inctreased in ACO group (P＜0.05).Conclusion CO postconditioning can inhibit OGD/R-induced pyroptosis in rat hippocampal neurons,and the mechanism is related to inhibiting mPTP/ROS signaling pathway.
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Objective To discuss the clinical effect of surgical treatment for ankle osteoarthritis under arthroscopy.Methods Forty-eight cases with ankle osteoarthritis patients admitted at the Orthopaedic Hospital of Zhengzhou from January 2012 to June 2014 were selected and all of them were given the treatment of focal cleaning under arthroscopy.The clinical effect of surgical treatment were judged by modified McGuire ankle rating system,the United States after ankle surgery AOFAS ankle-full score and Mazur ankle rating system respectively.Results At the time of the last follow-up,modified McGuire ankle rating system((85.64±16.52)points vs.(52.46±10.25)points,t=-8.465),the United States after ankle surgery AOFAS ankle-full score [(85.24±11.46)points vs.(53.68±9.48)points,t=-7.548)and Mazur ankle rating system((86.45±12.57)points vs.(58.49±8.64)points,t=-6.596)all increased than that of pre-operation,the differences were statistically significant(P＜0.05).The modified McGuire ankle rating system of patients with low-grade lesion at pre-operation((62.45±7.63)points vs.(49.58±6.35)points,t=3.685)and the time of the last follow-up((93.68±11.54)points vs.(68.54±9.68)points,t=8.695)were all higher than that of patients with high-grade lesion,the differences were statistically significant(P＜0.05).The clinical effect of surgical treatment judged by modified McGuire ankle rating system,the United States after ankle surgery AOFAS ankle-full score and Mazur ankle rating system were respectively 91.67％(44/48),89.58％(43/48)and 89.58％(43/48),the differences were no statistically significant(x2=0.824,P＞0.05).Conclusion The clinical effect of surgical treatment for ankle osteoarthritis under arthroscopy is remarkable and it causes light damage to the body.It is especially suitable for patients with low-grade lesions and is worth popularization and application.