Objective:To evaluate the effect of esketamine on cerebral ischemia-reperfusion (I/R) injury and the association with mitochondrial stress in mice.Methods:The experiment was performed in two parts. Part Ⅰ Eighteen SPF male C57BL/6 mice, aged 8-12 weeks, with body mass index of 28-30 g, were divided into 3 groups ( n=6 each) by a random number table method: sham operation group (S group), cerebral I/R group (IR group), and esketamine plus cerebral I/R group (E+ IR group). Cerebral I/R was produced by occlusion of middle cerebral artery for 1 h followed by 24-h reperfusion in anesthetized mice.Esketamine 10 mg/kg was intraperitoneally injected at 20 min before developing the model in E group. Neurological function was evaluated using the Zea Longa score and balance beam test (Feeney score). The cerebral infarct size was determined by TTC staining. Part Ⅱ Primary cortical neurons were isolated and cultured and then divided into 3 groups ( n=42 each) using a random number table method: control group (group C), oxygen-glucose deprivation-reoxygenation (OGD/R) group, and esketamine plus OGD/R group (group E+ OGD/R). Cells were subjected to O 2-glucose deprivation for 1 h followed by restoration of O 2-glucose supply for 24 h. The cells were treated with 25 μmol/L esketamine for 40 min before preparing the model in E+ OGD/R group. The neuronal viability was measured by the CCK-8 assay. The ultrastructure of neurons was observed with a transmission electron microscope. The levels of reactive oxygen species (ROS), glutathione peroxidase (GSH-px) and malondialdehyde (MDA) were determined, and the mitochondrial membrane potential was determined by JC-1 kit. The neuronal apoptosis was detected by TUNEL staining, and the apoptosis rate of neurons was calculated. The expression of Bax, cytochrome C (CytC), cleaved-caspase-9, caspase-3 and cleaved-caspase-3 was detected by Western blot. Results:Part Ⅰ Compared with S group, the Zea Longa score, Feeney score and cerebral infarct size were significantly increased in IR group ( P<0.01). Compared with IR group, the Zea Longa score, Feeney score and cerebral infarct size were significantly decreased in E+ IR group ( P<0.01). Part Ⅱ Compared with C group, the cell viability and activity of GSH-px were significantly decreased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were increased, and the expression of Bax, Cyt C and cleaved-caspase-9 was up-regulated in OGD/R group ( P<0.01). Compared with OGD/R group, the cell viability and activity of GSH-px were significantly increased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were decreased, and the expression of Bax, Cyt C and cleaved-caspase-9 was down-regulated in E+ OGD/R group ( P<0.01). Conclusions:Esketamine can alleviate cerebral I/R injury in mice, and the mechanism may be related to inhibition of mitochondrial stress in neurons, improvement in mitochondrial function, and inhibition of mitochondria-dependent apoptosis in neurons.

Objective:To study the effects of different single cell phenotypes on the prognosis of patients with intrahepatic cholangiocarcinoma by using spatial analysis, providing clues for obtaining potential immunotherapeutic targets.Methods:This study was a retrospective cohort study. A total of 41 intrahepatic cholangiocarcinoma (ICC) patients who underwent surgery in Beijing Youan Hospital Affiliated to Capital Medical University from February 2013 to June 2019 were enrolled. According to the 5-year survival situation, the patients were divided into survival group ( n=10) and death group ( n=31). A metal label-based tissue imaging mass panel containing 36 related markers was designed and constructed for staining different components in tumor samples. Through the analysis of the type and quantity of different metacluster and spatial location information and combined with the clinical outcomes of patients with information, certain metaclusters were found related to the prognosis of patients. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), paired t-test was used for comparison between groups. Measurement data with skewed distribution were expressed as median, and rank sum test was used for comparison between groups. The chi-square test was used to compare the counting data between groups. Results:36 biomarkers of 41 ICC patients were located and quantified to generate 1 476 single-cell resolution histological images. The expression information of various markers was analyzed by t-distributed Stochastic Neighbor Embedding (tSNE), and subgroups annotations (1-29) were added. It revealed that the density of metacluster 7(CD8 + T cells) was lower in survival group. The density of metacluster 16(Bcl-2 + CK7 + cancer cells) within tumors, as well as the density of metacluster 3(Vista + GB + CD11b + neutrophils) within stroma were higher in death group. Conclusion:The density of metacluster 7(Activated CD8 + T cells), metacluster 16(Bcl-2 + CK7 + tumor cells) and a novel neutrophil metacluster 3(Vista + GB + CD11b + neutrophils) correlated with ICC patients prognosis.

Objective:To investigate the application value of Laennec approach in laparoscopic anatomical right hemihepatectomy (LARH).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 2 female patients who underwent LARH via Laennec approach in the First Affiliated Hospital of Kangda College of Nanjing Medical University from May to July 2020 were collected. The two patients were 51 and 57 years old, respectively. Observation indicators: (1) surgical situations; (2) postoperative situations and follow-up. Follow-up using outpatient examination and telephone interview was conducted to detect post-operative survival and tumor recurrence of patients up to December 2020. Count data were repre-sented as absolute numbers.Results:(1) Surgical situations: 2 patients successfully underwent LARH via Laennec approach, without conversion to open surgery. The operation time was 180 minutes and 185 minutes, and the volume of intraoperative blood loss was 200 mL and 400 mL, respectively. No blood transfusion or gastrointestinal decompression was performed in either patient. (2) Postoperative situations and follow-up: 2 patients began to take liquid diet on the first day and out-of-bed activities on the postoperative second to third day. There was no postoperative bile fistula or bleeding, but different degrees of peritoneal and pleural effusion occurred to the 2 patients after operation. One case was improved after right-sided thoracentesis and chest tube drainage due to dyspnea, and the other case was cured after conservative therapy. There was no perioperative death. The duration of postoperative hospital stay of 2 patients was 13 days and 11 days, respectively. Results of pathological examination showed 1 case of hepatic hemangioma and 1 case of primary liver cancer, respectively. The Laennec capsule was observed on the hepatic vein branches of segment Ⅴ, Ⅵ, Ⅶ, Ⅷ, and the gap existed between the Laennec capsule and the hepatic vein. Two patients were followed up for 7 months and 5 months，respectively. They survived during the follow-up，without tumor recurrence.Conclusion:It is safe and feasible to perform LARH by Laennec approach.

BACKGROUND: It is a great challenge for surgeons to resect pulmonary nodules with small volume, deep position and no solid components under video-assisted thoracoscopic surgery. The purpose of this study is to explore the feasibility and necessity of the localization of pulmonary nodules by injecting indocyanine green (ICG) under the guidance of magnetic navigation bronchoscope and the resection of small pulmonary nodules under the fluoroscope. METHODS: Between December 2018 and August 2019, sixteen consecutive patients with 30 peripheral lung lesions in our hospital received fluorescent thoracoscopic pulmonary nodule resection. Electromagnetic navigation bronchoscope (ENB) was performed before surgery to guide ICG to the target lesion. RESULTS: All patients underwent magnetic navigation-guided pulmonary nodule localization, and surgical resection was performed immediately after localization was completed. The average size of the nodules was (11.12±3.65) mm. The average navigation time was (12.06±2.74) minutes, and the average interval between dye labeling and lung resection was (25.00±5.29) minutes. All lesions were completely resected, the localization success rate was 100.00%, no bleeding and other complications occurred after the localization, the postoperative pathological results confirmed the accuracy of the staining. CONCLUSIONS Indocyanine green injection under the guidance of magnetic navigation bronchoscope is an effective way to locate pulmonary nodules, which can locate small and untouchable lesions in the lung. This method can help surgeons identify lesions more quickly and accurately. It is practical and worthy of promotion.

Objective To evaluate the role of 2B subunit-containing NMDA receptors ( NR2B) in the rostal anterior cingulate cortex ( rACC) in development of pain-related aversion in a rat model of bone cancer pain using siRNA technique. Methods Forty-five healthy male Wistar rats, weighing 220-250 g, were divided into 3 groups ( n=15 each) using a random number table method: normal saline blank control group ( group NS) , NR2B-siRNA lentivirus group ( group LV-NR2B) and pGC-FU-siRNA lentivirus group (group LV-NC). A total volume of MADB-106 cells 3μl (4. 8×109 cells∕ml) was inoculated into the bone marrow cavity of the right tibia of rats. At day 2 after inoculation, NR2B∕siRNA recombinant lentivirus 0. 2μl was injected into rACC in group LV-NR2B, and pGC-FU-siRNA negative recombinant lentivirus 0. 2μl was injected into rACC in group LV-NC. The mechanical paw withdrawal threshold ( MWT) was measured at 1 day before inoculation and 3, 7, 14 and 21 days after inoculation. Conditioned place avoidance test was performed at 1 day before inoculation and 14 days after inoculation, and the percentage of residence time in room A was calculated. The rats were sacrificed at 21 days after inoculation and the rACC was re-moved for detecting NR2B protein and mRNA expression by Western blot or real-time polymerase chain re-action. Results Compared with the baseline at 1 day before inoculation, the MWT was significantly de-creased at 7, 14 and 21 days after inoculation, and the percentage of residence time in room A was de-creased at 14 days after inoculation in NS and LV-NC groups (P<0. 05), and no significant change was found in the parameters mentioned above in LV-NR2B group (P>0. 05). Compared with group NS, the MWT was significantly increased at 14 and 21 days after inoculation, the percentage of residence time in room A was increased at 14 days after inoculation, and the expression of NR2B protein and mRNA was down-regulated in group LV-NR2B ( P<0. 05) , and no significant change was found in the parameters men-tioned above in group LV-NC (P>0. 05). Conclusion Up-regulated expression of NR2B in rACC is in-volved in development of pain-related aversion in a rat model of bone cancer pain.

Dexmedetomidine is an α2-adrenergic receptor agonist that exhibits a protective effect on ischemia-reperfusion injury of the heart, kidney, and other organs. In the present study, we examined the neuroprotective action and potential mechanisms of dexmedetomidine against ischemia-reperfusion induced cerebral injury. Transient focal cerebral ischemia-reperfusion injury was induced in Sprague-Dawley rats by middle cerebral artery occlusion. After the ischemic insult, animals then received intravenous dexmedetomidine of 1 μg/kg load dose, followed by 0.05 μg/kg/min infusion for 2 h. After 24 h of reperfusion, neurological function, brain edema, and the morphology of the hippocampal CA1 region were evaluated. The levels and mRNA expressions of interleukin-1β, interleukin-6 and tumor nevrosis factor-α as well as the protein expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κBp65, inhibitor of κBα and phosphorylated of κBα in hippocampus were assessed. We found that dexmedetomidine reduced focal cerebral ischemia-reperfusion injury in rats by inhibiting the expression and release of inflammatory cytokines and mediators. Inhibition of the nuclear factor-κB pathway may be a mechanism underlying the neuroprotective action of dexmedetomidine against focal cerebral I/R injury.
Animals ; Brain Edema ; CA1 Region, Hippocampal ; Cyclooxygenase 2 ; Cytokines ; Dexmedetomidine* ; Heart ; Hippocampus ; Infarction, Middle Cerebral Artery ; Inflammation* ; Interleukin-6 ; Kidney ; Neuroprotection* ; Nitric Oxide Synthase Type II ; Rats* ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury* ; RNA, Messenger

Dexmedetomidine is an α2-adrenergic receptor agonist that exhibits a protective effect on ischemia-reperfusion injury of the heart, kidney, and other organs. In the present study, we examined the neuroprotective action and potential mechanisms of dexmedetomidine against ischemia-reperfusion induced cerebral injury. Transient focal cerebral ischemia-reperfusion injury was induced in Sprague-Dawley rats by middle cerebral artery occlusion. After the ischemic insult, animals then received intravenous dexmedetomidine of 1 μg/kg load dose, followed by 0.05 μg/kg/min infusion for 2 h. After 24 h of reperfusion, neurological function, brain edema, and the morphology of the hippocampal CA1 region were evaluated. The levels and mRNA expressions of interleukin-1β, interleukin-6 and tumor nevrosis factor-α as well as the protein expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κBp65, inhibitor of κBα and phosphorylated of κBα in hippocampus were assessed. We found that dexmedetomidine reduced focal cerebral ischemia-reperfusion injury in rats by inhibiting the expression and release of inflammatory cytokines and mediators. Inhibition of the nuclear factor-κB pathway may be a mechanism underlying the neuroprotective action of dexmedetomidine against focal cerebral I/R injury.
Animals ; Brain Edema ; CA1 Region, Hippocampal ; Cyclooxygenase 2 ; Cytokines ; Dexmedetomidine* ; Heart ; Hippocampus ; Infarction, Middle Cerebral Artery ; Inflammation* ; Interleukin-6 ; Kidney ; Neuroprotection* ; Nitric Oxide Synthase Type II ; Rats* ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury* ; RNA, Messenger

Objective To evaluate the effect of dexmedetomidine on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty pathogen-free male Sprague-Dawley rats,aged 12-16 months,weighing 300-360 g,were randomly divided into 3 groups (n =10 each) using a random number table:sham operation group (group Sham),focal cerebral I/R group (group I/R),and dexmedetomidine group (group D).Focal cerebral I/R was induced by occlusion of the right middle cerebral artery.In group D,dexmedetomidine was given as a loading dose of 1 μg/kg (over 10 min) starting from 1 h of ischemia,followed by an infusion of 0.05 μg · kg-1 · h-1 until 2 h of reperfusion.Neurological deficit was assessed and scored at 24 h of reperfusion,and then the rats were sacrificed.Brains were removed for determination of cerebral infarct size and expression of iNOS and COX-2 in the hippocampus (by Western blot).The percentage of cerebral infarct size was calculated.Results Compared with group Sham,the neurological deficit score,percentage of head swing to the left,percentage of cerebral infarct size,and expression of iNOS and COX-2 in the hippocampus were significantly increased in I/R and D groups (P＜0.05).Compared with group I/R,the neurological deficit score,percentage of head swing to the left,percentage of cerebral infarct size,and expression of iNOS and COX-2 in the hippocampus were significantly decreased in group D (P＜0.05).Conclusion The mechanism by which dexmedetomidine mitigates focal cerebral I/R injury may be related to inhibition of iNOS and COX-2 expression in rats.

Objective To evaluate the effect of controlled hypotension at the beginning of reperfusion on ischemia-reperfusion (I/R) injury of the liver in patients undergoing hepatectomy.Methods Forty ASA Ⅱ or Ⅲ patients (aged 30-60 years and weighing 40-70 kg) undergoing elective partial hepatectomy for liver cancer were randomly divided into two groups (n =20 each):normal blood pressure group (control group,group C) and controlled hypotension group (group H).In group C,normal blood pressure was maintained during reperfusion,while in group H,controlled hypotension (the mean arterial blood pressure (MAP) was maintained at 60-70 mm Hg) was performed for 10 minutes since the beginning of reperfusion.Hepatic portal was occluded during operation.Venous blood samples were taken before hepatic ischemia (T0,baseline) and after 15 minutes of ischemia (T1) and after 25 minutes of reperfusion (T2) for determination of plasma levels of endothelin (ET),nitric oxide (NO),tumor necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1).Results I/R of the liver led to significant increases in plasma levels of ET,TNF-α and IL-1 and a decrease in plasma level of NO at T1,2 as compared with the baseline values at T0 in both groups.Plasma levels of ET,TNF-α and IL-1 were significantly lower while plasma level of NO was significantly higher at T2 in group H than in group C.Conclusion Controlled hypotension for 10 minutes in the initial stage of reperfusion can attenuate I/R-induced injury to the liver in patients undergoing hepatectomy through balancing ET with NO and inhibiting inflammation responses.

Objective To investigate the effect of dexmedetomidine on postoperative patient-controlled intravenous analgesia (PCIA) with sufentanil in patients with essential hypertension. Methods Sixty ASA Ⅱ or Ⅲ patients with essential hypertension aged 42-63 yr weighing 48-72 kg undergoing hysterectomy were randomly divided into 3 groups ( n = 20 each): control group ( group C) and different doses of dexrmedetomidine groups ( group D1.2 ). PCIA was performed with sufentanil 1 μg/ml + tropisetron 5 μg/ml in 100 ml of normal saline within 24 h after operation (background infusion at 2 ml/h with a bolus dose of 0.5 ml and a 15 min lockout interval). Dexmein group C. Ramsay score was recorded. The number of attempts, consumption of sufentanil, the number of patients who needed nifedipine or ephedrine and side effects such as vomiting and respiratory depression were recoded within 24 h after operation. The level of sedation was evaluated with Ramsay sedation score at 24 h after operation.Results Compared with group C, the number of attempts, consumption of sufentanil, the number of patients who needed nifedipine and incidences of vomiting and respiratory depression were significantly decreased, while Ramsay score was significantly increased in D1 and D2 groups, and the number of patients who needed ephedrine was significanlly increased in group D2 ( P ＜ 0.05). The number of attempts and consumption of sufentanil were significantly decreased, and Ramsay score and the number of patients who needed ephedrine were significantly increased in group D2 compared with group D1 ( P ＜ 0.05). Conclusion Dexmedetomidine can not only reduce the consumption of sufentanil for postoperative PCIA, but also prevent postoperative hypertension from deteriorating in patients with essential hypertension.