Objective:To evaluate the relationship between the neutrophil-to-lymphocyte ratio (NLR) during the perioperative period and early postoperative recurrence in patients with advanced gastric cancer.Methods:In this prospective study, 188 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-75 yr, with a body mass index of 20-30 kg/m 2, with radiological staging of T 2-4N xM 0, underwent elective gastric cancer radical surgery under general anesthesia at the Fourth Hospital of Hebei Medical University from January 2018 to April 2022, were selected. The NLR was measured at 1 day before operation and postoperative days 1, 3 and 5. The difference between postoperative and preoperative NLR (ΔNLR) was calculated. Based on the presence or absence of tumor recurrence and/or metastasis within 2 yr after surgery, the patients were divided into recurrence group and non-recurrence group. The Cox proportional hazards regression model was employed to evaluate the early postoperative recurrence and perioperative NLR in patients with advanced gastric cancer. Results:A total of 171 patients ultimately completed the follow-up. There were 77 cases in recurrence group and 94 in non-recurrence group. Compared to non-recurrence group, NLR and ΔNLR were significantly increased on the 5th day after operation in recurrence group ( P<0.05). The elevated NLR and ΔNLR at postoperative day 5 were independent risk factors for recurrence within 2 yr in patients with advanced gastric cancer ( HR=1.026, P<0.05). Conclusions:NLR may not be associated with the risk of early postoperative recurrence in the patients with advanced gastric cancer, and further evaluation with large-scale studies is needed.

Objective:To evaluate the relationship between NAD + -dependent protein deacylase sirtuin-1 (SIRT1) and tau protein during cardiopulmonary bypass (CPB)-induced neuroinflammation in rats. Method:Thirty-six healthy adult male Sprague-Dawley rats, weighing 250-300 g, were used in this study. Eighteen rats were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (C1 group), CPB1 group and tau protein inhibitor methionine+ CPB group (methionine+ CPB group). Another 18 rats were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (C2 group), CPB2 group and SIRT1 agonist SRT1720+ CPB group (SRT+ CPB group). Methionine+ CPB group received oral methionine 10.5 mg/kg once a day for 7 consecutive days. SRT1720 10 mg/kg was intraperitoneally injected once a day for 5 consecutive days in SRT+ CPB group. The model of CPB was developed after the end of drug administration. Rats were weaned off the bypass system after 1 h of circulatory support and sacrificed, and brain tissues were harvested for determination of the contents of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-4 (using enzyme-linked immunosorbent assay), expression of CD32, ARG1, SIRT1, tau protein and acetylated tau protein (ac-tau) (by Western blot), and expression of CD32, ARG1, SIRT1 and tau mRNA (by quantitative real-time polymerase chain reaction). Results:Compared with C1 group, the contents of TNF-α and IL-6 were significantly increased, the content of IL-4 was decreased, the expression of tau and CD32 protein and mRNA was up-regulated, and the expression of ARG1 protein and mRNA was down-regulated in CPB1 group ( P<0.05). Compared with CPB1 group, the contents of TNF-α and IL-6 were significantly decreased, the content of IL-4 was increased, the expression of tau and CD32 protein and mRNA was down-regulated, and the expression of ARG1 protein and mRNA was up-regulated in M+ CPB group ( P<0.05). Compared with C2 group, the contents of TNF-α and IL-6 were significantly increased, the content of IL-4 was decreased, the expression of ac-tau and tau and CD32 protein and mRNA was up-regulated, and the expression of SITR1 and ARG1 protein and mRNA was down-regulated in CPB2 group ( P<0.05). Compared with CPB2 group, the contents of TNF-α and IL-6 were significantly decreased, the content of IL-4 was increased, the expression of ac-tau and tau and CD32 protein and mRNA was down-regulated, and the expression of SITR1 and ARG1 protein and mRNA was up-regulated in SRT+ CPB group ( P<0.05). Conclusions:The down-regulated SIRT1 expression in brain tissues after cardiopulmonary bypass can inhibit deacetylation of tau protein, promote M1 polarization of microglia, inhibit M2 polarization, and ultimately induce neuroinflammation in rats.

Objective:To evaluate the relationship between postoperative delirium (POD) and concentrations of phosphatidylethanolamine-binding protein 1 (PEBP1) in cerebrospinal fluid (CSF) of elderly patients undergoing total knee and hip arthroplasty.Methods:In this case-control study, 375 elderly patients of both sexes, aged ≥65 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, scheduled for elective total knee and hip arthroplasty under combined spinal-epidural anesthesia at Haian Hospital Affiliated to Nantong University from November 2022 to June 2024, were selected. The perioperative clinical data were collected. CSF was drawn before anaesthesia for determination of the concentrations of PEBP1, Abeta 42 (Aβ 42), total tau (t-tau) and phosphorylated tau (p-tau) by enzyme-linked immunosorbent assay. The occurrence of postoperative delirium was evaluated using the Confusion Assessment Method at 1-7 days after operation. The severity of POD was assessed using a Memorial Delirium Assessment Scale. The patients were divided into POD group and non-POD group based on whether POD occurred. The influencing factors of POD were analyzed using multivariate logistic regression. Mediating effects were assessed to determine whether the levels of Aβ 42, p-tau and t-tau in CSF mediated PEBP1′s effect on POD. The accuracy of CSF PEBP1 concentration in predicting POD was evaluated using the receiver operating characteristic curve. Results:Decreased concentration of Aβ 42 in CSF, decreased ratios of Aβ 42/p-tau and Aβ 42/t-tau in CSF, and increased concentrations of PEBP1, p-tau and t-tau in CSF were independent risk factors for POD ( P<0.05). The results of mediation analysis showed that the relationship between PEBP1 and POD was partially mediated by CSF Aβ 42 (15.0%) and Aβ 42/t-tau ratio (14.9%). The receiver operating characteristic curve showed that the accuracy of CSF PEBP1 concentrations in predicting the occurrence of POD was high (AUC=0.846, P<0.001). Conclusions:Preoperative elevated CSF PEBP1 concentration is a risk factor for POD. CSF Aβ 42 concentration and Aβ 42/t-tau ratio serve as key mediators in the the association between PEBP1 and POD. PEBP1 concentration is more accurate in predicting POD.

Objective:To evaluate the role of inositol 1, 4, 5 triphosphate receptor 1 (IP3R1)-regulated changes in mitochondria-associated endoplasmic reticulum membrane (MAM) structure in the long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal mice.Methods:Sixty SPF-grade healthy neonatal C57BL/6J mice of either sex, aged 6 days, weighing 6-10 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), multiple sevoflurane anesthesia group (group S), and IP3R antagonist 2-APB+ multiple sevoflurane anesthesia group (group I+ S). Group S and group I+ S inhaled 3% sevoflurane anesthesia for 2 h starting from 6, 8 and 10 days after birth. In group I+ S, 2-APB 3 mg/kg was intraperitoneally injected before each sevoflurane anesthesia. The open field test was performed at day 31 after birth to assess the spontaneous mobility. The Morris water maze test was performed at days 31-36 after birth to assess the cognitive function. Mice were sacrificed at the end of the water maze test, hippocampal CA1 region was isolated and hippocampal tissues were obtained for determination of the intracellular calcium ion concentration ([Ca 2+ ] i) and rate of necroptosis (using Flow cytometry) and expression of IP3R1, G protein-coupled receptor 75 (GRP75), receptor-interacting protein kinase 1 (RIPK1), RIPK3, and phosphorylated human mixed-series protein kinase-like structural domains (p-MLKL) (by Western blot). Transmission electron microscopy was performed to observe and record the partial length of MAMs, endoplasmic reticulum circumference and mitochondrial circumference. Results:There were no statistically significant differences in the speed, distance, and time of staying at the center in open field tests among the three groups ( P>0.05). Compared with group C, the escape latency was significantly prolonged on postnatal days 33-35, the number of crossing the original platform was reduced, the necroptosis rate in the hippocampal CA1 region and [Ca 2+ ] i were increased, the expression of IP3R, GRP75, RIPK1, RIPK3 and p-MLKL was up-regulated, and the ratio of MAMs partial length/endoplasmic reticulum perimeter and ratio of MAMs partial length/mitochondria perimeter in hippocampal neurons were elevated in group S ( P<0.05). Compared with group S, the escape latency was significantly shortened on postnatal days 32-35, the number of crossing the original platform was increased, the necroptosis rate in the hippocampal CA1 region and [Ca 2+ ] i were decreased, the expression of IP3R, GRP75, RIPK1, RIPK3 and p-MLKL was down-regulated, and the ratio of MAMs partial length/endoplasmic reticulum perimeter and ratio of MAMs partial length/mitochondria perimeter in hippocampal neurons were decreased in group I+ S ( P<0.05). Conclusions:Structural changes in MAMs in the hippocampal CA1 region mediated by the up-regulation of IP3R1 expression are involved in the process of long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal mice.

Objective:To evaluate the efficacy of ultrasound-guided median interspinous in-plane approach to subarachnoid puncture in obese pregnant patients.Methods:This study was a randomized controlled trial. Eighty obese parturients who underwent elective cesarean section from March 2022 to January 2024 in our hospital were divided into 2 groups( n=40 each) by the random number table method: median interspinous in-plane approach group(group M) and paramedian interlaminar in-plane approach group(group P). After successful puncture, 0.5% ropivacaine 15 mg(3 ml) was intrathecally injected in both groups. The first-attempt success of puncture, the number of puncture attempts, operation time, the total success of puncture, and the visibility scores of the anterior and posterior union, positioning structure(lamina in group P, spinous process in group M) and puncture needle under ultrasound were recorded. Results:Compared with group P, the visibility score of positioning structure under ultrasound was significantly increased, the success rate of puncture at the first attempt was increased, the number of puncture attempts was decreased, the operation time was shortened, the total success rate of puncture was increased( P<0.05), and no significant change was found in the visibility scores of the anterior and posterior union and puncture needle in group M( P>0.05). Conclusions:For obese pregnant patients, the ultrasound-guided median interspinous in-plane approach can accurately and quickly perform subarachnoid puncture, which has more advantages than the traditional paramedian interlaminar in-plane approach.

Objective:To develop a nursing training program for malignant hyperthermia (MH) in perioperative patients based on the Instructional System Design model.Methods:Based on the Instructional System Design model, a preliminary draft of the nursing training program for MH was developed using literature review and semi-structured interviews in perioperative patients. Through expert consultations and the use of the Analytic Hierarchy Process, the content and weight of each item of the nursing training program for MH in perioperative patients were determined.Results:The effective response rates for the two rounds of expert consultation questionnaires were 84% and 97% respectively, with expert authority coefficients of 0.833 and 0.853 respectively, and Kendall′s harmony coefficients of 0.148 and 0.210 respectively ( P<0.01). The finalized nursing training program included 5 first-level items, 16 second-level items, and 61 third-level items. Conclusions:The construction process of the nursing training program for MH in perioperative patients is standardized and scientific, with detailed and practical content, which helps improve the level and quality of MH care.

Objective:To evaluate the effect of baicalein on acute myocardial injury in rats with high-level spinal cord injury (SCI) and the role of nuclear factor E2-related factor 2 (Nrf2).Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SCI group, SCI+ baicalein group (SCI+ Bai group) and SCI+ baicalein+ ML385 group (SCI+ Bai+ ML385 group). The high-level SCI rat model was established by the modified Allens method. In Sham group, the 7th cervical vertebra (C 7) was only exposed, but the spinal cord was not hit. In SCI group, C 7 was exposed and the spinal cord was hit. In SCI+ Bai group, baicalein 50 mg/kg was intraperitoneally injected immediately after SCI. In SCI+ Bai+ ML385 group, Nrf2 inhibitor ML385 30 mg/kg was intraperitoneally injected at 1 h before SCI, and baicalein 50 mg/kg was intraperitoneally injected immediately after SCI. The rats were anesthetized at 24 h after SCI and sacrificed after the blood samples from the abdominal aorta were collected and the hearts were taken for microscopic examination of the pathological changes (by HE staining) which were scored and the ultrastructure of cells (with a transmission electron microscope) and for determination of the serum cardiac troponin I (cTnI) concentrations (by enzyme-linked immunosorbent assay), content of ferrous ion (Fe 2+ ) in myocardial tissues (by colorimetry), contents of malondialdehyde(MDA) and glutathione (GSH) and activity of superoxide dismutase(SOD) in myocardial tissues (by biochemical method) and expression of glutathione peroxidase 4 (GPX4), acyl CoA synthase long chain family member 4 (ACSl4) and Nrf2 protein and mRNA in myocardial tissues (by Western blot and fluorescent quantitative polymerase chain reaction). The mitochondrial Flameng score was assessed and recorded. Results:Compared with Sham group, the pathological score, mitochondrial Flameng score and serum cTnI concentrations were significantly increased, the contents of Fe 2+ and MDA in myocardial tissues were increased, the content of GSH and SOD activity were decreased, the expression of GPX4 was down-regulated, and the expression of ACSL4 and Nrf2 was up-regulated in SCI group ( P<0.05). Compared with SCI group, the pathological score, mitochondrial Flameng score and serum cTnI concentration were significantly decreased, the contents of Fe 2+ and MDA in myocardial tissues were decreased, the contents of GSH and SOD activity were increased, the expression of GPX4 and Nrf2 was up-regulated, and the expression of ACSL4 was down-regulated in SCI+ Bai group ( P<0.05). Compared with SCI+ Bai group, the pathological score, mitochondrial Flameng score and serum cTnI concentrations were significantly increased, the contents of Fe 2+ and MDA in myocardial tissues were increased, the content of GSH and SOD activity were decreased, the expression of GPX4 and Nrf2 was down-regulated, and the expression of ACSL4 was up-regulated in SCI+ Bai+ ML385 group ( P<0.05). Conclusions:Baicalein can alleviate acute myocardial injury in rats with high-level SCI, and Nrf2 is involved in this process.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Objective:To evaluate the role of spinal Annexin A3 (ANXA3) in neuropathic pain in mice.Methods:Sixty-four SPF healthy adult male C57BL/6 mice, weighing 22-26 g, aged 8-10 weeks, were divided into 4 groups ( n=16 each) by the random number table method: sham operation group (group S), chronic constriction injury (CCI) group, CCI+ negative control adeno-associated virus AAV-NC group (group CCI+ N) and CCI+ adeno-associated virus AAV-shANXA3 group (group CCI+ sh). The neuropathic pain was induced by CCI of the sciatic nerve in anesthetized animals. The AAV-shANXA3 and AAV-NC (5 μl) were intrathecally injected at 14 days before developing the model in CCI+ N group and CCI+ sh group. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before developing the model and at 7, 14 and 21 days after developing the model. All the mice were sacrificed after the last measurement of pain threshold, the L 4-6 segments of the spinal cord were removed for determination of the expression of ANXA3, phosphorylated nuclear factor-kappa B (p-NF-κB) and ionized calcium-binding adaptor molecule-1 (Iba-1)(by Western blot), expression of ANXA3 mRNA (by real-time polymerase chain reaction), microglial activation (using the immunofluorescence staining), and contents of pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 and anti-inflammatory cytokines transforming growth factor-beta (TGF-β) and IL-10 (using enzyme-linked immunosorbent assay). Results:Compared with group S, the MWT was significantly decreased and TWL was shortened after developing the model, the expression of ANXA3 protein and mRNA, p-NF-κB and Iba-1 in spinal cord was up-regulated, the contents of TNF-α, IL-1β and IL-6 were increased, the contents of TGF-β and IL-10 were decreased ( P<0.05), the activation of microglia in the spinal cord was significantly increased, and the cell body was enlarged in group CCI. There was no significant difference in each parameter between group CCI and group CCI+ N ( P>0.05). Compared with CCI+ N group, the MWT was significantly increased on days 14 and 21 after developing the model, the TWL was prolonged on day 21 after developing the model, the expression of ANXA3 protein and mRNA, p-NF-κB and Iba-1 was down-regulated, the contents of TNF-α, IL-1β and IL-6 were decreased, the contents of TGF-β and IL-10 were increased ( P<0.05), and the activation of spinal microglia was decreased in CCI+ sh group. Conclusions:Spinal ANXA3 may be involved in the development and maintenance of neuropathic pain by activating the NF-κB signaling pathway and further promoting microglial activation in mice.

Objective:To evaluate the role of leucine-rich repeat kinase 2 (LRRK2) in the spinal dorsal horn in diabetic neuropathic pain (DNP) and to determine whether the mechanism involved nuclear factor kappa B (NF-κB) signaling pathway in rats.Methods:SPF healthy male Sprague-Dawley rats, aged 8 weeks, weighing 205-238 g, were fed a high-sugar and high-fat diet and received intraperitoneal injection of streptozotocin to induce a type 2 diabetes mellitus model. Successful DNP model was defined as the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) being below 85% of the baseline in type 2 diabetes mellitus rats. Thirty-six rats with DNP were allocated into 3 groups ( n=12 each) using a random number table method: DNP group, MLi-2 (LRRK2 inhibitor) group, and MLi-2+ phorbol 12-myristate 13-acetate (PMA) (NF-κB activator) group. MLi-2 1 mg/kg was intrathecally injected at L 5-6 in MLi-2 group. In MLi-2+ PMA group, MLi-2 1 mg/kg was intrathecally injected, and 1 h later PMA 20 μg/10 μl was intrathecally injected. In group DNP, the equal volume of 5% dimethyl sulfoxide was intrathecally injected. Twelve healthy rats were randomly selected and served as control group (C group). The animals were fed a standard diet, normal saline 2 ml was intraperitoneally injected, and the equal volume of 5% dimethyl sulfoxide was intrathecally injected in group C. Intrathecal injection was performed once a day for 7 consecutive days. The MWT and TWL were measured at 1, 3, 5 and 7 days after intrathecal injection, and the motor nerve conduction velocity (MNCV) was measured using neuroelectrophysiological techniques. The L 4-6 segments of the spinal cord were collected, and histopathological changes were observed through HE staining. The co-localization of leucine-rich repeat kinase 2 (LRRK2) in the spinal dorsal horn with the microglial activation marker ionized calcium-binding adaptor molecule-1 (Iba-1) was observed by immunofluorescence staining. The L 4-6 segments of the spinal dorsal horn were harvested for determination of the level of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (using enzyme-linked immunosorbent assay) and expression of LRRK2, Iba-1, nuclear factor kappa B (NF-κB) p65 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot). Results:Compared to C group, the TWL was significantly shortened, the MNCV was decreased, the percentage of cells co-expressing LRRK2 and Iba-1 in the spinal dorsal horn was increased, the contents of TNF-α, IL-1β and IL-6 were increased, the expression of LRRK2 and Iba-1 was up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was increased ( P<0.05), and histological examination revealed nuclear shrinkage and inflammatory cell infiltration in the spinal dorsal horn tissue in DNP group. Compared with DNP group, the MWT was significantly increased, the TWL was prolonged, the MNCV was elevated, the percentage of cells co-expressing LRRK2 and Iba-1 was decreased, the contents of TNF-α, IL-1β and IL-6 were decreased, the expression of LRRK2 and Iba-1 was down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was decreased ( P<0.05), and pathological changes of the spinal dorsal horn tissue were significantly attenuated in MLi-2 group. Compared to MLi-2 group, the MWT was significantly decreased, the TWL was shortened, the MNCV was decreased, the percentage of cells co-expressing LRRK2 and Iba-1 was increased, the contents of TNF-α, IL-1β and IL-6 were increased, the expression of Iba-1 was up-regulated, and the ratio of p-NF-κB p65/NF-κB p65 was increased in MLi-2+ PMA group ( P<0.05). Conclusions:LRRK2 in spinal dorsal horn may be involved in the maintenance of DNP by promoting the inflammatory response mediated by microglial activation, and the mechanism may be related to the activation of NF-κB signaling pathway in rats.