Xenon ameliorates chronic post-surgical pain by regulating mitophagy in microglia and rats mediated by PINK1/Parkin pathway.

Background: Chronic post-surgical pain (CPSP) is one of the important causes of poor postoperative outcomes, the activation of microglia in the spinal cord is closely related to the generation, transmission and maintenance of CPSP. Xenon (Xe), an anesthetic gas, has been reported to be able to significantly reduce intraoperative analgesia and postoperative pain sensation at low doses. However, the mechanism of the regulatory effect of xenon on activated microglia after CPSP remains unclear. Methods: In this study, CPSP model rats were treated with 50% Xe inhalation for 1 h following skin/muscle incision and retraction (SMIR), once a day for 5 consecutive days, and then the painbehavioraltests (pain behavior indexes paw withdrawal mechanical threshold, PWMT and thermal withdrawal latency, TWL), microglial activation, oxidative stress-related indexes (malondialdehyde, MDA; superoxide dismutase, SOD; hydrogen peroxide, H2O2; and catalase, CAT), mitophagy and PINK1/Parkin pathway were examined. Results: The present results showed that a single dose of Xe treatment in SMIR rat model could significantly improve PWMT and TWL in the short-term at a single treatment and long-term at multiple treatments. Xe treatment inhibited microglia activation and oxidative stress in the spinal dorsal horn of SMIR rats, as indicated by the decrease of Iba1 and MDA/H2O2 levels and the increase of SOD/CAT levels. Compared with the control group, Xe further increased the CPSP promoted Mito-Tracker (a mitochondrial marker) and LC3 (an autophagy marker) co-localization positive spots and PINK1/Parkin/ATG5/BECN1 (autophagy-related proteins) protein expression levels, and inhibited the Mito-SOX (a mitochondrial reactive oxygen species marker) positive signal, indicating that Xe promoted microglia mitophagy and inhibited oxidative stress in CPSP. Mechanistically, we verified that Xe promoted PINK1/Parkin signaling pathway activation. Conclusion: Xe plays a role in ameliorating chronic post-surgical pain by regulating the PINK1/Parkin pathway mediated microglial mitophagy and provide new ideas and targets for the prevention and treatment of CPSP.

Effect of Thoracic Epidural Anesthesia on Perioperative Neutrophil Extracellular Trapping Markers in Patients Undergoing Anesthesia and Surgery for Colorectal Cancer: A Randomized, Controlled Trial.

BACKGROUND: Neutrophil extracellular trapping (NETosis) is an immunologic mechanism strongly linked with increased metastatic risk in colorectal cancer. The authors hypothesized that patients who received propofol-epidural anesthesia (PEA) would exhibit decreases in the expression of serum neutrophil myeloperoxidase (MPO) and citrullinated histone H3 (H3Cit) levels compared with patients who received general anesthesia (GA). METHODS: Colorectal cancer surgery patients were randomly assigned to the PEA (n = 30) or GA (n = 30) group. Serum MPO, H3Cit, and metalloproteinase-9 (MMP-9) levels before surgery and 24 h after surgery were measured, and visual analogue scale (VAS) scores were recorded. RESULTS: The patients who received PEA showed decreases in MPO (28.06 ± 11.23 vs 20.54 ± 7.29 ng/ ml; P = 0.004) and H3Cit [3.22 ± 0.86 vs 2.73 ± 0.94 ng/ ml; P = 0.042) 24 h after surgery compared with the patients subjected to GA. In addition, there was no difference in MMP-9 levels (75.98 ± 26.9 vs 73.45 ± 28.4 ng/ ml; P = 0.726). The visual analogue scale scores 2 h and 24 h after operation were significantly lower in PEA group (P < 0.05). The number of postoperative analgesia pump pressings and sufentanil consumptions within 48 h after surgery were significantly lower in the PEA group (P < 0.001). CONCLUSIONS: Propofol-epidural anesthesia reduces the expression of NETosis (MPO and H3Cit) in serum during colorectal cancer surgery. CLINICAL TRIAL REGISTRATION: ChiCTR2200066708 ( www.chictr.org.cn ).

Application Effect of Dexmedetomidine and Dezocine in Patients Undergoing Lung Cancer Surgery under General Anesthesia and Analysis of Their Roles in Recovery Time and Cognitive Function.

Objective: To explore the application influence of dexmedetomidine (DEX) and dezocine in patients undergoing lung cancer surgery under general anesthesia and analysis of their roles in recovery time and cognitive function. Methods: A total of 120 patients who accepted thoracoscopic pulmonary wedge resection in our hospital from November 2021 to April 2022 were selected and randomly divided into group A (n =60) and group B (n =60). DEX combined with dezocine-assisted anesthesia was performed to group A, and the equal dose of normal saline was administered to group B, so as to compare their inflammatory influence level, brain function, arterial blood gas index, and cognitive function. Results: Compared with group B, group A obtained significantly lower intraoperative and postoperative inflammatory factor levels (P < 0.001), better postoperative brain function and arterial blood gas index (P < 0.001), and lower Loewenstein Occupational Therapy Cognitive Assessment (LOTCA) scores after surgery (P < 0.001). Combining DEX with dezocine-assisted general anesthesia can improve the inflammatory factors level of patients undergoing lung cancer surgery and maintain their brain function and oxygen saturation, so that they have better postoperative cognitive function. Therefore, such anesthesia modality should be promoted in practice.

The Protective Effects of Ropivacaine Against High Glucose-induced Brain Microvascular Endothelial Injury by Reducing MMPs and Alleviating Oxidative Stress.

Diabetes is undoubtedly affecting global health. Considerable attention has been directed to brain complications caused by diabetes, which are reported to be related to the injury on brain microvascular endothelial cells. Oxidative stress and degradation of vascular basement membrane contribute to the injury of vascular endothelia by diabetes. The present study aims to investigate the effects of ropivacaine on high glucose-induced brain microvascular endothelial injury, as well as the underlying mechanism. Cell viability was determined by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The production of reactive oxygen species (ROS) was evaluated by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. Quantitative real-time PCR (QRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to determine the expression levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), intercellular cell adhesion molecule-1 (ICAM-1), and vascular endothelial growth factor (VEGF). The production of nitric oxide (NO) was detected by DAF-FM DA staining. The expression of inducible nitric oxide synthase (iNOS) was evaluated by qRT-PCR and Western blot analysis. Western blot was used to determine the expression of nuclear factor erythroid 2-related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1).The cell viability of bEnd.3 brain endothelial cells was inhibited by high glucose, which was rescued by ropivacaine. The elevated production of ROS and MDA by high glucose was reversed by ropivacaine. Ropivacaine suppressed the expression of up-regulated iNOS, NO, MMP-2, MMP-9, ICAM-1, and VEGF induced by high glucose incubation. The expression of Nrf-2 and HO-1 by high glucose incubation was significantly inhibited by ropivacaine treatment.Ropivacaine might alleviate high glucose-induced brain microvascular endothelial injury by suppressing oxidative stress and down-regulating MMPs.

Neuroprotective Effects Against Cerebral Ischemic Injury Exerted by Dexmedetomidine via the HDAC5/NPAS4/MDM2/PSD-95 Axis.

Numerous evidences have highlighted the efficient role of dexmedetomidine (DEX) in multi-organ protection. In the present study, the neuroprotective role of DEX on cerebral ischemic injury and the underlining signaling mechanisms were explored. In order to simulate cerebral ischemic injury, we performed middle cerebral artery occlusion in mice and oxygen-glucose deprivation in neurons. Immunohistochemistry, Western blot analysis, and RT-qPCR were used to examine expression of HDAC5, NPAS4, MDM2, and PSD-95 in hippocampus tissues of MCAO mice and OGD-treated neurons. MCAO mice received treatment with DEX and sh-PSD-95, followed by neurological function evaluation, behavioral test, infarct volume detection by TTC staining, and apoptosis by TUNEL staining. Additionally, gain- and loss-of-function approaches were conducted in OGD-treated neuron after DEX treatment. Cell viability and apoptosis were assessed with the application of CCK-8 and flow cytometry. The interaction between MDM2 and PSD-95 was evaluated using Co-IP assay, followed by ubiquitination of PSD-95 detection. As per the results, HDAC5 and MDM2 were abundantly expressed, while NPAS4 and PSD-95 were poorly expressed in hippocampus tissues of MCAO mice and OGD-treated neurons. DEX elevated viability, and reduced LDH leakage rate and apoptosis rate of OGD-treated neurons, which was reversed following the overexpression of HDAC5. Moreover, HDAC5 augmented MDM2 expression via NPAS4 inhibition. MDM2 induced PSD-95 ubiquitination and degradation. In MCAO mice, DEX improved neurological function and behaviors and decreased infarct volume and apoptosis, which was negated as a result of PSD-95 silencing. DEX plays a neuroprotective role against cerebral ischemic injury by disrupting MDM2-induced PSD-95 ubiquitination and degradation via HDAC5 and NPAS4.

Calpain-2 plays a pivotal role in the inhibitory effects of propofol against TNF-α-induced autophagy in mouse hippocampal neurons.

Calpains are calcium-dependent proteases and play critical roles in neuronal autophagy induced by inflammation. Propofol has been reported to exert anti-inflammatory effects in neurons. We aimed to identify whether and how propofol-modulated calpain activity and neuron autophagy in response to tumour necrosis factor-α (TNF-α). Mouse hippocampal neurons were pre-treated with propofol and exposed to TNF-α. Autophagy was evaluated by fluorescent autophagy assay and by measuring LC3I and LC3II expression. Intracellular calcium concentration was measured by fluorescent assay. Calpain activation was measured by calpain activity assay. The protein expression of intracellular signalling molecules was detected by Western blot analysis. Compared with untreated control neurons, 40 ng/mL TNF-α treatment for 2 hours induced neuron autophagy, which was attenuated by 25 µmol/L propofol. TNF-α induced intracellular calcium accumulation, the phosphorylation of calcium/calmodulin-dependent protein kinase II (CAMK II) and calpain-2, calpain activation and lysosomal cathepsin B release as well as tyrosine kinase receptor B (TrkB) truncation. These effects were alleviated by propofol, calcium chelator, CAMK II inhibitor, calpain-2 inhibitor, calpain-2 siRNA transfection and N-Methyl-d-aspartic acid (NMDA) receptor antagonist. Propofol, via NMDA receptor, inhibited TNF-α-mediated hippocampal neuron autophagy. The mechanism may involve calcium and calcium-dependent signalling pathway, especially CAMK II and calpain-2.

Preoperative leukocytosis is associated with increased tumor-infiltrating neutrophil extracellular traps and worse outcomes in esophageal cancer.

BACKGROUND: We evaluated the correlation between preoperative white blood cell (WBC) count and the prognosis in esophageal cancer (EC) patients who underwent esophagectomy, and explored the potential link between preoperative WBC count and tumor-infiltrating neutrophil extracellular traps (NETs) in EC. METHODS: From January 2013 to December 2017, 3,096 patients at Fudan University Shanghai Cancer Center (FUSCC) undergoing esophagectomy for EC were enrolled in this retrospective cohort. The prognostic value of preoperative WBC count together with tumor-infiltrating NETs was investigated. RESULTS: Leukocytosis (≥10,000/µL) was significantly associated with decreased overall survival (OS) and disease-free survival (DFS) (P<0.05). Further, moderate leukocytosis (≥7,000/µL) were also identified as an independent prognostic factor for survival. Additionally, moderate leukocytosis was correlated with male sex (P=0.006), advanced T stage (P<0.001), TNM stage (P<0.001) and ineffective postoperative chemotherapy (P<0.001), and moderate leukocytosis even predicted increased relapse postoperatively (P<0.001). Importantly, patients with moderate leukocytosis had a significantly higher level of intra-tumoral NETs infiltration (P<0.001), and the higher level of NETs infiltration were associated with worse OS and DFS (P<0.001). CONCLUSIONS: Our data indicated that preoperative moderate leukocytosis is associated with increased tumor-infiltrating NETs and is an independent prognostic factor for survival in EC after surgery.

Analgesic Effect of Methane Rich Saline in a Rat Model of Chronic Inflammatory Pain.

How oxidative stress contributes to neuro-inflammation and chronic pain is documented, and methane is reported to protect against ischemia-reperfusion injury in the nervous system via anti-inflammatory and antioxidant properties. We studied whether methane in the form of methane rich saline (MS) has analgesic effects in a monoarthritis (MA) rat model of chronic inflammatory pain. Single and repeated injections of MS (i.p.) reduced MA-induced mechanical allodynia and multiple methane treatments blocked activation of glial cells, decreased IL-1ß and TNF-α production and MMP-2 activity, and upregulated IL-10 expression in the spinal cord on day 10 post-MA. Furthermore, MS reduced infiltrating T cells and expression of IFN-γ and suppressed MA-induced oxidative stress (MDA and 8-OHDG), and increased superoxide dismutase and catalase activity. Thus, MS may offer anti-inflammatory and antioxidant effects to reduce chronic inflammatory pain.

Protective effects of oridonin on the sepsis in mice.

This study aimed to investigate the protective effects of oridonin (ORI) on cecal ligation and puncture (CLP)-induced sepsis in mice. Male C57BL/6 mice weighing 22-30 g and aged 8-10 weeks were randomly assigned to three groups: Sham group, CLP group, or CLP plus ORI group. In the CLP group and ORI group, CLP was induced, and intraperitoneal injection of normal saline and oridonin (100 µg/kg) was conducted, respectively. The survival rate was determined within the following 7 days. The blood, liver, and lung were collected at 24 hours after injury. Hematoxylin-eosin staining of the lung, detection of lung wet-to-dry ratio, and serum cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6), and examination of intraperitoneal and blood bacterial clearance were conducted to evaluate the therapeutic efficacy. Results showed that ORI treatment significantly reduced the lung wet-to-dry ratio, decreased serum TNF-α and IL-6, and improved liver pathology compared with the CLP group (p < 0.05). Moreover, the intraperitoneal and blood bacterial clearance increased markedly after ORI treatment (p < 0.05). The 7-day survival rate in the ORI group was also dramatically higher than in the CLP group (p < 0.05). Our findings indicate that ORI can attenuate liver and lung injuries and elevate bacterial clearance to increase the survival rate of sepsis mice.

Application of hyperbaric oxygen in liver transplantation.

In recent years, hyperbaric oxygen (HBO) has been used in the treatment of a lot of diseases such as decompression sickness, arterial gas embolism, carbon dioxide poisoning, soft tissue infection, refractory osteomyelitis, and problematic wound, but little is known about its application in liver transplantation. Although several studies have been conducted to investigate the protective effects of HBO on liver transplantation and liver preservation, there are still some controversies on this issue, especially its immunomodulatory effect. In this short review, we briefly summarize the findings supporting the application of HBO during liver transplantation (including donors and recipients).

Preparation, physicochemical characteristics and bioavailability studies of an atorvastatin hydroxypropyl-beta-cyclodextrin complex.

The aim of this study was to improve the solubility, stability and bioavailability of amorphous atorvastatin calcium (AT) by complexing it with hydroxypropyl-beta-cyclodextrin. The formation of the inclusion complexation was identified by molecular modeling, phase solubility diagrams, differential scanning calorimetry and X-ray powder diffractometry. Orally Disintegrating Tablets (ODT) were then manufactured by direct compression. Apart from improved stability compared to pure AT, disintegration time of 27s, hardness of 5 kg and favorable mouth feel were achieved. In vitro dissolution tests of the ODT of AT inclusion complex exhibited higher dissolution rates than those with pure drug and the commercial tablet Lipitor. In vivo bioavailability studies in rats also showed shorter T(max), higher C(max) and increased AUC of 4.42 and 1.86 fold compared to the plain drug ODT and Lipitor. These results strongly suggest to use HP-beta-CD to improve the physicochemical characteristics and bioavailability of AT.