The pathogenesis of acute lung injury (ALI), sepsis, and ulcerative colitis (UC) is closely interrelated. As inflammatory diseases, they are all characterized by the excessive production of inflammatory cytokines. Therefore, suppression of these cytokines represents a promising therapeutic strategy. Herein, we designed and synthesized a series of N-(4-(pyridin-4-yloxy)phenyl)-1,4-dihydropyridazine-3-carboxamide derivatives based on integrating three structural fragments relating to anti-inflammation. The anti-inflammatory activities of these compounds were evaluated in J774A.1 macrophages, and most compounds effectively inhibited interleukin-6 (IL-6) release in vitro. Compound z18 was identified as the most promising candidate and was further investigated for its dose-dependent effects. Preliminary mechanistic studies indicated that compound z18 suppresses c-Kit kinase activity and subsequently inhibits the NF-κB signaling pathway. z18 exhibited significant anti-inflammatory effects in vivo in LPS-induced ALI, sepsis, and UC mouse models. The preliminary drug-like properties of z18 were also evaluated. z18 demonstrated favorable stability in body fluid stability studies, and its favorable safety profile was further confirmed by subacute toxicity studies. In summary, this study offers novel insights into the design of anti-inflammatory agents and presents z18 as a promising candidate for the treatment of ALI, sepsis, and UC.

This study aimed to determine whether ultrasound-guided pneumatic tourniquet pressure setting reduces pulmonary ventilation dysfunction and enhances hemodynamic stability in patients undergoing lower-limb surgery. This was a prospective, single-center, assessor-blinded, randomized controlled trial. Forty-four patients scheduled for lower-limb surgery requiring tourniquet application were assessed for eligibility. Two patients were excluded because of protocol deviations (one received combined sedative medication and one was converted to general anesthesia intraoperatively). The remaining 42 patients were randomly allocated to a control group and an ultrasound-guided group (n = 21 each). In the ultrasound-guided group, the optimal tourniquet pressure was defined as the pressure at which the popliteal arterial blood flow ceased under ultrasonographic imaging. The control group underwent the conventional pressure-setting method. The primary outcome was the arterial-to-alveolar oxygen partial pressure ratio (A/a ratio) at 6 h after tourniquet deflation (T4). Secondary outcomes included mean arterial pressure (MAP), heart rate (HR), plasma malondialdehyde (MDA) and superoxide dismutase (SOD) concentrations, arterial oxygen partial pressure (PaO₂), arterial carbon dioxide partial pressure (PaCO₂), respiratory index (RI), alveolar-arterial oxygen partial pressure difference (PA–aDO₂). All outcomes, including A/a ratio, were measured at designated time points: immediately before tourniquet inflation (T0), and at 1 h (T1), 0.5 h (T2), 2 h (T3), 6 h (T4), and 24 h (T5) after inflation. Compared with the control group, the ultrasound-guided group exhibited higher MAP and lower HR at T2. At T4, PaO₂ and A/a ratio increased, while PA–aDO₂ and RI decreased. From T3 to T5, MDA concentration was significantly lower and SOD concentration significantly higher in the ultrasound-guided group compared with the control group. Ultrasound-guided tourniquet pressure setting enhances hemodynamic stability and reduces pulmonary ventilation dysfunction in patients undergoing lower-limb surgery. These findings were accompanied by lower MDA levels and higher SOD activity, suggesting a potential association with reduced oxidative stress during ischemia–reperfusion. The study was registered in the Chinese Clinical Trial Register website (www.chictr.org.cn, ChiCTR1900026552,2019/10/13).

Myeloid differentiation factor 88 (MyD88) is a critical mediator of inflammatory signaling. However, the development of MyD88 inhibitors has been limited, and there are no reports on MyD88 degraders. Based on our previously identified MyD88 inhibitor c17 and employing a template-assisted modification strategy, we designed and synthesized 40 novel derivatives, and identified the optimal compound d21, which could inhibit nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway activation by facilitating the interaction between ring finger protein 126 (RNF126) (an E3 ubiquitin ligase) and MyD88, thereby promoting ubiquitination and subsequent degradation of MyD88. The subacute toxicity test indicated that d21 had good safety profile. Furthermore, it demonstrated significant anti-inflammatory effects in both cecal ligation and puncture (CLP) and lipopolysaccharide (LPS) induced acute lung injury (ALI) models, effectively ameliorating ALI symptoms. These findings suggest that d21 holds considerable promise as a MyD88-targeting molecular glue degrader for the treatment of ALI.

Background: Cerebral ischemia-reperfusion injury (CIRI) is a severe neurological condition where restoring neuronal mitochondrial function critically impacts prognosis. While electroacupuncture (EA) has demonstrated neuroprotective effects by improving mitochondrial function, the precise underlying mechanisms remain unclear. Emerging evidence suggests that astrocyte-to-neuron mitochondrial transfer, facilitated by mitochondrial Rho-GTPase 1 (Miro1), serves as a vital neuroprotective pathway. Therefore, this study investigates whether astrocytic Miro1 participates in the neuroprotective effects of EA against CIRI in mice by regulating the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 40 (TOM40) and adenosine triphosphate (ATP) levels in damaged neurons. Methods: 126 C57BL/6 mice were randomly allocated into seven experimental groups (n = 18 per group): Sham-operated (Sham), middle cerebral artery occlusion (MCAO) model, EA, sham electroacupuncture (SEA), EA combined with astrocyte-specific Miro1 knockdown (GFAP: glial fibrillary acidic protein, EA+AAV-GFAP-shMiro1), astrocyte-specific Miro1 over-expression (AAV-GFAP-hiMiro1), and adenoviral empty vector control (AAV-GFAP-control). The CIRI model was induced using MCAO. Prior to model induction, the EA group received pretreatment with EA at the Baihui (GV20) acupoint. The SEA group underwent identical procedures to the EA group except for electrical stimulation. For the EA+AAV-GFAP-shMiro1, AAV-GFAP-hiMiro1, and AAV-GFAP-control groups, mice received intracerebroventricular injections of AAV-GFAP-shMiro1, AAV-GFAP-hiMiro1, or AAV-GFAP-control, respectively, 48 hours prior to EA treatment, with other procedures matching the EA group. At 24 hours post-reperfusion, neurological deficit scores, cerebral infarct volume, and neuronal survival in the peri-infarct penumbra were assessed. Astrocytes and neurons from the peri-infarct penumbra were isolated to measure ATP levels and expression of the mitochondrial-specific protein TOM40 in neurons, as well as ATP levels, TOM40, and Miro1 protein expression in astrocytes. Results: Relative to the Sham group, the MCAO group displayed a significant increase in cerebral infarct volume and neurological deficit scores, accompanied by a marked reduction in neuronal viability, TOM40 expression, and ATP levels (p < 0.01). In contrast to the MCAO and SEA groups, the EA and AAV-GFAP-hiMiro1 groups demonstrated improved neurological scores, reduced infarct volume, enhanced neuronal viability, elevated neuronal ATP levels and TOM40 expression, as well as decreased astrocytic ATP and TOM40 levels, but significantly increased Miro1 expression in astrocytes (p < 0.01). When compared to the EA group, the EA+AAV-GFAP-shMiro1 group exhibited a reversal of all the aforementioned improvements (p < 0.01), while the AAV-GFAP-hiMiro1 group showed no significant changes (p > 0.05). Conclusions: EA exerts neuroprotective effects in MCAO mice by upregulating Miro1 protein expression in astrocytes and upregulating the mitochondrial marker TOM40 alongside ATP levels in neurons. Silencing Miro1 abolished the neuroprotective effects of EA and reduced neuronal TOM40 expression, while Miro1 overexpression increased this mitochondrial marker and mimicked EA-mediated neuroprotection. These findings identify Miro1 as a key effector of EA-induced neuroprotection, although the upstream signaling pathways linking EA to Miro1 upregulation require further investigation.

Purpose:Respiratory management remains a major challenge during painless endoscopy in overweight or obese patients because of altered body fat distribution and reduced pulmonary reserve. This study evaluated whether ciprofol could reduce respiratory adverse events compared with propofol during painless gastroscopy in this population. Methods:In this single-center, prospective, single-blind randomized controlled trial, 106 overweight or obese patients undergoing painless gastroscopy were randomly assigned (1:1) to receive either propofol (2 mg/kg) or ciprofol (0.4 mg/kg) following intravenous alfentanil (5 μg/kg). The primary outcome was the overall incidence of respiratory adverse events from anesthesia induction to completion of endoscopy, including hypoxemia (SpO2 <90% for >15 s), respiratory depression (respiratory rate <8 breaths/min for >30 s), and apnea (absence of chest wall movement for >20 s). Results:A total of 106 patients completed the study (53 per group). The incidence of respiratory adverse events was significantly lower in the ciprofol group than in the propofol group (33.96% vs. 66.04%, P=0.001). The lowest intraoperative SpO2 was significantly higher in the ciprofol group (93.50% vs. 89.00%, P=0.044). Compared with ciprofol, propofol was associated with higher incidences of hypotension (22.64% vs. 7.55%, P=0.030) and injection pain (18.87% vs. 0%, P=0.001). No significant differences were observed in other adverse events, including nausea and vomiting. Conclusion:In overweight or obese patients undergoing painless gastroscopy, ciprofol at 0.4 mg/kg significantly reduced respiratory adverse events, improved oxygenation, and decreased injection pain compared with propofol. These findings suggest that ciprofol may be a preferable anesthetic option for painless endoscopy in this population. Clinical Trial Registration:ChiCTR2600117367.

Peroxiredoxin 1 (PRDX1) is a member of the peroxidase family of antioxidant enzymes. However, the role and mechanism of PRDX1 in acrylamide (ACR)-induced nerve damage have not been reported. We used SD rats and well-differentiated rat pheochromocytoma cells (PC-12 cells) to established in vivo and in vitro models of ACR. Immunohistochemistry, immunofluorescence and RT-qPCR experiments were used to detect the expression of PRDX1 in neurons of rat hippocampal tissue. The ultrastructural changes of neurons and PC-12 cells in rat hippocampal tissue were observed under transmission electron microscope. Western blot detected the protein expression levels of PRDX1, PTEN, AKT and p-AKT. In vivo and in vitro experimental results showed that PRDX1 showed a significant up-regulation trend after ACR exposure (p < 0.05). In vitro experiments showed that after inhibiting PRDX1 expression with PRDX1 siRNA, the survival rate of PC-12 cells significantly increased, and the damage to cell morphology and organelles was markedly improved. Western blot analysis revealed that ACR exposure can cause a significant increase in PTEN protein expression level and p-AKT/AKT protein ratio (p < 0.05). After inhibiting the expression of PRDX1, the protein expression level of PTEN and the protein ratio of p-AKT/AKT were significantly reduced, while the protein levels of SYN1 and BDNF were significantly increased (p < 0.05). This study, for the first time, demonstrates that PRDX1 affects ACR-induced neurotoxicity by regulating the PTEN/AKT signaling pathway. And, provides novel insights into the prevention and treatment of neurotoxicity in populations exposed to ACR.

Purpose:Laparoscopic hysterectomy(LH) is often associated with multimodal postoperative pain, which impedes patient recovery. This study aimed to evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on postoperative acute visceral,incisional, and low back pain (LBP) and recovery in patients undergoing LH. Patients and Methods:Patients scheduled for elective LH at the First Affiliated Hospital of Wenzhou Medical University (February 2024-2025) were randomly divided (1:1 ratio) into a TEAS and control group. TEAS involved bilateral stimulation at Hegu-Neiguan, and Sanyinjiao-Zusanli 30 min before anesthesia induction and throughout surgery, while the control involved electrodes placed identically for sham stimulation. Pain intensity (visceral, incisional, and LBP) was evaluated using numerical rating scale on postoperative days (PODs) 0 (day of surgery), 1, and 2. Secondary outcomes comprised postoperative serum cytokine profiles, opioid consumption, rescue analgesia demands on POD 1, adverse events, and standardized recovery metrics. Results:The TEAS group (n=45) demonstrated superior pain control compared to the control group (n=48), with significantly lower visceral pain scores (POD 0-1), decreased LBP scores (POD 0-2), and reduced incidence of moderate-to-severe visceral pain (POD 0-2) and LBP (POD 0) (all P < 0.017). TEAS resulted in lower interleukin-6 levels, total sufentanil consumption, and rescue analgesia demands on POD 1(all P < 0.05). TEAS was associated with a shorter time to pelvic drain removal, decreased postoperative hospitalization, earlier ambulation, and lower incidence of postoperative nausea and vomiting (all P < 0.05). No significant improvement in incisional pain was observed with TEAS intervention. Conclusion:TEAS provided differential postoperative analgesia, effectively alleviating visceral and LBP but not incisional pain. This primary benefit, coupled with reduced inflammation, opioid use, and adverse events, facilitated recovery in LH patients. These findings support the incorporation of TEAS as an effective non-pharmacological adjuvant within multimodal analgesia and ERAS protocols.

Background: Ischemic stroke is a serious clinical condition that is challenging to cure; therefore, slowing down the depletion of ATP is crucial to enhancing the tolerance of ischemic tissue through preconditioning. Electroacupuncture (EA) preconditioning induces tolerance to cerebral ischemia; however, the underlying mechanism remains unclear. Objective: The P2×7 receptor (P2×7R) mediates the stimulation of microglial cells and is involved in the development of cerebral ischemia-reperfusion (I/R) damage. We hypothesized that the protective effect of EA preconditioning is associated with the downregulation of P2×7R expression. Methods: We performed EA at the ''Baihui'' and ''Fengfu'' for 30 min before establishing a rat model of cerebral I/R induced based on the middle cerebral artery occlusion model (MCAO). MCAO rats were administered a ventricular injection of 2 '(3′)-O-(4-benzoyl) adenosine triphosphate (BzATP), a P2×7R agonist, 30 min before EA. Neurologic scoring, infarction volume, and expression of cytokines, Bcl-2 and Bax, Iba1, P2×7R, p38, and phosphorylated p38 (p-p38) in ischemia penumbra were detected 24 h after cerebral I/R. Results: EA preconditioning ameliorated neurologic scoring, decreased infarction volume, and neuronal injury, and decreased cytokine release, while BzATP exacerbated cerebral I/R damage and inflammation events, unlike the favorable efficacy of EA. EA inhibited the expression of Iba-1, P2×7R, and p-p38/p38 in the ischemic penumbra, whereas BzATP reversed this effect. Conclusions: EA could induce cerebral tolerance to I/R damage by suppressing P2×7R expression and release of inflammatory factors.

Cerebral ischemia-reperfusion injury (CIRI) involves innate immunity activation in astrocytes and the inflammatory response. The interferon-induced transmembrane protein 3 (IFITM3) is an immune protein whose role in CIRI remains largely unexplored. This study investigated the role of IFITM3 in CIRI in mice. Adeno-associated virus (AAV)-mediated delivery of siRNA was used to inhibit IFITM3 expression, assessing effects on astrocyte activation, inflammatory cytokine expression. Primary cultured astrocytes were exposed to oxygen-glucose deprivation/reperfusion (OGD/R) to study IFITM3's role in vitro. Western blotting (WB) was employed to measure C-reactive protein (CRP) and IFITM3 levels, and enzyme-linked immunosorbent assay (ELISA) was used to quantify inflammatory cytokines. γ-Secretase activity, as well as Aβ40 and Aβ42 peptide levels, were measured to evaluate its activity. IFITM3 expression in astrocytes was significantly elevated following CIRI (p < 0.001), leading to increased γ-secretase activity and higher production of Aβ40 and Aβ42 peptides (p < 0.001). CRP levels were also upregulated in the context of IFITM3 expression (p < 0.01). Inhibiting IFITM3 expression via AAV-mediated delivery of siRNA significantly reduced astrocyte activation, inflammatory cytokine expression (p < 0.001 for IL-1β, IL-6, TNF-α, and IFN-γ), and improved neurobehavioral scores (p < 0.001). In vitro, IFITM3 inhibition significantly reduced the protein and mRNA levels of IFITM3, suppressed astrocyte activation, and decreased the expressions of inflammatory factors (p < 0.01). IFITM3 inhibition reduced the apoptosis of co-cultured neuronal cells (p < 0.01) and suppressed TLR4/NF-κB expression (p < 0.01), thereby attenuating the production of inflammatory factors. Inhibiting IFITM3 expression in astrocytes not only regulates γ-secretase activity but also mitigates neuroinflammation, thereby alleviating CIRI.

Objective:The aim of this study is to evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) during the perioperative period on the quality of postoperative recovery among patients undergoing pterygium excision. Methods:A total of 110 patients scheduled for unilateral pterygium excision were enrolled and randomly assigned in equal numbers to the TEAS group or the control group. In the TEAS group, patients received TEAS at the LI4 and PC6 acupoints, initiated 30 min before anesthesia induction and continued until the conclusion of surgery. In the control group, patients had electrode pads applied without active stimulation. Numerical Rating Scale (NRS) scores, State-Trait Anxiety Inventory (S-TAI) scores, and Quality of Recovery-40 Questionnaire (QoR-40) scores were collected from both groups. Results:No statistically significant differences were observed in baseline demographic and clinical characteristics between the two groups. At 24 h postoperatively, patients in the TEAS group demonstrated significantly higher QoR-40 scores and significantly lower NRS pain scores and postoperative SAI scores compared to the control group. Conclusion:TEAS was effective in reducing postoperative pain and anxiety levels while enhancing the quality of postoperative recovery in patients undergoing pterygium surgery. Clinical trial registration:https://www.chictr.org.cn, identifier ChiCTR2200056062.

Background This study was identified the risk factors for and designed to investigate influence of postoperative moderate-to-severe pain of post anaesthesia care unit (PACU) in patients with malignancy. Methods A retrospective study was performed on 22,600 cancer patients with malignancy who underwent elective radical surgery in the new hospital of First Affiliated Hospital of Wenzhou Medical University, between January 2016 and June 2021. All patients were transferred to the PACU after tracheal extubation. Patients were divided into two groups according to a visual analogue scale (VAS) score of > 3: the no-moderate-severe-pain group and moderate-to-severe-pain group. Data pertaining to demographic, surgical, anaesthetic, and other factors were recorded. Lasso and logistic regression analysis was performed to explore the risk factors, then a nomogram was constructed to predict the moderate-severe-pain in the PACU. Validation was performed by using another 662 cancer patients in old hospital. The ROC curves and calibration curve were used to evaluate the accuracy and predictive ability of the nomogram. Results The incidence of postoperative moderate-to-severe pain of PACU in patients with malignancy was 1.42%. Gender, type of surgery, postoperative use of PCA, intraoperative adjuvant opioid agonists, NSAIDS, epidural analgesia, duration of anaesthesia, intraoperative massive haemorrhage, PACU vomiting were independent predictors for postoperative moderate-to-severe pain of PACU in the patients with malignancy. The area under the ROC curve of the predictive models in the primary and validation groups were 0.817 and 0.786, respectively. Moderate-to-severe pain in the PACU correlated with hypertension, hyperglycaemia, agitation, and hypoxemia ( P < 0.05). Conclusions The prediction model for postoperative moderate-to-severe pain of PACU in patients with malignancy has good predictive ability and high accuracy, which is helpful for PACU medical staff to identify and prevent postoperative moderate-to-severe pain in advance. Trial registration The study was approved by the Clinical Research Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University (No.KY2021-097) and registered in the Chictr.org.cn registration system on 06/12/2021 (ChiCTR2100054013).

BACKGROUND Acupuncture promotes the recovery of gastrointestinal function and provides analgesia after major abdominal surgery. The effects of transcutaneous electrical acupoint stimulation (TEAS) remain unclear. AIM To explore the potential effects of TEAS on the recovery of gastrointestinal function after gastrectomy and colorectal resection. METHODS Patients scheduled for gastrectomy or colorectal resection were randomized at a 2:3:3:2 ratio to receive: (1) TEAS at maximum tolerable current for 30 min immediately prior to anesthesia induction and for the entire duration of surgery, plus two 30-min daily sessions for 3 consecutive days after surgery (perioperative TEAS group); (2) Preoperative and intraoperative TEAS only; (3) Preoperative and postoperative TEAS only; or (4) Sham stimulation. The primary outcome was the time from the end of surgery to the first bowel sound. RESULTS In total, 441 patients were randomized; 405 patients (58.4 +/- 10.2 years of age; 247 males) received the planned surgery. The time to the first bowel sounds did not differ among the four groups (P = 0.90; log-rank test). On postoperative day 1, the rest pain scores differed significantly among the four groups (P = 0.04; Kruskal-Wallis test). Post hoc comparison using the Bonferroni test showed lower pain scores in the perioperative TEAS group (1.4 +/- 1.2) than in the sham sti-mulation group (1.7 +/- 1.1; P = 0.04). Surgical complications did not differ among the four groups. CONCLUSION TEAS provided analgesic effects in adult patients undergoing major abdominal surgery, and it can be added to clinical practice as a means of accelerating postoperative rehabilitation of these patients.

Previous studies have demonstrated that cardiomyocyte apoptosis, ferroptosis, and inflammation participate in the progress of sepsis-induced cardiomyopathy (SIC). Although Islet cell autoantigen 69 (ICA69) is an imperative molecule that could regulate inflammation and immune response in numerous illnesses, its function in cardiovascular disease, particularly in SIC, is still elusive. We confirmed that LPS significantly enhanced the expression of ICA69 in wild-type (WT) mice, macrophages, and cardiomyocytes. The knockout of ICA69 in lipopolysaccharide(LPS)-induced mice markedly elevated survival ratio and heart function, while inhibiting cardiac muscle and serum inflammatory cytokines, reactive oxygen (ROS), and ferroptosis biomarkers. Mechanistically, increased expression of ICA69 triggered the production of STING, which further resulted in the production of intracellular lipid peroxidation, eventually triggering ferroptosis and heart injury. Intriguingly, ICA69 deficiency only reversed the ferroptotic marker levels, such as prostaglandin endoperoxide synthase 2 (PTGS2), malonaldehyde (MDA), 4-hydroxynonenal (4HNE), glutathione peroxidase 4 (GPX4), superoxide dismutase (SOD), iron and lipid ROS, but had no effects on the xCT-dependent manner. Additionally, greater ICA69 level was identified in septic patients peripheralblood mononuclear cells (PBMCs) than in normal control groups. Generally, we unveil that ICA69 deficiency can relieve inflammation and ferroptosis in LPS-induced murine hearts and macrophages, making targeting ICA69 in heart a potentially promising treatment method for SIC.

Previous studies have suggested that miR-324-3p is related to the pathophysiology of cerebral ischemia, but the mechanism underlying this relationship is unclear. In this study, we found that miR-324-3p expression was decreased in patients with acute ischemic stroke and in in vitro and in vivo models of ischemic stroke. miR-324-3p agomir potentiated ischemic brain damage in rats subjected to middle cerebral artery occlusion, as indicated by increased infarct volumes and cell apoptosis rates and greater neurological deficits. In a PC12 cell oxygen-glucose deprivation/reoxygenation model, a miR-324-3p mimic decreased cell viability and expression of the anti-apoptotic protein BCL2 and increased expression of the pro-apoptotic protein BAX and rates of cell apoptosis, whereas treatment with a miR-324-3p inhibitor had the opposite effects. Silencing miR-324-3p increased adenosine A1 receptor (A1R) expression through regulation of GATA binding protein 2 (GATA2). These findings suggest that silencing miR-324-3p reduces ischemic brain damage via the GATA2/A1R axis.

ABSTRACT Background: Acupuncture is a treatment for neuropathic pain, but its mechanism remains unclear. Previous studies showed that analgesia was induced in rats with neuropathic pain when their spinal cord adenosine content increased after electroacupuncture (EA); however, the mechanism behind this electroacupuncture-induced increase has not been clarified. Objective: This study aimed to determine the role that ecto-5’-nucleotidase plays in EA-induced analgesia for neuropathic pain. Methods: We performed electroacupuncture at the Zusanli acupoint on the seventh day after establishing a rat model of neuropathic pain induced through chronic constriction injuries. We observed the mechanical withdrawal threshold and thermal pain threshold and detected the expression of ecto-5’-nucleotidase in the spinal cord using Western blot. Chronic constriction injury rat models were intraperitoneally injected with α,β-methyleneadenosine 5'-diphosphate, an ecto-5’-nucleotidase inhibitor, 30 min before electroacupuncture. The adenosine content of the spinal cord was detected using high-performance liquid chromatography. Lastly, the adenosine A1 receptor agonist N6-cyclopentyladenosine was intrathecally injected into the lumbar swelling of the rats, and the mechanical withdrawal and thermal pain thresholds were reevaluated. Results: Analgesia and increased ecto-5’-nucleotidase expression and adenosine content in the spinal cord were observed 1 h after electroacupuncture. α,β-methyleneadenosine 5'-diphosphate was able to inhibit upregulation of adenosine content and electroacupuncture-induced analgesia. After administration of N6-cyclopentyladenosine, electroacupuncture-induced analgesia was restored. Conclusions: Our results suggest that electroacupuncture at Zusanli can produce analgesia in chronic constriction injury rat models, possibly via the increased ecto-5’-nucleotidase expression induced through electroacupuncture, thus leading to increased adenosine expression in the spinal cord.

Electroacupuncture (EA) is commonly used to treat cerebrovascular diseases. This study aimed to clarify the mechanisms of action of treatments of cerebral ischemic stroke from the perspective of gut microecology. We used a mouse model and cell cultures to investigate the effects of EA on the intestinal microflora in mice models of middle cerebral artery occlusion (MCAO) and the mechanisms underlying the antioxidant activities of metabolites. Fecal microbiota transplantation (FMT) was used to validate the roles of gut microbiota. Metabolomic analysis was performed to characterize the metabolic profile differences between the mice in the EA + MCAO and MCAO groups. Gavaging with feces relieved brain damage in mice that received EA (EA mice) more than in mice that did not (non-EA [NEA] mice). The gut microbial composition and metabolic profiles of the EA and NEA mice were different. In particular, the microbiota from the mice in the EA or EA-FMT groups generated more indole-3-propionic acid (IPA) than the microbiota from the mice in the MCAO or NEA-FMT groups. We confirmed that IPA binds to specific melatonin receptors (MTRs) in target cells and exerts antioxidant effects by adding MTR inhibitors or knocking out the MTR1 gene in vivo and in the oxygen and glucose deprivation/reperfusion models of N2a cell experiments. EA can prevent ischemic stroke by improving the composition of intestinal microbiota in MCAO mice. Moreover, this study reveals a new mechanism of intestinal flora regulation of stroke that differs from inflammation/immunity, namely gut microbiota regulates stroke by affecting IPA levels.

Sepsis-associated encephalopathy (SAE) is a common complication of sepsis caused by neuroinflammation. Electroacupuncture (EA) can be used to treat SAE, but the underlying mechanism is not clear. Lack of PICK1 further aggravates the inflammatory response in mice with sepsis. Therefore, we sought to investigate whether PICK1 is involved in the protective effects of electroacupuncture to SAE. In this study, mice were treated with EA after lipopolysaccharide (LPS) treatment. Behavioral tests; microglial activity of hippocampus; neuron survival and the inflammatory factors PICK1 and TLR4, as well as TLR4-related proteins, such as ERK, JNK, and P38, were assessed after EA treatment. PICK1, TLR4, and TLR4-related proteins, as well as PICK1-TLR4 complex levels were assessed in BV2 cells treated with LPS, PICK1 siRNA, or PICK1 polypeptide. The results indicated that EA could improve neurological assessment and reduce activation of microglial and TLR4 and expression of proinflammatory cytokines. EA also reduced the expression of TLR4 and phosphorylation of ERK/JNK/P38 while, increased the expression of PICK1 and TLR4 complexes. PICK1 knockdown further promoted the expression of TLR4 and phosphorylation of ERK/JNK/P38 in BV2 cells, but this effect was reversed by PICK1 polypeptides. These results suggest that EA may reduce neuroinflammation responses, decrease inflammatory factors, and finally, protect SAE by increasing the formation of PICK1-TLR4 complexes in microglia.

Cerebral ischemic stroke (IS) is still a difficult problem to be solved; energy metabolism failure is one of the main factors causing mitochondrion dysfunction and oxidation stress damage within the pathogenesis of cerebral ischemia, which produces considerable reactive oxygen species (ROS) and opens the blood-brain barrier. Dichloroacetic acid (DCA) can inhibit pyruvate dehydrogenase kinase (PDK). Moreover, DCA has been indicated with the capability of increasing mitochondrial pyruvate uptake and promoting oxidation of glucose in the course of glycolysis, thereby improving the activity of pyruvate dehydrogenase (PDH). As a result, pyruvate flow is promoted into the tricarboxylic acid cycle to expedite ATP production. DCA has a protective effect on IS and brain ischemia/reperfusion (I/R) injury, but the specific mechanism remains unclear. This study adopted a transient middle cerebral artery occlusion (MCAO) mouse model for simulating IS and I/R injury in mice. We investigated the mechanism by which DCA regulates glycolysis and protects the oxidative damage induced by I/R injury through the PDK2-PDH-Nrf2 axis. As indicated from the results of this study, DCA may improve glycolysis, reduce oxidative stress and neuronal death, damage the blood-brain barrier, and promote the recovery of oxidative metabolism through inhibiting PDK2 and activating PDH. Additionally, DCA noticeably elevated the neurological score and reduced the infarct volume, brain water content, and necrotic neurons. Moreover, as suggested from the results, DCA elevated the content of Nrf2 as well as HO-1, i.e., the downstream antioxidant proteins pertaining to Nrf2, while decreasing the damage of BBB and the degradation of tight junction proteins. To simulate the condition of hypoxia and ischemia in vitro, HBMEC cells received exposure to transient oxygen and glucose deprivation (OGD). The DCA treatment is capable of reducing the oxidative stress and blood-brain barrier of HBMEC cells after in vitro hypoxia and reperfusion (H/R). Furthermore, this study evidenced that HBMEC cells could exhibit higher susceptibility to H/R-induced oxidative stress after ML385 application, the specific inhibitor of Nrf2. Besides, the protection mediated by DCA disappeared after ML385 application. To sum up, as revealed from the mentioned results, DCA could exert the neuroprotective effect on oxidative stress and blood-brain barrier after brain I/R injury via PDK2-PDH-Nrf2 pathway activation. Accordingly, the PDK2-PDH-Nrf2 pathway may play a key role and provide a new pharmacology target in cerebral IS and I/R protection by DCA.

Stroke is one of the most important diseases that seriously threaten the health and public health of elderly patients.NSUN2 refers to the predominant methyltransferase for RNA m5C methylation, contributing to increased RNA stability, translocation and translation, and playing an important role in the physiopathology. However, there is insignificant progress on the biological functions and mechanisms of NSUN2 in cerebral ischemia-reperfusion injury. Here, C57BL/6 mice were employed to establish a middle cerebral artery ischemia-reperfusion injury model (MCAO) and found to significantly increase in NSUN2 protein and mRNA expression levels by Western blotting and qRT-PCR. Subsequently, NSUN2 knockout mice were exploited to build the MCAO model. This study reported that knockout of NSUN2 significantly aggravated brain infarct size and behavioral scores, while reducing 7-day postoperative survival and increasing neuronal apoptosis and injury in MCAO mice. According to the investigation of Western blotting results, decreased PI3K/AKT, ICAM-1 and Bcl-2 protein expressions and increased apoptosis-related protein (Caspase-3/Bax) were found. Overall, this study suggested that NSUN2 may affect cerebral ischemia-reperfusion injury via PI3K/AKT signaling channel and ICAM-1 protein regulation of apoptosis.

Purpose To determine the characteristics of the acute pain after laparoscopic-assisted vaginal hysterectomy (LAVH), laparoscopic myomectomy (LM), and laparoscopic adnexectomy (LA) and compare them with each other. Patients and Methods Patients undergoing LAVH, LM, and LA under general anaesthesia at the First Affiliated Hospital of Wenzhou Medical University between December 2017 and December 2019 were selected. Their data were collected before, during, and after the surgery. We evaluated the degrees of pain in each group of patients and compared them. Results There were differences in the baseline characteristics of the patients in the LAVH, LM, and LA groups. The severity and incidence of postoperative pain were higher in the LAVH group than in the LM and LA groups, followed by the LM and LA groups. Compared with the LA group, the postoperative pain in the LAVH and LM groups was more complicated. The LA group had the lowest incidence of two or more types of moderate to severe pain. The LAVH and LM groups mainly had visceral pain and low back pain, and the LA group mainly had incisional pain. Shoulder pain had the lowest incidence in the three groups. Conclusion There were different postoperative pain characteristics after the LAVH, LM, and LA, and we should clinically adjust analgesia programs for different gynaecological laparoscopic surgeries.