Xuliang Huang’s research while affiliated with First Affiliated Hospital of China Medical University and other places

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.

Dichloroacetate (DCA), a pyruvate dehydrogenase kinase inhibitor, is often used to treat lactic acidosis and malignant tumors. Increasing studies have shown that DCA has neuroprotective effects. Here, we explored the role and mechanism of DCA in Sepsis associated encephalopathy (SAE). Single-cell analysis was used to determine the important role of PDK4 in SAE and identify the cell type. GO and GSEA analysis were used to determine the correlation between DCA and pyroptosis. Through LPS + ATP stimulation, a microglia pyroptosis model was established to observe the expression level of intracellular pyroptosis-related proteins under DCA intervention, and further detect the changes in intracellular ROS and JC-1. Additionally, a co-culture environment of microglia and neuron was simply constructed to evaluate the effect of DCA on activated microglia-mediated neuronal apoptosis. Finally, Novel object recognition test and the Morris water maze were used to explore the effect of DCA on cognitive function in mice from different groups after intervention. Based on the above experiments, this study concludes that DCA can improve the ratio of peripheral and central M1 macrophages, inhibit NLRP3-mediated pyroptosis through ROS and mitochondrial membrane potential (MMP). DCA can reduce neuron death caused by SAE and improve cognitive function in LPS mice. In SAE, DCA may be a potential candidate drug for the treatment of microglia-mediated neuroinflammation. Graphical Abstract

Background Neuroinflammation mediated by microglial pyroptosis is an important pathogenic mechanism of septic encephalopathy (SAE). It has been reported that TRIM45 is associated with tumours and inflammatory diseases. However, the role of TRIM45 in SAE and the relationship between TRIM45 and microglial pyroptosis are unknown. In this study, we found that TRIM45 played an important role in regulating microglial pyroptosis and the molecular mechanism. Methods SAE was induced by intraperitoneal injection of LPS in WT and AAV-shTRIM45 mice. BV2 cells were treated with LPS/ATP in vitro. Cognitive function was assessed by the Morris water maze. Nissl staining was used to evaluate histological and structural lesions. ELISA was used to dectect neuroinflammation. qPCR was used to detect the mRNA levels of inflammatory cytokines, NLRP3, and autophagy genes. Western blotting and immunofluorescence analysis were used to analyse the expression of the proteins. Changes in reactive oxygen species (ROS) in cells were observed by flow cytometry. Changes in mitochondrial membrane potential in BV2 cells were detected by JC-1 staining. Peripheral blood mononuclear cells were extracted from blood by density gradient centrifugation and then used for qPCR, western blotting and flow detection. To further explore the mechanism, we used the overexpression plasmids TRIM45 and Atg5 as well as siRNA-TRIM45 and siRNA-Atg5 to analyse the downstream pathway of NLRP3. The protein and mRNA levels of TRIM45 in peripheral blood mononuclear cells from sepsis patients were examined. Results Knocking down TRIM45 protected against neuronal damage and cognitive impairment in septic mice. TRIM45 knockdown inhibited microglial pyroptosis and the secretion of inflammatory cytokines in vivo and in vitro, which was mediated by NLRP3/Gsdmd-N activation. Overexpression of TRIM45 could activate NLRP3 and downstream proteins. Further examination showed that TRIM45 regulated the activation of NLRP3 by altering Atg5 and regulating autophagic flux. It was also found that overexpression and knockdown of TRIM45 affected the changes in ROS and mitochondrial membrane potential. Thus, knocking down TRIM45 could reduce microglial pyroptosis, the secretion of proinflammatory cytokines, and neuronal damage and improve cognitive function. In addition, the level of TRIM45 protein in septic patients was increased. There was a positive linear correlation between APACHE II score and TRIM45, between SOFA score and TRIM45. Compared to group GCS > 9, level of TRIM45 were increased in group GCS ≤ 8. Conclusion TRIM45 plays a key role in neuroinflammation caused by LPS, and the mechanism may involve TRIM45-mediated exacerbation of microglial pyroptosis via the Atg5/NLRP3 axis. Graphical Abstract

Background We studied AKI incidence and prognosis in cardiac surgery patients under and over 60 years old. Methods We studied AKI in patients who underwent cardiac surgery at the First Affiliated Hospital of Wenzhou Medical University between Jan 2020 and Dec 2021, using improved global prognostic criteria for diagnosis. Results After analyzing 781 patients (402 males, 379 females), AKI incidence after surgery was 30.22 %. Adjusting for propensity scores revealed no significant difference in AKI incidence between young males (24.1 %) and females (19.3 %). However, young females had higher AKI stages. Among older patients, AKI incidence was comparable between males (43.4 %) and females (42.2 %), but females had longer intubation times. Independent risk factors for AKI included age, male gender, and BMI, while intraoperative hemoglobin level was protective. Conclusions No gender gap in AKI frequency for <60 years old and ≥60 years old post-cardiac surgery, yet women display increased AKI severity and extended intubation duration.

Background Neuroinflammation mediated by microglia pyroptosis activation is an important pathogenesis of septic encephalopathy(SAE). It is reported that TRIM45 is associated with tumors and inflammatory diseases. However, the role of TRIM45 in SAE and the relationship between TRIM45 and microglia pyroptosis are unknown. In this study, we found that TRIM45 plays an important role in regulating microglia pyroptosis and its molecular mechanism. Methods SAE was induced by lps in mice transfected with adenovirus shTRIM45. BV2 was treated with lps + ATP in vitro. Cognitive function was assessed by Morris water maze. Nissl staining and Tunnel staining were used to evaluate histological and structural lesions. QPCR was used to detect mRNA level of inflammatory cytokines, NLRP3, autophagy proteins. Western blotting and immunofluorescence were used to analyze the expression of the proteins. The changes of ROS in cells were observed by flow cytometry, The changes of mitochondrial membrane potential in BV2 cells were detected by JC-1 staining. Peripheral blood mononuclear cells were extracted from blood by density gradient centrifugation and then used for QPCR and Western blotting analysis. In order to further explore its mechanism, we used overexpression plasmids TRIM45 and Atg5, siRNA-TRIM45 to analyze the downstream pathway of NLRP3. The protein and mRNA of TRIM45 in the peripheral blood mononuclear cells from sepsis patients were detected. Results Knocking down TRIM45 could protect the neuron damage and cognitive impairment of septic mice. TRIM45 knockdown can inhibit microglia pyroptosis and the secretion of inflammatory cytokines in vivo and in vitro, which is mediated by activating NLRP3/Gsdmd-N. Overexpression TRIM45 can activate NLRP3 and downstream proteins. Further exploration found that TRIM45 regulated the activation of NLRP3 by changing Atg5 protein and regulating autophagy flux. It was also found that overexpression and knockdown of TRIM45 would affect the changes of ROS and mitochondrial membrane potential. In short, knocking down TRIM45 can reduce microglia pyroptosis, reduce the secretion of pro-inflammatory cytokines, reduce neuronal damage and improve cognitive function. The levels of TRIM45 mRNA and protein in septic patients were increased, and TRIM45 mRNA level of peripheral blood monocytes and APACHE II score were positive linear correlation in some patients with sepsis. Conclusion TRIM45 plays a key role in neuroinflammation caused by lps, and the possible mechanism is that TRIM45 aggravates microglia pyroptosis via Atg5/NLRP3 axis.

Sepsis-induced cardiomyopathy (SIC) is one of the most common complications of infection-induced sepsis. An imbalance in inflammatory mediators is the main factor leading to SIC. N6-methyladenosine (m6A) is closely related to the occurrence and development of sepsis. m6A reader YTH Domain Containing 1 (YTHDC1) is an m6A N6-methyladenosine recognition protein. However, the role of YTHDC1 in SIC remains unclear. Herein, we demonstrated that YTHDC1-shRNA inhibits inflammation, reduces inflammatory mediators, and improves cardiac function in a lipopolysaccharide (LPS)-induced SIC mouse model. Based on the Gene Expression Omnibus (GEO) database analysis, serine protease inhibitor A3N (SERPINA3N) is a differential gene of SIC. Further, RNA immunoprecipitation (RIP) indicated that SERPINA3N mRNA can bind to YTHDC1, which regulates the expression of SERPINA3N. SERPINA3N-siRNA reduced LPS-induced inflammation of cardiac myocytes. In conclusion, the m6A reader YTHDC1 regulates SERPINA3N mRNA expression to mediate the levels of inflammation in SIC. Such findings add to the relationship between m6A reader YTHDC1 and SIC, providing a new research avenue for the therapeutic mechanism of SIC.

Background: Postoperative nausea and vomiting (PONV) is one of the most common complications after general anesthesia. The traditional comprehensive management of PONV usually uses one or two drugs, but this regimen fails to meet the requirements of the latest version of PONV guidelines. The purpose of this study was to evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on high-risk PONV patients who are undergoing laparoscopic gynecological surgery. Methods: In total, 162 high-risk PONV patients were randomly divided into an experimental group (n = 81) and a control group (n = 81). Both groups were injected with 4 mg of dexamethasone and 0.25 mg of palonosetron. In the experimental group, Nei-guan (PC6) and He-gu (LI4) were stimulated by a transcutaneous acupoint electrical stimulation instrument (HANS200E) 30 min before the surgery. The control group also received electrodes but no stimulation. Variance analysis and rank sum test were used to compare the differences between the two groups. Results: The results of the incidence of postoperative nausea, vomiting, NRS score, degree of abdominal distension, and time to first flatus in the experimental group were lower than those in the control group. Nursing satisfaction of the experimental group was higher than that of the control group. Conclusions: The study demonstrates that TEAS combined with dexamethasone and palonosetron can effectively prevent PONV, reduce postoperative abdominal distension and postoperative pain, and shorten the first postoperative flatus time in high-risk patients with PONV. At the same time, it can improve nursing satisfaction.

Clinically, ischemic stroke is a serious disease hard to cure, therefore it is of great importance to slow down the depletion of ATP to enhance the tolerance of ischemic tissue through aggressive preconditioning. Electroacupuncture (EA) preconditioning has been reported to produce a tolerance for cerebral ischemia, with the underlying mechanism remaining unclear. Given that the P2X7 receptor (P2X7R) mediates the stimulation of microglial cells and is involved in the developmental process of cerebrum ischemia-reperfusion (I/R) damage, we hypothesized that the protective effect of Electroacupuncture preconditioning might be associated with the downregulated P2X7R. In this study, 2 '(3')-O-(4-benzoyl) adenosine triphosphate (BzATP) was utilized as a P2X7R agonist in rats with cerebral ischemia-reperfusion injury (MCAO), from which we found that EA preconditioning ameliorated neurologic scoring, decreased infarction volume, and necrotic neurons, and decreased the release of cytokines, while BzATP exacerbated cerebrum I/R damage and inflammation events compared with the favorable efficacy of EA. In addition, BzATP reversed the positive effects mediated by EA, which was consistent with increased expression of P2X7R. These findings suggest that EA induces cerebral ischemic tolerance to ischemia-reperfusion(I/R) damage by suppressing the expression of P2X7R and the release of inflammatory factors.