Liangrong Wang

Background/Objectives: Ischemic skeletal muscle undergoes metabolic remodeling that may influence vascular repair. We investigated whether coniferaldehyde (CA) regulates a phosphoglycerate kinase 1 (PGK1)–3-phosphoglycerate (3PG)–L-serine axis in mouse hindlimb ischemia (HLI). Methods: Gastrocnemius tissue from Sham, HLI, and HLI+CA mice (n = 6/group) underwent bulk RNA sequencing and untargeted metabolomics. Public single-cell data and hypoxic human skeletal muscle cells (HSkMCs) were used to examine PGK1. AlphaFold3 modeling, molecular dynamics simulation, and a cellular thermal shift assay (CETSA) were used to evaluate a potential CA–PGK1 interaction. HSkMC-conditioned media were applied to human umbilical vein endothelial cells (HUVECs), with 3PG supplementation where indicated. Results: Amino-acid biosynthesis and glycine/serine/threonine metabolism were enriched in HLI muscle. 3PG increased in HLI versus Sham (p = 0.0170) and decreased in HLI+CA versus HLI (p = 0.0358); L-serine followed the same pattern (p = 0.0274 and p = 0.0428, respectively). Pgk1 expression also increased in HLI versus Sham (p = 0.00348) and decreased in HLI+CA versus HLI (p = 0.00249). Hypoxia increased HSkMC PGK1 together with extracellular 3PG and serine, whereas CA attenuated all three. Modeling predicted a stable CA–PGK1 complex, and CETSA supported a potential CA–PGK1 interaction in cell lysates. Conditioned medium from CA-treated hypoxic HSkMCs improved HUVEC viability, redox, tube-formation, and wound-closure readouts, reduced reactive oxygen species, and increased glutathione peroxidase 4 (GPX4) and solute carrier, family 7 member 11 (SLC7A11); 3PG supplementation reversed these effects. Conclusions: CA modulates a PGK1-linked 3PG/L-serine metabolic axis, while 3PG exposure altered endothelial phenotypes in the conditioned-medium model.

Purpose: Postoperative sore throat (POST) is highly prevalent following double-lumen endobronchial tube (DLT) intubation and negatively impacts recovery. Cheek acupuncture, a micro-needle therapy, provides targeted analgesia through specific acupoint stimulation. This study aimed to evaluate the efficacy of cheek acupuncture in mitigating POST in patients undergoing thoracoscopic lung surgery with DLT intubation. Patients and Methods: Eighty-eight patients were equally randomized to receive either cheek acupuncture (CA group) at bilateral cervical, upper energizer, and upper cervical points for 30 minutes before surgery completion, or sham acupuncture at identical acupoints (CT group). The primary outcome was overall POST incidence within postoperative 48 hours, and secondary outcomes included POST severity, wound pain, rescue analgesic consumption, postoperative nausea and vomiting (PONV) incidence, time to first flatus, postoperative pulmonary complications, adverse events, satisfaction scores, and hospitalization duration. Results: The overall 48-hour postoperative POST incidence was 34.1% in the CA group versus 65.9% in the CT group (risk ratio: 0.517, 95% confidential interval: 0.326 to 0.821, P = 0.003), with the most pronounced reduction observed during the 0– 30 min interval (P < 0.05). Moreover, POST severity was significantly reduced in the CA group at 30 min postoperatively (P < 0.05). Cheek acupuncture was also associated with reduced early wound pain, rescue analgesic requirement, 48-hour PONV incidence, and improved overall anesthesia satisfaction (P < 0.05), and no significant intergroup difference in time to first flatus or hospitalization duration was found (P > 0.05). Conclusion: A single intraoperative session of cheek acupuncture reduces the incidence of early POST after DLT intubation for thoracoscopic lung surgery, with a favorable safety profile.

Purpose: Patients undergoing transurethral prostate surgery often experience catheter-related bladder discomfort (CRBD), which can significantly impairs postoperative recovery. The anatomical convergence between the ilioinguinal and iliohypogastric (II-IH) nerve pathways (T12–L2) and the bladder innervation provides a rationale for evaluating the II-IH nerve block as a potential treatment. This study aimed to investigate the efficacy of ultrasound-guided II-IH nerve block versus intravenous tramadol in reducing postoperative CRBD. Methods: Ninety-four patients undergoing transurethral prostate surgery were equally randomized to receive either 1.5 mg/kg intravenous tramadol or bilateral ultrasound-guided II-IH nerve block with 15 mL of 0.375% ropivacaine per side before extubation. Intravenous tramadol was used as rescue medication for moderate-to-severe CRBD. The primary outcome was the overall incidence of CRBD within postoperative 24 h. Secondary outcomes included CRBD severity, pain scores, adverse events, patient satisfaction, hospitalization duration, and International Prostate Symptom Scores (IPSSs). Results: The overall incidence of CRBD within postoperative 24 h was 56.5% in the II-IH block group versus 65.2% in the tramadol group, with an absolute risk difference of −8.70% (95% confidential interval: −24.71% to 10.92%; p = 0.393). Secondary outcomes, including the incidence of moderate-to-severe CRBD, pain scores at all assessed timepoints, rescue tramadol use, length of hospitalization, and IPSSs were comparable between the two groups (all p > 0.05). However, the II-IH block group demonstrated significantly lower incidences of adverse events (including nausea and vomiting, headache, dizziness, and dry mouth) and a higher proportion of extremely satisfied patients (all p < 0.05). Importantly, no nerve block-related complications were observed in the II-IH block group. Conclusions: Ultrasound-guided II-IH nerve block appeared to be a comparable alternative to intravenous tramadol for reducing CRBD after transurethral prostate surgery, while offering a more favorable safety profile and higher patient satisfaction. Trial registration: Chinese Clinical Trial Registry, ChiCTR2300078552. Registered 12 December 2023, https://www.chictr.org.cn/showproj.html?proj=209712

Background: Flow-mediated dilation (FMD) is commonly used as a diagnostic tool to assess endothelial function, and compared with other methods for stimulating radial artery dilation, FMD offers several advantages such as non-invasiveness, ease of execution, minimal equipment requirements, and negligible risk. The study aimed to investigate the effect of FMD in facilitating radial arterial cannulation in the context of intravenous general anesthesia. Methods: Eighty patients undergoing intravenous general anesthesia and requiring radial artery cannulation were randomized 1:1 to the FMD group and control group. Patients in the FMD group received an upper arm occlusion for 5 min after anesthesia induction, and the cuff was placed without inflation for the equivalent duration in the control group. The primary outcome was first-attempt success rate. Secondary outcomes were the diameter and percentage of dilation of radial artery, overall success rate, total number of attempts, cannulation time, and occurrence of procedure-related complications. Results: Intravenous anesthetic agents significantly dilated the radial artery ( p < 0.05), which was further increased by FMD. An increase in both the first-attempt and overall success rate of radial artery cannulation was demonstrated with the use of FMD (67.5% vs 42.5%, p < 0.05). The total number of attempts needed to cannulate the radial artery was reduced in the FMD group as compared with the control group ( p < 0.05), but no differences in cannulation time and procedure-related complications were found between the two groups ( p > 0.05). Conclusions: FMD induced by a 5-min upper arm occlusion may facilitate radial artery cannulation in patients undergoing intravenous general anesthesia.

ObjectiveProtein kinase C (PKC) has been implicated in the increased contraction of human airway smooth muscle cells (HASMCs) in asthma. Using the three-dimensional collagen gel contraction system, the study aimed to determine the effects of LY333531, a specific inhibitor of the PKC-beta isoform, on the contraction of tumor necrosis factor (TNF)-alpha-sensitized HASMCs.MethodsCultured HASMCs were divided into five groups: the control group received no treatment, and the cells in the TNF-alpha group were sensitized with 10 ng/mL TNF-alpha for 48 h, while TNF-alpha was administered to sensitize HASMCs in the presence of 0.1, 0.2, and 0.5 mu M LY333531 for 48 h in the 0.1LY, 0.2LY, and 0.5LY groups, respectively. Following this, HASMCs contraction was stimulated with 1 mM acetylcholine (ACh) for up to 24 h in each group and assessed using a three-dimensional collagen gel contraction assay. Furthermore, western blot and immunofluorescence analysis were performed.ResultsThe collagen gel contraction assay revealed that TNF-alpha increased the protein expression of phosphorylated PKC-beta 2, CPI-17, and MLC while exacerbating ACh-induced HASMCs contraction. LY333531 significantly attenuated HASMCs contraction and downregulated the protein expression of both p-CPI-17 and p-MLC.ConclusionsAt least in part by regulating CPI-17 and MLC phosphorylation, LY333531 attenuates augmented contraction of TNF-alpha-sensitized HASMCs in a collagen gel contraction system.

Background: Cough is one of the most common complications following intravenous administration of sufentanil during anesthesia induction. The study aimed to investigate the protective effect of alfentanil, afentanyl derivative with short onset time and short duration, in reducing sufentanil-induced cough. Patients and methods: Eighty patients that scheduled for thyroid surgery under general anesthesia were randomly divided into the alfentanil group and normal saline group, with 40 cases per group. Patients in the alfentanil group received intravenous administration of 2 μg/kg alfentanil prior to sufentanil injection during general anesthesia induction, while the same dose of normal saline was administered in the normal saline group. The outcomes measures included the incidence and severity of cough and common side effects of opioids following the administration of sufentanil during the induction of general anesthesia, intraoperative hemodynamics parameters and major adverse events during anesthesia recovery period. Results: The incidence of cough within one minute after the injection of sufentanil during anesthesia induction was 40% in the normal saline group, and the pretreatment of alfentanil significantly reduced the incidence of sufentanil-induced cough to 5% (p < 0.05). Correspondingly, the patients in the alfentanil group had decreased severity of sufentanil-induced cough compared with the normal saline group (p < 0.05). No significant differences in the incidences of common side effects of opioids (dizziness, nausea and vomiting, chest tightness and respiratory depression) within one minute after sufentanil injection were found (p > 0.05). Furthermore, there were no significant differences between the two groups in intraoperative hemodynamic parameters, extubation time, or the incidences of emergence agitation, respiratory depression, delayed recovery from anesthesia and postoperative nausea and vomiting during Postanesthesia Care Unit stay (p > 0.05). Conclusion: Pretreatment with low-dose alfentanil (2 μg/kg) effectively and safely reduced both the incidence and severity of sufentanil-induced cough during anesthesia induction. Clinical Trial Registration Number: Chinese Clinical Trial Registry (identifier: ChiCTR2300069286).

Iron accumulation, which is controlled by transferrin receptor 1 (TfR1), modulates hypoxia-inducible factor-1α (HIF-1α) activation and angiogenesis of hypoxic endothelial cells. The study examined the role of protein interacting with C-kinase 1 (PICK1), a scaffold protein containing PDZ domain, in regulating glycolysis and angiogenesis of hypoxic vascular endothelial cells through its potential effect on TfR1, which features a supersecondary structure that interacts with the PDZ domain. Iron chelator deferoxamine and TfR1 siRNA were employed to assess the impact of iron accumulation on angiogenesis, while the effects of PICK1 siRNA and overexpressing lentivirus on TfR1-mediated iron accumulation were also investigated in hypoxic human umbilical vein vascular endothelial cells (HUVECs). The study found that 72-h hypoxia impaired the proliferation, migration, and tube formation of HUVECs, and reduced the upregulation of vascular endothelial growth factor, HIF-1α, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3, and PICK1, while increasing the expression of TfR1 as compared to 24-h hypoxia. Administration of deferoxamine or TfR1 siRNA reversed these effects and led to increased glycolysis, ATP content, and phosphofructokinase activity, along with increased PICK1 expression. PICK1 overexpression improved glycolysis, enhanced angiogenic capacity, and attenuated TfR1 protein upregulation in hypoxic HUVECs, with higher expression of angiogenic markers, which could be significantly reversed by the PDZ domain inhibitor. PICK1 knockdown exerted opposite effects. The study concluded that PICK1 modulated intracellular iron homeostasis, thereby promoting glycolysis and angiogenesis of HUVECs in response to prolonged hypoxia, at least in part, by regulating TfR1 expression.

Background: This study aimed to establish a clinically relevant animal model for peripheral arterial disease (PAD) that better replicates the complexity observed in human patients. Materials and Methods: Thirty male rats were randomly assigned into the sham (SM), femoral artery resection (FE), constrictor-induced ischemia (CI), two-stage ischemia (TS), or diabetic two-stage ischemia (DT) groups. In the FE group, rats underwent femoral artery resection, whereas the SM group had sham surgery. The CI group received progressive ischemia using two ameroid constrictors, and the TS and DT groups underwent a two-stage ischemia procedure involving initial gradual narrowing with two ameroid constrictors and subsequent femoral artery resection in healthy and diabetic rats, respectively. Perfusion evaluation and functional assessment were conducted at postoperative days 14, 28, and 42. On day 42, hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) protein expression were measured, along with histological examination and immunofluorescence analysis. Results: Motor function deficits and reduced limb reperfusion were most prominent in the TS and DT groups on days 28 and 42 (P < 0.05), exacerbated by type 2 diabetes. Gastrocnemius exhibited upregulated HIF-1α and VEGF protein expression, as well as increased capillary density in response to ischemia. However, the DT group showed significantly lower protein expression and capillary density, along with more severe structural damage compared to other groups (P < 0.05). Conclusion: A clinically relevant rat model of PAD was established by implementing a two-stage ischemia procedure involving initial progressive narrowing and subsequent femoral artery excision in the context of diabetes.

Background: Receptor interacting serine/threonine kinase 1 (RIPK1) mediates apoptosis by regulating the classic proapoptotic effectors Bcl-2-associated X protein (Bax) and Bcl-2 homologous antagonist/killer (Bak). Although Bcl-2-related ovarian killer (Bok) is structurally similar to Bak and Bax, it is unclear whether it mediates apoptosis in skeletal muscle ischemia reperfusion (IR) injury. We hypothesized that by regulating Bok-mediated apoptosis, inhibiting RIPK1 with necrostatin-1 would reduce skeletal muscle IR injury. Methods: Rats were randomized into four groups: sham (SM), IR, IR treated with necrostatin-1 (NI), or vehicle dimethyl sulfoxide (DI). For the IR group, the right femoral artery was clamped for 4 hours and then reperfused for 4 hours, and for the NI and DI groups, necrostatin-1 (1.65 mg/kg) and the equal volume of dimethyl sulfoxide were intraperitoneally administered prior to IR induction. The structural damage of muscle tissue and protein expression of Bok, Bcl-2, and cleaved caspase-3 were investigated, and apoptotic cells were identified with terminal dUTP nick-end labeling (TUNEL) staining. In vitro, human skeletal muscle cells (HSMCs) were exposed to 6 hours of oxygen-glucose deprivation followed by normoxia for 6 hours to establish an oxygen-glucose deprivation/reoxygenation (OGD/R) model. To determine the role of Bok, cell viability, lactate dehydrogenase (LDH) release, and flow cytometry were examined to demonstrate the effects of necrostatin-1 and Bok knockdown on the OGD/R insult of HSMCs. Results: Necrostatin-1 pretreatment markedly reduced IR-induced muscle damage and RIPK1, Bok, and cleaved caspase-3 expression, whereas upregualted Bcl-2 expression (p < 0.05). Furthermore, necrostatin-1 prevented mitochondrial damage and decreased TUNEL-positive muscle cells (p < 0.05). In vitro, HSMCs treated with necrostatin-1 showed reduced Bok expression, increased cell viability, and reduced LDH release in response to OGD/R (p < 0.05), and Bok knockdown significantly blunted the OGD/R insult in HSMCs. Conclusion: Necrostatin-1 prevents skeletal muscle from IR injury by regulating Bok-mediated apoptosis.