Small extracellular vesicles have been widely studied for their therapeutic properties and ability to deliver bioactive molecules. In addition to secretory vesicles, cells contain small intracellular vesicles involved in physiological and metabolic processes, whose therapeutic potential remains unexplored. Here we developed protocols to isolate small intracellular vesicles from multiple cell types and systematically compared their molecular and functional profiles to extracellular vesicles. Intracellular vesicles are smaller, yield higher quantities and demonstrate enhanced cellular uptake in both in vitro and in vivo models. Molecular profiling revealed that intracellular vesicles are enriched in proteins associated with the endoplasmic reticulum and Golgi apparatus, possess distinct microRNA signatures linked to intracellular membrane systems, and contain elevated levels of phospholipids such as phosphatidylcholine and phosphatidylethanolamine. Vesicles derived from umbilical cord mesenchymal stem cells showed superior therapeutic efficacy in a model of retinal degeneration by reducing endoplasmic reticulum stress and delivering neuroprotective factors. In addition, intracellular vesicles exhibited enhanced drug-loading capacity and efficient delivery of lipophilic compounds to the retina. These findings position intracellular vesicles as promising candidates for therapeutic applications. A protocol to isolate small intracellular vesicles from human cells is reported; these are molecularly distinct from small extracellular vesicles and show superior therapeutic potential in retinal degeneration and drug delivery.

Background Diabetic retinopathy (DR) is the leading cause of vision loss in the working-age population, traditionally attributed to metabolic disorders and microvascular damage induced by chronic hyperglycemia. Emerging evidence indicates that cellular senescence acts as a critical upstream driver of DR, with senescent cells accumulating in the retina and secreting a senescence-associated secretory phenotype (SASP) that promotes chronic inflammation, pathological angiogenesis, and disruption of the blood-retinal barrier (BRB). Main text This review adopts a neurovascular unit (NVU) perspective to synthesize cell-type-specific senescence phenotypes in the diabetic retina, emphasizing that endothelial cell, pericyte, retinal pigment epithelium, microglial, and retinal ganglion cell senescence constitute an interconnected pathological network rather than isolated events. NVU-wide senescence establishes a self-reinforcing cycle: senescent endothelial and pericyte dysfunction drives vascular instability; retinal pigment epithelial cells (RPE) senescence compromises the outer BRB; and glial and retinal ganglion cell (RGC) senescence amplifies neuroinflammation and neurodegeneration. We dissect key molecular drivers—oxidative DNA damage, epigenetic reprogramming (miRNA/Sirtuins), mitophagy impairment, and SASP-mediated inflammatory feedforward—and critically evaluate emerging senotherapeutics (senolytics, senomorphics, and precision delivery platforms) within an evidence-graded framework. Conclusions Cellular senescence establishes a progressive senescence-inflammation vicious cycle in DR, affecting multiple interconnected cell types within the NVU and causing neurovascular dysfunction. Targeting cellular senescence through senescent cell clearance, SASP inhibition, mitochondrial homeostasis regulation, and epigenetic modulation offers promising new therapeutic directions. With the advancement of novel senotherapeutics and precision technologies, personalized early treatment and retinal repair for DR may become achievable in the future.

Point-of-care test (POCT) of blood viscoelasticity can reflect the coagulation status of patients accurately and timely. POCT can be used to monitor the effect of preoperative antithrombotic drugs on coagulation function, which will inform the selection of appropriate surgical timing. It can also be applied to analyze the reasons of intraoperative bleeding and guide the transfusion of blood products. Also, it is useful to assess the risk of postoperative thromboembolism and hint the need for prophylactic anticoagulation. This article mainly introduces the principles and clinical application of several frequently used POCTs for blood viscoelasticity, with a focus on their role in special types of diseases in which coagulation function changes significantly, such as cardiac disease, trauma, pathological obstetrics, and liver disease. Furthermore, we describe the role of microparticle in coagulation, which is a novel potential biomarker for diagnosing thrombotic disorders and possesses potential to be applied in POCTs of blood viscoelasticity.

The study investigates the effectiveness of immersive virtual reality (VR) as a nonpharmaceutical approach to manage postoperative pain in patients following thoracoscopic surgery. In this single-center, triple-arm pilot randomized controlled trial (RCT), 61 postsurgical patients with a postoperative pain numerical rating scale (NRS) score ≥4 after receiving standard analgesia were included and assigned to either a quantum clinics-VR (QTC-VR) group, a Placebo-VR group, or a control group. The QTC-VR group engaged in a daily 10-minute interactive pain relief 3D-VR program, while the Placebo-VR group watched a daily 10-minute relaxation-based 2D film through VR headsets for three days following surgery. 61 postsurgical patients were randomized and allocated (21 in the QTC-VR group, 20 in the Placebo-VR group, and 20 in the control group) in the final intention-to-treat (ITT) analyses. Compared with patients receiving Placebo-VR intervention, patients reported significantly lower pain scores following the daily QTC-VR intervention on postoperative days 1 (mean difference, −0.889; 95% CI, −1.464 to −0.314; P < 0.001), 2 (mean difference, −0.631; 95% CI, −1.211 to −0.051; P = 0.014), and 3 (mean difference, −0.798; 95% CI, −1.345 to −0.251; P < 0.001), respectively. Additionally, patients receiving QTC-VR intervention also reported high satisfaction and tolerable adverse events with their treatment. In conclusion, this pilot RCT demonstrates that QTC-VR might be a promising intervention for pain management post-thoracoscopic surgery, warranting further validation in ongoing phase III trials.

Corneal neovascularization (CNV) is a significant risk factor for visual impairment. The efficiency and side effects of current CNV treatments, such as steroids and antivascular endothelial growth factor agents, are still debated. In addition, the bioavailability of topical drugs is usually hindered by tears, blinking, and the corneal anatomy. Therefore, finding a new therapeutic strategy is important. This study aimed to examine the function of the new therapeutic agent capmatinib (Cap), a highly selective inhibitor of MET that plays an important role in angiogenesis, in treating CNV. In this study, we first investigated the role of the HGF/c-MET axis in CNV and the therapeutic effect of Cap in a corneal alkali burn model. We synthesised a genipin-crosslinked gelatine-based hydrogel containing Cap (Cap-Gel). We observed a more significant therapeutic effect with the Cap-Gel than with Cap alone, as well as the alleviation of inflammatory infiltration and fibrosis. On day 14, the Cap-Gel group showed the most significant inhibition of corneal neovascularization, with the shortest neovessel length (0.48 ± 0.13 mm), smallest CNV area (3.77 ± 0.78 mm2), and lowest clinical assessment score (3.33 ± 0.52). Taken together, our results suggest that Cap-Gel could be a promising drug candidate for treating CNV.

Oxygen-induced retinopathy (OIR) mouse model is widely used to study retinal neovascular diseases. Although the OIR procedure has been well established in detail, few studies to date have examined the effect of intravitreal injection using different-sized syringe needles at different time intervals after mouse pups returned to room air on this model. Initially, the significant reduction of NV and VO areas in the vehicle-controls of OIR drew our attention. We found that intravitreal injection performed using a 33 g-needle at 2 h after the pups returned to room air resulted in minimal NV and VO areas, causing a failure of OIR model. The results of ERG and OCT testing showed that 34 g-needle was more suitable than a 33 g-needle for intravitreal injection in the OIR model. We then investigated the effect of time interval after pups returned to room air on the OIR model. The results indicated that 8-24 h was a more suitable time for performing intravitreal injection. In conclusion, appropriate control of the effects of intravitreal injection on OIR requires attention to gauge of needle used, and the time interval after return of pups to room air.

Several common retinal diseases that cause blindness are characterised by pathological neovascularisation accompanied by inflammation and neurodegeneration, including retinopathy of prematurity (ROP), diabetic retinopathy (DR), age-related macular degeneration (AMD), and retinal vein occlusion (RVO). The current treatment strategies for these diseases have limited benefits. Thus, safer and more effective alternative approaches are required. In this study, we loaded small extracellular vesicles (sEVs) derived from mesenchymal stem cell (MSC) with pigment epithelium-derived factor (PEDF), and tested the therapeutic effect of PEDF-loaded sEVs (PEDF-sEVs) using an oxygen induced retinopathy (OIR) mouse model, aiming to establish a new therapy strategy for the treatment of retinal pathological angiogenesis. We formulated PEDF-loaded sEVs (PEDF-sEVs) containing high concentrations of PEDF and evaluated their effects through in vivo and in vitro experiments. In OIR mice, PEDF-sEVs showed significantly better effects on retinal avascular areas, inflammation, and neuronal degeneration compared with the anti-vascular endothelial growth factor (VEGF) drug, which may indicate a possible advantage of PEDF-sEVs over anti-VEGF drugs in the treatment of pathological neovascularisation. In vitro, PEDF-sEVs greatly inhibited endothelial cell (EC) proliferation, migration, and tube formation by suppressing the VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK) and AKT (also known as Protein Kinase B). All experiments and analyses were performed in triplicate. PEDF-sEVs were more effective than PEDF or sEVs alone, both in vitro and in vivo. Furthermore, to determine the distribution of PEDF-sEVs, we used DiD-labelled sEVs and FITC-labelled PEDF to track the sEVs and PEDF, respectively. We found that PEDF-sEVs effectively reduced the degradation of PEDF. Loading PEDF on sEVs effectively enhanced the anti-angiogenic, anti-inflammatory, and neuroprotective effects of PEDF by increasing the stability and penetrability. These results suggest a potential role for PEDF-sEVs in retinal pathological neovascularisation.

Background: Stroke is a major health concern and a leading cause of mortality and morbidity. We and other groups have documented that hyperbaric oxygen preconditioning could significantly alleviate neuronal damage in ischemia‒reperfusion models through various mechanisms. However, we found that some of the subjects did not benefit from preconditioning with hyperbaric oxygen. The preconditioning phenomenon is similar to vaccination, in which the endogenous survival system is activated to fight against further injuries. However, with vaccine inoculations, we could test for specific antibodies against the pathogens to determine if the vaccination was successful. Likewise, this experiment was carried out to explore a biomarker that can reveal the effectiveness of the preconditioning before neuronal injury occurs. Methods: Middle cerebral artery occlusion (MCAO) was used to induce focal cerebral ischemia-reperfusion injury. 2D-DIGE-MALDI-TOF-MS/MS proteomic technique was employed to screen the differentially expressed proteins in the serum of rats among the control (Con) group (MCAO model without hyperbaric oxygen (HBO) preconditioning), hyperbaric oxygen protective (HBOP) group (in which the infarct volume decreased after HBO preconditioning vs. Con), and hyperbaric oxygen nonprotective (HBOU) group (in which the infarct volume remained the same or even larger after HBO preconditioning vs. Con). Candidate biomarkers were confirmed by western blot and enzyme linked immunosorbent assay (ELISA), and the relationship between the biomarkers and the prognosis of cerebral injury was further validated. Results: Among the 15 differentially expressed protein spots detected in the HBOP group by Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), 3 spots corresponding to 3 different proteins (haptoglobin, serum albumin, and haemopexin) products were identified by MALDI-TOF-MS/MS. Serum albumin and haemopexin were upregulated, and haptoglobin was downregulated in the HBOP group (p < 0.05 vs. Con and HBOU groups). After the western blot study, only the changes in haemopexin were validated and exhibited similar changes in subjects from the HBOP group in accordance with MALDI-TOF-MS/MS proteomic analysis and enzyme linked immunosorbent assay (ELISA) analysis. The serum level of the hemopexin (HPX) at 2 h after HBO preconditioning was correlated with the infarct volume ratio after MCAO. Conclusions: Haemopexin may be developed as a predictive biomarker that indicated the effectiveness of a preconditioning strategy against cerebral ischaemic injury.

Photoreceptors (PRs) degeneration is central to visual impairment and loss in most blind retinal diseases, including age-related macular disease (AMD) and diabetic retinopathy (DR). PRs are susceptible to oxidative stress owing to their unique metabolic features. Accumulating evidence has demonstrated that the targeting oxidative stress is a promising treatment strategy for PR degeneration. Herein, we introduced potent anti oxidative platinum nanoparticles (Pt NPs) to treat PRs degeneration in this study. The Pt NPs exhibited multi enzymatic antioxidant activity and protected PRs from H2O2-induced oxidative damage in vitro assays. Based on the same mechanism, the intravitreal injection of Pt NPs significantly reduced cell apoptosis, maintained retinal structure and preserved retinal function in a mouse model of light-induced retinal degeneration (LIRD). Most importantly, the results of RNA sequencing showed that the transcription of antioxidative genes was upregulated, and metabolic reprogramming occurred in the LIRD-retina after treatment with Pt NPs, both of which benefited retinal survival from oxidative damage. The results indicated that Pt NPs were indeed potent therapeutic candidates for PRs degeneration in blind retinal diseases.

Autoimmune uveitis is a major cause of vision loss and glucocorticoids are major traditional medications, which may induce serious complications. Rapamycin has been demonstrated to exhibit immunosuppressive effects and is promising to be used in treating uveitis by intravitreal injection. However, repeated and frequent intravitreal injections increase the risk of severe ocular complications, while the efficacy of subconjunctival injection of rapamycin is low since it is difficult for rapamycin to penetrate eyeball. Recently, small extracellular vesicles (sEVs) have attracted considerable research interest as natural drug delivery systems that can efficiently cross tissues and biological membranes. SEVs derived from mesenchymal stem cells (MSC-sEVs) also can exert immunosuppressive effect and ameliorate experimental autoimmune uveitis (EAU). The aim of this study was to construct a Rapamycin-loaded MSC-sEVs delivery system (Rapa-sEVs) and investigate its therapeutic effect on EAU by subconjunctival injection. Rapa-sEVs were prepared by sonication and characterized by nanoparticle tracking analysis, transmission electron microscopy, and western blotting. Clinical and histological scores were obtained to assess the treatment efficacy. Additionally, T cell infiltration was evaluated by flow cytometry. The results indicated that Rapa-sEVs could reach the retinal foci after subconjunctival injection. Compared to sEVs and rapamycin alone, Rapa-sEVs can produce a more marked therapeutic effect and reduce ocular inflammatory cell infiltration. Overall, MSC-sEVs have significant potential for the delivery of rapamycin to treat EAU. Subconjunctival injection of Rapa-sEVs may be contender for efficacious steroid-sparing immunomodulatory therapy.

Neuromodulation is becoming more and more important in studying brain function, disease treatment, and brain–computer interfaces. However, traditional regulation methods cannot effectively achieve both wireless regulation and highly sensitive response, which are essential factors in neuromodulation. In this paper, a “magnetism‐optogenetic” system is constructed, which uses a magnetic field to drive mechanoluminescent materials (ZnS:Cu) to generate light, thus stimulating photogenetic proteins. This system effectively combines the wireless magnetic regulation with the high sensitivity of optogenetics. The results show that the luminous intensity of this system changes with the power of an external magnetic field. In addition, under the continuous stimulation of the wireless magnetic field, this system can activate hippocampal‐related neural responses and induce the expression of C‐fos. In the end, this system can further regulate the movement behavior of rats with C1V1 protein expression in the primary motor cortex. This new magnetism‐optogenetic system will provide an excellent reference for wireless and highly sensitive neuromodulation.

N-methyl-D-aspartate (NMDA) receptor activation is known to be critical in remifentanil-induced hyperalgesia. Evidence indicates that iron accumulation participates in NMDA neurotoxicity. This study aims to investigate the role of iron accumulation in remifentanil-induced hyperalgesia. Remifentanil was delivered intravenously in rats to induce hyperalgesia. The NMDA receptor antagonist MK-801 was intrathecally administrated. The levels of divalent metal transporter 1 without iron-responsive element [DMT1(-)IRE] and iron were detected. Behavior testing was performed in DMT1(-)IRE knockdown rats and rats treated with iron chelator DFO. Meanwhile, the spinal dorsal horn neurons were cultured and transfected with DMT1(-)IRE siRNA, and then respectively incubated with remifentanil and MK-801. The levels of intracellular Ca2+ and iron were assessed by fluorescence imaging. Our data revealed that spinal DMT1(-)IRE and iron content significantly increased in remifentanil-treated rats, and MK-801 inhibited the enhancements. DMT1(-)IRE knockdown and DFO prevented against remifentanil-induced hyperalgesia. Notably, the levels of Ca2+ and iron increased in remifentanil-incubated neurons, and these growths can be blocked by MK-801. DMT1(-)IRE knockdown attenuated iron accumulation but did not influence Ca2+ influx. This study suggests that DMT1(-)IRE-mediated iron accumulation is likely to be the downstream event following NMDA receptor activation and Ca2+ influx, contributing to remifentanil-induced hyperalgesia. PERSPECTIVE: Remifentanil-induced hyperalgesia is common even when used within clinical accepted doses. This study presents that aberrant iron accumulation is involved in the development of remifentanil-induced hyperalgesia in vivo and in vitro. Iron chelation may be a potential therapeutic strategy for the prevention of hyperalgesia in populations at high risk.

Early diagnosis and precise monitoring of the development of proliferative diabetic retinopathy (PDR) can significantly improve therapeutic strategies and help decrease blindness caused by it. Extracellular vesicles (EVs) were recently found to be involved in intercellular communications and are a potential source for the discovery of novel biomarkers. The current study aims to investigate the effectiveness of microRNAs (miRNAs) encapsulated in small EVs (sEVs) as minimally invasive biomarkers for PDR. SEVs were extracted from plasma of healthy subjects, diabetic patients, nonPDR patients and PDR patients. Then, we performed microarray analysis to determine the miRNA expression profile. MiR-431-5p expression doubled in the PDR patients compared with the healthy controls and the diabetic patients. We further found that miR-431-5p expression was 2.3 times higher in 4-hydroxynonenal treated human retinal capillary endothelial cells (HRCECs) than the control. After transfection with miR-431-5p mimics, proliferation of HRCECs was promoted, while transfection with miR-431-5p inhibitor demonstrated the opposite effect. The present findings indicate that circulating sEVs showed a differential miRNA profile in PDR patients. MiR-431-5p was involved in the pathogenesis of PDR development and may function as a novel biomarker for PDR.

Tumour revascularization and the consequent radioresistance activated by the up-regulated angiogenic pathway after radiation exposure remain a major bottleneck for improving the tumouricidal effect of radiotherapy (RT) in hepatocellular carcinoma (HCC). Herein, we show that fabricated aminopeptidase N (ANP/CD13)-targeting Gd-hybridized gold nanomolecules (tGd-GNMs) can efficaciously suppress tumour revascularization and the consequent radioresistance, and then synergize in augmenting the RT response. Both in vitro and in vivo experiments demonstrate that the targeted delivery of vascular endothelial growth factor (VEGF) siRNA into the tumour site and the generation of an abundance of intratumourally cytotoxic reactive oxygen species (ROS) under X-ray radiation by the tGd-GNMssiRNA complex has the capability to down-regulate VEGF gene expression and strengthen the radiation response. Furthermore, the tGd-GNMssiRNA complex contributes to excellent active tumour targeting ability, remarkably enhancing tumour contrast in the fluorescence, computed tomography (CT) and magnetic resonance (MR) imaging modalities in real-time with a long imaging time window. Overall, the synthesized tGd-GNMssiRNA complex with excellent potentiation of the antitumour ability and real-time multimodal imaging ability represents a promising visualized theranostic nanoplatform for the treatment of HCC.

BACKGROUND:Remifentanil-induced postoperative hyperalgesia is an intractable side effect of the clinical use of remifentanil, the mechanism of which remains obscure, especially in the peripheral nervous system. N-methyl-D-aspartate receptor (NMDAR) phosphorylation in dorsal root ganglion (DRG) plays a pronociceptive role in neuropathic pain. The contribution of the P2Y1 purinergic receptor (P2Y1R) in DRG to pain hypersensitivity derived from various origins and P2Y1R upregulation-induced NMDAR activation in neurons have also been uncovered. This study aimed to investigate whether P2Y1R participates in nociceptive processing in the DRG and spinal cord in remifentanil-induced postoperative hyperalgesia. METHODS:Rats with remifentanil-induced postoperative hyperalgesia were intrathecally injected with NMDAR antagonist MK801 or P2Y1R antagonist MRS2179 at 10 min prior to remifentanil infusion. Mechanical allodynia, heat hyperalgesia, and cold hyperalgesia were measured at -24 h, 2 h, 6 h, 24 h, and 48 h following remifentanil infusion. The P2Y1R expression and NMDAR expression and phosphorylation in DRG ipsilateral to the incision were detected by Western blot and immunofluorescence. RESULTS:Incision and remifentanil induced mechanical allodynia, heat hyperalgesia, and cold hyperalgesia accompanied by upregulated P2Y1R expression, increased NMDAR subunit NR1 expression and phosphorylation at Ser896, and NR2B expression and phosphorylation at Tyr1472 in DRG. Inhibition of NMDAR phosphorylation by MK801 effectively attenuated remifentanil-induced postoperative hyperalgesia. Furthermore, P2Y1R blockade by MRS2179 not only lessened remifentanil-evoked postoperative hypersensitivity to mechanical, heat, and cold stimuli, but also suppressed the increases in NR1 and NR2B expression and phosphorylation in DRG induced by incision and remifentanil. CONCLUSION:The process by which P2Y1R mediates NMDAR expression and phosphorylation represents a mechanism of remifentanil-induced postoperative hyperalgesia in the DRG and/or spinal cord.

目的 探讨右美托咪定辅助硬膜外麻醉在中转剖宫产手术中的应用价值,观察不同方案的麻醉效果.方法 选取2016年1月-2019年12月在该院阴道试产失败中转剖宫产并接受右美托咪定辅助硬膜外麻醉的42例产妇为观察组,同期在该院阴道试产失败中转剖宫产并接受生理盐水辅助硬膜外麻醉的38例产妇为对照组,回顾性分析两组研究对象临床资料.比较两组产妇感觉阻滞起效时间,感觉阻滞时间及最高阻滞平面;比较两组产妇镇静评分(Ramsay评分),牵拉反应评分及新生儿娩出后1min、5min的Apgar评分;比较两组产妇麻醉前(T0)、切皮时(T1)胎儿取出时(T2)、关腹时(T3)的平均动脉压(MAP)、心率(HR)和血氧饱和度(SPO2)等血流动力学情况;记录两组产妇不良反应发生情况.结果 观察组产妇感觉阻滞起效时间短于对照组,感觉阻滞时间长于对照组,最高阻滞平面高于对照组,差异均有统计学意义(均P＜0.05).观察组产妇Ramsay评分和牵拉反应评分均高于对照组,差异均有统计学意义(均P＜0.05).两组新生儿Apgar评分比较,差异均无统计学意义(均P＞0.05).T1～T3时,观察组产妇HR、MAP水平均低于T0时,且观察组低于对照组,差异均有统计学意义(均P＜0.05).两组产妇各时点SPO2水平相近,差异均无统计学意义(均P＞0.05).两组产妇不良反应发生率比较,差异无统计学意义(P＞0.05).结论 右美托咪定辅助硬膜外麻醉在中转剖宫产手术中的应用效果较好,更利于稳定产妇术中血流动力学稳定,产妇获得的镇静效果及感觉阻滞效果更好,且不会增加不良反应,安全性较高.

Small extracellular vesicles (sEVs) derived from the plasma have been increasingly recognized as important vehicles of intercellular communication and potential sources of new biomarkers for multiple diseases. In this study, proteomic profiles of plasma sEVs from normal subjects and diabetic patients with or without diabetic retinopathy (DR) were systematically compared using iTRAQ-based quantitative proteomics. Among a total of 901 identified proteins in plasma sEVs (false discovery rate (FDR) < 1%), 90 proteins were found to have significantly changed levels in DR. Based on the findings from the proteomic analysis, the role of tumor necrosis factor-α-induced protein 8 (TNFAIP8) in promoting human retinal microvascular endothelial cell (HRMEC) proliferation was investigated. The enzyme-linked immunosorbent assay (ELISA) showed that TNFAIP8 levels in plasma sEVs and vitreous are elevated in DR, whereas not statistically different in large EVs (lEVs) and plasma. In addition, in vitro experiments demonstrated that 4-hydroxynonenal (4-HNE) increased the expression of TNFAIP8 in HRMECs. TNFAIP8 significantly increased HRMECs cell viability and promote cell migration and tube formation, and the depletion of TNFAIP8 impaired HRMEC proliferation. We demonstrated that TNFAIP8 in plasma sEVs could be used as a potential biomarker of DR. Functional studies suggested that TNFAIP8 might be an important mediator of angiogenesis in DR.

BACKGROUND: Orexin, a neuropeptide derived from the perifornical area of the hypothalamus (PeFLH), promotes the recovery of propofol, isoflurane, and sevoflurane anesthesias, without influencing the induction time. However, whether the orexinergic system also plays a similar role in desflurane anesthesia, which is widely applied in clinical practice owing to its most rapid onset and offset time among all volatile anesthetics, has not yet been studied. In the present study, we explored the effect of the orexinergic system on the consciousness state induced by desflurane anesthesia. METHODS: The c-Fos staining was used to observe the activity changes of orexinergic neurons in the PeFLH and their efferent projection regions under desflurane anesthesia. Chemogenetic and optogenetic techniques were applied to compare the effect of PeFLH orexinergic neurons on the induction, emergence, and maintenance states between desflurane and isoflurane anesthesias. Orexinergic terminals in the paraventricular thalamic nucleus (PVT) were manipulated with pharmacologic, chemogenetic, and optogenetic techniques to assess the effect of orexinergic circuitry on desflurane anesthesia. RESULTS: Desflurane anesthesia inhibited the activity of orexinergic neurons in the PeFLH, as well as the neuronal activity in PVT, basal forebrain, dorsal raphe nucleus, and ventral tegmental area, as demonstrated by c-Fos staining. Activation of PeFLH orexinergic neurons prolonged the induction time and accelerated emergence from desflurane anesthesia but only influenced the emergence in isoflurane anesthesia, as demonstrated by chemogenetic and pharmacologic techniques. Meanwhile, optical activation of orexinergic neurons exhibited a long-lasting inhibitory effect on burst-suppression ratio (BSR) under desflurane anesthesia, and the effect may be contributed by the orexinergic PeFLH-PVT circuitry. The orexin-2 receptor (OX2R), but not orexin-1 receptor (OX1R), in the PVT, which had been inhibited most significantly by desflurane, mediated the proemergence effect of desflurane anesthesia. CONCLUSIONS: We discovered, for the first time, that orexinergic neurons in the PeFLH could not only influence the maintenance and emergence from isoflurane and desflurane anesthesias but also affect the induction under desflurane anesthesia. Furthermore, this specific effect is probably mediated by orexinergic PeFLH-PVT circuitry, especially OX2Rs in the PVT.

Objective:To evaluate the effect of hydrogen on lung injury induced by extremity ischemia-reperfusion (I/R) in elderly patients.Methods:Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ elderly patients, aged 65-75 yr, with height 155-180 cm, weighing 50-75 kg, undergoing lower limb surgery under spinal anesthesia, were divided into 2 groups ( n=30 each) using a random number table method: hydrogen inhalation group (H group) and control group (C group). In H group, 67% hydrogen-33% oxygen was inhaled through the nasal catheter until the end of surgery starting from the completion of anesthesia.In group C, 33% oxygen was inhaled through the nasal catheter until the end of surgery after the completion of anesthesia.Blood samples from the radial artery were collected before anesthesia and at 60 min after tourniquet deflation.Blood gas analysis was performed to determine and record arterial oxygen partial pressure (PaO 2) and arterial carbon dioxide partial pressure (PaCO 2), and alveolar-arterial partial pressure of oxygen difference (A-aDO 2), oxygenation index (OI) and respiratory index (RI) were calculated.Pulmonary surfactant protein D (SP-D) and interleukin-6 (IL-6) concentrations in serum were measured by enzyme-linked immuno sorbent assay.ICU stay time and incidence of pulmonary complications within 7 days after operation were recorded. Results:Compared with group C, PaO 2 and OI were significantly increased, RI and A-aDO 2 were decreased, SP-D and IL-6 concentrations in serum were decreased at 60 min after tourniquet deflation, and ICU stay time was shortened ( P<0.05), and no significant change was found in the incidence of pulmonary complications within 7 days after surgery in group H ( P>0.05). Conclusion:Hydrogen can reduce the lung injury induced by extremity I/R, and the mechanism may be related to the reduction of inflammatory response in elderly patients.

With the development of technology, adjuvant immunotherapy has become a promising strategy for prevention of postoperative tumor regression and metastasis by stimulating the host immune response. However, the therapeutic effects are still unsatisfactory due to the lack of synergy between different methods. In this study, an efficient synergistic immunotherapy system based on injectable sodium alginate hydrogels was designed to inhibit in situ recurrence and metastasis at the same time. On the one hand, an injectable sodium alginate (SA) hydrogel microsystem loaded with toll-like receptor (TLR) agonists (CpG ODNs) was synthesized for inhibiting in situ recurrence, and then carcinoembryonic antigen (CEA) probe was also added to detect CEA based on fluorescence resonance energy transfer (FRET) technology to monitor the occurrence and development of tumor recurrence. On the other hand, an anti-programmed cell death 1 ligand 1 antibody (anti-PD-L1)-modified SA nanogel loaded with indocyanine green (ICG@SA-anti-PD-L1 nanogel) was prepared for diagnosing and inhibiting lung metastasis by assisting orthotopic tumor therapy. In vitro and in vivo results demonstrated that this SA micro/nanosystem could monitor and inhibit postoperative recurrence and metastasis. We hope that this micro/nano-synergistic system will become an effective strategy for postoperative adjuvant immunotherapy.