Purpose Our study intended to explore how low-dose anti-angiogenic drugs affected anti-tumor immunity of tumor-infiltrating exhausted CD8(+)T cells and achieved better clinical response when combined with immunotherapy. We set out to find potential targets or predictive biomarker on CD8(+)T cells for immunotherapy. Methods We tested different doses of anti-VEGFR2 antibody combined with anti-PD1 antibody to treat LUAD in vivo and analyzed tumor-infiltrating CD8(+)T cells by flow cytometry. CD8(+)T cells overexpressing LAYN were co-cultured with LA795 cell lines to identify the function of LAYN in CD8(+)T cells. We also analyzed clinical samples from advanced LUAD patients treated with anti-angiogenesis therapy combined with immunotherapy. Results Low-dose anti-VEGFR2 antibody combined with anti-PD1 antibody treatment delayed tumor growth and prolonged the survival time of tumor-bearing mice. The number of tumor-infiltrating CD8(+)T cells was reduced and the expression of LAYN was down-regulated in tumor-infiltrating CD8(+)T cells in the low-dose anti-VEGFR2 combination group. It was found that LAYN inhibited the killing function of CD8(+)T cells. In patients with advanced LUAD who received anti-angiogenesis therapy combined with immunotherapy, the LAYN(+)CD8(+)T cell subpopulation in good responders was significantly higher than that in poor responders. Furthermore, we demonstrated the expression of LAYN was regulated by upstream transcription factor NR4A1. Conclusion Low-dose anti-VEGFR2 antibody combined with anti-PD1 antibody therapy promoted anti-tumor immunity and the downregulation of LAYN in tumor-infiltrating CD8(+)T cells played an important role in this process. These findings had implications for improving the efficacy of immune checkpoint blockade therapy and further optimized clinical treatment guidelines in advanced LUAD.

How to develop near-infrared second window (NIR-II, 1000-1700 nm) fluorescent nanoprobes with a uniform size, strong fluorescence signal and good biosafety owns great clinical requirement. Herein we reported that a two photon fluorescent nanoprobe was developed via encapsulating NIR-II-fluorescent molecules into DSPE-PEG, which was effectively endocytosized by cancer cells, and achieved strong NIR-II fluorescence imaging in cancer cells and cancer cell-beard mice models. Prepared NIR-II-fluorescent nanoprobe exhibited rapid metabolism and excellent biocompatibility. In conclusion, the prepared two photon nanoprobe owns good biosafety, and clinical translational prospect in NIR-II fluorescent imaging of tumour in vivo in near future.

[This retracts the article DOI: 10.1016/j.omtn.2017.12.009.].

Recent studies have shown that exposure to sevoflurane in developing brains causes neuronal apoptosis and cognitive dysfunction. “Necroptosis” is a novel pathway of necrosis. We introduced the caspase-specific inhibitor Z-VAD in addition to the receptor-interacting protein kinase 1 (RIPK1) inhibitor Nec-1, to ascertain the existence and importance of necroptosis. Sprague–Dawley rat pups postnatal day 7 were randomly assigned into one of five groups: control, sevoflurane + Z-VAD, sevoflurane + Nec-1, sevoflurane + Z-VAD + Nec-1 and 3% sevoflurane group. Neuronal apoptosis was evaluated by hematoxylin and eosin staining. The MTT assay was performed to evaluate cell viability. Immunofluorescence was employed to measure expression of RIPK1 and RIPK3. Western blots showing expression of RIPK1, RIPK3 and phosphorylation of mixed lineage kinase domain-like (p-MLKL) were used to explore the role of necroptosis. Binding of RIPK1/RIPK3 was detected via co-immunoprecipitation. Finally, the Morris water maze test was used to determine cognitive function. Exposure to 3% sevoflurane for 6 h induced neurotoxicity and inhibited cell viability. Neuron viability was low in the SEV, SEV + Z-VAD and SEV + Nec-1 groups. The study revealed that RIPK1 and RIPK3 protein expression increased significantly, but there was no significant differences between the SEV and SEV + Z-VAD groups. The expression of p-MLKL significantly increased in the SEV and SEV + Z-VAD groups, but not in the SEV + Nec-1 group or SEV + Z-VAD + Nec-1 group compared to the control group. Co-immunoprecipitation results showed that sevoflurane exposure enhanced binding of RIPK1/RIPK3 protein significantly. Blockade of apoptosis and necroptosis alleviated sevoflurane-induced cognitive impairment. Sevoflurane exposure elicited neurotoxicity within neonatal hippocampal neurons and tissues. Blockade of apoptosis or necroptosis alone did not attenuate sevoflurane-induced neurotoxicity (SIN). RIPK1/RIPK3-mediated necroptosis was involved in SIN in hippocampal neurons. SIN could be attenuated only by inhibiting both apoptosis and necroptosis.

目的 采用前瞻性研究对比分析食道引流型喉罩与可弯曲喉罩用于中耳炎手术患者气道管理的有效性和安全性.方法 选择2019年6至12月间在复旦大学附属眼耳鼻喉科医院接受择期中耳炎手术患者130例,年龄18～65岁,ASA分级Ⅰ或Ⅱ级,采用随机数字表法分为食道引流型喉罩(proseal laryngeal mask airway,PLMA)组和可弯曲喉罩(flexible laryngeal mask airway,FLMA)组,每组65例.全麻诱导后,根据患者体重选择合适型号的喉罩,采用经典手指引导法置入喉罩,行机械通气.记录喉罩置入的放置时间、手术时间、苏醒时间、首次放置成功率及总体成功率,记录头颈正中位和侧头位漏气压、15cmH2O压力控制下的呼出潮气量,采用纤维支气管镜观察体位变换前后咽部解剖结构显露情况分级;记录复苏期间的并发症如咽痛、声嘶、呛咳、喉痉挛、胃胀气、恶心呕吐、软组织损伤、喉罩罩体有无血迹、返流误吸等.结果 PLMA放置时间显著短于FLMA(P-0.010 2),而两种喉罩的首次放置成功率和总体放置成功率的差异均无统计学意义.FLMA的侧头位潮气量显著低于正头位(P=0.020 7),且显著低于PLMA组侧头位(P=0.013 4).无论正侧位,PLMA的漏气压均显著高于FLMA(正侧位P均＜0.000 1),而FLMA由正头位改为侧头位后漏气压显著下降(P=0.0128).纤支镜定位分级结果显示各组间差异均无统计学意义.两组患者苏醒时间差异无统计学意义.FLMA组患者苏醒后咽痛(P=0.022 6)和胃胀气(P=0.042 2)显著多于PLMA组,其余并发症两组间差异无统计学意义.结论 PLMA与FLMA均可应用于全身麻醉下中耳炎手术的气道管理,两者有效性和安全性相当.PLMA密封效果更好,在变换头位时,潮气量不易受影响,胃胀气发生率较低,但存在软组织损伤的风险.

Excessively activated microglia exhibit increased migration, resulting in tissue damage and chronic inflammation. Src was confirmed to play an important role in regulation of cell motility following lipopolysaccharide (LPS) treatment. SET8 plays an important part in multiple cellular signal pathways. In this study, we speculated that SET8 is involved in LPS-induced microglial migration via regulation of Src expression. Our study showed that LPS promoted cell migration via augmentation of Src expression in BV2 cells. Moreover, LPS treatment decreased SET8 expression and upregulated the expression of the transcription factor ETS proto-oncogene 1 (ETS1). Overexpression of both SET8 and small interfering ETS1 reversed LPS-induced Src expression and cell migration. The effects of short hairpin SET8 (shSET8) and ETS1 overexpression are the same as the effects of LPS treatment. Decrease of Src expression reversed the shSET8-induced and ETS1 overexpression-induced migration of BV2 cells. Furthermore, SET8 was observed to associate with ETS1. Chromatin immunoprecipitation assay indicated H4K20me1, a downstream target of SET8, in addition to ETS1, was enriched at the Src promoter region. Furthermore, shSET8 increased Src promoter activity and also increased the positive effect of ETS1 overexpression on Src promoter activity. This study shows that SET8 associates with ETS1 to regulate Src expression, which is involved in LPS-induced BV2 cell migration.

BACKGROUND:During emergence from anesthesia after partial and total laryngectomy, excessive airway reflex and systemic hypertension may lead to subcutaneous emphysema, hemorrhage or pneumothorax.METHODS:American Society of Anesthesiologist physical status III and IV male adults undergoing elective laryngectomy were recruited and randomly allocated to receive either dexmedetomidine (group D) or midazolam (group M). The primary outcome was incidence and severity of cough. Pulse oximetry results (SpO2), heart rate (HR), systolic blood pressure (SBP), and diastolic blood pressure (DBP) were also recorded. The visual analog scale and the Ramsay sedation scale were recorded at the points of wakefulness and departure from the post-anesthesia care unit (PACU). Rescue analgesia consumption, the time of spontaneous breath recovery, duration of the PACU stay, and the incidence of adverse effects were also recorded.RESULTS:The prevalence of no coughing was significantly higher in group D than in group M at the points of wakefulness and departure. HR, SBP, and DBP were significantly lower in group D compared with group M, and SpO2 was significantly higher in group D than in group M at the moment of laryngectomy. Pain scores were lower in group D than in group M. The Ramsay score at the point of wakefulness was higher in group D than in group M. There was no difference in time to spontaneous breathing recovery, duration of the PACU stay, and incidence of adverse effects.CONCLUSIONS:Compared with midazolam, dexmedetomidine is an effective alternative to attenuate coughing and hemodynamic changes with a low incidence of adverse events during emergence from anesthesia after partial and total laryngectomy.TRIAL REGISTRATION:NCT03918889 , registered at clinicaltrials.gov, date of registration: March 28, 2019.

Objective: The purpose of this study was to investigate whether a brief exposure to sevoflurane can lead to adverse neurodevelopmental outcomes in children undergoing palatoplasty. Materials and methods: Young children (less than 2 years old) receiving palatoplasty with sevoflurane anesthesia were recruited. The Bayley Scales of Infant Development-Second Edition (BSID-II) assessment was used to evaluate neurodevelopmental outcomes before and after surgery. The levels of plasma neuron-specific enolase and S-100 beta were also measured. Results: This study enrolled 101 patients and 100 healthy children for analysis. The sevoflurane exposure time of all patients was 46.2 +/- 6.5 minutes. There were no statistically significant differences noted between preoperative and postoperative BSID-II scores, plasma neuron-specific enolase and S-100 beta values. When compared with normal healthy children, there were also no statistically significant differences between patients and normal healthy children at the age of 3-4 years in the BSID-II scores (P>0.05). Conclusions: The data did not show any significant evidence of an association between a brief exposure to sevoflurane during early childhood and adverse neurodevelopmental outcomes in children undergoing palatoplasty.

Sevoflurane is the most widely used anaesthetic administered by inhalation. Exposure to sevoflurane in neonatal mice can induce learning deficits and abnormal social behaviours. MicroRNA (miR)-27a-3p, a short, non-coding RNA that functions as a tumour suppressor, is up-regulated after inhalation of anaesthetic, and peroxisome proliferator-activated receptor γ (PPAR-γ) is one of its target genes. The objective of this study was to investigate how the miR-27a-3p-PPAR-γ interaction affects sevoflurane-induced neurotoxicity. A luciferase reporter assay was employed to identify the interaction between miR-27a-3p and PPAR-γ. Primary hippocampal neuron cultures prepared from embryonic day 0 C57BL/6 mice were treated with miR-27a-3p inhibitor or a PPAR-γ agonist to determine the effect of miR-27a-3p and PPAR-γ on sevoflurane-induced cellular damage. Cellular damage was assessed by a flow cytometry assay to detect apoptotic cells, immunofluorescence to detect reactive oxygen species, western blotting to detect NADPH oxidase 1/4 and ELISA to measure inflammatory cytokine levels. In vivo experiments were performed using a sevoflurane-induced anaesthetic mouse model to analyse the effects of miR-27a-3p on neurotoxicity by measuring the number of apoptotic neurons using the Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) method and learning and memory function by employing the Morris water maze test. Our results revealed that PPAR-γ expression was down-regulated by miR-27a-3p following sevoflurane treatment in hippocampal neurons. Down-regulation of miR-27a-3p expression decreased sevoflurane-induced hippocampal neuron apoptosis by decreasing inflammation and oxidative stress-related protein expression through the up-regulation of PPAR-γ. In vivo tests further confirmed that inhibition of miR-27a-3p expression attenuated sevoflurane-induced neuronal apoptosis and learning and memory impairment. Our findings suggest that down-regulation of miR-27a-3p expression ameliorated sevoflurane-induced neurotoxicity and learning and memory impairment through the PPAR-γ signalling pathway. MicroRNA-27a-3p may, therefore, be a potential therapeutic target for preventing or treating sevoflurane-induced neurotoxicity.

Sevoflurane, an inhalational anesthetic, is extensively used during oral cancer surgery. However, the effect of sevoflurane on head and neck squamous cell carcinoma (HNSCC) remains unclear. The objective of the present study was to investigate the effects of sevoflurane on the proliferation, apoptosis and invasion in HNSCC cell lines and the underlying molecular mechanism. The Cell Counting Kit-8 assay was used to evaluate cell proliferation. Apoptosis was analyzed by flow cytometry. Cell invasion was evaluated using the Transwell invasion assay. The expression levels of Akt, p-Akt (Ser473), hypoxia‑inducible factor-1α (HIF-1α), Fas and Bcl-2 were measured by western blotting. Significant inhibition of cell proliferation and induction of apoptosis were observed in FaDu and CAL-27 cells following sevoflurane treatment. The expression of Akt, p-Akt (Ser473) and Bcl-2 was supressed, while that of Fas was significantly increased, which was partly associated with the activation of the HIF-1α pathway. In addition, the results revealed a statistically significant inhibition of cell invasion in the FaDu cell line following exposure to 2 and 4% sevoflurane for 2, 4, 6 and 8 h. Therefore, the present study demonstrated that sevoflurane decreased the malignant behavior of HNSCC cell lines in vitro, which was associated with activation of the HIF-1α pathway.

Long non coding RNAs (lncRNAs) show an encouraging trend in regulating the proliferation of bone marrow-derived mesenchymal stromal cells (BMSCs). The present study investigated the role of lncRNA low expression in tumor (LET) in BMSCs proliferation. Our result showed that LET was down-regulated in rapidly proliferated BMSCs (P < 0.05). Suppression of LET promoted BMSCs proliferation and over-expression of LET inhibited BMSCs proliferation (P < 0.05). LET negatively regulated the expression of transforming growth factor β1 (TGF-β1) in BMSCs (P < 0.05). Knockdown of TGF-β1 reversed the LET suppression-induced BMSCs proliferation (P < 0.05). Moreover, knockdown of TGF-β1 alleviated the LET suppression-induced activation of Wnt/β-catenin pathway in BMSCs. Therefore, we drew the conclusion that LET suppression promoted BMSCs proliferation by up-regulating the expression of TGF-β1 and activating Wnt/β-catenin pathway.

In recent years, long noncoding RNAs (lncRNAs) have been demonstrated to play important roles in the development of human cancer. We assessed the role of lncRNA ANRIL in non-small-cell lung cancer (NSCLC). Quantitative real-time polymerase chain reaction was employed to detect the expression of ANRIL in NSCLC tissues and paired nontumor tissues. The high expression level of ANRIL was positively correlated with advanced tumor-node-metastasis stage and greater tumor diameter. Furthermore, chromatin immunoprecipitation assays confirmed the physical interaction between c-Myc and ANRIL. ANRIL silencing significantly inhibited NSCLC cell proliferation. Together, we showed that ANRIL is involved in the oncogenesis of NSCLC, and ANRIL may be a potential therapeutic target for patients with NSCLC.

Background: Sevoflurane is an inhaled anesthetic commonly used in the pediatric. Recent animal studies suggest that early exposure to high concentration of sevoflurane for a long duration can induce neuroapoptosis and later cognitive dysfunction. However, the neurodevelopmental impact induced by lower concentration and shorter exposure duration of sevoflurane is unclear. To investigate whether early exposure to 2% concentration of sevoflurane for a short duration (clinically relevant usage of sevoflurane) can also induce neuroapoptosis and later cognitive dysfunction.Methods: Rat pups were subjected to control group, 2% sevoflurane for 3 h and 3% sevoflurane for 6 h. TUNEL assay and apoptotic enzyme cleaved caspase-3 measured by western blot were used for detection of neuronal apoptosis in frontal cortex and CA1 region of hippocampus 24 after sevoflurane treatment. Long-term cognitive function was evaluated by Morris water maze and passive avoidance test as the rats grew up.Results: The apoptotic levels in frontal cortex and CA1 region were significantly increased after rats exposed to 3% sevoflurane for 6 h (P < 0.05), but not 2% sevoflurane for 3 h (P > 0.05). Exposure to both 2% sevoflurane for 3 h and 3% sevoflurane for 6 h could cause long-term cognitive dysfunction and animals exposed to 3% sevoflurane for 6 h exhibited worse neurodevelopmental outcomes (P < 0.05).Conclusion: It was suggested that neuronal apoptosis might not contribute to long-term cognitive dysfunction induced by 2% concentration and short exposure time of sevoflurane. Our findings also suggested that the mechanisms of sevoflurane-induced neurodevelopmental impact might be various, depending on the concentration and exposure duration. (C) 2016 Elsevier Masson SAS. All rights reserved.

Background Certain early-phase clinical trials have suggested that bone marrow-derived stem cell transplantation might improve left ventricular function in patients with non-ischaemic dilated cardiomyopathy (NIDCM), whereas others trials have revealed no benefit from this approach. We sought to evaluate the therapeutic effects of bone marrow-derived stem cell therapy on NIDCM. Methods We searched the PubMed, Embase, and Cochrane Central Register of Controlled Trials (CENTRAL) databases (through February 2016) for randomised controlled clinical trials that reported on bone marrow-derived stem cell transplantation for patients with NIDCM with a follow-up period ≥12 months. The co-primary endpoints were changes in mortality rate and left ventricular ejection fraction (LVEF); the secondary endpoints were changes in the 6-minute-walk test (6MWT) and left ventricular chamber size. Seven trials involving bone marrow-derived stem cell therapy that included 482 patients satisfied the inclusion and exclusion criteria. Results Subjects who received bone marrow-derived stem cell therapy exhibited a significant reduction in mortality rate (19.7% in the cell group vs. 27.1% in the control group; 95% confidence interval (CI) –0.16 to –0.00, I 2 = 52%, p = 0.04). Bone marrow-derived stem cell therapy tended to produce LVEF improvement within 6 months (1.83% increase; 95% CI –0.27 to 3.94, I 2 = 74%, p = 0.09) and significantly improved LVEF after mid-term (6–12 months) follow-up (3.53% increase; 95% CI 0.76 to 6.29, I 2 = 88%, p = 0.01). However, this therapy produced no significant benefit in the 6MWT ( p = 0.18). Finally, the transplantation of increased numbers of stem cells resulted in no observable additional benefit with respect to LVEF. Conclusions Bone marrow-derived stem cell therapy might have improved prognoses and appeared to provide moderate benefits in cardiac systolic function at mid-term follow-up. However, this therapy produced no observed improvement in exercise tolerance.

Anaesthetic agents were confirmed to play a role on the tumor angiogenesis. The effect of sevoflurane on tongue squamous cell carcinoma (TSCC) cell has not been investigated. SCC-4 cells were exposed to sevoflurane after simulating hypoxia. Then, both the mRNA and protein level of hypoxia-inducible factor (HIF)-1α and VEGF were detected. The methylation states of the VEGF promoter region were also assessed to reveal the underlying mechanism. Finally, the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza) was administrated to reveal the relationship of DNA methylation on the regulation of the VEGF level. Results showed that sevoflurane attenuated the hypoxia-induced VEGF level without altering the HIF-1α after exposure for 24 and 72 h. Sevoflurane increased the DNA methylation of the VEGF promoter region. The attenuation effect of sevoflurane on hypoxia-induced VEGF level could be blocked by 5-Aza. We concluded that sevoflurane attenuates hypoxia-induced VEGF level via DNA methylation of the promoter region in TSCC cell.

Animal experiments indicate that repeated exposure to ketamine adversely affects the developing brain. Whether it has the same effect on infants remains unclear. We recruited infants who were scheduled for 1 to 3 outpatient laser surgery treatments of benign facial growths with ketamine anesthesia. Patients were assigned to the Ket1, Ket2, or Ket3 group, according to the number of treatments. The Bayley Scales of Infant Development–Second Edition (BSID-II) was used to assess neurodevelopmental outcomes before the first and after the last therapy. Levels of S-100β were also measured. Bayley Scales of Infant Development–Second Edition scores after the last procedure were lower than those before the first surgery in the Ket3 group ( P < .05). S-100β levels after the last procedure were significantly higher than those before the first surgery in all groups ( P < .05). Our results suggest that 3 or more exposures to anesthetic ketamine have the potential to adversely affect neurodevelopment in infants.

Recent studies have shown that dexmedetomidine exerts an anti-inflammatory effect by reducing serum levels of inflammatory factors, however, the up-stream mechanism is still unknown. The transcription factor NF-κB enters the nucleus and promotes the transcription of its target genes, including those encoding the pro-inflammatory cytokines IL-6 and TNF-α. In this study, we established a rat model that simulates a clinical surgical procedure to investigate the anti-inflammatory effect of perineural administration of dexmedetomidine and the underlying mechanism. Dexmedetomidine reduced the sciatic nerve levels of IL-6 and TNF-α at both the mRNA and protein level. Dexmedetomidine also inhibited the translocation of activated NF-κB to the nucleus and the binding activity of NF-κB. The anti-inflammatory effect is confirmed to be dose-dependent. Finally, pyrrolidine dithiocarbamate also reduced the levels of IL-6 and TNF-α and the activation of NF-κB. In conclusion, dexmedetomidine inhibited the nuclear translocation and binding activity of activated NF-κB, thus reducing inflammatory cytokines.

Dexmedetomidine (DEX), a highly specific α2-adrenergic agonist, which exhibits anaesthetic-sparing, analgesia and sympatholytic properties. DEX modulates gene expression, channel activation, transmitter release, inflammatory processes and apoptotic and necrotic cell death. It has also been demonstrated to have protective effects in a variety of animal models of ischemia/reperfusion (I/R) injury, including the intestine, myocardial, renal, lung, cerebral and liver. The broad spectrum of biological activities associated with DEX continues to expand, and its diverse effects suggest that it may offer a novel therapeutic approach for the treatment of human diseases with I/R involvement.

Background: Recent animal experiments have suggested that ketamine administration during development might induce widespread neurodegeneration and long-term cognitive deficits. The underlying mechanism is not fully understood. Methods: Immature rat hippocampal neurons and newborn rats underwent repeated exposure to ketamine, ketamine+inhibitor of hypoxia-inducible factor (HIF)-1α(YC-1), ketamine+inhibitor of reactive oxygen species(ROS) (L-carnitine) or ketamine+Ca2+ blocker(nimodipine). Apoptosis of the hippocampal neurons was analyzed by TUNEL and flow cytometry. Intracellular ROS were measured using 2',7'-dichlorofluorescein diacetate. The expression of HIF- 1α and apoptosis-related proteins was analyzed by western blot or qPCR. As these rats grew, behavioral tests were performed to evaluate cognitive function. Results: The apoptotic rate in the ketamine group was significantly higher than that in the other groups, and the intracellular ROS levels in the ketamine and ketamine+YC-1 groups were higher than those in the other groups. The expression of HIF- 1α, p53, BNIP3 and cleaved caspase-3 proteins increased, and the ratio of Bcl-2/Bax decreased in the ketamine group. The transcriptional levels of HIF-1α in the ketamine and ketamine+YC-1 groups were higher than those in the other groups. Cognitive deficits were found only in the ketamine group. Conclusion: We suggest that ketamine-induced neurodegeneration in neonatal rats, followed by long-term cognitive deficits, might be mediated via the ROS/HIF-1α pathway.

A novel implementation of in situ protein digestion supported by a graphene oxide-immobilized enzyme reactor (GO-IMER) in the MALDI imaging mass spectrometry (IMS) workflow is reported, which enables the simultaneous diagnostic identity and distribution attributes of the proteome on tissue.