RALF22 promotes plant immunity and amplifies the Pep3 immune signal.

Rapid alkalinization factors (RALFs) in plants have been reported to dampen pathogen-associated molecular pattern (PAMP)-triggered immunity via suppressing PAMP-induced complex formation between the pattern recognition receptor (PRR) and its co-receptor BAK1. However, the direct and positive role of RALFs in plant immunity remains largely unknown. Herein, we report the direct and positive roles of a typical RALF, RALF22, in plant immunity. RALF22 alone directly elicited a variety of typical immune responses and triggered resistance against the devastating necrotrophic fungal pathogen Sclerotinia sclerotiorum in a FERONIA (FER)-dependent manner. LORELEI (LRE)-like glycosylphosphatidylinositol (GPI)-anchored protein 1 (LLG1) and NADPH oxidase RBOHD were required for RALF22-elicited reactive oxygen species (ROS) generation. The mutation of cysteines conserved in the C terminus of RALFs abolished, while the constitutive formation of two disulfide bridges between these cysteines promoted the RALF22-elicited ROS production and resistance against S. sclerotiorum, demonstrating the requirement of these cysteines in the functions of RALF22 in plant immunity. Furthermore, RALF22 amplified the Pep3-induced immune signal by dramatically increasing the abundance of PROPEP3 transcript and protein. Supply with RALF22 induced resistance against S. sclerotiorum in Brassica crop plants. Collectively, our results reveal that RALF22 triggers immune responses and augments the Pep3-induced immune signal in a FER-dependent manner, and exhibits the potential to be exploited as an immune elicitor in crop protection.

Exosomes from ADSCs ameliorate nerve damage in the hippocampus caused by post traumatic brain injury via the delivery of circ-Scmh1 promoting microglial M2 polarization.

BACKGROUND: Traumatic brain injury (TBI) is an urgent global health issue. Neuroinflammation, due partially to microglia, can worsen or even cause neuropsychiatric disorders after a TBI. An increasing number of studies have found that adipose-derived stem cell (ADSC) derived exosomes can alleviate many diseases by delivering non-coding RNAs including circRNA and miRNAs, but the mechanism of action remains unclear. METHODS: In the present investigation, we produced a TBI mouse model and isolated exosomes from their ADSCs before and after an hypoxic pretreatment. We then used next generation sequencing (NGS) to identify differentially expressed circRNAs and luciferase report assays to determine the relationship between the different noncoding RNAs (miRNA, circRNA and mRNA). RESULTS: The results show that we successfully isolated ADSCs which possessed a multidirectional differentiation potential. We then isolated exosomes from untreated ADSCs (Exos) and from hypoxia pretreated ADSCs (HExos). The HExos significantly decreased hippocampal nerve injury after TBI by decreasing M1 microglia mediated inflammatory cytokine expression and caused recovery of cognitive function. NGS data revealed that abnormal circ-Scmh1 expression plays a role in HExo mediated brain tissue preservation after TBI. Furthermore, luciferase report analysis found that miR-154-5p and STAT6 were the targets for circ-Scmh1. Interestingly, miR-154-5p overexpression or STAT6 inhibition reversed the circ-Scmh1 induced M2 microglial polarization. Overexpression of circ-Scmh1 increased the therapeutic effect of Exo on hippocampal nerve injury after TBI by promotion of M2 microglial polarization and decreased inflammatory induced hippocampal nerve injury. CONCLUSION: Taken together, we found that exosomes from ADSCs ameliorate nerve damage in the hippocampus post TBI through the delivery of circ-Scmh1 and the promotion of microglial M2 polarization.

Tomato LysM receptor kinase 4 mediates chitin-elicited fungal resistance in both leaves and fruit.

Fungal infection is a major cause of crop and fruit losses. Recognition of chitin, a component of fungal cell walls, endows plants with enhanced fungal resistance. Here, we found that mutation of tomato LysM receptor kinase 4 (SlLYK4) and chitin elicitor receptor kinase 1 (SlCERK1) impaired chitin-induced immune responses in tomato leaves. Compared with the wild type, sllyk4 and slcerk1 mutant leaves were more susceptible to Botrytis cinerea (gray mold). SlLYK4 extracellular domain showed strong binding affinity to chitin, and the binding of SlLYK4 induced SlLYK4-SlCERK1 association. Remarkably, qRT-PCR analysis indicated that SlLYK4 was highly expressed in tomato fruit, and ß-GLUCURONIDASE (GUS) expression driven by the SlLYK4 promoter was observed in tomato fruit. Furthermore, SlLYK4 overexpression enhanced disease resistance not only in leaves but also in fruit. Our study suggests that chitin-mediated immunity plays a role in fruit, providing a possible way to reduce fungal infection-related fruit losses by enhancing the chitin-induced immune responses.

A therapeutic regimen of ceftazidime-avibactam for a critical patient receiving prolonged intermittent renal replacement therapy.

The present report firstly described a critically ill patient receiving a dosing regimen of ceftazidime-avibactam (CAZ-AVI) (1.875g q24h) to eliminate multidrug-resistant Klebsiella pneumoniae and a scheduled time for prolonged intermittent renal replacement therapy (PIRRT) every 48h (6h-session beginning 12h after the previous dosage on hemodialysis day). This dosing regimen for CAZ-AVI and a scheduled time for PIRRT allowed pharmacodynamic parameters of ceftazidime and avibactam to have little difference on hemodialysis and non-hemodialysis days so that we can maintain a relatively stable drug concentration. Our report highlighted not only the importance of dosing regimens in patients with PIRRT but also the significance of hemodialysis time points during the dosing interval. The innovative therapeutic plan proved to be suitable for patients infected with Klebsiella pneumoniae when on PIRRT according to the trough plasma concentrations of ceftazidime and avibactam which were maintained above the minimum inhibitory concentration during the dosing interval.

Neuroprotection of NSC Therapy is Superior to Glibenclamide in Cardiac Arrest-Induced Brain Injury via Neuroinflammation Regulation.

Cardiac arrest (CA) is common and devastating, and neuroprotective therapies for brain injury after CA remain limited. Neuroinflammation has been a target for two promising but underdeveloped post-CA therapies: neural stem cell (NSC) engrafting and glibenclamide (GBC). It is critical to understand whether one therapy has superior efficacy over the other and to further understand their immunomodulatory mechanisms. In this study, we aimed to evaluate and compare the therapeutic effects of NSC and GBC therapies post-CA. In in vitro studies, BV2 cells underwent oxygen-glucose deprivation (OGD) for three hours and were then treated with GBC or co-cultured with human NSCs (hNSCs). Microglial polarization phenotype and TLR4/NLRP3 inflammatory pathway proteins were detected by immunofluorescence staining. Twenty-four Wistar rats were randomly assigned to three groups (control, GBC, and hNSCs, N = 8/group). After 8 min of asphyxial CA, GBC was injected intraperitoneally or hNSCs were administered intranasally in the treatment groups. Neurological-deficit scores (NDSs) were assessed at 24, 48, and 72 h after return of spontaneous circulation (ROSC). Immunofluorescence was used to track hNSCs and quantitatively evaluate microglial activation subtype and polarization. The expression of TLR4/NLRP3 pathway-related proteins was quantified via Western blot. The in vitro studies showed the highest proportion of activated BV2 cells with an increased expression of TLR4/NLRP3 signaling proteins were found in the OGD group compared to OGD + GBC and OGD + hNSCs groups. NDS showed significant improvement after CA in hNSC and GBC groups compared to controls, and hNSC treatment was superior to GBC treatment. The hNSC group had more inactive morphology and anti-inflammatory phenotype of microglia. The quantified expression of TLR4/NLRP3 pathway-related proteins was significantly suppressed by both treatments, and the suppression was more significant in the hNSC group compared to the GBC group. hNSC and GBC therapy regulate microglial activation and the neuroinflammatory response in the brain after CA through TLR4/NLRP3 signaling and exert multiple neuroprotective effects, including improved neurological function and shortened time of severe neurological deficit. In addition, hNSCs displayed superior inflammatory regulation over GBC.

Effect of Graded Targeted Temperature Management on Cerebral Glucose Spatiotemporal Characteristics after Cardiac Arrest.

Cardiac arrest (CA) is a fatal disease with high rates of neurological impairment. At present, targeted temperature management (TTM) is the only strategy with firm clinical evidence to prove its effectiveness. However, there is still controversy on the implementation of TTM, particularly on its depth, with a lack of elucidated underlying therapeutic mechanisms. Six Wistar rats were subjected to 8 min asphyxia-CA and randomly divided into TTM at 33oC(n=3) or 35° C groups (n=3). The spatiotemporal characteristics of cerebral glucose metabolism after CA were investigated by 18F-FDG microPET/CT. Myelin Basic Protein (MBP) immunofluorescence staining was used to assess acute injury and recovery of oligodendrocytes. Functional recovery was evaluated using the neurological deficit score (NDS). There was a significant improvement in functional recovery by NDS (p < 0.05) in the 33oC group compared with the 35° C group. Glucose metabolism of the 33° C group was higher than that of the 35oC group early after resuscitation (within 10 minutes). Immunofluorescence analysis showed that positive MBP signals in the cortex and hippocampus in the 33oC group were greater than in the 35oC group. In conclusion, compared to 35oC TTM, 33° C TTM changed the spatiotemporal characteristics of brain glucose metabolisms with improved neurological function, which may be through oligodendrocyte participation.

The copper-associated protein STEAP2 correlated with glioma prognosis and immune infiltration.

High-grade glioma is characterized by cell heterogeneity, gene mutations, and poor prognosis. Abnormal copper homeostasis affects the pathogenesis of glioma, but the underlying mechanisms and involved proteins are unknown. Here, we selected 90 copper-related proteins and verified their expression differences in glioma and normal tissues in the TCGA cohort followed by GO and KEGG clustering analyses. We then developed and validated a prognostic model. Moreover, we examined the mutation burden of copper-related proteins and discussed the differences in the immune microenvironment in the high- and low-risk groups. Furthermore, we focused on STEAP2 and demonstrated that STEAP2 expression was relatively low in tumor tissues compared to normal tissues, implying a favorable prognosis. Our findings provide a foundation for future research targeting copper-related proteins and their immune microenvironment to improve prognosis and responses to immunotherapy.

OsASR6 Alleviates Rice Resistance to Xanthomonas oryzae via Transcriptional Suppression of OsCIPK15.

The plant-specific ASR (abscisic acid, stress and ripening) transcription factors are pivotal regulators of plant responses to abiotic stresses. However, their functions in plant disease resistance remain largely unknown. In this study, we revealed the role of OsASR6 in rice plants' resistance to two important bacterial diseases caused by Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc) and elucidated the mechanisms underlying OsASR6-regulated resistance. The expression of OsASR6 was strongly elevated in response to both Xoo and Xoc challenges. Silencing of OsASR6 in OsASR6-RNAi transgenic plants markedly enhanced rice resistance to the two bacterial pathogens. Moreover, comparative transcriptome analyses for OsASR6-RNAi and wild-type plants inoculated and uninoculated with Xoc demonstrated that OsASR6 suppressed rice resistance to Xoc by comprehensively fine-tuning CIPK15- and WRKY45-1-mediated immunity, SA signaling and redox homeostasis. Further luciferase reporter assays confirmed that OsASR6 negatively regulated CIPK15 but not WRKY45-1 expression in planta. Overexpression of OsCIPK15 strongly enhanced rice resistance to Xoo and Xoc. Collectively, these results reveal that OsASR6 alleviates rice resistance through the transcriptional suppression of OsCIPK15, and thus links calcium signaling to rice resistance against X. oryzae. Our findings provide insight into the mechanisms underlying OsASR6-mediated regulation of rice resistance to X. oryzae.

Genome-Wide Identification of Rapid Alkalinization Factor Family in Brassica napus and Functional Analysis of BnRALF10 in Immunity to Sclerotinia sclerotiorum.

Rapid alkalinization factors (RALFs) were recently reported to be important players in plant immunity. Nevertheless, the signaling underlying RALF-triggered immunity in crop species against necrotrophic pathogens remains largely unknown. In this study, RALF family in the important oil crop oilseed rape (Brassica napus) was identified and functions of BnRALF10 in immunity against the devastating necrotrophic pathogen Sclerotinia sclerotiorum as well as the signaling underlying this immunity were revealed. The oilseed rape genome carried 61 RALFs, half of them were atypical, containing a less conserved YISY motif and lacking a RRXL motif or a pair of cysteines. Family-wide gene expression analyses demonstrated that patterns of expression in response to S. sclerotiorum infection and DAMP and PAMP treatments were generally RALF- and stimulus-specific. Most significantly responsive BnRALF genes were expressionally up-regulated by S. sclerotiorum, while in contrast, more BnRALF genes were down-regulated by BnPep5 and SsNLP1. These results indicate that members of BnRALF family are likely differentially involved in plant immunity. Functional analyses revealed that BnRALF10 provoked diverse immune responses in oilseed rape and stimulated resistance to S. sclerotiorum. These data support BnRALF10 to function as a DAMP to play a positive role in plant immunity. BnRALF10 interacted with BnFER. Silencing of BnFER decreased BnRALF10-induced reactive oxygen species (ROS) production and compromised rape resistance to S. sclerotiorum. These results back BnFER to be a receptor of BnRALF10. Furthermore, quantitative proteomic analysis identified dozens of BnRALF10-elicited defense (RED) proteins, which respond to BnRALF10 in protein abundance and play a role in defense. Our results revealed that BnRALF10 modulated the abundance of RED proteins to fine tune plant immunity. Collectively, our results provided some insights into the functions of oilseed rape RALFs and the signaling underlying BnRALF-triggered immunity.

Structural and Functional Alterations of Substantia Nigra and Associations With Anxiety and Depressive Symptoms Following Traumatic Brain Injury.

Backgrounds: Although there are a certain number of studies dedicated to the disturbances of the dopaminergic system induced by traumatic brain injury (TBI), the associations of abnormal dopaminergic systems with post-traumatic anxiety and depressive disorders and their underlying mechanisms have not been clarified yet. In the midbrain, dopaminergic neurons are mainly situated in the substantia nigra (SN) and the ventral tegmental area (VTA). Thus, we selected SN and VTA as regions of interest and performed a seed-based global correlation to evaluate the altered functional connectivity throughout the dopaminergic system post-TBI. Methods: Thirty-three individuals with TBI and 21 healthy controls were recruited in the study. Anxiety and depressive symptoms were examined by the Hospital Anxiety and Depression Scale. All MRI data were collected using a Siemens Prisma 3.0 Tesla MRI system. The volume of SN and the global functional connectivity of the SN and VTA were analyzed. Results: In the present study, patients with TBI reported more anxiety and depressive symptoms. More importantly, some structural and functional alterations, such as smaller SN and reduced functional connectivity in the left SN, were seen in individuals with TBI. Patients with TBI had smaller substantia nigra on both right and left sides, and the left substantia nigra was relatively small in contrast with the right one. Among these findings, functional connectivity between left SN and left angular gyrus was positively associated with post-traumatic anxiety symptoms and negatively associated with depressive symptoms. Conclusions: The TBI causes leftward lateralization of structural and functional alterations in the substantia nigra. An impaired mesocortical functional connectivity might be implicated in post-traumatic anxiety and depression.

Decreased Interhemispheric Functional Connectivity and Its Associations with Clinical Correlates following Traumatic Brain Injury.

Objective: To explore the interhemispheric functional coordination following traumatic brain injury (TBI) and its association with posttraumatic anxiety and depressive symptoms. Methods: This was a combination of a retrospective cohort study and a cross-sectional observational study. We investigated the functional coordination between hemispheres by voxel-mirrored homotopic connectivity (VMHC). Grey matter volumes were examined by voxel-based morphometry (VBM), and microstructural integrity of the corpus callosum (CC) was assessed by diffusion tension imaging (DTI). The anxiety and depressive symptoms were evaluated with the Hospital Anxiety and Depression Scale. Results: The VMHC values of the bilateral middle temporal gyrus (MTG) and orbital middle frontal gyrus (MFG) were significantly decreased in TBI patients versus the healthy controls. Weakened homotopic functional connectivity (FC) in the bilateral orbital MFG is moderate positively correlated with anxiety and depressive symptoms. The white matter integrity in the CC was extensively reduced in TBI patients. In the receiver operating characteristic analysis, the VMHC value of the orbital MFG could distinguish TBI from HC with an area under the curve of 0.939 (sensitivity of 1 and specificity of 0.867). Conclusion: TBI disrupts the interhemispheric functional and structural connection, which is correlated with posttraumatic mood disorders. These findings may serve as a clinical indicator for diagnosis.

Classification and Characteristics of Mesenchymal Stem Cells and Its Potential Therapeutic Mechanisms and Applications against Ischemic Stroke.

Ischemic stroke is a serious cerebral disease that often induces death and long-term disability. As a currently available therapy for recanalization after ischemic stroke, thrombolysis, including intravenous thrombolysis and endovascular therapy, still cannot be applicable to all patients due to the narrow time window. Mesenchymal stem cell (MSC) transplantation therapy, which can trigger neuronal regeneration and repair, has been considered as a significant advance in treatment of ischemic stroke. MSC transplantation therapy has exhibited its potential to improve the neurological function in ischemic stroke. Our review describes the current progress and future perspective of MSC transplantation therapy in ischemic stroke treatment, including cell types, transplantation approaches, therapeutic mechanisms, and preliminary clinical trials of MSC transplantation, for providing us an update role of MSC transplantation in ischemic stroke treatment.

Carbon Dioxide Tornado-Type Atmospheric-Pressure-Plasma-Jet-Processed rGO-SnO2 Nanocomposites for Symmetric Supercapacitors.

Pastes containing reduced graphene oxide (rGO) and SnCl2 solution were screen printed on carbon cloth and then calcined using a CO2 tornado-type atmospheric-pressure plasma jet (APPJ). The tornado circulation of the plasma gas enhances the mixing of the reactive plasma species and thus ensures better reaction uniformity. Scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS) were performed to characterize the synthesized rGO-SnO2 nanocomposites on carbon cloth. After CO2 tornado-type APPJ treatment, the pastes were converted into rGO-SnO2 nanocomposites for use as the active electrode materials of polyvinyl alcohol (PVA)-H2SO4 gel-electrolyte flexible supercapacitors (SCs). Various APPJ scanning times were tested to obtain SCs with optimized performance. With seven APPJ scans, the SC achieved the best areal capacitance of 37.17 mF/cm2 in Galvanostatic charging/discharging (GCD) and a capacitance retention rate of 84.2% after 10,000-cycle cyclic voltammetry (CV) tests. The capacitance contribution ratio, calculated as pseudocapacitance/electrical double layer capacitance (PC/EDLC), is ~50/50 as analyzed by the Trasatti method. GCD data were also analyzed to obtain Ragone plots; these indicated an energy density comparable to those of SCs processed using a fixed-point nitrogen APPJ in our previous study.

Association of Anxiety and Depressive Symptoms with Memory Function following Traumatic Brain Injury.

INTRODUCTION: Memory impairment and mood disorders are among the most troubling sequelae following traumatic brain injury (TBI). The relationships between comorbid psychiatric disorders and memory function have not been well illustrated. The aim of the study was to explore the relationships of comorbid anxiety and depressive symptoms with memory function following TBI. METHODS: A total of 46 TBI participants across all levels of injury and 23 healthy controls were enrolled in this case-control study. Wechsler Memory Scale-Chinese Revision (WMS-CR) picture, recognition, associative learning, comprehension memory, and digit span were administered to evaluate several categories of memory capacity. The Hospital Anxiety and Depression Scale (HADS) was employed to evaluate the anxiety and depressive symptoms. Stepwise multiple linear regressions were conducted. RESULTS: Compared to healthy controls, the participants with TBI reported more anxiety and depressive symptoms. In the meanwhile, they performed more poorly on memory tests, showing 1.84 SDs, 1.07 SDs, and 0.68 SDs below healthy participants on visuospatial memory, working memory, and verbal memory, respectively. A variety of variables, including HADS depression, HADS anxiety, age, GCS, and education were associated with posttraumatic memory function in the bivariate models. The stepwise multiple linear regressions demonstrated a negative association between HADS depression and posttraumatic memory function, especially performance on visuospatial and verbal memory and a positive association between education and posttraumatic memory function. CONCLUSION: More depressive symptoms rather than anxiety symptoms and less years of education are significant predictors for posttraumatic memory dysfunction.

Targeted temperature management and early neuro-prognostication after cardiac arrest.

Targeted temperature management (TTM) is a recommended neuroprotective intervention for coma after out-of-hospital cardiac arrest (OHCA). However, controversies exist concerning the proper implementation and overall efficacy of post-CA TTM, particularly related to optimal timing and depth of TTM and cooling methods. A review of the literature finds that optimizing and individualizing TTM remains an open question requiring further clinical investigation. This paper will summarize the preclinical and clinical trial data to-date, current recommendations, and future directions of this therapy, including new cooling methods under investigation. For now, early induction, maintenance for at least 24 hours, and slow rewarming utilizing endovascular methods may be preferred. Moreover, timely and accurate neuro-prognostication is valuable for guiding ethical and cost-effective management of post-CA coma. Current evidence for early neuro-prognostication after TTM suggests that a combination of initial prediction models, biomarkers, neuroimaging, and electrophysiological methods is the optimal strategy in predicting neurological functional outcomes.

Comprehensive epigenetic analysis of m6A modification in the hippocampal injury of diabetic rats.

Aim: To study RNA N6-methyladenosine (m6A) modification in the diabetic hippocampus. Methods: Behavioral tests and staining were performed to evaluate the damage to the diabetic hippocampus in model rats. Western blotting was performed to investigate the expression of methylation-related enzymes, and flow cytometry was used to demonstrate HT22 cell apoptosis. M6A and RNA sequencing analyses were conducted to profile m6A-tagged transcripts in the diabetic hippocampus. Results: The rat models of diabetes mellitus suffered from cognitive disorders and hippocampal neuron damage. High glucose levels altered the expression of methylation-related enzymes. A total of 4890 differentially methylated m6A peaks and 63 differentially expressed genes and differentially methylated m6A sites were identified. Conclusion: The findings suggest that m6A modification is altered in the diabetic hippocampus and provide new insight into diabetic hippocampal injury.

Endoscopic surgical treatment of Cushing's disease: A single-center experience of cauterization of peritumoral tissues.

Endoscopic transsphenoidal surgery is a form of treatment for Cushing's disease that is initially preferred compared with other types of treatment. Peritumoral tissue is inactivated by cauterization if a clear border of the lesion cannot be identified. In order to compare the surgical outcomes and post-operative complications between patients who underwent inactivation (cauterization of peritumoral tissues is referred to as inactivation) and those who did not, the medical records of patients treated between January 2010 and June 2016 were retrospectively reviewed. Furthermore, the results of conventional examinations performed in order to diagnose and locate the tumors, including neuroimaging, high-dose dexamethasone suppression tests and/or bilateral inferior petrosal sinus sampling, were collected. A total of 79 consecutive patients with Cushing's disease were included in the present analysis. Inactivation of peritumoral tissue had been applied in 35 (44.3%) of the cases. A gross total resection was achieved in 73 (92.4%) of the cases, while partial resection had been performed in the remaining six (7.6%). Early post-operative endocrinological remission was attained in 71 (89.8%) of the patients. A total of seven cases suffered from hypopituitarism, while cerebrospinal fluid rhinorrhea and brain stem infarction were recorded in four cases and one case, respectively. During the follow-up, the duration of which ranged between 11 and 62 months, recurrence was documented in five patients, as determined by endocrinal examinations. All of the cases that were initially treated unsuccessfully and those with recurrence achieved endocrinological remission following radiotherapy. Enhanced pituitary magnetic resonance imaging revealed no further enlargement of tumors during follow-up. The extent of endocrinological remission, recurrence and post-operative complications did not significantly differ between patients who underwent inactivation and those who did not. In conclusion, inactivation of peritumoral tissue by cauterization achieved satisfactory results in patients with intricate lesions, however it is comparable to conventional procedures.

A Standard Operation Procedure of Clean and Fast Craniotomy Technique for Retrosigmoid Approach.

Clean and fast craniotomy and closure (CAC) for retrosigmoid approach with safe and satisfactory exposure remains our primary goal. A standard operation procedure (SOP) of retrosigmoid approach was developed and reviewed. Between January 2015 and January 2017, 97 patients suffering various lesions underwent surgeries using this technique in the Department of Neurosurgery, Qilu Hospital of Shandong University. The records concerning time of craniotomies, blood losses, and complications were reviewed. By applying this SOP, a craniotomy generally took 15 to 35 minutes, with an average of around 25 minutes. Six cases had a blood loss of more than 30 mL during craniotomy. One patient had cerebrospinal fluid rhinorrhea and another case had subcutaneous effusion. There was no record of venous sinus injury or wound infection. To sum up, the SOP of retrosigmoid approach is simple, reliable and bloodless. In addition to avoiding complications such as venous sinus injury effectively, the SOP also benefits training of residents and early recovery after surgery (ERAS).