Role of adenosine A2A receptors in the loss of consciousness induced by propofol anesthesia.

The mechanism of propofol-anesthesia-induced loss of consciousness (LOC) remains largely unknown. We speculated that the adenosine A2A receptor serves as a vital molecular target in regulating LOC states under propofol anesthesia. c-Fos staining helped observe the changes in the neuronal activity in the nucleus accumbens (NAc). Initially, the adenosine signals in the NAc were measured under propofol anesthesia using fiber photometry recordings. Then, behavior tests and electrophysiological recordings were used to verify the effect of systemic A2A R agonist or antagonist treatment on propofol anesthesia. Next, the microinjection technique was used to clarify the role of the NAc A2A R under propofol anesthesia. Fiber photometry recordings were applied to assess the effect of A2A R agonist or antagonist systemic treatment on adenosine signal alterations in the NAc during propofol anesthesia. Then, as the GABAergic neurons are the main neurons in the NAc, we further measured the neuronal activity of GABAergic neurons. In our study, propofol anesthesia enhanced the neuronal activity in the NAc, and the adenosine signals were increased in the NAc. SCH58261 reduced the LOC time and sedative depth, while CGS21680 increased those via intraperitoneal injection. Additionally, CGS21680 increased the changes in delta, theta, alpha, beta, and low-gamma oscillations in the NAc. Moreover, microinjection of SCH58261 significantly shortened the LOC time, whereas microinjection of CGS21680 into the NAc significantly prolonged the LOC duration. The results illustrated that after A2A R agonist administration, the level of extracellular adenosine signals in the NAc was decreased and the neuronal activity of GABAergic neurons was enhanced, whereas after A2A R antagonist administration via intraperitoneal injection, the opposite occurred. This study reveals the vital role of the A2A R in propofol-induced LOC and that the A2A R could affect the maintenance of propofol anesthesia.

Akkermansia muciniphila improves heat stress-impaired intestinal barrier function by modulating HSP27 in Caco-2 cells.

OBJECTIVE: Heat stress causes an elevation of intestinal epithelial barrier permeability and leads to multiple organ dysfunction in heatstroke. Akkermansia muciniphila (A. muciniphila) plays a role in maintaining intestinal integrity and improving the inflammatory state. This study aimed to investigate whether A. muciniphila could alleviate heat stress-induced dysfunction of intestinal permeability in Caco-2 monolayers and have the preventive effects on heatstroke. METHODS: Human intestinal epithelial Caco-2 cells were preincubated with live or pasteurized A. muciniphila then exposed to heat stress at 43 °C. Transepithelial electrical resistance (TEER) and the flux of horseradish peroxidase (HRP) across cell monolayers were measured to determine intestinal permeability. The levels of the tight junction proteins Occludin, ZO-1 and HSP27 were analyzed by Western blotting. These proteins were immunostained and localized by fluorescence microscopy. TJ morphology was observed using transmission electron microscopy (TEM). RESULTS: Both live and pasteurized A. muciniphila effectively attenuated the decrease in TEER and impairment of intestinal permeability in HRP flux induced by heat exposure. A. muciniphila significantly elevated the expression of Occludin and ZO-1 by promoting HSP27 phosphorylation. The distortion and redistribution of tight junction proteins and disruption of morphology were also effectively prevented by pretreatment with A. muciniphila. CONCLUSION: This study indicates for the first time that both live and pasteurized A. muciniphila play an important protective role against heat-induced permeability dysfunction and epithelial barrier damage.

A Quickly Organized Response to a COVID-19 Positive Case Comeback After Clearance for over 2 Months in Shenzhen, China.

BACKGROUND: The ongoing coronavirus disease 2019 (COVID-19) pandemic has spread quickly throughout the world. So far, there are no standard measures in terms of quick and effective control of new COVID-19 positive patient comeback after COVID-19 clearance for a certain time. Here, we report a quickly organized response from Luohu Hospital Group to a COVID-19 positive case comeback after clearance for over 2 months in Shenzhen, China. METHODS: After 2 months with no new COVID-19 cases in Shenzhen, on August 14, 2020, a supermarket employee living in the Luohu district of Shenzhen was tested positive in a nearby city. She has no any symptoms and signs, with unremarkable laboratory testing and radiological image, so she was soon diagnosed as asymptomatic COVID-19 positive case. Rapid contact tracing revealed that three of her relatives in Shenzhen were infected with COVID-19 and all of them were diagnosed as asymptomatic COVID-19 positive cases. To ensure residents' safety, Luohu hospital group (LHG) mounted a rapid organized response focusing on four measures: local environment management and residents' health monitoring, guidance for resumption of work, education and psychological counseling, and management of patients with fever in outpatient clinics. RESULTS: The LHG being structured as a people-centered, integrated organization responded to residents' medical and psychological needs rapidly, provided 6-hour results for COVID-19 testing, and recleared the city of COVID-19, as evidenced by the processing of 459,381 community samples within 15 days, with universally negative results beyond the originally identified case and her three close relatives. CONCLUSIONS: A quick and effective response from local organization to a new comeback COVID-19 positive case after clearance for a certain time is necessary in terms of ensuring the physical and psychological health of residents, as well as guarantying normal social work.

The adenosine A2A receptor alleviates postoperative delirium-like behaviors by restoring blood cerebrospinal barrier permeability in rats.

Postoperative delirium (POD) is a common post-operative complication in elderly patients that is associated with increased morbidity and mortality. However, the neuropathogenesis of this complication remains unknown. The blood-cerebrospinal fluid barrier (BCB) and brain-blood barrier (BBB) are composed of tight junctions between cells that form physical barriers, and BBB damage plays an important role in the neuropathogenesis of POD. Nevertheless, the role of BCB in POD remains to be elucidated. Herein, we investigated the effect of adenosine A2A receptor (A2A R), a key regulator of the permeability of barriers, on surgery-induced increased permeability of BCB and POD-like behaviors. Open field, buried food, and Y maze tests were used to evaluate behavioral changes in rats after surgery. Levels of tight junction proteins, adherens junction proteins, A2A R, GTP-RhoA, and ROCK2 in the choroid plexus were assessed by western blotting. The concentrations of NaFI and FITC-dextran in the cerebrospinal fluid (CSF) were detected by fluorescence spectrophotometry. Transmission electron microscopy was applied to observe the ultrastructure of the choroid plexus. Surgery/anesthesia decreased the levels of tight junction (e.g., ZO-1, occludin, and claudin1) proteins, increased concentrations of NaFI and FITC-dextran in CSF, damaged the ultrastructure of choroid plexus, and induced POD-like behaviors in rats. An A2A R antagonist alleviated POD-like behaviors in rats. Furthermore, the A2A R antagonist increased the levels of tight junction proteins and restored the permeability of BCB in rats with POD. Fasudil, a selective Rho-associated protein kinase 2 (ROCK2) inhibitor, ameliorated POD-like behaviors induced by A2A R activation. Moreover, fasudil also abolished the increased levels of GTP-RhoA/ROCK2, decreased levels of tight junction proteins, and increased permeability of BCB caused by A2A R activation. Our findings demonstrate that A2A R might participate in regulating BCB permeability in rats with POD via the RhoA/ROCK2 signaling pathway, which suggests the potential of A2A R as a therapeutic target for POD.

Neuronal mechanisms of adenosine A2A receptors in the loss of consciousness induced by propofol general anesthesia with functional magnetic resonance imaging.

Propofol is the most common intravenous anesthetic agent for induction and maintenance of anesthesia, and has been used clinically for more than 30 years. However, the mechanism by which propofol induces loss of consciousness (LOC) remains largely unknown. The adenosine A2A receptor (A2A R) has been extensively proven to have an effect on physiological sleep. It is, therefore, important to investigate the role of A2A R in the induction of LOC using propofol. In the present study, the administration of the highly selective A2A R agonist (CGS21680) and antagonist (SCH58261) was utilized to investigate the function of A2A R under general anesthesia induced by propofol by means of animal behavior studies, resting-state magnetic resonance imaging and c-Fos immunofluorescence staining approaches. Our results show that CGS21680 significantly prolonged the duration of LOC induced by propofol, increased the c-Fos expression in nucleus accumbens (NAc) and suppressed the functional connectivity of NAc-dorsal raphe nucleus (DR) and NAc-cingulate cortex (CG). However, SCH58261 significantly shortened the duration of LOC induced by propofol, decreased the c-Fos expression in NAc, increased the c-Fos expression in DR, and elevated the functional connectivity of NAc-DR and NAc-CG. Collectively, our findings demonstrate the important roles played by A2A R in the LOC induced by propofol and suggest that the neural circuit between NAc-DR maybe controlled by A2A R in the mechanism of anesthesia induced by propofol.

iRNAD: a computational tool for identifying D modification sites in RNA sequence.

MOTIVATION: Dihydrouridine (D) is a common RNA post-transcriptional modification found in eukaryotes, bacteria and a few archaea. The modification can promote the conformational flexibility of individual nucleotide bases. And its levels are increased in cancerous tissues. Therefore, it is necessary to detect D in RNA for further understanding its functional roles. Since wet-experimental techniques for the aim are time-consuming and laborious, it is urgent to develop computational models to identify D modification sites in RNA. RESULTS: We constructed a predictor, called iRNAD, for identifying D modification sites in RNA sequence. In this predictor, the RNA samples derived from five species were encoded by nucleotide chemical property and nucleotide density. Support vector machine was utilized to perform the classification. The final model could produce the overall accuracy of 96.18% with the area under the receiver operating characteristic curve of 0.9839 in jackknife cross-validation test. Furthermore, we performed a series of validations from several aspects and demonstrated the robustness and reliability of the proposed model. AVAILABILITY AND IMPLEMENTATION: A user-friendly web-server called iRNAD can be freely accessible at http://lin-group.cn/server/iRNAD, which will provide convenience and guide to users for further studying D modification.

Importance of respiratory airway management as well as psychological and rehabilitative treatments to COVID-19 patients.

The clinical therapy for severe 2019 coronavirus disease (i.e., COVID-19) sufferers is relatively challenging. Herein, the processes involving salvage of a critical COVID-19 patient were retrospectively analyzed. The condition of an obese female critical COVID-19 sufferer progressively worsened in the initial period after admission. According to her symptoms and examination reports, endotracheal intubation and mechanical ventilation were timely conducted and meanwhile high-dose sedatives and analgesics were administrated. In the later therapeutic phase, however, sedative and analgesic dosages were gradually reduced, and psychological and rehabilitative therapies were conducted, concomitantly with enhancement of airway care to facilitate sputum expectoration. Eventually, the endotracheal tube was feasibly removed after intubation for 18 days and subsequently replaced with noninvasive ventilation and a high-flow nasal cannula oxygen therapy. Intensive airway care alongside psychological and rehabilitative therapies can shorten the mechanical ventilation time and improve the prognosis of COVID-19 sufferers.

MicroRNA-181b-5p attenuates early postoperative cognitive dysfunction by suppressing hippocampal neuroinflammation in mice.

BACKGROUND: Postoperative cognitive dysfunction (POCD) is a common complication after surgery and its occurrence is associated with increased morbidity and mortality. However, the pathophysiology of this complication remains largely unknown. Efforts to identify causes of POCD have focused on the hippocampal neuroinflammation. Recently, accumulated evidence indicates that NeurimmiRs, a subset of microRNAs (miRNAs), which modulate both neuronal and immune processes, play an important role in neuroinflammation. However, the impact of NeurimmiRs on POCD has not been investigated. We hypothesized that NeurimmiRs is involved in surgery-induced cognitive impairment in adult mice via mediating hippocampal neuroinflammation. METHODS: MicroRNA(miR)-181b-5p was found to be downregulated in the hippocampi of mice with POCD using microRNA array, which was also verified in vivo in the mouse model of POCD by Quantitative real-time polymerase chain reaction (qPCR). Subsequently, the expression of miR-181b-5p was measured in lipopolysaccharide (LPS) stimulated BV-2 microglial cells and hippocampal tissues of the mice with POCD. Then, loss of function and overexpression studies were performed by transfection with miR-181b-5p mimic/ inhibitor in cultured BV-2 cell lines and intrahippocampal injection of miR-181b-5p agomir before Surgery/Anesthesia, to identify the role of miR-181b-5p in neuroinflammation and cognitive impairments. QPCR, western blot and ELISA were used to determine the expression of proinflammatory mediators. Immunofluorescence staining was applied to evaluate the activation of microglia. Furthermore, we used bioinformatics analysis and dual-luciferase assay to predict and verify the potential target of miR-181b-5p. RESULTS: The results indicated that miR-181b-5p mimic could repress the mRNA and protein expression of proinflammatory mediators, including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and monocyte chemoattractant protein (MCP)-1 in LPS-stimulated BV-2 microglial cells, while the miR-181b-5p inhibitor induced upregulation of the above-mentioned proinflammatory factors. Further bioinformatics analysis showed that miR-181b-5p was predicted to potentially target the 3'-untranslated region (UTR) of TNF-α, and binding sites of miR-181b-5p in the 3'-UTR of TNF-α were identified by dual-luciferase assay. Importantly, injecting miR-181b-5p agomir into the hippocampus of mice before surgery, ameliorated the hippocampus-dependent memory, and was accompanied by downregulation of proinflammatory factors expression and reduced microglial activation in the hippocampus of POCD mice. CONCLUSIONS: Collectively, these findings suggest that miR-181b-5p attenuates early POCD by suppressing hippocampal neuroinflammation in mice. They also highlight the importance of studying miRNAs in the context of POCD and identify miR-181b-5p as a novel potential therapeutic target for improving POCD.

MicroRNA-146a protects against cognitive decline induced by surgical trauma by suppressing hippocampal neuroinflammation in mice.

Postoperative cognitive dysfunction (POCD) is a common postoperative complication that is associated with increased morbidity and mortality. However, the neuropathogenesis of this complication remains largely unknown. Neuroinflammation, in particular hippocampal inflammation, contributes to POCD. Recently, increasing evidence has supported the involvement of microRNAs (miRNAs) in the regulation of neuroinflammation in human neurological disorders. In the present study, we investigated the role of miR-146a, a key regulator of the innate immune response, in surgery-induced hippocampal inflammation and cognitive impairment. The expression of miR-146a was measured in BV-2 microglial cells stimulated with lipopolysaccharide (LPS) and hippocampal tissues of mice with POCD. Loss of function and overexpression studies were performed via transfection with miR-146a mimic/inhibitor in cultured BV-2 cell lines and intrahippocampal injection of miR-146a agomir/antagomir before surgery/anesthesia to identify the role of miR-146a in neuroinflammation and cognitive impairment. QPCR, Western blot and ELISA were used to determine the expression levels of downstream adaptor proteins and proinflammatory cytokines. Immunofluorescence staining was applied to evaluate the activation of microglia. Increased expression of miR-146a was observed in BV-2 microglial cells stimulated with LPS and hippocampal tissues of mice with POCD. Modulation of miR-146a expression via transfection of microglia with miR-146a mimic or inhibitor regulated the mRNA and protein expression levels of downstream targets of miR-146a (IRAK1 and TRAF6) as well as the release of proinflammatory cytokines (TNF-α, IL-1ß and IL-6). In addition, overexpression of miR-146a attenuated hippocampus-dependent learning and memory impairment in mice with POCD, which was accompanied by decreased expression of the IRAK1/TRAF6/nuclear factor (NF)-κB pathway and downregulation of microglial activation in the hippocampus. Conversely, knockdown of miR-146a expression may exacerbate hippocampus-dependent learning and memory deficiency and hippocampal inflammation in mice with POCD. Collectively, our findings demonstrate the important role of miR-146a in the neuropathogenesis of POCD and suggest that miR-146a may be a potential therapeutic target for POCD.

A comparison of effects of scalp nerve block and local anesthetic infiltration on inflammatory response, hemodynamic response, and postoperative pain in patients undergoing craniotomy for cerebral aneurysms: a randomized controlled trial.

BACKGROUND: The purpose of this study was to compare the effects of scalp nerve block (SNB) and local anesthetic infiltration (LA) with 0.75% ropivacaine on postoperative inflammatory response, intraoperative hemodynamic response, and postoperative pain control in patients undergoing craniotomy. METHODS: Fifty-seven patients were admitted for elective craniotomy for surgical clipping of a cerebral aneurysm. They were randomly divided into three groups: Group S (SNB with 15 mL of 0.75% ropivacaine), group I (LA with 15 mL of 0.75% ropivacaine) and group C (that only received routine intravenous analgesia). Pro-inflammatory cytokine levels in plasma for 72 h postoperatively, hemodynamic response to skin incision, and postoperative pain intensity were measured. RESULTS: The SNB with 0.75% ropivacaine not only decreased IL-6 levels in plasma 6 h after craniotomy but also decreased plasma CRP levels and increased plasma IL-10 levels 12 and 24 h after surgery compared to LA and routine analgesia. There were significant increases in mean arterial pressure 2 and 5 mins after the incision and during dura opening in Groups I and C compared with Group S. Group S had lower postoperative pain intensity, longer duration before the first dose of oxycodone, less consumption of oxycodone and lower incidence of PONV through 48 h postoperatively than Groups I and C. CONCLUSION: Preoperative SNB attenuated inflammatory response to craniotomy for cerebral aneurysms, blunted the hemodynamic response to scalp incision, and controlled postoperative pain better than LA or routine analgesia. TRIAL REGISTRATION: Clinicaltrials.gov NCT03073889 (PI:Xi Yang; date of registration:08/03/2017).

Comparative efficacy of individual renin-angiotensin system inhibitors on major renal outcomes in diabetic kidney disease: a network meta-analysis.

Background: Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) are two drug classes with well-documented renal protective effects. However, whether there is any difference among individual drugs remains unknown. In this study, we aimed to compare the efficacy of individual ACEIs/ARBs on major renal outcomes in adults with diabetic kidney disease (DKD). Methods: We conducted a Bayesian-framework network meta-analysis with a random effects model. We searched PubMed, Embase, Scopus, the Cochrane Central Register of Controlled Trials and ClinicalTrials.gov for clinical trials of ACEIs or ARBs as monotherapy compared with other conventional antihypertensive drugs or placebo. Primary outcomes were end-stage renal disease (ESRD) and albuminuria/proteinuria (including change in albuminuria/proteinuria, progression to macroalbuminuria and remission to normoalbuminuria). Secondary outcome was doubling of serum creatinine levels. We also assessed for hyperkalemia, cough and angioedema/edema. International prospective register of systematic reviews (PROSPERO) registration CRD42016036997. Results: A total of 100 studies with data for 22 365 DKD patients, the majority of whom had type 2 diabetes, were included. Individual ACEIs and ARBs at goal doses showed no significant differences in ESRD and doubling of serum creatinine levels. They also shared similar effects on albuminuria/proteinuria reduction and progression or remission of albuminuria. When combining three outcomes of albuminuria/proteinuria as a single endpoint, most ACEIs/ARBs consistently showed favorable antiproteinuric effect, with little difference in the possibility of being the superior treatment for improving albuminuria/proteinuria. Primary outcomes did not change substantially in meta-regressions and sensitivity analyses. Findings were limited by lack of dose equivalence and paucity of data for some outcomes. Conclusions: Based on the available evidence, individual ACEIs and ARBs at goal doses appeared to have no or little differences in their effect on major renal outcomes.

Identifying Phage Virion Proteins by Using Two-Step Feature Selection Methods.

Accurate identification of phage virion protein is not only a key step for understanding the function of the phage virion protein but also helpful for further understanding the lysis mechanism of the bacterial cell. Since traditional experimental methods are time-consuming and costly for identifying phage virion proteins, it is extremely urgent to apply machine learning methods to accurately and efficiently identify phage virion proteins. In this work, a support vector machine (SVM) based method was proposed by mixing multiple sets of optimal g-gap dipeptide compositions. The analysis of variance (ANOVA) and the minimal-redundancy-maximal-relevance (mRMR) with an increment feature selection (IFS) were applied to single out the optimal feature set. In the five-fold cross-validation test, the proposed method achieved an overall accuracy of 87.95%. We believe that the proposed method will become an efficient and powerful method for scientists concerning phage virion proteins.

Activation of cannabinoid receptor type 2 attenuates surgery-induced cognitive impairment in mice through anti-inflammatory activity.

BACKGROUND: Neuroinflammation plays a major role in postoperative cognitive dysfunction (POCD). Accumulated evidence indicates that cannabinoid receptor type 2 (CB2R) can mediate anti-inflammatory and immunomodulatory effects in part by controlling microglial activity. However, the impact of CB2R on postoperative cognition has not been investigated. We hypothesized that CB2R is involved in surgery-induced cognitive impairment in adult mice. METHODS: Adult C57BL/6 mice were subjected to intramedullary fixation surgery for tibial fracture under isoflurane anesthesia and CB2R agonist (JWH133) or CB2R antagonist (AM630) treatment. The mice were trained 24 h prior to surgery using a fear conditioning protocol and assessed in a novel context on postoperative days 1, 3, and 7 to evaluate cognitive function. Open-field testing was performed to evaluate the locomotor activity of the mice. The expression levels of IL-1ß, TNF-α, MCP-1, and CB2R in the hippocampus and prefrontal cortex were assessed by Western blotting; the expression of microglial marker CD11b in the CA1 area of the hippocampus and medial prefrontal cortex was assessed by immunostaining. RESULTS: The mice displayed no changes in locomotor activity after surgery and drug treatments. The mice exhibited impaired hippocampal-dependent memory accompanied by an increased expression of proinflammatory factors in the hippocampus and prefrontal cortex 1, 3, and 7 days after surgery, while hippocampal-independent memory remained unaffected at the same time points. JWH133 treatment attenuated surgery-induced memory loss, while AM630 treatment aggravated surgery-induced memory loss, paralleled by a decreased or increased expression of proinflammatory factors in the hippocampus and prefrontal cortex. The expression of CB2R in the hippocampus and prefrontal cortex was upregulated following surgery; however, it was downregulated by postoperative treatment with JWH133. Similarly, the expression of CD11b in the CA1 area of the hippocampus and medial prefrontal cortex was upregulated following surgery and downregulated by postoperative treatment with JWH133. CONCLUSIONS: These findings indicate that CB2R may modulate the neuroinflammatory and cognitive impairment in a mouse model of orthopedic surgery, and the activation of CB2R may effectively ameliorate the hippocampal-dependent memory loss of mice in the early postoperative stage.

Aerobic oxidative acylation of nitroarenes with arylacetic esters under mild conditions: facile access to diarylketones.

A facile and regioselective base-mediated aerobic oxidative acylation of nitroarenes to access diarylketones under mild conditions has been developed. It features the use of bench-stable and readily available arylacetates as acyl surrogates, and the absence of transition-metals and synthetic oxidants. This protocol involves a cascade CDC/oxidative decarboxylation process.

Short-Term Postoperative Cognitive Dysfunction and Inflammatory Response in Patients Undergoing Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study.

OBJECTIVES: To assess the association between short-term postoperative cognitive dysfuction (POCD) and inflammtory response in patients undergoing cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC). DESIGN: A prospective cohort study. SETTING: University medical centre. PARTICIPANTS: Fifty-one adult patients who had undergone CRS-HIPEC and twenty control participants. MEASUREMENTS: The inflammatory marker levels in plasma and cognitive function were measured. RESULTS: Twenty (39.2%, 20/51) patients developed POCD at 1 w after CRS-HIPEC. The patients with POCD had higher serum interleukin 1ß (IL-1ß), serum amyloid A (SAA), S100 calcium-binding protein ß (S-100ß), and high mobility group box-1 protein (HMGB-1) levels at 1 and 24 h postoperatively than patients without POCD. There was an association between POCD and the maximum IL-1ß and S-100ß concentrations in serum, which remained following adjustment for age and FBS. CONCLUSION: In this pilot study, perioperative inflammatory marker levels increase significantly after CRS-HIPEC in adult patients, and such elevations are associated with the development of short-term cognitive dysfunction after this complex surgery. These results suggested the need for a larger RCT to replicate and confirm these findings.

Toll-like receptor 4 knockout ameliorates neuroinflammation due to lung-brain interaction in mechanically ventilated mice.

Toll-like receptor 4 (TLR4) is a crucial receptor in the innate immune system, and increasing evidence supports its role in inflammation, stress, and tissue injury, including injury to the lung and brain. We aimed to investigate the effects of TLR4 on neuroinflammation due to the lung-brain interaction in mechanically ventilated mice. Male wild-type (WT) C57BL/6 and TLR4 knockout (TLR4 KO) mice were divided into three groups: (1) control group (C): spontaneous breathing; (2) anesthesia group (A): spontaneous breathing under anesthesia; and (3) mechanical ventilation group (MV): 6h of MV under anesthesia. The behavioral responses of mice were tested with fear conditioning tests. The histological changes in the lung and brain were assessed using hematoxylin-eosin (HE) staining. The level of TLR4 mRNA in tissue was measured using reverse transcription-polymerase chain reaction (RT-PCR). The levels of inflammatory cytokines were measured with an enzyme-linked immunosorbent assay (ELISA). Microgliosis, astrocytosis, and the TLR4 immunoreactivity in the hippocampus were measured by double immunofluorescence. MV mice exhibited impaired cognition, and this impairment was less severe in TLR4 KO mice than in WT mice. In WT mice, MV increased TLR4 mRNA expression in the lung and brain. MV induced mild lung injury, which was prevented in TLR4 KO mice. MV mice exhibited increased levels of inflammatory cytokines, increased microglia and astrocyte activation. Microgliosis was alleviated in TLR4 KO mice. MV mice exhibited increased TLR4 immunoreactivity, which was expressed in microglia and astrocytes. These results demonstrate that TLR4 is involved in neuroinflammation due to the lung-brain interaction and that TLR4 KO ameliorates neuroinflammation due to lung-brain interaction after prolonged MV. In addition, Administration of a TLR4 antagonist (100µg/mice) to WT mice also significantly attenuated neuroinflammation of lung-brain interaction due to prolonged MV. TLR4 antagonism may be a new and novel approach for the treatment and management of neuroinflammation in long-term mechanically ventilated patients.

Prolonged mechanical ventilation-induced neuroinflammation affects postoperative memory dysfunction in surgical mice.

INTRODUCTION: Patients undergoing surgery frequently develop neuropsychological disturbances, including cognitive decline or memory impairment, and routine clinical procedures such as mechanical ventilation (MV) may affect acute-phase brain outcome. We aimed to investigate the effect of the prolonged MV on postoperative memory dysfunction in surgical mice. METHODS: Male C57BL/6 mice were randomly divided into the following three groups: (1) The control group (group C) comprised anesthetized, unventilated animals; (2) the surgery group (subgroups S1h, S3h and S6h) was unventilated animals that underwent surgery under general anesthesia; and (3) the MV group (subgroups MV1h, MV3h and MV6h) was made up of animals under MV for 1 hour, 3 hours or 6 hours after surgery. Separate cohorts of animals were tested for memory function with fear conditioning tests or were killed at 6 hours, 1 day or 3 days postsurgery or post-MV to examine levels systemic and hippocampal interleukin (IL)-1ß, IL-6 and tumor necrosis factor α (TNFα), and assessed synaptic structure and microglial activation. Nuclear factor κB (NF-κB) p65, cytochrome c, cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP) activation were analyzed by Western blotting. RESULTS: The MV6h group showed increased CD11b-immunopositive cells, synapse degeneration, cytochrome c release, cleaved caspase-3 and cleaved PARP-1 activation after surgery, as well as a decrease in freezing time after surgery. At 6 hours and 1 day post-MV, MV6h increased NF-κB activation and levels of systemic and hippocampal IL-1ß, IL-6 and TNFα after surgery. CONCLUSIONS: Prolonged MV after surgery further aggravates cognitive decline that may stem from upregulation of hippocampal IL-1ß, IL-6 and TNFα, partially via activation of gliocytes in the surgical mouse hippocampus.

iRSpot-PseDNC: identify recombination spots with pseudo dinucleotide composition.

Meiotic recombination is an important biological process. As a main driving force of evolution, recombination provides natural new combinations of genetic variations. Rather than randomly occurring across a genome, meiotic recombination takes place in some genomic regions (the so-called 'hotspots') with higher frequencies, and in the other regions (the so-called 'coldspots') with lower frequencies. Therefore, the information of the hotspots and coldspots would provide useful insights for in-depth studying of the mechanism of recombination and the genome evolution process as well. So far, the recombination regions have been mainly determined by experiments, which are both expensive and time-consuming. With the avalanche of genome sequences generated in the postgenomic age, it is highly desired to develop automated methods for rapidly and effectively identifying the recombination regions. In this study, a predictor, called 'iRSpot-PseDNC', was developed for identifying the recombination hotspots and coldspots. In the new predictor, the samples of DNA sequences are formulated by a novel feature vector, the so-called 'pseudo dinucleotide composition' (PseDNC), into which six local DNA structural properties, i.e. three angular parameters (twist, tilt and roll) and three translational parameters (shift, slide and rise), are incorporated. It was observed by the rigorous jackknife test that the overall success rate achieved by iRSpot-PseDNC was >82% in identifying recombination spots in Saccharomyces cerevisiae, indicating the new predictor is promising or at least may become a complementary tool to the existing methods in this area. Although the benchmark data set used to train and test the current method was from S. cerevisiae, the basic approaches can also be extended to deal with all the other genomes. Particularly, it has not escaped our notice that the PseDNC approach can be also used to study many other DNA-related problems. As a user-friendly web-server, iRSpot-PseDNC is freely accessible at http://lin.uestc.edu.cn/server/iRSpot-PseDNC.

iTIS-PseTNC: a sequence-based predictor for identifying translation initiation site in human genes using pseudo trinucleotide composition.

Translation is a key process for gene expression. Timely identification of the translation initiation site (TIS) is very important for conducting in-depth genome analysis. With the avalanche of genome sequences generated in the postgenomic age, it is highly desirable to develop automated methods for rapidly and effectively identifying TIS. Although some computational methods were proposed in this regard, none of them considered the global or long-range sequence-order effects of DNA, and hence their prediction quality was limited. To count this kind of effects, a new predictor, called "iTIS-PseTNC," was developed by incorporating the physicochemical properties into the pseudo trinucleotide composition, quite similar to the PseAAC (pseudo amino acid composition) approach widely used in computational proteomics. It was observed by the rigorous cross-validation test on the benchmark dataset that the overall success rate achieved by the new predictor in identifying TIS locations was over 97%. As a web server, iTIS-PseTNC is freely accessible at http://lin.uestc.edu.cn/server/iTIS-PseTNC. To maximize the convenience of the vast majority of experimental scientists, a step-by-step guide is provided on how to use the web server to obtain the desired results without the need to go through detailed mathematical equations, which are presented in this paper just for the integrity of the new prection method.

Posttreatment with propofol attenuates lipopolysaccharide-induced up-regulation of inflammatory molecules in primary microglia.

BACKGROUND: Activation of microglia is involved in a broad range of neuroinflammatory diseases. Suppression of microglial activation may, therefore, contribute to alleviate the progression of neuroinflammatory diseases. It has been reported that propofol has a potent anti-inflammatory property. In the present study, we investigated the effects of posttreatment with propofol on the production of inflammatory molecules in lipopolysaccharide (LPS)-stimulated microglia. MATERIALS AND METHODS: Microglia were exposed to various concentrations (25, 50, 100, 250 µM) of propofol for 1 h after LPS stimulation for 24 h. The levels of proinflammatory mediators inducible nitric oxide synthase (iNOS)/nitric oxide (NO), cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) were measured. RESULTS: Propofol at a concentration of 25 µM did not affect the production of proinflammatory mediators, which was enhanced by LPS. At the concentrations of 50, 100, and 250 µM, propofol significantly inhibited LPS-mediated production of NO, PGE2, TNF-α, and IL-1ß and the expression of iNOSmRNA, COX-2mRNA, TNF-α mRNA, and IL-1ß mRNA. CONCLUSIONS: These results suggest that propofol, at clinically relevant concentrations, can reduce inflammatory responses in LPS-induced inflammation in activated microglia and might be an intravenous anesthetic of choice when patients with neuroinflammatory diseases require sedation and/or general anesthesia.