NETs Promote Inflammatory Injury by Activating cGAS-STING Pathway in Acute Lung Injury.

Acute respiratory distress syndrome (ARDS) threatens the survival of critically ill patients, the mechanisms of which are still unclear. Neutrophil extracellular traps (NETs) released by activated neutrophils play a critical role in inflammatory injury. We investigated the role of NETs and the underlying mechanism involved in acute lung injury (ALI). We found a higher expression of NETs and cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) in the airways, which was reduced by Deoxyribonuclease I (DNase I) in ALI. The administration of the STING inhibitor H-151 also significantly relieved inflammatory lung injury, but failed to affect the high expression of NETs in ALI. We isolated murine neutrophils from bone marrow and acquired human neutrophils by inducing HL-60 to differentiate. After the PMA interventions, exogenous NETs were obtained from such extracted neutrophils. Exogenous NETs intervention in vitro and in vivo resulted in airway injury, and such inflammatory lung injury was reversed upon degrading NETs with or inhibiting cGAS-STING with H-151 as well as siRNA STING. In conclusion, cGAS-STING participates in regulating NETs-mediated inflammatory pulmonary injury, which is expected to be a new therapeutic target for ARDS/ALI.

Adipose tissue alleviates the stress response by releasing adiponectin during laparoscopic surgery in patients with colorectal cancer.

BACKGROUND AND OBJECTIVES: Laparoscopic colorectal surgery causes a lower stress response than open surgery. Adiponectin is mainly derived from adipocytes and has antidiabetic, antioxidative, and anti-inflammatory capabilities. The objective of the present study was to investigate the protein expression of adiponectin in adipose tissue, and the serum levels of adiponectin, oxidative stress markers and proinflammatory factors during laparoscopic colorectal surgery and open surgery periods. METHODS: Forty patients aged 60 to 80, with American Society of Anesthesiologists (ASA) I ~ II who underwent radical resection of colorectal cancer were recruited to the study. Laparoscopic group and open group included 20 patients each. Mesenteric adipose tissue and venous blood before (T1) and at the end (T2) of surgery were collected to examine adiponectin levels, and venous blood was collected to examine serum levels of oxidative stress related markers (superoxide dismutase (SOD), glutathione (GSH), lipid peroxide (LPO), malondialdehyde (MDA)), and inflammation-related factors (interleukin (IL)-1ß, interleukin (IL)-6, tumor necrosis factor-α (TNF-α)). RESULTS: Protein and serum levels of adiponectin were analyzed, and adiponectin levels were significantly increased at T2 than T1 in the laparoscopic surgery, while adiponectin levels were significantly higher in the laparoscopic surgery than in the open surgery at T2. In addition, the serum levels of SOD and GSH were significantly higher in the laparoscopic surgery than in open surgery at T2. However, the serum levels of LPO, TNF-α, IL-1ß, and IL-6 were significantly lower in the laparoscopic group than in open group at T2. CONCLUSION: Laparoscopic surgery induced higher levels of adiponectin in both adipose tissue and the bloodstream. Oxidative stress and the inflammatory response were lower during laparoscopic colorectal surgery than during conventional open surgery. These data suggest that adipose tissue may alleviate the stress response during laparoscopic surgery by releasing adiponectin in patients with colorectal cancer.

Early enteral nutrition improves the outcome of critically ill patients with COVID-19: A retrospective study.

BACKGROUND AND OBJECTIVES: To evaluate the nutritional status of critically ill patients with COVID-19 and to determine which route of nutrition support is advantageous. METHODS AND STUDY DESIGN: This retrospective study was conducted in the ICU of a designated COVID-19 hospital. Patients were divided into an enteral nutrition (EN) group and parenteral nutrition (PN) group according to the initial route of nutrition support. NRS-2002 and NUTRIC were used to assess nutritional status. Blood nutritional markers such as albumin, total protein and hemoglobin were compared at baseline and seven days later. The primary endpoint was 28-day mortality. RESULTS: A total of 27 patients were enrolled in the study - 14 in the EN group and 13 in the PN group - and there were no significant demographic differences between groups. Most patients (96.3% NRS2002 score ≥5, 85.2% NUTRIC score ≥5) were at high nutritional risk. There was no significant difference in baseline albumin, total protein and hemoglobin levels between groups. After 7 days, albumin levels were significantly higher in the EN group than in the PN group (p=0.030). There was no significant difference in the other two indicators. The 28-day mortality was 50% in the EN group and 76.9% in the PN group. Kaplan-Meier survival analysis revealed significant differences between the groups (p=0.030). Cox proportional risk regression indicated that route of nutrition support was also an independent prognostic risk factor. CONCLUSIONS: The incidence of nutritional risk in critically ill patients with COVID-19 is very high. Early EN may be beneficial to patient outcomes.

Endothelial cell-specific anticoagulation reduces inflammation in a mouse model of acute lung injury.

Tissue factor (TF)-dependent coagulation contributes to lung inflammation and the pathogenesis of acute lung injury (ALI). In this study, we explored the roles of targeted endothelial anticoagulation in ALI using two strains of transgenic mice expressing either a membrane-tethered human tissue factor pathway inhibitor (hTFPI) or hirudin fusion protein on CD31+ cells, including vascular endothelial cells (ECs). ALI was induced by intratracheal injection of LPS, and after 24 h the expression of TF and protease-activated receptors (PARs) on EC in lungs were assessed, alongside the extent of inflammation and injury. The expression of TF and PARs on the EC in lungs was upregulated after ALI. In the two strains of transgenic mice, expression of either of hTFPI or hirudin by EC was associated with significant reduction of inflammation, as assessed by the extent of leukocyte infiltration or the levels of proinflammatory cytokines, and promoted survival after LPS-induced ALI. The beneficial outcomes were associated with inhibition of the expression of chemokine CCL2 in lung tissues. The protection observed in the CD31-TFPI-transgenic strain was abolished by injection of an anti-hTFPI antibody, but not by prior engraftment of the transgenic strains with WT bone marrow, confirming that the changes observed were a specific transgenic expression of anticoagulants by EC. These results demonstrate that the inflammation in ALI is TF and thrombin dependent, and that expression of anticoagulants by EC significantly inhibits the development of ALI via repression of leukocyte infiltration, most likely via inhibition of chemokine gradients. These data enhance our understanding of the pathology of ALI and suggest a novel therapeutic strategy for treatment.

Napabucasin Attenuates Resistance of Breast Cancer Cells to Tamoxifen by Reducing Stem Cell-Like Properties.

BACKGROUND Tamoxifen (TAM) is the first-line drug for estrogen receptor-positive (ER+) breast cancer (BC) treatment. However, its resistance is a main obstacle in clinical practice. Thus, new therapeutic agents are urgently needed to fight TAM resistance. MATERIAL AND METHODS Here, we constructed TAM-resistant ER+BC cells with TAM resistance, named MCF-7-R. Western blot, quantitative real-time PCR (qRT-PCR), ALDH1 activity analysis, and spheroid-forming detection were used to detect the stemness of cells and the effects of napabucasin (NP) on BC cell stemness. Cell counting kit-8 (CCK8) assay was used to evaluate the effects of NP on cell viability. RESULTS MCF-7-R cells exhibited higher stemness compared with the parental MCF-7 cells, which was evident by the increased spheroid formation ability at diluted concentration, aldehyde dehydrogenase (ALDH) activity, and expression of stemness critical biomarkers (Oct4, Nanog, and Sox2). Additionally, it was found that napabucasin (NP) specifically killed MCF-7-T cells, characterized by remarkably decreased IC50 value. Notably, NP reduced MCF-7-R cell stemness, which was evident as the decreased stemness marker expression, spheroid-forming capacity, and ALDH1 activity. Importantly, NP attenuated TAM resistance of MCF-7-R cells and enhanced sensitivity of MCF-7 cells to TAM. Mechanistic study showed that NP inhibited STAT3 activation, and overexpression of STAT3 rescued NP-mediated inhibition of the stemness-like characteristics of MCF-7-R cells. CONCLUSIONS NP might be used as an adjuvant therapy for ER+ BC patients with TAM resistance.

The Cardioprotective Effects of Remote Ischemic Conditioning in a Rat Model of Acute Myocardial Infarction.

BACKGROUND Cardiac remote ischemic conditioning (RIC) is a noninvasive cardioprotective method in ischemia-reperfusion injury and acute myocardial infarction (AMI). The aims of this study were to investigate the effects of RIC in a rat model of AMI. MATERIAL AND METHODS Adult male Sprague-Dawley rats included the AMI group that underwent ligation of the left anterior descending (LAD) coronary artery (n=24), the RIC group that consisted the AMI rat model treated with RIC once daily in the left hind limb until days 1, 7 and 14 (n=24), and the sham group (n=24). Myocardial infarct size was measured by routine histology with triphenyltetrazolium chloride (TTC) and Masson's trichrome histochemical staining for myocardial necrosis and fibrosis, respectively. Serum levels of Bcl-2, Bax, caspase-3, and inducible nitric oxide synthase (iNOS) were measured by enzyme-linked immunosorbent assay (ELISA). The apoptosis index was detected using the TUNEL assay. Spectrophotometry of the myocardium was used to identify mitochondrial complexes and myocardial ATP. RESULTS The RIC group showed improved cardiac hemodynamics, reduced the size of the myocardial infarction, upregulated expression of Bcl-2, and down-regulation of the levels of Bax, caspase-3, and iNOS, and reduced cardiac myocyte apoptosis and inhibited the opening of the mitochondrial permeability transition pore (MPTP). CONCLUSIONS In a rat model of AMI, RIC improved the hemodynamic index, reduce the levels of apoptosis and myocardial injury, and improved mitochondrial function.

Efficacy and safety of benralizumab for eosinophilic asthma: A systematic review and meta-analysis of randomized controlled trials.

CONTEXT: Benralizumab is a humanized monoclonal antibody that targets the α chain of the IL-5 receptor (IL-5Rα) and is currently being assessed in clinical trials for asthma control. OBJECTIVE: Our systematic review and meta-analysis intends to evaluate the therapeutic efficacy and safety of benralizumab in patients with eosinophilic asthma. DATA SOURCES AND EXTRACTION: Literature searches of PubMed, Embase, and the Cochrane Library were performed to identify randomized controlled trials of benralizumab and clinic outcomes in asthmatics. RESULTS: In total, 7 articles with 2,321 subjects met our inclusion criteria. From this pooled analysis, we found that benralizumab significantly reduces exacerbations (RR: 0.63, 95% CI: 0.52-0.76, p < 0.00001; I2 = 52%, p = 0.06) compared to placebo in eosinophilic asthma. There was no statistical trend for improvement in forced expiratory volume in 1 second or asthma control indices such as Quality of Life Assessment (AQLQ) and Asthma Control Questionnaire score in benralizumab-treated patients. In addition, safety data indicated that benralizumab administration resulted no increasing incidence of adverse events and was well tolerated (RR: 1.00, 95% CI: 0.95-1.05, p = 0.96; I2 = 40%, p = 0.13). CONCLUSION: These results demonstrate the efficacy and safety of benralizumab for asthma patients with severe or uncontrolled symptoms and elevated eosinophils and provide support for benralizumab as an ideal option to treat asthma in this patient population.

Therapeutic effects of a taurine-magnesium coordination compound on experimental models of type 2 short QT syndrome.

Short QT syndrome (SQTS) is a genetic arrhythmogenic disease that can cause malignant arrhythmia and sudden cardiac death. The current therapies for SQTS have application restrictions. We previously found that Mg· (NH2CH2CH2SO3)2· H2O, a taurine-magnesium coordination compound (TMCC) exerted anti-arrhythmic effects with low toxicity. In this study we established 3 different models to assess the potential anti-arrhythmic effects of TMCC on type 2 short QT syndrome (SQT2). In Langendorff guinea pig-perfused hearts, perfusion of pinacidil (20 µmol/L) significantly shortened the QT interval and QTpeak and increased rTp-Te (P<0.05 vs control). Subsequently, perfusion of TMCC (1-4 mmol/L) dose-dependently increased the QT interval and QTpeak (P<0.01 vs pinacidil). TMCC perfusion also reversed the rTp-Te value to the normal range. In guinea pig ventricular myocytes, perfusion of trapidil (1 mmol/L) significantly shortened the action potential duration at 50% (APD50) and 90% repolarization (APD90), which was significantly reversed by TMCC (0.01-1 mmol/L, P<0.05 vs trapidil). In HEK293 cells that stably expressed the outward delayed rectifier potassium channels (IKs), perfusion of TMCC (0.01-1 mmol/L) dose-dependently inhibited the IKs current with an IC50 value of 201.1 µmol/L. The present study provides evidence that TMCC can extend the repolarization period and inhibit the repolarizing current, IKs, thereby representing a therapeutic candidate for ventricular arrhythmia in SQT2.

Ultrasensitive and Multifunction Plasmonic Temperature Sensor with Ethanol-Sealed Asymmetric Ellipse Resonators.

In order to improve the low temperature sensitivity of conventional sensors, a plasmonic multifunction temperature sensor with high sensitivity is proposed and investigated systematically in this paper. The sensor consists of two metal layers and two ethanol-sealed elliptical resonators connected to a straight waveguide by two rectangular tubes. We numerically analyzed the transmission characteristics of the Nano-device to assess its performance with the finite element method and achieved great optical properties. The results show that an obvious blue shift of the transmission spectrum appears by varying temperatures, exhibiting a great sensing effect. Sensitivity of the sensor reaches -3.64 nm/°C, far greater than conventional temperature sensors. Our research also demonstrates that the transmission spectrum could be modulated efficiently by the ratio of semi-short axis to semi-major axis of the ellipse resonators and the width of two same rectangular tubes. Furthermore, the Nano-device has a filtering characteristic. The transmittances of pass-band and stop-band are 96.1% and 0.1%, respectively. The results of this study can pave the way for low-cost sensing application in high-density photonic circuits and biosensors.

A preoperative nomogram predicts prognosis of up front resectable patients with pancreatic head cancer and suspected venous invasion.

BACKGROUND: Pancreatic head adenocarcinoma is commonly diagnosed at an advanced stage when adjacent vascular invasion is present. This study aimed to establish a preoperative prognostic nomogram for patients who underwent attempted curative resectional surgery for pancreatic head cancer with suspected peripancreatic venous invasion. METHODS: Data on all consecutive patients were retrospectively collected from 2012 to 2016 at four academic institutions. The demographic and radiological parameters were analyzed using univariate and multivariate Cox regression analyses. The final nomogram was established using the concordance Harrell's C-indices and calibration curves from data obtained in three institutions and validated in the cohort of patients coming from the fourth institution. RESULTS: The nomogram was constructed using data from 178 patients while the validation cohort consisted of 61 patients. Age, length of tumor contact, peripancreatic venous abnormalities and lymph node staging were independent factors of overall survival. The nomogram showed good probabilities of survival on calibration curves. The C-index of the model in predicting overall survival (OS) was 0.824 for the validation cohort. CONCLUSIONS: The nomogram accurately predicted OS in patients with pancreatic head cancer with suspected peripancreatic venous invasion after attempted curative pancreatic resectional surgery.

Effects of non-invasive remote ischemic conditioning on rehabilitation after myocardial infarction.

Recent studies have demonstrated that remote ischemic conditioning (RIC) creates cardioprotection against ischemia/reperfusion injury and myocardial infarction (MI); however, the effects of non-invasive remote ischemic conditioning (nRIC) on prognosis and rehabilitation after MI (post-MI) remain unknown. We successfully established MI models involving healthy adult male Sprague-Dawley rats. The nRIC group repeatedly underwent 5 min of ischemia and 5 min of reperfusion in the left hind limb for three cycles every other day until weeks 4, 6, and 8 after MI. nRIC improved cardiac hemodynamic function and mitochondrial respiratory function through increasing myocardial levels of mitochondrial respiratory chain complexes I, II, III, IV, and adenosine triphosphate (ATP) and decreasing the activity of nitric oxide synthase (NOS). nRIC could inhibit cardiomyocytes apoptosis and reduce myocardium injury through raising the expression of Bcl-2 and reduced the content of creatine kinase-MB, cardiac troponin I and Bax. The results indicated that long-term nRIC could accelerate recovery and improve prognosis and rehabilitation in post-MI rats.

Chemical Constituents from Euphorbia kansui.

In this research, a new triterpenoid, tirucalla-8,24-diene-3ß,11ß-diol-7-one (1), and eupha-8,24-diene-3ß,11ß-diol-7-one (2), which was isolated from Euphorbia kansui for the first time, together with twelve other known compounds (3-14), were isolated from the ethyl acetate extract of Euphorbia kansui. Their structures were elucidated based on High resolution electrospray ionization mass spectrometry (HR-ESI-MS), Infrared Spectroscopy (IR), 1D and 2D Nuclear Magnetic Resonance (NMR) data. Both constituents 1 and 2 exhibited moderate cytotoxicity against colon cancer HCT-116, gastric cancer MKN-45 and breast cancer MCF-7.

[Comparative study on acute toxicity of different active ingredient fractions from Kansui stir-baked with vinegar on zebrafish embryos].

The 24 h normal developing zebrafish embryos were used to evaluate the acute toxicity and the compounds of respective fractions were analyzed by UFLC-Q-TOF-MS simultaneously. Nine concentration groups with respective concentration and a blank control group were designed for each fraction to investigate their effect on survival rates of zebrafish embryos 96 h after drug administration, and calculate the median lethal concentration (LC50) of different fractions to zebrafish embryos. The results showed that all of the fractions had acute toxicity to zebrafish embryos except VEKD, and the order was as follows: VEKB, VEKC, VEKA and VEKD. According to the results of UFLC-Q-TOF-MS, the chemical ingredients contained in VEKB and VEKC were mainly composed of ingenane-type and japhane-type diterpenoids, respectively. It could be speculated that japhane-type diterpenoids might be the active compounds with lower toxicity associated with the results of toxicity study, providing some references for the further research on effective material basis of Kansui stir-baked with vinegar according to the principle of "drastic medicine, no death risks".

Salvage surgery for neck residue or recurrence of nasopharyngeal carcinoma after primary radiotherapy: options of surgical methods and regions.

BACKGROUND: Salvage surgery has been recommended as the approach of choice for neck residue or recurrence of nasopharyngeal carcinoma (NPC) after primary radiotherapy (RT). This study aimed to assess the outcome and prognostic factors, options for different surgical methods, and the extent of neck dissection (ND) for patients. METHODS: NPC patients who had undergone RT and received salvage surgery for neck residue or recurrence from January 2001 to December 2011 were retrospectively analyzed. The overall survival (OS) rate was calculated by Kaplan-Meier method, and prognostic factors were determined by log-rank test and Cox regression analysis. RESULTS: In 153 cases, 96 cases have level I dissections. The metastasis rate was 20/153 (13.07%) for level I metastasis and 7/153 (4.58%) for parotid gland cases. The 3- and 5-year OS rate was 57.2 and 40.6%, respectively, and median survival time was 49 months. By univariate analysis, the age, rN staging, size of lymph nodes (LN), extra-capsular spread (ECS), and surgical procedure were significant prognostic factors. By multivariable analysis, the age, rN staging, and size of LN were significant prognostic factors. CONCLUSIONS: Salvage surgery is effective for neck failure of NPC after primary treatment, but patients with age >50 years, stage rN3, or LN >6 cm have poor prognosis.

Exploring competencies: a qualitative study of Chinese nurse managers.

AIM: To identify core competencies needed in the transition of nurse managers on the way to excellence. BACKGROUND: There is growing recognition of the importance of nurse managers in hospitals. Most managers still learn through their failures and few studies have described the perceptions of nurse managers in China. It is vital to understand what competencies Chinese nurse managers should have in order to establish suitable training programmes and improve their management skills. METHOD: A phenomenological approach that included in-depth interviews with 12 nurse managers in six Chinese hospitals was conducted. RESULTS: The transition to management included four phases: the adaptive phase, the running-in and stable phase, the stagnation phase and the maturation phase. CONCLUSION: In order to fulfil their clinical responsibilities, nurse managers need to develop multifaceted competencies, specifically in communication and stress management. Ideally, nurse managers should progress through the four phases mentioned above to achieve excellence. IMPLICATIONS: There is a requirement for utilising various methods for nurse managers in adapting new roles, improving communication and relieving stress.

Anisotropic shock sensitivity in a single crystal δ-cyclotetramethylene tetranitramine: a reactive molecular dynamics study.

The anisotropic shock sensitivity in a single crystal δ-cyclotetramethylene tetranitramine (δ-HMX) was investigated using the compress-shear reactive dynamics (CS-RD) computational protocol. Significant anisotropies in the thermo-mechanical and chemical responses were found by measuring the shear stress, energy, temperature, and chemical reactions during the dynamical process for the shock directions perpendicular to the (100), (010), (001), (110), (101), (011), and (111) planes. We predict that δ-HMX is sensitive for the shocks perpendicular to the (111), (011), (110), and (101) planes, which is intermediate to the (100) and (010) plane and is insensitive to the (001) plane. The internal energy accumulated within the duration of the surmounting shear stress barrier is a useful criterion to distinguish the sensitive directions from the less sensitive ones. The molecular origin of the anisotropic sensitivity is suggested to be the intermolecular steric arrangements across a slip plane induced by shock compression. The shear deformation induced by the shock along the sensitive direction encounters strong intermolecular contacts and has small intermolecular free space for geometry relaxation when the molecules collide, leading to high shear stress barriers and energy accumulation, which benefits the temperature increase and initial chemical bond breaking that trigger further reactions.

Biological evaluation of a cytotoxic 2-substituted benzimidazole copper(II) complex: DNA damage, antiproliferation and apoptotic induction activity in human cervical cancer cells.

Exploring novel chemotherapeutic agents is a great challenge in cancer medicine. To that end, 2-substituted benzimidazole copper(II) complex, [Cu(BMA)Cl2]·(CH3OH) (1) [BMA = N,N'-bis(benzimidazol-2-yl-methyl)amine], was synthesized and its cytotoxicity was characterized. The interaction between complex 1 and calf thymus DNA was detected by spectroscopy methods. The binding constant (K b = 1.24 × 10(4 )M(-1)) and the apparent binding constant (K app = 6.67 × 10(6 )M(-1)) of 1 indicated its moderate DNA affinity. Complex 1 induced single strand breaks of pUC19 plasmid DNA in the presence of H2O2 through an oxidative pathway. Cytotoxicity studies proved that complex 1 could inhibit the proliferation of human cervical carcinoma cell line HeLa in both time- and dose-dependent manners. The results of nuclei staining by Hoechst 33342 and alkaline single-cell gel electrophoresis proved that complex 1 caused cellular DNA damage in HeLa cells. Furthermore, treatment of HeLa cells with 1 resulted in S-phase arrest, loss of mitochondrial potential, and up-regulation of caspase-3 and -9 in HeLa cells, suggesting that complex 1 was capable of inducing apoptosis in cancer cells through the intrinsic mitochondrial pathway.

Hemorrhagic cystitis in gastric cancer after nanoparticle albumin-bound paclitaxel: A case report.

BACKGROUND: The advanced first-line regimen for advanced gastric cancer is based on a combination of fluoropyrimidine and platinum and/or paclitaxel (PTX), forming a two- or three-drug regimen. Compared to conventional PTX, nanoparticle albumin-bound PTX (Nab-PTX) has better therapeutic effects and fewer adverse effects reported in studies. Nab-PTX is a great option for patients presenting with advanced gastric cancer. Herein, we highlight an adverse event (hemorrhagic cystitis) of Nab-PTX in advanced gastric cancer. CASE SUMMARY: A 55-year-old male was diagnosed with lymph node metastasis after a laparoscopic-assisted radical gastrectomy for gastric cancer that was treated by Nab-PTX and S-1 (AS). On the 15th day after treatment with AS, he was diagnosed with hemorrhagic cystitis. CONCLUSION: Physicians should be aware that hemorrhagic cystitis is a potential adverse event associated with Nab-PTX treatment.

Microbial community of municipal drinking water in Hangzhou using metagenomic sequencing.

While traditional culture-dependent methods can effectively detect certain microorganisms, the comprehensive composition of the municipal drinking water (DW) microbiome, including bacteria, archaea, and viruses, remains unknown. Metagenomic sequencing has opened the door to accurately determine and analyze the entire microbial community of DW, providing a comprehensive understanding of DW species diversity, especially in the context of public health concerns during the COVID-19 era. In this study, we found that most of the culturable bacteria and some fecal indicator bacteria, such as Escherichia coli and Pseudomonas aeruginosa, were non-culturable using culture-dependent methods in all samples. However, metagenomic analysis showed that the predominant bacterial species in the DW samples belonged to the phyla Proteobacteria and Planctomycetes. Notably, the genus Methylobacterium was the most abundant in all water samples, followed by Sphingomonas, Gemmata, and Azospirilum. While low levels of virulence-associated factors, such as the Esx-5 type VII secretion system (T7SS) and DevR/S, were detected, only the erythromycin resistance gene erm(X), an rRNA methyltransferase, was identified at low abundance in one sample. Hosts corresponding to virulence and resistance genes were identified in some samples, including Mycobacterium spp. Archaeal DNA (Euryarchaeota, Crenarchaeota) was found in trace amounts in some DW samples. Viruses such as rotavirus, coxsackievirus, human enterovirus, and SARS-CoV-2 were negative in all DW samples using colloidal gold and real-time reverse transcription polymerase chain reaction (RT‒PCR) methods. However, DNA encoding a new order of reverse-transcribing viruses (Ortervirales) and Herpesvirales was found in some DW samples. The metabolic pathways of the entire microbial community involve cell‒cell communication and signal secretion, contributing to cooperation between different microbial populations in the water. This study provides insight into the microbial community and metabolic process of DW in Hangzhou, China, utilizing both culture-dependent methods and metagenomic sequencing combined with bioinformatics tools during the COVID-19 pandemic era.

Distinct binding conformations of epinephrine with α- and ß-adrenergic receptors.

Agonists targeting α2-adrenergic receptors (ARs) are used to treat diverse conditions, including hypertension, attention-deficit/hyperactivity disorder, pain, panic disorders, opioid and alcohol withdrawal symptoms, and cigarette cravings. These receptors transduce signals through heterotrimeric Gi proteins. Here, we elucidated cryo-EM structures that depict α2A-AR in complex with Gi proteins, along with the endogenous agonist epinephrine or the synthetic agonist dexmedetomidine. Molecular dynamics simulations and functional studies reinforce the results of the structural revelations. Our investigation revealed that epinephrine exhibits different conformations when engaging with α-ARs and ß-ARs. Furthermore, α2A-AR and ß1-AR (primarily coupled to Gs, with secondary associations to Gi) were compared and found to exhibit different interactions with Gi proteins. Notably, the stability of the epinephrine-α2A-AR-Gi complex is greater than that of the dexmedetomidine-α2A-AR-Gi complex. These findings substantiate and improve our knowledge on the intricate signaling mechanisms orchestrated by ARs and concurrently shed light on the regulation of α-ARs and ß-ARs by epinephrine.