Upstream stimulating factor 2 aggravates neuropathic pain induced in spinal nerve ligation-induced mice via regulating SNHG5/miR-181b-5p.

BACKGROUND: Upstream stimulating factor 2 (USF2) belongs to basic-Helix-Loop-Helix-Leucine Zipper transcription factor family, regulating expression of genes involved in immune response or energy metabolism network. Role of USF2 in neuropathic pain was evaluated. METHODS: Mice were intraspinally injected with adenovirus for knockdown of USF2 (Ad-shUSF2), and then subjected to spinal nerve ligation (SNL) to induce neuropathic pain. Distribution and expression of USF2 was detected by western blot and immunofluorescence. Mechanical and thermal pain sensitivity were examined by paw withdrawal thresholds (PWT) and paw withdrawal latency (PWL). Chromatin immunoprecipitation (ChIP) and luciferase activity assays were performed to detect binding ability between USF2 and SNHG5. RESULTS: The expression of USF2 was elevated and colocalized with astrocytes and microglia in L5 dorsal root ganglion (DRG) of SNL-induced mice. Injection of Ad-shUSF2 attenuated SNL-induced decrease of PWT and PWL in mice. Knockdown of USF2 increased level of IL-10, but decreased TNF-α, IL-1ß, and IL-6 in SNL-induced mice. Silence of USF2 enhanced protein expression of CD206, while reduced expression of CD16 and CD32 in SNL-induced mice. USF2 bind to promoter of SNHG5, and weakened SNL-induced up-regulation of SNHG5. SNHG5 bind to miR-181b-5p, and miR-181b-5p to interact with CXCL5. CONCLUSION: Silence of USF2 ameliorated neuropathic pain, suppressed activation of M1 microglia and inhibited inflammation in SNL-induced mice through regulation of SNHG5/miR-181b-5p/CXCL5 axis. Therefore, USF2/SNHG5/miR-181b-5p/CXCL5 might be a promising target for neuropathic pain. However, the effect of USF2/SNHG5/miR-181b-5p/CXCL5 on neuropathic pain should also be investigated in further research.

Phase III clinical trial comparing the efficacy and safety of adamgammadex with sugammadex for reversal of rocuronium-induced neuromuscular block.

BACKGROUND: Preliminary clinical trials of adamgammadex, a new cyclodextrin-based selective reversal agent, have demonstrated its efficacy in reversing neuromuscular block by rocuronium. METHODS: This multicentre, randomised, double-blind, positive-controlled, non-inferiority phase III clinical trial compared the efficacy and safety of adamgammadex and sugammadex. We randomised 310 subjects to receive adamgammadex (4 mg kg-1) or sugammadex (2 mg kg-1) at reappearance of the second twitch of the train-of-four (TOF), and standard safety data were collected. RESULTS: For the primary outcome, the proportion of patients with TOF ratio ≥0.9 within 5 min was 98.7% in the adamgammadex group vs 100% in the sugammadex group, with a point estimate and 95% confidence interval (CI) of 1.3% (-4.6%, +1.3%); the lower limit was greater than the non-inferiority margin of -10%. For the key secondary outcome, the median (inter quartile range) time from the start of administration of adamgammadex or sugammadex to recovery of TOF ratio to 0.9 was 2.25 (1.75, 2.75) min and 1.75 (1.50, 2.00) min, respectively. The difference was 0.50 (95% CI: 0.25, 0.50); the upper limit was lower than the non-inferiority margin of 5 min. In addition, there were no inferior results observed in secondary outcomes. Adamgammadex had a lower incidence of adverse drug reactions compared with sugammadex (anaphylactic reaction, recurarisation, decreased heart rate, and laryngospasm; P=0.047). CONCLUSIONS: Adamgammadex was non-inferior to sugammadex with a possible lower incidence of adverse drug reactions compared with sugammadex. Adamgammadex may have a potential advantage in terms of its overall risk-benefit profile. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR2000039525. Registered October 30, 2020. https://www.chictr.org.cn/showproj.html?proj=56825.

Effect of volatile versus propofol anaesthesia on major complications and mortality after cardiac surgery: a multicentre randomised trial.

BACKGROUND: The comparative effectiveness of volatile anaesthesia and total intravenous anaesthesia (TIVA) in terms of patient outcomes after cardiac surgery remains a topic of debate. METHODS: Multicentre randomised trial in 16 tertiary hospitals in China. Adult patients undergoing elective cardiac surgery were randomised in a 1:1 ratio to receive volatile anaesthesia (sevoflurane or desflurane) or propofol-based TIVA. The primary outcome was a composite of predefined major complications during hospitalisation and mortality 30 days after surgery. RESULTS: Of the 3123 randomised patients, 3083 (98.7%; mean age 55 yr; 1419 [46.0%] women) were included in the modified intention-to-treat analysis. The composite primary outcome was met by a similar number of patients in both groups (volatile group: 517 of 1531 (33.8%) patients vs TIVA group: 515 of 1552 (33.2%) patients; relative risk 1.02 [0.92-1.12]; P=0.76; adjusted odds ratio 1.05 [0.90-1.22]; P=0.57). Secondary outcomes including 6-month and 1-yr mortality, duration of mechanical ventilation, length of ICU and hospital stay, and healthcare costs, were also similar for the two groups. CONCLUSIONS: Among adults undergoing cardiac surgery, we found no difference in the clinical effectiveness of volatile anaesthesia and propofol-based TIVA. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR-IOR-17013578).

Identification of ferroptosis-related genes in male mice with sepsis-induced acute lung injury based on transcriptome sequencing.

BACKGROUND: Sepsis can result in acute lung injury (ALI). Studies have shown that pharmacological inhibition of ferroptosis can treat ALI. However, the regulatory mechanisms of ferroptosis in sepsis-induced ALI remain unclear. METHODS: Transcriptome sequencing was performed on lung tissue samples from 10 sepsis-induced mouse models of ALI and 10 control mice. After quality control measures, clean data were used to screen for differentially expressed genes (DEGs) between the groups. The DEGs were then overlapped with ferroptosis-related genes (FRGs) to obtain ferroptosis-related DEGs (FR-DEGs). Subsequently, least absolute shrinkage and selection operator (Lasso) and Support Vector Machine-Recursive Feature Elimination (SVM-RFE) were used to obtain key genes. In addition, Ingenuity Pathway Analysis (IPA) was employed to explore the disease, function, and canonical pathways related to the key genes. Gene set enrichment analysis (GSEA) was used to investigate the functions of the key genes, and regulatory miRNAs of key genes were predicted using the NetworkAnalyst and StarBase databases. Finally, the expression of key genes was validated with the GSE165226 and GSE168796 datasets sourced from the Gene Expression Omnibus (GEO) database and using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Thirty-three FR-DEGs were identified between 1843 DEGs and 259 FRGs. Three key genes, Ncf2, Steap3, and Gclc, were identified based on diagnostic models established by the two machine learning methods. They are mainly involved in infection, immunity, and apoptosis, including lymphatic system cell migration and lymphocyte and T cell responses. Additionally, the GSEA suggested that Ncf2 and Steap3 were similarly enriched in mRNA processing, response to peptides, and leukocyte differentiation. Furthermore, a key gene-miRNA network including 2 key genes (Steap3 and Gclc) and 122 miRNAs, and a gene-miRNA network with 1 key gene (Steap3) and 3 miRNAs were constructed using NetworkAnalyst and StarBase, respectively. Both databases predicted that mmu-miR-15a-5p was the target miRNA of Steap3. Finally, Ncf2 expression was validated using both datasets and qRT-PCR, and Steap3 was validated using GSE165226 and qRT-PCR. CONCLUSIONS: This study identified two FR-DEGs (Ncf2 and Steap3) associated with sepsis-induced ALI via transcriptome analyses, as well as their functional and metabolic pathways.

Efficacy and safety of ciprofol vs. propofol for the induction and maintenance of general anaesthesia: A multicentre, single-blind, randomised, parallel-group, phase 3 clinical trial.

BACKGROUND: HSK3486 (ciprofol) is a 2,6-disubstituted phenol derivative that acts like propofol as an agonist at the gamma-aminobutyric acid-A (GABA A ) receptor. OBJECTIVE: To investigate the efficacy and safety of HSK3486 for general anaesthesia induction and maintenance. DESIGN: A single-blinded, randomised, parallel-group, phase 3 trial. SETTING: Involving 10 study centres, from November 24, 2020 to January 25, 2021. PATIENTS: A total of 129 patients undergoing nonemergency, noncardiothoracic, and nonneurosurgical elective surgery. INTERVENTION: Patients were randomly assigned at a 2:1 ratio into HSK3486 or propofol groups, to receive HSK3486 (0.4 mg kg -1 ) or propofol (2.0 mg kg -1 ) for induction before a maintenance infusion at initial rates of 0.8 and 5.0 mg kg -1 h -1 , and were adjusted to maintain a bispectral index (BIS) of 40-60 until the end of surgery. MAIN OUTCOME MEASURES: Noninferiority between the drugs was evaluated as the lower limit of the 95% confidence interval (CI) for the between-group difference in the success rate of anesthetic maintenance (primary outcome) >-8%. Secondary outcomes included successful anaesthetic induction, full alertness and spontaneous breathing recovery, time until leaving the postanaesthesia care unit and changes in BIS. Safety profiles were also measured. RESULTS: Of 129 enrolled patients, 128 completed the trial, with 86 in the HSK3486 group and 42 in the propofol group. The success rate for the maintenance of general anaesthesia was 100% for both groups, and noninferiority of HSK3486 was confirmed (95% CI -4.28% to 8.38%). No significant differences were found between the two groups of patients with regard to secondary outcomes (all P  > 0.05). There appeared to be a comparable incidence of treatment for emergency adverse events (TEAEs) (80.2% vs. 81.0%, P  = 1.000) and drug-related TEAEs (57.0% vs. 64.3%, P  = 0.451) in the HSK3486 and propofol groups. CONCLUSION: HSK3486 had a noninferior efficacy profile compared to propofol, exhibiting excellent tolerance. TRIAL REGISTRATION: Clinicaltrials.gov, identifier: NCT04511728.

Rapid Detection of Strawberry Mild Yellow Edge Virus with a Lateral Flow Strip Reverse Transcription Recombinase Polymerase Amplification Assay.

Strawberry mild yellow edge virus (SMYEV) is a latent virus that severely affects the yield and quality of strawberry fruit. The technology suitable for rapid and accurate detection of SMYEV on site is important to effectively control its spread. In this study, a reverse transcription recombinase polymerase amplification combined with lateral flow strip (SMYEV-RT-RPA-LF), targeting the conserved genome of Beijing SMYEV isolate, was established to diagnose SMYEV in strawberries. The SMYEV-RT-RPA-LF assay showed no cross-reaction with other strawberry viruses. The sensitivity of SMYEV-RT-RPA-L assay was 100 times higher than that of RT-PCR (10 pg/µL). In addition, through the detection of suspected samples in the field, it was found that the accuracy of SMYEV-RT-RPA-L assay was consistent with the RT-PCR results. However, compared with RT-PCR, SMYEV-RT-RPA-LF assay has the advantages of simple operation, time savings, and high specificity and sensitivity, indicating the potential application of SMYEV-RT-RPA-LF in the rapid field diagnosis of SMYEV.

Effect of different doses of esketamine compared with fentanyl combined with propofol on hypotension in patients undergoing painless abortion surgery: a prospective, randomized, double-blind controlled clinical trial.

BACKGROUND: Opioids analgesics commonly used in abortion procedures are associated with respiratory and circulatory depression. Esketamine is a N-methyl-D-aspartate receptor (NMDA) antagonist and a common analgesic. The drug has several advantages including rapid onset and offset and it causes minimal cardiorespiratory depression. However, studies have not explored the effects of esketamine in patients undergoing painless abortion surgery. Therefore, the present study sought to evaluate the effect of different doses of esketamine compared with the effect of fentanyl on incidence of perioperative hypotension in patients undergoing painless abortion surgery and to explore the optimal esketamine dose for this population. METHODS: A total of 178 female patients undergoing painless abortion surgery were enrolled to the current study. The patients were aged 18-45 years, had a body mass index (BMI) of 18-28 kg m- 2 and a class I or II physical status as determined using the American Society of Anesthesiologists (ASA) system. Patients were randomly assigned to four groups as follows: group F (n = 45) in which patients underwent intravenous (IV) administration of 1 µg kg- 1 fentanyl followed by IV administration of 2 mg kg- 1 propofol, and group EL, group EM and group EH (n = 45, 44, 44) with patients receiving IV administration of 0.2 mg kg- 1, 0.25 mg kg- 1, 0.3 mg kg- 1 esketamine, respectively, followed by IV administration of 2 mg kg- 1 propofol. The primary outcome of the study was the incidence of hypotension whereas secondary outcomes included incidence of adverse events, perioperative changes of vital signs, anesthesia induction time, recovery time and dischargeable time, propofol addition, as well as patient, surgeon and anesthesiologist satisfaction levels. RESULTS: The findings showed that the incidence of hypotension was significantly lower in subjects in group EL, group EM and group EH (0, 0, 0%) relative to the incidence in patients in group F (20%) (χ2 = 19.648; P = 0.000). In this study, the incidence of hypoxia of subjects in group EL, group EM and group EH (0, 2.3, 2.3%) was significantly lower compared with that of patients in group F (11.1%) (χ2 = 8.622; P = 0.035). The findings indicated that the incidence of somatic motor reactions was significantly lower in participants in group EM and group EH (9.1, 4.5%) relative to that of patients in group F and group EL (26.7, 15.6%) (χ2 = 10.254; P = 0.016). The results showed that the incidence of nausea and vomiting and potential psychiatric symptoms were significantly higher in patients in group EH (15.9, 11.4%) compared with that of participants in group F (2.2, 0%), group EL (4.4, 0%) and group EM (2.3, 2.3%) (χ2 = 7.493; P = 0.038 and χ2 = 8.248; P = 0.003). In this study, the mean arterial pressure (MAP) and heart rate (HR) of subjects in group EL, group EM and group EH were more stable compared with that of patients in group F. Frequency of the additional propofol dose was markedly less in group EM and EH (26.7%, 17,8%) compared with that in group F and EL (9.1, 4.5%) (χ2 = 10.254; P = 0.016). The findings indicated that the dischargeable time was significantly shorter for patients in group EM compared with that of subjects in group F, group EL and group EH. CONCLUSIONS: The findings of the present study showed that single-dose esketamine (0.25 mg kg- 1) effectively decreased incidence of hypotension and total adverse events and reduced the frequency of additional propofol dose required for patients undergoing painless abortion with preservation of physician-patient satisfaction.

Remimazolam tosilate in upper gastrointestinal endoscopy: A multicenter, randomized, non-inferiority, phase III trial.

BACKGROUND AND AIM: Remimazolam tosilate (RT) is a new short-acting GABA(A) receptor agonist, having potential to be an effective option for procedural sedation. Here, we aimed to compare the efficacy and safety of RT with propofol in patients undergoing upper gastrointestinal endoscopy. METHODS: This positive-controlled, non-inferiority, phase III trial recruited patients at 17 centers, between September 2017 and November 2017. A total of 384 patients scheduled to undergo upper gastrointestinal endoscopy were randomly assigned to receive RT or propofol. Primary endpoint was the success rate of sedation. Adverse events (AEs) were recorded to evaluate safety. RESULTS: The success rate of sedation in the RT group was non-inferior to that in the propofol group (97.34% vs 100.00%; difference in rate -2.66%, 95% CI -4.96 to -0.36, meeting criteria for non-inferiority). Patients in the RT group had longer time to adequate sedation (P < 0.0001) but shorter time to fully alert (P < 0.0001) than that in the propofol group. The incidences of hypotension (13.04% vs 42.86%, P < 0.0001), treatment-related hypotension (0.54% vs 5.82%, P < 0.0001), and respiratory depression (1.09% vs 6.88%, P = 0.0064) were significantly lower in the RT group. AEs were reported in 74 (39.15%) patients in the RT group and 114 (60.32%) patients in the propofol group, with significant difference (P < 0.0001). CONCLUSION: This trial established non-inferior sedation success rate of RT compared with propofol. RT allows faster recovery from sedation compared with propofol. The safety profile is favorable and appears to be superior to propofol, indicating that it was feasible and well tolerated for patients.

Long Noncoding RNA SNHG5 Knockdown Alleviates Neuropathic Pain by Targeting the miR-154-5p/CXCL13 Axis.

Neuropathic pain is an unneglectable pain condition with limited treatment options owing to its enigmatic underlying mechanisms. Long noncoding RNA small nucleolar RNA host gene 5 (SNHG5) is involved in the progression of a spectrum of human cancers. However, its role in neuropathic pain remains undiscovered. In the present study, we established a mouse spinal nerve ligation (SNL) model, and a significant upregulation of SNHG5 was observed. Then we knocked down SNHG5 level in mouse L5 dorsal root ganglion (DRG) by delivering specific short hairpin RNA against SNHG5 with adenovirus vehicle. Mouse paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) in response to mechanical stimuli was increased after SNHG5 knockdown, accompanied with decreased protein levels of glial fibrillary acidic protein (GFAP) and ionized calcium binding adapter molecule 1 (IBA-1). Besides, SNHG5 directly modulated the expression of miR-154-5p, which was downregulated in SNL mice. MiR-154-5p inhibition abolished the effect of SNHG5 knockdown on mouse behavioral tests and GFAP and IBA-1 levels. In addition, we validated that C-X-C motif chemokine 13 (CXCL13) was a novel downstream target of miR-154-5p, and CXCL13 level was positively related to that of SNHG5 in SNL mice. In conclusion, our study demonstrated that SNHG5 knockdown alleviated neuropathic pain and inhibited the activation of astrocytes and microglia by targeting the miR-154-5p/CXCL13 axis, which might be a novel therapeutic target for neuropathic treatment clinically.

Characterization and Fungicide Sensitivity of Colletotrichum Species Causing Strawberry Anthracnose in Eastern China.

Strawberry anthracnose caused by Colletotrichum spp. is one of the most serious diseases in the strawberry fields of China. In total, 196 isolates of Colletotrichum were obtained from leaves, stolons, and crowns of strawberry plants with anthracnose symptoms in eastern China and were characterized based on morphology, internal transcribed spacer (ITS), and ß-tubulin (TUB2) gene sequences. All 196 isolates were identified as the Colletotrichum gloeosporioides species complex. In total, 62 strains were further identified at the species level by phylogenetic analyses of multilocus sequences of ITS, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), Apn2-Mat1-2 intergenic spacer and partial mating type (ApMat), calmodulin (CAL), and TUB2. Three species from the C. gloeosporioides species complex were identified: Colletotrichum siamense, C. fructicola, and C. aenigma. Isolates of C. siamense were tolerant to high temperatures, with a significantly larger colony diameter than the other two species when grown above 36°C. The inoculation of strawberry plants confirmed the pathogenicity of all three species. C. siamense isolates resulted in the highest disease severity. The in vitro sensitivities of C. siamense and C. fructicola isolates to azoxystrobin and three demethylation-inhibitor (DMI) fungicides (difenoconazole, tebuconazole, and prochloraz) were determined. Both species were sensitive to DMI fungicides but not to azoxystrobin. C. siamense isolates were more sensitive to prochloraz, while C. fructicola isolates were more sensitive to difenoconazole and tebuconazole. The present study provides valuable information for the effective management of strawberry anthracnose.

Fast Quenching the Burst of Host Salicylic Acid Is Common in Early Strawberry/Colletotrichum fructicola Interaction.

The fungus Colletotrichum fructicola (a species of C. gloeosporioides complex) causes devastating anthracnose in strawberry. Like other species of the genus Colletotrichum, it uses a composite strategy including both the biotrophic and necrotrophic processes for pathogenesis. Host-derived hormones are central regulators of immunity, among which salicylic acid (SA) is the core defense one against biotrophic and hemibiotrophic pathogens. However, the manner and timing of pathogen manipulation of SA are largely elusive in strawberry. To achieve better understanding of the early challenges that SA-mediated defense experiences during strawberry/C. fructicola interaction, dynamic changes of SA levels were followed through the high-performance liquid chromatography method. A very early burst of free SA at 1 h postinoculation (hpi) followed by a fast quenching during the next 12 h was noticed, although rhythm variations were present in two hosts. Transcriptional characterization of genes related to SA pathway in two varieties on C. fructicola inoculation revealed that these genes were differentially expressed, although they were all induced at different time points. At the same time, three types of genes encoding homologous effectors interfering with SA accumulation were found to be first inhibited but sequentially activated during the first 24 hpi. Furthermore, subcellular localization analysis suggests that CfShy1 is a weapon of C. fructicola for strawberry invasion. Based on these results, we propose that the infection strategy that C. fructicola utilizes on strawberry is specialized, which is implemented through the optimized expression of a specific set of effector genes. Transcriptional characterization of host genes supports that de novo SA biosynthesis and the free SA release from methyl salicylate might contribute equally to the intricate control of SA homeostasis in strawberry. C. fructicola manipulation of SA-dependent resistance in strawberry might be closely related to multihormonal interplay among SA, jasmonic acid, abscisic acid, and cytokinin.

[Impact of inhaled anesthetics on the male reproductive system and its mechanisms: An update].

The safety of occupational exposure to inhaled anesthetics remains a concern among the medical staff in hospitals. Few reports are seen about the impact of inhaled anesthetics on the reproductive system, particularly that of males. Several clinical and basic studies on isoflurane and others suggest that inhaled anesthetics affect the reproductive system of rodents by decreasing the sperm count, inducing sperm morphological abnormality, reducing sperm motility, and changing the levels of reproductive hormones, the underlying mechanisms of which are mainly associated with the alteration of the hypothalamic-pituitary-gonadal axis and DNA damage and apoptosis of reproductive cells. This article reviews the main impacts of inhaled anesthetics on the male reproductive system and the possible mechanisms.

Vital Staining of Bacteria by Sunset Yellow Pigment.

In this study, we describe a method for discriminating pathogenic bacteria with a dye. First, we determined that among several colours tested, the sunset yellow pigment easily coloured Escherichia coli bacteria yellow. Next, we demonstrated that E. coli O157:H7, Shigella flexneri O301, Staphylococcus aureus and Bacillus subtilis could all be well marked by sunset yellow pigment. Finally, we performed bacterial viability assays and found there was no effect on bacterial growth when in co-culture with sunset yellow. Our results suggest that sunset yellow is suitable pigment to dye microorganisms.