Flotation separation of coal dust from foundry dust enhanced by pre-soaking assisted mechanical stirring.

Foundry dust is the main refractory solid waste in the foundry industry, and its resource utilization is a top priority for realizing green and cleaner production. The massive amount of coal dust in foundry dust is a potential impediment to the recycling of foundry dust, and the efficient separation of coal dust is crucial to solving the above problems. In this paper, the flotation separation of coal dust from foundry dust enhanced by pre-soaking assisted mechanical stirring was reported. The influence of pre-soaking, stirring speed, and stirring time on the flotation results of foundry dust was systematically studied, and the enhancement mechanism was analyzed based on the microstructure and hydrophobicity of foundry dust. Flotation kinetics experiments with different stirring time were conducted to clarify the flotation process of foundry dust. The results indicate that the pre-soaking of foundry dust is beneficial for the water-absorbing swelling of clay minerals coated on the surface of coal dust, and the subsequent mechanical stirring pretreatment promotes the monomer dissociation of foundry dust, which increases the contact angle of foundry dust and considerably improves the flotation results. The optimal stirring speed and stirring time were 2400 rpm and 30 min, respectively. The classical first-order model presented the highest degree of fitting with the flotation data among the five flotation kinetics models. Therefore, the pre-soaking assisted mechanical stirring is a promising method for promoting flotation separation and the complete recycling of foundry dust.

A novel ESIPT fluorescent probe derived from 3-hydroxyphthalimide for hydrazine detection in aqueous solution and living cells.

Hydrazine is a highly toxic and flammable liquid that can damage human liver, kidney, and central nervous system. Therefore, it is valuable to seek a quick and sensitive method for hydrazine detection in environmental and biological science. Herein, a new fluorescent probe derived from 3-hydroxyphthalimide was synthesized. This probe can rapidly and selectively detect hydrazine with a low detection limit of 4.3 × 10-7 M. The recognition principle is based on hydrazine-induced acetyl deprotection and excited-state intramolecular proton transfer (ESIPT) process. Moreover, test paper and fluorescence image experiments showed that this probe had potential to monitor hydrazine in the environment and living cells.

Coumarin 1,4-enedione for selective detection of hydrazine in aqueous solution and fluorescence imaging in living cells.

Hydrazine is a widely used but highly toxic chemical reagent, and the development of a fluorescent probe for hydrazine detection is very meaningful. In this study, a novel coumarin-derived fluorescent probe containing a 1,4-enedione moiety for hydrazine detection was developed. The recognition of hydrazine with the probe brings about obvious fluorescence enhancement over other environmentally relevant ions and amine-containing species. The limit of detection for hydrazine is 2.7×10-8 M in aqueous solution. The fluorescence enhancement was ascribed to the cyclization reaction of the 1,4-enedione moiety of the probe and hydrazine which form a six-membered pyridazine ring and intramolecular charge transfer (ICT) mechanism. The mass spectrometry (MS), nuclear magnetic resonance (NMR) analysis and theoretical calculations confirmed the recognition produced. The probe can be used to determine trace hydrazine in real water samples. More importantly, the probe also showed good potential in detecting hydrazine by imaging of living HeLa cells.

Traumatic prolapse of the globe into the anterior cranial fossa: a case report.

BACKGROUND: Orbital fracture associated with traumatic intracranial prolapse of the eyeball is rare. In all previously reported cases, vision was severely impaired with no light perception. Herein, we report a case of traumatic prolapse of the globe into the anterior cranial fossa, in which the patient's vision was preserved by early repositioning. CASE PRESENTATION: The present case report focused on a man hit by a steel pipe, leading to prolapse of the globe of the right eye into the anterior cranial fossa through fractures in the superior orbit roof, accompanied by cerebral contusion. The eyeball was immediately repositioned into the orbital cavity, along which the wound tract was debrided and the skull base was repaired. The patient underwent a follow-up period of 12 months, during which the visual acuity increased to 12/20 without any intracranial infections. However, the patient's ptosis persisted and was associated with complete loss of supraduction. CONCLUSIONS: In this case, early diagnosis and proper globe repositioning with reconstruction of the orbital roof could allow recovery of vision, as well as prevention of intracranial infection.

Methylated of genes behaving as potential biomarkers in evaluating malignant degree of glioblastoma.

Abnormal methylation genes usually act as oncogenes or anti-oncogenes in the occurrence and development of tumor, indicating their potential role as biomarkers in the evaluation of malignant tumor. However, the research on methylation's association with glioblastoma was rare. We attempted to figure out whether the methylation of genes could serve as the biomarker in evaluating the malignant degree of GBM. Methylation microarray data of 275 GBM patients have been downloaded from The Cancer Genome Atlas (TCGA) dataset. Logistic regression was used to find the methylated genes associated with the malignant degree of patients with the tumor. Functional enrichment analysis and network analysis were further performed on these selected genes. A total of 668, 412, 470, and 620 genes relevant with the methylation or demethylation were found to be associated with the malignant degree, Grades 1-4 of tumor. The higher the degree of malignant tumor, the higher of its methylation degree of its corresponding genes. GO and KEGG analysis results showed that these methylated genes were enriched in many functions as cell adhesion, abnormal transcription, and cell cycle disorder, etc. Of note, CCL11 and LCN11 were found to be significantly related to the progression of GBM. Critical genes associated with cell cycle as CCL11 and LCN1 may play essential roles in the occurrence, development, and transition of glioblastoma. More research was needed to explore its potential molecular mechanism.

New Colorimetric and Fluorometric Fluoride Ion Probe Based on Anthra[1,9-cd]pyrazol-6(2H)-one.

New colorimetric and fluorometric fluoride ion probe, anthra[1,9-cd]pyrazol- 6(2H)-one (1), was synthesized by one-step condensation. The probe 1 shows F--selective color change from colorless to pink and appearance of red fluorescence. The fluorescence quantum yield of free probe 1 in DMSO was calculated to be 0.03. After addition of 15 equiv. of F-, its fluorescence quantum yield can be increased to 0.37. The analytical detection limit for F- was 2.8 × 10- 7 M. 1H NMR analysis and DFT calculation show that the F--induced colorimetric and fluorometric responses of 1 are driven by deprotonation process. Graphical Abstract.

Comparative transcriptome analyses of deltamethrin-susceptible and -resistant Culex pipiens pallens by RNA-seq.

The widespread and improper use of pyrethroid insecticides, such as deltamethrin, has resulted in the evolution of resistance in many mosquito species, including Culex pipiens pallens. With the development of high-throughput sequencing, it is possible to massively screen pyrethroid resistance-associated gene. In this study, we used Illumina-Solexa transcriptome sequencing to identify genes that are expressed differently in deltamethrin-susceptible and -resistant strains of Culex pipiens pallens as a critical knowledge base for further studies. A total of 4,961,197,620 base pairs and 55,124,418 reads were sequenced, mapped to the Culex quinquefasciatus genome and assembled into 17,679 known genes. We recorded 1826 significantly differentially expressed genes (DEGs). Among them, 1078 genes were up-regulated and 748 genes were down-regulated in the deltamethrin-resistant strain compared to -susceptible strain. These DEGs contained cytochrome P450 s, cuticle proteins, UDP-glucuronosyltransferases, lipases, serine proteases, heat shock proteins, esterases and others. Among the 1826 DEGs, we found that the transcriptional levels of CYP6AA9 in the laboratory populations was elevated as the levels of deltamethrin resistance increased. Moreover, the expression levels of the CYP6AA9 were significantly higher in the resistant strains than the susceptible strains in three different field populations. We further confirmed the association between the CYP6AA9 gene and deltamethrin resistance in mosquitoes by RNA interfering (RNAi). Altogether, we explored massive potential pyrethroid resistance-associated genes and demonstrated that CYP6AA9 participated in the pyrethroid resistance in mosquitoes.

MiR-278-3p regulates pyrethroid resistance in Culex pipiens pallens.

MicroRNAs (miRNAs) regulate gene expression and biological processes including embryonic development, innate immunity, and infection in many species. Emerging evidence indicates that miRNAs are involved in drug resistance. However, little is known about the relationship between the miRNAs and insecticide resistance in mosquitos. Here, we reported that conserved miR-278-3p and its target gene are critical for pyrethroid resistance in Culex pipiens pallens. We found that CYP6AG11 is the target of miR-278-3p, through bioinformatic analysis and experimental verification. The expression level of miR-278-3p was lower, whereas the level of CYP6AG11 was higher in deltamethrin-resistant strain, which were detected using quantitative reverse transcription PCR (qRT-PCR). We also found that CYP6AG11 was regulated by miR-278-3p via a specific target site with the 3' untranslated region (UTR) by luciferase reporter assay. In addition, overexpression of CYP6AG11 in the mosquito C6/36 cells showed better proliferation than the cells with empty vector when treated by deltamethrin at different concentrations. Moreover, the overexpression of miR-278-3p through microinjection led to a significant reduction in the survival rate, and the level of CYP6AG11 was simultaneously reduced. These results indicated that miR-278-3p could regulate the pyrethroid resistance through CYP6AG11.

Imbalance of Ca2+ and K+ fluxes in C6 glioma cells after PDT measured with scanning ion-selective electrode technique.

Photodynamic therapy (PDT) possesses the capacity to lead to death of C6 glioma in vitro and in vivo. The purpose of this study was to investigate whether Ca(2+) and K(+) homeostasis of C6 glioma cells were affected by PDT. C6 glioma cells were randomly divided into five groups: control group, Hematoporphyrin derivative (HpD) group (10 mg/l, without irradiation), PDT group (HpD 10 mg/l + irradiation), PDT&6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX) group (HpD 10 mg/l + CNQX 50 mol/l + irradiation), and HpD&CNQX group (HpD 10 mg/l + CNQX 50 mol/l, without irradiation). Glioma cells in PDT and PDT&CNQX group were subjected to PDT. Cells in PDT&CNQX group were administered α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptor antagonist CNQX prior to PDT on C6 glioma cells. The changes of Ca(2+) and K(+) fluxes were studied by using a non-invasive scanning ion-selective electrode technique (SIET). Morphology of C6 cells was observed with optical microscopy. PDT induced Ca(2+) influx and K(+) efflux significantly, which resulted in death of C6 cells. When AMPA glutamate receptor antagonist CNQX was applied, Ca(2+) influx and K(+) efflux were partly blocked up and viability of C6 cells increased. These results indicate that Ca(2+) influx and K(+) efflux may correlate with the treatment effects of PDT on C6 glioma cells.

Clot removAl with or without decompRessive craniectomy under ICP monitoring for supratentorial IntraCerebral Hemorrhage (CARICH): a randomized controlled trial.

BACKGROUND: Decompressive craniectomy (DC), a surgery to remove part of the skull and open the dura mater, maybe an effective treatment for controlling intracranial hypertension. It remains great interest to elucidate whether DC is beneficial to intracerebral hemorrhage (ICH) patients who warrant clot removal (CR) to prevent intracranial hypertension. METHODS: The trial was a prospective, pragmatic, controlled trial involving adult patients with ICH who were undergoing removal of hematoma. ICH patients were randomly assigned at a 1:1 ratioto undergo CR with or without DC under the monitoring of intracranial pressure. The primary outcome was the proportion of unfavorable functional outcome (modified Rankin Scale 3-6) at 3 months. Secondary outcomes included the mortality at 3 months and the occurrence of reoperation. RESULTS: A total of 102 patients were assigned to the CR with DC group and 102 to the CR group. Median hematoma volume was 54.0 ml (range 30-80 ml) and median preoperative Glasgow Coma Scale was 10 (range 5-15). At 3 months, 94 patients (92.2%) in CR with DC group and 83 patients (81.4%) in the CR group had unfavorable functional outcome ( P =0.023). Fourteen patients (13.7%) in the CR with DC group died versus five patients (4.9%) in the CR group ( P =0.030). The number of patients with reoperation was similar between the CR with DC group and CR group (5.9 vs. 3.9%; P =0.517). Postoperative intracranial pressure values were not significantly different between two groups and the mean values were less than 20 mmHg. CONCLUSIONS: CR without DC decreased the rate of modified Rankin Scale score of 3-6 and mortality in patients with ICH, compared with CR with DC.

Amelioration of White Matter Injury Through Mitigating Ferroptosis Following Hepcidin Treatment After Spinal Cord Injury.

Spinal cord injury (SCI) usually introduces permanent or long-lasting neurological impairments. Maintaining the integrity of the limited number of white matter bundles (5-10%) preserves wholly or partially locomotor following SCI. Considering that the basic structure of white matter bundles is axon wrapped by oligodendrocytes, promoting oligodendrocytes survival might be a feasible strategy for reducing white matter injury (WMI) after SCI. Oligodendrocytes are rich in unsaturated fatty acid and susceptible to ferroptosis-induced damage. Hence, exploring method to reduce ferroptosis is supposed to expedite oligodendrocytes survival, thereafter mitigating WMI to facilitate functional recovery post-SCI. Here, the results indicated the administration of hepcidin reduced iron accumulation to promote oligodendrocytes survival and to decrease spinal cord atrophy, therefore facilitating functional recovery. Then, the WMI was evidently decreased owing to attenuating ferroptosis. Subsequently, the results revealed that the expression of divalent metal transporter 1 (DMT1) and transferrin receptor (TfR) was expressed in CC1+ cells. The expression level of DMT1 and TfR was significantly increased, while this phenomenon was obviously neutralized with the administration of hepcidin in the epicenter of spinal cord after SCI. Afterward, the application of hepcidin downregulated reactive oxygen species (ROS) overload, which was evidently increased with the treatment of 20 µM FeCl3, therefore increasing cell viability and reducing lactate dehydrogenase (LDH) activity through downregulating the expression of DMT1 and TfR to inhibit ferroptosis in oligodendrocyte progenitor cells (OPCs). The present study provides evidence that the application of hepcidin facilitates oligodendrocytes survival to alleviate WMI via reducing the expression of DMT1 and TfR.

Exosomes from young healthy human plasma promote functional recovery from intracerebral hemorrhage via counteracting ferroptotic injury.

Intracerebral hemorrhage (ICH), as a type of life-threatening and highly disabled disease, has limited therapeutic approaches. Here, we show that exosomes derived from young healthy human plasma exhibiting typical exosomes features could facilitate functional recovery of ICH mice. When these exosomes are intraventricularly delivered into the brain after ICH, they mainly distribute around the hematoma and could be internalized by neuronal cells. Strikingly, exosomes administration markedly enhanced the behavioral recovery of ICH mice through reducing brain injury and cell ferroptosis. MiRNA sequencing revealed that microRNA-25-3p (miR-25-3p) was differentially expressed miRNA in the exosomes from young healthy human plasma, compared with exosomes from the old control. Importantly, miR-25-3p mimicked the treatment effect of exosomes on behavioral improvement, and mediated the neuroprotective effect of exosomes against ferroptosis in ICH. Furthermore, luciferase assay and western blotting data illustrated that P53 as assumed the role of a downstream effector of miR-25-3p, thereby regulating SLC7A11/GPX4 pathway to counteract ferroptosis. Taken together, these findings firstly reveal that exosomes from young healthy human plasma improve functional recovery through counteracting ferroptotic injury by regulating P53/SLC7A11/GPX4 axis after ICH. Given the easy availability of plasma exosomes, our study provides a potent therapeutic strategy for ICH patients with quick clinical translation in the near future.

In vivo magnetic resonance imaging tracking of SPIO-labeled human umbilical cord mesenchymal stem cells.

Human umbilical cord mesenchymal stem cells (hUC-MSCs) can be efficiently labeled by superparamagnetic iron oxide (SPIO) nanoparticles, which produces low signal intensity on magnetic resonance imaging (MRI) in vitro. This study was to evaluate the feasibility of in vivo tracking for hUC-MSCs labeled by SPIO with noninvasive MRI. SPIO was added to cultures at concentrations equivalent to 0, 7, 14, 28, and 56 µg Fe/ml (diluted with DMEM/F12) and incubated for 16 h. Prussian Blue staining was used to determinate the labeling efficiency. Rats were randomly divided into three groups, control group, hUC-MSCs group, and SPIO-labeled hUC-MSCs group. All groups were subjected to spinal cord injury (SCI) by weight drop device. Rats were examined for neurological function. In vivo MRI was used to track SPIO-labeled hUC-MSCs transplanted in rats spinal cord. Survival and migration of hUC-MSCs were also explored using immunofluorescence. Significant improvements in locomotion were observed in the hUC-MSCs groups. There was statistical significance compared with control group. In vivo MRI 1 and 3 weeks after injection showed a large reduction in signal intensity in the region transplanted with SPIO-labeled hUC-MSCs. The images from unlabeled hUC-MSCs showed a smaller reduction in signal intensity. Transplanted hUC-MSCs engrafted within the injured rats spinal cord and survived for at least 8 weeks. In conclusion, hUC-MSCs can survive and migrate in the host spinal cord after transplantation, which promote functional recovery after SCI. Noninvasive imaging of transplanted SPIO-labeled hUC-MSCs is feasible.

G-protein coupled estrogen receptor 1 mediated estrogenic neuroprotection against spinal cord injury.

OBJECTIVE: What underlies the protection of estrogen against spinal cord injury remains largely unclear. Here, we investigated the expression pattern of a new estrogen receptor, G-protein coupled estrogen receptor 1 in the spinal cord and its role in estrogenic protection against spinal cord injury. DESIGN AND SETTINGS: Department of Neurosurgery and Key Laboratory of Neurotrauma, Southwest Hospital. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: The animals subjected to spinal cord injury were divided into six groups and given vehicle solution, 17ß-estradiol, or G-protein coupled estrogen receptor 1 agonist G-1 at 15 mins and 24 hrs postinjury, or given nuclear estrogen receptor antagonist ICI 182,780 at 1 hr before spinal cord injury followed by 17ß-estradiol administration at 15 mins and 24 hrs postinjury, or given G-protein coupled estrogen receptor 1 specific antisense or random control oligonucleotide at 4 days before spinal cord injury followed by 17ß-estradiol administration at 15 mins and 24 hrs postinjury. MEASUREMENTS: Male Sprague-Dawley rats were subjected to spinal cord injury using a weight-drop injury approach. Immunohistochemical assays were used to observe the distribution and cell-type expression pattern of G-protein coupled estrogen receptor 1. The terminal deoxynucleotidyl transferase dUTP nick-end labeling-staining assay and behavior tests were employed to assess the role of G-protein coupled estrogen receptor 1 in mediating estrogenic protection against spinal cord injury. MAIN RESULTS: We show that G-protein coupled estrogen receptor 1 is mainly distributed in the ventral horn and white matter of the spinal cord, which is totally different from nuclear estrogen receptors. We also show that G-protein coupled estrogen receptor 1 is specifically expressed by neurons, oligodendrocytes, and microglial cells, but not astrocytes. Furthermore, estrogen treatment prevents spinal cord injury-induced apoptotic cell death and enhances functional recovery after spinal cord injury, which can be mimicked by the specific G-protein coupled estrogen receptor 1 agonist G-1 and inhibited by specific knockdown of G-protein coupled estrogen receptor 1 expression, but not pure nuclear ER antagonist ICI 182,780. Finally, we show that estrogen or G-1 up-regulates the protein expression level of G-protein coupled estrogen receptor 1 to intensify estrogenic effects during spinal cord injury. CONCLUSIONS: These results reveal that G-protein coupled estrogen receptor 1 may mediate estrogenic neuroprotection against spinal cord injury, and underline the promising potential of estrogen with its new target G-protein coupled estrogen receptor 1 for the treatment of spinal cord injury patients.

Ferrostatin-1 Alleviates White Matter Injury Via Decreasing Ferroptosis Following Spinal Cord Injury.

Spinal cord injury (SCI), a devastating neurological impairment, usually imposes a long-term psychological stress and high socioeconomic burden for the sufferers and their family. Recent researchers have paid arousing attention to white matter injury and the underlying mechanism following SCI. Ferroptosis has been revealed to be associated with diverse diseases including stroke, cancer, and kidney degeneration. Ferrostatin-1, a potent inhibitor of ferroptosis, has been illustrated to curb ferroptosis in neurons, subsequently improving functional recovery after traumatic brain injury (TBI) and SCI. However, the role of ferroptosis in white matter injury and the therapeutic effect of ferrostatin-1 on SCI are still unknown. Here, our results indicated that ferroptosis played a pivotal role in the secondary white matter injury, and ferrostatin-1 could reduce iron and reactive oxygen species (ROS) accumulation and downregulate the ferroptosis-related genes and its products of IREB2 and PTGS2 to further inhibit ferroptosis in oligodendrocyte, finally reducing white matter injury and promoting functional recovery following SCI in rats. Meanwhile, the results demonstrated that ferrostatin-1 held the potential of inhibiting the activation of reactive astrocyte and microglia. Mechanically, the present study deciphers the potential mechanism of white matter damage, which enlarges the therapeutic effects of ferrostatin-1 on SCI and even in other central nervous system (CNS) diseases existing ferroptosis.

Comprehensive utilization of foundry dust: Coal powder and clay minerals separation by ultrasonic-assisted flotation.

Clay sand casting generates a large amount of foundry dust (FD), and the presence of coal powder in the FD makes it difficult to recycle and utilize. The landfill of the FD creates a serious environmental pollution and wastes a valuable resource. To improve the above situation, the FD was analyzed and characterized by X-ray fluorescence spectrometer (XRF), X-ray diffraction (XRD) and electron probe microanalyzer (EPMA). An ultrasonic-assisted flotation process was developed for the comprehensive utilization of the FD, and the effects of ultrasonic time on the flotation performance and flotation kinetics were investigated. In addition, the two-stage flotation of the FD was conducted. Obtained results showed that the FD mainly consisted of coal powder and clay minerals, and the coal powder was covered by clay minerals. The separation efficiency of the coal powder and clay minerals can be significantly enhanced by ultrasonic pretreatment, and the optimal ultrasonic time was 30 min. The flotation kinetics analysis results indicated that the first-order model with rectangular distribution was more reasonable for the data fitting of the ultrasonic-assisted flotation. Furthermore, the concentrate and tailings obtained by the two-stage flotation had achieved an acceptable result, favoring the comprehensive utilization of the FD.

[Clinical application of self-made minimally invasive hood-assisted transforaminal lumbar interbody fusion via modified bilateral Wiltse approach in the treatment of lumbar degenerative diseases].

OBJECTIVE: To explore the advantages of self made minimally invasive hook assisted transforaminal lumbar interbody fusion (TLIF) via modified bilateral Wiltse approach in the treatment of lumbar degenerative diseases. METHODS: The clinical data of 140 patients underwent lumbar spine fusion surgery from October 2016 to October 2017 were retrospectively analyzed. Among them, 72 cases were treated by self-made minimally invasive hook-assisted TLIF via modified bilateral Wiltse approach (group A), there were 37 males and 35 females, aged (48±16) years old;68 cases were treated by TLIF via traditional posterior median approach (group B ), there were 38 males and 30 females, aged (45±15) years old. The surgical incision size, operation time, intraoperative blood loss volume, postoperative drainage volume, postoperative wound healing, and intervertebral fusion rate at the final follow-up were recorded between two groups. Visual analogue scale (VAS) and Oswestry Disability Index (ODI) were used to assess the clinical efficacy. RESULTS: All the patients were followed up for 3 to 13 (8±5) months. The wound in group A healed well after operation, and 1 case in group B occurred wound necrosis after operation, and healed after debridement and suture. There were no significant differences in operation time and postoperative fusion rate between two surgical methods (P>0.05). Group A had obvious advantages in surgical incision size, intraoperative blood loss volume and postoperative drainage volume (P<0.05), and the postoperative VAS score of low back pain and ODI were better than group B (P<0.05). CONCLUSION: The self made minimally invasive hook assistedTLIF via modified bilateral Wiltse approach has the characteristics of minimally invasive, less intraoperative blood loss, less postoperative drainage, fewer complications, and more stable fusion in the treatment of lumbar degenerative desease.

MiR-4448 is involved in deltamethrin resistance by targeting CYP4H31 in Culex pipiens pallens.

BACKGROUND: Culex pipiens (Cx. pipiens) complex, which acts as a vector of viruses and is widespread and abundant worldwide, including West Nile virus, Japanese encephalitis virus, and Sindbis virus, can cause serious vector-borne diseases affecting human health. Unfortunately, mosquitoes have developed deltamethrin resistance because of its long-term overuse, representing a major challenge to mosquito control. Understanding the molecular regulatory mechanisms of resistance is vital to control mosquitoes. MicroRNAs (miRNAs) are short non-coding RNAs that have been demonstrated to be important regulators of gene expression across a wide variety of organisms, which might function in mosquito deltamethrin resistance. In the present study, we aimed to investigate the regulatory functions of miR-4448 and CYP4H31 in the formation of insecticidal resistance in mosquito Culex pipiens pallens. METHODS: We used quantitative real-time reverse transcription PCR to measure miR-4448 and CYP4H31 (encoding a cytochrome P450) expression levels. The regulatory functions of miR-4448 and CYP4H31 were assessed using dual-luciferase reporter assays. Then, oral feeding, RNA interference, and the American Centers for Disease Control and Prevention bottle bioassay were used to determine miR-4448's association with deltamethrin resistance by targeting CYP4H31 in vivo. Cell Counting Kit-8 (CCK-8) was also used to detect the viability of pIB/V5-His-CYP4H31-transfected C6/36 cells after deltamethrin treatment in vitro. RESULTS: MiR-4448 was downregulated in the deltamethrin-resistant strain (DR strain), whereas CYP4H31 was downregulated in deltamethrin-susceptible strain. CYP4H31 expression was downregulated by miR-4448 recognizing and binding to its 3' untranslated region. Functional verification experiments showed that miR-4448 overexpression resulted in lower expression of CYP4H31. The mortality of miR-4448 mimic-injected DR strain mosquitoes was higher than that of the controls. CCK-8 assays showed that CYP4H31 decreased cellular resistance to deltamethrin in vitro and the mortality of the DR strain increased when CYP4H31 was knocked down in vivo. CONCLUSIONS: In mosquitoes, miR-4448 participates in deltamethrin resistance by targeting CYP4H31. The results of the present study increase our understanding of deltamethrin resistance mechanisms.

MiR-279-3p regulates deltamethrin resistance through CYP325BB1 in Culex pipiens pallens.

BACKGROUND: The overuse of insecticides to control insect vectors has promoted extensive insecticide resistance in mosquitoes. In this study, the functions of microRNA (miR)-279-3p and its target CYP325BB1 in the regulation of deltamethrin resistance in Culex pipiens pallens was investigated. METHODS: Quantitative real-time reverse transcription PCR was used to detect the expression levels of miR-279-3p and CYP325BB1. Then, the dual-luciferase reporter assay system, RNA interference, CDC bottle bioassay and Cell Counting Kit-8 (CCK-8) assay were used to explore the roles of these molecules in deltamethrin resistance both in vivo and in vitro. RESULTS: The expression patterns of miR-279-3p and CYP325BB1 were compared between deltamethrin-sensitive (DS-strain) and deltamethrin-resistant (DR-strain) mosquitoes. Luciferase activity was downregulated by miR-279-3p, the effect of which was ablated by a mutation of the putative binding site for CYP325BB1. In DR-strain mosquitoes, the expression of miR-279-3p was increased by microinjection and oral feeding of miR-279-3p agomir (mimic). CYP325BB1 mRNA levels were downregulated, which resulted in a higher mortality of the mosquitoes in miR-279-3p mimic-treated groups. In the DS-strain mosquitoes, microinjection of a miR-279-3p inhibitor decreased miR-279-3p expression, whereas the expression of CYP325BB1 was increased; the mortality of these mosquitoes decreased significantly. In addition, overexpression of pIB/V5-His-CYP325BB1 changed the sensitivity of C6/36 cells to deltamethrin in vitro. Also in DR-strain mosquitoes, downregulation of CYP325BB1 expression by microinjection of si-CYP325BB1 increased mosquito mortality in vivo. CONCLUSIONS: These findings provide empirical evidence of the involvement of miRNAs in the regulation of insecticide resistance and indicate that miR-279-3p suppresses the expression of CYP325BB1, which in turn decreases deltamethrin resistance, resulting in increased mosquito mortality. Taken together, the results provide important information for use in the development of future mosquito control strategies.

Combination of the Distance From Tumor Edge to Subventricular Zone and IDH Mutation Predicts Prognosis of Patients With Glioma.

Both subventricular zone (SVZ) contact and isocitrate dehydrogenase 1 (IDH1) mutation have been reported to be related to the outcome of glioma, respectively. However, far too little attention has been paid to the role of tumor edge-SVZ distance in the outcome of glioma. We aim to assess the value of tumor-SVZ distance, as well as combined tumor-SVZ distance and IDH status, in predicting the outcome of gliomas (WHO grade II-IV). Here, the MR images and clinical data from 146 patients were included in the current study. The relationship between survival and the tumor-SVZ distance as well as survival and combination of tumor-SVZ distance and IDH status were determined via univariate and multivariate analyses. In univariate analysis of tumor-SVZ distance, the patients were divided into three types (SVZ involvement, tumor-SVZ distance from 0 to 10 mm, and tumor-SVZ distance >10 mm). The results showed that the OS (p = 0.02) and PFS (p = 0.002) for the patients had a positive correlation with the tumor-SVZ distance. In addition, simple linear correlation found a significant relationship between the two parameters (OS and PFS) and tumor-SVZ distance in patients with non-SVZ-contacting glioma. Combination analysis of the tumor-SVZ distance and IDH status showed that IDH1 mutation and SVZ non-involvement enable favorable outcomes, whereas IDH1 wild type with SVZ involvement indicates a significantly worse prognosis in all patients. Moreover, in patients with non-SVZ-contacting glioma, IDH1 mutation concurrent with tumor-SVZ distance >10 mm has better OS and PFS. IDH1 wild type and tumor-SVZ distance from 0 to 10 mm suggest poorer OS and PFS. Multivariate analysis showed WHO grade IV, SVZ involvement, tumor-SVZ distance from 0 to 10 mm, IDH1 mutation, gross total resection, and chemotherapy serve as independent predictors of OS. WHO grade IV, SVZ involvement, tumor-SVZ distance from 0 to 10 mm, IDH1 mutation, and chemotherapy serve as independent predictors of PFS of patients with glioma. In conclusion, tumor-SVZ distance and IDH1 mutation status are the determinants affecting patient outcome.