Integrated Microwell Array-Based Microfluidic Chip with a Hand-Held Smartphone-Controlled Device for Nucleic Acid Detection.

In this study, we designed a highly integrated microfluidic chip for nucleic acid extraction, amplification, and detection. Magnetic beads, which are used to capture nucleic acids on the chip, are trapped in the microwell arrays in a one-well-one-bead manner after local surface modification of the inner faces of the microwells. On-chip liquid introduction, delivery, and mixing are all carried out manually with one syringe and no other equipment. A hand-held device with precise temperature control and high-quality imaging is developed, which is only 2.3 cubic decimeters in volume and 1.2 kg in weight. Via the use of the Internet for wireless communication, the experiment and data analysis after inserting the chip into the device can be conducted by a smartphone anywhere there is an Internet connection. We carried out reverse transcription loop-mediated isothermal amplification (RT-LAMP) on the chip with the hand-held device. SARS-CoV-2 pseudoviruses are extracted, reverse transcribed, amplified, and detected on the chip with the hand-held device with satisfactory results. Thus, a highly integrated, easy-to-operate, and rapid nucleic acid detection microfluidic chip with a hand-held smartphone-controlled device is proposed, and this new platform for nucleic acid detection shows great potential for mobile point-of-care testing (POCT).

Effect of music listening on perioperative anxiety, acute pain and pain catastrophizing in women undergoing elective cesarean delivery: a randomized controlled trial.

BACKGROUND: Anxiety may adversely impact mother and her newborn. Music listening is a safe and efficacious treatment that may to reduce perioperative anxiety. The effect on acute pain and pain catastrophizing scores remains unclear. We aimed to determine whether perioperative music listening reduces anxiety, acute pain, and pain catastrophizing scale (PCS) scores following elective cesarean delivery under spinal anesthesia. METHODS: After randomization into music listening and control groups, baseline patient characteristics, visual analog scale-anxiety (VAS-A) scores, pain scores, PCS total and sub-scores, and music preferences were collected preoperatively. Before surgery, parturients in the experimental group listened to music of their own choice for 30 min. Music listening was continued during administration of spinal anesthesia and cesarean delivery, and for 30 min following surgery. Postoperative VAS-A score, acute pain score, PCS scores, music preferences, satisfaction score, and feedback were recorded. RESULTS: We analyzed 108 parturients (music: n = 53; control: n = 55). Music listening was associated with reduced postoperative VAS-A (mean difference (MD) -1.43, 95%CI -0.63 to -2.22), PCS total score (MD -6.39, 95%CI -2.11 to -10.66), PCS sub-scores on rumination (MD -1.68, 95%CI -0.12 to -3.25), magnification (MD -1.53, 95%CI -0.45 to -2.62), and helplessness (MD -3.17, 95%CI -1.29 to -5.06) sub-scores. There was no significant difference in postoperative acute pain scores. The majority (> 95%) of parturients reported "excellent" and "good" satisfaction with music listening, and most provided positive feedback. CONCLUSION: Perioperative music listening was associated with reduced postoperative anxiety and lower pain catastrophizing. Based on the good patient satisfaction and positive feedback received, the use of music listening in the obstetric setting is recommended. TRIAL REGISTRATION: This study was registered on Clinicaltrials.gov NCT03415620 on 30/01/2018.

Interleukin-1 receptor associated kinase (IRAK)-M -mediated type 2 microglia polarization ameliorates the severity of experimental autoimmune encephalomyelitis (EAE).

Toll-like receptor 4 (TLR4) play a key role in activating the innate immune system during pathogen recognition. In the pathogenesis of multiple sclerosis (MS), activated TLR4 together with myeloid differentiation primary response gene 88 (MyD88) produce an inflammatory microenvironment that promotes the differentiation of microglia into the M1 phenotype, who plays a key role in the pathogenesis of MS. Interleukin-1 receptor-associated kinase (IRAK)-M is specifically expressed in microglia in central nervous system (CNS) and act as a negative regulator of TLR4-MyD88 signaling pathway. Moreover, previous studies have shown that IRAK-M promotes the differentiation of type 2 microglia; however, its role in MS has not been explored. In the present study, we demonstrated that IRAK-M expression is elevated during EAE, and IRAK-M-/- mice significantly accelerated course and increased severity of disease, accompanied by a visible increase of the M1 microglia infiltrated. In conclusion, these data indicates that IRAK-M significantly improves EAE onset through down-regulation of the TLR4-MyD88 signaling pathway, which finally leads to differentiation of M2 phenotype in the microglia. Our study suggests that IRAK-M may be a potential therapeutic target for the treatment of MS.

Straightforward and Ultrastable Surface Modification of Microfluidic Chips with Norepinephrine Bitartrate Improves Performance in Immunoassays.

Polymers are commonly used materials for microfluidic chip fabrication, because they are standardized in fabrication and low in cost. However, most polymeric materials that are readily fabricated on the industrial scale are hydrophobic, which is inconvenient for the injection and flow of the aqueous solution, resulting in poor analytical performance for biochemical assays. In this work, we present a straightforward and ultrastable surface modification process for polymeric chips. A one-step modification by using norepinephrine bitartrate monohydrate as a modification reagent is completed at room temperature. The hydrophilicity of the polymeric surfaces increases dramatically. Surface modification is stable for at least 2.5 years, allowing for autoinjection of aqueous solution into the channels. The chips are applied in the immunoassay of alpha-fetoprotein (AFP). The low nonspecific adsorption after modification results in significantly decreased background noise, optimized signal-to-noise ratios (SNR), and dramatically enhanced reproducibility of the immunoassay. Thirty clinical human serum samples are analyzed; these results strongly correlated with the values obtained using commercial test kits. We anticipate that this surface modification method can be used for immunoassay devices in analytical and biosensing technology.

Capillary-based chemiluminescence immunoassay for C-reactive protein with portable imaging device.

A capillary-based chemiluminescence immunoassay system using a charge-coupled device (CCD) camera as detector was established in this paper. The fused quartz capillary was easily activated in one step for immobilizing capture antibody, and the chemiluminescence immunoassay was carried out in the capillary in double-antibody sandwich format. Chemiluminescence signals were recorded by the portable imaging device which was installed with the CCD camera and the results were analyzed through gray intensity. The total cost time, which included not only the time for test but also the time for the preparation of experimental materials, was only 2 h. The immunoassay was performed without any complicated or expensive instruments. The consumption of the sample was only 0.8 µL in one test, which was significantly less than other methods. In this work, C-reactive protein (CRP), as a target, was quantitatively detected from 0.3 to 160.0 µg mL-1 with high specificity and low sample volume. The reproducibility and accuracy were tested in clinic human serum samples and shown good results. Thus, this rapid, easy preparation and using, portable immunoassay system indicated its usefulness as a novel technology platform. Graphical abstract ᅟ.

Chronic mountain sickness in Chinese Han males who migrated to the Qinghai-Tibetan plateau: application and evaluation of diagnostic criteria for chronic mountain sickness.

BACKGROUND: Chronic mountain sickness (CMS), originally characterized by excess hemoglobin (Hb), is currently diagnosed using score-based diagnostic criteria combined with excessive erythrocytosis and clinical symptoms. However, the current criteria have limited applicability. We applied these criteria to 1,029 Chinese Han males migrated to and have been stayed at the Qinghai-Tibet plateau (3,700-5,000 m) for 2-96 months to investigate the prevalence of CMS and its correlations with Hb concentration, altitude, and the length of residence. METHODS: Subjects were screened for CMS using the latest approved diagnostic criteria combined with excessive erythrocytosis and clinical symptoms. Hb concentrations were measured, and a cut-off point was determined with k-means clustering. Predisposing factors were evaluated with binary logistic analysis and curve fitting analysis. RESULTS: (1) The prevalence of CMS at the Qinghai-Tibetan plateau was 17.8% (183/1029 subjects, with CMS score ≥ 6, and Hb ≥ 210 g/L), which is higher than that previously reported. (2) While individuals were identified into two Hb clusters with a cut-off point of 200 g/L, in the low-Hb cluster (Hb < 200 g/L), the oxygen saturation remained stable as the Hb increased; in the high-Hb cluster (Hb ≥ 200 g/L), the oxygen saturation decreased as the Hb increased. (3) Two critical factors associated with CMS development were residence at an altitude of 4,500 m and a 60-month length of residence. CONCLUSIONS: Our presenting scoring system is more sensitive than previous diagnostic criteria and favors early screening and treatment of patients with CMS. Our finding suggests that an adjusted Hb threshold of 200 g/L (instead of 210 g/L) is more adaptable in Han individuals at all altitudes. The weight of Hb level should score ≥ 6 points using the CMS scoring system because of the pathophysiologic role of excessive erythrocytosis in patients with CMS. In addition, our data suggest the importance of early screening of CMS via regular medical examinations within the first 60 months of residence at high altitudes, especially >4500 m.

[Effects of adenosine on hMLH1 methylation of human colorectal cancer cells].

OBJECTIVE: To explore the effects of adenosine on hMLH1 methylation of human colorectal cancer cells. METHODS: The SW480 cells were treated with adenosine at the concentrations of 0, 1.5, 3.0, 4.5 mmol/L for 72 h. The hMLH1 methylation levels of CpG islands were detected by bisulfite sequencing polymerase chain reaction (BSP), hMLH1 mRNA expression levels by reverse transcription-polymerase chain reaction (RT-PCR), the expression levels of hMLH1 protein by Western blot and the apoptotic rates by flow cytometry (FCM). The cells were treated with adenosine at the concentrations of 0, 1.5, 3.0, 4.5 mmol/L for 24, 48, 72, 96 h. And their proliferation rates were detected by methyl thiazolyl tetrazolium (MTT). RESULTS: After a 72 h treatment of adenosine, the hMLH1 promoter methylation levels of 1.5, 3.0 and 4.5 mmol/L groups were 65% ± 4%, 45% ± 11% and 16% ± 4% respectively and were all significantly lower than that of the control group (80% ± 4%, all P < 0.01). The mRNA expression levels, hMLH1 protein expression levels and apoptotic rates were all significantly higher than that of the control group (0.230 ± 0.032, 0.359 ± 0.029 and 0.570 ± 0.019 vs 0.079 ± 0.010; 0.353 ± 0.016, 0.654 ± 0.018 and 0.854 ± 0.014 vs 0.126 ± 0.016; 11.9% ± 0.6%, 20.0% ± 1.8% and 35.8% ± 1.8% vs 3.9% ± 1.4%, all P < 0.01). MTT showed that the proliferation rates of SW480 cells were lower than that of the control group and a time-dosage dependence existed (all P < 0.05). CONCLUSION: Adenosine can reverse the abnormal methylation of hMLH1 CpG island and promote the expression of hMLH1 so as to restrain the proliferation and promote the apoptosis of colocectal cancer cells.

EsxA might as a virulence factor induce antibodies in patients with Staphylococcus aureus infection.

Staphylococcus aureus (S. aureus) is an important human pathogen, which commonly causes the acquired infectious diseases in the hospital and community. Effective and simple antibiotic treatment against S. aureus-related disease becomes increasingly difficult. Developing a safe and effective vaccine against S. aureus has become one of the world's hot spots once again. The key issue of developing the vaccine of S. aureus is how to find an ideal key pathogenic gene of S. aureus. It was previously suggested that EsxA might be a very important factor in S. aureus abscess formation in mice, but clinical experimental evidence was lacking. We therefore expressed EsxA protein through prokaryotic expression system and purified EsxA protein by Ni-affinity chromatography. ELISA was used to detect the anti-EsxA antibodies in sera of 78 patients with S. aureus infection and results showed that the anti-EsxA antibodies were positive in the sera of 19 patients. We further analyzed the EsxA positive antibodies related strains by antimicrobial susceptibility assay and found that all of the corresponding strains were multi-drug resistant. Among those multi-drug resistant strains, 73.7% were resistant to MRSA. The results indicated EsxA is very important in the pathogenesis of S. aureus. We suggested that the EsxA is very valuable as vaccine candidate target antigens for prevention and control of S. aureus infection.

Microfluidic on-chip valve and pump for applications in immunoassays.

On-chip valves can simplify a microfluidic chip and make it easy to operate. However, most on-chip valves already reported still need complicated manufacture and sophisticated supporting devices. In this work, we present a straightforward on-chip valve, which can be serially connected, to form an on-chip pump. The liquid can horizontally flow one way by the regular deformations of flexure strips in the two valves at both sides of the chamber under pressure changes in microchannels generated by repeated vertical movements of linear actuators. The volume of this system including the chip and the supporting device is 0.65 cubic decimeters, which is much smaller than that of reported systems with a volume of at least 12 cubic decimeters, and the weight of this system is only 0.56 kg, making it possible for point-of-care testing. We carry out an immunoassay of folic acid on chip, and the results show satisfactory reproducibility with acceptable coefficients of variation. We determine 163 clinical human serum samples for folic acid. Furthermore, we detect transferrin, cobalamin and folic acid simultaneously on one chip with both sandwich and competitive binding immunoassay methods. We anticipate that this on-chip valve and pump can be applied in immunoassays and other biosensing applications.

Corrigendum: Adenosine kinase on deoxyribonucleic acid methylation: Adenosine receptor-independent pathway in cancer therapy.

[This corrects the article DOI: 10.3389/fphar.2022.908882.].

Moderate Hyperkalemia Regulates Autophagy to Reduce Cerebral Ischemia-Reperfusion Injury in a CA/CPR Rat Model.

BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) can cause irreversible brain damage and autophagy has been implicated in the pathophysiology. Increasing serum potassium (K+) levels reduces CIRI, but the relationship between its protective mechanism and autophagy is unclear. In this study, we aimed to find the optimal degree of raising serum (K+) and to investigate the relationship between high (K+) and autophagy and the underlying mechanisms in a cardiac arrest/cardiopulmonary resuscitation (CA/CPR) rat model. METHODS: Sprague Dawley (SD) rats were divided into four groups: S group, N group, P group, and Q group. The rats S group and N group were administered saline. The rats P group and Q group were administered 640 mg/kg of potassium chloride (KCl) continuously pumped at 4 mL/h (21.3 mg/(kg·min) and divided according to the electrocardiogram (ECG) changes during the administration of KCl. After 24-h of resuscitation, neural damage was assessed by measuring neurological deficit score (NDS), oxidative stress markers, and pathological staining of the cerebral cortex. The level of autophagy and the expression of mTOR-ULK1-Beclin1 pathway-related proteins were evaluated using transmission electron microscopy (TEM), immunostaining, and western blotting. RESULTS: Our results revealed that high (K+) improved NDS and decreased the oxidative stress markers. The autophagosomes, autolysosomes, and lysosomes were decreased following treatment KCl. Furthermore, the levels of micro-tubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ, Unc-51-like kinase 1 (ULK1), and Beclin1 were decreased, whereas mTOR expression was increased in the cortex. CONCLUSION: The results demonstrated that moderate hyperkalemia could alleviate autophagy after CIRI via regulating the mTOR-ULK1-Beclin1 pathway.

Design, synthesis and evaluation of the anti-breast cancer activity of 1,3-oxazolo[4,5-d]pyrimidine and 1,3-oxazolo[5,4-d]pyrimidine derivatives.

A series of 1,3-oxazolo[4,5-d]pyrimidine and 1,3-oxazolo[5,4-d]pyrimidine derivatives were synthesized and functionalized in this study. The obtained compounds were tested against breast cancer cell lines of the NCI subpanel, followed by further analysis using the COMPARE algorithm from the Therapeutics Development Program, NCI. All synthesized derivatives displayed activity against most cell lines in the range of micromolar concentrations in terms of all parameters studied. Oxazolopyrimidine 5 exhibited the highest antitumor activity. A standard COMPARE analysis of the compounds showed that the vectors of the cytotoxic activity of derivatives 10 and 11 displayed a close to very high correlation with tamoxifen, and oxazolopyrimidine 13 displayed a very high correlation with the same drug. Five derivatives (2, 4, 6, 11 and 13) showed a high correlation with aclacinomycin A in the TGI vector. At the same time, compound 1 effectively suppressed ADK in cultured MDA-MB 231 cell lines, indicating that ADK is one of its targets through which it exerts anticancer properties. Based on molecular docking results, the possible binding mode of oxazolopyrimidine 1 to ADK has been suggested.

Arousal effect of caffeine depends on adenosine A2A receptors in the shell of the nucleus accumbens.

Caffeine, the most widely used psychoactive compound, is an adenosine receptor antagonist. It promotes wakefulness by blocking adenosine A(2A) receptors (A(2A)Rs) in the brain, but the specific neurons on which caffeine acts to produce arousal have not been identified. Using selective gene deletion strategies based on the Cre/loxP technology in mice and focal RNA interference to silence the expression of A(2A)Rs in rats by local infection with adeno-associated virus carrying short-hairpin RNA, we report that the A(2A)Rs in the shell region of the nucleus accumbens (NAc) are responsible for the effect of caffeine on wakefulness. Caffeine-induced arousal was not affected in rats when A(2A)Rs were focally removed from the NAc core or other A(2A)R-positive areas of the basal ganglia. Our observations suggest that caffeine promotes arousal by activating pathways that traditionally have been associated with motivational and motor responses in the brain.

[Expression, purification and application of EsxB protein in Staphylococcus aureus].

OBJECTIVE: This study aimed to establish the method of expression and purification of EsxB protein, explore the EsxB antibody-positive Staphylococcus aureus (S. aureus) clinical infection status and relevance of drug resistance. METHODS: Constructed EsxB prokaryotic expression system by homologous recombination, Ni(2+) column was used to purify EsxB protein; and then ELISA was used to detect the anti-EsxB antibodies in serum of 78 patients with S. aureus infection; antimicrobial susceptibility of related S. aureus strains by automatic bacterial identification analyzer. RESULTS: EsxB prokaryotic protein expression system was constructed and EsxB protein was purified successfully; anti-EsxB antibodies were present in the serum of patients with S. aureus infection up to 28.21% (22/78). The proportion of multi-drug resistant and Methicillin-resistant S. aureus strains isolated from anti-EsxB antibodies positive patients were 100.0% (22/22), 77.3% (17/22), respectively, which were statistically higher than those strains isolated from anti-EsxB antibody-negative patients (35.7% (20/56) and 21.4% (12/56), respectively) (all P values < 0.01). CONCLUSION: Method for expression and purification of EsxB protein was established. All the S. aureus strains isolated from EsxB antibody-positive patients were multidrug resistant strains and most of them were resistant to methicillin.

[Dietary supplement use and the related factors among infants and young children in Chengdu].

OBJECTIVE: To investigate the prevalence of dietary supplement use and relevant factors among infants and young children in the urban area of Chengdu. METHODS: 257 infants and young children aged 0-3 years and their parents were selected by simple random sampling. Questionnaires were used to collect information about family characteristic and the use of dietary supplements of infants and young children in the past month. Logistic regression analysis was used to explore the related factors. RESULTS: The prevalence of dietary supplements use among infants and young children was 87.5%. The main types of supplements included vitamin D, vitamin A, calcium, zinc and iron, with the using prevalence of 76.3%, 75.5%, 33.5%, 16.3%, 3.9%, respectively. The prevalence of other types supplement use were different by age of children (P < 0.05), in addition to iron. The prevalence of calcium and zinc supplements use were gradually increased with age increasing. Vitamin D and vitamin A supplements use in 6 - 11 months age group were higher than other children. Supplement use was associated with the attitude of the parents on infant and early childhood nutrition (OR = 0.186, P < 0.05) and the growth and development status of children( OR = 2.317, P < 0.05). CONCLUSION: Dietary supplement was used widely among infants and young children in Chengdu. It should pay more attention to improve professional guidance on dietary supplement.

Adenosine Kinase on Deoxyribonucleic Acid Methylation: Adenosine Receptor-Independent Pathway in Cancer Therapy.

Methylation is an important mechanism contributing to cancer pathology. Methylation of tumor suppressor genes and oncogenes has been closely associated with tumor occurrence and development. New insights regarding the potential role of the adenosine receptor-independent pathway in the epigenetic modulation of DNA methylation offer the possibility of new interventional strategies for cancer therapy. Targeting DNA methylation of cancer-related genes is a promising therapeutic strategy; drugs like 5-Aza-2'-deoxycytidine (5-AZA-CdR, decitabine) effectively reverse DNA methylation and cancer cell growth. However, current anti-methylation (or methylation modifiers) are associated with severe side effects; thus, there is an urgent need for safer and more specific inhibitors of DNA methylation (or DNA methylation modifiers). The adenosine signaling pathway is reported to be involved in cancer pathology and participates in the development of tumors by altering DNA methylation. Most recently, an adenosine metabolic clearance enzyme, adenosine kinase (ADK), has been shown to influence methylation on tumor suppressor genes and tumor development and progression. This review article focuses on recent updates on ADK and its two isoforms, and its actions in adenosine receptor-independent pathways, including methylation modification and epigenetic changes in cancer pathology.

A survey of COVID-19 detection and prediction approaches using mobile devices, AI, and telemedicine.

Since 2019, the COVID-19 pandemic has had an extremely high impact on all facets of the society and will potentially have an everlasting impact for years to come. In response to this, over the past years, there have been a significant number of research efforts on exploring approaches to combat COVID-19. In this paper, we present a survey of the current research efforts on using mobile Internet of Thing (IoT) devices, Artificial Intelligence (AI), and telemedicine for COVID-19 detection and prediction. We first present the background and then present current research in this field. Specifically, we present the research on COVID-19 monitoring and detection, contact tracing, machine learning based approaches, telemedicine, and security. We finally discuss the challenges and the future work that lay ahead in this field before concluding this paper.

Adenosine-A2A Receptor Signaling Plays a Crucial Role in Sudden Unexpected Death in Epilepsy.

Adenosinergic activities are suggested to participate in SUDEP pathophysiology; this study aimed to evaluate the adenosine hypothesis of SUDEP and specifically the role of adenosine A2A receptor (A2AR) in the development of a SUDEP mouse model with relevant clinical features. Using a combined paradigm of intrahippocampal and intraperitoneal administration of kainic acid (KA), we developed a boosted-KA model of SUDEP in genetically modified adenosine kinase (ADK) knockdown (Adk+/-) mice, which has reduced ADK in the brain. Seizure activity was monitored using video-EEG methods, and in vivo recording of local field potential (LFP) was used to evaluate neuronal activity within the nucleus tractus solitarius (NTS). Our boosted-KA model of SUDEP was characterized by a delayed, postictal sudden death in epileptic mice. We demonstrated a higher incidence of SUDEP in Adk+/- mice (34.8%) vs. WTs (8.0%), and the ADK inhibitor, 5-Iodotubercidin, further increased SUDEP in Adk+/- mice (46.7%). We revealed that the NTS level of ADK was significantly increased in epileptic WTs, but not in epileptic Adk+/- mutants, while the A2AR level in NTS was increased in epileptic (WT and Adk+/-) mice vs. non-epileptic controls. The A2AR antagonist, SCH58261, significantly reduced SUDEP events in Adk+/- mice. LFP data showed that SCH58261 partially restored KA injection-induced suppression of gamma oscillation in the NTS of epileptic WT mice, whereas SCH58261 increased theta and beta oscillations in Adk+/- mutants after KA injection, albeit with no change in gamma oscillations. These LFP findings suggest that SCH58261 and KA induced changes in local neuronal activities in the NTS of epileptic mice. We revealed a crucial role for NTS A2AR in SUDEP pathophysiology suggesting A2AR as a potential therapeutic target for SUDEP risk prevention.

Comprehensive analysis of the correlation between GSTM1 and tumor immunity in colon cancer.

Background: Glutathione S-transferase mu 1 (GSTM1) is one of the major glutathione conjugation enzymes. Its expression and activity have been suggested to correlate with the occurrence of colon cancer; however, the role of GSTM1 in tumor immunity remains unclear. Methods: Relevant data downloaded from The Cancer Genome Atlas (TCGA), Clinical Proteomic Tumor Analysis Consortium (CPTAC), and Human Protein Atlas (HPA) was used to perform a multi-dimensional expression analysis of GSTM1 in colon adenocarcinoma (COAD). The correlation between GSTM1 and tumor immunity was analyzed with multiple online tools. Then protein-protein interaction (PPI) network and functional enrichment analyses of GSTM1-associated immunomodulators were performed. Further, we developed the Cox regression model based on the GSTM1-related immunomodulators. Finally, a GSTM1-based clinical nomogram and a calibration curve was established to predict the probability and accuracy of long-term survival. Result: GSTM1 was significantly downregulated in COAD versus normal tissues. Infiltration levels of B cells, CD8+ T cells, and dendritic cells were closely correlated to GSTM1 gene copy number deletion, and GSTM1 expression levels in COAD positively correlated with dendritic cell, B cell, neutrophil, and macrophage infiltration. Functional enrichment analysis indicated 36 GSTM1-related immunomodulators are involved in immune-related pathways of regulating T cell activation and lymphocytic activation. A 2-gene prognostic risk signature based on the 36 GSTM1-related immunomodulators was built using the Cox regression model, and the risk signature in combination with stage had an area under the curve (AUC) value of 0.747 by the receiver operating characteristic method. patients with higher risk scores-calculated based on 2 gene prognostic risk characteristics and further identified as an independent prognostic factor-were associated with worse survival using the Kaplan-Meier analysis. Together, the clinical nomogram and calibration curve based on GSTM1 suggested a good prediction accuracy for long-term survival probability. Conclusions: Our study provided evidence supporting the significant role of GSTM1 in COAD immunity and suggests GSTM1 as a potential novel target for COAD immunotherapy.

Manipulation of adenosine kinase affects sleep regulation in mice.

Sleep and sleep intensity are enhanced by adenosine and its receptor agonists, whereas adenosine receptor antagonists induce wakefulness. Adenosine kinase (ADK) is the primary enzyme metabolizing adenosine in adult brain. To investigate whether adenosine metabolism or clearance affects sleep, we recorded sleep in mice with engineered mutations in Adk. Adk-tg mice overexpress a transgene encoding the cytoplasmic isoform of ADK in the brain but lack the nuclear isoform of the enzyme. Wild-type mice and Adk(+/-) mice that have a 50% reduction of the cytoplasmic and the nuclear isoforms of ADK served as controls. Adk-tg mice showed a remarkable reduction of EEG power in low frequencies in all vigilance states and in theta activity (6.25-11 Hz) in rapid eye movement (REM) sleep and waking. Adk-tg mice were awake 58 min more per day than wild-type mice and spent significantly less time in REM sleep (102 ± 3 vs 128 ± 3 min in wild type). After sleep deprivation, slow-wave activity (0.75-4 Hz), the intensity component of non-rapid eye movement sleep, increased significantly less in Adk-tg mice and their slow-wave energy was reduced. In contrast, the vigilance states and EEG spectra of Adk(+/-) and wild-type mice did not differ. Our data suggest that overexpression of the cytoplasmic isoform of ADK is sufficient to alter sleep physiology. ADK might orchestrate neurotransmitter pathways involved in the generation of EEG oscillations and regulation of sleep.