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The Slack channel (KCNT1, Slo2.2) is a sodium-activated and chloride-activated potassium channel that regulates heart rate and maintains the normal excitability of the nervous system. Despite intense interest in the sodium gating mechanism, a comprehensive investigation to identify the sodium-sensitive and chloride-sensitive sites has been missing. In the present study, we identified two potential sodium-binding sites in the C-terminal domain of the rat Slack channel by conducting electrophysical recordings and systematic mutagenesis of cytosolic acidic residues in the rat Slack channel C terminus. In particular, by taking advantage of the M335A mutant, which results in the opening of the Slack channel in the absence of cytosolic sodium, we found that among the 92 screened negatively charged amino acids, E373 mutants could completely remove sodium sensitivity of the Slack channel. In contrast, several other mutants showed dramatic decreases in sodium sensitivity but did not abolish it altogether. Furthermore, molecular dynamics (MD) simulations performed at the hundreds of nanoseconds timescale revealed one or two sodium ions at the E373 position or an acidic pocket composed of several negatively charged residues. Moreover, the MD simulations predicted possible chloride interaction sites. By screening predicted positively charged residues, we identified R379 as a chloride interaction site. Thus, we conclude that the E373 site and the D863/E865 pocket are two potential sodium-sensitive sites, while R379 is a chloride interaction site in the Slack channel.SIGNIFICANCE STATEMENTThe research presented here identified two distinct sodium and one chloride interaction sites located in the intracellular C-terminal domain of the Slack (Slo2.2, KCNT1) channel. Identification of the sites responsible for the sodium and chloride activation of the Slack channel sets its gating property apart from other potassium channels in the BK channel family. This finding sets the stage for future functional and pharmacological studies of this channel.

Small-conductance Ca2+-activated potassium channels subtype 2 (KCa2.2, also called SK2) are operated exclusively by a Ca2+-calmodulin gating mechanism. Heterozygous genetic mutations of KCa2.2 channels have been associated with autosomal dominant neurodevelopmental disorders including cerebellar ataxia and tremor in humans and rodents. Taking advantage of these pathogenic mutations, we performed structure-function studies of the rat KCa2.2 channel. No measurable current was detected from HEK293 cells heterologously expressing these pathogenic KCa2.2 mutants. When coexpressed with the KCa2.2_WT channel, mutations of the pore-lining amino acid residues (I360M, Y362C, G363S, and I389V) and two proline substitutions (L174P and L433P) dominant negatively suppressed and completely abolished the activity of the coexpressed KCa2.2_WT channel. Coexpression of the KCa2.2_I289N and the KCa2.2_WT channels reduced the apparent Ca2+ sensitivity compared with the KCa2.2_WT channel, which was rescued by a KCa2.2 positive modulator.

PLA MBs with high filtration efficiency and antibacterial activity were prepared by reducing viscosity by irradiation and blending ZnO and SiO2.

Castanopsis honey is a high-quality, phenolic acid and flavonoid-rich honey with good antioxidant capacity, and has good anti-inflammatory effects by regulation of NF-κB pathway.

The CRISPR-Cas system is a highly adaptive and RNA-guided immune system found in bacteria and archaea, which has applications as a genome editing tool and is a valuable system for studying the co-evolutionary dynamics of bacteriophage interactions. Here introduces CRISPRimmunity, a new web server designed for Acr prediction, identification of novel class 2 CRISPR-Cas loci, and dissection of key CRISPR-associated molecular events. CRISPRimmunity is built on a suite of CRISPR-oriented databases providing a comprehensive co-evolutionary perspective of the CRISPR-Cas and anti-CRISPR systems. The platform achieved a high prediction accuracy of 0.997 for Acr prediction when tested on a dataset of 99 experimentally validated Acrs and 676 non-Acrs, outperforming other existing prediction tools. Some of the newly identified class 2 CRISPR-Cas loci using CRISPRimmunity have been experimentally validated for cleavage activity in vitro. CRISPRimmunity offers the catalogues of pre-identified CRISPR systems to browse and query, the collected resources or databases to download, a well-designed graphical interface, a detailed tutorial, multi-faceted information, and exportable results in machine-readable formats, making it easy to use and facilitating future experimental design and further data mining. The platform is available at http://www.microbiome-bigdata.com/CRISPRimmunity. Moreover, the source code for batch analysis are published on Github (https://github.com/HIT-ImmunologyLab/CRISPRimmunity).

Potassium (K) is an essential electrolyte for cellular functions in living organisms, and disturbances in K+ homeostasis could lead to various chronic diseases (e.g. hypertension, cardiac disease, diabetes, and bone health). However, little is known about the natural distribution of stable K isotopes in mammals and its application to investigate the bodily homeostasis and/or as a biomarker for diseases. Here, we measured K isotopic compositions (δ41K, per mil deviation of 41K/39K from the NIST SRM 3141a standard) of brain, liver, kidney, and red blood cells (RBCs) from 10 mice (five females and five males) with three different genetic backgrounds. Our results reveal that different organs and RBCs have distinct K isotopic signatures. Specifically, the RBCs have heavy K isotopes enrichment with δ41K ranging from 0.67 to 0.08 ‰, while the brains show lighter K isotopic compositions with δ41K ranging from -1.13 to -0.09 ‰ compared to the livers (δ41K = -0.12 ± 0.58 ‰) and kidneys (δ41K = -0.24 ± 0.57 ‰). We found that the K isotopic and concentration variability is mostly controlled by the organs, with a minor effect of the genetic background and sex. Our study suggest that the K isotopic composition could be used as a biomarker for changes in K+ homeostasis and related diseases such as hypertension, cardiovascular, and neurodegenerative diseases.

Background: Biliary atresia (BA) is a destructive, obliterative cholangiopathy characterized by progressive fibro-inflammatory disorder and obliteration of intra- and extrahepatic bile ducts. The Jagged1 (JAG1) gene mutations have been found in some isolated BA cases. We aim to explore the association of common variants in JAG1 with isolated BA risk in the Chinese Han population.Methods: We genotyped 31 tag single nucleotide polymorphisms covering the JAG1 gene region in 333 BA patients and 1,665 healthy controls from the Chinese population, and performed case-control association analysis. The expression patterns of JAG1 homologs were investigated in zebrafish embryos, and the roles of jag1a and jag1b in biliary development were examined by morpholino knockdown in zebrafish.Results: Single nucleotide polymorphisms rs6077861 [PAllelic = 1.74 × 10−4, odds ratio = 1.78, 95% confidence interval: 1.31–2.40] and rs3748478 (PAllelic = 5.77 × 10−4, odds ratio = 1.39, 95% confidence interval: 1.15–1.67) located in the intron region of JAG1 showed significant associations with BA susceptibility. The JAG1 homologs, jag1a and jag1b genes were expressed in the developing hepatobiliary duct of zebrafish, especially at 72 and 96 h postfertilization. Knockdown of both jag1a and jag1b led to poor biliary secretion, sparse intrahepatic bile duct network and smaller or no gallbladders compared with control embryos in the zebrafish model.Conclusion: Common genetic variants of JAG1 were associated with BA susceptibility. Knockdown of JAG1 homologs led to defective intrahepatic and extrahepatic bile ducts in zebrafish. These results suggest that JAG1 might be implicated in the etiology of BA.

Overexpression of the thymidine phosphorylase (TP) enzyme induces angiogenesis, which eventually leads to metastasis and tumor growth. The crucial role of TP in cancer development makes it an important target for anticancer drug discovery. Currently, there is only one US-FDA-approved drug, i.e., Lonsurf, a combination of trifluridine and tipiracil, for the treatment of metastatic colorectal cancer. Unfortunately, numerous adverse effects are associated with its use, such as myelosuppression, anemia, and neutropenia. Since the last few decades, the discovery of new, safe, and effective TP inhibitory agents has been rigorously pursued. In the present study, we evaluated a series of previously synthesized dihydropyrimidone derivatives 1–40 for their TP inhibitory potential. Compounds 1, 12, and 33 showed a good activity with IC50 = 314.0 ± 0.90, 303.5 ± 0.40, and 322.6 ± 1.60 µM, respectively. The results of mechanistic studies revealed that compounds 1, 12, and 33 were the non-competitive inhibitors. These compounds were also evaluated for cytotoxicity against 3T3 (mouse fibroblast) cells and were found to be non-cytotoxic. Finally, the molecular docking suggested the plausible mechanism of non-competitive inhibition of TP. The current study thus identifies some dihydropyrimidone derivatives as potential inhibitors of TP, which can be further optimized as leads for cancer treatment.

Na Li, Meng Cui, Yumei Wang Department of Hospice Care, Shengjing Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of ChinaCorrespondence: Meng Cui; Yumei Wang, Department of Hospice Care, Shengjing Hospital of China Medical University, No. 39 Huaxiang Road, Tiexi District, Shenyang, Liaoning, People’s Republic of China, Tel +86 18940254587 ; +86 18940251490, Email meng_cui@hotmail.com; m18940251490@163.comBackground: Dyspnoea affects a considerable percentage of patients with terminal-stage cancer, and clinical guidelines recommend palliative sedation for patients with refractory dyspnoea. Midazolam is currently the most commonly used sedative; however, it can cause serious adverse reactions, such as respiratory/circulatory depression. Hence, there is a need for an alternative sedative. Dexmedetomidine (DEX) is a promising alternative as its “awake sedation” effect; however, little is known regarding its use in patients with end-stage dyspnoea. Therefore, the aim of this study was to determine the safety and usefulness of DEX for palliative sedation of patients with refractory dyspnoea.Methods: This retrospective study included patients with terminal-stage cancer who received DEX for palliative sedation owing to refractory dyspnoea in the hospice ward from January 2018 to October 2022. We analysed their general data, dyspnoea conditions, sedation details, sedative treatment effect, dyspnoea relief, and changes in vital signs before and after sedation, via paired t-tests.Results: We included 17 patients with terminal-stage cancer who received DEX palliative sedation at a dose of 0.2– 0.9 μg/kg·h for refractory dyspnoea, among whom 6 (35%) received a loading dose of 1 μg/kg in 10 min. After 1 h of sedation and at the maximum sedation dose, the Respiratory Distress Observation Scale and Richmond Agitation-Sedation Scale scores decreased significantly compared with those before sedation (all P< 0.001), as did the respiratory rate (P=0.024 and P=0.008, respectively). The heart rate and blood oxygen saturation did not significantly change, whereas the systolic and diastolic blood pressure after 1 h of sedation were significantly lower than those before sedation (both P=0.015).Conclusion: DEX is a promising palliative sedative for patients with terminal-stage cancer, as it safely relieved the symptoms of refractory dyspnoea without inducing serious adverse reactions. Therefore, DEX may greatly enhance the quality of life for patients with terminal-stage cancer.Keywords: palliative sedation, dexmedetomidine, dyspnoea, palliative care, terminal-stage cancer

The early-stage subduction records of the Paleo-Asian ocean are poorly preserved in the eastern segment of the Central Asian Orogenic Belt (CAOB), which hinders constraints on the evolution of the whole CAOB. This study presents new age data and zircon Hf-O isotopes as well as bulk-rock geochemistry of the Wuxing mafic-ultramafic complex in the Xingkai massif in northeastern China, which has been identified as Alaskan-type complex with aspects of field occurrence, petrological and mineral assemblages, and mineral chemistry in our recently published work. The results indicate that the complex formed mainly between 517 Ma and 510 Ma with a lithological sequence of Sanying clinopyroxenite (517 Ma), then Sanying gabbro (514 Ma), and Erying hornblendite (513 Ma), and finally Erying hornblende clinopyroxenite (510 Ma). The lithological formation sequence is consistent with the intrusive relations between lithological phases and their irrelevant major element compositions and variable trace element patterns of the bulk rocks. The εHf(t) values of zircon in two samples in this study vary from from +4.45 to +7.61 and from −11.8 to +4.42, respectively and tend to be more depleted with age. These features suggest that the Wuxing complex was a product of long-term arc magmatism and experienced significant ancient crustal assimilation in early-stage magmas and negligible contamination in later ones. The presence of 1222 Ma and 706 Ma inherited zircon grains implies existence of Proterozoic basement in the Xingkai massif and its continental arc setting in Cambrian. The Wuxing complex is the oldest Alaskan-type complex found so far in the entire CAOB and is a good witness of the Paleo-Asian oceanic subduction in the easternmost CAOB. The older age of the Wuxing complex compared to the regional Hongqiling intrusion is also compatible with its Alaskan-type nature and platinum-group element mineralization, which are distinct to the Permian-Triassic Ni-Cu sulfide deposit-hosting maficultramafic intrusions in the southern CAOB.

Green wearable electronics are attracting increasing attention to eliminate harmful byproducts generated by traditional devices. Although various degradable materials have been explored for green wearable electronics, the development of degradable elastomers with integrated characteristics of low modulus, self-adhesion, high resilient, and low hysteresis remains challenging. In this work, a degradable elastomer poly(1,8-octanediol

BACKGROUND AND PURPOSE The 5-HT receptor (5-HTR) subtypes 5-HT2A and 5-HT2C are important neurotherapeutic targets, though, obtaining selectivity over 5-HT2B and closely related histamine H1 Rs is challenging. Here, we delineated molecular determinants of selective binding to 5-HT2A and 5-HT2C Rs for novel 4-phenyl-2-dimethylaminotetralins (4-PATs). EXPERIMENTAL APPROACH We synthesized 42 novel 4-PATs with halogen or aryl moieties at the C(4)-phenyl meta position. Affinity, function, molecular modeling, and 5-HT2A R mutagenesis studies were undertaken to understand structure-activity relationships at 5-HT2 -type and H1 Rs. Lead 4-PAT-type selective 5-HT2A /5-HT2C R inverse agonists were compared to pimavanserin, a selective 5-HT2A /5-HT2C R inverse agonist approved to treat psychoses, in the mouse head twitch response, and locomotor activity assays, as models relevant to antipsychotic drug development. KEY RESULTS Most 4-PAT diastereomers in the (2S,4R)-configuration bound non-selectively to 5-HT2A , 5-HT2C, and H1 Rs, with >100-fold selectivity over 5-HT2B Rs, whereas, diastereomers in the (2R,4R)-configuration bound preferentially to 5-HT2A over 5-HT2C Rs and had >100-fold selectivity over 5-HT2B and H1 Rs. Results suggest that G2385.42 and V2355.39 in 5-HT2A Rs (conserved in 5-HT2C Rs) are important for high affinity binding, whereas, interactions with T1945.42 and W1584.56 determine H1 R affinity. The 4-PAT (2S,4R)-2k, a potent and selective 5-HT2A /5-HT2C R inverse agonist, had activity like pimavanserin in the mouse head-twitch response assay, but was distinct in not suppressing locomotor activity. CONCLUSIONS AND IMPLICATIONS We provide evidence that the novel 4-PAT chemotype can yield selective 5-HT2A /5-HT2C R inverse agonists for antipsychotic drug development by optimizing ligand-receptor interactions in transmembrane domain 5. We also show that chirality can be exploited to attain selectivity over H1 Rs which may circumvent sedative effects.

Textile pressure sensors capable of withstanding high temperatures have attracted increasing interest due to their potential applications in harsh conditions. In this work, a textile pressure sensor with high-temperature resistance was realized based on multi-wall carbon nanotubes (MWCNTs) and quartz fabrics. The textile pressure sensors exhibited low fatigue over 20 000 cyclic loadings. Owing to the high-temperature resistance of MWCNTs and quartz fabrics, the textile sensor can work at temperatures up to 300 °C and can maintain good sensitivity after calcination at 900 °C in N

The hardness and high abrasiveness of shale formations pose great challenges for improving drilling performance with deeper and more complex unconventional gas-reservoir formations explored and developed in the Sichuan Basin of China. The work discussed a workflow of data-driven drilling-parameter optimization based on machine learning. The algorithm was used to construct the correlation among drilling parameters, lithology change, vibration, bit wear, and hole cleaning, dynamically optimize rock-breaking efficiency, and integrate with the rig control system.The data-driven drilling optimization workflow consisted of exploration mode, learning mode, and application mode. The exploration phase trained a linear model at a certain frequency during data collection and updates the increasing trend of rate of penetration (ROP) in real time after starting the workflow. Based on the trend, the direction for further exploration was given. The system entered the learning mode after sufficient exploration to learn the exact functional relationship between ROP/MSE (mechanical specific energy) and operation parameters in the current explored data queue. Current optimal operation parameters were presented based on the function relationship. Then the workflow entered the application mode, maintained the current optimal operating parameters, and kept the efficient rock-breaking state. The workflow constantly monitored the micro-interval ROP and bit energy output in the application mode. When drilling performance was under expectation, the workflow automatically evaluated new conditions (e.g., formation change, Bottom hole assembly (BHA) vibration, and cuttings bed) and switched to the learning mode or exploration mode to adapt to changes in the current drilling state.The algorithm has been integrated with the rig control system, and the field test was carried out in well Ning 209H71-3, a shale-gas horizontal well in the Sichuan Basin. The test showed that the random forest and support vector machine algorithm could fit the nonlinear function relationship among drilling parameters, hole cleaning, and bit working performance, with properly optimized parameters presented. Besides, the workflow could evaluate the trends of ROP, MSE, depth of cutting (DOC), and stick-slips (SS) to capture the limiters for drilling performance, such as bit wear and lithology changes. Two modes integrated with global and local recommendations, and the optimal parameters have been provided to drillers in time. The field performance showed about 20% of ROP improvement with the recommended parameters along the horizontal section, and a 3,100-m horizontal section has been achieved.Machine learning algorithms were applied to drilling parameter recommendations with lower manual intervention. The novel workflow is not limited to bit type, downhole tools, rig equipment, etc. It has shown an outstanding drilling improvement in complex unconventional gas wells, which led the conventional drilling process to the automated era.

Saccharin is a commonly used artificial sweetener that exhibits both sweetening and sweet inhibition activities. The species‐dependent response towards saccharin and the interaction between saccharin and the sweet taste receptor T1R2/T1R3 remain elusive. In this study we used mismatched chimeras of T1R2 and T1R3 and calcium mobilization functional analysis to reveal a detailed species‐dependent response towards saccharin of human, squirrel monkey, and mouse sweet taste receptors. Our findings, combined with previous results by others, suggest multiple and complex interaction modes between saccharin and the sweet taste receptor, which are helpful guidelines for effective modulation of the sweet taste by sweetener/sweet inhibitors., Saccharin exhibits both sweetening and sweet inhibition activities. Besides the orthosteric binding site in T1R2 for activating the human sweet taste receptor T1R2/T1R3, it appears that there are at least two allosteric inhibitory sites located in T1R2 and T1R3 respectively to inhibit the receptor activity, suggesting multiple and complex interaction modes between saccharin and the heterodimeric sweet taste receptor T1R2/T1R3.

Background: Dyspnoea affects a considerable percentage of patients with terminal-stage cancer, and clinical guidelines recommend palliative sedation for patients with refractory dyspnoea, which cannot be alleviated with current therapies. Midazolam is currently the most commonly used sedative; however, it can cause serious adverse reactions, such as respiratory/circulatory depression. Hence, there is a need to discover an alternative sedative. Dexmedetomidine (DEX) has an “awake sedation” effect and is a promising alternative for palliative sedation; however, little is known regarding its use in patients with end-stage dyspnoea. Therefore, the aim of this study was to determine the safety and usefulness of DEX for palliative sedation of patients with refractory dyspnoea, especially focusing on those with terminal-stage cancer. Methods: This retrospective study included patients with terminal-stage cancer who received DEX for palliative sedation owing to refractory dyspnoea in the hospice ward from January 2018 to October 2022. We analysed their general data, dyspnoea conditions, sedation details, sedative treatment effect, dyspnoea relief, and changes in vital signs before and after sedation, via paired t-tests. Results: We included 17 patients with terminal-stage cancer who received DEX palliative sedation at a dose of 0.2-0.9 µg/kg·h for refractory dyspnoea, among whom 6 (35%) received a loading dose of 1 µg/kg in 10 min. After 1 h of sedation and at the maximum sedation dose, the Respiratory Distress Observation Scale and Richmond Agitation-Sedation Scale scores decreased significantly compared with those before sedation (all PP=0.024 and P=0.008, respectively). The heart rate and blood oxygen saturation did not significantly change after 1 h of sedation or at the maximum sedation dose, whereas the systolic and diastolic blood pressure after 1 h of sedation were significantly lower than those before sedation (both P=0.015); however, these findings were not observed at the maximum sedation dose. Conclusions: According to these results, DEX is a promising palliative sedative for patients with terminal-stage cancer, as it safely and permanently relieved the symptoms of refractory dyspnoea without inducing serious adverse reactions, such as respiratory depression. Therefore, DEX may greatly enhance the quality of life for patients with terminal-stage cancer.

We engineered and produced an ion channel blocking peptibody, that targets the acetylcholine-activated inwardly rectifying potassium current (I KACh ). Peptibodies are chimeric proteins generated by fusing a biologically active peptide with the fragment crystallizable (Fc) region of the human immunoglobulin G (IgG). The I KACh blocking peptibody was engineered as a fusion between the human IgG1 Fc fragment and the I KACh inhibitor tertiapinQ (TP), a 21-amino acid synthetic peptidotoxin, originally isolated from the European honey bee venom. The peptibody was purified from the culture supernatant of human embryonic kidney (HEK) cells transfected with the peptibody construct. We tested the hypothesis that the bioengineered peptibody is bioactive and a potent blocker of I KACh . In HEK cells transfected with Kir3.1 and Kir3.4, the molecular correlates of I KACh , patch clamp showed that the peptibody was ~300-fold more potent than TP. Molecular dynamics simulations suggested that the increased potency could be due to an increased stabilization of the complex formed by peptibody-Kir3.1/3.4 channels compared to tertiapin-Kir3.1/3.4 channels. In isolated mouse myocytes, the peptibody blocked carbachol (Cch)-activated I KACh in atrial cells but did not affect the potassium inwardly rectifying background current in ventricular myocytes. In anesthetized mice, the peptibody abrogated the bradycardic effects of intraperitoneal Cch injection. Moreover, in aged mice, the peptibody reduced the inducibility of atrial fibrillation, likely via blocking constitutively active I KACh . Bioengineered anti-ion channel peptibodies can be powerful and highly potent ion channel blockers, with the potential to guide the development of modulators of ion channels or antiarrhythmic modalities.

Ting Ye,1,&ast; Jia Feng,1,&ast; Meng Cui,2 Jia Yang,2 Xue Wan,1 Dan Xie,1 Jinbo Liu1 1Department of Laboratory Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, 646000, People’s Republic of China; 2Department of Laboratory Medicine, The Leshan People’s Hospital, Luzhou, Sichuan, 614000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ting Ye; Jinbo LiuDepartment of Laboratory Medicine, The Affiliated Hospital of Southwest Medical University, No. 25, Taiping Street, Jiangyang District, Luzhou, Sichuan, 646000, People’s Republic of ChinaTel +86-830-3165730Email yeting1103@163.com; liulab2019@163.comBackground: Myocardial infarction associated transcript (MIAT) is identified as a long chain non-coding RNA (lncRNA), which was associated with myocardial infarction susceptibility. While intense efforts have been made to elucidate the relationship between MIAT and carcinogenesis, the tumor immunoreaction of MIAT remains elusive. Thus, this study aimed to investigate the role of MIAT in the immunoregulation of breast cancer (BC) and further explore the better clinical significance.Methods: The differential expression of MIAT between BC and normal/adjacent tissues was compared using Wilcoxon rank sum test. The diagnostic and prognostic values of elevated MIAT expression in BC tissues were unveiled via receiver operating characteristic (ROC) analysis and KM-plotter analysis. Limma and edgeR package were used to identify differentially expressed genes (DEGs) and microRNAs (DEMs) from TCGA database respectively. A co-expression dataset was constructed to comprehensively understand the relationship between MIAT and DEGs based on the Pearson correlation coefficient. Furthermore, GO and KEGG analyses were conducted to predict the potential functions of MIAT. We next intersected immune-related genes (IRGs) from ImmPort database with MIAT-co-expressed genes to obtain MIAT-co-expressed IRGs, in order to construct MIAT-microRNA (miRNA)-mRNA network. And the correlation between MIAT and tumor-infiltrating immune cells (TICs) and immunophenoscore (IPS) analysis was analyzed by TIMER and CIBERSORT. Finally, the reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expression profiles of MIAT in serum samples.Results: The expression levels of MIAT were notably higher in BC than in normal or adjacent tissues. And MIAT expression could be used as a prognostic indicator of mortality risk in patients with BC in different aspects. Moreover, the enrichment analyses suggested that MIAT was strongly involved in BC immune response. In addition, TIMER database and CIBERSORT analyses indicated that MIAT was significantly correlated with 13 types of TICs (B cells, dendritic cells, neutrophils, CD8 T cells, CD4 memory resting T cells, CD4 memory activated T cells, gamma delta T cells, M1 macrophages, plasma cells, activated NK cells, monocytes, M2 macrophages, activated mast cells). Simultaneously, the IPS analysis implied that the higher the MIAT expression, the better the immunotherapy effect. The ROC curve analysis showed that the area under the curve (AUC) value of MIAT was 0.86 (sensitivity = 87.80%, specificity = 75.61%). And the high MIAT expression in serum was positive related to TNM stage (P = 0.032) and lymph node metastasis (P = 0.028).Conclusion: MIAT may be a valuable noninvasive diagnostic biomarker for BC and is associated with tumor-infiltrating immune cells in tumor microenvironment, suggesting MIAT as a potential target for future treatment of BC.Keywords: lncRNA MIAT, BC, biomarker, immune regulation, tumor-infiltrating, immune cells

BACKGROUND AND PURPOSE: In the activated state of small-conductance Ca(2+)-activated potassium (K(Ca) 2) channels, calmodulin interacts with the HA/HB helices and the S4-S5 linker. CyPPA potentiates K(Ca) 2.2a and K(Ca) 2.3 channel activity but not the K(Ca) 2.1 and K(Ca) 3.1 subtypes. EXPERIMENTAL APPROACH: Site-directed mutagenesis, patch-clamp recordings and in silico modeling were utilized to explore the structural determinants for the subtype-selective modulation of K(Ca) 2 channels by CyPPA. KEY RESULTS: Mutating residues in the HA (V420) and HB (K467) helices of K(Ca) 2.2a channels to their equivalent residues in K(Ca) 3.1 channels diminished the potency of CyPPA. CyPPA elicited prominent responses on mutant K(Ca) 3.1 channels with an arginine residue in the HB helix substituted for its equivalent lysine residue in the K(Ca) 2.2a channels (R355K). K(Ca) 2.1 channels harboring a three-amino-acid insertion upstream of the cognate R438 residues in the HB helix showed no response to CyPPA, whereas the deletion mutant (K(Ca) 2.1_ΔA434/Q435/K436) became sensitive to CyPPA. In molecular dynamics simulations, CyPPA docked between calmodulin C-lobe and the HA/HB helices widens the cytoplasmic gate of K(Ca) 2.2a channels. CONCLUSION AND IMPLICATIONS: Selectivity of CyPPA among K(Ca) 2 and K(Ca) 3.1 channel subtypes relies on the HA/HB helices.

Gallstones were examined for 58 patients in Jilin, Jilin. It was found that gallstones from only one who suffered from cholecystectomy were composed of 20 spheroidal stones and they rarely exhibited three different types of surface appearances. Six representative powder samples were analyzed for gallstones compositions and Mn2+ using X-ray diffraction (XRD)/Infrared (IR) and electron spin resonance (ESR), respectively. The results suggested that all gallstones from this patient were identified by XRD to be gallstones consisting mainly of cholesterol and CaCO3 (GCCC). They rarely exhibited three different kinds of surface appearances corresponding to different concentration of trace Mn2+ in calcite (CMn2+/CCal): 18 dark/light brown spheres with smooth surfaces and CMn2+/CCal = 0–6 μg/g/%, a yellowish-brown huge sphere with a rougher surface and CMn2+/CCal = 30 μg/g/%, and an ashy sphere composed of tens of microspheres with the roughest surface and CMn2+/CCal = 60 μg/g/%. The difference in surface appearance showed significant association with CMn2+/CCal, and its increase made the gallstone’s surface change from smooth to rough and to fade in color. The unbalanced and competitive Mn2+ accumulation could occur occasionally in individual stones owing to different affinities to Mn2+, resulting in the formation of a huge stone and an ashy sphere. These two aberrations caused by higher CMn2+/CCal played an important role in suppressing the crystalline growth of the majority of dark/light brown spheres. GCCC from a patient might have a prominent Mn2+ partitioning feature corresponding to different surface appearances.

Metallothionein (MT) is a multifunctional inducible protein in animals, plants, and microorganisms. MT is rich in cysteine residues (10-30%), can combine with metal ions, has a low molecular weight, and plays an essential biological role in various stages of the growth and development of organisms. Due to its strong ability to bind metal ions and scavenge free radicals, metallothionein has been used in medicine, health care, and other areas. Zinc is essential for plant growth, but excessive zinc (Zn) is bound to poison plants, and cadmium (Cd) is a significant environmental pollutant. A high concentration of cadmium can significantly affect the growth and development of plants and even lead to plant death. In this study, the human metallothionein gene

The Na-K-2Cl cotransporter-1 (NKCC1) is an electroneutral Na + -dependent transporter responsible for simultaneously translocating Na + , K + , and Cl − ions into cells. In human tissue, NKCC1 plays a critical role in regulating cytoplasmic volume, fluid intake, chloride homeostasis, and cell polarity. Here, we report four structures of human NKCC1 (hNKCC1), both in the absence and presence of loop diuretic (bumetanide or furosemide), using single-particle cryo–electron microscopy. These structures allow us to directly observe various novel conformations of the hNKCC1 dimer. They also reveal two drug-binding sites located at the transmembrane and cytosolic carboxyl-terminal domains, respectively. Together, our findings enable us to delineate an inhibition mechanism that involves a coupled movement between the cytosolic and transmembrane domains of hNKCC1.