Single Isotopologue for In-Sample Calibration and Absolute Quantitation by LC-MS/MS.

Targeted mass spectrometry (MS)-based absolute quantitative analysis has been increasingly used in biomarker discovery. The ability to accurately measure the masses by MS enabled the use of isotope-incorporated surrogates having virtually identical physiochemical properties with the target analytes as calibrators. Such a unique capacity allowed for accurate in-sample calibration. Current in-sample calibration uses multiple isotopologues or structural analogues for both the surrogate and the internal standard. Here, we simplified this common practice by using endogenous light peptides as the internal standards and used a mathematical deduction of "heavy matching light, HML" to directly quantify an endogenous analyte. This method provides all necessary assay performance parameters in the authentic matrix, including the lower limit of quantitation (LLOQ) and intercept of the calibration curve, by using only a single isotopologue of the analyte. This method can be applied to the quantitation of proteins, peptides, and small molecules. Using this method, we quantified the efficiency of heart tissue digestion and recovery using sodium deoxycholate as a detergent and two spiked exogenous proteins as mimics of heart proteins. The results demonstrated the robustness of the assay.

Construction and validation of a risk model of proteinuria in patients with omicron COVID-19: retrospective cohort study.

BACKGROUND: To explore the risk factors of proteinuria in Omicron variant patients and to construct and verify the risk predictive model. METHODS: 1091 Omicron patients who were hospitalized from August 2022 to November 2022 at Tianjin First Central Hospital were defined as the derivation cohort. 306 Omicron patients who were hospitalized from January 2022 to March 2022 at the same hospital were defined as the validation cohort. The risk factors of proteinuria in derivation cohort were screened by univariate and multivariate logistic regression analysis, and proteinuria predicting scoring system was constructed and the receiver operating characteristic(ROC)curve was drawn to test the prediction ability. The proteinuria risk model was externally validated in validation cohort. RESULTS: 7 factors including comorbidities, blood urea nitrogen (BUN), serum sodium (Na), uric acid (UA), C reactive protein (CRP) and vaccine dosages were included to construct a risk predictive model. The score ranged from -5 to 16. The area under the ROC curve(AUC) of the model was 0.8326(95% CI 0.7816 to 0.8835, p < 0.0001). Similarly to that observed in derivation cohort, the AUC is 0.833(95% CI 0.7808 to 0.9002, p < 0.0001), which verified good prediction ability and diagnostic accuracy in validation cohort. CONCLUSIONS: The risk model of proteinuria after Omicron infection had better assessing efficiency which could provide reference for clinical prediction of the risk of proteinuria in Omicron patients.

High-Entropy-Inspired Multicomponent Electrical Double Layer Structure Design for Stable Zinc Metal Anodes.

Regulating the electrical double layer (EDL) structure can enhance the cycling stability of Zn metal anodes, however, the effectiveness of this strategy is significantly limited by individual additives. Inspired by the high-entropy (HE) concept, we developed a multicomponent (MC) EDL structure composed of La3+, Cl-, and BBI anions by adding dibenzenesulfonimide (BBI) and LaCl3 additives into ZnSO4 electrolytes (BBI/LaCl3/ZnSO4). Specifically, La3+ ions accumulate within EDL to shield the net charges on the Zn surface, allowing more BBI anions and Cl- ions to enter this region. Consequently, this unique MC EDL enables Zn anodes to simultaneously achieve uniform electric field, robust SEI layer, and balanced reaction kinetics. Moreover, the synergistic parameter - a novel descriptor for quantifying collaborative improvement - was first proposed to demonstrates the synergistic effect between BBI and LaCl3 additives. Benefitting from these advantages, Zn metal anodes achieved a high reversibility of 99.5 % at a depth of discharge (DoD) of 51.3 %, and Zn|MnO2 pouch cells exhibited a stable cycle life of 100 cycles at a low N/P ratio of 2.9.

The pattern of cytokines expression and dynamic changes of renal function at 6 months in patients with Omicron COVID-19.

To analyze the dynamic changes of renal function longitudinally and investigate the cytokine profiles at 6 months in patients with Omicron COVID-19. Forty-seven patients with a proven diagnosis of Omicron COVID-19 from January to February 2022 attended a 6-month follow-up after discharge at Tianjin First Central Hospital. The demographic parameters, clinical features, and laboratory indexes were collected during hospitalization and 6 months after discharge. The serum cytokine levels at 6 months were also assessed. Patients were grouped according to with or without kidney involvement at admission. The levels of serum creatinine and estimated glomerular filtration rate (eGFR) were all normal both in the hospital and at follow-up. Whereas, compared with renal function in the hospital, serum creatinine levels at 6 months increased remarkably; meanwhile, eGFR decreased significantly in all patients. The serum levels of interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, and TNF-α and IFN-γ significantly decreased and TGF-ß remarkably increased in the kidney involvement group. The serum levels of IL-2 and IL-5 were positively correlated with age; contrarily, TGF-ß showed a negative correlation with aging. The younger was an independent risk factor of the higher TGF-ß levels. Omicron patients showed a decline in renal function at follow-up, reflecting the trend of CKD. Serum cytokine profiles were characterized with the majority of cytokines decreased and TGF-ß increased in the kidney involvement group; the latter may be used as a sign of CKD. The tendency of CKD is one of the manifestations of long COVID and deserves attention.

Long Shelf-Life Efficient Electrolytes Based on Trace l-Cysteine Additives toward Stable Zinc Metal Anodes.

The unstable anode/electrolyte interface (AEI) triggers the corrosion reaction and dendrite formation during cycling, hindering the practical application of zinc metal batteries. Herein, for the first time, l-cysteine (Cys) is employed to serve as an electrolyte additive for stabilizing the Zn/electrolyte interface. It is revealed that Cys additives tend to initially approach the Zn surface and then decompose into multiple effective components for suppressing parasitic reactions and Zn dendrites. As a consequence, Zn|Zn symmetric cells using trace Cys additives (0.83 mm) exhibit a steady cycle life of 1600 h, outperforming that of prior studies. Additionally, an average Coulombic efficiency of 99.6% for 250 cycles is also obtained under critical test conditions (10 mA cm-2 /5 mAh cm-2 ). Cys additives also enable Zn-V2 O5 and Zn-MnO2 full cells with an enhanced cycle stability at a low N/P ratio. More importantly, Cys/ZnSO4 electrolytes are demonstrated to be still effective after resting for half year, favoring the practical production.

Absolute Quantification of Nav1.5 Expression by Targeted Mass Spectrometry.

Nav1.5 is the pore forming α-subunit of the cardiac voltage-gated sodium channel that initiates cardiac action potential and regulates the human heartbeat. A normal level of Nav1.5 is crucial to cardiac function and health. Over- or under-expression of Nav1.5 can cause various cardiac diseases ranging from short PR intervals to Brugada syndromes. An assay that can directly quantify the protein amount in biological samples would be a priori to accurately diagnose and treat Nav1.5-associated cardiac diseases. Due to its large size (>200 KD), multipass transmembrane domains (24 transmembrane passes), and heavy modifications, Nav1.5 poses special quantitation challenges. To date, only the relative quantities of this protein have been measured in biological samples. Here, we describe the first targeted and mass spectrometry (MS)-based quantitative assay that can provide the copy numbers of Nav1.5 in cells with a well-defined lower limit of quantification (LLOQ) and precision. Applying the developed assay, we successfully quantified transiently expressed Nav1.5 in as few as 1.5 million Chinese hamster ovary (CHO) cells. The obtained quantity was 3 ± 2 fmol on the column and 3 ± 2 × 104 copies/cell. To our knowledge, this is the first absolute quantity of Nav1.5 measured in a biological sample.

Hispolon Methyl Ether, a Hispolon Analog, Suppresses the SRC/STAT3/Survivin Signaling Axis to Induce Cytotoxicity in Human Urinary Bladder Transitional Carcinoma Cell Lines.

Bladder cancer is a leading human malignancy worldwide. Signal transducer and activator of transcription (STAT) 3 is an oncogenic transcription factor commonly hyperactivated in most human cancers, including bladder cancer. Notably, preclinical evidence has validated STAT3 blockade as a promising therapeutic strategy for bladder cancer. Hispolon Methyl Ether (HME) is a structural analog of hispolon, an anticancer component of the medicinal mushroom Phellinus linteus. Thus far, HME's anticancer activity and mechanisms remain largely unknown. We herein report HME was cytotoxic, more potent than cisplatin, and proapoptotic to various human bladder transitional carcinoma cell lines. Of note, HME blocked STAT3 activation, evidenced by HME-elicited reduction in tyrosine 705-phosphorylated STAT3 levels constitutively expressed or induced by interleukin-6. Significantly, HME-induced cytotoxicity was abrogated in cells expressing a dominant-active STAT3 mutant (STAT3-C), confirming STAT3 blockage as a pivotal mechanism of HME's cytotoxic action. We further revealed that survivin was downregulated by HME, while its levels were rescued in STAT3-C-expressing cells. Moreover, survivin overexpression abolished HME-induced cytotoxicity, illustrating survivin as a central downstream mediator of STAT3 targeted by HME. Lastly, HME was shown to lower tyrosine 416-phosphorylated SRC levels, suggesting that HME inhibits STAT3 by repressing the activation of SRC, a STAT3 upstream kinase. In conclusion, we present the first evidence of HME's anti-bladder cancer effect, likely proceeding by evoking apoptosis through suppression of the antiapoptotic SRC/STAT3/survivin signaling axis.

MicroRNA-4461 derived from bone marrow mesenchymal stem cell exosomes inhibits tumorigenesis by downregulating COPB2 expression in colorectal cancer.

Colorectal cancer (CRC) is one of the main cause of cancer-related deaths. It's reported that bone marrow mesenchymal stem cells (BMSCs) affects tumor development through secreting exosomes. This study aims to investigate the function of BMSCs-derived exosome miR-4461 in CRC. The results of qRT-PCR showed that miR-4461 expression in DLD1, HCT116 and SW480 CRC cells and CRC tissues was lower than that in FHC cells and normal tissues, respectively. And COPB2 mRNA expression was negatively correlated with miR-4461. Western blot was used to detect COPB2 protein expression. Dual-luciferase reporter assay results revealed that miR-4461 targeted COPB2. Transwell assay and CCK-8 assay demonstrated that COPB2 knockdown inhibited HCT116 and SW480 cells proliferation, migration and invasion abilities. Furthermore, BMSCs-derived exosome miR-4461 downregulated COPB2 expression and inhibited HCT116 and SW480 cells migration and invasion. The findings demonstrated that miR-4461 could be a potential target for the diagnosis and treatment of colorectal cancer.

Biosensor-assisted transcriptional regulator engineering for Methylobacterium extorquens AM1 to improve mevalonate synthesis by increasing the acetyl-CoA supply.

Acetyl-CoA is not only an important intermediate metabolite for cells but also a significant precursor for production of industrially interesting metabolites. Methylobacterium extorquens AM1, a model strain of methylotrophic cell factories using methanol as carbon source, is of interest because it produces abundant coenzyme A compounds capable of directing to synthesis of different useful compounds from methanol. However, acetyl-CoA is not always efficiently accumulated in M. extorquens AM1, as it is located in the center of three cyclic central metabolic pathways. Here we successfully demonstrated a strategy for sensor-assisted transcriptional regulator engineering (SATRE) to control metabolic flux re-distribution to increase acetyl-CoA flux from methanol for mevalonate production in M. extorquens AM1 with introduction of mevalonate synthesis pathway. A mevalonate biosensor was constructed and we succeeded in isolating a mutated strain (Q49) with a 60% increase in mevalonate concentration (an acetyl-CoA-derived product) following sensor-based high-throughput screening of a QscR transcriptional regulator library. The mutated QscR-49 regulator (Q8*,T61S,N72Y,E160V) lost an N-terminal α-helix and underwent a change in the secondary structure of the RD-I domain at the C terminus, two regions that are related to its interaction with DNA. 13C labeling analysis revealed that acetyl-CoA flux was improved by 7% and transcriptional analysis revealed that QscR had global effects and that two key points, NADPH generation and fumC overexpression, might contribute to the carbon flux re-distribution. A fed-batch fermentation in a 5-L bioreactor for QscR-49 mutant yielded a mevalonate concentration of 2.67g/L, which was equivalent to an overall yield of 0.055mol acetyl-CoA/mol methanol, the highest yield among engineered strains of M. extorquens AM1. This work was the first attempt to regulate M. extorquens AM1 on transcriptional level and provided molecular insights into the mechanism of carbon flux regulation.

Characterization and comparison of proteomes of albino sea cucumber Apostichopus japonicus (Selenka) by iTRAQ analysis.

Sea cucumber is a commercially important marine organism in China. Of the different colored varieties sold in China, albino sea cucumber has the greatest appeal among consumers. Identification of factors contributing to albinism in sea cucumber is therefore likely to provide a scientific basis for improving the cultivability of these strains. In this study, two-dimensional liquid chromatography-tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification labeling was used for the first time to quantitatively define the proteome of sea cucumbers and reveal proteomic characteristics unique to albino sea cucumbers. A total of 549 proteins were identified and quantified in albino sea cucumber and the functional annotations of 485 proteins have been exhibited based on COG database. Compared with green sea cucumber, 12 proteins were identified as differentially expressed in the intestine and 16 proteins in the body wall of albino sea cucumber. Among them, 5 proteins were up-regulated in the intestine and 8 proteins were down-regulated in body wall. Gene ontology annotations of these differentially expressed proteins consisted mostly of 'biological process'. The large number of differentially expressed proteins identified here should be highly useful in further elucidating the mechanisms underlying albinism in sea cucumber.

Development of an Absolute Quantification Method for hERG Using PRM with Single Isotopologue in-Sample Calibration.

The human ether-à-go-go-related gene (KCNH2)-encoded protein hERG constitutes the α subunit of the Kv11.1 channel and contributes to the I kr current, which plays an important role in the cardiac action potential. Genetically and xenobiotically triggered malfunctions of hERG can cause arrhythmia. The expression of hERG in various study systems was assessed mainly as the fold change relative to the corresponding control. Here, we developed a simple and sensitive quantitation method using targeted mass spectrometry, i.e., the parallel reaction monitoring approach, to measure the absolute quantity of hERG in copy number. Such measurements do not require controls, and the obtained values can be compared with similar results for any other protein. To effectively avoid matrix effects, we used the heavy-match-light (HML) in-sample calibration approach that requires only a single isotopologue to achieve copy-number quantitation. No significant difference was observed in the results obtained by HML and by the classic standard addition in-sample calibration approach. Using four proteotypic peptides, we quantified the average number of copies of hERG in the HEK293T heterologous expression system as 3.6 ± 0.5 × 106 copies/cell, i.e., 1 million copies/cell for the fully assembled Kv11.1 channel.

Synergistic effect between S and Te enhancing the electrochemical behavior of heteroatomic TeS-x cathodes in aqueous Zn-TeS batteries.

Aqueous Zn-S batteries (AZSBs) have garnered increasing attention in the energy storage field owing to their high capacity, energy density, and cost effectiveness. Nevertheless, sulfur (S) cathodes face challenges, primarily stemming from sluggish reaction kinetics and the formation of an irreversible byproduct (SO42-) during the charge, hindering the progress of AZSBs. Herein, Te-S bonds within S-based cathodes were introduced to enhance electron and ion transport and facilitate the conversion reaction from zinc sulfide (ZnS) to S. This was achieved by constructing heteroatomic TeS-x@Ketjen black composite cathodes (HM-TeS-x@KB, where x  = 36, 9, and 4). The HM-TeS-9@KB electrode exhibits long-term cycling stability, maintaining a capacity decay rate of 0.1 % per cycle over 450 cycles at a current density of 10 A g-1. Crucially, through a combination of experimental data analysis and theoretical calculations, the impact mechanism of Te on the charge and discharge of S active materials within the HM-TeS-9@KB cathode in AZSBs was investigated. The presence of Te-S bonds boost the intrinsic conductivity and wettability of the HM-TeS-9@KB cathode. Furthermore, during the charge, the interaction of preferentially oxidized Te with S atoms within ZnS promotes the oxidation reaction from ZnS to S and suppresses the irreversible side reaction between ZnS and H2O. These findings indicate that the heteroatomization of chalcogen S molecules represents a promising approach for enhancing the electrochemical performance of S cathodes in AZSBs.

Afforestation-Induced Shifts in Soil Bacterial Diversity and Community Structure in the Saihanba Region.

Afforestation plays a pivotal role in ecosystem restoration, exemplified by the Saihanba Mechanized Forest Farm, the world's largest planted forest; however, the assembly mechanisms and interactions of soil microbial communities in such forests remain inadequately understood. This study aimed to elucidate the impact of different afforestation tree species, namely Larix gmelinii var. principis-rupprechtii, Picea asperata, and Pinus sylvestris var. mongolica, on soil bacterial diversity and community structure in comparison to grassland. Sixty soil samples were collected at a 20 cm depth, and high-throughput sequencing was employed to identify bacterial communities and assess their interactions with environmental factors. A total of 6528 operational taxonomic units (OTUs) were identified, with Solirubrobacter, Conexibacter, Bacillus, Massilia, Gaiella, Acidibacter, and Vicinamibacter being the dominant genera. Afforestation significantly impacted soil bacterial alpha diversity, with notable influence from key soil chemical properties, including available phosphorus (AP), cation exchange capacity (CEC), and the carbon-to-nitrogen ratio of soil organic matter (SOM-C/N). The Mantel test highlighted pH, the Normalized Difference Vegetation Index (NDVI), and spatial variable (dbMEM) as primary environmental factors influencing dominant bacterial genera. The bacterial community structure demonstrated deterministic homogeneous selection, wherein SOM-C/N emerged as a significant factor influencing the dissimilarity of soil bacterial communities. Furthermore, plantation soils exhibited a more complex network structure than grassland soil, highlighting the crucial role of bacterial communities in vegetation changes and providing valuable insights into their response to environmental factors during the reforestation process.

catena-Poly[[di-aqua-[µ2-4-(4-carb-oxy-phen-oxy)benzoato](µ2-4,4'-oxydibenzo-ato)praseodymium(III)] monohydrate].

In the title compound, {[Pr(C14H8O5)(C14H9O5)(H2O)2]·H2O} n , the Pr(III) cation is eight-coordinated by six carboxyl O atoms from both a monoanionic 4-(4-carb-oxy-phen-oxy)benzoate and a dianionic 4,4'-oxydibenzoate ligand (four bridging with two from a bidentate chelate inter-action), and two O-atom donors from water mol-ecules. A single water mol-ecule of solvation is also present. The complex units are linked through carboxyl O:O' bridges giving a two-dimensional sheet polymer lying parallel to (001). An overall three-dimensional network structure is generated through inter-molecular carb-oxy-lic acid and water O-H⋯O hydrogen bonds and weak C-H⋯O inter-actions.

2,3-Bis(furan-2-yl)pyrazino-[2,3-f][1,10]phenanthroline.

The mol-ecule of the title compound, C22H12N4O2, is located on a twofold rotation axis. The dihedral angle between the furan and pyrazine rings is 34.8 (7)°, and that between the furan rings is 46.92 (7)°. A π-π stacking interaction occurs between adjacent pyrazino[2,3-f][1,10]phenanthroline units, with an interplanar distance of 3.5862 (12) Å.

Hepatic Stellate Cell- and Liver Microbiome-Specific Delivery System for Dihydrotanshinone I to Ameliorate Liver Fibrosis.

Liver fibrosis is a major contributor to the morbidity and mortality associated with liver diseases, yet effective treatment options remain limited. Hepatic stellate cells (HSCs) are a promising target for hepatic fibrogenesis due to their pivotal role in disease progression. Our previous research has demonstrated the potential of Dihydrotanshinone I (DHI), a lipophilic component derived from the natural herb Salvia miltiorrhiza Bunge, in treating liver fibrosis by inhibiting the YAP/TEAD2 interaction in HSCs. However, the clinical application of DHI faces challenges due to its poor aqueous solubility and lack of specificity for HSCs. Additionally, recent studies have implicated the impact of liver microbiota, distinct from gut microbiota, on the pathogenesis of liver diseases. In this study, we have developed an HSC- and microbiome-specific delivery system for DHI by conjugating prebiotic-like cyclodextrin (CD) with vitamin A, utilizing PEG2000 as a linker (VAP2000@CD). Our results demonstrate that VAP2000@CD markedly enhances the cellular uptake in human HSC line LX-2 and enhances the deposition of DHI in the fibrotic liver in vivo. Subsequently, intervention with DHI-VAP2000@CD has shown a notable reduction in bile duct-like structure proliferation, collagen accumulation, and the expression of fibrogenesis-associated genes in rats subjected to bile duct ligation. These effects may be attributed to the regulation of the YAP/TEAD2 interaction. Importantly, the DHI-VAP2000@CD intervention has also restored microbial homeostasis in the liver, promoting the amelioration of liver inflammation. Overall, our findings indicate that DHI-VAP2000@CD represents a promising therapeutic approach for liver fibrosis by specifically targeting HSCs and restoring the liver microbial balance.

Ceriporiopsistianshanensis (Polyporales, Agaricomycetes) and Sideratianshanensis (Hymenochaetales, Agaricomycetes), two new species of wood-inhabiting fungi from Xinjiang, Northwest China.

Wood-inhabiting fungi are abundant in China, but their distribution is uneven, with more fungi in southwest China and fewer fungi in northwest China. During the investigation of wood-inhabiting fungi in Xinjiang, we collected a large number of specimens. Eight specimens growing on Piceaschrenkiana were collected from Tianshan Mountains, and they were described as two new species in Ceriporiopsis and Sidera based on morphological characters and molecular evidence. Ceriporiopsistianshanensis is characterized by a cream to salmon-buff pore surface, larger pores measuring 1-3 per mm, and broadly ellipsoid basidiospores 5-6.5 × 3-4 µm. Sideratianshanensis is characterized by annual to perennial basidiocarps, measuring 15 mm thick, pores 5-7 per mm, cream to rosy buff pore surface, and allantoid basidiospores 3-3.5 × 1-1.4 µm. Detailed illustrations and descriptions of the novel species are provided.

catena-Poly[[diaqua-bis(1H-imidazole-κN(3))cobalt(II)]-µ-2,3,5,6-tetra-chloro-tereph-thal-ato-κ(2)O(1):O(4)].

In the title compound, [Co(C(8)Cl(4)O(4))(C(3)H(4)N(2))(2)(H(2)O)(2)](n), the Co(II) ion displays a distorted octa-hedral coordination geometry with two O atoms from two monodentate tetra-chloro-terephthalate dianions, two N atoms from two imidazole mol-ecules and two O atoms from two water mol-ecules. The Co(II) ions are connected via the tetra-chloro-terephthalate dianions into a chain running along the crystallographic [110] direction. Adjacent chains are linked into a two-dimensional network arranged parallel to (010) by classical N-H⋯O and O-H⋯O hydrogen bonds.

2,3-Bis(thio-phen-2-yl)pyrazine-[2,3-f][1,10]phenanthroline.

The mol-ecule of the title compound, C(22)H(12)N(4)S(2), shows no crystallographic symmetry. The thiophene rings form different dihedral angles [40.15 (9) and 15.43 (10)°] with the pyrazine ring. A strong π-π stacking inter-action occurs between adjacent pyrazine-[2,3-f][1,10]phenanthroline units with an inter-planar distance of 3.4352 (16) Å.

Processable Potassium-Carbon Nanotube Film with a Three-Dimensional Structure for Ultrastable Metallic Potassium Anodes.

K metal holds great promise as the ultimate anode candidate for K-ion batteries because of its high theoretical capacity and low operating potential. However, due to its high viscosity and poor mechanical processability, it remains challenging to manufacture potassium anodes with precise parameters by a simple and executable method. In this work, a high-performance potassium-carbon nanotubes (K@CNTs) composite film electrode with a three-dimensional (3D) skeleton and superior processability is prepared by simply incorporating CNTs into molten potassium. The in situ potassiation reaction between CNTs and molten K formed potassium carbide (KC8) so as to obtain a solid-liquid mixture, which can reduce the surface tension of molten potassium and promote the preparation of the K@CNTs film electrode. The composite electrode can be molded into a variety of shapes and thicknesses in accurate dimensions. The porous, well-conducting CNTs act as a 3D skeleton uniformly distributed in the K metal, providing adequate surface and space to accommodate and attract K metal, thereby inhibiting the growth of the potassium dendrites and the volume expansion upon cycling. As a result, the K@CNTs composite anode exhibits excellent cyclability and rate capability in both symmetric and full cells. The superior processability and excellent electrochemical performance make this composite an ideal anode candidate for commercial applications in potassium metal batteries.