Ultrasensitive PD-L1-Expressing Exosome Immunosensors Based on a Chemiluminescent Nickel-Cobalt Hydroxide Nanoflower for Diagnosis and Classification of Lung Adenocarcinoma.

Programmed death ligand-1 (PD-L1)-expressing exosomes are considered a potential marker for diagnosis and classification of lung adenocarcinoma (LUAD). There is an urgent need to develop highly sensitive and accurate chemiluminescence (CL) immunosensors for the detection of PD-L1-expressing exosomes. Herein, N-(4-aminobutyl)-N-ethylisopropanol-functionalized nickel-cobalt hydroxide (NiCo-DH-AA) with a hollow nanoflower structure as a highly efficient CL nanoprobe was synthesized using gold nanoparticles as a "bridge". The resulting NiCo-DH-AA exhibited a strong and stable CL emission, which was ascribed to the exceptional catalytic capability and large specific surface area of NiCo-DH, along with the capacity of AuNPs to facilitate free radical generation. On this basis, an ultrasensitive sandwich CL immunosensor for the detection of PD-L1-expressing exosomes was constructed by using PD-L1 antibody-modified NiCo-DH-AA as an effective signal probe and rabbit anti-CD63 protein polyclonal antibody-modified carboxylated magnetic bead as a capture platform. The immunosensor demonstrated outstanding analytical performance with a wide detection range of 4.75 × 103-4.75 × 108 particles/mL and a low detection limit of 7.76 × 102 particles/mL, which was over 2 orders of magnitude lower than the reported CL method for detecting PD-L1-expressing exosomes. Importantly, it was able to differentiate well not only between healthy persons and LUAD patients (100% specificity and 87.5% sensitivity) but also between patients with minimally invasive adenocarcinoma and invasive adenocarcinoma (92.3% specificity and 52.6% sensitivity). Therefore, this study not only presents an ultrasensitive and accurate diagnostic method for LUAD but also offers a novel, simple, and noninvasive approach for the classification of LUAD.

BRAF-V600E mutations in plasma and peripheral blood mononuclear cells correlate with prognosis of pediatric Langerhans cell histiocytosis treated with first-line therapy.

BACKGROUND: The clinical relevance of BRAF-V600E alleles in peripheral blood mononuclear cells (PBMCs) and the prognostic impact of the mutants in cell-free (cf) and PBMC DNAs of Langerhans cell histiocytosis (LCH) have not been fully clarified in pediatric LCH. METHODS: We retrospectively determined the levels of BRAF-V600E mutation in paired plasma and PBMC samples at the time of diagnosis of LCH. Subsequently, we performed a separate or combined analysis of the clinical and prognostic impact of the mutants. RESULTS: We assessed BRAF-V600E mutation in peripheral blood from 94 patients of childhood LCH. Our data showed that cfBRAF-V600E was related to young age, multiple-system (MS) disease, involvements of organs with high risk, increased risk of relapse, and worse progression-free survival (PFS) of patients. We also observed that the presence of BRAF-V600E in PBMCs at baseline was significantly associated with MS LCH with risk organ involvement, younger age, and disease progression or relapse. The coexisting of plasma(+)/PBMC(+) identified 36.2% of the patients with the worst outcome, and the hazard ratio was more significant than either of the two alone or neither, indicating that combined analysis of the mutation in plasma and PBMCs was more accurate to predict relapse than evaluation of either one. CONCLUSIONS: Concurrent assessment of BRAF-V600E mutation in plasma and PBMCs significantly impacted the prognosis of children with LCH. Further prospective studies with larger cohorts need to validate the results of this study.

DCDC2 inhibits hepatic stellate cell activation and ameliorates CCl4-induced liver fibrosis by suppressing Wnt/ß-catenin signaling.

Liver fibrosis, as a consequence of chronic liver disease, involves the activation of hepatic stellate cell (HSC) caused by various chronic liver injuries. Emerging evidence suggests that activation of HSC during an inflammatory state can lead to abnormal accumulation of extracellular matrix (ECM). Investigating novel strategies to inhibit HSC activation and proliferation holds significant importance for the treatment of liver fibrosis. As a member of the doublecortin domain-containing family, doublecortin domain containing 2 (DCDC2) mutations can lead to neonatal sclerosing cholangitis, but its involvement in liver fibrosis remains unclear. Therefore, this study aims to elucidate the role of DCDC2 in liver fibrosis. Our findings revealed a reduction in DCDC2 expression in both human fibrotic liver tissues and carbon tetrachloride (CCl4)-induced mouse liver fibrotic tissues. Furthermore, exposure to transforming growth factor beta-1(TGF-ß1) stimulation resulted in a dose- and time-dependent decrease in DCDC2 expression. The overexpression of DCDC2 inhibited the expression of α-smooth muscle actin (α-SMA) and type I collagen alpha 1 (Col1α1), and reduced the activation of HSC stimulated with TGF-ß1. Additionally, we provided evidence that the Wnt/ß-catenin signaling pathway was involved in this process, wherein DCDC2 was observed to inhibit ß-catenin activation, thereby preventing its nuclear translocation. Furthermore, our findings demonstrated that DCDC2 could attenuate the proliferation and epithelial-mesenchymal transition (EMT)-like processes of HSC. In vivo, exogenous DCDC2 could ameliorate CCl4-induced liver fibrosis. In summary, DCDC2 was remarkably downregulated in liver fibrotic tissues of both humans and mice, as well as in TGF-ß1-activated HSC. DCDC2 inhibited the activation of HSC induced by TGF-ß1 in vitro and fibrogenic changes in vivo, suggesting that it is a promising therapeutic target for liver fibrosis and warrants further investigation in clinical practice.

A New Event-Triggered Adaptive Fixed-Time Control Design for Uncertain Nonlinear Systems.

This article investigates the problem of dynamic memory event-triggered (DMET) fixed-time tracking control within time-varying asymmetric constraints for nonaffine nonstrict-feedback uncertain nonlinear systems with unmodeled dynamics and unknown disturbances. The existing dynamic event-triggered control methods cannot handle the nonlinear systems with unmodeled dynamics and nonaffine inputs, which greatly limits the applicability of the strategy. To this end, a novel DMET adaptive fuzzy fixed-time control protocol is constructed based on the idea of command filtered backstepping, in which a new dynamic signal function is established to deal with the unmodeled dynamics and an improved DMET mechanism (DMETM) is designed to solve the problem of nonaffine inputs. It is proved that the newly DMET control strategy ensures the tracking error converges to an arbitrarily small compact set in a fixed time and all the signals of the closed-loop systems are bounded. The effectiveness of the proposed approach is demonstrated by two simulation examples.

Systematic HOIP interactome profiling reveals critical roles of linear ubiquitination in tissue homeostasis.

Linear ubiquitination catalyzed by HOIL-1-interacting protein (HOIP), the key component of the linear ubiquitination assembly complex, plays fundamental roles in tissue homeostasis by executing domain-specific regulatory functions. However, a proteome-wide analysis of the domain-specific interactome of HOIP across tissues is lacking. Here, we present a comprehensive mass spectrometry-based interactome profiling of four HOIP domains in nine mouse tissues. The interaction dataset provides a high-quality HOIP interactome resource with an average of approximately 90 interactors for each bait per tissue. HOIP tissue interactome presents a systematic understanding of linear ubiquitination functions in each tissue and also shows associations of tissue functions to genetic diseases. HOIP domain interactome characterizes a set of previously undefined linear ubiquitinated substrates and elucidates the cross-talk among HOIP domains in physiological and pathological processes. Moreover, we show that linear ubiquitination of Integrin-linked protein kinase (ILK) decreases focal adhesion formation and promotes the detachment of Shigella flexneri-infected cells. Meanwhile, Hoip deficiency decreases the linear ubiquitination of Smad ubiquitination regulatory factor 1 (SMURF1) and enhances its E3 activity, finally causing a reduced bone mass phenotype in mice. Overall, our work expands the knowledge of HOIP-interacting proteins and provides a platform for further discovery of linear ubiquitination functions in tissue homeostasis.

Enhancing fuel characteristics and combustion performance of cellulose-rich straws through CO2-assisted torrefaction.

Cellulose-rich straws of corn and rice were torrefied under carbon dioxide, and the fuel characteristics and combustion performance of the obtained biochar were investigated. A high severity resulted in surface collapse, greater pore volume, elimination of oxygen, elevated calorific value, and improved hydrophobicity in biochar. Following carbon dioxide torrefaction, the cellulose content in solid biochar experienced a slight decrease when the temperature was raised to 220 °C for longer residence durations. At 300 °C, the cellulose content in the biochar was nearly eliminated, while the relative proportion of non-sugar organic matter in corn stover and rice straw increased to 87.40 % and 77.27 %, respectively. The maximum calorific values for biochar from corn and rice straws were 22.38 ± 0.03 MJ/kg and 18.72 ± 0.05 MJ/kg. The comprehensive combustion indexes of rice and corn straw samples decreased to 1.06 × 10-7 and 1.31 × 10-7 after torrefaction at 300 °C, respectively. In addition, the initial decomposition temperatures increased by 38 °C and 45 °C, while the ultimate combustion temperatures rose by 13 °C and 16 °C for corn and rice straws, respectively. These results imply an extended combustion timeframe for the torrefied samples.

TRIM27 elicits protective immunity against tuberculosis by activating TFEB-mediated autophagy flux.

Infectious diseases, such as Mycobacterium tuberculosis (Mtb)-caused tuberculosis (TB), remain a global threat exacerbated by increasing drug resistance. Host-directed therapy (HDT) is a promising strategy for infection treatment through targeting host immunity. However, the limited understanding of the function and regulatory mechanism of host factors involved in immune defense against infections has impeded HDT development. Here, we identify the ubiquitin ligase (E3) TRIM27 (tripartite motif-containing 27) as a host protective factor against Mtb by enhancing host macroautophagy/autophagy flux in an E3 ligase activity-independent manner. Mechanistically, upon Mtb infection, nuclear-localized TRIM27 increases and functions as a transcription activator of TFEB (transcription factor EB). Specifically, TRIM27 binds to the TFEB promoter and the TFEB transcription factor CREB1 (cAMP responsive element binding protein 1), thus enhancing CREB1-TFEB promoter binding affinity and promoting CREB1 transcription activity toward TFEB, eventually inducing autophagy-related gene expression as well as autophagy flux activation to clear the pathogen. Furthermore, TFEB activator 1 can rescue TRIM27 deficiency-caused decreased autophagy-related gene transcription and attenuated autophagy flux, and accordingly suppressed the intracellular survival of Mtb in cell and mouse models. Taken together, our data reveal that TRIM27 is a host defense factor against Mtb, and the TRIM27-CREB1-TFEB axis is a potential HDT-based TB target that can enhance host autophagy flux.Abbreviations: ATG5: autophagy related 5; BMDMs: bone marrow-derived macrophages; CFU: colony-forming unit; ChIP-seq: chromatin immunoprecipitation followed by sequencing; CREB1: cAMP responsive element binding protein 1; CTSB: cathepsin B; E3: ubiquitin ligase; EMSA: electrophoretic mobility shift assay; HC: healthy control; HDT: host-directed therapy; LAMP: lysosomal associated membrane protein; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MCOLN1: mucolipin TPR cation channel 1; Mtb: Mycobacterium tuberculosis; NLS: nuclear localization signal; PBMCs: peripheral blood mononuclear cells; PRKA/PKA: protein kinase cAMP-activated; qRT-PCR: quantitative real-time PCR; RFP: RET finger protein; TB: tuberculosis; TBK1: TANK binding kinase 1; TFEB: transcription factor EB; TRIM: tripartite motif; TSS: transcription start site; ULK1: unc-51 like autophagy activating kinase 1.

Fungicide-tolerant persister formation during cryptococcal pulmonary infection.

Bacterial persisters, a subpopulation of genetically susceptible cells that are normally dormant and tolerant to bactericides, have been studied extensively because of their clinical importance. In comparison, much less is known about the determinants underlying fungicide-tolerant fungal persister formation in vivo. Here, we report that during mouse lung infection, Cryptococcus neoformans forms persisters that are highly tolerant to amphotericin B (AmB), the standard of care for treating cryptococcosis. By exploring stationary-phase indicator molecules and developing single-cell tracking strategies, we show that in the lung, AmB persisters are enriched in cryptococcal cells that abundantly produce stationary-phase molecules. The antioxidant ergothioneine plays a specific and key role in AmB persistence, which is conserved in phylogenetically distant fungi. Furthermore, the antidepressant sertraline (SRT) shows potent activity specifically against cryptococcal AmB persisters. Our results provide evidence for and the determinant of AmB-tolerant persister formation in pulmonary cryptococcosis, which has potential clinical significance.

MRPL21 promotes HCC proliferation through TP53 mutation-induced apoptotic resistance.

BACKGROUND AND AIMS: The specific mechanisms underlying the inhibition of hepatocellular carcinoma (HCC) proliferation and metastasis by mitochondrial apoptosis are not yet fully understood. However, it plays a vital role in suppressing HCC's ability to proliferate and spread. The involvement of MRPL21, a member within the family of mitochondrial ribosomal proteins (MRPs), is well-documented in both cellular apoptosis and energy metabolism. This study aims to explore and unravel the underlying mechanisms through which MRPL21 contributes to mitochondrial apoptosis and resistance against apoptosis in HCC. METHODS: To evaluate the level of MRPL21 expression at the gene and protein expression levels, analysis was performed on human liver samples and blood using techniques for quantification. A knockdown plasmid targeting MRPL21 was constructed to investigate its impact on the growth and apoptosis of hepatocellular carcinoma (HCC). To evaluate the impact of MRPL21 knockdown on hepatocellular carcinoma (HCC) cell proliferation and apoptosis, various assays were performed including CCK-8 assays, flow cytometry analysis, detection of reactive oxygen species (ROS), and assessment of mitochondrial membrane potential (MMP). Furthermore, the role of MRPL21 in TP53 mutation was examined using Nutlin-3. RESULTS: In HCC tissues and blood samples, an upregulation of MRPL21 expression was observed when compared to samples obtained from healthy individuals, and it is correlated with a poor prognosis for HCC. Silencing MRPL21 can effectively suppress Hep3B and HCCLM3 cells proliferation by modulating the mitochondrial membrane potential, it triggers the generation of reactive oxygen species (ROS), thereby leading to G0/G1 cell cycle arrest and initiation of early apoptosis. Furthermore, by inhibiting P53 activity, Nutlin-3 treatment can enhance MRPL21-deficiency-mediated apoptosis in Hep3B and HCCLM3 cells. CONCLUSION: Through its influence on TP53 mutation, MRPL21 promotes HCC proliferation and progression while conferring resistance to apoptosis. These findings suggest that MRPL21 holds promise as a valuable biomarker for the treatment of HCC.

Exploration of Key Genes Combining with Immune Infiltration Level and Tumor Mutational Burden in Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a lethal malignancy due to its heterogeneity and aggressive behavior. Recently, somatic mutations and tumor cell interactions with the surrounding tumor immune microenvironment (TIME) have been reported to participate in HCC carcinogenesis and predict HCC progression. In this study, we aimed to investigate the association between tumor mutational burden (TMB) and TIME in HCC. Additionally, we sought to identify differentially expressed genes (DEGs) associated with HCC prognosis and progression. METHODS: The expression, clinical, and mutational data were downloaded from the cancer genome atlas (TCGA) database. The immune infiltration levels and TMB levels of the HCC samples were estimated and the samples were divided into immune cluster (ICR)-1 and 2 based on immune infiltration score and high and low TMB groups based on TMB score. Thereafter, differential gene expression analysis was conducted to identify the DEGs in the ICR1/2 and high/low TMB groups, and the intersecting DEGs were selected. Thereafter, Cox regression analysis was performed on 89 significant DEGs, among which 19 were associated with prognosis. These 19 DEGs were then used to construct a prognostic model based on their expression levels and regression coefficients. Thereafter, we analyzed the DEGs in mutant and wildtype TP53 HCC samples and identified high BCL10 and TRAF3 expression in the mutant TP53 samples. BCL10 and TRAF3 expression was detected by real-time quantitative reverse transcription PCR and immunohistochemistry, and their clinical correlation, biological function, and immune infiltration levels were analyzed by chi-square analyses, Gene Set Enrichment Analysis (GSEA), and "ssGSEA", respectively. RESULTS: The results of our study revealed that immune infiltration level was correlated with TMB and that they synergistically predicted poor prognosis of HCC patients. DEGs enriched in immune-related pathways could serve as indicators of immunotherapy response in HCC. Among these DEGs, BCL10 and TRAF3 were highly expressed in HCC tissues, especially in the mutant TP53 group, and they co-operatively exhibited immunological function, thereby affecting HCC progression and prognosis. CONCLUSION: In this study, we identified BCL10 and TRAF3 as potential prognostic indicators in HCC patients. Additionally, we found that BCL10 and TRAF3 influence TMB and TIME in HCC patients and can be used for the development of immune-based therapies for improving the long-term survival of HCC patients.

Efficacy and safety of agomelatine in epilepsy patients with sleep and mood disorders: An observational, retrospective cohort study.

OBJECTIVE: To evaluate the therapeutic efficacy and safety of agomelatine for treating the sleep and mood disorders in epilepsy patients. METHODS: Retrospective data were derived from 113 epilepsy patients for at least 8 weeks. All the subjects were divided into two groups, one was treated with agomelatine, the other was treated with escitalopram. Their depression and anxiety states were assessed by Hamilton Depression (HAMD) and Hamilton Anxiety (HAMA) Scales. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI). RESULTS: The HAMA, HAMD and PSQI scores in both groups significantly declined after the treatments with agomelatine and escitalopram. However, the agomelatine group exhibited greater improvement in terms of HAMA and PSQI scores compared to the escitalopram group. No severe adverse events were observed in agomelatine group. SIGNIFICANCE: Agomelatine performed better in HAMA and PSQI scores compared to escitalopram, where no significant increase in seizure frequency or side effects were observed. Possibly, agomelatine presents a promising therapeutic option for treating the sleep or mood disorders in epilepsy patients.

Tai Chi Effects on Heart Rate Variability: A Systematic Review and Meta-Analysis.

Background: Tai Chi, originating in China, is a mind-body exercise that integrates mind training, Qi training, and body training. While Tai Chi practice has demonstrated beneficial physical and psychological health benefits, how it achieves effects has been less studied. In recent years, some researchers have begun to explore the potential of heart rate variability (HRV) changes as balancing the autonomic nervous system (ANS) to achieve results, but only a few studies have directly tested how Tai Chi impacts HRV, and results have been mixed. Objective: This study systematically evaluates the effects of Tai Chi on HRV. Methods: Electronic databases including Web of Science, PubMed, Scopus, Cochrane Library, and EBSCOhost direct from inception until December 2020 were searched to obtain eligible studies. The primary outcomes examined were HRV parameters (time domain and frequency domain), and secondary analysis was whether breathing was emphasized during Tai Chi interventions. Results: Based on the 11 randomized controlled trials included comparing Tai Chi with non-active control condition, meta-analyses showed significantly beneficial effects on HRV parameters (low-frequency power [LF], mean difference [MD] = -200.40, 95% confidence interval [CI]: 365.31 to -35.49, p = 0.02; normalized low-frequency power [nLF], MD = -7.39, 95% CI: -12.48 to -2.29, p = 0.004; total power [TP], MD = -649.82, 95% CI: -1180.54 to -119.11, p = 0.02; very low-frequency power [VLF], MD = -200.55, 95% CI: -349.63 to -51.46, p = 0.008; root mean square of the square sum of the difference between adjacent normal heartbeat intervals [RMSSD], MD = 2.59, 95% CI: -7.23 to 12.41, p < 0.0001; standard deviation of the heartbeat interval [SDNN], MD = 8.33, 95% CI: 0.69 to 15.98, p = 0.03). The effect of Tai Chi on certain HRV parameters compared with active control conditions was less favorable (nLF, MD = 6.42, 95% CI: 3.74 to 9.09, p < 0.00001; normalized high-frequency power [nHF], MD: -6.13, 95% CI: -8.76 to -3.50, p < 0.00001; low-frequency/high-frequency power ratio [LF/HF], MD = 1.00, 95% CI: 0.68 to 1.33, p < 0.00001). In addition, studies describing an emphasis on breathing in the Tai Chi intervention showed positive impact on HRV (nLF, MD = -3.22, 95% CI: -5.32 to -1.12, p = 0.003; nHF, MD = 3.80, 95% CI: 0.80 to 6.81, p = 0.01; SDNN, MD = 5.55, 95% CI: 4.85 to 6.25, p < 0.00001). Conclusions: Meta-analysis results showed that Tai Chi compared with non-active control conditions has a positive impact on key HRV parameters associated with the ANS balance. However, active control condition comparisons showed greater HRV improvements than for Tai Chi. Considering the limited number of studies and their heterogeneity, further robust studies are necessary to verify findings and investigate differential effects of Tai Chi on HRV.

The relationship between incidence of cesarean section and physical activity during pregnancy among pregnant women of diverse age groups: Dose-response meta-analysis.

BACKGROUND: There is a lack of evidence related to physical activity and risk of cesarean section (CS) by age and/or weight in pregnant women. OBJECTIVES: To evaluate the effect of physical activity on the incidence of CS and explore the relationship of age and body mass index (BMI) with incidence of CS. SEARCH STRATEGY: A systematic search was conducted in CNKI, WANGFANG, Web of Science, and PubMed from inception to 31 August 2021. SELECTION CRITERIA: Experimental studies were included if the participants were pregnant, if intervention included physical activity and controls received routine prenatal care only, and if primary outcome was CS. DATA COLLECTION AND ANALYSIS: Meta-analysis included a heterogeneity test, data combination, subgroup analysis, forest plot, sensitivity analysis, and dose-response regression analysis. MAIN RESULTS: Sixty-two studies were included. Physical activity during pregnancy decreased the incidence of CS (relative risk [RR] 0.81, 95% confidence interval [CI] 0.74-0.88, P < 0.001). The incidence of CS was lower among the overweight/obese group (RR 0.78, 95% CI 0.65-0.93) compared with the normal weight group (RR 0.82, 95% CI 0.74-0.90). The incidence of CS was lowest among the young age group (RR 0.61, 95% CI 0.46-0.80) compared with the middle age group (RR 0.74, 95% CI 0.64-0.85) and the older age group (RR 0.90, 95% CI 0.82-1.00). The critical value, when age becomes a risk factor for CS, was 31.7 years in the intervention group and 28.5 years in the control group. CONCLUSIONS: Physical activity during pregnancy can reduce the incidence of CS, especially among obese people, and prolong the gestational age span.

Family caregiver's willingness to care from the perspective of altruism.

Objectives: The willingness of family members to take care of older relatives directly affects the quality of life of disabled older adults, so it is necessary to understand the status quo of willingness to care and its influencing factors. This has been extensively studied in other countries, but, it is rarely studied in China. Based on the theory of altruism, employing a unique sample from Shanghai, China in 2017 and 2022, we attempt to reveal the influencing factors of the care willingness of family caregivers during the transition period. Methods: To measure caregiver burden and functional disability of the care recipient, we employ the Zarit Burden Interview (ZBI) and the Barthel Index, respectively. Then we utilized the ordinary least squares (OLS) methodology and estimated four regression models. Models 1, 2, and 3 examined the impact of the variables of the caregiver burden, responsibility and love, and the quality of the caregiver-caregiver recipient relationship, respectively, on family caregivers' willingness to care. Model 4 was the full model. To testify whether the caregiver burden is likely to act as a mediator, path analysis was used, and the path was adjusted and verified. Results: According to the survey, in Shanghai, only half of the caregivers had a very high care willingness to care for disabled older relatives, while nearly one-tenth of the caregivers had a low willingness. It was the caregiver burden rather than the functional disability of older adults that harms family caregivers' willingness to care. Responsibility and caring out of love were positively related to care willingness. Relationship quality was the most important influencing factor, explaining 10.2% of the variance in care willingness. Path analysis demonstrated that responsibility, caring out of love, and relationship quality directly and through the mediation of caregiver burden indirectly affected care willingness. Conclusion: Our results revealed that reciprocal altruism presented by the quality of the caregiver-care recipient relationship had a significantly positive impact on family caregivers' willingness to care. In addition, the caregiver burden was found not only directly affected care willingness, but also acted as a mediator. To promote the perfection of laws and policies, comprehensive samples of different types of cities should be included and the measurement of key variables could be further improved in future studies.

Synthesis and Fluorescence Sensing for Nitro Explosives and Pesticides of Two Cd-Coordination Polymers.

Two cadmium coordination polymers (CPs), {[Cd(zgt)(2,2'-bipy)(H2O)]·H2O}n (1) and {[Cd(zgt)(BPP)(H2O)]·H2O}n (2) (H2zgt = 5-methoxyresorcinic acid, 2,2'-bipy = 2,2'-bipyridine, and BPP = 1,3-bis(4-pyridyl)propane), were prepared by the hydrothermal method. The structures of CPs 1-2 were characterized by IR, TGA, X-ray powder diffraction, and elemental analysis. The single-crystal structure analysis shows that CP 1 is a typical 1D chain structure and CP 2 belongs to a 2D layered structure. Based on the excellent luminescence properties of CP 1 and 2, fluorescence sensing experiments were carried out for explosives and pesticides. The results of the explosion sensing experiment showed that CP 1 and 2 had an excellent fluorescence quenching effect on PNBA (p-nitrobenzoic acid) and TNP (2,4,6-trinitrophenol), respectively, and the detection limits were 3.28 and 11.4 nM, respectively. Interestingly, both CP 1 and 2 showed good fluorescence quenching against the pesticide fluridine (Flu), and CP 1 had a lower detection limit and was more sensitive. In addition, the fluorescence quenching mechanism was discussed in detail by the UV absorption spectrum and density functional theory. In order to explore its practical application, the content of Flu in water samples was detected by a labeling recovery method.

[Quality evaluation of Lysimachiae Herba from different habitats based on simultaneous determination of multiple bioactive constituents combined with multivariate statistical analysis].

A method based on ultra-high performance liquid chromatography coupled with triple quadrupole linear ion trap-tandem mass spectrometry(UHPLC-QTRAP-MS/MS) was developed for the simultaneous determination of 41 bioactive constituents of flavonoids, organic acids, nucleosides, and amino acids in Lysimachiae Herba. The content of multiple bioactive constituents was compared among the samples from different habitats. The chromatographic separation was performed in a Waters XBridge®C_(18) column(4.6 mm×100 mm, 3.5 µm) at 30 ℃. The gradient elution was performed with 0.4% methanol(A)-formic acid water(B) as the mobile phase at a flow rate of 0.8 mL·min~(-1), and the multiple-reaction monitoring(MRM) mode was adopted. According to the content of 41 constituents, hierarchical cluster analysis(HCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and gray relational analysis(GRA) were perfomed to comprehensively evaluate the samples from different habitats. The results showed that the 41 constituents exhibited good linear relationship within the tested concentration ranges, with the correlation coefficients(r) greater than 0.999 4. The method featured good precision, repeatability, and stability with the relative standard deviations(RSDs) less than 5.0%. The average recoveries of the 41 constituents ranged from 98.06% to 101.9%, with the RSDs of 0.62%-4.6%. HCA and OPLS-DA separated 48 batches of Lysimachiae Herba samples from different habitats into three categories: the producing areas in Sichuan and Chongqing, the producing areas in Jiangsu, Zhejiang, and Jiangxi, and the producing areas in Guizhou. The content of 41 constituents varied among the Lysimachiae Herba samples from different habitats. The GRA results revealed that the Lysimachiae Herba sample from Nanchong City, Sichuan Province had the best comprehensive quality. The method developed in this study was accurate and reliable and thus can be used for comprehensive evaluation of Lysimachiae Herba quality and provide basic information for the selection of habitats.

A self-enhanced electrochemiluminescence array chip for portable label-free detection of SARS-CoV-2 nucleocapsid protein with smartphone.

SARS-CoV-2 antigen detection plays a key role in the rapid diagnosis of COVID-19. However, current clinically used immunoassays are often limited by assay throughput, sensitivity, accuracy, and field operating conditions. To address these challenges, we constructed a self-enhanced electrochemiluminescence (ECL) array chip (SE2AC) for highly sensitive and label-free detection of SARS-CoV-2 nucleocapsid protein (N protein) with a facile and portable assay setup. Firstly, the self-enhanced ECL nanomaterials with inherent film-forming properties were synthesized by co-doping Ru(bpy)32+ and polyethyleneimine (PEI) in silica nanoparticles (Ru/PEI@SiO2). Secondly, a resistance-induced potential difference-based single-electrode electrochemical system (SEES) was adapted to serve as the electrode array to facilitate one-step assembly without the need for chip alignment. Thirdly, the chip electrode array was functionalized with the synthesized self-enhanced ECL emitters and captured antibodies. In addition, a portable detection box equipped with a smartphone was 3D-printed to serve as the chip holder and "dark room" for imaging acquisition. The SE2AC performance was validated with N protein with a limit of detection (LOD) of 0.47 pg/mL in the range of 1-10,000 pg/mL. Furthermore, the chip successfully detected the viral antigen residue as low as 1.92 pg/mL from diluted rehabilitation patients' serum samples. This is the first study reporting label-free detection of SARS-Cov-2 N protein based on a self-enhanced ECL immunosensor, which provides a novel facile method for highly sensitive diagnosis of COVID-19 with high throughput, portability, and low cost.

Synthesis of lignin nanoparticle­manganese dioxide complex and its adsorption of methyl orange.

Lignin nanoparticles (LNPs) were synthesized using an anti-solvent method and subsequently loaded with manganese dioxide (MnO2) via potassium permanganate treatment, resulting in the formation of MnO2@LNPs. An extensive investigation was conducted to elucidate the influence of MnO2@LNPs on the decolorization of methyl orange solution. The LNPs were successfully obtained by adjusting the preparation parameters, yielding particles exhibited average sizes ranging from 300 to 600 nm, and the synthesis process exhibited a high yield of up to 87.3% and excellent dispersion characteristics. Notably, LNPs size was reduced by decreasing initial concentration, increasing stirring rate, and adding water. In the acetone-water two-phase system, LNPs self-assembled into spherical particles driven by π-π interactions and hydrogen bond forces. Oxidation modification using potassium permanganate led to the formation of nanoscale MnO2, which effectively combined with LNPs. Remarkably, the resulting MnO2@LNPs demonstrated a two-fold increase in methyl orange adsorption capacity (227 mg/g) compared to unmodified LNPs. The process followed the Langmuir isotherm model and was exothermic.

[COL4A5 genotypes and clinical characteristics of children with Alport syndrome]. / 儿童Alport综合征COL4A5åŸºå› åž‹åŠä¸´åºŠç‰¹ç‚¹åˆ†æž.

OBJECTIVES: To investigate the genotypes of the pathogenic gene COL4A5 and the characteristics of clinical phenotypes in children with Alport syndrome (AS). METHODS: A retrospective analysis was performed for the genetic testing results and clinical data of 19 AS children with COL4A5 gene mutations. RESULTS: Among the 19 children with AS caused by COL4A5 gene mutations, 1 (5%) carried a new mutation of the COL4A5 gene, i.e., c.3372A>G(p.P1124=) and presented with AS coexisting with IgA vasculitis nephritis; 3 children (16%) had large fragment deletion of the COL4A5 gene, among whom 2 children (case 7 had a new mutation site of loss51-53) had gross hematuria and albuminuria at the onset, and 1 child (case 13 had a new mutation site of loss3-53) only had microscopic hematuria, while the other 15 children (79%) had common clinical phenotypes of AS, among whom 7 carried new mutations of the COL4A5 gene. Among all 19 children, 3 children (16%) who carried COL4A5 gene mutations also had COL4A4 gene mutations, and 1 child (5%) had COL4A3 gene mutations. Among these children with double gene mutations, 2 had gross hematuria and proteinuria at the onset. CONCLUSIONS: This study expands the genotype and phenotype spectrums of the pathogenic gene COL4A5 for AS. Children with large fragment deletion of the COL4A5 gene or double gene mutations of COL4A5 with COL4A3 or COL4A4 tend to have more serious clinical manifestations.

N-(4-Aminobutyl)-N-ethylisopropanol and Co2+ Dual-Functionalized Core-Shell Fe3O4@Au/Ag Magnetic Nanomaterials with Strong and Stable Chemiluminescence for a Label-Free Exosome Immunosensor.

Recently, magnetic beads (MBs) are moving toward chemiluminescence (CL) functional magnetic nanomaterials with a great potential for constructing label-free immunosensors. However, most of the CL-functionalized MBs suffer from scarce binding sites, easy aggregation, and leakage of CL reagents, which will ultimately affect the analytical performance of immunosensors. Herein, by using core-shell Fe3O4@Au/Ag magnetic nanomaterials as a nanoplatform, a novel N-(4-aminobutyl)-N-ethylisopropanol (ABEI) and Co2+ dual-functionalized magnetic nanomaterial, namely, Fe3O4@Au/Ag/ABEI/Co2+, with strong and stable CL emission was successfully synthesized. Its CL intensity was 36 and 3.5 times higher than that of MB@ABEI-Au/Co2+ and ABEI and Co2+ dual-functionalized chemiluminescent MBs previously reported by our group, respectively. It was found that the excellent CL performance of Fe3O4@Au/Ag/ABEI/Co2+ could be attributed to the enrichment effect of the Au/Ag shell and the synergistic enhance effect of the Au/Ag shell and Co2+. A related CL mechanism has been proposed. Afterward, based on the intense and stable CL emission of Fe3O4@Au/Ag/ABEI/Co2+, a sensitive and effective label-free CL immunosensor for exosome detection was established. It exhibited excellent analytical performance with a wide detection range of 3.1 × 103 to 3.1 × 108 particles/mL and a low detection limit of 2.1 × 103 particles/mL, which were better than the vast majority of the reported CL immunosensors. Moreover, the proposed label-free CL immunosensor was successfully used to detect exosomes in human serum samples and enabled us to distinguish healthy persons and lung cancer patients. It has the potential to be a powerful tool for exosome study and early cancer diagnosis.