Diagnosing, Typing, and Staging of Renal Cell Carcinoma by Designer Matrix-Based Urinary Metabolic Analysis.

Molecular diagnosing, typing, and staging have been considered to be the ideal alternatives of imaging-based detection methods in clinics. Designer matrix-based analytical tools, with high speed, throughout, efficiency and low/noninvasiveness, have attracted much attention recently for in vitro metabolite detection. Herein, we develop an advanced metabolic analysis tool based on highly porous metal oxides derived from available metal-organic frameworks (MOFs), which elaborately inherit the morphology and porosity of MOFs and newly incorporate laser adsorption capacity of metal oxides. Through optimized conditions, direct high-quality fingerprinting spectra in 0.5 µL of urine are acquired. Using these fingerprinting spectra, we can discriminate the renal cell carcinoma (RCC) from healthy controls with higher than 0.99 of area under the curve (AUC) values (R2Y(cum) = 0.744, Q2 (cum) = 0.880), as well, from patients with other tumors (R2Y(cum) = 0.748, Q2(cum) = 0.871). We also realize the typing of three RCC subtypes, including clear cell RCC, chromophobe RCC (R2Y(cum) = 0.620, Q2(cum) = 0.656), and the staging of RCC (R2Y(cum) = 0.755, Q2(cum) = 0.857). Moreover, the tumor sizes (threshold value is 3 cm) can be remarkably recognized by this advanced metabolic analysis tool (R2Y(cum) = 0.710, Q2(cum) = 0.787). Our work brings a bright prospect for designer matrix-based analytical tools in disease diagnosis, typing and staging.

In Vitro Diagnostic Examination and Prognosis Surveillance by Hierarchical Heterojunction-Assisted Metabolic Analysis.

High-throughput metabolic analysis based on laser desorption/ionization mass spectrometry exhibits broad prospects in the field of large-scale precise medicine, for which the assisted ionization ability of the matrix becomes a determining step. In this work, the gold-decorated hierarchical metal oxide heterojunctions (dubbed Au/HMOHs) are proposed as a matrix for extracting urine metabolic fingerprints (UMFs) of primary nephrotic syndrome (PNS). The hierarchical heterojunctions are simply derived from metal-organic framework (MOF)-on-MOF hybrids, and the native built-in electric field from heterojunctions plus the extra Au decoration provides remarkable ionization efficiency, attaining high-quality UMFs. These UMFs are employed to realize precise diagnosis, subtype classification, and effective prognosis evaluation of PNS by appropriate machine learning, all with 100% accurate ratios. Moreover, a high-confidence marker panel for PNS diagnosis is constructed. Interestingly, all panel metabolite markers present obviously uniform downregulation in PNS compared to healthy controls, shedding light on mechanism exploration and pathway analysis. This work drives the application of metabolomics toward precision medicine.

Recognition of urinary N-linked glycopeptides in kidney cancer patients by hydrophilic carbohydrate functionalized magnetic metal organic framework combined with LC-MS/MS.

A hydrophilic carbohydrate functionalized magnetic metal organic framework (Mag Zr-MOF@G6P) was synthesized via a facile one-step modification strategy for selective glycopeptide capture in virtue of hydrophilic interaction chromatography technique. The inherently hydrophilic Zr-MOF layer not only provides selective size-sieving pore structures but also offers large specific surface area to afford abundant affinity sites. Hydroxyl-rich glucose-6-phosphate was immobilized onto the Zr-MOF via a straightforward coordination manner to regulate its surface property, for the purpose of enhancing its hydrophilicity. Benefitting from the merits of Zr-MOF and glucose-6-phosphate, the as-designed composite exhibits good selectivity (the mass ratio of HRP digests to BSA digests was up to1:200) and low limit of detection (0.1 fmol µL-1) towards the recognition of glycopeptides from standard samples. More excitingly, glycopeptides in urine of healthy people and patients with kidney cancer were successfully enriched and identified by the combined liquid chromatography-mass spectrometry/mass spectrometry technology (LC-MS/MS). Further gene ontology analysis of molecular function and biological process revealed that 13 original glycoproteins of the identified glycopeptides from urine of patients significantly participate in diverse cancer-associated events, including collagen binding, immunoglobulin receptor binding, antigen binding, and complement activation process. Graphical abstract.

Dual metal cations coated magnetic mesoporous silica probe for highly selective capture of endogenous phosphopeptides in biological samples.

For the first time, dual metal ions (Ti4+-Zr4+) were successfully modified into the channel of magnetic mesoporous silica to obtain an affinity probe for highly selective capture of endogenous phosphopeptides in biological samples. The newly prepared Fe3O4@mSiO2@Ti4+-Zr4+ composites possessed the advantages of ordered mesoporous channels, superparamagnetism, and enhanced affinity properties of dual metal ions of Ti4+ and Zr4+. The phosphopeptide enrichment efficiency of the Fe3O4@mSiO2@Ti4+-Zr4+ composite was investigated, and the result indicated an ultrahigh size exclusive ability (weight ratio of ß-casein tryptic digests, BSA, and α-casein protein reached up to 1:1000:1000). Compared to magnetic affinity probes with single metal ions (Fe3O4@mSiO2@Ti4+, Fe3O4@mSiO2@Zr4+), the composite possessed stronger specificity, higher sensitivity, and better efficiency; and more importantly, it showed much enhanced enrichment ability towards both mono- and multi-phosphorylated peptides. Additionally, by utilizing the Fe3O4@mSiO2@Ti4+-Zr4+ affinity probe, a total number of 104 endogenous phosphopeptides including 95 mono-phosphopeptides and 9 multi-phosphopeptides were captured and identified from human saliva, indicating the great potential for the application of the novel probe for the peptidome analysis in the future. Graphic abstract.

Enhancing rural B&B management through machine learning and evolutionary game: A case study of rural revitalization in Yunnan, China.

The rural B&B industry is a key component of rural tourism, local economic development, and the wider rural revitalization strategy. Despite the abundance of tourism resources in Yunnan, the B&B sector faces significant challenges. It is therefore imperative to accurately identify the most pressing issues within the current B&B industry and formulate appropriate solutions to advance Yunnan's rural revitalization efforts. This study uses recent reviews of rural B&Bs on Ctrip.com and employs machine learning techniques, including Bert, CNN, LSTM, and GRU, to identify the key management challenges currently facing Yunnan's rural B&B industry. An analysis is then conducted to identify the key stakeholders involved in the process of improving the management of Yunnan's B&Bs. To assess the willingness of each stakeholder to support the improvement of the rural B&B industry, this paper establishes a three-party evolutionary game model and examines the dynamic evolutionary process of management improvement within Yunnan's rural B&B industry. Two scenarios of evolutionarily stable strategies are analyzed, and parameters impacting stakeholders' strategy choices are simulated and evaluated. The results show that: i) Improving the "human factor" is the top priority for the current management improvement because tourists are most concerned about the emotional experience. Operators need to focus on improving service attitude and emotional experience; ii) The main stakeholders in the current management optimization process of Yunnan B&Bs are the local government, B&B operators, and tourists. Under appropriate conditions, the evolutionarily stable strategy of (1, 1, 1) is reachable. iii) variables such as additional costs, tourists' choice preferences, and government penalties significantly affect the strategy choices of stakeholders, especially B&B operators. This paper offers effective strategies for improving B&B management that can benefit the government, B&B operators, and tourists, and ultimately contribute to the promotion of quality rural revitalization. The paper not only identifies focal areas for improving B&B management in rural Yunnan, but also provides an in-depth understanding of stakeholder dynamics. As a result, it provides valuable insights to further the cause of quality rural revitalization.

Aptamer-aided plasmonic nano-urchins for reporter-free surface-enhanced Raman spectroscopy analysis of cortisol.

Cortisol is a vital glucocorticoid hormone reflecting stress levels and related disease processes. In this study, we report an aptamer-functionalized plasmonic nano-urchin (α-FeOOH@Au-aptamer)-aided cortisol-capturing and surface-enhanced Raman spectroscopy (SERS) analysis approach. The designed α-FeOOH@Au-aptamer exhibits a well-patterned plasma structure, which combines the good SERS enhancement ability of reduced nanogaps between the Au plasma and the hot spot-favored structure of anisotropic tips from α-FeOOH urchins, with the high affinity of the aptamer towards cortisol molecules. The α-FeOOH@Au-aptamer achieved reporter-free SERS quantification for cortisol with good sensitivity (limit of detection <0.28 µmol L-1), robust salt (1.0 mol per L NaCl) and protein (5.0 mg per mL bovine serum protein) tolerance, favorable reproducibility, as well as good reusability. We further demonstrated the good cortisol-capturing ability and SERS efficacy of the α-FeOOH@Au-aptamer profiling in the serum and urine samples. Our approach provides an alternative tool for cortisol analysis and a reference strategy for report-free SERS detection of small molecules.

Hollow Crystallization COF Capsuled MOF Hybrids Depict Serum Metabolic Profiling for Precise Early Diagnosis and Risk Stratification of Acute Coronary Syndrome.

Acute coronary syndrome (ACS), comprising unstable angina (UA) and acute myocardial infarction (AMI), is the leading cause of death worldwide. Currently, lacking effective strategies for classifying ACS hinders the prognosis improvement of ACS patients. Disclosing the nature of metabolic disorders holds the potential to reflect disease progress and high-throughput mass spectrometry-based metabolic analysis is a promising tool for large-scale screening. Herein, a hollow crystallization COF capsuled MOF hybrids (UiO-66@HCOF) assisted serum metabolic analysis is developed for the early diagnosis and risk stratification of ACS. UiO-66@HCOF exhibits unrivaled chemical and structural stability as well as endowing satisfying desorption/ionization efficiency in the detection of metabolites. Paired with machine learning algorithms, early diagnosis of ACS is achieved with the area under the curve (AUC) value of 0.945 for validation sets. Besides, a comprehensive ACS risk stratification method is established, and the AUC value for the discrimination of ACS from healthy controls, and AMI from UA are 0.890, and 0.928. Moreover, the AUC value of the subtyping of AMI is 0.964. Finally, the potential biomarkers exhibit high sensitivity and specificity. This study makes metabolic molecular diagnosis a reality and provided new insight into the progress of ACS.

Metabolic Molecular Diagnosis of Inflammatory Bowel Disease by Synergistical Promotion of Layered Titania Nanosheets with Graphitized Carbon.

Due to inefficient diagnostic methods, inflammatory bowel disease (IBD) normally progresses into severe complications including cancer. Highly efficient extraction and identification of metabolic fingerprints are of significance for disease surveillance. In this work, we synthesized a layered titania nanosheet doped with graphitized carbon (2D-GC-mTNS) through a simple one-step assembly process for assisting laser desorption ionization mass spectrometry (LDI-MS) for metabolite analysis. Based on the synergistic effect of graphitized carbon and mesoporous titania, 2D-GC-mTNS exhibits good extraction ability including high sensitivity (< 1 fmol µL-1) and great repeatability toward metabolites. A total of 996 fingerprint spectra were collected from hundreds of native urine samples (including IBD patients and healthy controls), each of which contained 1220 m/z metabolite features. Diagnostic model was further established for precise discrimination of patients from healthy controls, with high area under the curve value of 0.972 and 0.981 toward discovery cohort and validation cohort, respectively. The 2D-GC-mTNS promotes LDI-MS to be close to clinical application, with rapid speed, minimum sample consumption and free of sample pretreatment. Supplementary Information: The online version contains supplementary material available at 10.1007/s43657-022-00055-0.

Specific enrichment and glycosylation discrepancy profiling of cellular exosomes using a dual-affinity probe.

A hydrophilic probe is employed to enrich exosomes from three kinds of cancer cells by TiO2-phosphate interaction and exosomal glycoproteins by hydrophilic interaction in succession. The probe performs efficiently in both the enrichment processes. And the analytical results confirm that unique exosomal glycoproteins can distinguish parent exosomes from others.

One-pot preparation of hydrophilic citric acid-magnetic nanoparticles for identification of glycopeptides in human saliva.

A facile and mild approach was carried out to synthesize citrate acid-magnetic ferroferric oxide for glycopeptide analysis. The material was synthesized successfully and applied in glycopeptide identification from human saliva, indicating that this method could be a promising tool for glycopeptidome analysis, which also enlightened the simple fabrication of hydrophilic materials in analytical science.

Magnetic mesoporous silica of loading copper metal ions for enrichment and LC-MS/MS analysis of salivary endogenous peptides.

Peptidomics research is of great significance for discovering potential biomarkers and monitoring human diseases. As a kind of common clinical biofluid, saliva known for its noninvasive collection and easy accessibility has been widely used in peptidomics research. In this article, we combined immobilized metal ions affinity chromotography (IMAC) with mesoporous material and proposed the copper ion doped magnetic mesoporous silica material (denoted as Fe3O4@mSiO2-Cu2+) which had a large surface area of 221 m2 g-1 and pore volume of 0.20 cm3 g-1. By immobilizing copper ions onto the mesopore walls, the standard peptide Angiotensin II could be identified in an extremely low concentration of 0.1 fmol µl-1 and in a mass ratio of 1:500 (Angiotensin II:BSA, m/m), which indicated significant sensitivity and a great size-exclusive ability. In addition, the introduction of polydopamine (PDA) made Fe3O4@mSiO2-Cu2+ more hydrophilic and biocompatible which could improve the profiling of endogenous peptides in bio-sample. Finally, 131 endogenous peptides were identified in human saliva after enrichment with Fe3O4@mSiO2-Cu2+. Therefore, Fe3O4@mSiO2-Cu2+ nanoparticles provided a promising candidate protocol for biomarker discovery.

Aptamer-functionalized magnetic metal organic framework as nanoprobe for biomarkers in human serum.

Human serum is a huge bioinformatics database of human physiological and pathological state, many proteins/peptides among which can serve as biomarkers for monitoring human's health condition, thereby being worth exploring. The simple and fast capture of biomarkers from human serum is the first key step to realize their accurate detection. In this work, we developed the aptamer functionalized magnetic metal organic framework nanoprobe, and furtherly combined with mass spectrometry technology to establish an efficient method of identifying biomarkers. Taking insulin as example of biomarker in human serum, we developed sulfhydryl human insulin aptamer functionalized magnetic metal organic framework (denoted as Mag MOF@Au@HIA) through the post-synthetic modification of MIL-101(Cr)-NH2 for testing the applicability of the established method. Depending on the strong magnetic responsiveness and high specific area as well as high-loaded human insulin aptamers, the limit of detection of insulin was down to 1 ng/mL and 2 ng/mL in the standard insulin solution and human serum, respectively. Moreover, a good linear relationship (R2 = 0.998) was obtained by using standard insulin solution with concentration range from 100 ng/mL to 5 ng/mL, based on which the capture recovery of insulin with Mag MOF@Au@HIA from human serum was demonstrated to be excellent. All of the results indicate that the aptamer-functionalized magnetic metal organic framework is a promising nanoprobe for biomarkers capture in human serum.

Magnetic metal-organic frameworks containing abundant carboxylic groups for highly effective enrichment of glycopeptides in breast cancer serum.

A mercaptosuccinic acid functionalized hydrophilic magnetic metal-organic framework nanocomposite (denoted as mMOF@Au-MSA) was proposed and synthesized to provide an excellent platform for glycopeptide analysis. The novel nanomaterial integrated favorable advantages such as robust magnetic response from Fe3O4 magnetic nanoparticles, large surface area contributed by MOF, abundant ultra-high hydrophilic carboxylic groups from mercaptosuccinic acid, as well as unbiased affinity toward different types of glycopeptides. This nanocomposite was successfully utilized to capture glycopeptides from standard protein digests with the high selectivity and great sensitivity of 0.5 fmol µL-1. Notably, 307 glycopeptides assigned to 96 glycoproteins were identified from only 2 µL serum of breast cancer patient. The satisfying achievement indicated that the as-prepared nanopartical had promising potential in exploring the knowledge of glycoproteins in breast cancer.

[Separation and determination of optical isomers of phenylephrine by chiral ligand exchange capillary elcctrophoresis coupling with the promoting effect of ionic liquid].

A method for the separation and determination of optical isomers of phenylephrine was developed based on the promoting effect of non-chiral ionic liquid on chiral ligand-exchange capillary electrophoresis after the electrophoretic parameters were optimized systematically. R-phenylephrine and S-phenylephrine can be separated and determined effectively in 20 mmol/L Tris-H3PO4 buffer solution (pH 5.4) composed of 4.0 mmol/L Cu(II), 8.0 mmol/L L-proline (L-Pro) and 15 mmol/L 1-butyl-3-methylimidazolium chloride ([BMIM] Cl) with the applied voltage of 20 kV, capillary temperature of 25 °C , detection wavelength of 254 nm, and injection of 5 s at 3,447 Pa. The resolution of R- and S-phenylephrines was 1. 42. The linear ranges for the determination of R-phenylephrine and S-phenylephrine were 12. 5 - 150 mg/L and 15. 0-150 mg/L, respectively. The method has been satisfactorily used for the determination of R-phenylephrine and S-phenylephrine in the spiked blood and urine samples. The spiked recoveries in the urine sample were in the range of 93. 7% -108. 2% with the RSDs lower than 3. 18% (n= 3) , and the spiked recoveries in the blood sample were in the range of 91. 4% and 113. 1% with the RSDs lower than 4. 82% (n =3).

[Determination of loureirin A and loureirin B in dragon's blood by capillary zone electrophoresis].

A capillary zone electrophoresis method (CZE) for the simultaneous determination of loureirin A and loureirin B was developed based on the optimized conditions of the pH, composition and concentration of the running buffer solution. Loureirin A and loureirin B were separated and determined effectively within 15 min in a running buffer solution of 20 mmol/L Na2B4O7 (pH 9.98 adjusted with NaOH solution) containing 10.0% (v/v) acetonitrile, 5.0% (v/v) ethylene glycol and 1.0% (v/v) butanol, with the applied voltage of 20 kV, capillary temperature of 25 degrees C, detection wavelength of 211 nm, and injection of 5 s at 3447 Pa. The linear ranges for the determination of loureirin A and loureirin B were 1.00-100 mg/L and 0.50-100 mg/L, respectively. The determination of loureirin A and loureirin B in dragon's blood from natural and artificial inoculation was performed by the proposed method. The relative standard deviations for the determination of the two constituents in samples were from 0.6% to 3.8%, and the recoveries ranged between 95.1% and 105.8%. The method is simple, rapid and possesses higher reproducibility and efficiency. It can be used for the determination of loureirin A and loureirin B in dragon's blood.