Investigation of the synergistic effect mechanism underlying sequential use of palbociclib and cisplatin through integral proteomic and glycoproteomic analysis.

Chemoresistance largely hampers the clinical use of chemodrugs for cancer patients, combination or sequential drug treatment regimens have been designed to minimize chemotoxicity and resensitize chemoresistance. In this work, the cytotoxic effect of cisplatin was found to be enhanced by palbociclib pretreatment in HeLa cells. With the integration of liquid chromatography-mass spectrometry-based proteomic and N-glycoproteomic workflow, we found that palbociclib alone mainly enhanced the N-glycosylation alterations in HeLa cells, while cisplatin majorly increased the different expression proteins related to apoptosis pathways. As a result, the sequential use of two drugs induced a higher expression level of apoptosis proteins BAX and BAK. Those altered N-glycoproteins induced by palbociclib were implicated in pathways that were closely associated with cell membrane modification and drug sensitivity. Specifically, the top four frequently glycosylated proteins FOLR1, L1CAM, CD63, and LAMP1 were all associated with drug resistance or drug sensitivity. It is suspected that palbociclib-induced N-glycosylation on the membrane protein allowed the HeLa cell to become more vulnerable to cisplatin treatment. Our study provides new insights into the mechanisms underlying the sequential use of target drugs and chemotherapy drugs, meanwhile suggesting a high-efficiency approach that involves proteomic and N-glycoproteomic to facilitate drug discovery.

A deep learning-based algorithm for pulmonary tuberculosis detection in chest radiography.

In tuberculosis (TB), chest radiography (CXR) patterns are highly variable, mimicking pneumonia and many other diseases. This study aims to evaluate the efficacy of Google teachable machine, a deep neural network-based image classification tool, to develop algorithm for predicting TB probability of CXRs. The training dataset included 348 TB CXRs and 3806 normal CXRs for training TB detection. We also collected 1150 abnormal CXRs and 627 normal CXRs for training abnormality detection. For external validation, we collected 250 CXRs from our hospital. We also compared the accuracy of the algorithm to five pulmonologists and radiological reports. In external validation, the AI algorithm showed areas under the curve (AUC) of 0.951 and 0.975 in validation dataset 1 and 2. The accuracy of the pulmonologists on validation dataset 2 showed AUC range of 0.936-0.995. When abnormal CXRs other than TB were added, AUC decreased in both human readers (0.843-0.888) and AI algorithm (0.828). When combine human readers with AI algorithm, the AUC further increased to 0.862-0.885. The TB CXR AI algorithm developed by using Google teachable machine in this study is effective, with the accuracy close to experienced clinical physicians, and may be helpful for detecting tuberculosis by CXR.

A retrospective study of 3D laparoscopy and 2D laparoscopy in transabdominal preperitoneal (TAPP) for unilateral inguinal hernia in elderly patients.

This study aims to explore the safety and efficacy of 3D laparoscopy in elderly patients undergoing transabdominal preperitoneal (TAPP) surgery for inguinal hernia. Patients were divided into two groups based on the laparoscopic equipment used during surgery. Clinical data preoperatively, intraoperatively, and postoperatively were collected and subjected to statistical analysis. From January 2020 to August 2023, a total of 127 patients with primary unilateral inguinal hernia were evaluated in this study, 61 in the 3D TAPP group and 66 in the 2D TAPP group. There were no statistically significant differences in baseline data, including average age, gender distribution, BMI index, hernia type, hernia defect size and location, comorbidities, and usage of anticoagulant drugs between the two groups (P > 0.05). In terms of operative indicators, the 3D group showed shorter mean operation time (51.61 ± 7.16 min vs. 78.59 ± 13.51 min, P < 0.001), mean mesh placement time (6.07 ± 1.40 min vs. 9.77 ± 1.21 min, P < 0.001), and mean peritoneal suture time (7.34 ± 1.85 min vs. 9.73 ± 1.32 min, P < 0.001) compared to the 2D group. However, there were no statistically significant differences in mean blood loss, postoperative pain scores, postoperative hospital stay, and total hospital costs between the two groups (P > 0.05). The incidence of postoperative complications did not differ significantly between the two groups (P > 0.05). No adverse reactions such as dizziness or nausea were reported by surgeons during the procedures in either group. Three-dimensional laparoscopy in TAPP surgery provides high-definition, three-dimensional surgical images, reducing the difficulty of operations and effectively shortening the operation time.

Numerical Analysis and Quantification of Transfer Efficiency Coupled with Capillary and Quadrupole Ion Guide in an API-MS System.

Ion trajectory simulation is a significant and useful tool for understanding ion transfer mechanisms within the first vacuum region of the atmospheric pressure ionization mass spectrometer (API-MS). However, the complex dynamic gas field and wide pressure range lead to inaccurate simulation and huge computational costs. In this work, a novel electrohydrodynamic simulation called the statistical diffusion-hard-sphere (SDHS) mixed collision model was developed for characterizing the ion trajectories. For the first time, the influence of the dynamic pressure on the ion trajectory is considered for simulation, which helps to avoid an intolerable computational cost. Comparing with the conventional Monte Carlo collision model, the SDHS method helps to improve the calculation accuracy of ion trajectories under the first vacuum region and reduce the computational cost for at least 12-folds. Simulation results showed that the maximum ion loss came from the gap of the electrodes. The distance of the capillary-quadrupole ion guide was also a non-negligible factor. The trend of quantitative experimental results matches the SDHS simulation results. The maximum ion transfer efficiencies of quantitative experiment and simulation were 55% and 52%, respectively. Moreover, three ions, caffeine, reserpine, and Ultramark 1621, were measured for evaluating the applicability of SDHS in real API-MS. The trend of experimental results showed good agreement with that of computation. And the results of caffeine further illustrated the reason that the small mass ion transfer efficiency decreased with increasing radio frequency voltage. SDHS method is expected to be useful in the design of ion guides for further improvement of the sensitivity of API-MS.

Stimuli-Responsive Ion Transport Regulation in Nanochannels by Adhesion-Induced Functionalization of Macroscopic Outer Surface.

Responsive regulation of ion transport through nanochannels is crucial in the design of smart nanofluidic devices for sequencing, sensing, and water-energy nexus. Functionalization of the inner wall of the nanochannel enhances interaction with ions and fluid but restricts versatile chemical approaches and accurate characterizations of fluidic interfaces. Herein, we reveal a responsive regulating mechanism of ion transport through nanochannels by polydopamine (PDA)-induced functionalization on the macroscopic outer surface of nanochannels. Responsive molecules were codeposited with PDA on the outer surface of nanochannels and formed a valve of nanometer thickness to manually manipulate ion transport by changing its gap spacing, surface charge, and wettability under external stimulus. The response ratio can be up to 100-fold by maximizing the proportion of responsive molecules on the outer surface. Laminating the codepositions of different responsive molecules with PDA on the channel's outer surface produces multiple responses. A nearly universal adhesion of PDA with responsive molecules on the open outer surface induces nanochannels responsive to different external stimuli with variable response ratios and arbitrary combinations. The results challenge the primary role of functionalization on the nanoconfined interface of nanofluidics and open opportunities for developing new-style nanofluidic devices through the functionalization of macroscopic interface.

Direct Oxidative Cyclization of 3-Arylpropionic Acids to 3,4-Dihydrocoumarins: Reinvestigation of the Reaction Mechanism.

Instigated by olfactory analysis of odorant molecules, the constitutions of 3,4-dihydrocoumarins prepared by PIFA-based oxidative cyclizations of 3-arylpropionic acids were revised by means of 2D NMR and X-ray analysis. Supported by computational analysis, the migratory mechanism of intermediate spirolactonic cations has been amended: 1,2-alkyl shifts instead of 1,2-carboxylic shifts were selectively obtained.

Integrated plasma proteomics and N-glycoproteomics reveals alterations in the N-glycosylation in Chinese hepatocellular carcinoma patients.

Hepatocellular carcinoma (HCC) is a life-threatening disease that presents diagnostic challenges due to the absence of reliable biomarkers. Recently, plasma proteomics and glycoproteomics have emerged as powerful tools for identifying potential diagnostic biomarkers for various diseases. In this study, we conducted a comprehensive proteomic and glycoproteomic analysis of plasma samples from 11 HCC patients and 11 healthy control (HC) individuals. We identified 20 differentially expressed (DE) proteins and 32 DE intact glycosylated peptides (IGPs) that can effectively differentiate between HCC patients and HC samples. Our findings demonstrate that IGP profiles had better predictive power than protein profiles for screening HCC. Pathways associated with DE proteins and IGPs were identified. It was reported that the protein expression level of galectin 3 binding protein (LGALS3BP) and its N-linked glycosylation at the N398 and N551 sites might serve as valuable candidates for HCC diagnosis. These results highlight the importance of N-glycoproteomics in advancing our understanding of HCC and suggest possible candidates for the future diagnosis of this disease.

Nakazawaea tricholomae f.a., sp. nov., a Novel Ascomycetous Yeast Species Isolated from Two Mushroom Species in China.

Two yeast strains designated as 20-27-1 and 20-28 were isolated from the fruiting bodies of Tricholoma gambosum and Marasmius maximus, respectively, which were collected in Wudaogou, Weichang county, Chengde area, Hebei Province, China. The multi-locus analysis of the sequences of the rDNA ITS, D1/D2 LSU, and SSU regions, together with partial sequences of two protein-coding genes RPB1 and TEF1 indicates that the two strains are closely related to Nakazawaea ernobii and Nakazawaea holstii, showing the similarity values of 99.3-98.7%, 97.2-97.1%, 91.9-92.5%, and 84.6% in D1/D2 LSU, ITS, TEF1, and RPB1, respectively. Physiologically, the two strains are different from N. ernobii and N. holstii in the assimilation of melibiose, inulin, and DL-lactic acid. Both the phenotypic and phylogenetic analyses indicate that those two strains represent a novel species in the genus Nakazawaea, for which the name Nakazawaea tricholomae f.a., sp. nov. (Fungal Names: FN 571492) is proposed.

CD206+ M2-like macrophages protect against intervertebral disc degeneration partially by targeting R-spondin-2.

OBJECTIVE: This study aimed to explore the specific function of M2 macrophages in intervertebral disc degeneration (IDD). METHODS: Intervertebral disc (IVD) samples from normal (n = 4) and IDD (n = 6) patients were collected, and the expression of M2-polarized macrophage marker, CD206, was investigated using immunohistochemical staining. Nucleus pulposus cells (NPCs) in a TNF-α environment were obtained, and a mouse caudal IVD puncture model was established. Mice with Rheb deletions, specifically in the myeloid lineage, were generated and subjected to surgery-induced IDD. IDD-induced damage and cell apoptosis were measured using histological scoring, X-ray imaging, immunohistochemical staining, and TdT-mediated dUTP nick end labeling (TUNEL) assay. Finally, mice and NPCs were treated with R-spondin-2 (Rspo2) or anti-Rspo2 to investigate the role of Rspo2 in IDD. RESULTS: Accumulation of CD206 in human and mouse IDD tissues was detected. Rheb deletion in the myeloid lineage (RheBcKO) increased the number of CD206+ M2-like macrophages (mean difference 18.6% [15.7-21.6%], P < 0.001), decreased cell apoptosis (mean difference -15.6% [-8.9 to 22.2%], P = 0.001) and attenuated the IDD process in the mouse IDD model. NPCs treated with Rspo2 displayed increased extracellular matrix catabolism and apoptosis; co-culture with a conditioned medium derived from RheBcKO mice inhibited these changes. Anti-Rspo2 treatment in the mouse caudal IVD puncture model exerted protective effects against IDD. CONCLUSIONS: Promoting CD206+ M2-like macrophages could reduce Rspo2 secretion, thereby alleviating experimental IDD. Rheb deletion may help M2-polarized macrophages accumulate and attenuate experimental IDD partially by inhibiting Rspo2 production. Hence, M2-polarized macrophages and Rspo2 may serve as therapeutic targets for IDD.

Halogenated aliphatic and phenolic disinfection byproducts in chlorinated and chloraminated dairy wastewater: Occurrence and ecological risk evaluation.

The increasing demand for dairy products has led to the production of a large amount of wastewater in dairy plants, and disinfection is an essential treatment process before wastewater discharge. Disinfection byproducts (DBPs) in disinfected dairy wastewater may negatively influence the aquatic organisms in receiving water. During chlorine and chloramine disinfection of dairy wastewater, the concentrations of aliphatic DBPs increased from below the detection limits to 485.1 µg/L and 26.6 µg/L, respectively. Brominated and iodinated phenolic DBPs produced during chlor(am)ination could further react with chlorine/chloramine to be transformed. High level of bromide in dairy wastewater (12.9 mg/L) could be oxidized to active bromine species by chlorine/chloramine, promoting the formation of highly toxic brominated DBPs (Br-DBPs), and they accounted for 80.3% and 71.1% of the total content of DBPs in chlorinated and chloraminated dairy wastewater, respectively. Moreover, Br-DBPs contributed 49.9-75.9% and 34.2-96.4% to the cumulative risk quotient of DBPs in chlorinated and chloraminated wastewater, respectively. The cumulative risk quotient of DBPs on green algae, daphnid, and fish in chlorinated wastewater was 2.8-11.4 times higher than that in chloraminated wastewater. Shortening disinfection time or adopting chloramine disinfection can reduce the ecological risks of DBPs.

Development of cancer biomarker heat shock protein 90α certified reference material using two different isotope dilution mass spectrometry techniques.

Heat shock protein 90α (HSP90α) has been regarded as an important indicator for judging tumor metastasis and prognosis due to its significant upregulation in various tumors. Therefore, the accurate quantification of HSP90α is of great significance for clinical diagnosis and therapy of cancers. However, the lack of HSP90α certified reference material (CRM) leads to the accuracy and consistency of quantification methods not being effectively evaluated. Besides, quantitative results without traceability make comparisons between different studies difficult. In this study, an HSP90α solution CRM was developed from the recombinant protein raw material. The recombinant protein is a dimer, and the purity of the CRM candidate reached 96.71%. Both amino acid analysis-isotope dilution mass spectrometry (AAA-IDMS) and unique peptide analysis-isotope dilution mass spectrometry (UPA-IDMS) were performed to measure the content of HSP90α in the solution CRM candidate, and the certified value was assessed to be 66.2 ± 8.8 µg/g. Good homogeneity of the CRM was identified, and the stability examination suggested that the CRM was stable for at least 4 months at - 80 °C and for 7 days at 4 °C. With traceability to SI unit (kg), this CRM has potential to help establish a metrological traceability chain for quantification of HSP90α, which will make the quantification results standardized and comparable regardless of the quantitative methods.

Comparative analysis of laparoscopic choledocholithiasis and ERCP treatment after cholecystectomy.

OBJECTIVE: To compare the overall efficacy of laparoscopic common bile duct exploration(LCBDE) with endoscopic retrograde cholangiopancreatography (ERCP ) after cholecystectomy. METHODS: From January 2017 to July 2021, Seventy patients with Choledocholithiasis after cholecystectomy who were admitted to our hospital were selected and divided into ERCP and LCBDE groups. comparison of baseline characteristics, clinical efficacy and postoperative complications between the ERCP and LCBDE. RESULTS: ①The overall efficacy rate of LCBDE was 97.1%, while the overall efficacy rate in the ERCP group was 76.6%. The LCBDE group demonstrated a significantly higher overall effective rate compared to the ERCP group, with a statistically significant difference (p < 0.05). ②The preoperative and postoperative complications of the LCBDE group were visibly lower than the other group (P < 0.05). The postoperative time to oral intake, postoperative ventilation time, length of hospital stay, and hospital costs were higher in the ERCP group compared to the LCBDE group, with a statistically significant difference (P < 0.05). CONCLUSION: In the treatment of common bile duct stones after cholecystectomy, LCBDE is a superior choice compared to ERCP in terms of stone diameter, quantity, clearance rate, and hospital costs.

Precise Control of Trypsin Immobilization by a Programmable DNA Tetrahedron Designed for Ultrafast Proteome Digestion and Accurate Protein Quantification.

In proteomics research, with advantages including short digestion times and reusable applications, immobilized enzyme reactors (IMERs) have been paid increasing attention. However, traditional IMERs ignore the reasonable spatial arrangement of trypsin on the supporting matrixes, resulting in the partial overlapping of the active domain on trypsin and reducing digesting efficiency. In this work, a DNA tetrahedron (DNA TET)-based IMER Fe3O4-GO-AuNPs-DNA TET-Trypsin was designed and prepared. The distance between vertices of DNA TETs effectively controls the distribution of trypsin on the nanomaterials; thus, highly efficient protein digestion and accurate quantitative results can be achieved. Compared to the in-solution digestion (12-16 h), the sequence coverage of bovine serum albumin was up to 91% after a 2-min digestion by the new IMER. In addition, 3328 proteins and 18,488 peptides can be identified from HeLa cell protein extract after a 20-min digestion. For the first time, human growth hormone reference material was rapidly and accurately quantified after a 4-h digestion by IMER. Therefore, this new IMER has great application potential in proteomics research and SI traceable quantification.

Maternal gestational diabetes mellitus and the childhood asthma in offspring: a meta-analysis.

BACKGROUND: Maternal diabetes might be related to a high risk of allergic disease in offspring. However, it remains unknown if maternal gestational diabetes mellitus (GDM) is also associated with a high incidence of childhood asthma in offspring. A systematic review and meta-analysis was performed to investigate the above association. METHODS: Relevant observational studies were obtained by search of electronic databases including Medline, Embase, Cochrane Library, and Web of Science. A randomized-effects model was selected to pool the data by incorporating the influence of potential heterogeneity. The Newcastle-Ottawa Scale was used for study quality evaluation. Subgroup analyses were performed to evaluate the potential influences of study characteristics on the outcome. RESULTS: Ten datasets from seven moderate to high quality cohort studies, involving 523,047 mother-child pairs were included in the meta-analysis. Overall, maternal GDM was associated with a higher risk of childhood asthma in offspring (risk ratio [RR]: 1.22, 95% confidence interval [CI]: 1.07 to 1.39, p = 0.003; I2 = 30%). Subgroup analyses showed that the association was not significantly affected by study design, validation methods for GDM, or diagnostic strategy for asthma (p for subgroup analyses all > 0.05). The association between maternal GDM and asthma in offspring was more remarkable after adjusting maternal body mass index in early pregnancy (RR: 1.50 versus 1.06, p < 0.001), but significantly weakened after adjusting hypertensive disorders during pregnancy (RR: 1.08 versus 1.50, p = 0.001). CONCLUSIONS: Maternal GDM may be associated with an increased incidence of childhood asthma in offspring.

Nanoliter atmospheric pressure photoionization-mass spectrometry for direct bioanalysis of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are a class of low-polarity environmental contaminants that have severe carcinogenic effects and have drawn worldwide attention. However, there remain challenges for current mass spectrometric ionization techniques in the analysis of low-polarity compounds in small-volume biosamples, such as single cells. In this work, we developed a nanoliter atmospheric pressure photoionization (nano-APPI) source and optimized its parameters for the detection of PAHs in small-volume samples. We evaluated the ionization performance of the source in direct and auxiliary gas-assisted photoionization modes and analyzed different PAH compounds as well as spiked biosamples. By combining the advantages of nano-electrospray ionization (nano-ESI) and atmospheric pressure photoionization (APPI), our newly developed nano-APPI source achieved high sensitivity for the analysis of PAHs down to the fmol level. Compared to conventional atmospheric pressure chemical ionization (APCI), the detection limit of PAHs was increased by 1-2 orders of magnitude. By optimizing various parameters, we achieved highly efficient ionization of PAHs, effective analysis of PAHs in mixed components, and sensitive detection of low-abundance PAHs in single-cell samples. Our optimized nano-APPI source was successfully applied for the sensitive analysis of PAHs in complex biological samples. Based on our study, we believe that nano-APPI holds great promise for toxicological studies on complex biological samples. The present work has implications for improving the detection sensitivity of low-polarity environmental contaminants and advancing the field of MS-based analysis of small-volume biosamples.

Observing ion diffusion and reciprocating hopping motion in water.

When an ionic crystal dissolves in solvent, the positive and negative ions associated with solvent molecules release from the crystal. However, the existing form, interaction, and dynamics of ions in real solution are poorly understood because of the substantial experimental challenge. We observed the diffusion and aggregation of polyoxometalate (POM) ions in water by using liquid phase transmission electron microscopy. Real-time observation reveals an unexpected local reciprocating hopping motion of the ions in water, which may be caused by the short-range polymerized bridge of water molecules. We find that ion oligomers, existing as highly active clusters, undergo frequent splitting, aggregation, and rearrangement in dilute solution. The formation and dissociation of ion oligomers indicate a weak counterion-mediated interaction. Furthermore, POM ions with tetrahedral geometry show directional interaction compared with spherical ions, which presents structure-dependent dynamics.

Oxidative Three-Component Selenofunctionalization of Alkenes: Convenient Access to Vicinally Functionalized Selenides.

Three-component selenofunctionalization processes of olefins, diselenides and sulfonamides, water, alcohols, or acids utilizing 1-fluoropyridinium triflate (FP-OTf) as a reaction promoter are reported. Under the optimal conditions, a broad range of vicinally functionalized selenide derivatives was accessible with high yields and excellent functional group compatibilities. Mechanistic studies revealed that the FP-OTf played a key role in this selenofunctionalization process.

Quantification of 25OHD in serum by ID-LC-MS/MS based on oriented immobilization of antibody on magnetic materials.

Magnetic nanomaterials are widely used, but co-adsorption of impurities will lead to saturation. In this study, the aim was to prepare a magnetic nano-immunosorbent material based on orienting immobilization that can purify and separate 25-hydroxyvitamin D (25OHD) from serum and provides a new concept of sample pretreatment technology. Streptococcus protein G (SPG) was modified on the surface of the chitosan magnetic material, and the antibody was oriented immobilized using the ability of SPG to specifically bind to the Fc region of the monoclonal antibody. The antigen-binding domain was fully exposed and made up for the deficiency of the antibody random immobilization. Compared with the antibody in the random binding format, this oriented immobilization strategy can increase the effective activity of the antibody, and the amount of antibody consumed is saved to a quarter of the former. The new method is simple, rapid, and sensitive, without consuming a lot of organic reagents, and can enrich 25OHD after simple protein precipitation. Combining with liquid chromatography-tandem mass spectrometry (LC-MS/MS), the analysis can be completed in less than 30 min. For 25OHD2 and 25OHD3, the LOD was 0.021 and 0.017 ng mL-1, respectively, and the LOQ was 0.070 and 0.058 ng mL-1, respectively. The results indicated that the magnetic nanomaterials based on oriented immobilization can be applied as an effective, sensitive, and attractive adsorbent to the enrichment of serum 25OHD.

Evolution of Mass Spectrometry Instruments and Techniques for Blood Proteomics.

Mass spectrometry (MS)-based blood proteomics is a crucial research focus in identifying disease biomarkers. Blood serum or plasma is the most commonly used sample for such analysis; however, it presents challenges due to the complexity and dynamic range of protein abundance. Despite these difficulties, the development of high-resolution MS instruments has made comprehensive investigation of blood proteomics possible. The evolution of time-of-flight (TOF) or Orbitrap MS instruments has played a significant role in the field of blood proteomics. These instruments are now among the most prominent techniques for blood proteomics due to their sensitivity, selectivity, fast response, and stability. For optimal results, it is necessary to eliminate high-abundance proteins from the blood sample, to maximize the depth coverage of the blood proteomics analysis. This can be achieved through various methods, including commercial kits, chemically synthesized materials, and MS technologies. This paper reviews recent advancements in MS technology and its remarkable applications in biomarker discovery, particularly in the areas of cancer and COVID-19 studies.