Microfluidic chip immunoassay based on rolling circle amplification and G-quadruplex/Thioflavin T for multiplex detection of CTX I.

A label-free immunoassay based on rolling circle amplification (RCA) and G-quadruplex/Thioflavin T (G4/ThT) is proposed to realize the sensitive detection of carboxy-terminal cross-linked fragment of type I collagen (CTX I) for bone loss. Under the optimal conditions, as low as 38.02 pg/mL of CTX I can be detected. To improve the detecting throughput and simplify the operation, a microfluidic chip was designed, fabricated, and used for CTX I detection based on the proposed assay. The detection can be completed with only a single on-chip magnetic separation step, which was easy to operate, less time-consuming, and has only low reagent consumption. The limit of detection was 131.83 pg/mL by observing with fluorescence microscope. With further improvement of detection equipment, the sensitivity of on-chip detection can be improved. It can be expected that the proposed RCA/G4/ThT immunoassay for sensitive and high-throughput automated detection of CTX I might be chosen as a potential analytical tool for clinical osteoporosis diagnosis and in-orbit bone loss detection.

Oral health self-management barriers among rural older adults in Guangxi, China: A qualitative study.

BACKGROUND: Objective: To understand the barriers associated with self-management of oral health among rural older adults in Guangxi, and to explore the high incidence of oral problems. This information will assist in the formulation of relevant strategies to solve the oral health problems in this population. METHODS: Taking a phenomenological approach, the current status of, and barriers to, oral health self-management in rural older adults from different regions of Guangxi were explored. Participants were purposively selected and interviewed face-to-face. RESULTS: The interviews yielded four overarching themes and six corresponding sub-themes pertaining to barriers in oral health self-management. These included: (1) Older adults' understanding of oral health and disease, perceptions of oral health and their oral health behaviours; (2) Problems in accessing oral health information; (3) Role of family support; and (4) Barriers to healthcare that included access to dental services, oral treatment experience and financial burden of access to dental care. CONCLUSION: Rural older adults in Guangxi face oral health self-management barriers. Improving access to oral healthcare services and changing existing oral health perceptions and habits may assist them in overcoming self-management challenges.

Rapid automatic nucleic acid purification system based on gas-liquid immiscible phase.

The continuous expansion of nucleic acid detection applications has resulted in constant developments in rapid, low-consumption, and highly automated nucleic acid extraction methods. Nucleic acid extraction using magnetic beads across an immiscible phase interface offers significant simplification and parallelization potential. The gas-liquid immiscible phase valve eliminates the requirement for complicated cassettes and is suitable for automation applications. By analyzing the process of magnetic beads crossing the gas-liquid interface, we utilized a low magnetic field strength to drive large magnetic bead packages to cross the gas-liquid interface, providing a solution of high magnetic bead recovery rate for solid-phase extraction with a low-surfactant system based on gas-liquid immiscible phase valve. The recovery rate of magnetic beads was further improved to 90%-95% and the carryover of the reagents was below 1%. Consequently, a chip and an automatic system were developed to verify the applicability of this method for nucleic acid extraction. The Hepatitis B virus serum standard was used for the extraction test. The extraction of four samples was performed within 7 minutes, with nucleic acid recovery maintained above 80% and good purity. Thus, through analysis and experiments, a fast, highly automated, and low-consumption nucleic acid recovery method was proposed in this study.

Nanocellulose: a promising green treasure from food wastes to available food materials.

The synthesis of novel functional materials from abundant food waste resources has great application potentials and ecological benefits. Nanocellulose is a renewable and sustainable polymer that possesses a modifiable surface, excellent mechanical strength, and high aspect ratio, and it is nontoxic. These unique properties garner nanocellulose a promising prospect for multi-various applications including the food industry. This review presents the structural characteristics and advances in the extraction approaches of nanocellulose, with an emphasis in recent progress on the various applications of nanocellulose in the field of food industry. Finally, the environmental and human health issues related to the production of nanocellulose are evaluated. The scheme to extract and produce nanocellulose from food wastes provides a platform for the sustainable utilization of waste biomass. These nanocelluloses exhibit excellent performances in green food packaging materials, emulsion stabilizers, dietary fiber, nutrition delivery and food three-dimensional (3 D) printing hydrogels. To ensure the security and regulatory issues, validated standards to characterize the structure and evaluate its toxicity are still indispensable to achieve the commercialization of nanocellulose in the food industry.

Facile extraction and characterization of cellulose nanocrystals from agricultural waste sugarcane straw.

BACKGROUND: Sugarcane straw is an available but largely ignored lignocellulosic biomass to obtain cellulose nanocrystals (CNCs) with highly crystalline, tunable surface chemistries and a wide-ranging adaptability. Herein, we utilized sugarcane straw to obtain pure cellulose via purification processes, followed by subsequent preparation of CNCs via sulfuric acid hydrolysis. The properties of the purified fibers and obtained CNCs were assessed by their composition, morphology, chemical structure, crystallinity and thermal stability. RESULTS: After the purification process, alkali-treated fibers (ATFs) contained 886.33 ± 1.25 g kg-1 cellulose, and its morphological analysis revealed a smooth and slender fibrous structure. The CNCs obtained by treatment with 64 wt% sulfuric acid at 45 °C for 60 min were isolated in a yield of 21.8%, with a diameter and length of 6 to 10 nm and 160 to 200 nm, respectively. Moreover, crystallinity index of these CNCs reached 62.66%, and thermal stability underwent a two-step degradation. Short-term ultrasonication after hydrolysis was employed to enhance isolation of the CNC particles and improve the anionic charge with higher value -38.00 mV. CONCLUSION: Overall, isolation and characterization results indicated the potential for CNCs preparation using sugarcane straw, in addition to offering a fundamental understanding of this material and indicating potential applications. © 2021 Society of Chemical Industry.

Integrated High-Throughput Centrifugal Microfluidic Chip Device for Pathogen Detection On-Site.

An integrated and high-throughput device for pathogen detection is crucial in point-of-care testing (POCT), especially for early diagnosis of infectious diseases and preventing the spread of infection. We developed an on-site testing platform that utilizes a centrifugal microfluidic chip and automated device to achieve high-throughput detection. The low-power (<32 W), portable (220 mm × 220 mm × 170 mm, 4 kg) device can complete bacterial lysis, nucleic acid extraction and purification, loop-mediated isothermal amplification (LAMP) reaction, and real-time fluorescence detection. Magnetic beads for nucleic acid adsorption can be mixed by applying electromagnetic fields and centrifugal forces, and the efficiency of nucleic acid extraction is improved by 60% compared to the no-mixing group. The automated nucleic acid extraction process achieves equivalent nucleic acid extraction efficiency in only 40% of the time consumed using the kit protocol. By designing the valve system and disc layout, the maximum speed required for the centrifugal microfluidic chip is reduced to 1500 rpm, greatly reducing the equipment power consumption and size. In detecting E. coli, our platform achieves a limit of detection (LOD) of 102 CFU/mL in 60 min. In summary, our active centrifugal microfluidic platform provides a solution for the integration of complex biological assays on turntables, with great potential in the application of point-of-care diagnosis.

Applications of physical and chemical treatments in plant-based gels for food 3D printing.

Extrusion-based three-dimensional (3D) printing has been extensively studied in the food manufacturing industry. This technology places particular emphasis on the rheological properties of the printing ink. Gel system is the most suitable ink system and benefits from the composition of plant raw materials and gel properties of multiple components; green, healthy aspects of the advantages of the development of plant-based gel system has achieved a great deal of attention. However, the relevant treatment technologies are still only at the laboratory stage. With a view toward encouraging further optimization of ink printing performance and advances in this field, in this review, we present a comprehensive overview of the application of diverse plant-based gel systems in 3D food printing and emphasize the utilization of different treatment methods to enhance the printability of these gel systems. The treatment technologies described in this review are categorized into three distinct groups, physical, chemical, and physicochemical synergistic treatments. We comprehensively assess the specific application of these technologies in various plant-based gel 3D printing systems and present valuable insights regarding the challenges and opportunities for further advances in this field.

SIRT2 as a Potential Biomarker in Lung Adenocarcinoma: Implications for Immune Infiltration.

SIRT2 play important roles in cell cycle and cellular metabolism in the development of non-small cell lung cancer (NSCLC), and SIRT2 exhibits its therapeutic effect on NSCLC tumors with high expression of SIRT2. Nevertheless, the clinical relevance of SIRT2 in lung adenocarcinoma (LUAD), particularly its impact on tumor growth and prognostic implications, remains obscure. This investigation entailed a comprehensive analysis of SIRT2 mRNA and protein expression levels in diverse tumor and corresponding healthy tissues, utilizing databases such as TIMER 2.0, UALCAN, and HPA. Prognostic correlations of SIRT2 expression in LUAD patients, stratified by distinct clinicopathological characteristics, were evaluated using the KM Plotter database. Additionally, the TCGA and TIMER 2.0 databases were employed to assess the relationship between SIRT2 and immune infiltration, as well as to calculate immunity, stromal, and estimation scores, thus elucidating the role of SIRT2 in modulating tumor immunotherapy responses. Furthermore, Gene Set Enrichment Analysis (GSEA) was utilized to elucidate SIRT2's biological functions in pan-cancer cells. Our findings revealed a marked reduction in both mRNA and protein levels of SIRT2 in LUAD tumors relative to healthy tissue. Survival analysis indicated that diminished SIRT2 expression correlates with adverse prognostic outcomes in LUAD. Furthermore, SIRT2 expression demonstrated a significant association with various clinicopathologic attributes of LUAD patients, influencing survival outcomes across different clinicopathologic states. Functional enrichment analyses highlighted SIRT2's involvement in cell cycle regulation and immune response. Notably, SIRT2 exhibited a positive correlation with immune cell infiltration, including natural killer (NK) cells, macrophages, and dendritic cells (DCs). In summary, SIRT2 was a potential prognostic biomarker for LUAD and and a new immunotherapy target.

Discovery of novel 3-(1H-pyrazol-4-yl)-1H-indazole derivatives as potent type II TRK inhibitors against acquired resistance.

Tropomyosin receptor kinase (TRK) is a promising target for treating NTRK fusion cancers. The solvent front and xDFG mutations induced by larotrectinib and entrectinib result in acquired resistance in advanced-stage patients. In this study, we report a highly potent and selective type II TRK inhibitor, 40l, developed using a structure-based design strategy. Compound 40l significantly suppressed Km-12, Ba/F3-TRKAG595R, and Ba/F3-TRKAG667C cell proliferation. In biochemical and cellular assays, 40l showed better inhibitory activity against TRKAG667C than that by the positive control, selitrectinib. Additionally, it induced apoptosis of Ba/F3-TRKAG595R and Ba/F3-TRKAG667C cells in a dose-dependent manner. Furthermore, 40l showed good selectivity for a panel of 41 kinases. In vitro assays indicated that 40l possessed outstanding plasma stability and moderate liver microsomal stability. Based on the above results, compound 40l could be further optimized to overcome the solvent front and xDFG TRK mutations.

Enhanced recovery after surgery for percutaneous CT-guided microwave ablation of lung tumors: A single-center retrospective cohort study.

BACKGROUND: The feasibility and safety of enhanced recovery after surgery (ERAS) for percutaneous computed tomography (CT)-guided microwave ablation (MWA) for treating lung nodules remain unclear. METHODS AND MATERIALS: A total of 409 patients with lung tumors treated at the Department of Thoracic Surgery, First Affiliated Hospital of Guangxi Medical University from August 2020 to May 2023 were enrolled. Perioperative data, including baseline characteristics, operation time, postoperative pain score (visual analog scale [VAS]), hospitalization expenses, postoperative complications, total hospital stay, and patient satisfaction, were observed and recorded. RESULTS: No perioperative mortality occurred in either group and complete ablation was achieved in all patients. Patients in the ERAS group had significantly shorter hospital stays (P < 0.001), reduced operation times (P = 0.047), lower hospitalization expenses (P < 0.001), lower VAS scores (P < 0.001), and fewer complications (P = 0.047) compared with the traditional group. CONCLUSIONS: ERAS for percutaneous CT-guided MWA (ERAA) is safe, effective, and feasible for the treatment of lung nodules.

Mild hypothermia pretreatment improves hepatic ischemia-reperfusion injury: A systematic review and meta-analysis of animal experiments.

BACKGROUND AND AIM: Mild hypothermia in hepatic ischemia-reperfusion injury is increasingly being studied. This study aimed to conduct a systematic evaluation of the effectiveness of mild hypothermia in improving hepatic ischemia-reperfusion injury. METHODS: We systematically searched CNKI, WanFang Data, PubMed, Embase, and Web of Science for original studies that used animal experiments to determine how mild hypothermia(32-34°C) pretreatment improves hepatic ischemia-reperfusion injury(in situ 70% liver IR model). The search period ranged from the inception of the databases to May 5, 2023. Two researchers independently filtered the literature, extracted the data, and assessed the risk of bias incorporated into the study. The meta-analysis was performed using RevMan 5.4.1 and Stata 15 software. RESULTS: Eight randomized controlled trials (RCTs) involving a total of 117 rats/mice were included. The results showed that the ALT levels in the mild hypothermia pretreatment group were significantly lower than those in the normothermic control group [Standardized Mean Difference (SMD) = -5.94, 95% CI(-8.09, -3.78), P<0.001], and AST levels in the mild hypothermia pretreatment group were significantly lower than those in the normothermic control group [SMD = -4.45, 95% CI (-6.10, -2.78), P<0.001]. The hepatocyte apoptosis rate in the mild hypothermia pretreatment group was significantly lower than that in the normothermic control group [SMD = -6.86, 95% CI (-10.38, -3.33), P<0.001]. Hepatocyte pathology score in the mild hypothermia pretreatment group was significantly lower than that in the normothermic control group [SMD = -4.36, 95% CI (-5.78, -2.95), P<0.001]. There was no significant difference in MPO levels between the mild hypothermia preconditioning group and the normothermic control group [SMD = -4.83, 95% CI (-11.26, 1.60), P = 0.14]. SOD levels in the mild hypothermia preconditioning group were significantly higher than those in the normothermic control group [SMD = 3.21, 95% CI (1.27, 5.14), P = 0.001]. MDA levels in the mild hypothermia pretreatment group were significantly lower than those in the normothermic control group [SMD = -4.06, 95% CI (-7.06, -1.07) P = 0.008]. CONCLUSION: Mild hypothermia can attenuate hepatic ischemia-reperfusion injury, effectively reduce oxidative stress and inflammatory response, prevent hepatocyte apoptosis, and protect liver function.

Synthesis and characterization of cellulose nanocrystal-Fe composite nanoparticles and their digestion behavior in simulated gastric fluid.

Cellulose nanocrystals (CNC) exhibit great potential as a food emulsifier or functional material template. Herein, CNC-Fe nanoparticles were successfully prepared via an in situ chemical reduction approach. Zeta potential measurements, low-field nuclear magnetic resonance spectroscopy, and atomic force microscopy showed that Fe(III) ions were adsorbed onto CNC when FeCl3 was added to a CNC dispersion. Micromorphological analysis revealed small (diameter = 10.0 ± 2.4 nm) spherical nanoparticles synthesized on the surface of aggregated CNC after the reduction of the Fe(III) ions. Fourier transform infrared spectroscopy revealed an intense peak at 779 cm-1 in the CNC-Fe nanoparticles, which was attributed to FeO stretching vibrations. X-ray photoelectron spectroscopy indicated that the valence state of Fe in CNC-Fe nanoparticles was predominantly ferrous. The synthesized CNC-Fe nanoparticles demonstrated excellent colloidal stability in a dispersion for 21 d and complete, rapid, and spontaneous dissolution in vitro simulated gastric fluid. Our results highlight the potential use of CNC as a template for loading Fe into nanoparticles for Fe fortification in food.

Tailoring the optical and mechanical properties of cellulose nanocrystal film by sugar alcohols and its pH/humidity-responsive behavior.

Cellulose nanocrystals (CNC) have gained widespread attention in intelligent food packaging because of their iridescent optical properties. Here, we report a CNC composite film employing CNC, sugar alcohols (e.g., maltol, erythritol, mannitol, sorbitol, and xylitol) and natural pigment anthocyanins, which has a special iridescent color that can be used as a pH and humidity sensor. The effects of five sugar alcohols with different addition ratios on the structural, optical, and mechanical properties of the CNC films were investigated. The results demonstrated that the addition of sugar alcohol made composite films exhibiting a red-shift of λmax, a more uniform color in visual observation, and a larger pitch. Among them, the CNC-mannitol composite film with a ratio of 10:1 exhibited the best mechanical properties, possessing a tensile stress strength of 57 MPa and toughness of 137 J/m3. Subsequently, anthocyanins were incorporated to this composite film, which showed a marked color change along with the pH from 2 to 12 and exhibited a reversible color change from red to transparent upon a relative humidity change from 35 % to 85 %. Overall, such multi-environment-responsive iridescent films with excellent mechanical properties have a great potential for use in intelligent food packaging applications.

Oscillatory-Flow PCR Microfluidic Chip Driven by Low Speed Biaxial Centrifugation.

PCR is indispensable in basic science and biotechnology for in-orbit life science research. However, manpower and resources are limited in space. To address the constraints of in-orbit PCR, we proposed an oscillatory-flow PCR technique based on biaxial centrifugation. Oscillatory-flow PCR remarkably reduces the power requirements of the PCR process and has a relatively high ramp rate. A microfluidic chip that could perform dispensing, volume correction, and oscillatory-flow PCR of four samples simultaneously using biaxial centrifugation was designed. An automatic biaxial centrifugation device was designed and assembled to validate the biaxial centrifugation oscillatory-flow PCR. Simulation analysis and experimental tests indicated that the device could perform fully automated PCR amplification of four samples in one hour, with a ramp rate of 4.4 ∘C/s and average power consumption of less than 30 W. The PCR results were consistent with those obtained using conventional PCR equipment. Air bubbles generated during amplification were removed by oscillation. The chip and device realized a low-power, miniaturized, and fast PCR method under microgravity conditions, indicating good space application prospects and potential for higher throughput and extension to qPCR.

Design, synthesis and biological evaluation of novel indolin-2-one derivatives as potent second-generation TRKs inhibitors.

Tropomyosin receptor kinases (TRKs) are effective targets for anti-cancer drug discovery. The first-generation type I TRKs inhibitors, larotrectinib and entrectinib, exhibit durable disease control in the clinic. The emergence of acquired resistance mediated by secondary mutations in the TRKs domain significantly reduces the therapeutic efficacy of these two drugs, indicating an unmet clinical need. In this study, we designed a potent and orally bioavailable TRK inhibitor, compound 24b, using a molecular hybridization strategy. Compound 24b exhibited significant inhibitory potency against multiple TRK mutants in both biochemical and cellular assays. Furthermore, compound 24b induced apoptosis of Ba/F3-TRKAG595R and Ba/F3-TRKAG667C cells in a dose-dependent manner. Additionally, compound 24b exhibited moderate kinase selectivity. In vitro stability revealed that compound 24b showed excellent plasma stability (t1/2 > 289.1 min) and moderate liver microsomal stability (t1/2 = 44.3 min). Pharmacokinetic studies have revealed that compound 24b is an orally bioavailable TRK inhibitor with a good oral bioavailability of 116.07%. These results indicate that compound 24b be used as a lead molecule for further modifications to overcome drug-resistant mutants of TRK.

Prediction of lymph node metastasis of lung squamous cell carcinoma by machine learning algorithm classifiers.

BACKGROUND: Lymph node metastasis (LNM) is an essential factor affecting the prognosis of patients with lung squamous cell carcinoma (LUSC), as well as a critical consideration for the choice of treatment strategy. Exploring effective methods for predicting LNM in LUSC may benefit clinical decision making. MATERIALS AND METHODS: We used data collected from the Surveillance, Epidemiology, and End Results (SEER) database to develop machine learning algorithm classifiers, including boosted trees (BTs), based on the primary clinical parameters of patients to predict LNM in LUSC. Training on a large-sample training cohort (n = 8,063) allowed for the construction of several concise classifiers for LNM prediction in LUSC, which were then validated using test and in-house cohorts (n = 2,017 and 57, respectively). RESULTS: The six classifiers established in this research enabled distinction between patients with and without LNM. Among these classifiers, the BT classifier was the top performer, with accuracy, F1 scores, precision, recall, sensitivity, and specificity values of 0.654, 0.621, 0.654, 0.592, 0.592, and 0.711, respectively. The precision recall (PR) and receiver operating characteristic (ROC) (with area under the curve = 0.714) curves also supported this result, which was validated by the in-house cohort. Notably, the tumor stage was a critical factor in determining LNM in patients with LUSC. CONCLUSIONS: The use of classifiers, especially the BT classifier, may serve as a useful tool for improving clinical precision and individualized treatment of patients with LUSC.

Studies into interactions and interfacial characteristics between cellulose nanocrystals and bovine serum albumin.

This study investigates the interactions between cellulose nanocrystals (CNCs) and bovine serum albumin (BSA) under different pH conditions. A multiscale technique was employed to characterize the CNCs and BSA at pH 7 and pH 3. ζ-Potential measurement and UV-vis spectroscopy demonstrated strong interactions between CNCs and BSA at pH 3, whereat they have opposite charges. Interfacial tensiometry showed a deficiency in the surface activity of the CNCs and indicated that BSA dominated the interface behavior in their complex. Quartz crystal microbalance with dissipation revealed that the sequential adsorption of BSA and CNCs produced viscoelastic bilayers at pH 3, and the mass adsorbed was âˆ¼ 28 times that adsorbed at pH 7. Molecular dynamics simulations indicated that the key interactions between the two materials were produced between the hydrophobic CNC surface and the BSA domain IIA region. These results provide interesting insights into the design of complex food emulsions and fluid interfaces.

Effects of lipid type and toxicological properties on the digestion of cellulose nanocrystals in simulated gastrointestinal tract.

This study aimed to understand the impact of different oil types on the cellulose nanocrystals (CNCs) to modulate lipid digestion and the in vitro gastrointestinal toxicity of CNCs in food systems. We explored the ability of CNCs to modulate lipid digestion in a simulated gastrointestinal system and monitored the gastrointestinal fate of CNC-based emulsions with different oil types. Finally, a small intestine epithelial model was used to evaluate the influence of cytotoxicity. The results suggested that the addition of 0.6 wt% CNCs in the high-fat food model reduced the hydrolysis of free fatty acids (FFAs) from triglycerides by 37.8% after the small intestine phase. CNCs showed the best effect in reducing lipid digestion in emulsions with high unsaturation triglycerides. In addition, the toxicology results suggest that 0.6 wt% CNCs had only a slight effect on reactive oxygen species and cytotoxicity, and no significant change in cell-layer integrity.

Thromboelastography for the Prevention of Perioperative Venous Thromboembolism in Orthopedics.

We have reviewed a large number of relevant literature to determine the deficiencies of orthopedics in the diagnosis and prevention of venous thromboembolism（VTE）events during the perioperative period, and found that the TEG technology has been widely used after liver transplantation, which may make up for the deficiencies. This review expounds the detection principle and latest thromboelastography (TEG) development, and highlights the advantages of TEG over previous screening methods in diagnosing hypercoagulability. By analyzing the correlation and consistency between TEG and conventional coagulation test, reliable indexes for diagnosing hypercoagulability and important parameters for guiding perioperative anticoagulation treatment were summarized. Furthermore, our work contributes to further studies of TEG in orthopedics. Based on the research results, we believe that TEG may help orthopedists to identify and predict VTE events, use anticoagulants, eventually reduce the occurrence of VTE events.

Directed evolution of the genetically encoded zinc(II) FRET sensor ZapCY1.

Zinc(II) ions (Zn2+) play an essential role in living systems, with their delicate concentration balance differing among the various intracellular organelles. The spatiotemporal distribution and homeostasis of Zn2+ can be monitored through photoluminescence imaging using zinc sensors. Among such biosensors, genetically encoded fluorescent sensor proteins are attractive tools owing to their subcellular localization advantage and high biocompatibility. However, the limited fluorescent properties of these proteins, such as their insufficient quantum yield and dynamic range, restrict their practical use. In this study, we developed an expression-screening-directed evolution system and used it to improve ZapCY1, a genetically encoded fluorescence resonance energy transfer (FRET) sensor. After four rounds of directed evolution, the FRET dynamic range of the modified sensor (designated ZapTV-EH) was increased by 1.5-1.7-fold. With its enhanced signal-to-noise ratio and ability to detect a wide Zn2+ concentration range, ZapTV-EH proves to be a better visualization tool for monitoring Zn2+ at the subcellular level. Combined with the simplified subcloning and expression steps and sufficient mutant libraries, this directed evolution system may provide a more simple and efficient way to develop and optimize genetically encoded FRET sensors through high-throughput screening.