Analysis of the effect of cations on protein conformational stability using solid-state nanopores.

The conformation of proteins is closely related to their biological functions, and it is affected by many factors, including the type of cations in solution. However, it is difficult to detect the conformational changes of a protein in situ. As a single-molecule sensing technology, nanopores can convert molecular structural information into analyzable current signals within a reasonable time range. Herein, we detect and analyze the effects of two different types of monovalent cations (Na+ and Li+) on a model protein bovine serum albumin (BSA) conformation using SiNx nanopores with different diameters. The quantitative analysis results show that the excluded volume of BSA in LiCl salt solutions is larger than the value in NaCl solution, indicating that Li+ is more prone to unfolding the proteins and making them unstable. This study demonstrated that nanopores enable the in situ detection of the structure of proteins at the single-molecule level and provide a new approach for the quantitative analysis of proteins.

Optical coherence tomography parameters as prognostic factors for stereopsis after vitrectomy for unilateral epiretinal membrane: a cohort study.

This retrospective cohort study explored the relationship between monocular and interocular optical coherence tomography (OCT) parameters and stereopsis in 56 patients undergoing pars plana vitrectomy (PPV) for unilateral idiopathic epiretinal membrane (IERM). IERM impairs visual functions, with symptoms ranging from asymptomatic to severe impairment. Despite established surgical interventions, including PPV with membrane peeling, the impact on advanced three-dimensional visual functions such as stereopsis remains inadequately investigated. All subjects were assessed for stereopsis, visual acuity, and metamorphopsia, alongside spectral domain OCT parameters. These visual functions significantly improved 3-month postoperatively. Central retinal thickness at the fovea, parafovea, and perifovea (CFT, CRT-3 mm, and CRT-6 mm), ectopic inner foveal layer thickness, and retinal layer thickness notably decreased 1 week to 3 months after surgery. The interocular difference in OCT parameters between bilateral eyes was included as a parameter. Baseline CRT-3 mm difference and inner nuclear layer (INL) thickness were independently correlated with postoperative stereopsis on the Titmus Stereo Test, while baseline CRT-6 mm difference and INL thickness were independently related to stereopsis on the TNO stereotest. This study highlights the substantial enhancement in stereopsis post-IERM surgery, with both interocular and monocular OCT parameters independently influencing postoperative stereopsis. These findings underscore the importance of retinal microstructures in assessing and predicting stereopsis in IERM patients after vitrectomy.

Pseudomonas weihenstephanensis through the iron metabolism pathway promotes in situ spoilage capacity of prepared beef steaks during cold storage.

In this study, we evaluated the histomorphology, reactive oxygen species (ROS), protein degradation, and iron metabolism characteristics and differential expression analysis of genes for siderophores synthesis and protease secretion in prepared beef steaks inoculated alone or co-inoculated with P. weihenstephanensis, B. thermotrichothrix and M. caseolyticus at 4 °C for 12 days. The results showed that the P. weihenstephanensis was the key bacteria that degraded protein in the process of prepared beef steaks spoilage, which led to protein oxidation by promoting ferritin degradation to release free iron and inducing ROS accumulation. The highest expression of FpvA and AprE was detected in the P. weihenstephanensis group by comparing qRT-PCR of the different inoculation groups. Both qRT-PCR and Western blot revealed that ferritin heavy polypeptide and ferritin light chain polypeptide gene and protein expressions were significantly higher in the P. weihenstephanensis inoculation group compared to the other inoculation groups. Results suggested that FpvA and AprE might play roles in meat spoilage and were potential positional, physiological and functional candidate genes for improving the quality traits of prepared beef steaks. This work may provide insights on controlling food quality and safety by intervening in spoilage pathways targeting iron carrier biosynthesis or protease secretion genes.

Carbon availability and microbial activity manipulate the temperature sensitivity of anaerobic degradation in a paddy soil profile.

Soil contains a substantial amount of organic carbon, and its feedback to global warming has garnered widespread attention due to its potential to modulate atmospheric carbon (C) storage. Temperature sensitivity (Q10) has been widely utilized as a measure of the temperature-induced enhancement in soil organic carbon (SOC) decomposition. It is currently rare to incorporate Q10 of CO2 and CH4 into the study of waterlogged soil profiles and explore the possibility of artificially reducing Q10 in rice fields. To investigate the key drivers of Q10, we collected 0-1 m paddy soil profiles, and stratified the soil for submerged anaerobic incubation. The relationship between SOC availability, microbial activity, and the Q10 of CO2 and CH4 emissions was examined. Our findings indicate that as the soil layer deepens, soil C availability and microbial activity declined, and the Q10 of anaerobic degradation increased. Warming increased C availability and microbial activity, accompanied by weakened temperature sensitivity. The Q10 of CO2 correlated strongly with soil resistant C components, while the Q10 of CH4 was significantly influenced by labile substrates. The temperature sensitivity of CH4 (Q10 = 3.99) was higher than CO2 emissions (Q10 = 1.78), indicating the need for greater attention of CH4 in predicting warming's impact on anaerobic degradation in rice fields. Comprehensively assessing CO2 and CH4 emissions, the 20-40 cm subsurface soil is the most temperature-sensitive. Despite being a high-risk area for C loss and CH4 emissions, management of this soil layer in agriculture has the potential to reduce the threat of global warming. This study underscores the importance of subsurface soil in paddy fields, advocating greater attention in scientific simulations and predictions of climate change.

The impact of preoperative parameters on postoperative foveal displacement in idiopathic macular hole.

This study examined the effect of vitrectomy combined with internal limiting membrane (ILM) peeling on foveal displacement in 42 eyes with idiopathic macular hole (IMH). A retrospective analysis was conducted to measure various macular hole parameters before surgery, including basal diameter, minimum diameter, hole height, and areas affected by traction such as macular hole area (MHA), macular hole cystoid space area (MHCSA), macular hole retinal area (MHRA), and total area (TA). The results showed a postoperative shift of the fovea towards the optic disc in all cases. Notably, the extent of foveal displacement was significantly linked to the preoperative basal diameter (rs = 0.405, P = 0.008) but not to other preoperative parameters or postoperative visual acuity. Furthermore, the study found that the temporal side of the macular hole was more affected by traction than the nasal side preoperatively, leading to greater postoperative displacement (All P < 0.05).

Efficient Fresh Lamp Light-Harvesting Films with the Self-Activating Continuous and Recyclable Bactericidal Ability for Ultrapersistent Freshness of Perishable Muscle Food.

A large quantity of perishable muscle food is being wasted due to harmful bacteria infestation during the sales and circulation each year and facing challenges. In this study, a self-activated bactericidal active film (PLA/g-C3N4@PCN-224) responsive to fresh lamp light was prepared, which showed excellent hydrophobicity, water vapor resistance, and thermal stability. Due to the synergistic effect between light-induced reactive oxygen species and the high specific surface area of g-C3N4@PCN-224, this film still maintains 99.99% bactericidal efficacy against Escherichia coli and Staphylococcus aureus after 10 days of continuous bactericidal activity test. The results of cell and hemolysis experiments indicated that the film was safe and nontoxic and can effectively preserve fresh pork for 7 days. Moreover, the film also exhibited a recyclable and efficient killing activity. A strategy for achieving ultrapersistent freshness of perishable muscle food was provided.

A novel colorimetric and fluorometric dual-signal identification of organics and Baijiu based on nanozymes with peroxidase-like activity.

Nanozymes were nanomaterials with enzymatic properties. They had diverse functions, adjustable catalytic activity, high stability, and easy large-scale production, attracting interest in biosensing. However, nanozymes were scarcely applied in Baijiu identification. Herein, a colorimetric and fluorometric dual-signal determination mediated by a nanozyme-H2O2-TMB system was developed for the first time to identify organics and Baijiu. Since the diverse peroxidase-like activity of nanozymes, resulted in different degrees of oxidized TMB. Based on this, 21 organics were identified qualitatively and quantitatively by colorimetric method with a rapid response (<12 min), broad linearity (0.0005-35 mM), and low detection limits (a minimum of 30 nM for glutaric acids). Furthermore, the fluorometric method exhibited excellent potential for accurate determination of organics, with detection ranges of 2-200 µmol/L (LOD: 0.22 µmol/L) for l-ascorbic acid and 2-300 µmol/L (LOD: 0.59 µmol/L) for guaiacol. Finally, the sensor was successfully applied to identify fake Baijiu and Baijiu from 16 different brands.

Spinal anesthesia for L5-S1 interlaminar endoscopic lumbar discectomy: a retrospective study.

OBJECTIVE: This study aimed to report our experience with spinal anesthesia (SA) in patients undergoing L5-S1 interlaminar endoscopic lumbar discectomy (IELD) and clarify its advantages and disadvantages. METHODS: One hundred twelve patients who underwent IELD for an L5-S1 disc herniation under SA were retrospectively analyzed. SA with 0.5% ropivacaine was administered using a 27-gauge fine needle. Intraoperatively, the volume and level of SA, surgical time, blood loss, and cardiopulmonary complications were documented. Postoperative data was collected included the number of patients who ambulated on the day of surgery, incidence of complications and were then statistically analyzed. RESULTS: Analgesia was complete throughout the entire operation in all patients and no other adjuvant intraoperative analgesic drugs were needed. Mean visual analog scale scores for intraoperative and early postoperative (24 h) pain were 0 and 2.43 ± 1.66. SA was administered at the L3-4 interspace in 34 patients (30.4%) and the L2-3 interspace in 78 (69.6%). Administration was successful with the first attempt in all patients. Mean operation time was 70.12 ± 6.52 min. Mean intraoperative blood loss volume was 20.71 ± 5.26 ml. Ninety-eight patients ambulated on the same day as surgery. Mean length of hospital stay was 24.36 ± 3.64 h. Dural injury without damaging the nerve root occurred in one patient. One patient experienced recurrent disc herniation. Intraoperative hypotension and respiratory distress occurred in five (4.5%) and three (2.7%) patients, respectively. Three patients (2.7%) received postoperative analgesia therapy and two (1.8%) experienced nausea. Two patients (1.8%) developed urinary retention. Spinal headache, cauda equina syndrome, and neurotoxicity did not occur. CONCLUSION: SA can achieve satisfactory pain control for patients undergoing IELD with a low incidence of adverse events. SA may be a useful alternative to local and general anesthesia for IELD surgery. Future randomized controlled trials are warranted to investigate.

Next-generation meat preservation: integrating nano-natural substances to tackle hurdles and opportunities.

The increasing global meat demand raises concerns regarding the spoilage of meat caused by microbial invasion and oxidative decomposition. Natural substances, as a gift from nature to humanity, possess broad-spectrum bioactivity and have been utilized for meat preservation. However, their limited stability, solubility, and availability hinder their further development. To address this predicament, advanced organic nanocarriers provide an effective shelter for the formation of nano-natural substances (NNS). This review comprehensively presents various natural substances derived from plants, animals, and microorganisms, along with the challenges they face. Subsequently, the potential of organic nanocarriers is explored, highlighting their distinct features and applicability, in addressing these challenges. The review methodically examines the application of NNS in meat preservation, with a focus on their pathways of action and preservation mechanisms. Furthermore, the outlook and future trends for NNS applications in meat preservation are concluded. The theory and practice summary of NNS is expected to serve as a catalyst for advancements that enhance meat security, promote human health, and contribute to sustainable development.

Diversified organic nanocarriers conquer the limitations of natural substancesNNS based on organic nanocarriers are a reliable and health-promoting optionNNS can manifest their effectiveness through diverse pathways and mechanismsThe utilization of NNS in meat preservation represents a transformative strategy.

Recent advances in meat freshness "magnifier": fluorescence sensing.

To address the serious waste of meat resources and food safety problems caused by the decrease in meat freshness due to the action of microorganisms and enzymes, a low-cost, time-saving and high-efficiency freshness monitoring method is urgently needed. Fluorescence sensing could act as a "magnifier" for meat freshness monitoring due to its ability to sense characteristic signal produced by meat spoilage. Here, the magnification mechanism of meat freshness via sensing the water activity, adenosine triphosphate, hydrogen ion, total volatile basic nitrogen, hydrogen sulfide, bioamines was comprehensively analyzed. The existing "magnifier" forms including paper chips, films, labels, arrays, probes, and hydrogels as well as the application in livestock, poultry and aquatic meat freshness monitoring were reviewed. Future research directions involving innovation of principles, visualization and quantification capabilities for various meats freshness were provided. By critically evaluating the potential and limitations, efficient and reliable meat freshness monitoring strategies wish to be developed for the post-epidemic era.

Comparative Analysis of Volatile Flavor Compounds in Strongly Flavored Baijiu under Two Different Pit Cap Sealing Processes.

The solid-state fermentation process of strongly flavored Baijiu is complicated by the co-fermentation of many different microorganisms in the fermentation pools. The traditional fermentation pools of strong flavor Baijiu are sealed with mud, and this sealed-pit mud is not easy to maintain; therefore, the pit cap is prone to cracks and to caving in. The destruction of the sealed-pit mud may lead to instability in the composition and an abundance of microorganisms in the fermentation process that results in fluctuations of product quality. Thus, the production method of replacing the mud cap with a new steel cap is gradually attracting the attention of scientific and technical workers in the industry. However, so far, there have been relatively few reports on the use of steel lids for sealing pits for fermentation and brewing. In this study, the volatile flavor components of 270 Baijiu samples from mud-sealing and steel-sealing pits of a Chinese Baijiu distillery were studied qualitatively and quantitatively using Gas Chromatography-Mass Spectrometry (Abbreviated as GC-MS). Our statistical methods included Hierarchical Cluster Analysis (Abbreviated as HCA), Principal Component Analysis (Abbreviated as PCA), and Discriminant Analysis (Abbreviated as DA). A statistical analysis was carried out on the yield of strongly flavored Baijiu, and we made a comprehensive evaluation of the Baijiu produced under the two pit-sealing modes with regard to flavor and economic efficiency. The yield of strong flavored Baijiu was 6.7% higher with steel-sealing pits compared with mud-sealing pits. Cluster analysis categorized the strongly flavored Baijiu samples into two categories initially: (1) samples produced using mud-sealing pits and (2) samples using steel-sealing pits. Our analysis also indicated that the 28 compounds used for quantification were selected correctly. Surprising to the experimental staff, the overall score for the steel-sealing pits was greater than that of the mud-sealing pits based on PCA. Using DA, the prediction results were 100% accurate. In summary, through a comparative analysis of the flavor and yield, which are the two main factors that affect the quality of Baijiu in a distillery, and systematic combination at both experimental and theoretical levels, the differences between the Baijiu production by steel-sealing and the traditional mud-sealing were clear. Regardless of the impact of age, the detectable flavor components of Baijiu from the mud-steeling pits were very consistent with those of the steel-sealing pits in terms of richness or concentration. However, steel-sealing pits were significantly superior to mud-sealing pits with respect to output, consistency in quality, and cost (human and economic) savings.

Exploring the Successions in Microbial Community and Flavor of Daqu during Fermentation Produced by Different Pressing Patterns.

Daqu can be divided into artificially pressed daqu (A-Daqu) and mechanically pressed daqu (M-Daqu) based on pressing patterns. Here, we compared the discrepancies in physicochemical properties, volatile metabolites, and microbiota features between A-Daqu and M-Daqu during fermentation and further investigated the factors causing those differences. A-Daqu microbiota was characterized by six genera (e.g., Bacillus and Thermoactinomyces), while five genera (e.g., Bacillus and Thermomyces) dominated in M-Daqu. The flavor compounds analysis revealed that no obvious difference was observed in the type of esters between the two types of daqu, and M-Daqu was enriched with more alcohols. The factors related to differences between the two types of daqu were five genera (e.g., Hyphopichia). The functional prediction of microbial communities revealed that the functional discrepancies between the two types of daqu were mainly related to ethanol metabolism and 2,3-butanediol metabolism. This study provided a theoretical basis for understanding the heterogeneity of daqu due to the different pressing patterns.

Research on accurate non-contact temperature measurement method for telescope mirror.

Non-contact temperature measurement for a solar telescope mirror is critical for improving the mirror seeing and thermal deformation of solar telescopes, a long-standing challenge in astronomy. This challenge arises from the telescope mirror's inherent weak thermal radiation, often overwhelmed by reflected background radiations due to its high reflectivity. In this work, an infrared mirror thermometer (IMT) is equipped with a thermally-modulated reflector, and a measurement method based on an equation for extracting mirror radiation (EEMR) has been developed for probing the accurate radiation and temperature of the telescope mirror. Using this approach, we can extract the mirror radiation from the instrumental background radiation via the EEMR. This reflector has been designed to amplify the mirror radiation signal incident on the infrared sensor of IMT, while inhibiting the radiation noise from the ambient environment. In addition, we also propose a set of evaluation methods for IMT performance based on EEMR. The results reveal that the temperature measurement accuracy of IMT to the solar telescope mirror using this measurement method can be achieved better than ±0.15°C.

Study of transient behaviors of wavefront error caused by natural convection for improving mirror seeing.

Astronomical seeing parameters calculated based on the Kolmogorov turbulence model cannot fully evaluate the effect of the natural convection (NC) above a solar telescope mirror on the image quality, as the convective air motions and temperature variations of the NC are significantly different from the Kolmogorov turbulence. In this work, a new method based on the transient behaviors and frequency characteristics of NC-related wavefront error (WFE) are investigated in detail and used to evaluate the image quality degradation caused by a heated telescope mirror, aiming to make up for the deficiency of astronomical seeing parameters with the conventional method in evaluating the image quality degradation. Transient computational fluid dynamics (CFD) simulations and WFE calculations based on discrete sampling and ray segmentation are performed to quantitatively evaluate the transient behaviors of the NC-related WFE. It clearly exhibits apparent oscillatory characteristics, which are coupled by main oscillation with low frequencies and minor oscillation with high frequencies. Moreover, the generation mechanisms of two types of oscillations are studied. The conspicuous oscillation frequencies of the main oscillation caused by heated telescope mirrors with varying dimensions are primarily lower than 1 Hz, suggesting that active optics may be adopted to correct the main oscillation of NC-related WFE while the adaptive optics may correct the minor oscillation. Furthermore, a mathematical relationship between WFE, temperature rise, and mirror diameter is derived, revealing a significant correlation between WFE and mirror diameter. Our work suggests the transient NC-related WFE should be considered as one of the critical supplements to the mirror seeing evaluation.

Metaproteomic investigation of enzyme profile in daqu used for the production of Nongxiangxing baijiu.

Enzymes and microbiota in daqu are essential for the brewing of Nongxiangxing baijiu. Uncover the key enzymes and functional strains in daqu is beneficial to improve the flavor and quality of Nongxiangxing baijiu. In this study, metaproteome technology was employed to determine the enzyme profiles in Nongxiangxing daqu, and strains with high saccharification activity were screened and identified. 933 proteins were identified in daqu, of which 463 belonged to enzymes, including 140 oxidoreductases, 98 transferases, 91 hydrolases, 49 ligases, 41 lyases and 27 isomerases, and hydrolase is the enzyme with the highest abundance in baijiu brewing. Among hydrolases, a total of 36 carbohydrate metabolism-related enzymes (CMEs) were identified, and 12 of them were key enzymes related to glycoside hydrolysis. Four major glycoside hydrolysis enzymes glucoamylase (EC 3.2.1.3), glucan 1,4-alpha-glucosidase (EC 3.2.1.3), glucanase (EC 3.2.1.-) and ß-glucosidase (EC 3.2.1.21) were revealed, and their sources were Byssochlamys spectabilis, Lichtheimia ramosa and Thermoascus aurantiacus, respectively. Then, strains Aspergillus A2, A3, A7, Lichtheimia L1, L4, L5, and Saccharomycopsis S2, S4, S6 with high saccharifying enzyme-producing capacity were screened through culture-dependent approach. Resents presented in this study can further reveal the enzyme profiles and identify the main functional strains in daqu, which can provide theoretical support for the brewing of Nongxiangxing baijiu.

Biomechanics of open-globe injury: a review.

Open-globe injury is a common cause of blindness clinically caused by blunt trauma, sharp injury, or shock waves, characterised by rupture of the cornea or sclera and exposure of eye contents to the environment. It causes catastrophic damage to the globe, resulting in severe visual impairment and psychological trauma to the patient. Depending on the structure of the globe, the biomechanics causing ocular rupture can vary, and trauma to different parts of the globe can cause varying degrees of eye injury. The weak parts or parts of the eyeball in contact with foreign bodies rupture when biomechanics, such as external force, unit area impact energy, corneoscleral stress, and intraocular pressure exceed a certain value. Studying the biomechanics of open-globe injury and its influencing factors can provide a reference for eye-contact operations and the design of eye-protection devices. This review summarises the biomechanics of open-globe injury and the relevant factors.

A sensitive electrochemical sensor based on 3D porous melamine-doped rGO/MXene composite aerogel for the detection of heavy metal ions in the environment.

Herein, we developed a unique screen-printed carbon electrode (SPCE) with three-dimensional melamine-doped graphene oxide/MXene composite aerogel (3D MGMA) modification, which is used for the simultaneous and sensitive detection of three metal ions (Zn2+, Cd2+, and Pb2+) in the environment. A self-assembly method was used to fabricate 3D MXene aerogels based on MXene, graphene oxide (GO), and melamine. Notably, the network-like 3D structure combining 2D MXene and rGO sheets can provide a high ratio of surface area and enriched functional clusters, which are beneficial for improving the electrical conductivity and promoting the uptake of heavy metal ions. In the linear range of 3-900 µg L-1, the constructed innovative sensing platform can sensitively detect Zn2+, Cd2+, and Pb2+ simultaneously, with detection limits of 0.48 µg L-1,0.45 µg L-1 and 0.29 µg L-1 respectively. This work reflects precision and reliability in the detection of three water samples (tap water, Minzhu lake and Yangtze River) and four cereal samples (sorghum, rice, wheat and corn), proposing a novel strategy for monitoring heavy metal ions in the natural environment.

The microsphere of sodium alginate-chitosan-Pichia kudriavzevii enhanced esterase activity to increase the content of esters in Baijiu solid-state fermentation.

Pichia kudriavzevii was one of the important aroma-producing fungi in the solid-state fermentation of Baijiu, and immobilization was an effective strategy for improving microbial performance. Herein, P. kudriavzevii cells were immobilized in a gel network that crosslinked by chitosan and sodium alginate to form sodium alginate/chitosan-P. kudriavzevii microspheres (SA/CS-PMs). Their structural characteristics and formation processes were characterized by SEM and FT-IR. The effect of synthesis conditions on the performance of microspheres were determined by single-factor experiments. Under the optimal conditions, the SA/CS-PMs could increase the amylase activity of the fermentation broth by 57.18%, the esterase activity by 66.13%, the content of ester by 67.04%, and could be reused at least three times. Further research results indicated that the content of ester could be increased significantly in Baijiu solid-state fermentation with the SA/CS-PMs. In conclusion, the SA/CS-PMs could improve the ester production ability of P. kudriavzevii by increasing the esterase activity, which was a valuable exploration of directional biosynthesis and a feasible strategy to improve solid-state fermentation quality.

Butelase 1-Mediated Enzymatic Cyclization of Antimicrobial Peptides: Improvements on Stability and Bioactivity.

Antimicrobial peptides (AMPs) have broad-spectrum antibacterial properties and safety as food preservatives, whereas the stability and antibacterial activity require improvement. Here, the "head-to-tail" cyclization of linear AMP GKE was catalyzed by butelase 1, which resulted in an improved pronouncedly antibacterial effect. Cell morphology and propidium iodide uptake revealed that the increased membrane permeability was one of the bacteriostatic mechanisms of GKE and could be enhanced after cyclization. As cyclic GKE (cGKE) exhibited more stability than the linear counterpart under the microorganism culture environment, the increase in effective bacteriostatic concentration should be a reason for the superior antibacterial effect. Moreover, cGKE exhibited the ordered secondary structure, while GKE possessed a similar structure only in sodium dodecyl sulfate micelles. The structure was also beneficial to improve the antibacterial activity caused by the increased affinity of cGKE to the membranes. Overall, butelase 1-mediated cyclization is a promising strategy for enhancing the antibacterial activity of linear AMPs.

Green tea polyphenols bind to soy proteins and decrease the activity of soybean trypsin inhibitors (STIs) in heated soymilk.

The interaction between epigallocatechin gallate (EGCG) and soy proteins at room temperature (25 °C) and after heating at 100 and 121 °C, and their effects on the inactivation of soybean trypsin inhibitors (STIs) in soymilk were investigated. The results of the nitroblue tetrazolium (NBT) staining assay showed that soy proteins can covalently bind to EGCG. The α/α' and A subunits in heated soymilk preferred to bind to EGCG because of their soluble state. More thiols were trapped when EGCG was added before thermal processing, and the free amino groups were depleted more with EGCG addition after heating. Circular dichroism and fluorescence spectroscopy showed that EGCG addition before or after heating induced different secondary and tertiary structural changes for soy proteins. The exposed aromatic amino acids preferred to react with EGCG before protein aggregation in the heating process. The random coil of soymilk proteins increased more when EGCG was added in soymilk after heating, resulting in more disordered structures in protein conformation. The binding between EGCG and soy proteins promoted protein aggregation, which was confirmed by the particle size distribution and gel electrophoresis. The trypsin and chymotrypsin inhibitory activity (TIA and CIA) in soymilk significantly reduced to 693 U mL-1 and 613 U mL-1, respectively, under the conditions of 2 mM EGCG addition after 100 °C heating for 10 min (p < 0.05). Consequently, the influence of EGCG on STI inactivation in soymilk only worked when EGCG was added after heating.