Metabolic insights into the mechanism of soybean reddening during storage.

BACKGROUND: Soybean reddening during storage and transportation has caused great concern due to the serious economic loss. However, the mechanism of reddening has not been clearly elucidated. In this study, metabolomics was employed to investigate the reasons for soybean reddening during storage. RESULTS: The results of multivariate statistical analysis showed that the metabolite level of red soybean was significantly different from that of normal soybean. The differentially expressed metabolites were mainly enriched by biosynthesis of secondary metabolites and amino acid metabolism. Metabolism analysis showed that the biosynthesis of cyanidin and betalains was enhanced in reddening soybean. In addition, it was found that phenolic and flavonoid compounds decreased, while quinones, furans and 5-hydroxymethylfurfural increased in reddening soybeans compared to normal soybeans. CONCLUSION: The upregulation of cyanidin and betalains was the main reason for soybean reddening. Besides, the oxidation of phenols and flavonoids, as well as Maillard reaction, also contributed to the color change. © 2024 Society of Chemical Industry.

Microdroplet-enhanced chip platform for high-throughput immunotherapy marker screening from extracellular vesicle RNAs and membrane proteins.

Extracellular vesicles (EVs) are considered as promising candidates for predicting patients who respond to immunotherapy. Nevertheless, simultaneous detection of multiple EVs markers still presents significant technical challenges. In this work, we developed a high-throughput microdroplet-enhanced chip (MEC) platform, which utilizes thousands of individual microchambers (∼pL) as reactors, accelerating the detection efficiency of the CRISPR/Cas systems and increasing the sensitivity by up to 100-fold (aM level). Ten biomarkers (including 5 RNAs and 5 proteins) from patients' EVs are successfully detected on one chip, and the comprehensive markers show increased accuracy (AUC 0.911) than the individual marker for the efficacy prediction of immunotherapy. This platform provides a high-throughput yet sensitive strategy for screening immunotherapy markers in clinical.

Transcriptomics revealed the key molecular mechanisms of ofloxacin-induced hormesis in Chlorella pyrenoidosa at environmentally relevant concentration.

Emerging pollutants such as antibiotics have aroused great concern in recent years. However, the knowledge of low concentration-induced hormesis was not well understood. This study evaluated and quantified hormetic effects of ofloxacin on Chlorella pyrenoidosa. LogNormal model predicted the maximal non-effect concentration was 0.13 mg/L and 2.96 mg/L at 3 and 21 d, respectively. The sensitive alterations in chlorophyll fluorescence suggested PSII was the main target. Transcriptomics revealed ofloxacin inhibited genes related to photosynthetic system while the cyclic electron around PSI decreased the pH value in stroma side and stimulated photoprotection via up-regulating psbS. The stimulation in citrate cycle pathway met the urgent requirements of energy for DNA replication and repair. In addition, the negative feedback of G3P in glycolysis pathway inhibited Calvin cycle. The degradation products illustrated the occurrence of multiple detoxification mechanisms such as demethylation and ring-opening. The mobilization of cytochrome P450 generated the constant detoxication of ofloxacin while glutathione was consumptively involved in biological binding. This study provided new insights into the molecular mechanisms of antibiotic-induced hormesis in microalgae.

Isolation and Identification of Phosphate-solubilizing Bacteria in the Rhizosphere of Robinia pseudoacacia on the Loess Plateau and Verification of Phosphate Solubilization Capacity.

The Loess Plateau is one of the key areas for soil and water erosion control in China. Planting vegetation, such as Robinia pseudoacacia, is one of the mainstream methods to prevent soil and water erosion. However, the combination of abundant calcium ions and phosphate in the soil of the Loess Plateau limits the phosphorus nutrition of plants. In the present study, soil samples were collected under the R. pseudoacacia forest, from which two PSB strains with efficient phosphate solubilization capacities, named PSB2 and PSB7, were isolated and screened. The dissolved phosphate concentrations of their culture media were 9.68-fold and 11.61-fold higher, respectively, than that of the control group. After identification, PSB2 was classified as Pseudomonas and PSB7 as Inquilinus. This is the first time that Inquilinus has been isolated as a PSB from calcareous soil in the Loess Plateau. We then investigated the effects of different growth conditions on their phosphate solubilization capacities. Both strains effectively utilized glucose and ammonium nitrogen while maintaining high phosphate solubilization efficiency. In addition, PSB2 preferred to survive under neutral conditions and PSB7 under acidic conditions. Pot experiments indicated that the inoculation with PSB7 significantly increased the phosphorus content in the roots of R. pseudoacacia. These results imply the potential of this PSB as a phosphorus biofertilizer for R. pseudoacacia, which may be beneficial for soil and water management on the Loess Plateau.

Characteristics of Bacteria in Urine and Stones from Patients Treated with Percutaneous Nephrolithotomy and Association with Postoperative Infection.

Background: The purpose of this study was to identify bacterial differences between urine cultures (UC) and stone cultures (SC) in patients with complex kidney stones and to determine any correlation with post-percutaneous nephrolithotomy Systemic Inflammatory Response Syndrome (SIRS). Methods: Perioperative data of 1055 patients with complex kidney stones treated with first-stage Percutaneous Nephrolithotomy (PCNL) from September 2016 until September 2021 were included. Preoperative mid-stream urine samples and surgically obtained stone material were subjected to bacterial culture and antibiotic sensitivity tests. Preoperatively, antibiotic usage was determined by the UC or local bacterial resistance patterns. After PCNL treatment, antibiotic selection was guided by stone bacterial culture result and clinical symptoms. The effect of different preoperative antibiotic regimens based on urine cultures and postoperative antibiotic treatment based on stone cultures were assessed. Results: Positive stone cultures (SC+) were significantly more common than positive urine cultures (UC+) (31.9% vs 20.9%, p < 0.05). Escherichia coli (E. coli) was the most common uropathogen in both urine (54.3%) and stones (43.9%). The difference was statistically significant (p < 0.05). Moreover, UC+SC-, UC-SC+, UC+SC+, and preoperative serum creatinine were independent risk factors of postoperative SIRS. The incidence of SIRS in the UC+SC+ patients with different bacteria in stones and urine (51.6%) was higher than that in other culture groups. The antibiotic resistance of E. coli inside the stone was increased when prolonged preoperative antibiotics were administered to UC+ patients. Conclusion: The bacterial spectrum and positive outcome of culture in urine and stones were significantly different. The incidence of postoperative SIRS was highest in patients with UC+SC+ but with different bacteria strains. Prolonged pre-surgical antibiotic treatment apparently induced higher drug resistance for bacteria inside the stone.

LbCOPT1 is a copper transporter induced in Lycium barbarum mycorrhizal roots, which allows tobacco with improved growth and nutrient uptake.

KEY MESSAGE: Overexpressing the copper transporter LbCOPT1 leads to a notable increase in the abundance of mycorrhizal arbuscules that suggests the potential application of LbCOPT1 in breeding programs aimed at enhancing symbiotic nutrient uptake in Lycium barbarum L.

Preparation, pungency and bioactivity transduction of piperine from black pepper (Piper nigrum L.): A comprehensive review.

Piperine, derived from black pepper (Piper nigrum L.), is responsible for the pungent sensation. The diverse bioactivities of piperine underscores its promising potential as a functional food ingredient. This review presents a comprehensive overview of the research progress in extraction, synthesis, pungency transduction mechanism and bioactivities of piperine. Piperine can be extracted through various methods, such as traditional, modern, and innovative extraction techniques. Its synthesis mainly included both chemical and biosynthetic approaches. It exhibits a diverse range of bioactivities, including anticancer, anticonvulsant, antidepressant, anti-inflammatory, antioxidant, immunomodulatory, anti-obesity, neuroprotective, antidiabetic, hepatoprotective, and cardiovascular protective activities. Piperine can bind to TRPV1 receptor to elicit pungent sensation. Overall, the present review can provide a theoretical reference for advancing the potential application of piperine in the field of food science.

Insight into the multi-scale structure and retrogradation of corn starch by partial gelatinization synergizing with epicatechin/epigallocatechin gallate.

Starch retrogradation is of great importance to the quality of starch-based food. This study investigated the effect of partial gelatinization (PG) synergizing with polyphenol (epicatechin, EC; epigallocatechin gallate, EGCG) on the multi-scale structure and short/long-term retrogradation of corn starch (CS). The PG synergizing with EC/EGCG substantially suppressed the short/long-term retrogradation properties of CS. These could be confirmed by the decreased storage modulus and viscosity, the relative crystallinity (1.54%, 3.56%), and the retrogradation degree (9.99%, 20.18%) of CS during storage for 1, 14 days after PG synergizing with EGCG and EC, respectively. This is because PG treatment promoted the hydrogen bond interaction between disordered starch molecules and EC/EGCG. These were proven by the larger aggregation, more and brighter fluorescents, and the reduced long/short-range order structures in CS after PG synergizing with EC/EGCG. This study is helpful for the development of foods with enhanced nutrition and low-retrogradation.

Mechanisms of hormetic effects of ofloxacin on Chlorella pyrenoidosa under environmental-relevant concentration and long-term exposure.

Antibiotics are frequently detected in surface water and pose potential threats to organisms in aquatic ecosystem such as microalgae. The occurrence of biphasic dose responses raised the possibility of stimulation of microalgal biomass by antibiotics at environmental-relevant concentration and caused potential ecological risk such as algal bloom. However, the underlying mechanisms of low concentration-induced hormetic effects are not well understood. In this study, we evaluated the hormesis of ofloxacin on Chlorella pyrenoidosa under environmental-relevant concentration and long-term exposure. Results showed the hormetic effects of ofloxacin on cell density and carbon fixation rate (RC). The predicted maximum promotion was 17.45 % by 16.84 µg/L and 20.08 % by 15.78 µg/L at 21 d, respectively. The predicted maximum concentration of non-effect on cell density and RC at 21 d was 3.24 mg/L and 1.44 mg/L, respectively. Ofloxacin induced the mobilization of pigments and antioxidant enzymes to deal with oxidative stress. PCA analysis revealed Chl-a/Chl-b could act as a more sensitive biomarker under acute exposure while chlorophyll fluorescence parameters were in favor of monitoring long-term implication. The hormesis in increased secretion of extracellular organic matters was regarded as a defensive mechanism and accelerated indirect photodegradation of ofloxacin. Bioremoval was dominant and related to biomass accumulation in the total dissipation while abiotic removal appeared slight contributions. This study provided new insights into the understanding of hormesis of microalgae induced by antibiotics.

A CT-based integrated model for preoperative prediction of occult lymph node metastasis in early tongue cancer.

Background: Occult lymph node metastasis (OLNM) is an essential prognostic factor for early-stage tongue cancer (cT1-2N0M0) and a determinant of treatment decisions. Therefore, accurate prediction of OLNM can significantly impact the clinical management and outcomes of patients with tongue cancer. The aim of this study was to develop and validate a multiomics-based model to predict OLNM in patients with early-stage tongue cancer. Methods: The data of 125 patients diagnosed with early-stage tongue cancer (cT1-2N0M0) who underwent primary surgical treatment and elective neck dissection were retrospectively analyzed. A total of 100 patients were randomly assigned to the training set and 25 to the test set. The preoperative contrast-enhanced computed tomography (CT) and clinical data on these patients were collected. Radiomics features were extracted from the primary tumor as the region of interest (ROI) on CT images, and correlation analysis and the least absolute shrinkage and selection operator (LASSO) method were used to identify the most relevant features. A support vector machine (SVM) classifier was constructed and compared with other machine learning algorithms. With the same method, a clinical model was built and the peri-tumoral and intra-tumoral images were selected as the input for the deep learning model. The stacking ensemble technique was used to combine the multiple models. The predictive performance of the integrated model was evaluated for accuracy, sensitivity, specificity, and the area under the receiver operating characteristic curve (AUC-ROC), and compared with expert assessment. Internal validation was performed using a stratified five-fold cross-validation approach. Results: Of the 125 patients, 41 (32.8%) showed OLNM on postoperative pathological examination. The integrated model achieved higher predictive performance compared with the individual models, with an accuracy of 84%, a sensitivity of 100%, a specificity of 76.5%, and an AUC-ROC of 0.949 (95% CI [0.870-1.000]). In addition, the performance of the integrated model surpassed that of younger doctors and was comparable to the evaluation of experienced doctors. Conclusions: The multiomics-based model can accurately predict OLNM in patients with early-stage tongue cancer, and may serve as a valuable decision-making tool to determine the appropriate treatment and avoid unnecessary neck surgery in patients without OLNM.

The Ideal Proportion of the Auricle Exposure via a Morphometric Analysis in Asian Women.

BACKGROUND: Ears are an important aesthetic feature that is vital to the overall attractiveness of the face. Although there have been many studies on the aesthetics of the auricle, there is currently a lack of consensus on the ideal proportion of auricle exposure for Asian women in frontal view. OBJECTIVES: This study aimed to investigate ideal proportion of auricle exposure in Asian women. METHODS: An observational study was carried out on the photographs of 84 women on the list of the 100 most beautiful faces in Asia (published by TCC Asia in 2020). The proportion of the distance between the outer canthus and the outermost point of auricle to the distance between the inner canthus and the outermost point of auricle was calculated as the auricle exposure proportion. Evaluators were asked to rank a set of photographs of the volunteer with varying auricle exposure proportions from most attractive to least attractive. RESULTS: Measurements of the photographs of the 84 women showed a mean ear exposure proportion of 0.600. With 487 questionnaire responses received, the proportion of auricle exposure that the evaluators considered most attractive was 0.600. People with aesthetic experience considered 0.625 the most attractive proportion, while the general group considered 0.600 the most attractive. CONCLUSIONS: The ideal proportion of the auricle exposure for Asian women is in the range of 0.60-0.625, which may help surgeons reconstruct aesthetically pleasing ears. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Genetic and phenotypic profiling of single living circulating tumor cells from patients with microfluidics.

Accurate prediction of the efficacy of immunotherapy for cancer patients through the characterization of both genetic and phenotypic heterogeneity in individual patient cells holds great promise in informing targeted treatments, and ultimately in improving care pathways and clinical outcomes. Here, we describe the nanoplatform for interrogating living cell host-gene and (micro-)environment (NICHE) relationships, that integrates micro- and nanofluidics to enable highly efficient capture of circulating tumor cells (CTCs) from blood samples. The platform uses a unique nanopore-enhanced electrodelivery system that efficiently and rapidly integrates stable multichannel fluorescence probes into living CTCs for in situ quantification of target gene expression, while on-chip coculturing of CTCs with immune cells allows for the real-time correlative quantification of their phenotypic heterogeneities in response to immune checkpoint inhibitors (ICI). The NICHE microfluidic device provides a unique ability to perform both gene expression and phenotypic analysis on the same single cells in situ, allowing us to generate a predictive index for screening patients who could benefit from ICI. This index, which simultaneously integrates the heterogeneity of single cellular responses for both gene expression and phenotype, was validated by clinically tracing 80 non-small cell lung cancer patients, demonstrating significantly higher AUC (area under the curve) (0.906) than current clinical reference for immunotherapy prediction.

Study on peanut protein oxidation and metabolomics/proteomics analysis of peanut response under hypoxic/re-aeration storage.

To better understand the effect of oxygen levels in the storage environment on peanut protein oxidation and explore the mechanism, the functional properties and the oxidation degree of peanut proteins extracted from peanuts under conventional storage (CS), nitrogen modified atmosphere storage (NS, hypoxic) and re-aeration storage (RS) were investigated. Metabolomics and proteomics were employed to analyze peanut's response to hypoxic/re-aeration storage environment. The results showed that NS retarded the decline of the functional properties and the oxidation of peanut proteins, while the process were accelerated after re-aeration. That was the result of the metabolic changes of peanuts under different storage environments. The omics results presented the decreased (NS)/increased (RS) levels of the antioxidant-related proteins acetaldehyde dehydrogenase and glutathione S-transferase, and the inhibition (NS)/activation (RS) of metabolic pathways such as the TCA cycle and the pentose phosphate pathway. This study provided a reference for the re-aeration storage of other agricultural products.

EMX2 inhibits clear cell renal cell carcinoma progress via modulating Akt/FOXO3a pathway.

Empty spiracles homeobox 2 (EMX2) is initially identified as a key transcription factor that plays an essential role in the regulation of neuronal development and some brain disorders. Recently, several studies emphasized that EMX2 could as a tumor suppressor, but its role in human clear cell renal cell carcinoma (ccRCC) remains unclear. In the present study, we investigated the role and underlying mechanism of EMX2 in the regulation of ccRCC progress. Our results demonstrated that EMX2 expression was markedly decreased in ccRCC tissues and cell lines, and low EMX2 expression predicted the poor prognosis of ccRCC patients. In addition, forced expression of EMX2 significantly inhibited the cell growth, migration, and invasion in vitro, as well as ccRCC tumor growth in nude mice, via, at least in part, regulating Akt/FOXO3a pathway. In detail, EMX2 could attenuate the phosphorylation levels of Akt and FOXO3a, and increase FOXO3a expression without affecting total Akt expression in vivo and in vitro. Meanwhile, shRNA-mediated knockdown of FOXO3a expression could obviously attenuate the effects of EMX2 on cell growth, migration, invasion, and tumor growth. Furthermore, EMX2 could significantly attenuate the interaction between Akt and FOXO3a. Taken together, our results demonstrated that EMX2 could inhibit ccRCC progress through, at least in part, modulating Akt/FOXO3a signaling pathway, thus representing a novel role and underlying mechanism of EMX2 in the regulation of ccRCC progress.

Physiological-biochemical responses and transcriptomic analysis reveal the effects and mechanisms of sulfamethoxazole on the carbon fixation function of Chlorella pyrenoidosa.

The occurrence of sulfamethoxazole (SMX) is characterized by low concentration and pseudo-persistence. However, the toxic effects and mechanisms of SMX, especially for low concentration and long-term exposure, are still not clear. This study investigated the effects and mechanisms of SMX on carbon fixation-related biological processes of Chlorella pyrenoidosa at population, physiological-biochemical, and transcriptional levels. Results showed that 1-1000 µg/L SMX significantly inhibited the dry weight and carbon fixation rate of C. pyrenoidosa during 21 d. The upregulation of superoxide dismutase (SOD) and catalase (CAT) activities, as well as the accumulation of malondialdehyde (MDA) demonstrated that SMX posed oxidative damage to C. pyrenoidosa. SMX inhibited the activity of carbonic anhydrase (CA), and consequently stimulated the activity of Rubisco. Principal component analysis (PCA) revealed that SMX concentration was positively correlated with Rubisco and CAT while exposure time was negatively correlated with CA. Transcriptional analysis showed that the synthesis of chlorophyll-a was stabilized by regulating the diversion of protoporphyrin IX and the chlorophyll cycle. Meanwhile, multiple CO2 compensation mechanisms, including photorespiratory, C4-like CO2 compensation and purine metabolism pathways were triggered in response to the CO2 requirements of Rubisco. This study provides a scientific basis for the comprehensive assessment of the ecological risk of SMX.

Histone deacetylase 6 suppression of renal tubular epithelial cell promotes interstitial mineral deposition via alpha-tubulin acetylation.

Randall's plaque (RP) is derived from interstitial mineral deposition and is highly prevalent in renal calcium oxalate (CaOx) stone disease, which is predictive of recurrence. This study shows that histone deacetylase 6 (HDAC6) levels are suppressed in renal tubular epithelial cells in RP samples, in kidney tissues of hyperoxaluria rats, and in hyper-oxalate-treated or mineralized cultured renal tubular epithelial (MDCK) cells in vitro. Mineral deposition in MDCK cells was exacerbated by HDAC6 inhibition but alleviated by HDAC6 overexpression. Surprisingly, the expression of some osteogenic-associated proteins, were not increased along with the increasing of mineral deposition, and result of single-cell RNA sequencing of renal papillae samples revealed that epithelial cells possess lower calcific activity, suggesting that osteogenic-transdifferentiation may not have actually occurred in tubular epithelial cells despite mineral deposition. The initial mineral depositions facilitated by HDAC6 inhibitor were localized in extracellular dome rather than inside the cells, moreover, suppression of HDAC6 significantly increased the calcium content of co-cultured renal interstitial fibroblasts (NRK49F) and enhanced mineral deposition of indirectly co-cultured NRK49F cells, suggesting that HDAC6 may influence trans-MDCK monolayer secretion of mineral. Further experiments revealed that this regulatory role was partially alpha-tubulinLys40 acetylation dependent. Collectively, these results suggest that hyper-oxalate exposure led to HDAC6 suppression in renal tubular epithelial cells, which may contribute to interstitial mineral deposition by promoting alpha-tubulinLys40 acetylation. Therapeutic agents that influence HDAC6 activity may be beneficial in preventing RP and CaOx stone formation.

Scoparone attenuates glioma progression and improves the toxicity of temozolomide by suppressing RhoA/ROCK1 signaling.

BACKGROUND: Glioma, a type of malignant brain tumor, has become a challenging health issue globally in recent years. METHODS: In this study, we investigated the potential therapeutic role of scoparone in glioma and the underlying mechanism. Initially, transcriptome sequencing was conducted to identify genes that exhibited differential expression in glioma cells treated with scoparone compared to untreated cells. Subsequently, the impact of scoparone on the proliferation, migration, and invasion of glioma cells was assessed in vitro using a range of assays including cell viability, colony formation, wound healing, and transwell assays. Moreover, the apoptotic effects of scoparone on glioma cells were evaluated through flow cytometry and western blot analysis. Furthermore, we established a glioma xenograft mouse model to assess the in vivo antitumor activity of scoparone. Lastly, by integrating transcriptome analysis, we endeavored to unravel the molecular mechanisms underlying the observed antitumor effects of scoparone by examining the expression levels of RhoA/ROCK1 signaling pathway components using western blot analysis and qRT-PCR. RESULTS: Our transcriptome sequencing results revealed that scoparone significantly downregulated RhoA/ROCK1 signaling in glioma cells. Furthermore, scoparone treatment inhibited glioma cell proliferation, migration, and invasion, and promoted cell apoptosis in vitro. Moreover, scoparone reduced tumor growth and prolonged survival in a glioma xenograft mouse model, and improved the toxicity of temozolomide. Finally, our results showed that the antitumor effects of scoparone were mediated by the suppression of RhoA/ROCK1 signaling. CONCLUSION: Scoparone could be a promising therapeutic agent for glioma by suppressing RhoA/ROCK1 signaling. These findings pave the way for future research endeavors aimed at the development and optimization of scoparone-based therapeutic strategies.

The Auricular Injection Technique: A New Treatment for Lying Ear.

SUMMARY: Lying-ear deformity refers to an auricle that bends backward excessively, is excessively folded against the head, and has a very prominent antihelix. It usually requires experienced surgeons to perform surgical treatment and a prolonged postoperative recovery process. This article proposes a simple and effective hyaluronic acid injection technique that significantly improves the shape of the outer ear and enhances perceived facial aesthetics. Twenty patients underwent treatment with multiple injections. Measure-related parameters were used to evaluate the postoperative effect, and the results were graded using a visual analog scale. Interrater reliability among graders was evaluated using intraclass correlation coefficients. After treatment, no serious complications, such as infection or embolism, occurred. Six months after the procedure, the average auriculocephalic angle increased from 25.11 ± 9.46 to 32.72 ± 8.29 degrees, the average conchoscaphal angle increased from 87.69 ± 9.06 to 95.94 ± 7.11 degrees, and patients' average visual analog scale score increased from 4.40 ± 1.14 to 8.57 ± 0.68. Interrater reliability was fair to good for visual analog scale before injection and 6 months after injection (intraclass correlation coefficients, 0.49 and 0.45, respectively; both P < 0.001). The patients were satisfied with the injection process and results. This injection protocol improved the shape of the outer ear, resulting in excellent postoperative outcomes.

Peeling mechanism of tomato induced by HHAIB: Microscopic, ultrastructure, chemical, physical and mechanical properties perspectives.

In order to better manage the peeling degree and avoid unnecessary losses, the current work aimed to explore the peeling mechanism of a novel peeling technology, high-humidity hot air impingement blanching (HHAIB). The relationships between HHAIB peeling performance and the changes in skin temperature, skin structure, water state, pectin fractions content, and skin mechanical properties of tomatoes were analyzed. Results showed, after HHAIB treatment, the epicuticular wax was disrupted, the skin exhibited more and longer random cracks, the degradation of inner skin tissue was observed by transmission electron microscopy, the free water percentage increased resulting in water loss in the whole tomato, the water-soluble pectin contents decreased in tomato fleshes, while the contents of chelate-soluble pectin and sodium-carbonate-soluble pectin increased. HHAIB heating reduced the elongation at break, and increased Young's Modulus of tomato peel. This study revealed the HHAIB peeling mechanism and provided new insights for developing HHAIB peeling technology.