IntroductionAs COVID-19 shifts from pandemic urgency to endemic management, healthcare systems are faced with the evolving challenge of providing optimized care and adept resource allocation in this evolving landscape of the disease. However, the timely management and accurate assessment of disease severity remains a cornerstone of effective treatment. This study presents a pioneering scoring system, based on the primary chest CT scan findings, to predict patient outcomes and to equip clinicians with a tool that can expedite decision-making.MethodA retrospective cohort study was conducted involving 406 confirmed COVID-19 cases referred to two of our hospitals in Tehran, between February and April 2020. Radiographic and CT scan data were sourced from the imaging archive system and evaluated by a certified radiologist. We devised distinct severity scores for CT findings, demographic factors, and clinical indicators. These were synthesized into a comprehensive severity score to forecast critical patient outcomes, such as mortality, ICU admission, intubation, or extended hospitalization. Of the total cases, 161 (39.7%) were classified as severe, while 245 (60%) fell into the low or moderate severity category.ResultsThe mean score of demographic, CT scan, and clinical characteristics was significantly higher for those in the severe COVID-19 than the non-severe group. The cutoff score for predicting the outcomes in COVID-19 patients for demographic, clinical, and chest CT scan factors was 2.5, 9.5, and 8.5, respectively. Multivariate analysis indicated that each unit increase in these scores elevated the odds of fatal outcomes by 24%, 2.8%, and 12%, respectively. Then, using the comprehensive severity score, which is the sum of the above scores, we further predicted the disease severity.ConclusionThe findings suggest that our innovative scoring system, based on initial chest CT scan findings, serves as a robust predictor of COVID-19 outcomes. In late December 2019, Wuhan, China, became the epicenter for a surge of severe pneumonia cases, later attributed to the SARS-CoV-2 virus, leading to the identification of COVID-19 as a distinct disease. By February 2020, the World Health Organization had escalated its status to that of a global pandemic, signaling its far-reaching impact and the urgent need for international attention. Since then, the pandemic has proliferated globally, resulting in substantial morbidity and mortality . Most COConsidering the risks that COVID-19 poses to the individual, which is seen only in a small fraction of people with severe COVID-19, it warrants having a reliable predictive method for COVID-19 severity to distinguish that susceptible minority from the majority with benign disease course and to protect them against critical consequences. This risk stratification, which forecasts the patient’s condition and disease progression, assists healthcare front liners in providing more tailored medical management and earlier therapeutic intervention for high-risk patients.This study introduces a novel scoring system, amalgamating chest CT scan findings with demographic and clinical variables, aimed at predicting COVID-19 critical outcomes in patients. By correlating an encompassing severity score with pivotal clinical findings, we aspire to provide clinicians with a tool to promptly assess disease severity and manage patient care and resources efficiently, potentially shaping adaptive strategies in the face of an evolving healthcare landscape.Ethical approval, data collection, and image acquisitionThe Ethics Committee of Iran University of Medical Science (IUMS) approved this retrospective cohort study (IR.IUMS.FMD.REC.1400.467). Data were collected from 425 medical records from COVID-19-confirmed cases referred to Hazrate Rasool-e-Akram and Firoozabadi hospitals affiliated to IUMS, between February 2020 and April 2020. Four trained medical researchers evaluated data using a checklist containing demographic, clinical, and radiologic characteristics. Clinical and demographic variables were obtained from clinical records, and in case of possible missing data, follow-up phone calls were scheduled.Chest CT scans were performed utilizing a standard protocol across two medical facilities to maintain consistency in the data acquisition process. The CT images were obtained using a Siemens SOMATOM device , and all scans were executed with patients in a supine position and preferred during end inspiration without the administration of contrast media. The commonly used scan parameters were as follows: tube voltage of 120 kVp, automatic tube current modulation (30-70 mAs), pitch of 1.2, and slice thickness of 5 mm. Image reconstruction was performed with a standard kernel, and the resulting images were viewed at window settings optimized for lung parenchyma . To minimize variability, a single board-certified radiologist (E.Z.), blinded to the clinical details and outcomes, evaluated the scans and assigned severity scores based on the extent of lung involvement, adhering to the predefined scoring criteria. All digital image data were stored and analyzed using the picture archiving and communication system (PACS), ensuring a structured and standardized evaluation.Inclusion and exclusion criteriaInclusion criteria encompassed confirmed COVID-19 cases, validated either through chest CT scans and clinical symptoms or through RT-PCR, only including cases aged 18 years or older . The excDevelopment of scoring toolsPatients were assessed for COVID-19 severity according to three checklists, which were completed based on demographic factors in Table Subsequently, considering the disease progression, we stratified our patients into two outcome groups of severe COVID-19 and non-severe COVID-19 on the basis of occurrence of death, need for intubation, ICU admission, or prolonged hospital stay.The radiologist estimated the CT severity score according to the American College of Radiology (ACR) criteria. The severity of the chest CT scan and its quantitative trend was reported by introducing a specific definition of CT score (which ranged from 0 to 25) and divided into four groups. Using this simple score, the range of lung involvement was classified by a radiologist from 0 (0% or nothing), 1 (1-5% or at least), 2 (6-25% or low), 3 (26-49% or moderate), 4 (50-75% or severe), and 5 (75% ≤ extended) . We intrThe design of this decision-making tool is an innovative action and is not based on a specific reference. This comprehensive score was constructed through careful observation of the behaviors of experts, including emergency, lung, and infectious disease specialists, in making decisions in the field upon stratifying high-risk patients. Additionally, in line with the literature review, the weight of some disease symptoms in prognosis was obtained and was integrated as a criterion in this tool.Here, a chest CT scan is performed at the physician's discretion and maintains a pivotal role in the evaluation for subsequent follow-ups. The decision for hospitalization is made based on the severity of chest CT scan results in C5 criteria. However, C3 and C4 criteria are in the gray zone, meaning the severity of clinical or chest CT scan results is not merely enough to decide on hospitalization. Hence, we postulated that, by a combination of these three different criteria, a predictive measure for hospitalization could be generated in which higher scores denote higher severity of the disease.Statistical analysisDescriptive analyses were applied for demographic, clinical, and chest CT severity score data. An Independent T-test was used to compare means in each group. Pearson's correlation and chi-square test were used for assessing categorical variables. Logistic regression was used for assessing independent predictor variables and controlling confounders. The receiver operating characteristic (ROC) curve was applied to estimate the chest CT scan severity cutoff score in predicting the outcomes in patients. P value < 0.05 was considered statistically significant. All analyses were performed using Statistical Product and Service Solutions (SPSS) .Our studied population included 406 patients, comprising 172 (42.3%) women and 234 (57.7%) men. The mean ± SD age of patients was 61.6 ± 17.2 and ranged from 18 to 98. The mean ± SD of BMI was 25.9 ± 7.9. Considering management settings, 366 (90.1%) required hospitalization, and the other 40 (9.9%) were treated as outpatient. Finally, 161 (39.7%) were stratified as the "severe COVID-19" group, and 245 (60.3%) were in the "non-severe COVID-19" group.2 saturation, psychological and physical disorders, cardiac disorder, liver cirrhosis, active cancer, and having chemotherapy in the last six months were significantly associated with severe outcome (p < 0.05). There are no significant differences in other demographic and clinical variables between the two groups (p > 0.05)  Table .The association of outcomes with demographic and clinical characteristics scoresA comparison of the median overall score of demographic and clinical characteristics indicated that the mean overall score of demographic factors was significantly higher for people in the severe COVID-19 group than that of the non-severe group . Moreover, the mean overall score of clinical criteria was significantly higher in the severe COVID-19 group than in the non-severe group . A comparison of chest CT scan outcomes revealed that the number of people with > 50% lung involvement was significantly higher in the severe COVID-19 group as 39 (24.3%) vs 14 (5.7%), respectively (p = 0.001). The distribution of different degrees of lung involvement between the two groups is depicted in Table According to the comprehensive CT score, the number of people that were categorized as the severe COVID-19 group was significantly higher in each of C1-C5 sub-groups  Table .The results of multivariate logistic regression analyses showed that hospital stay duration was predicted with C1, which was statistically significant. In other words, the odds of prolonged hospital stay duration were significantly higher in people with C1 level than the others , and there was no significant association between C1 and the other outcomes. Being in C2 was anticipated intubation, being in C3 was projected with prolonged hospitalization and high odds of death (p > 0.05), and being in C4 was expected with the odds of intubation and death while having no significant association with the other outcomes (p > 0.05). Additionally, being in C5 predicted death, intubation, and ICU admission, and death with OR=2.25 was the most important predicting factor at this level , clinical , and chest CT scan  scores was correlated with higher odds of death .Based on ROC curve analysis results Figure , all thrFrom the earliest emergence of SARS-CoV-2 infection in Wuhan, China, multiple severe forms of the disease have been delineated ,9-11. AsThis study revealed that the chest CT scan severity score and the proposed clinical and demographic scales served as reliable predictors for COVID-19 severity. The highness of the mentioned scores compared to the cutoff determined for them forecast a higher chance of death, hospitalization, ventilator support, and ICU admission. The odds of long hospitalization were significantly higher for people at the C1 level than the others. Being at the C2 level was predicted with intubation. Being in C3 was anticipated with death and prolonged hospitalization. C4 level foretold intubation and a higher probability of death. At the C5 level, the probability of death, intubation, and ICU admission was increased. In a study by Aziz-Ahari et al.  on a simWang et al.  conducteLi et al.  conducteThe major drawback to previous studies is the limitation of factors considered in predictive criteria. By the inclusion of multiple factors and proposing three distinct criteria, we were able to assess the weight of similar factors combined together and independent from other criteria. This allows us to take adept preventive measures while making policies for disease prevention and to mitigate mortality and morbidity of this infection. Importantly, the most unique feature of this study is conferring numeric scoring to the proposed multi-disciplinary criteria. With the implementation of the confirmed cutoff points of scoring scales, we could carry out quantitative risk stratification for patients at the initial visit to the emergency department and prioritize healthcare resources for cases at higher risk to improve overall prognosis. This is highly crucial, particularly in periods of disease surge, which engenders catastrophic shortages of health service resources, a phenomenon observed recurrently in the COVID-19 pandemic.In this study, a more comprehensive scoring method based on the first chest CT scan, demographic factors comprising old age, smoking, comorbidity like diabetes, clinical characteristics, and chest CT scan results such as the severity of lung involvement, has been proposed, which was proven to maintain high value in predicting several outcomes , and its predicting power was confirmed by ROC curve. Hence, we constructed an exhaustive forecast method so that no study has been carried out with such magnitude.Limitations and strengths of the studyOur study presents both notable strengths and limitations that warrant discussion. A primary limitation lies in its retrospective design, which inherently faces challenges, such as incomplete records. Such gaps in data could potentially impact the study's outcomes, suggesting that a prospective approach would yield more accurate results. Conversely, a significant strength of our research is the implementation of a comprehensive scoring system that integrates a wide array of factors - demographic, clinical, and radiographic - to predict patient outcomes, enhancing the study's robustness and applicability. Additionally, the validity of our prognostic tool may vary in subsequent pandemic peaks due to evolving viral strains and clinical practices, necessitating further validation during diverse pandemic phases.Even though the radiologist estimated the CT severity score based on defined criteria, we admit that radiologic interpretation inevitably incorporates an element of subjectivity, which could introduce variability into the results. Different radiologists may interpret images with subtle differences, and while our scoring system provides a structured framework, inherent subjectivity may still impact scoring, especially in borderline cases. This potential for variability and its subsequent influence on the study outcomes warrant acknowledgment as a limitation. One way to overcome such limitations is the incorporation of artificial intelligence-based image analysis to mitigate such subjectivity, ensuring even greater consistency and reliability in data interpretation.Our study indicated that the total number of demographic, clinical, and radiologic characteristics are independently related to severe forms of COVID-19. The scoring method based on the first chest CT scan (SMBF-CT) scale, which is calculated based on demographic, clinical, and radiographic findings of patients, was proven to predict the outcomes of COVID-19 patients and is useful for treatment decision-making. Thus, utilizing SMBF-CT can effectively detect disease earlier and decrease the severe outcomes and burden of disease.


In this study, we evaluated the role of a multidisciplinary team (MDT) in clinical practice for cervical cancer by analyzing the development of a single-case multidisciplinary consultation for cervical cancer.Patients in MDT consultations for cervical cancer were retrospectively analyzed for clinical information, decision content of MDT discussion, implementation, and follow-up results.P <0.05). MDT decisions were correlated with age; the younger the patients, the higher the implementation efficiency (P <0.05). The difference between MDT expectation in all implementation and partial implementation and age was statistically significant (P <0.05). No significant difference was found between age and MDT expectation in all not fully implemented decisions (P >0.05). Some decisions were not fully implemented due to economic status and fear of certain treatments of the patient.Of the 392 patients who met the inclusion criteria, 359 had a first episode, of which 284 were stage IA-IIA2 (79.11%) and 75 were stage IIB-IVB (20.89%). Of these 392, 33 had a recurrence (8.42%). A total of 416 cases were analyzed, and neoadjuvant chemotherapy with surgery was recommended in 43 cases, of which 40 cases were implemented, and 36 of the 40 achieved the expected outcome. Surgical treatment was recommended in 241 cases, of which 226 underwent surgery, and 215 of them achieved the expected outcome. Radiotherapy was recommended in 31 cases, of which 26 cases underwent it, and 22 of them achieved the expected efficacy. Concurrent chemoradiotherapy was recommended in 57 cases, of which 49 underwent it, and 39 of them achieved the expected efficacy. Other treatments were recommended in 44 cases, of which 23 cases were implemented, and 10 of them achieved the expected efficacy, with statistically significant differences compared with cases without implementation (The MDT plays an important role in clinical practice such as clinical staging, treatment plan, and the complete treatment management of patients with cervical cancer, which can significantly improve the near-term treatment effect, whereas its effect on a long-term prognosis needs further clinical observation and active exploration. Cervical cancer is the most common malignant tumor in gynecology. Each year, more than half a million women are diagnosed with cervical cancer, and the disease results in over 3,00,000 deaths worldwide. More than 85% of new cases and approximately 90% of deaths occur in developing countries , 2. WhetMultidisciplinary collaboration for cervical cancer treatment needs to integrate the clinical knowledge, skills, and experience of all relevant departments in a hospital or relevant multidisciplinary team (MDT) models in higher-level hospitals according to a patient’s physical and psychological conditions, pathological type, clinical stage, and the developmental tendency for high-quality diagnosis and a precise individualized treatment plan based on evidence-based medicine to achieve the best treatment outcome that is economical and can maximize the survival rate and quality of life , 13. ForThe present study was approved by the Ethical Committee of Ganzhou Cancer Hospital (2023002), and written informed consent was obtained from participants. The study was performed in accordance with the principles of the Declaration of Helsinki regarding research involving human subjects. Each patient provided written informed consent to participate after the nature of the study was explained to them.A retrospective analysis was performed on the development of multidisciplinary consultation for patients with cervical cancer who fulfilled the inclusion criteria. The role of MDT in the clinical practice of cervical cancer was evaluated.Patient inclusion criteria, patients with (i) pathologically confirmed cervical cancer; (ii) first diagnosis or previous treatment; (iii) at least one MDT discussion; and (iv) complete clinical data were included in the study. Patient exclusion criteria, (i) patients with cervical cancer who did not participate in MDT discussions; (ii) patients with incomplete primary study information; and (iii) patients who were lost to follow-ups were excluded from the study.Ganzhou Cancer Center/Ganzhou Cancer Hospital in China is a large-scale, tertiary-level, A-class specialized oncology hospital. In total, 392 patients with cervical cancer from the Department of Gynecology and Oncology of Ganzhou Cancer Center/Ganzhou Cancer Hospital from January 2017 to December 2020 who met the inclusion criteria were selected. We included 83 cases (21.17%) from 2017, 95 cases (24.24%) from 2018, 104 cases (26.53%) from 2019, and 110 cases (28.06%) from 2020. with 359 cases of first presentation, including 284 cases of stage IA-IIA2, 75 cases of stage IIB-IVB, and 33 cases of recurrence. The age ranged from 23 to 70 years, with a median age of 52.46 years. A total of 416 MDT discussions were conducted in all patients Table 1.Since its establishment in 2012, the cervical cancer MDT treatment at the Ganzhou Cancer Center has gradually improved and developed to maturity by 2016. The MDT consisted of the Department of Gynecologic Oncology as the core, the Radiotherapy Center and Chemotherapy Center as the backbone, along with the Departments of Urology, Colorectal Medicine, Critical Care Medicine, Anesthesiology, Imaging, Pathology, and Nutrition. Patients are admitted to the hospital after the suspicion or diagnosis of cervical cancer. All tests are performed to clarify the diagnosis; patients’ conditions, the MDT treatment concept, and its advantages are explained clearly to these patients and families, and an attending physician submits an MDT discussion after consent. An expert group further discusses these conditions, a chief specialist summarizes and proposes a treatment strategy, a secretary maintains a written record, and the secretary and attending physician jointly inform the patients and families of the MDT consultation decision Figure 2The MDT meeting is usually held in the conference room of the Department of Gynecologic Oncology or the hospital teleconsultation center and is attended by members of the MDT specialists, new doctors, and students from the gynecologic oncology department, chemotherapy center, and radiotherapy center. They discuss two to three patients at a time during 1–1.5 hours. After creating the treatment plan, the Department of Gynecologic Oncology, the Radiotherapy Center, and the Chemotherapy Center are responsible for its implementation.The 392 patients were followed up until June 2022; the follow-up period was of 7–65 months, and the mean follow-up period was of 25.6 months. The mean follow-up time was 29.6 months for the 20–40 age group, 28.4 months for the 41–50 age group, 27.1 months for the 51–60 age group, and 18.2 months for the 61–70 age group. The patients were followed up by the attending physician, MDT secretary, and hospital follow-up center according to the principles of oncology follow-up. The outpatient follow-up was performed by the attending physician, whereas the telephone follow-up was performed by the MDT secretary and hospital follow-up center. All MDT consultation decisions completed are “fully implemented”, those partially completed are “partially implemented”, and those not implemented are “not implemented”. The implementation status of the MDT consultation decision for each patient was analyzed, and the implementation effect was recorded.Implementation outcomes were classified as “meeting expectations” and “not meeting expectations”. Expectations were met when the diagnosis was confirmed by MDT discussion, radical resection was achieved by surgery, radical treatment was achieved by radiotherapy, and symptom improvement or disease control was achieved by palliative care. The “not meeting expectations” outcomes included no confirmed diagnosis after MDT discussion, tumor residual after surgery or radiotherapy, and failure to improve symptoms or prolong survival after palliative care.2 test was performed to compare statistical data between the groups, and P < 0.05 was considered statistically significant.Statistical analysis was performed using IBM SPSS Statistics for Windows, version 21.0. Data are expressed as percentages. A hypothesis testing model was constructed for classified data. The χThe MDT consultation was discussed in 416 cases for 392 patients, of which 16 cases attended the consultation twice and four cases attended thrice, and repeat MDT consultations accounted for 5.1%. All the 416 MDT consultations involved diagnosis and treatment, and patients attended MDT consultations to know accurate cancer staging and treatment strategies. Among them, neoadjuvant chemotherapy with surgery was recommended in 43 cases (10.34%), surgery was recommended in 241 cases (57.93%), radiotherapy was recommended in 31 cases (7.45%), simultaneous radiotherapy and chemotherapy were recommended in 57 cases (13.70%), and other treatments were recommended in 44 cases (10.58%). Chemotherapy was recommended in 15 cases (3.61%), chemotherapy with targeted therapy was recommended in 13 cases (3.13%), chemotherapy with immunotherapy was recommended in 11 cases (2.64%), and targeted therapy with immunotherapy was recommended in five cases (1.20%) for other treatments 3. Posto2 = 11.2286, P = 0.0106)  were “fully implemented”, 81 (19.47%) were “partially implemented”, and 52 (12.50%) were “not fully implemented” Table 4.2 = 29.4569, P = 0.0000)    of full and partial implementation, 325 (89.29%) cases met MDT expectations. The expected efficiency of full implementation was 93.99% (266/283), and the expected efficiency of partial implementation was 72.84% (59/81), with a statistically significant difference (χ 0.0000) Table 8.Patients undergoing monotherapy require multiple referrals or consultations, which can be time-consuming and result in poor adherence to treatment. A treatment model via an MDT integrates the advantages of various related disciplines to develop an individualized, high-quality, and precise treatment plan, which plays an important role in improving the survival rate, quality of life, and prognosis of patients –15. In tP < 0.05). Of note, 16.59% of patients with internal iliac vein injury, bowel injury, bladder injury, ureteral injury, and intraoperative placement of ureteral stents were joint surgeries between two specialties with adequate preoperative evaluation. Urology was the specialty with the highest cooperation with cervical cancer surgery. The decisions of all 416 cases involved diagnosis plus treatment, and treatment decisions included neoadjuvant chemotherapy plus surgery, surgery, radiotherapy, concurrent chemoradiotherapy, and other treatments. Other treatments included chemotherapy, chemotherapy plus targeted therapy, chemotherapy plus immunotherapy, and targeted plus immunotherapy. All postoperative outcomes of surgical patients were discussed by gynecologic oncologists, pathologists, and radiotherapists on the basis of the postoperative treatment plan, and observation, radiotherapy, or concurrent radiotherapy was recommended. Furthermore, symptomatic treatments, such as nutrition and psychological support, in MDT consultation can alleviate the symptoms of the patients, reduce their discomfort, and improve their quality of life.All 392 patients were unvaccinated. Among them, 65.82% had a KPS score of ≥ 80 at admission, 359 had a first episode, including 284 in stages IA–IIA2, 75 in stages IIB–IVB, and 33 had a relapse, with a minimum of 2 days and a maximum of 2.5 months from diagnosis to treatment. We analyzed the clinical data of 416 MDT cases of cervical cancer and compared the outcomes of 364 cases with implemented decisions and the 52 cases without implemented decisions. We found that patients with implemented decisions were significantly better than those without implemented decisions in terms of diagnosis time, surgery efficiency, standardized radiotherapy, standardized chemotherapy, and other treatments. Moreover, the expected efficiency of full implementation was higher than that of partial implementation (93.99% vs. 72.84%), with a statistically significant difference . The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.JH completed most of the experiments, and XZ, HC, RL, and XW assisted in completing the experiments. YY, AC, DL, and CL completed data collection and statistics. JH and YP designed and supervised the completion of this experiment, and JH wrote this article. All authors contributed to the article and approved the submitted version.


Gastric cancer (GC) represents a malignancy with a multi-factorial combination of genetic, environmental, and microbial factors. Targeting lysosomes presents significant potential in the treatment of numerous diseases, while lysosome-related genetic markers for early GC detection have not yet been established, despite implementing this process by assembling artificial intelligence algorithms would greatly break through its value in translational medicine, particularly for immunotherapy.To this end, this study, by utilizing the transcriptomic as well as single cell data and integrating 20 mainstream machine-learning (ML) algorithms. We optimized an AI-based predictor for GC diagnosis. Then, the reliability of the model was initially confirmed by the results of enrichment analyses currently in use. And the immunological implications of the genes comprising the predictor was explored and response of GC patients were evaluated to immunotherapy and chemotherapy. Further, we performed systematic laboratory work to evaluate the build-up of the central genes, both at the expression stage and at the functional aspect, by which we could also demonstrate the reliability of the model to guide cancer immunotherapy.Eight lysosomal-related genes were selected for predictive model construction based on the inclusion of RMSE as a reference standard and RF algorithm for ranking, namely ADRB2, KCNE2, MYO7A, IFI30, LAMP3, TPP1, HPS4, and NEU4. Taking into account accuracy, precision, recall, and F1 measurements, a preliminary determination of our study was carried out by means of applying the extra tree and random forest algorithms, incorporating the ROC-AUC value as a consideration, the Extra Tree model seems to be the optimal option with the AUC value of 0.92. The superiority of diagnostic signature is also reflected in the analysis of immune features.In summary, this study is the first to integrate around 20 mainstream ML algorithms to construct an AI-based diagnostic predictor for gastric cancer based on lysosomal-related genes. This model will facilitate the accurate prediction of early gastric cancer incidence and the subsequent risk assessment or precise individualized immunotherapy, thus improving the survival prognosis of GC patients. Given the characteristics of gastric cancer itself, which is a malignant tumor caused by a combination of genetic, site-specific, and microbiological factors, the overall prognosis of gastric cancer patients has not improved significantly , 2. BothIn light of the emerging potential usefulness of lysosomes in the treatment of a wide range of malignancies and non-malignancies, such as neurological and cardiovascular diseases, targeting lysosomes presents a novel solution for the prevention of gastric cancer . Long thTherefore, we attempted to integrate and compare more than 20 mainstream classifying algorithms in machine learning to identify the most ideal diagnostic model for STAD, and subsequent insights involving the tumor immune microenvironment and drug sensitivity revealed the potential immunotherapeutic applicability of our lysosomal gene model. In addition, a series of validation of the model constructed genes for IFI30, including expression validation in different dimensions and functional assays, have more or less confirmed that the model constructed genes themselves serve as negligible risk factors for GC Figure 1https://www.gsea-msigdb.org/gsea/msigdb/cards/LYSOSOME) repository and the transcriptomic data with matching clinical information of Stomach Adenocarcinoma (STAD) from the TCGA (https://www.cancer.gov/tcga) cohort  approach to determine the optimal number of feature genes for model construction . Then, wAccording to the “No Free Lunch” theorem, we exhaustively ran out of 20 mainstream machine-learning algorithms to find the most ideal diagnostic model for STAD . The algA learning curve is a graphical depiction of the connection between proficiency and experience. Through machine learning, artificial intelligence mimicked human behavior, in essence. Therefore, by visualizing its learning process in the training set and its predictive performance in the validation set, we would be able to see if the model worked robustly in a direct manner.Usually, prognostic models and diagnostic tests are mathematically evaluated with measures of accuracy that do not consider clinical outcomes. To overcome this disadvantage, DCA, which is often used to compare the efficacy of different predictive models to maximize the clinical benefits when false positives and false negatives are inevitable, was introduced into the present study , 18.It is an extension of Gene Set Enrichment Analysis (GSEA) that produces distinct enrichment scores for every possible pairing of a sample and gene set , 20. EacTraditionally, the statistical principle of enrichment analysis is to use hypergeometric distribution to test the significance of a certain functional class in a group of genes. In recent years, scientists also tended to perform such analytics at a gene set level, treating numerous genes of interest as a whole. In the present study, the functional enrichment analysis was carried out not in both ways.The R package “ESTIMATE” was employed to calculate the scores of stromal cells, infiltrating immune cells, and tumor purity on the basis of gene expression. In this way, we unraveled the in-depth correlation between the LI and the surroundings of the malignant cells. The immune cell infiltration analysis was done by the CIBERSORT algorithm which is a widely used immunoinformatic tool to uncover the immunological implications of various diseases nowadays .Human gastric mucosal epithelial cells GES-1 and human gastric cancer cell lines SGC-7901, BGC-823, MGC-803, and HGC-27, which were identified by DNA typing of STR sequences, were purchased from Shanghai Institutes of Biological Sciences (CAS). The above cells were cultured at 37°C in a humidified incubator containing 5% CO2, cultured with DMEM (HyClone) mixed with 10% fetal bovine serum , where the culture medium was changed once a day.After extraction of total RNA using RNAiso Plus , RT-qPCR was performed using Revertaid First Strand cDNA synthesis kit , which was conducted under the manufacturer’s protocol. An SYBR Green-based real-time fluorescent quantitative polymerase chain reaction was carried out using GAPDH as an internal reference, in which the primers involved were GAPDH-F: GGTGAAGGTCGGTGTGAACG; GAPDH-R: ZCTCGCTCCTGGAAGATGGTG, IFI30-F: GTGGGAGTTCAAGTGCCAGCAT; IFI30-R: GCAGACAATGGTCAGGAAGGCT. The aforementioned results were calculated by the 2-ΔΔCT method to get the relative fold change of RNA expression.The target cells subjected to cold PBS wash were treated with cell lysis buffer according to the reagent manufacturer’s recommendation. And the extracted proteins were electrophoresed and separated on SDS-PAGE gels as background, and then transferred to PVDF membranes. After closed fixation with 5% bovine serum albumin, they were incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies.The five cells mentioned above GES-1, SGC-7901, BGC-823, MGC-803, and HGC-27, were treated as recommended by the reagent supplier and incubated with IFI30 primary antibody, and after 12 h incubation with specific secondary antibody at 37°C in the dark for 2 h. After staining with DAPI at room temperature, fluorescent images were captured with a confocal microscope system.Pre-fixed and paraffin-embedded pathological material was cut to 5 mm width and dewaxed. Endogenous peroxidase was inactivated with 3% H2O2. The treated slices were incubated overnight at 4°C with the corresponding protein antibodies, then incubated with secondary antibodies for 30 min followed by final chromogenic color development with freshly prepared DAB reagent.Transfected gastric cancer cells with high expression of IFI30 were inoculated in 6-well plates, and the cells were scratched manually to create scratches, which were placed under standard culture conditions for 48 h. Every 12 h, photographs were taken to observe the healing of the scratches. The concentration of 100,000 pretreated cells (100 μL) and the control cells were inoculated into transwell chambers, and the transwell assay and invasion assay were performed with or without matrix gel, after 24 h of cell growth, the cells were fixed with paraformaldehyde and stained with 0.05% crystalline violet for over 30 mins and then counted.The bioinformatics of this study involved was operated by R software as well as python software, t-test and Kruskal-wallis test were performed for the evaluation of pairwise transcriptomic data and Pearson or Spearman methods were employed for the evaluation of the correlation tests involved, p-values < 0.05 (*P < 0.05) were considered significant, for which **, p<0.01, ***, p < 0.001, and ****, p < 0.0001.Based on the stratification of tumor samples and healthy controls, we exhaustively screened the differentially expressed genes (DEGs), within which the majority of lysosome-related genes were presented  and signaling pathways were distinguished between the high- and low-LI groups. The results according to the GO database indicated that cornification was the most distinguishable biological process, followed by digestion, keratinocyte differentiation, muscle contraction, and keratinization  enables excellent immune assessment of patients as well as drug prediction, which would render a unique benefit to clinically intelligent adjunctive therapeutic approaches for GC patients, thereby facilitating the application of personalized management regimens.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Data retrieved from the TCGA database was collected from patients who provided informed consent based on guidelines laid out by the TCGA Ethics, Law, and Policy Group. The procedures used in this study adhere to the tenets of the Declaration of Helsinki and approval was obtained from the ethics committee of the Affiliated Hospital of Jiangsu University.QW, YL, ZZL, and YT designed the present study. QW, YL, ZZL, and YT prepared the figure and drafted this manuscript. XH, SZ, LW, and YL provided professional guidance in pathology. XH, SZ, LW, and YL are responsible for data curation and preprocessing. WL, HX, and ZZL are in charge for visualization. WL, HX, and ZZL enhanced the figures. WL, HX, and ZZL polished the language. ZRL and MX edited and revised the manuscript. ZRL and MX supervised the project. MX is responsible for funding acquisition. All authors contributed to the article and approved the submitted version.


Staphylococcus aureus (MRSA) is considered as a high priority by the World Health Organization. Furthermore, patients with a positive result for COVID-19 received early antibiotic treatment, a fact that potentially encourages the increase in antibiotic resistance. Therefore, there is an urgency to develop new drugs with molecular mechanisms different from those of the actual treatments. In this context, enzymes from the shikimate pathway, a route absent in humans, such as dehydroquinate dehydratase (DHQD), are considered good targets. In this work, a computer-aided drug design strategy, which involved exhaustive virtual screening and molecular dynamics simulations with MM-PBSA analysis, as well as an in silico ADMETox characterization, was performed to find potential noncovalent inhibitors of DHQD from MRSA (SaDHQD). After filtering the 997 million compounds from the ZINC database, 6700 compounds were submitted to an exhaustive virtual screening protocol. From these data, four molecules were selected and characterized (ZINC000005753647 (1), ZINC000001720488 (2), ZINC000082049768 (3), and ZINC000644149506 (4)). The results indicate that the four potential inhibitors interacted with residues important for substrate binding and catalysis, with an estimated binding free energy like that of the enzyme’s substrate. Their ADMETox-predicted properties suggest that all of them support the structural characteristics to be considered good candidates. Therefore, the four compounds reported here are excellent option to be considered for future in vitro studies to design new SaDHQD noncovalent inhibitors and contribute to the search for new drugs against MRSA.Bacteria resistance to antibiotics is a concerning global health problem; in this context, methicillin-resistant  Bacterial resistance to antibiotics is a concerning global health problem  that is Staphylococcus aureus, a pathogen considered a high priority by the World Health Organization [S. aureus (MRSA) strains, because this pathogen is resistant to most of the actual treatments, which in turn results in high rates of mortality [Even though effective antibiotics are a cornerstone of modern medicine, few bacteria can be as threatening as nization ,12 and iortality . Therefoortality . This paortality ,16.Salmonella (typhi and enterica), Escherichia coli, Clostridium difficile, and Staphylococcus aureus [One of these enzymes is 3-dehydroquinate dehydratase, which exists in two isoforms denoted as Type I and Type II. Structurally, Type I enzymes are homodimers, whilst Type II are dodecamers ,18. Types aureus ,20,21. Ds aureus ,23.S. typhi, S. flexneri, and S. auerus have suggested that this enzyme may act as a virulence factor [S. aureus, a Small Colony Variant (SCV) phenotype was isolated. This strain supports an Ochre mutation in the aroD gene, being auxotrophic for all aromatic amino acids and less virulent than the wild type [Francisella tularensis, the causative agent of Tularemia [In addition to the biosynthetic role of DHQD, deletions in the aroD gene in e factor . In the ild type . Furtherularemia . Therefoularemia . Staphylococcus aureus (SaDHQD). The molecules reported here potentially bind to the active site of the enzyme and support the structural characteristics to be considered promising drug candidates to fight against antibacterial resistance.In this work, a computer-aided drug design strategy, including exhaustive virtual screening, molecular dynamics, MM/PBSA analysis, and in silico ADMETox predictions, was followed to find potential noncovalent inhibitors of DHQD from methicillin-resistant C. difficile and E. faecalis [p = −1.7), therefore, compounds with a MW in the range of 200–250 Da, and a Log p value of −1 were included. Additionally, the highest reactivity clean criteria (without reactive groups) were applied, keeping only 123,000 molecules. Thereafter, Lipinski’s rule of five compliance, toxicity risk assessment, Topological Surface Area (TPSA), and rotatable bond filters were applied, leaving a final set of 6700 compounds. Nowadays, a lot of works using different virtual screening protocols to filter large chemical libraries have been published, but the challenge is always to select the best candidates for further experimental studies [ZINC000005753647 (1), ZINC000001720488 (2), ZINC000082049768 (3), and ZINC000644149506 (4)  inhibitors of SaDHQD have been reported . Noncovafaecalis ,29. With studies . In this studies . Here, w studies . The dat9506 (4) . At this moment, we have a set of possible compounds that might bind to SaDHQD. However, those ligands were selected using molecular docking that does not include sidechain and/or protein conformational changes that may influence the ligand’s binding to the enzyme . To gain3 and 4. However, the variations were so small that it can be said that all the ligands kept their binding sites. In fact, when ligand movement is observed in the binding site, complexes corresponding to substrate and compounds 3 and 4 show more variations  plots, which are used to explore the flexibility of each residue during simulations . TherefoThis is in consonance with data obtained from Radius of Gyration (RoG) plots. RoG is a quantity that can be related to the natural protein “breathing”. A high value of RoG might be an indication of a possible denaturation of the protein; the opposite might be related to a compaction of the protein. Initial RoG was 1.72 ± 0.003 nm. RoG values over time show fluctuations with increments above 1.8 nm. These fluctuations might indicate that the protein had an opening that can be related to the natural movement of the amino or carboxyl terminal. However, RoG distributions show that (even in the worst case: 3-dehydroquinate simulation in green), these values are located around 1.76 and 1.77 nm. These values show an increment of 0.4 Å when compared to the initial value. Therefore, no effect was observed in the compactness of the protein structure, i.e., the binding of the compounds did not generate a crucial conformational change .In respect to hydrogen bond formation, as was expected, none of the compounds made as many hydrogen bonds as the substrate . This is1 made a hydrogen bond with Glu35, whilst compound 2 did not make any interaction. Compound 3 made hydrogen bonds with Pro223 and Gln225, and compound 4 formed the same type of interaction with Glu35, Arg37, Arg70, His133, Lys160, and Gln225. Some of these interactions were shared by the substrate, such as Glu35, Lys160, and Gln225, additionally, a hydrogen bond with Arg37 and Arg202 was established by the substrate  and Brenk alerts that analyze the presence of undesirable substructures; synthetic accessibility, a characteristic that becomes important if the molecules reach the lead optimization step; Blood Brain Barrier penetration (BBB); and CYP450 inhibition, among others. From the fifteen characteristics evaluated, none of the compounds received a positive score in all of them; however, the four potential inhibitors approved the majority, highlighting PAINS alerts, synthetic accessibility, BBB, and Pgp inhibition  [p value of −1, considering 3-dehydroquinate properties (MW = 198 Da and Log p = −1.7), as well as structures without reactive groups (highest reactivity clean), were selected; this process resulted in a database of 123,000 compounds. Thereafter, additional filtering criteria, including physicochemical characteristics in accordance with the Lipinski rule of five, compounds with predicted toxicological parameters  were eliminated, structural aspects , and an adequate “druglike” value (−2 to 5) that indicates the potential that a certain compound had in order to become a drug, were applied using Data Warrior V.5.2.1 software (http://www.openmolecules.org/datawarrior/) [Compound selection was performed from the TRANCHES 3D collection in the ZINC15 database (er 2020) , which carrior/) . Finally3. To validate docking results, a crystallographic ligand was redocked, obtaining an RMSD value of 2.35 ± 0.00 Å. To consider a correct molecular docking study, the RMSD between the crystallographic and the redocked ligand must be lower than 2 Å. Our study had a higher RMSD value, but it is important to point out that we are not doing covalent docking, which is the case for 3-dehydroquinate. Nonetheless, the number and type of interactions found on the crystal and the redocked ligand remain at 82%. Only three interactions were missing on the redocked 3-dehydroquinate . Three independent docking experiments were made on each ligand (6700 × 3), and all outputs were clustered by binding score. Ligands with a binding score lower than the average minus two standard deviations  were selected for molecular dynamics simulations  was used as a receptor . The strulations .Complexes generated on Autodock Vina for each compound and the crystallographic ligand 3-dehydroquinate were used to perform molecular dynamics simulations using GROMACS 2019 ,51. To tAdditionally, binding free interaction energies of 300 random structures (100 structures of each replica) of the most populated cluster (for every replica) were calculated using the Molecular Mechanics/Poisson-Boltzmann Accessible Area (MM-PBSA) approximation ,37,54. MADMETox properties prediction  of the selected compounds was carried out using the SwissADME  and PREAZINC000005753647 (1), ZINC000001720488 (2), ZINC000082049768 (3), and ZINC000644149506 (4)). Clustering molecular dynamics show that these molecules were able, potentially, to interact with residues important for substrate binding and catalysis when compared to the crystallographic ligand. The interaction energy calculation for each residue in the binding site helped to detect hotspots for potential enzyme inhibition. Finally, the ADMETox properties prediction suggests that these compounds can be considered good candidates. Therefore, the four compounds reported here are excellent options to be considered for future in vitro studies to design new SaDHQD noncovalent inhibitors and contribute to the search for new drugs against MRSA.In the present work, a computer-aided drug design strategy was applied to report potential noncovalent SaDHQD inhibitors. After an exhaustive virtual screening protocol, four compounds were selected and characterized (


Spinal osteochondroma is rare. The purpose of this study is to examine the clinical characteristics and surgical treatment outcomes of 11 patients with spinal osteochondroma.The study included 11 patients with spinal osteochondroma. In these patients, we examined the onset level, onset site, initial symptoms, surgical procedure, outcomes and complications.Of the 11 patients, 9 presented with solitary tumours, and 2 had multiple. The mean post-operative observation period was six years and two months. The onset level was the cervical spine in eight patients, thoracic in two, and lumbar in one. The most common onset site was the posterior elements. The initial presentation was myelopathy in seven patients, radiculopathy in two, neck pain in one and feeling of mass in one. All patients underwent excision of the tumour, and depending on the tumour onset site, additional posterior or anterior decompression with or without fusion was performed. There was no recurrence in all patients. Intra-operative complications included dura tear and oesophageal injury in one patient with cervical onset, while post-operative complications included C5 palsy in one patient.In this study, surgical excision for osteochondroma of the spine were excellent with no recurrence of the tumour. These bone tumours generally arise in the metaphysis of long bones, presenting mature bone tissue that is continuous with the bone marrow and a cartilage cap. Spinal onset is rare, and less than 5% of solitary osteochondromas develop in the spine5.Osteochondroma (exostosis) is known to be the most common benign bone tumour, accounting for 10–15% of all bone tumours and 20–50% of all benign bone tumoursIn the present study, we examined the clinical characteristics and treatment outcomes of 11 patients with spinal osteochondroma who underwent surgical treatment and report our findings.6, and low back pain7  (JOA score), improvement according to the Hirabayashi method8, outcomes, and complications.The study included 11 patients with spinal osteochondroma (5 men and 6 women) who underwent surgical treatment from 1997 to January 2017. Of the 11 patients, 9 presented with solitary tumours, and 2 had multiple tumours. The mean age at the time of surgery was 43.4 (13–63) years, and the mean post-operative observation period was 3 years and 9 months, and the median was 6 years and 2 months (range: 1 year and 3 months to 12 years and 3 months). In these patients, we examined the onset level, onset site, initial symptoms, surgical procedure, Japanese Orthopaedic Association (JOA) score for cervical myelopathyThis study was approved by our institutional review board and written informed consent was obtained from all patients for the publication of this paper and any accompanying images.The onset level was the cervical spine in 8 patients, thoracic spine in two patients, and lumbar spine in one patient. The onset site was the lamina in five patients, vertebral body in one patient, facet joint in one patient, vertebral body and lamina in one patient, lamina and pedicle in two patients, and pedicle and head of the rib in one patient. The initial presentation was myelopathy in seven patients , radiculopathy in two patients , neck pain , and feeling of mass .For the surgical procedure, all patients underwent excision of the tumour, and depending on the tumour onset site, additional posterior decompression was performed in five patients, posterior decompression with fusion in three patients, and anterior fusion with posterior decompression and fusion in three patients.JOA scores changed from a mean of 14.4 points pre-operatively to 15.9 points post-operatively for the cervical spine, with a mean improvement rate of 57.7%; 6.5 points pre-operatively to 9.5 points post-operatively for the thoracic spine, with a mean improvement rate of 66.7%; and 23 points pre-operatively to 27 points post-operatively for the lumbar spine, with an improvement rate of 66.7% .In all patients, there was no recurrence observed during the follow-up observation period. Intra-operative complications in patients with cervical spine onset included dura tear and oesophageal injury in 1 patient, while post-operative complications included C5 palsy in one patient.For the case presentation, Case 9: the case subject was a 44-year-old woman with no medical history. She noticed numbness in the lower extremities at 32 weeks of pregnancy and developed buckling of the knees when walking at 33 weeks of pregnancy. Therefore, at 34 weeks of pregnancy, she consulted our department. Reduced thermal nociception below the level of the 9th thoracic vertebra and spastic partial paralysis of the bilateral lower extremities were observed. Magnetic resonance imaging (MRI) revealed a 11 × 9 × 26mm mass protruding into the spinal canal on the abdominal side of the lamina of the 9th/10th vertebra. The mass margin presented low signal intensity on both T1- and T2-weighted imaging, while part of the inner portion of the mass showed high signal intensity. The spinal cord was compressed, and intramedullary signal intensity change was observed on T2-weighted imaging . Plain cCase 4: The case subject was a 63-year-old woman. She consulted our department after noticing numbness on the left hand and weakness of the right lower extremity. She complained of numbness in the region of the C6 nerve, and impaired manual dexterity with mild gait impairment was observed. The tendon reflex was accentuated distal to the triceps reflex, and a pathological reflex was also observed. On MRI, a 9 × 4 × 8mm tumour protruding into the spinal canal was observed on the abdominal side of the lamina of the C5 vertebra. The tumour showed homogeneous low signal intensity on both T1- and T2-weighted imaging. The spinal cord was compressed, and on T2-weighted imaging, intramedullary signal intensity change was observed . CT afte10. Osteochondroma arising from the spine is relatively rare, accounting for 1–4% of solitary osteochondromas and 1–9% of HME10. It has been reported that the most common onset site is the cervical spine (~50% of cases), particularly at the C2 vertebra, followed by the thoracic and lumbar spine9. Furthermore, it is said that solitary osteochondroma commonly affects the cervical spine, whereas HME commonly affects the thoracic and lumbar spine10. Our patients also presented similar results. In the thoracic spine, the Th8 vertebra is most commonly affected, followed by the Th4 vertebra2. Among our patients with onset of the thoracic spine, onset was at the lower levels of the thoracic spine affecting the Th9 and Th10 vertebra. The reason that onset commonly affects the cervical spine is said to be attributed to great mobility, which places minute external force and stress on the epiphyseal cartilage, which leads to its dislocation11.Osteochondroma (exostosis) is the most common benign tumour, presenting as solitary tumours (90% of cases) and multiple tumours (10% of cases) in the form of hereditary multiple exostosis (HME)11. The fact that there is rare onset of osteochondroma in vertebral bodies can probably be explained by the absence of an epiphyseal plate2. In contrast, the reason that onset is common in the posterior elements of the spine is attributed to the fact that there are many secondary ossification centres  and that osteochondromas develop from the excessive cartilaginous tissue of such secondary ossification centres11. Furthermore, tumours protruding dorsally from posterior elements  are typically large and found as tumours that can be palpated, have superficial deformation in young individuals, and lack neurological symptoms1. In our patients, onset in the posterior elements, such as the lamina and facet joint, accounted to be about 80%. Osteochondroma progressing into the spinal canal, even if small, can cause concurrent neurological symptoms. Spinal osteochondroma causing myelopathy is considered rare9, with reports indicating an incidence of 0.5–1% in solitary lesions and approximately twofold in multiple lesions2, whereas a somewhat high incidence of 27.2–34% has also been reported in solitary spinal osteochondroma3. Of our 11 patients, 7 (63.6%) presented with myelopathy. This is thought to be attributed to persistent spinal cord compression when the tumour continues to grow even after completion of bone growth in spinal osteochondroma3. Furthermore, in our patients, the age of patients who developed myelopathy was older, at 35–63 years, with onset also found in elderly individuals, and it is possible that age-related degenerative changes in the spine contributed to myelopathy onset12.With regard to onset site, tumours most commonly develop in the posterior elements and protrude dorsally, whereas anterior onset, such as of the vertebral body, is rare5, which were similar to the findings in our patients. MRI findings include low signal intensity on T1-weighted imaging, reflecting a cartilage cap on the osteochondroma with cartilage rich in water content, and high signal intensity on T2-weighted imaging, while the marrow part presents high signal intensity on both T1- and T2-weighted imaging due to rich fatty content13. Furthermore, it is important to evaluate the thickness of the cartilage cap, and lesions with a cartilage cap exceeding 1 or 3cm have an increased risk of malignant transformation14.In the diagnosis of spinal osteochondroma, CT is particularly useful. Typical CT findings include the following: (1) round tumour with distinct border, (2) bone-like concentration with diffuse calcification, (3) paravertebral, dumbbell type, or eccentric intraspinal onset, and (4) continuity of the lesion with the cortical bone and medullary space3. For osteochondroma, it is important that the cartilage cap is completely surgically removed to prevent recurrence14. Tumours generally develop in the posterior elements; therefore, full or partial laminectomy, depending on the size of the lesion and removal of the spinous process by posterior approach is often used for marginal resection; however, depending on the onset site of the tumour, an anterior or anteroposterior approach will be needed3. Furthermore, when the facet joint is resected, spinal fusion will be required14.Surgery is indicated for tumours developing within the spinal canal; those compressing the spinal cord, cauda equina, and nerve root with neurological symptoms; those suspected of malignant transformation; and those with pain and restricting range of motion11, and the mean duration until recurrence ranges from 1 to 26 years2. Furthermore, the incidence of malignant transformation is 1–3% in solitary osteochondroma3 and 10–12% in HME11. In our patients, during the mean post-operative follow-up observation period of six years and two months, there was no recurrence observed; however, we believe that, in the future, long-term follow-up observation is needed.The rate of recurrence following osteochondroma excision is 1.3–8%Osteochondroma is relatively common bone tumour, accounting for 20-50% of all benign bone tumours, but occurs infrequently in the spine accounting for less than 5% of solitary osteochondromas. After examining the clinical characteristics and treatment outcomes of 11 patients with spinal osteochondroma who underwent surgical treatment, we found that these lesions caused neurologic symptoms including myelopathy (63.6%) and radiculopathy (18.2%). All patients underwent tumour excision including cartilaginous cap and no recurrence was observed. When tumour excision is performed adequately, the outcomes are excellent with very low recurrence rates.


Kaempferol, a secondary metabolite found in plants, is a naturally occurring flavonoid displaying significant potential in various biological activities. The chemical structure of kaempferol is distinguished by the presence of phenyl rings and four hydroxyl substituents, which make it an exceptional radical scavenger. Most recently, an increasing number of studies have demonstrated the significance of kaempferol in the regulation of intestinal function and the mitigation of intestinal inflammation. The focus of the review will primarily be on its impact in terms of antioxidant properties, inflammation, maintenance of intestinal barrier function, and its potential in the treatment of colorectal cancer and obesity. Future research endeavors should additionally give priority to investigating the specific dosage and duration of kaempferol administration for different pathological conditions, while simultaneously conducting deeper investigations into the comprehensible mechanisms of action related to the regulation of aryl hydrocarbon receptor (AhR). This review intends to present novel evidence supporting the utilization of kaempferol in the regulation of gut health and the management of associated diseases. The gastrointestinal tract represents the most extensive and delicate interface with the external environment . This inIt has been reported that nutritional or natural antioxidant medical interventions have demonstrated efficacy in enhancing intestinal health and the prevention or treatment of intestinal diseases ,11. FlavH-chromen-4-one), also known as indigo yellow, is a flavonoid compound with a molecular structural formula of C15H10O6 and a relative molecular weight of 286.23. Its pure product form is a yellow crystalline powder with a melting point of 276–278 °C and an acidity coefficient of 6.34 ± 0.40 [Kaempferol -44 ± 0.40 ,17. It i4 ± 0.40 .Kaempferia galanga [Kaempferol initially originated from the rhizomes of  galanga . Further galanga ,16. The  galanga . Additio galanga . Black t galanga . Spices  galanga . The extCichorium) resulted in a urinary excretion of 1.9% within 24 h when taking 9 mg of kaempferol. Furthermore, the peak concentration in plasma reached 0.1 μM after 5.8 h [Kaempferol possesses a relatively high affinity for lipids, and as a result, it is predominantly absorbed in the small intestine via active transport, passive diffusion, and facilitated diffusion ,20. Accoer 5.8 h .Research conducted on the metabolism and excretion of kaempferol has indicated that it undergoes primary metabolism within the liver and subsequently circulates in the bloodstream in the form of methyl, sulfate, or glucuronide conjugates ,25. In tThere has been substantial research conducted on kaempferol due to its various bioactivities, such as its antioxidant, antimicrobial, anti-inflammatory, and anticancer properties . KaempfeOxidative stress is a state of imbalance between cellular oxidants and antioxidants, leading to inadequate elimination of reactive oxygen (ROS) and nitrogen species (RNS) produced within the cells . The exc2+ levels while increasing cell viability [GCLC mRNA levels while simultaneously increasing T-Nrf2 protein levels as well as GSR, GSTA4, and HO-1 mRNA levels [Cyclin D1, CDK4, CDK6 mRNA levels, and β-catenin protein level in the cells [In a study, it was revealed that treating IEC-6 cells  with 5 μmol/L of kaempferol for 24 h resulted in a significant reduction in oxidative damage and cell cytotoxicity when challenged with indomethacin . More spiability . Similariability . The treA levels . FurtherA levels . Additiohe cells .Based on the aforementioned results, kaempferol has the potential to augment the antioxidant capacity and reduce oxidative stress in intestinal epithelial cells. These positive effects can be mainly attributed to its activation of the Nrf2 pathway, which is considered the primary cellular endogenous pathway for antioxidants . Once thInflammation is an essential biological response that is accompanied by symptoms including pain, redness, and heat. This response frequently leads to the typical physiological impairment of the affected tissues , such asIL-1β, IL-6, TNF-α, COX-2, MCP-1, iNOS, TLR4, NLRP3, MAPK, and NF-κB, while there was an increase in the mRNA level of IL-10 in the colonic tissues of mice with colitis induced by DSS [One approach for drug-giving is oral administration, and the ingestion of kaempferol as a means of oral administration has been documented as a viable technique for alleviating intestinal inflammation. In a recent study, it was observed that the oral administration of kaempferol at a dose of 100 mg/kg effectively inhibited acute colitis and extended the length of the colon in mice treated with dextran sulfate sodium (DSS). The authors provided evidence supporting the proposition that the positive impacts of kaempferol on colitis are mediated via the regulation of the efflux transporters BCRP (breast cancer resistance protein) and MRP2 (multi-drug resistance associated protein 2) . Most red by DSS . Furtherd by DSS . Signifid by DSS . More ind by DSS . Additiod by DSS .2 (prostaglandin E2) and LTB4 (leukotriene B4), as well as a decrease in colonic mucosa MPO level. The study also found that the mRNA levels of COX-2, iNOS, TNF-α, IL-1β, and IL-6 were downregulated, whereas the mRNA level of TFF3 was upregulated in the colonic mucosa of mice receiving the 0.3% kaempferol supplementation [F4/80, IL-6, TNF-α, MCP-1, and IL-1β mRNA levels in the colon tissue of the mice [The dietary supplementation of kaempferol has also been found to ameliorate intestinal inflammation induced by DSS and obesity in a mouse model ,39. It wentation . Regardientation . In an eIL-6 and RELA/p65. The treatment also led to an increase in the levels of IL-10 in the culture supernatant [The anti-inflammatory properties of kaempferol on the intestine were further demonstrated in in vitro investigations. According to a recent study, the administration of 50 μmol/L kaempferol for a duration of 3 h significantly reduced the production of cell cytokines  and adhesion proteins  in rat intestinal microvascular endothelial cells that were subjected to challenge by LPS (lipopolysaccharide) . As for ernatant . More inernatant .Based on the in vivo and in vitro studies mentioned above, it is evident that kaempferol possesses the potential to alleviate intestinal inflammation across various inflammation models, such as LPS, TNF-α, DSS, and high-fat diet-induced inflammation. Nevertheless, further clinical studies are imperative to endorse the implementation of kaempferol as a preventive or therapeutic agent for intestinal ailments associated with inflammation.The intestine serves as the primary organ responsible for the absorption of nutrients and water, making it the most extensive region in contact with environmental elements . The intIt was discovered that the oral administration of doses of 25, 50, or 100 mg/kg/day effectively reduced the levels of serum D-lactate and FITC-dextran in C57BL/6J mice with chronic colitis induced by DSS . This deE. coli in IEC-6 cells [In vitro studies further confirm the advantageous impact of kaempferol on the intestinal barrier function. Suzuki and colleagues (2011) conducted a study in which they observed that the treatment of Caco-2 cells with 100 μmol/L kaempferol for a duration of 48 h resulted in an improvement in intestinal barrier function . The resThe impairment of the intestinal barrier function has been documented in diquat , indometIn this context, kaempferol has the ability to preserve the integrity of the intestinal barrier both in vivo and in vitro. Additionally, it possesses the ability to alleviate disruptions in the intestinal barrier that are induced by various stimuli, such as diquat, indomethacin, LPS, TNF-α, and deoxynivalenol. Nonetheless, the ongoing studies are presently constrained solely to mouse models and in vitro cell models, thus necessitating further human trials to authenticate the impact of introducing kaempferol.Min/+ miceApc [TNF-α in the ileum of the mice. Moreover, it was noted that the levels of FXR mRNA and protein were upregulated, while the level of β-Catenin protein in the intestinal tissues was downregulated due to kaempferol administration [According to multiple research sources, it has been proposed that kaempferol demonstrates promising qualities as an anticancer agent . It was n/+ mice . The admstration .p53 gene (human colon cancer cells) [Multiple studies have been conducted at the cellular level regarding kaempferol’s impact on colorectal cancer. According to Cho and Park’s research conducted in 2013, it was observed that the administration of kaempferol led to a decline in cell proliferation and an increase in cell cycle arrest in HT-29 cells with the mutant r cells) . Moreover cells) . The autr cells) . The resr cells) . These fr cells) . Consistr cells) . Additior cells) .In light of the existing evidence, it is believed that kaempferol holds significant promise as a novel agent for the treatment of colorectal cancer. However, further extensive research is required to investigate the clinical trials and underlying mechanisms of action.The link between intestinal microbiota and metabolic disorder diseases, particularly in the case of obesity, has been evidenced in recent years . The datBacteroidetes and Proteobacteria, as well as a reduction in the relative abundance of Firmicutes at the phylum level in the collected fecal samples. Moreover, at the genus level, supplementation with kaempferol led to an increase in the relative abundance of Akkermansia, Bacteroides, and Lactobacillus in the fecal samples of mice that were fed a high-fat diet [Firmicutes and the decline in Bacteroidetes caused by the high-fat diet. The ratio of Firmicutes to Bacteroidetes (F/B) escalated with the consumption of the high-fat diet. Conversely, the supplementation of kaempferol diminished the F/B ratio to a degree comparable to the level seen in mice from the control group. Kaempferol also effectively counteracted the alterations induced by a high-fat diet in Rikenallaceae, Prevotellaceae, Desulfovibrionaceae, and Helicobacteraceae at the family level. At the genus level, the inclusion of kaempferol significantly mitigated the alterations observed in Alistipes, Lachnospiraceae_NK4A136_group, Romboutsia, and Faecalibaculum. Moreover, kaempferol substantially reversed the modifications induced by a high-fat diet in Desulfovibrio, Helicobacter, and Akkermansia. The authors employed Spearman’s correlation analysis to demonstrate that the prevalence of various bacterial strains had a significant correlation with physiological indicators linked to obesity. Specifically, Parasutterella, Alloprevotella, and Akkermansia displayed a negative correlation with obesity-associated indices. These findings suggest that these particular strains might exert a crucial influence in mitigating obesity [Kaempferol is involved in the regulation of obesity via the regulation of the microbiota. In a study, it was discovered that the addition of kaempferol at a dosage of 200 mg/kg in the diet for a duration of 8 weeks led to a reduction in body weight, as well as the weights of inguinal white adipose tissue, epididymal white adipose tissue, and perirenal white adipose tissue in C57BL/6J mice that were subjected to a high-fat diet . The autfat diet . In anotfat diet . The inv obesity .Hence, kaempferol participates in the regulation of obesity via the regulation of gut microbiota. The anti-obesity property of kaempferol also suggests that the gut microbiota could be regarded as a potential target for tackling obesity, with the aim of investigating potential agents, particularly plant-derived polyphenols, to combat obesity.VEGFR-2, VEGF mRNA, VEGFR-2, p-Akt/Akt, VEGF-A, HIF-1α (hypoxia-inducible factor 1α), and p-p38/p38 protein in the cells [Additional physiological functions of kaempferol in the intestine have been documented as well. It was reported that oral administration of 25, 50, or 100 mg/kg/day demonstrated a mitigating effect on intestinal angiogenesis in mice. This was evidenced by a reduction in VEGF-A  and NO levels, an increase in IL-22BP (interleukin-22 binding protein) protein level, and a decrease in VEGFR2 , COX-2 (cyclooxygenase-2), TRAF6 (TNF receptor-associated factor 6) protein levels, as well as VEGFR2 (IHC score) and microvascular density of the mice . In a rehe cells . Furtherhe cells . Additiohe cells . In anothe cells . The admhe cells .These recent functional findings expand the range of potential applications of kaempferol while enhancing our comprehension of this compound. They provide compelling evidence for the need to conduct further comprehensive research on kaempferol’s role in regulating intestinal function. The condensation of kaempferol’s effectiveness as a therapeutic agent for intestinal health and diseases demonstrated in both in vivo and in vitro studies, has been compiled and presented in MinApc/+ mice is achieved via the modulation of gut microbiota and bile acid signaling [Regarding the further investigation of the mechanism of action, more attention should be given to the modulatory effects of kaempferol on gut microbiota and microbiota-derived metabolites. The role of gut microbiota in intestinal inflammation and intestinal barrier function, and consequently the overall health of the host body, has been firmly established ,60,61. Iignaling . Furtherignaling .Most recently, the targeting of the aryl hydrocarbon receptor (AhR) with gut microbiota phenolic metabolites has been recognized as a crucial mechanism in the prevention and treatment of intestinal diseases ,63. CurrAdditionally, despite the thorough validation of the efficacy of kaempferol in regulating intestinal physiological function and diseases via in vitro cell experiments and in vivo murine model experiments, no data are available regarding in vivo human trials. Furthermore, an extensive number of trials is required to investigate the precise dosage and duration of kaempferol in various pathological conditions. More importantly, there is a crucial need to examine the mechanism by which kaempferol regulates the specific pathologies of the intestine, with the aim of enhancing disease prevention and treatment, as well as obtaining a comprehensive understanding of specific intestinal diseases. Overall, future research endeavors should also prioritize investigating the specific dosage and duration of kaempferol in different pathological conditions, along with delving further into the underlying mechanisms of action that involve AhR regulation.Kaempferol is a naturally occurring flavanol that is widely distributed across various plant genera as a secondary metabolite. Its chemical structure is characterized by phenyl rings and four hydroxyl substituents, making it an excellent radical scavenger. Kaempferol signifies a natural and potent antioxidant that possesses potential as a suitable functional agent for intestinal health. Numerous researchers have substantiated the idea that kaempferol holds significant potential for augmenting antioxidant capacity, mitigating gut inflammation, enhancing barrier function, and even aiding in the treatment of colorectal cancer and obesity. Based on current discoveries, the development and application of kaempferol emerges as a promising and evolving avenue for future research on intestinal health. Upcoming research endeavors should also prioritize investigating the precise dosage and duration of kaempferol administration in varying pathological conditions, while simultaneously delving deeper into the comprehensible mechanisms of action that involve AhR regulation.