AIM To screen the material basis for Rhei Radix et Rhizoma on improving human umbilical vein endothelial cell(HUVECs)injury before and after carbonization.METHODS The HPLC fingerprints were established,after which difference analysis was performed by orthogonal partial least squares discriminant analysis.The LPS-induced HUVECs injury model was established,then NO,MDA levels and SOD activity were detected.Gray correlation analysis and partial least squares regression were adopted in the investigation of spectrum-effect relationship,and active constituents were screened.RESULTS There were 22 and 20 common peaks in the fingerprints for 17 batches of raw products and carbonized products,respectively,along with the similarities of more than 0.8.Twelve main difference components were observable,among which gallic acid,5-hydroxymethylfurfural,catechin,aloe rhodopsin-8-O-β-D-glucoside,rhubarbic acid-8-O-β-D-glucoside and sennosides A were identified.The carbonized products demonstrated stronger effect on improving HUVECs injury than the raw product.The correlations of common peaks were more than 0.5 in the fingerprints for raw products and carbonized products,and peaks 3,5,11,12,15,19,23 exhibited significant effects on their efficacy(VIP values＞1).CONCLUSION This accurate,reliable and reproducible method can provide a basis for clarifying the material basis for hemostatic efficacy of Rhei Radix et Rhizoma and its carbonized product.

Objective:To analyze the risk factors for fat liquefaction after surgery for pediatric appendiceal abscess and to construct and validate a predictive model for post-operative fat liquefaction.Methods:The training set collected clinical data from 60 children who developed fat liquefaction after appendiceal abscess surgery and 60 children who did not develop fat liquefaction for comparison.The validation set collected data from 97 children during the same period ac-cording to the same criteria to test the model's efficacy.LASSO logistic regression was used to screen potential diag-nostic factors,and a Logistic regression model was employed for univariate analysis.Collinearity diagnostic tests were conducted among the risk factors.The stepwise regression method using the training dataset was applied to evaluate the importance of each risk factor for fat liquefaction.A multivariate Cox proportional hazards model was used to calcu-late the concordance index of the risk factors in both the training and validation sets.A predictive model was constructed using Logistic regression,and the clinical value of the predictive model was evaluated using ROC curves,calibration curves,and decision curves.Results:Compared with the non-liquefaction group,the liquefaction group had higher val-ues in terms of body weight,heart rate,diameter of the tender mass in the right lower abdomen,degree of appendiceal thickening,body temperature upon admission,fasting plasma glucose(FPG)level,surgical duration,intraoperative blood loss,wound healing time,length of hospital stay,volume of abdominal drainage fluid,and infection status(P<0.05).LASSO logistic regression analysis identified potential factors including body temperature upon admission,heart rate,body weight,diameter of the tender mass in the right lower abdomen,degree of appendiceal thickening,FPG level,sur-gical duration,and intraoperative blood loss.Univariate analysis revealed that body weight,diameter of the tender mass in the right lower abdomen,FPG level,surgical duration,and intraoperative blood loss were influencing factors for fat lique-faction after appendectomy(P<0.05).Body weight and FPG factors were mutually independent with no multicollinearity,while there was multicollinearity among the diameter of the tender mass in the right lower abdomen,surgical duration,and intraoperative blood loss.The risk model combining the diameter of the tender mass in the right lower abdomen,in-traoperative blood loss,FPG,surgical duration,and body weight had the highest concordance index in both the training and validation sets,with values of 0.811 and 0.814,respectively.The Logistic regression model established the predictive model as Logit(P)=-1.136+0.664×(diameter of the tender mass in the right lower abdomen)+0.449×(surgical duration)+0.622×(intraoperative blood loss)+0.200×(body weight)+0.578×(FPG).The area under the ROC curve was 0.920(95%CI:0.869～0.942),with a sensitivity of 93.14%and a specificity of 85.73%.The calibration curve of the predictive model showed good fit with the ideal curve.The predictive model had a high net benefit.Conclusion:Body weight,diameter of the tender mass in the right lower abdomen,FPG level,surgical duration,and intraoperative blood loss are risk factors for fat liquefaction after appendectomy and have good predictive value for the occurrence of fat liquefaction.

Objective:To analyze the risk factors for fat liquefaction after surgery for pediatric appendiceal abscess and to construct and validate a predictive model for post-operative fat liquefaction.Methods:The training set collected clinical data from 60 children who developed fat liquefaction after appendiceal abscess surgery and 60 children who did not develop fat liquefaction for comparison.The validation set collected data from 97 children during the same period ac-cording to the same criteria to test the model's efficacy.LASSO logistic regression was used to screen potential diag-nostic factors,and a Logistic regression model was employed for univariate analysis.Collinearity diagnostic tests were conducted among the risk factors.The stepwise regression method using the training dataset was applied to evaluate the importance of each risk factor for fat liquefaction.A multivariate Cox proportional hazards model was used to calcu-late the concordance index of the risk factors in both the training and validation sets.A predictive model was constructed using Logistic regression,and the clinical value of the predictive model was evaluated using ROC curves,calibration curves,and decision curves.Results:Compared with the non-liquefaction group,the liquefaction group had higher val-ues in terms of body weight,heart rate,diameter of the tender mass in the right lower abdomen,degree of appendiceal thickening,body temperature upon admission,fasting plasma glucose(FPG)level,surgical duration,intraoperative blood loss,wound healing time,length of hospital stay,volume of abdominal drainage fluid,and infection status(P<0.05).LASSO logistic regression analysis identified potential factors including body temperature upon admission,heart rate,body weight,diameter of the tender mass in the right lower abdomen,degree of appendiceal thickening,FPG level,sur-gical duration,and intraoperative blood loss.Univariate analysis revealed that body weight,diameter of the tender mass in the right lower abdomen,FPG level,surgical duration,and intraoperative blood loss were influencing factors for fat lique-faction after appendectomy(P<0.05).Body weight and FPG factors were mutually independent with no multicollinearity,while there was multicollinearity among the diameter of the tender mass in the right lower abdomen,surgical duration,and intraoperative blood loss.The risk model combining the diameter of the tender mass in the right lower abdomen,in-traoperative blood loss,FPG,surgical duration,and body weight had the highest concordance index in both the training and validation sets,with values of 0.811 and 0.814,respectively.The Logistic regression model established the predictive model as Logit(P)=-1.136+0.664×(diameter of the tender mass in the right lower abdomen)+0.449×(surgical duration)+0.622×(intraoperative blood loss)+0.200×(body weight)+0.578×(FPG).The area under the ROC curve was 0.920(95%CI:0.869～0.942),with a sensitivity of 93.14%and a specificity of 85.73%.The calibration curve of the predictive model showed good fit with the ideal curve.The predictive model had a high net benefit.Conclusion:Body weight,diameter of the tender mass in the right lower abdomen,FPG level,surgical duration,and intraoperative blood loss are risk factors for fat liquefaction after appendectomy and have good predictive value for the occurrence of fat liquefaction.

AIM To screen the material basis for Rhei Radix et Rhizoma on improving human umbilical vein endothelial cell(HUVECs)injury before and after carbonization.METHODS The HPLC fingerprints were established,after which difference analysis was performed by orthogonal partial least squares discriminant analysis.The LPS-induced HUVECs injury model was established,then NO,MDA levels and SOD activity were detected.Gray correlation analysis and partial least squares regression were adopted in the investigation of spectrum-effect relationship,and active constituents were screened.RESULTS There were 22 and 20 common peaks in the fingerprints for 17 batches of raw products and carbonized products,respectively,along with the similarities of more than 0.8.Twelve main difference components were observable,among which gallic acid,5-hydroxymethylfurfural,catechin,aloe rhodopsin-8-O-β-D-glucoside,rhubarbic acid-8-O-β-D-glucoside and sennosides A were identified.The carbonized products demonstrated stronger effect on improving HUVECs injury than the raw product.The correlations of common peaks were more than 0.5 in the fingerprints for raw products and carbonized products,and peaks 3,5,11,12,15,19,23 exhibited significant effects on their efficacy(VIP values＞1).CONCLUSION This accurate,reliable and reproducible method can provide a basis for clarifying the material basis for hemostatic efficacy of Rhei Radix et Rhizoma and its carbonized product.

Based on the strategy of metabolomics combined with bioinformatics, this study analyzed the potential allergens and mechanism of pseudo-allergic reactions (PARs) induced by the combined use of Reduning injection and penicillin G injection. All animal experiments and welfare are in accordance with the requirements of the First Affiliated Experimental Animal Ethics and Animal Welfare Committee of Henan University of Chinese Medicine (approval number: YFYDW2020002). Based on UPLC-Q-TOF/MS technology combined with UNIFI software, a total of 21 compounds were identified in Reduning and penicillin G mixed injection. Based on molecular docking technology, 10 potential allergens with strong binding activity to MrgprX2 agonist sites were further screened. Metabolomics analysis using UPLC-Q-TOF/MS technology revealed that 34 differential metabolites such as arachidonic acid, phosphatidylcholine, phosphatidylserine, prostaglandins, and leukotrienes were endogenous differential metabolites of PARs caused by combined use of Reduning injection and penicillin G injection. Through the analysis of the "potential allergen-target-endogenous differential metabolite" interaction network, the chlorogenic acids (such as chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and isochlorogenic acid A) and β-lactam allergens in the combination of the two may be mainly regulated by PLD1, PLA2G12A and CYP1A1. The three upstream signal target proteins mainly activate the arachidonic acid metabolic pathway, promote the degranulation of mast cells, release downstream endogenous inflammatory mediators, and induce PARs.

Traditional Chinese medicine(TCM) has been treating diseases for thousands of years and still holds an irreplaceable position in modern disease treatment. The composition of TCM is complex, and its mode of action features multi-pathway and multi-target characteristics, making its pharmacological actions and mechanisms complicated. The principles and mechanisms of TCM have always been a hot topic among scholars from various fields. Increasingly, studies show that indirect action plays an important role in the mode of action of TCM. The gut microbiota is regarded as a new invisible metabolic organ, and TCM can treat diseases by targeting the gut microbiota, thereby exerting an indirect effect across organs based on the connection between organs. Therefore, this paper summarized the indirect regulatory mechanisms of TCM based on the visceral manifestation theory and the organ axis of modern medical research. It also discussed the talks and communication mechanisms between the gut microbiota and target organs such as the liver, brain, kidney, and lungs, providing references for research on the mechanisms of indirect action of TCM and the development of innovative drugs.
Gastrointestinal Microbiome/drug effects* ; Humans ; Medicine, Chinese Traditional ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Kidney/metabolism* ; Liver/drug effects* ; Brain/drug effects* ; Lung/drug effects*

Objective:To investigate the knowledge of Sixth Chinese national consensus report on the management of Helicobacter pylori infection ( treatment excluded) (hereinafter referred to as sixth national consensus) and 2022 Chinese national clinical practice guideline on Helicobacter pylori eradication treatment (hereinafter referred to as the guideline)among medical staff from general hospitals in Hainan. Methods:From February 20 to May 7, 2023, a questionnaire survey on the diagnosis and treatment of Helicobacter pylori ( H. pylori) infection was conducted among 1 463 medical staff from 15 general hospitals in Hainan Province. The questionnaire was drawn up according to the sixth national consensus and the guideline, covering knowledge of 6 sections, induding H. pylori related diseases, detection of H. pylori, eradication, prevention and influence factors of eradication of H. pylori, etc. Chi-square test was used for statistical analysis. Results:A total of 1 463 valid questionnaires were collected with the effective responsive rate of 100.00%.The 1 463 subjects included 225 gastroenterologists and 1 238 other medical staff(including 503 physicians from other departments, 264 surgeons and 471 medical technologists and pharmacists). About 78.67%(177/225)of gastroenterologists agreed that the overall infection rate of H. pylori in China was more than 20%, the awareness rate was higher than that of other medical staff (physicians from other departments 65.41%(329/503), surgeons 61.74%(163/264), medical technologists and pharmacists 60.30%(284/471); the following datas were sorted by this position), and the difference was statistically significant ( χ2=30.97, P<0.001). About 51.11%(115/225) of gastroenterologists considered that H. pylori serological antibody test could not be used as a diagnostic method for current infection, the awareness rate was higher than that of other medical staff(22.07%(111/503), 14.02%(37/264), 12.31%(58/471)), and the difference was statistically significant( χ2 =152.66, P<0.001). Proton pump inhibitor and potassium-competitive acid blocker should be discontinued for 2 weeks, and antibiotics and bismuth should be discontinued for 4 weeks before urea breath test, and the awareness rates of gastroenterologists were higher than those of other medical staff (38.67%(87/225) vs. 23.26%(117/503), 19.70%(52/264), 18.47%(87/471); 60.89%(137/225) vs. 26.64%(134/503), 25.76%(68/264), 23.78%(112/471)), and the differences were statistically significant ( χ2 =133.70 and 165.51, both P<0.001). For refractory H. pylori infection, 98.67%(222/225)of gastroenterologists agreed with the individualized diagnosis and treatment of H. pylori infection should be guided by bacterial culture, antibiotic susceptibility test or drug resistance gene test, and the awareness rate was higher than that of other medical staff (91.85%(462/503), 93.56%(247/264), 93.21%(439/471)), and the difference was statistically significant( χ2=20.55, P=0.002). About 70.67% (159/225) of gastroenterologists recommended a bismuth containing quadruple regimen, 80.44% (181/225) supported a 10 to 14 day H. pylori eradication course, and the awareness rates were higher than other medical staff (46.92%(236/503), 33.33%(88/264), 32.91%(155/471); 67.20%(338/503), 59.09%(156/264), 53.93%(254/471)), and the differences were statistically significant ( χ2=111.25 and 59.99, both P<0.001). The understanding rates of the sixth national consensus and the guideline in gastroenterologists was 85.33% (192/225), which was higher than that of other medical staff (64.21%(323/503), 66.67%(176/264), 57.96%(273/471)), and the difference was statistically significant ( χ2=85.47, P<0.001). Conclusions:Gastroenterologists from general hospitals in Hainan Province have a better understanding of the sixth national consensus and the guideline than other medical staff. However, there is still a lack of deep understanding of the sixth national consensus and the guideline, and it is necessary to further strengthen the learning and application of the sixth national consensus and the guideline.

Objective To explore the mechanism of Zhuangxuan Yin in the treatment of children with H1N1 pneumonia through regulating gut microbiota mediated by p38 mitogen-activated protein kinase(p38-MAPK)pathway.Methods A BALB/c mouse model of H1N1 pneumonia was prepared using the H1N1 influenza virus allantoic solution for nasal drop.The model was randomly separated into 5 groups:model group,Zhuangxuan Yin low-,medium-and high-dose groups,and high-dose Zhuangxuan Yin(28.66 g·kg-1)+anisomycin(10 mg·kg-1)group.The control group was infused with sterile physiological saline of equal volume using the same method.After treatment with Zhuangxuan Yin and anisomycin,the lung index of mice in each group was measured,and HE staining was applied to detect the pathological morphology of lung and large intestine tissues.16SrRNA gene sequencing was applied to detect the structural difference of gut microbiota in mice of each group.Enzyme-linked immunosorbent assay(ELASA)was applied to measure the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-4,IL-6,and IL-1β in bronchoalveolar lavage fluid(BALF)and large intestine tissue of mice in each group.Western Blot was applied to detect the expression of p38-MAPK pathway-related proteins in lung and large intestine tissues of mice in each group.Results Compared with the control group,the lung and large intestine tissues of the model group mice showed obvious pathological damage,the lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues increased(P＜0.05).The abundance of class Bacteroidales decreased(P＜0.05).Compared with the model group,the pathological damage in the lung and large intestine tissues of mice in the Zhuangxuan Yin low-,medium-and high-dose groups were reduced.The lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues decreased(P＜0.05),the abundance of class Bacteroidales increased(P＜0.05),and in a dose-dependent manner(P＜0.05).Compared with the high-dose Zhuangxuan Yin group,the pathological damage in the lung and large intestine tissues of mice in the high-dose Zhuangxuan Yin+anisomycin group was aggravated,the lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues increased(P＜0.05),and the abundance of class Bacteroidales decreased(P＜0.05).Conclusion Zhuangxuan Yin can improve the imbalance of intestinal microbiota in H1N1 pneumonia mice by inhibiting p38-MAPK signal activation,thereby inhibiting inflammation and reducing lung and large intestine tissue damage in mice,which may have a therapeutic effect on children with H1N1 pneumonia.

In mass spectrometry-based proteomics experiments,achieving high-throughput and efficientproteolytic digestion is crucial to ensure optimal protein cleavage and enhance the depth of protein identi-fication (including the number of identified proteins and the coverage of protein amino acid sequences) .Trypsin is the most widely used protease in mass spectrometry-based proteomics due to its ability to spe-cifically cleave the carboxyl terminus of arginine and lysine.However,it was found that Trypsin has some missed enzymatic efficiency for the cleavage of lysine residues.Therefore,in actual proteomics sample preparation,a combination of Trypsin and LysC will be used to ensure adequate cleavage of lysine resi-dues.Our study revealed that the commonly employed LysC-Trypsin tandem cleavage method exerts an impact on the enzymatic cleavage of protein samples by Trypsin due to the subsequent cleavage of Trypsin by initially added LysC.Consequently,we adjusted the order of LysC and Trypsin tandem digestion,with Trypsin cleavage being performed first followed by the addition of LysC to target any missed lysine resi-dues.We comprehensively compared and analyzed three distinct sequential digestion methods,namely Trypsin-Trypsin (T-T),LysC-Trypsin (L-T),and Trypsin-LysC (T-L),in terms of their effects on pro-tein sample preparation quality.The results demonstrated that the Trypsin-LysC sequential digestion ap-proach not only minimizes missed protein lysine/arginine cleavage sites without increasing experimental costs,at the same time yielding peptides with a moderate amino acid sequence length.The use of Tryp-sin-LysC digestion enhances the adsorption and separation of peptide samples in RP-HPLC,as well as improves the depth of protein detection and amino acid sequence coverage during tandem mass spectrome-try analysis.This research work offers a novel technical solution and serves as a valuable reference for proteome sample preparation.

Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder characterized by chronic hyperglycemia, hyperlipidemia, and peripheral insulin resistance. Endoplasmic reticulum stress (ERS), a response to cellular stress, is activated across various tissues during the progression of T2DM, leading to disruptions in protein synthesis. Notably, epithelial and endocrine cells with hormone-secreting functions are particularly vulnerable to functional impairments induced by ERS. The gut-pancreas axis is essential for regulating metabolism and the progression of T2DM. Intestinal epithelial L cells, integral to the intestinal barrier, can secrete the glucagon-like peptide-1 (GLP-1). This hormone promotes insulin secretion from pancreatic β-cells and plays a critical role in glucose metabolism. Importantly, ERS plays a critical role in regulating glucolipid-induced dysfunction of gut-pancreas axis. For instance, ERS is involved in regulating the intestinal barrier and the secretion of GLP-1 as well as insulin. Therefore, ERS can be a potential target for T2DM treatment. In this paper, we review the regulatory roles of ERS in the gut-pancreas axis during the development of T2DM, and summarize the therapeutic drugs and strategies targeting ERS for T2DM treatment.