Based on the pharmacokinetics theory, this study investigated the pharmacokinetic characteristics of albiflorin, paeoniflorin, wogonoside, and wogonin in normal and febrile rats and summarized absorption and elimination rules of Dayuanyin in them to provide reference for further development and clinical application of Dayuanyin. Blood samples were taken from the fundus venous plexus of normal and model rats after intragastric administration of Dayuanyin at different time points. The concentration of each substance in blood was determined by ultra performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS) technique at different time points. DAS 2.0, a piece of pharmacokinetics software, was used to calculate the pharmacokinetic parameters of each component. The results show that the 4 components had good linear relationship in their respective ranges, and the results of methodological investigation met the requirements. The pharmacokinetic parameters of C_(max), T_(max), t_(1/2), AUC_(0-t), AUC_(0-∞), and MRT_(0-t) were calculated by the DAS 2.0 non-compartmental model. Compared with those in the normal group, C_(max) and AUC_(0-t) of the 4 components in the model group were significantly increased. There were significant differences in the pharmacokinetic characteristics between the normal and model groups, suggesting that the absorption and elimination of Dayuanyin may be affected by the changes of internal environment of the body in different physiological states.
Animals ; Rats ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Rats, Sprague-Dawley ; Fever/metabolism* ; Tandem Mass Spectrometry ; Chromatography, High Pressure Liquid ; Glucosides/pharmacokinetics* ; Monoterpenes

The efficacy of Xiangmei Pills on rats with ulcerative colitis(UC) was investigated by characterizing the spectrum of the active chemical components of Xiangmei Pills. Rapid identification and classification of the main chemical components were performed,and the therapeutic effects of Xiangmei Pills on the proteins and intestinal flora of UC rats were analyzed to explore the mechanism of its action in treating UC. Fifty SD rats were acclimatized to feeding for 3 d and randomly divided into blank group, model group,mesalazine group(0. 4 g·kg~(-1)), low-dose group of Xiangmei Pills(1. 89 g·kg~(-1)), and high-dose group of Xiangmei Pills(5. 67 g·kg~(-1)), with 10 rats in each group. 5% dextrose sodium sulfate(DSS) was given by gavage to induce the male SD rat model with UC,and the corresponding medicinal solution was given by gavage after 10 days, respectively. The therapeutic effect of Xiangmei Pills on rats with UC was evaluated according to body mass, disease activity index(DAI), and hematoxylin-eosin(HE) staining, and the histopathological changes in the colon were observed. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) technique was used to rapidly and accurately identify the main chemical constituents of Xiangmei Pills. Immunohistochemistry was used to detect the expression of aryl hydrocarbon receptor(AhR),interferon-γ(IFN-γ), mucin-2(MUC-2), and cytochrome P450 1A1(CYP1A1) in colon tissue. Interleukin-22(IL-22) expression in colon tissue was detected by immunofluorescence. The 16S r DNA high-throughput sequencing technique was used to study the modulatory effects of Xiangmei Pills on the intestinal flora structure of rats with UC. Pharmacodynamic results showed that compared with that of the blank group, the colon tissue of the model group was congested, and ulcers were visible in the mucosa; compared with that in the model group, the histopathology of the colon of the rats with UC in the groups of Xiangmei Pills were improved, with scattered ulcers and reduced inflammatory cell infiltration. Chemical analysis showed that a total of 45 components were identified by mass spectrometry information, including 15 phenolic acids, 8 coumarins, 15 organic acids, 3 amino acids, 2 flavonoids, and 2 other components. Compared with those in the blank group, the levels of Ah R, CYP1A1, MUC-2, and IL-22 proteins in the colon tissue of rats in the model group were significantly decreased, and the level of IFN-γ protein was significantly increased; the intestinal flora of rats in the model group was disorganized, with a decrease in the abundance of the flora; the relative abundance of Bacteroidetes,unclassified genera of Ascomycetes, Prevotella of the Prevotella family, and Prevotella decreased significantly, and that of Firmicutes decreased, but the difference was not statistically significant. The relative abundance of Bacteroidetes, Bifidobacterium, and Lactobacillus increased significantly. Compared with those of the model group, the levels of Ah R, CYP1A1, MUC-2, and IL-22proteins in the colonic tissue of the groups of Xiangmei Pills were significantly higher, and the levels of IFN-γ proteins were significantly lower. The recovery of the intestinal flora was accelerated, and the diversity of the intestinal flora was significantly increased. The relative abundance of Bacteroidetes was significantly increased, and that of unclassified genera of Ascomycetes,Lactobacillus, Prevotella of the Prevotella family, and Prevotella was significantly increased. The relative abundance of Bacteroidetes and Bifidobacterium was significantly decreased. This study demonstrated that Xiangmei Pills can effectively treat UC, mainly through the phenolic acid and organic acid components to stimulate the intestinal barrier, regulate protein expression and the relative abundance and diversity of intestinal flora, and play a role in the treatment of UC.
Animals ; Colitis, Ulcerative/metabolism* ; Drugs, Chinese Herbal/chemistry* ; Rats, Sprague-Dawley ; Male ; Rats ; Gastrointestinal Microbiome/genetics* ; Chromatography, High Pressure Liquid ; Humans ; Mass Spectrometry ; RNA, Ribosomal, 16S/genetics* ; Bacteria/drug effects*

Based on the ultra performance liquid chromatography-quadrupole Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) technology, combined with related literature, databases, and reference material information, this study qualitatively analyzed the components of Dayuanyin in the tissue of rats after gavage and employed molecular docking technology to predict the rationality of the mechanism behind the antipyretic effect of the in vivo components in Dayuanyin. A total of 21, 26, 20, 21, 14, and 31 prototype components and 3, 16, 3, 7, 5, and 24 metabolites were identified from the heart, liver, spleen, lung, kidney, and hypothalamus of the rats, respectively, and the binding ability of key components and targets was further verified by molecular docking. The results showed that all components had good binding ability with targets. The established UPLC-Q-Exactive Orbitrap-MS could effectively and quickly identify the Dayuanyin components distributed in tissue and preliminarily identify their metabolites. Many components were identified in the hypothalamus, which suggested that the components delivered to the brain should be focused on in the study on Dayuanyin in the treatment of febrile diseases. The molecular docking technology was used to predict the rationality of the mechanism behind its antipyretic effect, which lays the foundation for the clarification of the material basis and action mechanism of Dayuanyin, the development of new preparations, and the prediction of quality markers.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Molecular Docking Simulation ; Male ; Antipyretics/metabolism* ; Rats, Sprague-Dawley ; Tissue Distribution ; Mass Spectrometry ; Chromatography, High Pressure Liquid ; Hypothalamus/metabolism*

Gastrointestinal (GI) cancers are a leading cause of cancer morbidity and mortality worldwide. Despite advances in treatment, cancer relapse remains a significant challenge, necessitating novel therapeutic strategies. In this study, we engineered nanobody-based chimeric antigen receptor (CAR) natural killer (NK) cells targeting cadherin 17 (CDH17) for the treatment of GI tumors. In addition, to enhance the efficacy of CAR-NK cells, we also incorporated CV1, a CD47-SIRPα axis inhibitor, to evaluate the anti-tumor effect of this combination. We found that CDH17-CAR-NK cells effectively eliminated GI cancers cells in a CDH17-dependent manner. CDH17-CAR-NK cells also exhibit potent in vivo anti-tumor effects in cancer cell-derived xenograft and patient-derived xenograft mouse models. Additionally, the anti-tumor activity of CDH17-CAR-NK cells is synergistically enhanced by CD47-signal regulatory protein α (SIRPα) axis inhibitor CV1, likely through augmented macrophages activation and an increase in M1-phenotype macrophages in the tumor microenvironment. Collectively, our findings suggest that CDH17-targeting CAR-NK cells are a promising strategy for GI cancers. The combination of CDH17-CAR-NK cells with CV1 emerges as a potential combinatorial approach to overcome the limitations of CAR-NK therapy. Further investigations are warranted to speed up the clinical translation of these findings.

ObjectiveTo study the effect of Tanreqing injection （TRQ） on the pulmonary flora in the rat model of chronic obstructive pulmonary disease （COPD）. MethodWistar rats were randomized into control， model， and TRQ groups. The rats in other groups except the control group were treated by smoking combined with intratracheal instillation of lipopolysaccharide for the modeling of COPD. The TRQ group was intraperitoneally injected with TRQ （2 g·kg^-1）. At the end of the experiment， after blood collection from the abdominal aorta of the rats， the lung tissue was collected for hematoxylin-eosin and picric sirius red staining to reveal the pathological changes. The lung lavage fluid was collected， and the diversity and relative abundance of lung flora in different groups were analyzed by 16S rDNA amplicon sequencing. ResultThe lungs of the control group were normal， and those of the model group showed neutrophil infiltration， telangiectasia， lung hemorrhage and emphysema in individual cases， and thickening of collagen fibers in the trachea. Compared with the model group， the TRQ group showed significantly improved lungs and recovered collagen fibers. The MLI analysis showed that compared with the control group， the model group showcased increased alveolar space （P<0.01）， which was reduced in the TRQ group （P<0.05）. Compared with the control group， the model group showed increased wall thickness （P<0.01）， and the increase was attenuated in the TRQ group （P<0.01）. TRQ increased the Simpson index and altered the α diversity of pulmonary flora. The results of principal co-ordinate analysis showed that TRQ changed the β diversity and reduced the β diversity index of pulmonary flora. At the genus level， the model group showed increased relative abundance of g_Bacillus and g_Brevundimonas and decreased relative abundance of g_Pseudomonas， compared with the control group. After treatment with TRQ， the relative abundance of g_Stenotrophomonas increased， and that of g_Bacillus decreased. The LEfSe of differential taxa between groups showed that the modeling increased the relative abundance of g_Lachnospiraceae_NK4A136_group， and TRQ treatment increased the relative abundance of g_Rhodococcus and g_Stenotrophomonas. ConclusionTRQ can regulate the diversity of pulmonary flora and restore the balance of bacterial genera in the rat model of COPD， which may be one of the mechanisms of the prevention and treatment of COPD with TRQ.

Endo-beta-N-acetylglucosaminidase (ENGase) is widely distributed in various organisms. The first reported ENGase activity was detected in Diplococcus pneumoniae in 1971. The protein (Endo D) was purified and its peptide sequence was determined in 1974. Three ENGases (Endo F1-F3) were discovered in Flavobacterium meningosepticum from 1982 to 1993. After that, the activity was detected from different species of bacteria, yeast, fungal, plant, mice, human, etc. Multiple ENGases were detected in some species, such as Arabidopsis thaliana and Trichoderma atroviride. The first preliminary crystallographic analysis of ENGase was conducted in 1994. But to date, only a few ENGases structures have been obtained, and the structure of human ENGase is still missing. The currently identified ENGases were distributed in the GH18 or GH85 families in Carbohydrate-Active enZyme (CAZy) database. GH18 ENGase only has hydrolytic activity, but GH85 ENGase has both hydrolytic and transglycosylation activity. Although ENGases of the two families have similar (β/α)8-TIM barrel structures, the active sites are slightly different. ENGase is an effective tool for glycan detection andglycan editing. Biochemically, ENGase can specifically hydrolyze β‑1,4 glycosidic bond between the twoN-acetylglucosamines (GlcNAc) on core pentasaccharide presented on glycopeptides and/or glycoproteins. Different ENGases may have different substrate specificity. The hydrolysis products are oligosaccharide chains and a GlcNAc or glycopeptides or glycoproteins with a GlcNAc. Conditionally, it can use the two products to produce a new glycopeptides or glycoprotein. Although ENGase is a common presentation in cell, its biological function remains unclear. Accumulated evidences demonstrated that ENGase is a none essential gene for living and a key regulator for differentiation. No ENGase gene was detected in the genomes of Saccharomyces cerevisiae and three other yeast species. Its expression was extremely low in lung. As glycoproteins are not produced by prokaryotic cells, a role for nutrition and/or microbial-host interaction was predicted for bacterium produced enzymes. In the embryonic lethality phenotype of the Ngly1-deficient mice can be partially rescued by Engase knockout, suggesting down regulation of Engase might be a solution for stress induced adaptation. Potential impacts of ENGase regulation on health and disease were presented. Rabeprazole, a drug used for stomach pain as a proton inhibitor, was identified as an inhibitor for ENGase. ENGases have been applied in vitro to produce antibodies with a designated glycan. The two step reactions were achieved by a pair of ENGase dominated for hydrolysis of substrate glycoprotein and synthesis of new glycoprotein with a free glycan of designed structure, respectively. In addition, ENGase was also been used in cell surface glycan editing. New application scenarios and new detection methods for glycobiological engineering are quickly opened up by the two functions of ENGase, especially in antibody remodeling and antibody drug conjugates. The discovery, distribution, structure property, enzymatic characteristics and recent researches in topical model organisms of ENGase were reviewed in this paper. Possible biological functions and mechanisms of ENGase, including differentiation, digestion of glycoproteins for nutrition and stress responding were hypothesised. In addition, the role of ENGase in glycan editing and synthetic biology was discussed. We hope this paper may provide insights for ENGase research and lay a solid foundation for applied and translational glycomics.

Objective To investigate the effect of ethanolic extract of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profiles in high-fat-diet-induced obese mice.Methods Twenty-four 6-week-old C57 BL/6 male mice were divided randomly into normal,model,drug-administration,and metformin groups.Mice in the normal group were fed a regular diet and mice in the other three groups were given high-fat diets.The drug-administration group was gavaged with 10 mL/kg ethanol extract of Cichorium glandulosum Boiss.et Huet.daily,and the metformin group was gavaged with 10 mL/kg metformin daily.After 10 weeks,livers were collected to measure hepatic total triglycerides(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C),and high-density lipoprotein(HDL)-C.Feces were collected and analyzed.Results Body weight(P＜0.0001),liver TG(P＜0.05),and TC(P＞0.05)were all significantly higher in model mice compared with normal mice,while LDL-C(P＞0.05)and HDL-C(P＜0.001)were significantly lower,indicating abnormal weight gain and lipid metabolism.Alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly reduced body weight(P＜0.0001),liver TG(P＜0.0001),serum TG(P＜0.05),TC(P＜0.01),and LDL-C(P＜0.05)in mice.Methodsological validation showed that the current method could accurately quantify 52 bile acids in feces.Analysis of the concentration of each type of bile acid revealed that alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly increased the secondary/primary bile acid ratio(P＜0.05).Multivariate analysis showed that the bile acid metabolic pattern was significantly altered in all groups.Eight differential bile acids were screened in the drug-administration group relative to the model group using variable importance of projection＞1 and P＜0.05.A search of the Kyoto Encyclopedia of Genes and Genomes database revealed that the differential bile acids were mainly involved in the secondary bile acid biosynthesis pathway.Correlation analysis showed that four differential bile acids,deoxycholic acid(r,=0.6445,P＜0.001),isolithocholic acid(r,=0.5879,P＜0.01),3β-deoxycholic acid(r,=0.6649,P＜0.001),and ω-rhamnoglutaric acid(rs=0.5387,P＜0.01),in feces were strongly positively correlated with body weight.Conclusions Cichorium glandulosum Boiss.et Huet.alcoholic extract may play a role in weight reduction and amelioration of dyslipidemia by modulating secondary bile acid biosynthesis and altering fecal bile acid metabolic profiles.

Objective:To compare the clinical efficacy of the H-Loop knotless double-row technique and the suture bridge technique in repairing L-shaped rotator cuff tears under arthroscopy.Methods:A retrospective analysis was performed on 58 patients with L-shaped rotator cuff injuries who underwent arthroscopic repair at Sun Yat-sen Memorial Hospital, Sun Yat-sen University, between January 2019 and December 2021. The H-Loop knotless double-row technique was used in 16 cases (8 males and 8 females, mean age 63.69±8.78 years), while the suture bridge technique was used in 42 cases (24 males and 18 females, mean age 61.02±7.02 years). The American Shoulder and Elbow Surgeons (ASES) score, University of California Los Angeles Shoulder Score (UCLA), Simple Shoulder Test (SST), shoulder range of motion, and muscle strength were evaluated and compared between the two groups one year after surgery.Results:The follow-up period was 12.81±0.98 months for the H-Loop group and 13.29±0.94 months for the suture bridge group. No significant differences were found between the groups in terms of age, sex, dominant hand, preoperative symptom duration, tear shape, tear size, or long head tendon amputation (P>0.05). The operative time was significantly shorter in the H-Loop group 67.50±16.02 minutes compared to the suture bridge group 76.67±13.19 minutes ( t=2.234, P=0.031). Additionally, the number of anchors used was significantly lower in the H-Loop group 2.00±0 compared to the suture bridge group 4.14±0.35 ( t=16.573, P<0.001). The ASES scores increased significantly in both groups: from 57.44±15.91 to 92.00±4.41 in the H-Loop group and from 58.21±16.58 to 87.71±6.19 in the suture bridge group ( F=53.439, P<0.001; F=72.511, P<0.001). Similarly, the UCLA scores improved from 20.63±3.79 to 31.56±3.65 in the H-Loop group and from 20.83±5.78 to 30.36±4.71 in the suture bridge group ( F=57.788, P<0.001; F=50.043, P<0.001). The Constant-Murley scores also showed significant improvement: from 68.50±15.31 to 87.5±8.70 in the H-Loop group and from 66.21±16.51 to 86.33±9.14 in the suture bridge group ( F=6.733, P<0.001; F=30.173, P<0.001). SST scores increased from 6.38±3.76 to 9.06±2.59 in the H-Loop group and from 6.55±3.31 to 9.17±2.45 in the suture bridge group ( F=2.847, P<0.001; F=11.096, P<0.001). The shoulder flexion range of motion increased from 158.75°±21.25° to 178.75°±47.07° in the H-Loop group and from 139.29°±45.12° to 179.76°±3.42° in the suture bridge group ( t=3.814, P=0.002; t=5.877, P<0.001). Shoulder abduction motion increased from 145°±45.46° to 178.75°±3.42° in the H-Loop group and from 135.24°±47.07° to 179.76°±1.54° in the suture bridge group ( t=2.952, P=0.001; t=6.185, P<0.001). Muscle strength improved from 53.36±25.21 N to 73.69±24.09 N in the H-Loop group and from 43.31±24.49 N to 61.8±30.07 N in the suture bridge group ( t=4.916, P<0.001; t=5.623, P<0.001). The ASES score at one year post-surgery was significantly higher in the H-Loop group 92.00±4.41 compared to the suture bridge group 87.71±6.19 ( t=2.529, P=0.014). There were no significant differences in UCLA scores, Constant-Murley scores, SST scores, shoulder motion, or muscle strength between the groups ( P>0.05). Conclusion:The H-Loop technique provides a good early curative effect. Compared to the traditional suture bridge technique, the H-Loop technique offers a higher early postoperative ASES score, shorter operative time, and fewer anchors required.

Objective To investigate the application effect of a bedside ultrasound-guided intestinal cleaning program in patients with severe acute pancreatitis.Methods A total of 51 patients with severe acute pancreatitis admitted to the ICU of a tertiary A hospital in Shandong from March to September 2023 were selected by convenience sampling method,and they were divided into an experimental group and a control group according to random number table method.The experimental group was given the bedside ultrasound-guided intestinal cleaning program,and the control group was given the routine intestinal cleaning program.Acute gastrointestinal injury ultrasonography score,the incidence of grade Ⅲ acute gastrointestinal injury and intra-abdominal pressure were compared between the 2 groups before intervention,on the 3rd and 5th day.Results There was an interaction effect between time and group in the comparison of acute gastrointestinal injury ultrasonography scores in the 2 groups(F=7.478,P<0.001);simple effect analysis showed that acute gastrointestinal injury ultrasonography scores in the experimental group were lower than those in control group on the 3rd and 5th day,with statistically significant differences(P<0.05).The incidence of grade Ⅲ acute gastrointestinal injury in the experimental group(23%)was lower than that in the control group(60%),with statistically significant differences(P<0.05).The intra-abdominal pressure had an interaction effect between the 2 groups(F=47.128,P<0.001);simple effect analysis showed that the intra-abdominal pressure in the experimental group was lower than that in the control group on the 3rd and 5th day,with statistically significant differences(P<0.05).Conclusion The bedside ultrasound-guided intestinal cleaning program can improve acute gastrointestinal injury and reduce intra-abdominal hypertension in patients with severe acute pancreatitis.