Screening carbonyl reductases with the ability to catalyze the reduction of complex carbonyl compounds is of great significance for the biosynthesis of R-tolvaptan(R-TVP). In this study, the target carbonyl reductase in the crude enzyme extract of rabbit liver was separated, purified, and identified by ammonium sulfate precipitation, gel-filtration chromatography, ion exchange chromatography, affinity chromatography, and protein mass spectrometry. With the rabbit liver genome as the template, the gene encoding the carbonyl reductase rlsr5 was amplified by PCR and the recombinant strain was successfully constructed. After RLSR5 was purified by affinity chromatography, its enzymatic properties were characterized. The results indicated that the gene sequence of rlsr5 was 972 bp, encoding a protein with a molecular weight of 40 kDa. RLSR5 was a dimeric protein, and each monomer was composed of a (α/β)₈-barrel structure. RLSR5 could asymmetrically reduce 7-chloro-1-[2-methyl-4-[(2- methylbenzoyl)amino]benzoyl]-5-oxo-2,3,4,5-tetrahydro-1H-1-benzazepine (prochiral ketone, PK) to synthesize R-TVP. The specific activity of the enzyme was 36.64 U/mg, and the optical purity of the product was 99%. This enzyme showcased the optimal performance at pH 6.0 and 30 °C. It was independent of metal ions, with the activity enhanced by Mn²⁺. This study lays a foundation for the biosynthesis of tolvaptan of optical grade.
Animals ; Rabbits ; Alcohol Oxidoreductases/biosynthesis* ; Recombinant Proteins/metabolism* ; Escherichia coli/metabolism* ; Liver/enzymology*

Objective To analyze dynamic contrast-enhanced MRI (DCE-MRI) radiomic features using machine learning algorithms, and to develop and validate a predictive model for HER-2 status in breast cancer. Methods The DCE-MRI images of 272 treatment-naive female patients with breast cancer between 2020 and 2022 were included in this study. Regions of interest (ROIs) were manually segmented using 3d-Slicer software, and radiomic features were extracted. All patients were randomly divided into training sets or validation sets at a ratio of 4∶1. The least absolute shrinkage and selection operator (LASSO) algorithm was used for feature screening on the training set, followed by the development of predictive models using six machine learning algorithms. Internal cross-validation was performed to compare the performance differences between the models. The best-performing model was selected, trained on the training set, and evaluated on the validation set. Evaluation metrics included area under the curve (AUC), sensitivity, specificity, precision, and recall rate. Results The clinical data of patients in the training set and validation set showed no significant differences. Five features were identified by the LASSO algorithm. With these features, six machine learning models were developed on the training set, and their predictive performance was internally cross-validated using the bagging method. XGBoost model had the highest mean AUC (0.696), followed by RF model (0.690); XGBoost model had the highest mean precision (0.756), followed by LR and RF models. Therefore, XGBoost was the optimal model. An HER-2 predictive model was built using the XGBoost algorithm on the training set and applied to the validation set. The AUC, precision, sensitivity, and specificity of the predictive model on the validation set were calculated, and ROC curves, precision-recall curves, calibration curves, and decision-making curves were plotted. Conclusion This study constructed and evaluated different DCE-MRI radiomics-based machine learning models for predicting HER-2 status in breast cancer. Among them, XGBoost algorithm performed the best and has the potential to become a new non-invasive method for preoperative prediction of HER-2 status, providing reliable evidence for personalized clinical diagnosis and treatment.

With reference to the Information and Documentation-Resource Description (GB/T 3792-2021) and Bibliographical Description for Ancient Chinese Books (GB/T 3792.7-2008) and other cataloging standards and rules, drawing on the practical experience of cataloging ancient TCM books, Bibliographical Cataloging for Ancient TCM Books was formulated. This standard specifies the entry items and their order of ancient TCM books, cataloging identifier, cataloging text, cataloging information source, and cataloging item details. The standard can provide standardized and unified guiding principles and methods for the work of ancient TCM books, and promote the sharing and utilization of ancient TCM books.

Objective To explore the effects of plantamajoside on the proliferation and invasion of human gastric cancer cell line BGC823.Methods BGC823 cells were randomly separated into a control group,a plantamajoside group,an AAV-NC(transfection of empty plasmids packaged with lentivirus)group and a plantamajoside+AAV-HIF-1α(transfection of HIF-1α overexpression plasmid packaged with lentivirus)group.Cell proliferation,inva-sion,apoptosis,the numbers of vascular mimicry(VM)lumens and vascular branches,the expression of prolifera-tion,apoptosis,epithelial mesenchymal transition(EMT)related proteins,HIF-1α/VEGF pathway proteins of cells were all examined.Results Compared with control group,the BGC823 cell viability,colony formation num-ber,invasion number,VM lumen number,vascular branch number,and expression of Ki-67,PCNA,vimentin,MMP9,Snail,VEGFA,VE-cadhering,HIF-1α and VEGF protein were all lower in plantamajoside group(P＜0.05).The apoptosis rate,the cleaved Caspase-3,Bax,and E-cadherin protein expression were significantly increased(P＜0.05).Compared with plantamajoside group,the BGC823 cell viability,colony formation number,invasion number,VM lumen number,vascular branch number,and expression of Ki-67,PCNA,vimentin,MMP9,Snail,VEGFA,VE-cadherin,HIF-1α and VEGF protein were higher in the plantamajoside+AAV-HIF-1α group(P＜0.05).The apoptosis rate,the cleaved caspase-3,Bax,and E-cadherin protein expression were lower(P＜0.05).Conclusions Plantamajoside inhibits proliferation,EMT,invasion,and VM of human gastric cancer cell line and induce its apoptosis.

Alzheimer's disease(AD)is the leading cause of dementia,and no effective treatment has been devel-oped for it thus far.Recently,the use of natural compounds in the treatment of neurodegenerative diseases has garnered significant attention owing to their minimal adverse reactions.Accordingly,the potential therapeutic effect of pterostilbene(PTS)on AD has been demonstrated in multiple in vivo and in vitro experiments.In this study,we systematically reviewed and summarized the results of these studies investigating the use of PTS for treating AD.Analysis of the literature revealed that PTS may play a role in AD treatment through various mechanisms,including anti-oxidative damage,anti-neuroinflammation,anti-apoptosis,cholinesterase activity inhibition,attenuation of β-amyloid deposi-tion,and tau protein hyperphosphorylation.Moreover,PTS interferes with the progression of AD by regulating the activities of peroxisome proliferator-activated receptor alpha(PPAR-α),monoamine oxi-dase B(MAO-B),silent information regulator sirtuin 1(SIRT1),and phosphodiesterase 4A(PDE4A).Furthermore,to further elucidate the potential therapeutic mechanisms of PTS in AD,we employed network pharmacology and molecular docking technology to perform molecular docking of related proteins,and the obtained binding energies ranged from-2.83 to-5.14 kj/mol,indicating that these proteins exhibit good binding ability with PTS.Network pharmacology analysis revealed multiple po-tential mechanisms of action for PTS in AD.In summary,by systematically collating and summarizing the relevant studies on the role of PTS in treatment of AD,it is anticipated that this will serve as a reference for the precise targeted prevention and treatment of AD,either using PTS or other developed drug interventions.

The thought of the way of heaven is the core philosophy of the Book of Changes， aiming to achieve the “pursuing good fortune and avoiding disaster” of human affairs or “seeking benefits and avoiding harm” in technology. The embedded value orientations， such as “continuous life”， “utmost sincerity”， and “correct desire”， provide ethical considerations for “seeking benefits and avoiding harm” in human gene editing technology. Among them， the thought of “continuous life” affirms the potential value of gene editing technology for human reproduction， diagnosis and treatment， or prevention of diseases and disabilities from two aspects of human reproduction and individual body and mind， while adopting a cautious attitude towards the risks of technology. The thought of “utmost sincerity” advocates that gene editing should be studied and applied following objective laws. The thought of “correct desire” puts forward the norm of human desire in human gene editing. In terms of the issue of offspring autonomy， the thought of the way of heaven provides some ethical support for parents to implement gene editing for their offspring for the sake of the offspring’s physical and mental development. In conclusion， the application of the thought of the way of heaven in the Book of Changes hopes that human gene editing technology will benefit mankind and provide it with broader prospects and a better future.

Objective To explore differentially expressed proteins in the serum exosomes of acute myeloid leukemia(AML)patients and healthy controls by using the proteomics method,and provide new biomarker for the diagnosis and treatment of AML.Methods A total of 76 AML patients diagnosed and treated in the First People's Hospital of Honghe State from November 2022 to June 2024 were selected as the experimental group,and 60 healthy physical examination participants were selected as the control group.Tandem Mass Tag(TMT)labeled proteomics was used to identify the differences in protein expression in serum exosomes between the experimental and control groups,and the identified differentially expressed proteins were subjected to bioinformatics analysis.Western-Blot was used to verify the differentially expressed proteins.Results Comparedwith healthy controls,146 aberrantly expressed proteinswere detected in serum exosomes of AML patients,of which 89 were up-regulated and 57 were down-regulated.Among them,Alpha-1-antichymotrypsin(α1-ACT),Angiogenin(ANG),Vitronectin(VIT),Clusterin(Clu),Fibronectin(FN),Keratin type I cytoskeletal-18(KRT18),and actinin alpha4(ACTN4)showed changed significantly.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis indicated that differentially expresseproteins were involved in biological processes such as neutrophil immunity,neutrophil degranulation,neutrophil activation immune response,and protein activation cascade,significantly enriched in 25 biological pathways including complement and coagulation cascades,extracellular matrix(ECM)-receptor interaction,and hypoxia-inducible factor-1(HIF-1)signaling pathways.Western-blot validation results showed that compared with the control group,the expression levels of ANG,Clu and FN in the serum exosomes of AML patients were significantly increased,while the expression level of ACTN4 was significantly decreased,with statistically significant differences(t=-11.854～18.569,all P<0.05).The expression levels of ANG and FN were correlated with white blood cell count(F=8.888,7.818,all P<0.05)and cytogenetics(F=8.619,7.983,P<0.05).The expression level of Clu protein was correlated with white blood cell count(F=2.571,P<0.05),but not significantly associated with cytogenetics(F=1.886,P>0.05).ANG,FN and Clu showed no significant correlation with FAB classification(F=0.175,0.434,0.042,all P>0.05).Conclusion The expression of exosomal proteins in the serum of AML patients compared to healthy individuals are significant differences,which may serve as serological markers for the diagnosis and treatment of AML.

Objective To investigate the expression changes of serum circRNA membrane bound O-acyltransferase 2(circRNA MBOAT2)and circRNA recombinant actinin 4(circRNA ACTN4)in patients with cholangiocarcinoma and their relationship with prognosis.Methods 96 patients with cholangiocarcinoma treated at the First Hospital of Handan City from January 2020 to January 2022 in Department of Hepatobiliary Surgery were regarded as as the cholangiocarcinoma group.Additionally,85 patients with intraductal stones and 90 healthy volunteers were collected as the stone group and control group,respectively.Quantitative real-time reverse transcription PCR(qRT-PCR)was applied to detect the expression levels of circRNA MBOAT2 and circRNA ACTN4.χ2 test was applied to analyze the relationship between circRNA MBOAT2 and circRNA ACTN4 and clinical pathological features.Pearson was applied to analyze the correlation between circRNA MBOAT2 and circRNA ACTN4 in patients with cholangiocarcinoma.ROC was applied to analyze the diagnostic value of circRNA MBOAT2 and circRNA ACTN4 in the occurrence of cholangiocarcinoma.COX was applied to analyze the influencing factors of poor prognosis in patients with cholangiocarcinoma.Kaplan-Meier method was applied for survival analysis.Results circRNA MBOAT2(1.24±0.38)and circRNA ACTN4(1.27±0.42)in cholangiocarcinoma group were higher than those in stone group(1.02±0.31,1.05±0.34)and control group(0.83±0.24,0.78±0.21),and the stone group was higher than that in control group,with statistically significance(t=5.016～14.025,all P<0.05).There was a positive correlation between circRNA MBOAT2 and circRNA ACTN4 in patients with cholangiocarcinoma group(r=0.428,P<0.05).Combined diagnosis of MBOAT2 and ACTN4 was better than single diagnosis of cholangiocarcinoma(Z=4.063,4.004,all P<0.05);circRNA MBOAT2 and circRNA ACTN4 were correlated with clinical staging and lymph node status(t=5.091～5.984,all P<0.05).Positive lymph node status and elevated levels of circRNA MBOAT2 and circRNA ACTN4 were independent risk factors for mortality(HR=1.527,1.582,1.727,all P<0.05).The cumulative survival rate of patients with high expression of circRNA MBOAT2(23.40%vs 46.94%)and circRNA ACTN4(24.00%vs 47.83%)was lower than that of patients with low expression(χ2=5.809,5.946,all P<0.05).Conclusion The serum levels of circRNA MBOAT2 and circRNA ACTN4 increase with the progression of the disease,and the combination of the two has certain value in diagnosing the occurrence of cholangiocarcinoma.

Alzheimer's disease (AD) is the leading cause of dementia, and no effective treatment has been developed for it thus far. Recently, the use of natural compounds in the treatment of neurodegenerative diseases has garnered significant attention owing to their minimal adverse reactions. Accordingly, the potential therapeutic effect of pterostilbene (PTS) on AD has been demonstrated in multiple in vivo and in vitro experiments. In this study, we systematically reviewed and summarized the results of these studies investigating the use of PTS for treating AD. Analysis of the literature revealed that PTS may play a role in AD treatment through various mechanisms, including anti-oxidative damage, anti-neuroinflammation, anti-apoptosis, cholinesterase activity inhibition, attenuation of β-amyloid deposition, and tau protein hyperphosphorylation. Moreover, PTS interferes with the progression of AD by regulating the activities of peroxisome proliferator-activated receptor alpha (PPAR-α), monoamine oxidase B (MAO-B), silent information regulator sirtuin 1 (SIRT1), and phosphodiesterase 4A (PDE4A). Furthermore, to further elucidate the potential therapeutic mechanisms of PTS in AD, we employed network pharmacology and molecular docking technology to perform molecular docking of related proteins, and the obtained binding energies ranged from -2.83 to -5.14 kJ/mol, indicating that these proteins exhibit good binding ability with PTS. Network pharmacology analysis revealed multiple potential mechanisms of action for PTS in AD. In summary, by systematically collating and summarizing the relevant studies on the role of PTS in treatment of AD, it is anticipated that this will serve as a reference for the precise targeted prevention and treatment of AD, either using PTS or other developed drug interventions.

Objective:To investigate the role of the IgG subtypes (IgG1 and IgG2a) in anti-glycoprotein (GP) Ⅰbα antibody-induced platelet clearance.Methods:Venous blood was collected from healthy volunteers, and platelets were separated. The phagocytosis of human platelets by human acute monocytic leukemia cells (THP-1 cells) induced by different anti-GPⅠbα antibodies (AN51, AK2, HIP1, TM60, VM16d, WM23, and SZ2) was detected by flow cytometry. The effects of the AN51 full-length antibody, F (ab') 2, and Fab fragments on platelet phagocytosis by THP-1 cells were detected by flow cytometry. Then, the Fc blocking antibody 2.4G2 and normal rat IgG2a or IgG1 were injected into C57BL/6J mice via the posterior ocular vein, and their effects on platelet reduction induced by R300 were detected by a hematology analyzer. Results:Compared with IgG1, the IgG2a subtype of anti-GPⅠbα antibodies induced the phagocytosis of platelets by THP-1 cells in vitro ( P<0.05). In contrast to the AN51 full-length antibody, neither AN51 F (ab') 2 nor the Fab fragment could induce THP-1 cells to phagocytose platelets ( P<0.05). Compared with the control group, anti-mouse GPⅠbα R300-induced thrombocytopenia in mice was reduced at 2, 4, and 6 h after the injection of Fc blocking antibody 2.4G2 ( P<0.05). Similarly, R300-induced thrombocytopenia in mice was reduced at 2, 4, and 6 h after the injection of rat IgG2a ( P<0.05) . Conclusion:IgG2a plays an important role in anti-GPⅠbα-induced clearance.