Rheumatoid arthritis belongs to arthralgia in the theory of traditional Chinese medicine, with cold dampness obstruction pattern (CDO) and damp heat obstruction pattern (DHO) as the main pattern types. Fine therapeutic effects have been obtained in clinical practice following the differentiation of CDO and DHO. However, suitable animal models are not available currently. For adapting to the clinical diagnosis, as well as carrying out basic research of integrated Chinese and Western medicine and preclinical study on new Chinese medicine in a better way, the “Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold Dampness Obstruction Pattern and Damp Heat Obstruction Pattern” (referred to as “Guidelines”) were compiled by our research group in cooperation with renowned experts in clinical, pharmaceutical, zoological, and methodological research fields. According to the theory of disease and syndrome integration, experts standardized the establishment methods for animal models of rheumatoid arthritis with CDO and DHO. The Guidelines were compiled via the nominal group method according to the principle of evidence coming in main place, consensus as the auxiliary, and experience as the references. Contents such as animal species, arthritis induction methods, external stimulation conditions, and assessment indicators were specified in the Guidelines based on the comprehensive evaluation of pathogenesis homology, behavioral phenotypic consistency, and drug therapy predictability between animal models and human diseases. The Guidelines will be applicable to the research on syndromes in rheumatoid arthritis, elucidation of acting mechanism, and development of new medicine, and provide reference for standardizing other types of animal models with the integration of disease and syndrome, to promote the modernization of Chinese medicine research.

Beta-amylases (BAMs), key enzymes in starch hydrolysis, play an important role in plant growth, development, and resistance to abiotic stress. To mine the saline-alkali tolerance-related BAM genes in alfalfa (Medicago sativa L.), we identified MsBAM genes in the whole genome. The physicochemical properties, phylogeny, gene structures, conserved motifs, secondary structures, promoter cis-acting elements, chromosome localization, and gene replication relationships of BAM gene family members were analyzed. RNA-seq and quantitative real-time PCR (qRT-PCR) were employed to analyze the expression patterns of BAM family members under saline-alkali stress. The results showed that 54 BAM genes were identified in the genome, which were classified into 8 subgroups according to the phylogenetic tree. The members of the same subgroup had similar gene structures except that those of subgroups 1 and 7 had large differences. Conserved motif analysis showed that all MsBAM proteins had a typical glycohydrolysis domain. The chromosome localization analysis showed that MsBAM gene family members were unevenly distributed on 27 chromosomes. The duplication of gene segments led to the increase in BAM gene number in alfalfa. The promoters of BAM genes contained a large number of elements in response to plant hormones and stress. Transcriptome data and qRT-PCR results showed that the expression levels of most MsBAM genes were up-regulated in response to saline-alkali stress. Under the saline-alkali stress, the expression levels of 28 genes, including MsBAM6, were up-regulated on days 1 and 7, and those of 5 genes, including MsBAM9, were up-regulated by over 2 folds. In addition, under salt-alkali stress, BAM activity and soluble sugar content were significantly increased. These results indicate that BAM genes play a key role in alfalfa in response to saline-alkali stress, laying a foundation for further research in this field.
Medicago sativa/physiology* ; beta-Amylase/metabolism* ; Phylogeny ; Gene Expression Regulation, Plant ; Stress, Physiological/genetics* ; Multigene Family ; Alkalies ; Plant Proteins/genetics*

The organoid co-culture model, as a novel tool for recreating a three-dimensional microenvironment to study cell-cell interactions, has demonstrated significant application potential in biomedical research in recent years. By simulating the in vivo tissue microenvironment, this model provides a more precise experimental platform for investigating complex cellular interactions, particularly in areas such as tumor immune evasion mechanisms, drug sensitivity testing, and the pathological characterization of neurodegenerative diseases, where it has demonstrated significant value. However, the organoid co-culture model still faces several challenges in terms of standardized procedures, large-scale cultivation, ethical guidelines, and future development. In particular, in the field of laboratory animal science, how to effectively combine organoids with traditional animal models, and how to select the most appropriate model for different research needs while exploring its potential for replacement, remain pressing issues. In the context of ethical approval and the replacement of animal experiments, the organoid co-culture model offers an experimental approach that better aligns with the "3R" principle (Replacement, Reduction, Refinement), potentially becoming an important tool for replacing traditional animal models. To this end, this paper reviews the latest advances and key challenges in this field, providing a detailed description of the construction methods for organoid co-culture models and discussing their applications in disease mechanism research and drug screening. The paper also systematically compares the organoid co-culture models with traditional animal models, exploring the criteria for selecting the appropriate model for specific applications. Furthermore, this paper discusses the potential value of organoid co-culture models as alternatives to animal experiments and anticipates future development trends of this technology. Through these discussions, the paper aims to promote the innovation and development of organoid co-culture technology and provide new perspectives and scientific evidence for future research.

The organoid co-culture model, as a novel tool for recreating a three-dimensional microenvironment to study cell-cell interactions, has demonstrated significant application potential in biomedical research in recent years. By simulating the in vivo tissue microenvironment, this model provides a more precise experimental platform for investigating complex cellular interactions, particularly in areas such as tumor immune evasion mechanisms, drug sensitivity testing, and the pathological characterization of neurodegenerative diseases, where it has demonstrated significant value. However, the organoid co-culture model still faces several challenges in terms of standardized procedures, large-scale cultivation, ethical guidelines, and future development. In particular, in the field of laboratory animal science, how to effectively combine organoids with traditional animal models, and how to select the most appropriate model for different research needs while exploring its potential for replacement, remain pressing issues. In the context of ethical approval and the replacement of animal experiments, the organoid co-culture model offers an experimental approach that better aligns with the "3R" principle (Replacement, Reduction, Refinement), potentially becoming an important tool for replacing traditional animal models. To this end, this paper reviews the latest advances and key challenges in this field, providing a detailed description of the construction methods for organoid co-culture models and discussing their applications in disease mechanism research and drug screening. The paper also systematically compares the organoid co-culture models with traditional animal models, exploring the criteria for selecting the appropriate model for specific applications. Furthermore, this paper discusses the potential value of organoid co-culture models as alternatives to animal experiments and anticipates future development trends of this technology. Through these discussions, the paper aims to promote the innovation and development of organoid co-culture technology and provide new perspectives and scientific evidence for future research.

ObjectiveTo elucidate the chemical composition of Danshenyin and its blood components in rats after oral administration. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） coupled with PeakView 1.2 software was used to systematically characterize and identify the components of Danshenyin aqueous extract and its migratory components in rat blood after oral administration based on the retention time， quasi-molecular ion peaks， secondary fragmentation ions， and literature reports， and a preliminary compounds identification of Salviae Miltiorrhizae Radix et Rhizoma aqueous extract， the co-decoction of Santali Albi Lignum and Amomi Fructus was carried out to attribute the chemical constituents of the aqueous extract of Danshenyin. ResultsA total of 73 compounds， including 21 phenolic acids， 23 diterpenes， 6 flavonoids， 7 organic acids， 3 volatile oils and 13 others， were identified from the aqueous extract of Danshenyin. And 36 prototypes and 15 metabolites were identified in rat plasma， the major metabolic pathways included reduction， hydration， hydroxylation， demethylation， methylation， sulfation and others， these metabolites were mainly derived from tanshinones and salvianolic acids. ConclusionThe main blood components of the aqueous extract of Danshenyin are salvianolic acids and tanshinones， which may be the material basis of the efficacy. This study can provide reference for pharmacological research， quality control， and clinical application of Danshenyin.

Rheumatoid arthritis belongs to arthralgia syndrome in the theory of traditional Chinese medicine， and cold-dampness obstruction syndrome and dampness-heat obstruction syndrome are core syndromes and main syndrome differentiation types of this disease. Fine therapeutic effects have been obtained in the long-term clinical practice of many famous traditional Chinese medicine practitioners following the syndrome differentiation and treatment based on the guiding principles of cold and heat. To adapt to the clinical diagnosis practice of combining disease differentiation and syndrome differentiation， and to better carry out basic research on integrated Chinese and Western medicine and preclinical study on new traditional Chinese medicines， Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-Dampness Obstruction Syndrome and Dampness-Heat Obstruction Syndrome （hereinafter referred to as the Guidelines） were compiled by our research group， in cooperation with the renowned experts in research fields including traditional Chinese medicine， clinical medicine， zoology and evidence-based medicine， which provide a meaningful reference for scientific research， teaching and clinical applications. The compilation process of the Guidelines was guided by the theory of disease and syndrome integration and the principles of "evidence takes the main place， consensus plays an auxiliary role， and experience serves as the reference". Based on the comprehensive evaluation of pathogenesis homology， behavioral phenotypic consistency， and drug treatment predictability compared between animal models and human diseases， by the nominal group method， "recommendations" were formed for recommendations supported by evidence， and "consensus recommendations" were formed for recommendations not supported by evidence. Guidelines were formed involving content such as animal types， arthritis modeling methods， external stimulation conditions， and modeling assessment indicators during the establishment of the animal models of rheumatoid arthritis with cold-dampness obstruction syndrome and dampness-heat obstruction syndrome. The Guidelines are applicable for the disease and syndrome research on rheumatoid arthritis， investigation of therapeutic mechanisms， and development of new traditional Chinese medicine. The Guidelines also provide a reference for the establishment of guidelines on other types of diseases and syndromes combined with animal models to further promote the modernization of traditional Chinese medicine research and its integration with international academic development.

The Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome （hereinafter referred to as the Guidelines） （No. T/CACM1567-2024） was published by Chinese Association of Chinese Medicine on January 11， 2024. To assist researchers and medical workers in understanding and applying the Guidelines more accurately， and also to provide reference and assistance for the establishment of guidelines on other types of diseases and syndromes combined with animal models， this paper made a declaration of the workflow， technological links， development references， promotion of its application and after-effect evaluation of the Guidelines that has been made according to the requirements of "Draft Group Standard of the Standardization Office of the Chinese Association of Traditional Chinese Medicine".

Objective To analyze the causal relationship between brain resting state functional magnetic resonance imaging(rsfMRI)network and myocardial infarction using the Mendelian randomization(MR)research method.Methods Four analytical methods of MR[inverse-variance weighted(IVW),weighted median method(WME),weighted mode method,and MR-Egger method]were used to conduct a two-way two-sample analysis between 191 types of rsfMRI phenotypes of the brain(34 691 types)and myocardial infarction.And based on the aggregated analysis of genome-wide association studies(GWAS)data,MR was used to explain the causal relationship between the two.Then,the IVW method was used for analysis and research as the main result,and other analytical methods were used as supplementary analyses.Finally,the credibility of the results was verified by using sensitivity analysis.Results The neural connection activities located at the edge of the fusiform gyrus of the temporal lobe and the default mode network[odds ratio(OR)=1.260,95%confidence interval(95%CI)1.120-1.420,P<0.001]may increase the risk of myocardial infarction.In addition,the motor and subcortical cerebellar networks located in the posterior central gyrus,anterior central gyrus,and cerebellum(OR=0.770,95%CI was 0.680-0.880,P<0.001),and the subcortical cerebellar and motor networks located in the cerebellum and paracentric lobules(OR=0.810,95%CI was 0.720-0.900,P<0.001)and the neural connection activities of the motor and subcortical cerebellar networks located in the posterior central gyrus,anterior central gyrus and subcortex(OR=0.730,95%CI was 0.630-0.840,P<0.001)were associated with a reduced risk of myocardial infarction.Sensitivity analysis showed that the research results were reliable,and after the reverse MR study,it was confirmed that there was no reverse causal association between the two.Conclusion The network signals located at the edge of the fusiform gyrus of the temporal lobe and in the default mode may be potential risk factors for myocardial infarction;the motor and subcortical cerebellar networks located in the posterior central gyrus,anterior central gyrus and cerebellum,as well as the subcortical cerebellar and motor networks located in the cerebellum and paracentric lobules,and the neural connection activities of the motor and subcortical cerebellar networks located in the posterior central gyrus,anterior central gyrus and subcortex may be potential protective factors,providing new ideas for the prevention and treatment of myocardial infarction.

Objective To explore the impacts of remimazolam on intraoperative intracranial pressure(ICP)and cerebral oxygenation in patients undergoing laparoscopic lower abdominal and pelvic surgery under CO2 pneumoperitoneum combined with Trendelenburg position.Methods Eighty-eight patients scheduled to undergo laparoscopic lower abdominal and pelvic surgery were randomly assigned to the remimazolam group(n=44)and the sevoflurane group(n=44).In the remimazolam group,continuous infusion of remimazolam at a rate of 1 mg/(kg·h)was administered for anesthesia maintenance.In contrast,the sevoflurane group inhaled 2％sevoflurane.Heart rate(HR),mean arterial pressure(MAP),peak airway pressure(Peak),plateau airway pressure(Plat),end-tidal CO2(PETCO2),regional cerebral oxygen saturation(rSO2),and optic nerve sheath diameter(ONSD)of both eyes were measured and recorded at the following time points:prior to anesthesia induction(T0),5 minutes after induction(T1),10 minutes(T2),30 minutes(T3),and 60 minutes(T4)after the establishment of pneumo-peritoneum in Trendelenburg position,as well as 30 minutes after deflation in the supine position(T5).Results No significant intergroup differences were detected in HR,MAP,Peak,Plat,or PETCO2 at any time point(P>0.05).In both groups,Peak and Plat values were significantly higher at T2-T4 compared to T1(P<0.05).Regard-ing the ONSD)no intergroup differences were noted at T0 and T1(P>0.05).From T2 to T5,ONSD in both groups increased significantly relative to T0..It gradually rose with the prolongation of pneumoperitoneum and Trendelen-burg positioning and showed a slight decrease at T5(P<0.05).Specifically,at T3 and T4,the remimazolam group exhibited significantly smaller ONSD values than the sevoflurane group(P<0.05).Throughout the study,no inter-group differences in rSO2 were observed(P>0.05).Conclusion In laparoscopic lower abdominal and pelvic surgeries performed in the Trendelenburg position,intravenous anesthesia with remimazolam may be more effective in mitigating the elevation of intracranial pressure compared to sevoflurane inhalation.

Objective To investigate the clinical value of serum mast cell carboxypeptidase(MC-CP),C-C motif chemokine 26(CCL26),and decoy receptor 3(DcR3)in the diagnosis of obstructive sleep apnea syndrome(OSAS)in chronic obstructive pulmonary disease(COPD).Methods Ninety COPD patients who visited the First Affiliated Hospital of Hebei North University from January 2021 to January 2023 were collected.Among them,48 patients with simple COPD were included in the COPD group,and 42 patients with COPD combined with OSAS were included in the COPD-OSAS group.During the same period,48 healthy volunteers who underwent physical examination in that Hospital of Hebei North collected as the control group.Enzyme linked immunosorbent assay(ELISA)was applied to detect serum level of MC-CP,CCL26,and DcR3.Receiver operating characteristic(ROC)curve was applied to analyze the clinical value of serum level of MC-CP,CCL26,and DcR3 in the diag-nosis of COPD complicated with OSAS.Multivariate Logistic regression was applied to analyze the influencing fac-tors of COPD complicated with OSAS.Results Compared with the control group,the smoking index,C-reactive protein(CRP)and white blood cell count(WBC)in the COPD and COPD-OSAS groups increased obviously in sequence,the ratio of forced expiratory volume in first second to forced vital capacity(FEV1/FVC)decreased obviously in sequence(P<0.05);Compared with the control group,the level of MC-CP,CCL26,and DcR3 in patients with COPD and COPD-OSAS increased significantly in sequence(P<0.05);The combination of serum MC-CP,CCL26 and DcR3 had a higher area under the curve(AUC)for the diagnosis of COPD complicated with OSAS compared to the individual diagnosis(Z=4.066,P<0.001;Z=2.391,P<0.05;Z=2.353,P<0.05).Multivariate Logistic regression analysis showed that smoking index,serum level of MC-CP,CCL26 and DcR3 were influencing factors for COPD complicated with OSAS(P<0.05).Conclusions The simultane-ously increased expression of MC-CP,CCL26 and DcR3 in the serum of COPD may support clinical diagnostic of COPD patients with OSAS.