Traditional Chinese Medicine (TCM), as a treasure of the Chinese nation, plays a significant role in maintaining public health. In 2019, the Central Committee of the Communist Party of China and the State Council proposed for the first time the establishment of a TCM registration and evaluation evidence system that integrates TCM theory, "personal experience" and clinical trials (referred to as the "Three-in-One" System) to promote the inheritance and innovation of TCM. Subsequently, the National Medical Products Administration issued several guiding principles to advance the improvement and implementation of this system. Owing to the complexity of its implementation, there are still differing understandings within the TCM industry regarding the positioning of the "Three-in-One" Registration and Evaluation Evidence System, as well as the connotation and value orientation of the "personal experience." To address this, Academician WANG Qi, President of the TCM Association, China International Exchange and Promotion Association for Medical and Healthcare and TCM master, led a group of academicians, TCM masters, TCM pharmacology experts and clinical TCM experts to convene a "Seminar on Promoting the Implementation of the ′Three-in-One′ Registration and Evaluation Evidence System for Chinese Medicinals." Through extensive discussions, an expert consensus was formed, clarifying the different roles of the TCM theory, "personal experience" and clinical trials within the system. It was further emphasized that the "personal experience" is the core of this system, and its data should be derived from clinical practice scenarios. In the future, the improvement of this system will require collaborative efforts across multiple fields to promote the high-quality development of the Chinese medicinal industry.

Aim To explore the effect of serum contai-ning Duhuo Jisheng Decoction on the proliferation,ap-optosis and inflammation of fibroblast-like synoviocytes(FLS)induced by tumor necrosis factor-α(TNF-α)in rheumatoid arthritis(RA)and to reveal the under-lying mechanism.Methods The MH7A cells were divided into five groups:normal group(10％blank se-rum),model group(10 μg·L-1 TNF-α+10％blank serum),Duhuo Jisheng Decoction low(10 μg·L-1 TNF-α+2.5％drug-containing serum+7.5％blank serum),medium(10 pg·L-1 TNF-α+5％drug-con-taining serum+5％blank serum),high(10 μg·L-1 TNF-α+10％drug-containing serum)dose group.The concentration of serum containing Duhuo Jisheng Decoction was screened by MTT method.Cell prolifer-ation was detected using EdU staining.The expression of proliferation marker Ki67 was detected using immu-nofluorescence staining.The apoptosis rate was meas-ured by flow cytometry.The contents of IFN-γ,IL-4,IL-6 and IL-10 in each group were detected by ELISA.The mRNA and protein expression of Bax,Bcl-2,caspase-3 and TLR4/MyD88/NF-κB signaling pathway were detected by RT-qPCR and Western blot.Results Compared with the normal group,the cell prolifera-tion activity of the model group significantly increased,and the level of apoptosis decreased.The content of IFN-γ and IL-6 increased,and the content of IL-4 and IL-10 decreased.The TLR4/MyD88/NF-κB signaling pathway was activated.After the intervention of low,medium and high dose groups of serum containing Du-huo Jisheng Decoction,it could effectively improve the abnormal proliferation of cells and enhance apoptosis.At the same time,it inhibited the inflammatory re-sponse and the activation of TLR4/MyD88/NF-κB sig-naling pathway.Conclusions The serum containing Duhuo Jisheng Decoction can inhibit the abnormal pro-liferation of RA-FLS induced by TNF-α and the secre-tion of pro-inflammatory factors,and enhance apoptosis and anti-inflammatory levels.The mechanism may be related to the regulation of TLR4/MyD88/NF-κB sig-naling pathway.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective:To summarize the current research status, assessment tools, and influencing factors of fatigue in patients receiving maintenance hemodialysis (MHD) and provide a reference for the management of fatigue in this patient population.Methods:A literature search was conducted in databases including PubMed, Web of Science, ProQuest, Cochrane Library, Science Direct, Embase, CINAHL, China National Knowledge Infrastructure, WanFang Data, VIP, and China Biology Medicine disc for studies related to fatigue in patients receiving MHD. The search timeframe was from establishing the databases to January 23, 2024.Results:A total of 46 studies were included. Various assessment tools for fatigue in patients receiving MHD were identified, though specific tools were limited. The Short Form 36 Vitality Subscale (SF-36 VS) was the most commonly used assessment tool. The main factors influencing fatigue in these patients included sociodemographic, dialysis-related, disease-related, physical, nutritional, and psychological factors.Conclusions:Fatigue is a significant symptom in patients receiving MHD. Healthcare professionals must develop specific tools for accurately assessing fatigue in this population and explore standardized management plans.

Objective:To investigate the effect of feeder layer cells expressing interleukin(IL)-21 on the amplification of NK cells in vitro.Methods:The K562 cell line with IL-21 expression on its membrane was constructed by electroporation,and co-cultured with NK cells after inactivation.The proliferation of NK cells was observed.The killing function of the amplified NK cells in vitro was evaluated by the lactate dehydrogenase(LDH)and interferon-γ(IFN-y)release assay.A colorectal cancer xenograft model in NOD/SCID mice was established,and a blank control group,a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells in vivo.Results:K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation.After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days,the NK cells increased to 700 times,which showed an enhanced amplification ability compared with control group(P＜0.001).In the tumor cell killing experiment in vitro,there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells,and there was also no significant difference in mice in vivo.Conclusion:K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells in vitro,but do not affect the killing function of NK cells in vitro and in vivo.It can be used for the subsequent large-scale production of NK cells in vitro.

AIM To analyze the metabolites of nobiletin in rats in vivo based on characteristic ions.METHODS Ten rats were assigned into administration group and control group,and given intragastric administration of the 0.5％CMC-Na suspension of nobiletin(250 mg/kg)and 0.5％CMC-Na solution,respectively,after which plasma,urine and feces were collected,solid phase extraction method was adopted in pretreatment,UHPLC-HRMS analysis was performed.The candidate metabolites were systematically described according to diagnostic product ions,chromatographic retention time,accurate molecular weight and neutral loss fragments,after which accurate metabolites were obtained in the established metabolite data set with-CH3(m/z 15)characteristic ions as a baits.RESULTS A total of 64 metabolites were identified,whose main metabolic pathways were glucuronidation,sulfation,hydrogenation and their compound reactions.CONCLUSION This experiment elucidates the metabolites of nobiletin in rats in vivo,which provides a new reference for its further development.

AIM To optimize the extraction process for uracil,hypoxanthine,xanthine,uridine,thymine,inosine,guanosine and 2'-deoxyguanosine from Pheretima guillelmi(Michaelsen),and to determine their contents.METHODS With solid-liquid ratio,ultrasonic time and ultrasonic temperature as influencing factors,contents of hypoxanthine and total nucleosides as evaluation indices,the extraction process was optimized by orthogonal test.HPLC was adopted in the content determination of varioud nucleosides,the analysis was performed on a 30℃thermostatic Agilent C18 column(4.6 mm×250 mm,5 μm),with the mobile phase comprising of methanol-water flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS The optimal conditions were determined to be 1∶250 for solid-liquid ratio,60 min for ultrasonic time,and 60℃for ultrasonic temperature.Eight nucleosides showed good linear relationships within their own ranges(R2＞0.999 0),whose average recoveries were 99.11%-103.27%with the RSDs of 0.85%-2.89%.CONCLUSION This stable and reliable method can be used for the extraction and content determination of nucleosides from P.guillelmi.

Objective:To investigate the correlation between the variability of blood uric acid level and the progression of type 2 diabetic nephropathy and retinopathy.Methods:A total of 240 patients with type 2 diabetic nephropathy were selected from a cohort established in Pinggu District Hospital of Beijing in 2015 for retrospective analysis. The blood uric acid level of the patients was measured, the variability of uric acid level was calculated, and the patients were divided into group A, group B, group C and group D according to the quartile of uric acid variability, with 60 cases in each group. The subjects were followed up, and their general information, biochemical indicators, diabetic nephropathy and diabetic retinopathy were collected. According to the diabetic nephropathy and retinopathy during follow-up, the subjects were divided into progressive group and non-progressive group, so as to further clarify the correlation between the variability of blood uric acid level and the progression of diabetic nephropathy and retinopathy.Results:Up to the last follow-up date in July 2022, a total of 24 cases were lost to follow-up in group A, 27 cases in group B, 20 cases in group C, and 22 cases in group D. Finally, 36 cases were included in group A, 33 cases in group B, 40 cases in group C, and 38 cases in group D. There was no significant difference in age, gender, body mass index, systolic blood pressure, diastolic blood pressure, cholesterol, triglyceride, low density lipoprotein cholesterol, fasting blood glucose, glycated hemoglobin and serum creatinine among four groups ( P>0.05). Univariate analysis showed that the incidence of diabetic nephropathy and retinopathy progression increased with the increase of the quartile of uric acid variability in patients with type 2 diabetes (the incidences of progression in A, B, C and D groups were 16%, 49%, 63% and 79%, F = 0.95, P<0.05). Pearson correlation analysis showed that blood uric acid variability was positively correlated with the progression of diabetic nephropathy and retinopathy ( r = 0.482 and 0.501, P<0.05). Logistics regression analysis showed that with the increase of the quartile of uric acid variability, the progression risk of diabetic nephropathy ( OR = 3.521, 5.226 and 6.548; P<0.05) and retinopathy ( OR = 3.733, 4.844 and 5.872; P<0.05) in type 2 diabetes patients increased gradually. Conclusions:The variability of blood uric acid level is positively correlated with the progression of type 2 diabetic nephropathy and retinopathy. The higher the risk of progression of diabetic nephropathy and retinopathy with the increase of quartile of blood uric acid level variability, the more important it is to regularly monitor the blood uric acid level of type 2 diabetic patients.

Background and purpose:Heterogeneous nuclear ribonucleoprotein K(hnRNPK)is an RNA special binding protein that participates in regulating the expression of related genes and protein translation.It has been linked to the malignant occurrence and development of various tumors,but its role in breast cancer remains unclear.The aim of this study was to investigate the effects of hnRNPK on ferroptosis in breast cancer cells and the underlying mechanisms.Methods:Based on The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases,hnRNPK expression in breast cancer tissues and normal tissues and its relationship with clinical prognosis were analyzed by bioinformatics.We detected hnRNPK expression in breast cancer cells and tissues using real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR),Western blot,and immunohistochemistry staining diagnosis methods.MCF-7 and MDA-MB-231 breast cancer cells were transfected with siRNA,and divided into control group(control),empty body group(NC),and interference vector group(si-hnRNPK).Cell proliferation was detected by cell counting kit-8(CCK-8)and plate clone formation assays.RNA-seq analysis was applied to explore potential targeted biological functions and signaling pathways affected by hnRNPK.Additionally,we investigated the impact of hnRNPK on ferroptosis phenotype using Western blot and commercial kits for reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH),and Fe2+,ferroptosis inhibitor(ferrostatin-1,Fer-1)was used to detect the rescue effect of hnRNPK knockdown on ferroptosis.The impact of hnRNPK on the expressions of Wnt/β-catenin pathway-related proteins were determined by Western blot.Results:The bioinformatics analyses indicated hnRNPK was upregulated in breast cancer tissues(P＜0.01),and the overall survival of patients in the high expression group was poorer compared with those in the low expression group(P＜0.05).hnRNPK was highly expressed in breast cancer tissues and cells,and knocking down hnRNPK weakened the proliferation ability of breast cancer cells(P＜0.05).The RNA-seq analysis showed that hnRNPK was significantly enriched in ferroptosis,apoptosis,and the Wnt/β-catenin signaling pathway.Knocking down hnRNPK promoted ferroptosis in breast cancer cells by inducing lipid ROS and MDA,as well as Fe2+accumulation(P＜0.05).Interestingly,the ferroptosis inhibitor ferrostatin-1(Fer-1)reversed the promotive effect of hnRNPK knockdown on ferroptosis(P＜0.05).Downregulation of hnRNPK led to a decrease in the expressions of β-catenin and c-Myc in the Wnt/β-catenin signaling pathway,while expressions of CK1α,APC and the GSK-3β complex were elevated(P＜0.05).Conclusion:hnRNPK is highly expressed in breast cancer,and knocking down hnRNPK promotes ferroptosis in breast cancer cells by inhibiting the Wnt/β-catenin signaling pathway,thereby suppressing the malignant progression of breast cancer.

Diabetes cardiomyopathy(DCM)is a cardiovascular disease with structural damage and dysfunction of myocardium under the condition of glucose and lipid metabolism disorder in diabetes.The main pathological characteristics are ventricular hypertrophy,myocardial remodeling and cardiomyocyte death,which induce heart failure.DCM has become one of the main causes of death in patients with diabetes.As a pro-inflammatory programmed cell death pathway,pyroptosis mediated by inflammasomes is considered an important driver of chronic inflammation due to its exces-sive activation.Recently,it was found that the over activation of pyroptosis promoted the course of DCM and became the hub connecting the disorder of glucose and lipid metabolism in diabetes and myocardial injury.With the discovery of more and more roles of Traditional Chinese Medicine(TCM)in regulating pyroptosis,TCM affects the occurrence and develop-ment of DCM by regulating pyroptosis or protein expression,which has become a research hotspot of myocardial protection strategies in diabetes.This article reviews the mechanism of pyroptosis in DCM and the research progress of TCM mono-mers and compound prescriptions in regulating the process of pyroptosis in DCM,in order to provide new ideas and targets for clinical research and treatment of DCM.