OBJECTIVE To explore the intervention mechanism of Bushen anzhi decoction （BSAZD） in rats with insomnia induced by kidney failing to store spirit. METHODS A total of 36 SD rats were randomly divided into normal group （normal saline）， model group （normal saline）， eszopiclone group （positive control group， 0.27 mg/kg）， and low-， medium- and high-dose BSAZD groups （5.85， 11.70， 23.40 g/kg）， with 6 rats in each group. Except for the normal group， rats in the remaining groups were subjected to combined administration of D-galactose and DL-4-chlorophenylalanine to establish the rat model of insomnia due to kidney failing to store spirit. After successful modeling， corresponding liquid medicine or normal saline was intragastrically administered once daily for 14 consecutive days. After the last administration， behavioral indicators and hippocampal histopathological morphology of rats were detected. Untargeted metabolomics was adopted to screen serum differential metabolites and enrich relevant signaling pathways. RESULTS Compared with the model group， the high-dose BSAZD group exhibited significantly increased open arm entries ， open arm time and sleep time （ P ＜0.05）， accompanied by markedly decreased closed arm entries， closed arm time and sleep latency （ P ＜0.05）. Hippocampal neurons in the high-dose BSAZD group were arranged regularly， and the size， morphology and location of cell nuclei were nearly normal. Untargeted metabolomics analysis identified oxidized glutathione， glutamate and other differential metabolites in serum of rats in the high-dose BSAZD group； these differential metabolites were mainly enriched in GABAergic synapse， glutamatergic synapse， cAMP signaling pathway and 2-oxocarboxylic acid metabolism pathway. CONCLUSIONS BSAZD can improve sleep quality， relieve anxiety-like behaviors in rats with insomnia due to kidney failing to store spirit. Its therapeutic mechanism may be associated with regulating GABAergic and glutamatergic synaptic pathways to restore neurotransmitter balance and further alleviate oxidative stress injury， modulating the cAMP signaling pathway to facilitate neural repair， and intervening the 2-oxocarboxylic acid metabolism pathway to optimize energy supply.

OBJECTIVE To explore the intervention mechanism of Bushen anzhi decoction （BSAZD） in rats with insomnia induced by kidney failing to store spirit. METHODS A total of 36 SD rats were randomly divided into normal group （normal saline）， model group （normal saline）， eszopiclone group （positive control group， 0.27 mg/kg）， and low-， medium- and high-dose BSAZD groups （5.85， 11.70， 23.40 g/kg）， with 6 rats in each group. Except for the normal group， rats in the remaining groups were subjected to combined administration of D-galactose and DL-4-chlorophenylalanine to establish the rat model of insomnia due to kidney failing to store spirit. After successful modeling， corresponding liquid medicine or normal saline was intragastrically administered once daily for 14 consecutive days. After the last administration， behavioral indicators and hippocampal histopathological morphology of rats were detected. Untargeted metabolomics was adopted to screen serum differential metabolites and enrich relevant signaling pathways. RESULTS Compared with the model group， the high-dose BSAZD group exhibited significantly increased open arm entries ， open arm time and sleep time （ P ＜0.05）， accompanied by markedly decreased closed arm entries， closed arm time and sleep latency （ P ＜0.05）. Hippocampal neurons in the high-dose BSAZD group were arranged regularly， and the size， morphology and location of cell nuclei were nearly normal. Untargeted metabolomics analysis identified oxidized glutathione， glutamate and other differential metabolites in serum of rats in the high-dose BSAZD group； these differential metabolites were mainly enriched in GABAergic synapse， glutamatergic synapse， cAMP signaling pathway and 2-oxocarboxylic acid metabolism pathway. CONCLUSIONS BSAZD can improve sleep quality， relieve anxiety-like behaviors in rats with insomnia due to kidney failing to store spirit. Its therapeutic mechanism may be associated with regulating GABAergic and glutamatergic synaptic pathways to restore neurotransmitter balance and further alleviate oxidative stress injury， modulating the cAMP signaling pathway to facilitate neural repair， and intervening the 2-oxocarboxylic acid metabolism pathway to optimize energy supply.

Objective: To analyze the serological phenotypes and molecular genetic characteristics of a proband with ABO forward and reverse typing discrepancy and their family members, and to investigate the molecular genetic mechanism underlying the Ael/AelB phenotype caused by the c. 575T>C variant on an ABO A1.02 background. Methods: Serological testing and adsorption-elution assays were performed for blood group identification of the proband and family members. PCR-SBT, cloning sequencing, and Nanopore third-generation sequencing were used to analyze ABO gene variants and haplotypes. Phyre2 and PyMOL were used to predict the effect of the p. Ile192Thr variant on the structure of glycosyltransferase A (GTA), and plasma GTA activity was evaluated using an in vitro transferase assay. Results: Among thirteen family members, four members showed a serological phenotype consistent with the Ael subtype and one with the AelB subtype. Cloning sequencing and third-generation sequencing confirmed that five family members carried the ABO A1.02 (c. 575T>C) variant, which co-segregated with the weak A phenotype in the family. Structural modeling suggested that p. Ile192Thr may alter the local spatial conformation and hydrogen-bond interactions of GTA. Plasma GTA activity testing showed markedly reduced GTA activity in the tested individuals. Conclusion: The ABO A1.02 (c. 575T>C) variant is associated with the Ael/AelB phenotype. The p. Ile192Thr substitution may weaken A antigen expression by altering GTA conformation and reducing enzymatic activity. This study provides additional evidence for the role of this variant in the Ael/AelB phenotype from the perspectives of familial inheritance, full-length haplotype analysis, and functional assessment, which is important for accurate ABO subtype identification and individualized transfusion strategies.

OBJECTIVE To investigate the cognition， attitudes， and current practices of Chinese physicians regarding biosimilars and their clinical switching， so as to provide a basis for the rational clinical application of biosimilars in China and the improvement of related policies.METHODS A national cross-sectional online questionnaire survey was conducted in 2025. Quota sampling was adopted， with quotas based on physicians’ sex， city tier， and specialty. Questionnaire invitations were distributed to licensed physicians， and physicians who had prescribed monoclonal antibody biosimilars were included for analysis. The questionnaire covered their familiarity with biosimilars， attitudes， clinical switching practices， and reasons. Data were processed using reliability and validity tests， dispersion analysis， and cross-tabulation analysis.RESULTS The questionnaire scale showed good reliability and validity， with Cronbach’s α coefficient ＞0.7 and KMO value ＞0.6. A total of 2 071 survey invitations were distributed， and 300 valid questionnaires were returned， with an effective response rate of 14.5%. Among the respondents， 75.3% had prescribed 3-5 types of monoclonal antibody biosimilars； only 35.3% were aware that China currently has no clear regulatory provisions on biosimilar switching； 46.3% and 47.3% of respondents believed that there was no difference in safety and efficacy， respectively， between biosimilars and originator products； 63.0% of respondents were concerned about the immunogenicity risk of long-term switching between originator drugs and biosimilars. A total of 92.3% of respondents had experience with biosimilar switching， 87.7% of which involved switching from originator products to biosimilars， and 28.2% of respondents had completed three or more switches in the same patient. Among the primary reasons for switching， non-medical factors accounted for 57.4%， higher than medical factors at 42.6%； “lower treatment cost after switching” was the most important reason for switching； 95.3% of respondents believed that informed consent should be obtained from patients before switching.CONCLUSIONS The surveyed physicians had basic knowledge of biosimilars， but their understanding of the relevant regulations of the national drug regulatory authority was insufficient. Although most physicians were concerned about the risks of non-medical switching， clinical switching was mostly driven by non-medical factors. It is recommended to strengthen policy education for medical professionals and standardize clinical switching behavior. The National Medical Products Administration should also promptly improve the interchangeability review standards and post-marketing surveillance system for biosimilars, so as to promote the rational use of biosimilars.

OBJECTIVE To evaluate the clinical efficacy and safety of eravacycline in intensive care unit （ICU） patients with carbapenem-resistant Acinetobacter baumannii （CRAB） and/or carbapenem-resistant Klebsiella pneumoniae （CRKP） pneumonia.METHODS Clinical data of patients with CRAB and/or CRKP pneumonia who were admitted to the ICU of Peking Union Medical College Hospital， Chinese Academy of Medical Sciences， and received eravacycline treatment from July 2023 to November 2024 were collected. The 28-day all-cause mortality rate， clinical failure rate， 28-day ICU-free days among surviving patients， 28-day mechanical ventilation-free days among surviving patients， microbiological clearance， and adverse reactions were recorded.RESULTS A total of 37 patients were included. Among the 37 patients， 11 （29.73%） experienced clinical failure， and the 28-day all-cause mortality rate was 18.92% （7/37）. The 28-day ICU-free days among surviving patients were （14.60±10.61） days， and the 28-day mechanical ventilation-free days among surviving patients were （24.55±4.29） days. Microbiological clearance was achieved in 12 patients （32.43%）， and the mean time to conversion was （10.58±12.05） days. Adverse reactions occurred in 5 patients （13.51%）， including 2 cases of elevated liver enzymes， 2 cases of coagulopathy， and 1 case of drug hypersensitivity syndrome.CONCLUSIONS Eravacycline can effectively control systemic bacterial infection and inflammatory response in ICU patients with pneumonia caused by CRAB and/or CRKP. However， patients with underlying liver disease or receiving multiple antibacterial agents should be alert to elevated liver enzymes， coagulation abnormalities， and the rare drug hypersensitivity syndrome.

ObjectiveTo explore the process and guidelines for ethical review in decentralized drug clinical trials， promote clinical trial progress， and ensure drug development progress. MethodsThe key points of the ethical review were summarized by studying the relevant laws and regulations on decentralized drug clinical trials， analyzing the advantages and challenges of decentralized drug clinical trials， and combining the experience of the ethics committee of the institution in reviewing decentralized drug clinical trials. ResultsRelevant laws and regulations were the basis for the ethical review， and the ethics committee should adopt appropriate review methods based on regulations and hospital ethical standard operating procedures. The ethics committee should focus on the feasibility， applicability， and rationality， the adequacy of informed consent， the protection of rights and interests and privacy of subjects， as well as the qualification and standard operating procedures of electronic platforms for conducting decentralized drug clinical trials. ConclusionDecentralized drug clinical trials are in their early stages and urgently require guidance from relevant laws and regulations. Ethical review is also constantly being refined through exploration. It is necessary to supervise the implementation of responsibilities by all parties， pay attention to the rights and interests of subjects， and gradually promote the implementation of decentralized drug clinical trials.

Objective : To explore the correlation between uteroglobulin-related protein 1(UGRP1) and primary hypothyroidism. Methods : Ninety-six patients with primary hypothyroidism were selected, including 66 patients with positive thyroid peroxidase antibodies(TPOAb) or anti-thyroglobulin antibodies(ATG) as the antibody-positive group, 30 patients with negative thyroid autoantibodies as the antibody-negative group, and 96 healthy people as the control group. The general clinical data, thyroid-related indicators and serum UGRP1 levels were compared among these three groups. Human thyroid normal cells(NTHY-ORI 3-1) were transfected with plasmids in vitro, thus establishing the control group as well as the UGRP1 group. Enzyme linked immunosorbent assay(ELISA) was used to detect and compare the T4 level in the cell culture supernatant. Results : The differences in thyroid stimulating hormone(TSH), TPOAb and ATG among the three groups were statistically significant(P<0.05). The serum UGRP1 levels in the antibody-positive(303.97±156.00) pg/ml and antibody-negative groups(352.13±188. 37) pg/ml were higher than those in the control group( 237. 54 ± 137. 20) pg/ml,and the differences between the groups were statistically significant( P = 0. 005). Meanwhile,there was no statistically significant difference between the antibody-positive and antibody-negative groups. Multi-factor Logistic regression analysis showed that UGRP1 was the risk factor for the occurrence of primary hypothyroidism( OR = 1. 004,95% CI: 1. 001-1. 007,P =0. 007). The difference between the control group and UGRP1 group in T4 concentration secreted by human thyroid normal cells was not statistically significant. Conclusion Serum UGRP1 levels increase in patients with primary hypothyroidism,and the high expression of UGRP1 may have no direct relation to the function of thyroid cells secreting T4.

Objective To explore the expression level of plasma miRNA in patients with swallowing and cognitive dysfunction after ischemic stroke and its correlation with the severity of dysfunction.Methods A retrospective analysis was conducted on the clinical data of 109 patients with ischemic stroke who were hospitalized in the First Affiliated Hospital of Guangxi Medical University and the First Affiliated Hos-pital of Fujian Medical University from January 2023 to March 2024.The expression levels of plasma miR-140-5p,miR-17-5p,and miR-103a-3p were detected by RT-qPCR.The swallowing function was evaluated by the Wada Drinking Water Test and the Functional Oral Feeding Scale,and the cognitive function of stroke pa-tients was evaluated by the modified Rankin Scale(mRS)classification.According to the mRS rating criteria,the patients were divided into the good functional outcome group(grade≤2,n=50)and the poor functional outcome group(grade≥3,n=59).Analyzed the correlation between miRNA expression levels and the degree of functional impairment,compared the clinical characteristics of patients with different mRS grades,and used the receiver operating characteristic(ROC)curve and the area under the curve(AUC)to analyze the efficacy of miRNA in predicting the functional outcome of mRS evaluation.Results The results of Spearman correla-tion analysis showed that the expression level of miR-17-5p was negatively correlated with the classification of the Wada Drinking Water Test(r=-0.317)and the classification of the Functional Oral Feeding Scale(r=-0.457,P<0.05).Compared with the good functional outcome group,the poor functional outcome group had a lower proportion of males,shorter disease course,and higher expression level of miR-103a-3p,the differ-ences were statistically significant(P<0.05).The results of ROC curve analysis showed that miR-103a-3p(AUC=0.709,95%CI:0.611-0.808)had the ability to distinguish functional outcomes in mRS evaluation.However,miR-140-5p(AUC=0.514,95%CI:0.405-0.624)and miR-17-5p(AUC=0.527,95%CI:0.414-0.637)did not have the ability.Conclusion The expression level of plasma miR-17-5p is related to the severi-ty of dysphagia in patients in the recovery period of ischemic stroke,and miR-103a-3p is helpful for judging the functional outcomes of patients in the recovery period of ischemic stroke.

Objective To optimize the platelet enrichment method,and to analyze the concentration changes of key molecules in platelet-rich plasma(PRP)before and after activation,as well as the impact of its activated products on the proliferation of rat endothelial progenitor cells.Methods The tube double-centrifu-gation method was employed to optimize platelet enrichment,and the platelet count in the enriched PRP was measured.ELISA was used to detect the concentration changes of vascular endothelial growth factor(VEGF),endostatin(ES),and P-selectin(CD62P)in PRP before and after activation.The PRP was activated by using liquid nitrogen freeze-thaw method,and the effect of its activated products on the proliferation of rat endothelial progenitor cells was evaluated by using the methyl thiazolyl tetrazolium(MTT)assay.Results The optimal enrichment coefficient of platelets achieved by the double-centrifugation method was 4.63.After low-speed,long-duration double centrifugation,the platelet count was highest in the upper layer of the buffy coat.For PRP with a platelet count of 500× 109/L obtained by machine collection,the VEGF con-centrations before and after activation were(3 418.12±488.80)pg/mL and(4 530.04±308.30)pg/mL,re-spectively,the ES concentrations were(6 168.98±253.22)pg/mL and(6 594.65±82.47)pg/mL,respec-tively,the CD62P concentrations were(6 678.23±324.15)pg/mL and(17 630.53±746.24)pg/mL,respec-tively,statistically significant differences were observed in the above indicators before and after activation(P＜0.01).The activated PRP was diluted in a gradient manner by using a specialized culture medium for en-dothelial progenitor cells.MTT assay results indicated that,in the basal medium,the optimal volume fraction for promoting endothelial progenitor cell proliferation was 0.25％after 48 hours of culture;in the complete medium,the optimal volume fractions for promoting endothelial progenitor cell proliferation were 0.062 5％after 24 hours and 0.125％after 48 hours.Conclusion The concentrations of VEGF,ES,and CD62P in the optimized,enriched PRP exhibited significant changes before and after activation.The optimal volume fraction for promoting endothelial progenitor cell proliferation in the basal medium was 0.25％.

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.