Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

The prevention and control of emerging and reemerging infectious diseases are crucial for national biosecurity, and surveillance and reporting of pneumonia of unknown etiology are main ways for the early detection of these diseases and mitigation of their severity. This paper summaries the surveillance methods for pneumonia of unknown etiology and emerging and reemerging infectious diseases globally, indicating that such surveillance is mainly conducted based on hospital. Western countries primarily combine active and passive surveillance while utilizing artificial intelligence technology to rapidly identify cases. China mainly use passive surveillance based on the surveillance system for pneumonia of unknown etiology, with the function of early warning in the identification of emerging infectious diseases, such as avian influenza. However, with the improvement in the surveillance system operation, the overlap with other surveillance disease systems, such as influenza, has occurred, and the improvements in case definition and operational protocol are needed. It is recommended to improve the specificity of the case definition, strenthen training in hospital staff, inclduing clinical workers and office workers, and formulate incentive mechanisms. It is necessary to emphasize the responsibility of clinicians as the main force for the detection and reporting of pneumonia of unknown etiology and emerging infectious diseases, improve the appilication of artifical intelligent technique and conduct multi-source surveillance, such as third-party testing.

Objective:To compare the burden of benign prostatic hyperplasia（BPH）between China and the United States from 1990 to 2021.Methods:The prevalence，incidence，years lived with disability（YLD），and their age-standardized rates for BPH in China and the United States from 1990 to 2021 were obtained from the Global Burden of Disease Study 2021（GBD 2021）. The average annual percentage change（AAPC）of the age-standardized incidence rate（ASIR）and the age-standardized YLD rate（ASYR）was calculated using Joinpoint regression analysis. In addition，the YLD burden of BPH，prostate cancer，kidney cancer，bladder cancer，and three other urological diseases were compared between the two countries.Results:From 1990 to 2021，the number of BPH cases in China increased from 1.460 4 million to 3.244 5 million，the number of prevalent cases rose from 9.940 5 million to 23.111 2 million，and YLDs grew from 0.2 million person-years to 0.460 2 million person-years，with AAPCs of 2.63%，2.78%，and 2.75%，respectively. In 2021，the numbers of incident cases，prevalent cases，and YLDs were 0.577 9 million，4.930 3 million，and 0.095 9 million person-years in the United States，and 13.787 6 million，112.502 million，and 2.235 7 million person-years globally. China’s ASIR decreased from 363.07/100 000 to 299.14/100 000（AAPC -0.60%），and ASYR from 57.33/100 000 to 45.84/100 000（AAPC -0.70%），both of which were higher than those in the United States but lower than the global level. Age-specific analyses showed declining incidence and YLD rates across all age groups in China，while certain age groups in the United States demonstrated increasing trends. From 1990 to 2021，the proportion of YLDs attributable to BPH among seven urological diseases in China rose from 61.4% to 69.2%. In 2021，YLDs due to prostate cancer accounted for the highest proportion among seven urinary system diseases in the United States，reaching 54.5%. Projections indicate that although ASIR and ASYR in China will decline from 2022 to 2040，the absolute numbers of incident cases and YLDs are projected to continue to rise，reaching 4.97 million and 0.78 million，respectively，by 2040.Conclusions:Between 1990 and 2021，the number of incidence cases，prevalence cases，and YLDs of BPH in China increased markedly，while ASIR and ASYR declined. The disease burden of BPH remains substantial，with a higher proportion of YLDs among urological diseases compared with the United States. By 2040，the number of BPH cases and YLDs in China is projected to further increase，underscoring the need for greater public health attention.

AIM To investigate the tumor-suppressive mechanism of Guiqi Yiyuan Extract combined with cisplatin in Lewis lung cancer mice.METHODS Ten intact C57BL/6J mice were assigned to the blank group.Sixty additional mice were developed into Lewis lung cancer models bearing transplanted tumor and subsequently allocated into the model group,the cisplatin group(5 mg/kg),the high-dose Guiqi Yiyuan Extract group(6.6 g/kg),and the low-dose,medium-dose and high-dose Guiqi Yiyuan Extract combined with cisplatin group(1.6,3.3,6.6 g/kg+5 mg/kg),with 10 mice in each group.Mice in the blank and model groups received saline via daily gavage,while treatment groups were administered Guiqi Yiyuan Extract orally(once daily),and cisplatin injection intraperitoneally(once every other day).After 14 days of drug administration,mice were euthanized for endpoint analysis.The following assessments were conducted:general health status and body weight changes monitored throughout the study period;tumor excision and weighing for inhibition rate calculation;histopathological examination of tumors via hematoxylin-eosin(HE)staining;serum quantification of IL-1 β,IL-18 and HMGB1 by ELISA;ultrastructural analysis of tumor cell death using transmission electron microscopy(TEM);spatial localization of TXNIP and GSDMD-N in tumor sections via immunofluorescence(IF);and Western blot detection of TXNIP,NLRP3,Caspase-1,cleaved Caspase-1,GSDMD,GSDMD-N protein expressions in tumor tissues.RESULTS Compared to the model group,the cisplatin group and all combination therapy groups exhibited significant reduction in tumor weight(P＜0.05)and increased tumor suppression rate;enhanced tumor tissue necrosis with characteristic pyroptotic morphology;elevated serum levels of IL-1β,IL-18 and HMGB1(P＜0.05);and upregulated expressions of pyroptosis-associated proteins TXNIP,NLRP3,Caspase-1,cleaved Caspase-1,GSDMD and GSDMD-N(P＜0.05).The high dose combination group demonstrated optimal therapeutic efficacy(P＜0.05).CONCLUSION Guiqi Yiyuan Extract enhances cisplatin sensitivity,demonstrating synergistic anti-tumor effects in Lewis lung carcinoma-bearing mice.This combinatorial therapeutic effect likely involves modulation of the TXNIP/NLRP3/Caspase-1/GSDMD pathway.

AIM To investigate the effects of Congrong San on neuronal apoptosis and the Bax/Bcl-2/Caspase3 signaling pathway in a rat model of Alzheimer's disease(AD).METHODS A total of 60 2-month-old SD male rats were randomly divided into the blank group,the model group,the memantine hydrochloride group(0.025 g/kg)and low-dose,medium-dose and high-dose Congrong San groups(4.62,9.24,18.48 g/kg).All groups except the control group received stereotactic intracerebral injection of Aβ1-42 to establish AD models.Following the successful modeling,each group received its corresponding intragastric administration once daily for 28 consecutive days.After the administration,the rats had their learning and memory ability detected by the morris water maze test;their hippocampal neuronal morphology observed with HE and Nissl staining;their hippocampal neuronal apoptosis observed with TUNEL staining;and their hippocampal expressions of amyloid precursor protein(APP),β-site APP-cleaving enzyme 1(BACE1),and apoptosis-related proteins Bax,Bcl-2 and Caspase3 detected with Western blot assay.RESULTS Compared with the model group,the groups intervened with medium-dose and high-dose Congrong San exhibited improved learning and memory performance,alleviated hippocampal neuronal damage,increased Nissl body count(P＜0.01),reduced hippocampal apoptosis rate(P＜0.05,P＜0.01),decreased protein expressions of APP,BACE1,Bax and cleaved-Caspase3/Caspase3 ratio(P＜0.05,P＜0.01),and elevated Bcl-2 expression(P＜0.01).CONCLUSION Congrong San mitigates cognitive impairment,hippocampal neuronal damage,and apoptosis in AD rats,probably through inhibition of the Bax/Bcl-2/Caspase3 signaling pathway activation.

Objective To explore the predictive value of dose surface histogram(DSH)in image guided radiotherapy(IGRT)for radiotherapy-induced acute radiation proctitis(ARP)in cervical cancer(CCA).Methods Totally 380 patients with CCA IGRT admitted to some hospital from May 2019 to May 2023 were selected prospectively and randomly divided into a control group(n=1 80)and an experimental group(n=200).The patients in the 2 groups were followed up and the incidence rates of ARP were counted,and rectal dose distribution was evaluated using dose volume histogram(DVH)in the control group and DSH in the experimental group.The predictive values of DVH and DSH for ARP were evaluated and compared using ROC curves.Statistical analysis was performed using SPSS 21.0 software.Results The two groups did not have statistically significant difference in the incidence rate of ARP(P＞0.05),while there were significant differences in the evaluation indicators of the rectal dose distribution(P＜0.05).V40,V50,S40 and S50 proved to have low predictive values for grade Ⅰ-Ⅳ ARP with AUC 0.700(P＜0.05);V60 and S60 had moderate predictive values for grade Ⅰ-Ⅳ ARP with AUC greater than 0.700 and less than or equal to 0.900(P＜0.05);V70,V78,S70 and S7s showed high predictive values for grade Ⅰ-Ⅳ ARP with AUC higher than 0.900(P＜0.05).Delong's test results indicated that DVH and DSH had no significant differences in AUC when used to predict gradeⅠ-Ⅳ ARP(allP＞0.05).Conclusion DSH is essentially the same as DVH when used for the prediction of grade Ⅰ-Ⅳ ARP due to CCA IGRT,and thus can be used for the supplementation and optimization of radiotherapy planning systems.[Chinese Medical Equipment Journal,2025,46(3):48-53]

This work was aimed at analyzing the protein characteristics of Coiled-Coil Domain-Containing Protein 115(CCDC115)and using Ccdc115-deficient mouse monocyte-macrophage leukemia cells(RAW264.7)to explore the influence of CCDC115 on the intracellular survival of Salmonella Typhimurium.Bioinformatics analysis was conducted to examine the fundamental attributes of CCDC115,which was determined to be an unstable protein consisting of two α-helices and an intervening disordered re-gion,devoid of any transmembrane structural domains.A RAW264.7-Ccdc115-KO cell line was successfully established with CRISPR/Cas9 gene-editing technology.To elucidate the effects of CCDC115 on the intracellular survival of Salmonella Typhimurium,we infected RAW264.7 cells with Salmonella Typhimurium.The expression of CCDC115 was found to be upregulated at both the mRNA and protein levels post-infection,according to RT-qPCR and western blot analysis.Via counting of colony-forming units(CFU),the proliferation rate of Salmonella Typhimurium within RAW264.7-Ccdc115-KO cells was found to be 1.5-fold higher than that in RAW264.7 cells.Acidification imaging studies indicated that,whereas Salmonella Typhimurium phagosomes underwent acidifi-cation in RAW264.7 cells,this process was absent in RAW264.7-Ccdc115-KO cells.In conclusion,the study successfully estab-lished a RAW264.7-Ccdc115-KO cell line and demonstrated that the expression of CCDC115 is elevated during Salmonella Ty-phimurium infection,thus potentially inhibiting the intracellular survival of Salmonella Typhimurium by facilitating phagosome acidifi-cation.This study lay a theoretical foundation for functional studies of CCDC115 and the investigation of mechanisms regulating the survival of intracellular Salmonella Typhimurium.

Cardiovascular diseases that develop from hypercholesterolemia-induced atherosclerosis have emerged as a significant threat to human health.Recently,probiotics exhibiting cholesterol-lowering properties have emerged as a prominent area of research.Numerous studies have demonstrated that Lactobacillus reuteri can effectively reduce endogenous cholesterol synthesis,regulate cholesterol transport,and promote cholesterol degradation by modulating the expression of key genes,such as sterol-regulatory element binding protein 2,3-hydroxy-3-methyl-glutaryl coenzyme A reductase,and cholesterol 7 alpha-hydroxylase,in both the liver and intestinal epithelial cells of the host.This leads to a notable decrease in total cholesterol and low-density lipoprotein cholesterol levels in the host serum.The present paper offers a comprehensive overview of the underlying mechanisms responsible for the cholesterol-lowering effects exerted by L.reuteri,aiming to provide valuable insights into the treatment of hypercholesterolemia and the development of probiotics with cholesterol-lowering properties.

Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P＜0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.