AIM: To investigate and analyze serum nitric oxide synthase 4(NOX4)and matrix metalloproteinase(MMP)-2 expressions in patients with diabetes mellitus(DM)complicated with neovascular glaucoma(NVG)and their clinical significance.METHODS: A prospective study was conducted on 161 patients with DM complicated with NVG admitted to Handan City Eye Hospital(The Third Hospital of Handan)from June 2020 to June 2023. Based on whether complications occurred 1 a after trabeculectomy in the study group patients, they were divided into a group with good prognosis(n=90)and a group with poor prognosis(n=71). During the same period, 161 patients with chronic angle-closure glaucoma without iris neovascularization were selected as the control group. ELISA method was applied to detect the expression levels of serum NOX4 and MMP-2. ROC curve was plotted to evaluate the diagnostic value and postoperative complication prediction value of NOX4 and MMP-2 in patients with DM complicated with NVG. Pearson method was applied to analyze the correlation of NOX4 and MMP-2 with vascular endothelial growth factor(VEGF)and interleukin-6(IL-6). Multivariate Logistic regression was applied to analyze the influencing factors of postoperative complications in the study group.RESULTS:The general information of the study group and control group of patients is comparable. Compared with the control group, the expression levels of serum NOX4, MMP-2, VEGF, and IL-6 in the study group were significantly increased(P<0.001). According to Pearson analysis, serum NOX4 and MMP-2 levels significantly positively correlated with VEGF and IL-6 levels, respectively(P<0.001). According to ROC curve, the AUC of NOX4 combined with MMP-2 in the diagnosis of DM complicated with NVG was better than that of individual diagnosis of NOX4(Z=3.341, P<0.05)and MMP-2(Z=2.788, P<0.05). The duration of DM, the proportion of people with intraocular pressure >21 mmHg, and the expression levels of NOX4, MMP-2, VEGF, and IL-6 in the poor prognosis group were all higher than those in the good prognosis group(P<0.001). The duration of DM, intraocular pressure >21 mmHg, the levels of NOX4, MMP-2, VEGF, and IL-6 were all risk factors for poor prognosis in DM patients with NVG(P<0.001). According to ROC curve, the combined prediction of NOX4 and MMP-2 for postoperative complications in patients with DM complicated by NVG was superior to the AUC predicted by NOX4(Z=3.727, P<0.05)and MMP-2(Z=2.219, P<0.05), respectively.CONCLUSION:NOX4 and MMP-2 are upregulated in the serum of patients with DM complicated by NVG. Combined detection of these two markers holds significant clinical value for the early diagnosis and prognosis of patients with DM complicated by NVG.

AIM: To detect the distribution and expression of vascular endothelial growth factor(VEGF)in radiation retinopathy(RR)through fluorescence targeted imaging.METHODS:Covalent binding of fluorescein FITC with VEGF antibody ranibizumab to prepare targeted fluorescent imaging probe ranibizumab-FITC. SD rats were randomly divided into three groups based on the principle of weight balance: a normal control group(Con group), a low-dose radiation group(10 Gy group), and a high-dose radiation group(30 Gy group). Medical linear accelerators and lead blocks were used to locally irradiate the rat eyeballs for modeling. Western blot and qRT-PCR were used to detect the expression levels of VEGF-A in each group and to screen for appropriate modeling dose. The inverted fluorescence microscope and the confocal microscope were used to observe the distribution of VEGF and imaging probes in the retinas of control and RR model group rats, and to verify the effectiveness of targeted probes.RESULTS:The expression level of VEGF-A in the retina of rats in the high-dose radiation group(30 Gy group)was higher than that in the normal control group(Con group). In early RR, VEGF expression was observed to be associated with microaneurysms and abnormal microvessels in the retina. VEGF accumulation was observed at the site of capillary wall damage. When retinal capillary endothelial damage occurred, targeted probes gathered on the outer surface of the vessel wall.CONCLUSION:The expression level of VEGF in the retina of RR model rats is elevated, and fluorescent targeted molecular imaging probes can detect the spatial distribution of VEGF at the microvascular lesions in the retina of RR rats.

AIM: To investigate the relationship between serum levels of sirtuins(Sirts)1-6 and oxidative stress markers, and to evaluate their diagnostic value for age-related cataract(ARC).METHODS:A prospective cohort of ARC patients admitted to the hospital between June 2019 and July 2021 was enrolled as the study subjects. Concurrently, age-matched healthy individuals undergoing routine physical examinations at the hospital during the same period were recruited as the control group. Baseline demographic and clinical characteristics were compared between the two groups. Serum levels of Sirt1-6 and oxidative stress markers including malondialdehyde(MDA), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), and total antioxidant capacity(TAC)were compared. Pearson correlation analysis was performed to assess the associations between serum Sirt1-6 levels and oxidative stress parameters. Multivariate Logistic regression analysis was conducted to analysis risk factors associated with ARC occurrence. Receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of serum Sirt1-6 levels and oxidative stress indicators for ARC.RESULTS: The study included 127 ARC patients, comprising 68 men and 59 women, with a mean age of 66.05±2.58 y; the concurrent control group consisted of 121 patients, comprising 63 men and 58 women, with a mean age of 65.54±2.86 y. Serum levels of Sirt1, 3 and 6, SOD, GSH-Px, and TAC were significantly lower in the ARC group compared to the control group(all P<0.001), whereas MDA levels were markedly elevated(P<0.001). Pearson correlation analysis revealed that serum levels of Sirt1, Sirt3, Sirt6 in the ARC group were positively correlated with SOD, GSH-Px, and TAC, and negatively correlated with MDA. Multivariate Logistic regression analysis demonstrated that serum Sirt1, Sirt6, MDA, SOD, and GSH-Px were significantly associated with the occurrence of ARC(all P<0.001). ROC curve analysis showed that the combination of Sirt1, Sirt6, MDA, SOD, and GSH-Px yielded an area under the curve(AUC)of 0.995 for diagnosing ARC, which was significantly higher than that of Sirt1 alone(Z=4.978,P<0.001), Sirt6 alone(Z=7.487,P<0.001), MDA alone(Z=6.449,P<0.001), SOD alone(Z=5.773,P<0.001), or GSH-Px alone(Z=5.056,P<0.001), indicating superior diagnostic accuracy of the multimarker panel(P<0.05). CONCLUSION: Serum levels of Sirt1, 3, and 6 are generally reduced in ARC patients and are closely associated with oxidative stress imbalance. Specifically, decreased serum levels of Sirt1 and Sirt6, along with oxidative stress markers(elevated MDA and reduced SOD and GSH-Px)are identified as risk factors for ARC. Moreover, the combined detection of these indicators presents high diagnostic value for ARC.

In Weisheng Baojian (Treasures on Health), the compatibility of Qihai (CV6) and Zusanli (ST36) was widely used in the medical cases treated with acupuncture and moxibustion. Through analyzing the medical cases and summarizing the rules of acupoint compatibility, it is found that LUO Tianyi's idea of moxibustion at these two acupoints was different from the other medical masters. He combined these two acupoints in line with "spleen and stomach theory", rather than from the perspective of the direct relationship of acupoints with syndromes and zangfu organs. It reflects the diversity in the understanding of the human body and diseases among medical masters. Therefore, in order to analyze the representative understandings of ancient medical masters on the connotation and rules of acupuncture and moxibustion, we cannot simply focus on the summary from acupuncture-moxibustion masters. It is suggested that the analysis should be expended in the entire field of traditional Chinese medicine to explore the general rules of treatment with acupuncture and moxibustion, investigate the differences, and deepen the basic reasons of the distinction.
Moxibustion/history* ; Humans ; Acupuncture Points ; Acupuncture Therapy/history* ; China ; History, Ancient ; Medicine in Literature ; Medicine, Chinese Traditional/history*

With the technological innovation and the advances in information technology, acupuncture-moxibustion is on the path of modernization and high-quality development. The research on ancient acupuncture-moxibustion literature has been gradually transformed from traditional sorting and digital research to intelligent knowledge services, so as to realize the deep integration of ancient acupuncture-moxibustion knowledge with the needs of modern clinical practice and scientific research. Guided by the characteristics of acupuncture-moxibustion knowledge and based on the knowledge meta-theory, the in-depth analytical indexing and knowledge organization are conducted on more than 400 kinds of ancient acupuncture-moxibustion literature. Taking ancient literature of meridian symptoms/manifestations as an example, thematic literature research and database construction are carried out. Integrated with database, cloud platform, knowledge domain mapping and other technologies, the sharing service platform of ancient acupuncture-moxibustion knowledge is constructed. As a result, the research and development achievements can be adopted by the researchers in the field of basic theory, clinical practice and research of acupuncture-moxibustion. Finally, through the way of "digital reconstruction + intelligent application", the path and paradigm of digital research of ancient acupuncture-moxibustion literature are explored to provide the references for the innovative utilization of ancient acupuncture-moxibustion literature.
Moxibustion/history* ; Humans ; Acupuncture Therapy/history* ; History, Ancient ; Knowledge

Through the investigation of relevant literature, the concepts, methods, languages and tools of ontology were explored, and the suitable methods and tools for the ontology construction of acupoint anatomy were selected. The current mainstream anatomical ontology and related ontology of TCM were investigated so as to provide the reference for the ontology construction of acupoint anatomy. According to the knowledge attributes of acupoint anatomy, the foundational model of anatomy (FMA) was served as the reusable ontology, and in association with the attribute classification of traditional Chinese medicine language system (TCMLS), the construction approach to acupoint anatomical ontology was explored. By taking "anatomical entity of acupoints" as the top-level concept, the demonstrative study on the anatomical ontology construction was conducted on the acupoints of lung meridian of hand-taiyin.
Acupuncture Points ; Humans ; Meridians ; Medicine, Chinese Traditional ; Biological Ontologies

OBJECTIVE: To organize and display systematically the ancient medical records of stroke treated with acupuncture and moxibustion based on the knowledge element theory of information technology, so as to provide the path and paradigm for the construction of ancient acupuncture and moxibustion knowledge model. METHODS: The medical records of stroke treated with acupuncture and moxibustion were collected from the monographs of acupuncture and moxibustion and tuina, medical reports, the ancient works of traditional Chinese medicine of comprehensive collection and clinical disorders of each medical department, from the pre-Qin period to the late Qing Dynasty, collected in Zhonghua Yidian (Canon of Chinese Medicine), the fifth edition. Using "knowledge processing platform of ancient Chinese medicine books", the medical records of stroke treated with acupuncture and moxibustion in ancient time were deeply analyzed and indexed. With the MS SQL Server database adopted, the indexing results were exported into logical data; and Neo4j database was employed to build the knowledge graph of stroke treatment with acupuncture and moxibustion in ancient time. RESULTS: There were 43 medical records in 18 ancient books that met the inclusion criteria, and a logical structure was organized and composed of 65 knowledge bodies, 462 knowledge elements, 1,413 semantic types and 315 semantic associations. CONCLUSION Based on the knowledge element theory, the medical records of stroke treated with acupuncture and moxibustion in ancient time have been explored, and the logical data formed can accurately reflect the knowledge of the different attributes inside these medical records. It displays the knowledge organization category from the overall to the local. The knowledge graph generated according to the logical data is conducive to presenting the ancient acupuncture knowledge in view of the "vertical and horizontal" dimensions.
Moxibustion/history* ; Humans ; Acupuncture Therapy/history* ; Stroke/history* ; History, Ancient ; Medical Records ; China

L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.
Citrulline/biosynthesis* ; Escherichia coli/genetics* ; Metabolic Engineering/methods* ; Fermentation ; NADP/biosynthesis*

Antibiotic adjuvants offer a promising strategy for restoring antibiotic sensitivity, expanding antibacterial spectra, and reducing required dosages. Previously, compound 15 was identified as a potential adjuvant for Polymyxin B (PB) against multidrug-resistant (MDR) Pseudomonas aeruginosa DK2; however, its clinical utility was hindered by high cytotoxicity, uncertain in vivo efficacy, and an unclear synergetic mechanism. To address these challenges, we synthesized and evaluated a series of novel benzamide derivatives, with A22 emerging as a particularly promising candidate. A22 demonstrated potent synergistic activity to PB, minimal cytotoxicity, improved water solubility, and broad-spectrum synergism of polymyxins against various clinically isolated MDR Gram-negative strains. In vivo studies using Caenorhabditis elegans and mouse models further confirmed the efficacy of A22. Moreover, A22 effectively suppressed the development of PB resistance in Pseudomonas aeruginosa DK2. Mechanistic investigations revealed that A22 enhances polymyxins activity by inducing reactive oxygen species production, reducing ATP levels, increasing NOX activity, and inhibiting biofilm formation, leading to bacterial death. These findings position A22 as a highly promising candidate for the development of polymyxin adjuvants, offering a robust approach to combating MDR Gram-negative bacterial infections.

The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.