ObjectiveTo investigate the cardioprotective effects of ginsenoside Rg₂ in regulating the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway following acute myocardial infarction （AMI） in rats. Methods（1） Cellular experiment： Cardiomyocytes were isolated from 24-hour-old Sprague-Dawley （SD） neonatal rats and subjected to primary culture. An in vitro model of cardiomyocytes under an ischemic-hypoxic microenvironment was established. Cardiomyocytes were pretreated with ginsenoside Rg₂ （1， 2， 3 mg·L^-1） for 4 hours， then placed in RPMI 1640 serum-free medium and cultured for 24 hours in a three-gas incubator （94% N₂， 5% CO₂， 1% O₂）. The survival rate of cardiomyocytes was assessed using the methyl thiazolyl terazolium （MTT） assay. The levels of lactate dehydrogenase （LDH） leakage， superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） activity， and malondialdehyde （MDA） content in the cell culture supernatant were measured using spectrophotometry. （2） Animal experiment： Specific-pathogen-free （SPF） SD rats were used to establish an AMI model using the Olivette method combined with previous studies. Rats that survived 24 hours post-surgery were randomly divided into a model group and ginsenoside Rg₂ high-， medium-， and low-dose groups. The normal and model groups received normal saline， while the ginsenoside Rg₂ groups were administered intragastrically at doses of 8， 4， and 2 mg·kg^-1， once daily for 3 days. The levels of SOD， MDA， and GSH-Px in myocardial tissues were detected. Cardiomyocyte apoptosis was assessed using the TdT-mediated dUTP biotin nick end labeling （TUNEL） assay. The mRNA and protein expression levels of PI3K， Akt， mTOR， p62， nuclear factor-κB p65 （NF-κB p65）， and microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/Ⅰ in myocardial tissues were analyzed using real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. Pathological changes in the infarct border zone were observed under a light microscope. Results（1） Cellular experiment： Compared with the normal group， the model group exhibited a significantly decreased cardiomyocyte survival rate， as well as reduced SOD and GSH-Px activity， whereas LDH activity and MDA content were significantly increased （P<0.05）. Compared with the model group， ginsenoside Rg₂ intervention significantly increased cardiomyocyte survival， SOD activity， and GSH-Px activity， while reducing LDH activity and MDA content （P<0.05） in a dose-dependent manner. Pathological examination revealed that ginsenoside Rg₂ alleviated infarct size， myocardial degeneration， and necrosis， while significantly reducing cardiomyocyte apoptosis. （2） Animal experiment： Compared with the normal group， the model group exhibited significantly lower SOD and GSH-Px activity （P<0.05） and higher MDA content （P<0.05） in myocardial tissues. Compared with the model group， all ginsenoside Rg₂ groups showed significantly increased SOD and GSH-Px activity （P<0.05） and reduced MDA content （P<0.05）. Compared with the normal group， the model group exhibited significantly decreased mRNA and protein expression levels of PI3K， Akt， mTOR， p62，and LC3 Ⅱ/Ⅰ，whereas the expression levels of NF-κB p65 were significantly increased （P<0.05）. Compared with the model group， the ginsenoside Rg₂ groups showed significantly increased PI3K， Akt， mTOR， and p62 expression， while NF-κB p65 expression levels were significantly decreased （P<0.05） in a dose-dependent manner，the mRNA and protein expression levels of LC3 Ⅱ/Ⅰ in ginsenoside Rg₂ high-dose groups were significantly increased（P<0.05）. ConclusionGinsenoside Rg₂ exerts cardioprotective effects following AMI in rats， potentially through the regulation of PI3K/Akt/mTOR-related protein expression.

Diffuse large B-cell lymphoma （DLBCL） is a highly heterogeneous disease. Although standard first-line regimens can cure ＞50% of patients， approximately one-third of them develop relapsed/refractory DLBCL （r/r DLBCL）. Consequently， immunotherapy targeting molecular abnormalities has become pivotal for managing r/r DLBCL. The results of this review show that with advances in understanding DLBCL pathogenesis and the tumor immune microenvironment， antibody-based therapies have evolved rapidly， progressing from monoclonal antibodies （e.g.， rituximab， tafasitamab） to bispecific antibodies（e.g.， odronextamab，glofitamab， epcoritamab） and antibody-drug conjugate （e.g.， polatuzumab vedotin， loncastuximab tesirine）. These engineered agents enhance immune cytotoxicity and tumor-specific targeting， providing novel therapeutic options for r/r DLBCL patients.

OBJECTIVE: To observe the effect of electroacupuncture combined with enhanced recovery after surgery (ERAS) protocol on promoting intestinal function in patients after gastric cancer surgery. METHODS: Forty-four patients who underwent radical gastrectomy for gastric cancer were randomly divided into an experimental group (22 cases, 3 cases were excluded) and a control group (22 cases, 4 cases were excluded). Both groups received treatment under ERAS protocol, the experimental group was given electroacupuncture at bilateral Neiguan (PC6), Hegu (LI4), Zusanli (ST36) and Quchi (LI11), disperse-dense wave was selected, with frequency of 2 Hz/100 Hz. The control group received placebo electroacupuncture intervention, with the same acupoints as the experimental group, electrode pads were placed on the acupoints without electrical stimulation. Each session lasted 30 min, starting from 1 h after surgery, once every 24 h, until the patient resumed anal flatus. The intestinal sound rate of both groups was observed 24 h before surgery and 24, 48 h after surgery. The bowel sound recovery time (BSRT), time to first anal flatus, time to first defecation, and tolerance to oral enteral nutrition suspension were compared between the two groups. The levels of serum C-reactive protein (CRP), interleukin (IL)-2, IL-4, IL-6, IL-10, IL-12, IL-17, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were measured 24 h before surgery and 24 h after surgery in both groups. RESULTS: The intestinal sound rate 24 h after surgery was decreased compared with that 24 h before surgery in the two groups (P<0.05), the intestinal sound rate 24, 48 h after surgery in the experimental group was higher than that in the control group (P<0.05). The BSRT in the experimental group was earlier than that in the control group (P<0.05) .The levels of serum CRP, IL-6, IL-10 24 h after surgery in the experimental group were higher than those 24 h before surgery (P<0.05), while the levels of serum CRP, IL-4, IL-6, IL-10, IFN-γ in the control group were higher than those 24 h before surgery (P<0.05); the levels of serum CRP、IL-4、IFN-γ 24 h after surgery in the experimental group were lower than those in the control group (P<0.05) .The tolerance rate of oral enteral nutrition suspension in the experimental group was 84.2% (16/19), which was higher than 50.0% (9/18) in the control group (P<0.05). CONCLUSION Electroacupuncture combined with ERAS protocol can improve the intestinal motility, shorten the BSRT, enhance the tolerance of oral intake, and reduce inflammatory response in patients after gastric cancer surgery.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Acupuncture Points ; C-Reactive Protein/metabolism* ; Electroacupuncture ; Gastrectomy ; Interleukin-10 ; Interleukin-6 ; Intestines/physiopathology* ; Stomach Neoplasms/therapy*

Objective To explore the prognostic value of glycolysis-related genes in colorectal cancer (CRC) patients and formulate a novel glycolysis-related prognostic risk model. Methods Single-cell and bulk transcriptomic data of CRC patients, along with clinical information, were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Glycolysis scores for each sample were calculated using single-sample Gene Set Enrichment Analysis (ssGSEA). Kaplan-Meier survival curves were generated to analyze the relationship between glycolysis scores and overall survival. Novel glycolysis-related subgroups were defined among the cell type with the highest glycolysis scores. Gene enrichment analysis, metabolic activity assessment, and univariate Cox regression were performed to explore the biological functions and prognostic impact of these subgroups. A prognostic risk model was built and validated based on genes significantly affecting the prognosis. Gene Set Enrichment Analysis (GSEA) was conducted to explore differences in biological processes between high- and low-risk groups. Differences in immune microenvironment and drug sensitivity between these groups were assessed using R packages. Potential targeted agents for prognostic risk genes were predicted using the Enrichr database. Results Tumor tissues showed significantly higher glycolysis scores than normal tissues, which was associated with a poor prognosis in CRC patients. The highest glycolysis score was observed in epithelial cells, within which we defined eight novel glycolysis-related cell subpopulations. Specifically, the P4HA1⁺ epithelial cell subpopulation was associated with a poor prognosis. Based on signature genes of this subpopulation, a six-gene prognostic risk model was formulated. GSEA revealed significant biological differences between high- and low-risk groups. Immune microenvironment analysis demonstrated that the high-risk group had increased infiltration of macrophages and tumor-associated fibroblasts, along with evident immune exclusion and suppression, while the low-risk group exhibited higher levels of B cell and T cell infiltration. Drug sensitivity analysis indicated that high-risk patients were more sensitive to Abiraterone, while low-risk patients responded to Cisplatin. Additionally, Valproic acid was predicted as a potential targeted agent. Conclusion High glycolytic activity is associated with a poor prognosis in CRC patients. The novel glycolysis-related prognostic risk model formulated in this study offers significant potential for enhancing the diagnosis and treatment of CRC.
Humans ; Colorectal Neoplasms/pathology* ; Glycolysis/genetics* ; Prognosis ; Transcriptome ; Tumor Microenvironment/genetics* ; Gene Expression Profiling ; Single-Cell Analysis ; Gene Expression Regulation, Neoplastic ; Male ; Female ; Kaplan-Meier Estimate

Metastatic dissemination is the major cause of death from breast-cancer (BC). Fusobacterium nucleatum (F.n) is widely enriched in BC and has recently been identified as one of the high-risk factors for promoting BC metastasis. Here, with an experimental model, we demonstrated that intratumoral F.n induced BC aggressiveness by transcriptionally activating Epithelial-mesenchymal transition-associated genes. Therefore, the F.n may be a potential target to prevent metastasis. Given the fact that cancer-associated fibroblasts (CAFs) are abundant in BC and located near blood vessels, we report an optogenetic system that drives CAF to in situ produce human antibacterial peptide LL37, with the characteristics of biosafety and freely intercellular trafficking, for depleting intratumoral F.n, leading to a 72.1% reduction in lung metastatic nodules number without affecting the balance of the systemic flora. Notably, mild photothermal treatment was found that could normalize CAF, contributing to synergistically inhibiting BC metastasis. In addition, the system can also simultaneously encode a gene of TNF-related apoptosis-inducing ligand to suppress the primary tumor. Together, our study highlights the potential of local elimination of tumor pathogenic bacteria to prevent BC metastasis.

Biosensors have become powerful tools for real-time monitoring of specific small molecules and precise control of gene expression in biological systems. High-throughput sensors for 1, 4-butanediamine biosynthesis can greatly improve the screening efficiency of high-yielding 1, 4-butanediamine strains. However, the strategies for adapting the characteristics of biosensors are still rarely studied, which limits the applicability of 1, 4-butanediamine biosensors. In this paper, we propose the development of a 1, 4-butanediamine biosensor based on the transcriptional regulator PuuR, whose homologous operator puuO is installed in the constitutive promoter P_gapA of Escherichia coli to control the expression of the downstream superfolder green fluorescent protein (sfGFP) as the reporter protein. Finally, the biosensor showed a stable linear relationship between the GFP/OD₆₀₀ value and the concentration of 1, 4-butanediamine when the concentration of 1, 4-butanediamine was 0-50 mmol/L. The promoters with different strengths in the E. coli genome were used to modify the 1, 4-butanediamine biosensor, and the functional properties of the PuuR-based 1, 4-butanediamine biosensor were explored and improved, which laid the groundwork for high-throughput screening of engineered strains highly producing 1, 4-butanediamine.
Biosensing Techniques/methods* ; Escherichia coli/metabolism* ; Promoter Regions, Genetic/genetics* ; Green Fluorescent Proteins/metabolism* ; Transcription Factors/genetics* ; Escherichia coli Proteins/genetics* ; Diamines/metabolism* ; Gene Expression Regulation, Bacterial

Objective To determine the reference red blood cells with weak agglutination intensity of low positive quali-ty control products by comparing RhD antigen expression intensity difference according to the serological results.Methods The RhD(+)red blood cells were detected by microcolumn gel method with 1 500 times diluted anti-D typing reagent.The samples with weak and strong RhD antigen expression intensity were selected as the reference red blood cells for weak agglu-tination intensity of low positive quality control products,and verification was performed.Results Ten RhD(+)red blood cells were detected with diluted anti-D typing reagent,of which 8 were 1+and 2 were±.Red blood cells with agglutination intensity of 1+were used as the benchmark to determine the maximum dilution ratio of anti-D typing reagent when their ag-glutination intensity was 1+.As the preparation standard of low positive quality control products,the agglutination intensity of red blood cells with low RhD antigen expression intensity was extremely weak±,which was difficult to ensure the stability of its control limit properties.Based on red blood cells with agglutination intensity of±,the maximum dilution ratio of anti-D typing reagent with agglutination intensity of 1+was re-determined as the preparation standard of low positive quality con-trol products,and the results met the requirements of quality control product setting.Conclusion Using red blood cells with low RhD antigen expression intensity as the benchmark to set the weak agglutination intensity of the low positive quality control products can avoid the loss of control due to the low target value.

Background::Pathological scars are a disorder that can lead to various cosmetic, psychological, and functional problems, and no effective assessment methods are currently available. Assessment and treatment of pathological scars are based on cutaneous manifestations. A two-photon microscope (TPM) with the potential for real-time non-invasive assessment may help determine the under-surface pathophysiological conditions in vivo. This study used a portable handheld TPM to image epidermal cells and dermal collagen structures in pathological scars and normal skin in vivo to evaluate the effectiveness of treatment in scar patients. Methods::Fifteen patients with pathological scars and three healthy controls were recruited. Imaging was performed using a portable handheld TPM. Five indexes were extracted from two dimensional (2D) and three dimensional (3D) perspectives, including collagen depth, dermo-epidermal junction (DEJ) contour ratio, thickness, orientation, and occupation (proportion of collagen fibers in the field of view) of collagen. Two depth-dependent indexes were computed through the 3D second harmonic generation image and three morphology-related indexes from the 2D images. We assessed index differences between scar and normal skin and changes before and after treatment.Results::Pathological scars and normal skin differed markedly regarding the epidermal morphological structure and the spectral characteristics of collagen fibers. Five indexes were employed to distinguish between normal skin and scar tissue. Statistically significant differences were found in average depth ( t = 9.917, P <0.001), thickness ( t = 4.037, P <0.001), occupation ( t= 2.169, P <0.050), orientation of collagen ( t = 3.669, P <0.001), and the DEJ contour ratio ( t = 5.105, P <0.001). Conclusions::Use of portable handheld TPM can distinguish collagen from skin tissues; thus, it is more suitable for scar imaging than reflectance confocal microscopy. Thus, a TPM may be an auxiliary tool for scar treatment selection and assessing treatment efficacy.

Objective Developing a new type of full-face respiratory protective mask for nuclear facility sites to enhance the sound transmission function and improve the facial adaptability. Methods Combined with feedback from on-site practical needs, this study utilized finite element simulation and ergonomic design methods to investigate the voice transmission units of full-face masks and the facial features of workers at key nuclear facilities. Based on the research results, a new full-face respiratory protection mask structure was designed. Results The optimized structure of passive thin film voice transmission unit significantly enhanced voice transmission efficiency, reducing average voice transmission loss by approximately 70% compared to the control group using thin plate units of equivalent thickness. The existing facial feature test panels insufficiently cover and unevenly classify the facial features of workers at key nuclear facilities. In this study, a specialized test panel based on measurement data achieved a total coverage of 98.5% with high distribution uniformity within each class, providing effective guidance for redesigning full-face mask structural parameters. In comparison to foreign products currently utilized in nuclear facilities, the newly designed full-face mask structure exhibited excellent tightness and structural safety and reliability, and can be cleaned, decontaminated, and reused. Conclusion The results of this study provide significant guidance for improving and optimizing full-face respiratory protection mask used at nuclear facilities, as well as promoting domestic production of high-quality full-face respiratory protection masks.

Objective:To clarify the clinicopathological features, prognosis, and recurrence pattern of early-onset gastric cancer (EOGC).Methods:Using data from the gastric cancer database of Zhongshan Hospital, Fudan University, we performed a retrospective, large-scale, real-world study of 5046 patients with gastric cancer who had undergone redical or palliative gastrectomy from January 2013 to December 2018, including 425 patients with EOGC (age ≤45 years) and 4621 controls. All those patients were pathologically confirmed adenocarcinoma with complete follow-up of five years. Residue gastric cancer and patients without complete clinical or follow-up data were excluded. We used a combination of outpatient and telephone follow-up, ending in October 2022 (median duration of follow-up 60 months), and compared the clinicopathological features and prognosis of the two groups.Results:The clinicopathological features of EOGC included female predominance (61.1% [262/425 vs. 26.3% [1217/4621], χ 2=234.215, P<0.001), fewer comorbidities (31.3% [133/425] vs. 58.5% [2703/4621], χ 2=34.378, P<0.001), poorer differentiation (90.6% [385/425] vs. 78.2% [3614/4621], χ 2=30.642, P<0.001), higher proportion of diffuse type (53.9% [229/425] vs. 18.3% [846/4621], χ 2=274.474, P<0.001), higher proportion of T4 stage (44.7% [190/425] vs. 37.5% [1733/4621], χ 2=17.535, P=0.001), more lymph node metastases (60.5% [257/425] vs. 53.9% [2491/4621], χ 2=6.764, P=0.009), and higher proportion of pathological stage III/IV (47.5% [202/425] vs. 42.4% [1959/4621], χ 2=4.093, P=0.043). The 5-year overall survival rates of the EOGC and control groups were 55.1% and 49.1%, respectively. Overall survival was significantly better in the EOGC than in the control group ( P<0.001). According to subgroup analysis, the prognosis of pathological stage I/II/III EOGC was better than that of the control group. Recurrence rates were similar in the two groups, whereas patients with EOGC had a higher proportion of peritoneal recurrence (7.8% [33/425] vs. 3.2% [146/4621], χ 2=23.741, P<0.001) and a lower proportion of distant metastasis (4.9% [21/425] vs. 8.3% [385/4621], χ 2=6.247, P=0.012). Conclusion:EOGC has unique clinicopathological features and recurrence patterns and resectable EOGC has a better prognosis, suggesting that patients with EOGC should be actively treated with the focus on preventing peritoneal recurrence.