BACKGROUND: Retinal ganglion cell (RGC) death caused by acute ocular hypertension is an important characteristic of acute glaucoma. Receptor-interacting protein kinase 3 (RIPK3) that mediates necroptosis is a potential therapeutic target for RGC death. However, the current understanding of the targeting agents and mechanisms of RIPK3 in the treatment of glaucoma remains limited. Notably, artificial intelligence (AI) technologies have significantly advanced drug discovery. This study aimed to discover RIPK3 inhibitor with AI assistance. METHODS: An acute ocular hypertension model was used to simulate pathological ocular hypertension in vivo . We employed a series of AI methods, including large language and graph neural network models, to identify the target compounds of RIPK3. Subsequently, these target candidates were validated using molecular simulations (molecular docking, absorption, distribution, metabolism, excretion, and toxicity [ADMET] prediction, and molecular dynamics simulations) and biological experiments (Western blotting and fluorescence staining) in vitro and in vivo . RESULTS: AI-driven drug screening techniques have the potential to greatly accelerate drug development. A compound called HG9-91-01, identified using AI methods, exerted neuroprotective effects in acute glaucoma. Our research indicates that all five candidates recommended by AI were able to protect the morphological integrity of RGC cells when exposed to hypoxia and glucose deficiency, and HG9-91-01 showed a higher cell survival rate compared to the other candidates. Furthermore, HG9-91-01 was found to protect the retinal structure and reduce the loss of retinal layers in an acute glaucoma model. It was also observed that the neuroprotective effects of HG9-91-01 were highly correlated with the inhibition of PANoptosis (apoptosis, pyroptosis, and necroptosis). Finally, we found that HG9-91-01 can regulate key proteins related to PANoptosis, indicating that this compound exerts neuroprotective effects in the retina by inhibiting the expression of proteins related to apoptosis, pyroptosis, and necroptosis. CONCLUSION AI-enabled drug discovery revealed that HG9-91-01 could serve as a potential treatment for acute glaucoma.
Animals ; Glaucoma/metabolism* ; Neuroprotective Agents/pharmacology* ; Mice ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism* ; Artificial Intelligence ; Retinal Ganglion Cells/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation ; Mice, Inbred C57BL ; Male

Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

Pyroptosis is an identified programmed cell death that has been highly linked to endoplasmic reticulum (ER) dynamics. However, the crucial proteins for modulating dynamic ER membrane curvature change that trigger pyroptosis are currently not well understood. In this study, a biotin-labeled chemical probe of potent pyroptosis inducer α-mangostin (α-MG) was synthesized. Through protein microarray analysis, reticulon-4 (RTN4/Nogo), a crucial regulator of ER membrane curvature, was identified as a target of α-MG. We observed that chemically induced proteasome degradation of RTN4 by α-MG through recruiting E3 ligase UBR5 significantly enhances the pyroptosis phenotype in cancer cells. Interestingly, the downregulation of RTN4 expression significantly facilitated a dynamic remodeling of ER membrane curvature through a transition from tubules to sheets, consequently leading to rapid fusion of the ER with the cell plasma membrane. In particular, the ER-to-plasma membrane fusion process is supported by the observed translocation of several crucial ER markers to the "bubble" structures of pyroptotic cells. Furthermore, α-MG-induced RTN4 knockdown leads to pyruvate kinase M2 (PKM2)-dependent conventional caspase-3/gasdermin E (GSDME) cleavages for pyroptosis progression. In vivo, we observed that chemical or genetic RTN4 knockdown significantly inhibited cancer cells growth, which further exhibited an antitumor immune response with anti-programmed death-1 (anti-PD-1). In translational research, RTN4 high expression was closely correlated with the tumor metastasis and death of patients. Taken together, RTN4 plays a fundamental role in inducing pyroptosis through the modulation of ER membrane curvature remodeling, thus representing a prospective druggable target for anticancer immunotherapy.
Pyroptosis/immunology* ; Humans ; Endoplasmic Reticulum/immunology* ; Animals ; Nogo Proteins/antagonists & inhibitors* ; Mice ; Cell Line, Tumor ; Xanthones/pharmacology* ; Neoplasms/pathology* ; Mice, Nude

Objective:To analyze the trends in refractive error and ocular biological parameters in elementary school students over 5 years, and to investigate the patterns of change before and after myopia onset.Methods:A cohort study was adopted.A total of 1 986 first-grade students from the Anyang Childhood Eye Study were enrolled in this cohort study and their right eye data were taken for analysis, including 1 126 boys and 860 girls.Every year, cycloplegic autorefraction was performed with 1% cyclopentolate eyedrops to obtain the spherical equivalent (SE).The axial length (AL), anterior chamber depth, lens thickness, mean corneal curvature (Km) and other parameters were obtained by ocular biometry.The lens refractive power (LP) was calculated using the Bennett formula.The subjects were assigned to persistent myopia group, non-myopia group and new onset myopia group.According to the age of myopia onset, the new onset myopia group was subdivided into the 8-, 9-, 10-, 11- and 12-year-old myopia groups to compare the differences in refractive error and ocular bioparameters among groups at different time points of follow-up.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.TRECKY2018-030).Written informed consent form was obtained from the guardians of each subject.Results:All children had a gradual SE drift toward myopia and a gradual increase in the AL with age, and there were significant differences in SE and AL between adjacent follow-up ages within the three groups (all at P<0.05).The earlier the onset of myopia, the higher the myopia SE and the longer the AL of the eye at the same follow-up age, the differences in SE between adjacent groups were statistically significant (all at P<0.05), and the differences in AL between adjacent groups at the follow-up age of 8 to 12 years were statistically significant (all at P<0.05).In the nonmyopia group, SE drifted toward emmetropia at a slow and steady rate of (-0.23±0.27)D/year, and AL also increased slowly and steadily at (0.18±0.13)mm/year.In the new onset myopia group, the changes in SE in the third, second, and first years before myopia onset were (-0.32±0.25), (-0.45±0.33), and (-0.98±0.44)D, and the increases in AL were (0.25±0.12), (0.32±0.15), and (0.48±0.19)mm, respectively.Both SE and AL change rates began to accelerate before myopia onset and slowed down after myopia onset, with statistically significant differences in the overall comparison of SE and AL change rates at different time intervals before and after myopia onset (all at P<0.001).The AL at myopia onset in boys was (24.11±0.70)mm, which was longer than (23.60±0.66)mm in girls ( t=159.71, P<0.01).LP decreased with age in all groups, with a faster rate before the age of 9 years and a slower rate after the age of 9 years.The mean decrease rate in LP was (-0.48±0.19), (-0.44±0.20), (-0.49±0.16), (-0.51±0.18), and (-0.48±0.19)D/year in the persistent myopia group and 8~11-year-old myopia group, respectively, which were significantly faster than -0.42±0.17 D/year in 12-year-old myopia group and (0.37±0.15)D/year in nonmyopia group (all at P<0.05).There was no statistically significant difference in Km among groups at different follow-up ages (all at P>0.05). Conclusions:The AL begins to grow at an accelerated rate 3 years before myopia onset, and the increase rate of the AL slows down after the onset of myopia, but it is still significantly faster than that of non-myopic children.In this process, the decrease in LP plays a compensatory role; there is no significant change in corneal curvature.The AL of males at the onset of myopia is longer than that of females at the same age.AL is an important indicator for the prevention and control of myopia.It is important to consider gender differences and to pay more attention to the growth rate when assessing AL.

Objective To explore and evaluate the micro-elimination mode of hepatitis C virus(HCV)in patients in a general hospital in Jiangxi Province,and provide reference for formulating provincial strategies to eliminate the public health hazards of HCV.Methods Relevant data of hospitalized patients who received HCV screening at the hospital from January 2021 to December 2022 were collected.Data from 2021 were set as the baseline and data from 2022 as the HCV micro-elimination mode operating data.Indexes such as gender,age,department,anti-HCV posi-tive rate,HCV-RNA positive rate,etc.were analyzed.The operational effect was analyzed based on specialty de-partment visiting rate,treatment rate,and loss-to-follow-up(LTFU)rate.The development of the mode was quan-titatively evaluated by superiority weakness opportunity threats-analytic hierarchy process(SWOT-AHP)method.Results A total of 397 744 hospitalized patients underwent anti-HCV screening from 2021 to 2022,with a male to female ratio of 1.34∶1 and an average age of 54 years old.The main departments for patients receiving screening were infectious diseases/gastroenterology departments.HCV gene subtypes in specimens from 62 patients was mainly subtype 1b(43 specimens),followed by subtype 6(9 specimens).The first HCV early warning rate after mode operation was 81.20％,the anti-HCV detection rate after warning was 93.15％,and the anti-HCV detection rate after subsequent pop-up window early warning was 100％.Compared with 2021,the treatment rate of HCV hospi-talized patients in 2022 has increased,with statistically significant difference(P＜0.05).However,there were no statistically significant differences in patients'specialty department visiting rate and LTFU rate(both P＞0.05).Barycentric coordinates calculation P(X,Y)=(0.018 2,0.006 9)located in the first quadrant.Conclusion The HCV micro-elimination mode for patients in a general hospital in Jiangxi Province is feasible and effective.Further mode optimization should focus on strengths/opportunities(SO)strategy,to rely on internal advantages and utilize external opportunities.

Objective:To investigate the current status of sarcopenia and self-management in elderly patients with type 2 diabetes mellitus (T2DM), and to explore the identification of risk factors for sarcopenia by diabetes self-management ability.Methods:Using convenience sampling, 284 elderly patients with T2DM who visited a community health service center in Beijing from March to September 2023 were selected as subjects. The patients were screened for sarcopenia and received related health examinations based on Asian working group for sarcopenia (AWGS) 2019 Consensus. Surveys were conducted using general information questionnaire, the Summary of Diabetes Self-Care Activities measure, and other questionnaires. Patients were divided into groups according to the presence or absence of muscle attenuation (defined as suspected and confirmed sarcopenia).Results:The prevalence of muscle attenuation in the 284 elderly patients with T2DM was 48.9%, and the prevalence of sarcopenia was 15.9%. The proportions of females, patients who are over 70 years old, and patients with a sedentary lifestyle were significantly higher in the group with muscle attenuation compared with the group without muscle attenuation. High-quality protein intake, extremity skeletal muscle mass index, grip strength, and six-meter walking speed were significantly lower in the muscle attenuation group. Binary logistic regression analysis revealed that age, alcohol consumption, a sedentary lifestyle, and high-quality protein intake were influencing factors for sarcopenia in elderly patients with T2DM ( P<0.05). The total self-care scores and subtotals in exercise domains showed significant differences ( P<0.05) between patients with and without muscle attenuation. Univariate analysis indicated significant differences in self-management behaviors among patient groups stratified by grip strengths and 6-meter walk speeds ( P<0.05). Multivariate linear regression analysis showed that grip strength and 6-meter walk speed were influencing factors for exercise management behaviors in elderly patients with T2DM ( P<0.05). Conclusions The prevalence of sarcopenia in elderly patients with T2DM is relatively high, and their level of diabetes self-management is medium to low. Practitioners should pay extra attention to patients who are over 70 years old, with sedentary habits, with low intake of high-quality protein, and females. It's recommended to use grip strength and 6-meter walk speed tests as initial screening tools for sarcopenia in elderly patients with T2DM, in order to identify risks early and implement targeted health management to promote the development of good self-management behaviors among patients.

Purpose: The development of optics-based wearables for bladder volume monitoring has emerged as a significant topic in recent years. Given the innovative nature of this technology, there is currently no bladder phantom available to effectively validate these devices against more established gold standards, such as ultrasound. In this study, we showcase and demonstrate the performance of our hybrid bladder phantom by using an optical device and making comparisons with ultrasound. Methods: A series of validation tests, including phantom repeatability, ultrasound scanning, and an optical test, were performed. A near-infrared optical device was utilized to conduct diffuse optical spectroscopy (DOS). Machine learning models were employed to construct predictive models of volume using optical signals. Results: The size and position of an embedded balloon, serving as an analog for the bladder, were shown to be consistent when infused with 100 mL to 350 mL of water during repeatability testing. For DOS data, we present 7 types of machine learningbased models based on different optical signals. The 2 best-performing models demonstrated an average absolute volume error ranging from 12.7 mL to 19.0 mL. Conclusions In this study, we introduced a hybrid bladder phantom designed for the validation of near-infrared spectroscopy-based bladder monitoring devices in comparison with ultrasound techniques. By offering a reproducible and robust validation tool, we aim to support the advancement of next-generation optical wearables for bladder volume monitoring.

In recent years, reports of adverse reactions related to traditional Chinese medicine(TCM) have been on the rise, especially some traditionally considered "non-toxic" TCM(such as Dictamni Cortex). This has aroused the concern of scholars. This study aims to explore the metabolomic mechanism underlying the difference in liver injury induced by dictamnine between males and females through the experiment on 4-week-old mice. The results showed that the serum biochemical indexes of liver function and organ coefficients were significantly increased by dictamnine(P<0.05), and hepatic alveolar steatosis was mainly observed in female mice. However, no histopathological changes were observed in the male mice. Furthermore, a total of 48 differential metabolites(such as tryptophan, corticosterone, and indole) related to the difference in liver injury between males and females were screened out by untargeted metabolomics and multivariate statistical analysis. According to the receiver operating characteristic(ROC) curve, 14 metabolites were highly correlated with the difference. Finally, pathway enrichment analysis indicated that disorders of metabolic pathways, such as tryptophan metabolism, steroid hormone biosynthesis, and ferroptosis(linoleic acid metabolism and arachidonic acid metabolism), may be the potential mechanism of the difference. Liver injury induced by dictamnine is significantly different between males and females, which may be caused by the disorders of tryptophan metabolism, steroid hormone biosynthesis, and ferroptosis pathways.
Female ; Male ; Animals ; Mice ; Tryptophan ; Metabolomics ; Fatty Liver ; Steroids ; Hormones

OBJECTIVE: To investigate the effect of celastrol on the proliferation and apoptosis of human multiple myeloma (MM) cell lines, reveal the relationship between IRAK4/ERK/p38 signaling pathway and celastrol regulating the proliferation and apoptosis of H929 and ARP-1 cells, and explore whether celastrol combined with bortezomib has synergistic effect. METHODS: CCK-8 method was used to detect the viability of MM cell lines H929 and ARP-1 treated by different concentrations of celastrol, bortezomib, and their combination, and the synergistic effect was determined by Kim's formula. The apoptosis rate of H929 cells and necrosis rate of ARP-1 were detected by Annexin V/PI method. The expression of key proteins and apoptosis proteins in IRAK4/ERK/p38 signaling pathway were detected by Western blot. RESULTS: Celastrol could significantly inhibit the proliferation of H929 and ARP-1 cells (r=0.9018, r=0.9244) and induce apoptosis in a time-dependent manner. Compared with the control group, celastrol could significantly up-regulate the expression of PARP and cleaved caspase-3 while down-regulate the expression of p-IRAK4, p-ERK, and p-p38 in H929 and ARP-1 cells. Celastrol and bortezomib alone inhibited the proliferation of H929 and ARP-1 cells. Compared with celastrol and bortezomib alone, their combination had lower cell survival rate and higher apoptosis rate (P<0.05). CONCLUSION Celastrol can inhibit the proliferation and promote the apoptosis of H929 and ARP-1 cells, which may be related to inhibiting the phosphorylation of IRAK4 and blocking the activation of IRAK4/ERK/p38 signaling pathway. Celastrol combined with bortezomib has synergistic effect, which can more effectively inhibit the proliferation and induce apoptosis of H929 and ARP-1 cells.
Apoptosis ; Bortezomib/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Interleukin-1 Receptor-Associated Kinases ; Multiple Myeloma ; Pentacyclic Triterpenes ; Signal Transduction

OBJECTIVE: To investigate the clinical features, distribution of pathogenic bacteria, and drug resistance of bloodstream infection in children with acute leukemia. METHODS: Clinical data of 93 blood culture-positive children with acute leukemia from January 2015 to December 2019 in Department of Pediatrics, The Second Hospital of Anhui Medical University were analyzed retrospectively. RESULTS: In these 93 cases, 78 cases were in the period of neutrophil deficiency. There were 54 Gram-negative bacteria (G^-) (58.1%) found through blood culture, and the top 4 strains were Escherichia coli (15.1%), Klebsiella pneumoniae (13.9%), Pseudomonas aeruginosa (6.5%), and Enterobacter cloacae (6.5%). There were 39 Gram-positive bacteria (G⁺) (41.9%) detected, and the top 4 strains were Staphylococcus epidermidis (10.8%), Streptococcus pneumoniae (6.5%), Staphylococcus hemolyticus (5.4%), and Staphylococcus human (5.4%). Among 74 strains of pathogenic bacteria from acute lymphoblastic leukemia (ALL) children, there were 29 strains of G⁺ bacteria (39.2%) and 45 strains of G^- bacteria (60.8%). While in 19 strains from acute myeloblastic leukemia (AML) patients, G^- bacteria accounted for 47.4% and G⁺ bacteria accounted for 52.6%. In 15 ALL children without neutropenia, G⁺ bacteria made up the majority of the strains (66.7%). In the 93 strains of pathogenic bacteria, 13 (13.9%) strains were multidrug-resistant. Among them, extended-spectrum β-lactamases accounted for 42.9%, carbapenemase-resistant enzyme Klebsiella pneumoniae 15.4%, and carbapenemase-resistant enzyme Enterobacter cloacae strains 33.3%, which were detected from G^- bacteria. While, 13.3% of methicillin-resistant coagulase-negative Staphylococci accounted for 13.3% detected from G⁺ bacteria, but linezolid, vancomycin, teicoplanin Staphylococcus and Enterococcus resistant were not found. The average procalcitonin (PCT) value of G^- bacteria infection was (11.02±20.282) ng/ml, while in G⁺ infection it was (1.81±4.911) ng/ml, the difference was statistically significant (P<0.05). The mean value of C-reactive protein (CRP) in G^- infection was (76.33±69.946) mg/L, and that in G⁺ infection was (38.34±57.951) mg/L. The prognosis of active treatment was good, and only one case died of septic shock complicated with disseminated intravascular coagulation (DIC) and gastrointestinal bleeding caused by carbapenemase-resistant enzyme enterobacteriaceae. CONCLUSION G^- is the major bacteria in acute leukemia children with bloodstream infection, but the distribution of ALL and AML strains is different. G^- bacteria dominates in ALL, while G⁺ bacteria and G^- bacteria are equally distributed in AML. Non-agranulocytosis accompanied by bloodstream infections is dominant by G⁺ bacteria. The mean value of PCT and CRP are significantly higher in G^- bacteria infection than in G⁺ bacteria.
Acute Disease ; Anti-Bacterial Agents/therapeutic use* ; Bacteremia/microbiology* ; Bacteria ; Child ; Drug Resistance, Bacterial ; Humans ; Leukemia, Myeloid, Acute/drug therapy* ; Microbial Sensitivity Tests ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Procalcitonin ; Retrospective Studies ; Sepsis/drug therapy*