Objective: To investigate the role of butyric acid-producing bacteria in long bone homeostasis in mice with periodontitis under a high-fat/high-sugar diet and to provide new insights for the prevention and treatment of periodontitis and related bone metabolic diseases. Methods: This study has been approved by the Animal Welfare and Ethics Committee of the Experimental Animal Center. Initially, 14 mice were randomly divided into the CON group (the control group) and the LIG group (the periodontitis group). Mice in the LIG group had experimental periodontitis induced by ligating the second maxillary molars bilaterally and were fed a high-fat and high-sugar diet. After 8 weeks, samples were collected. Micro-computed tomography (Micro-CT) was used to analyze alveolar bone resorption and various parameters of the proximal tibia trabecular bone, including bone mineral density (BMD), bone volume per tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). After decalcification, hematoxylin and eosin (HE) staining was performed on maxillary bone sections to assess periodontal tissue inflammation and connective tissue destruction. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect related genes in the distal femur and proximal tibia bone tissues, including osteocalcin (OCN), osteogenic transcription factor (Osterix), osteoprotegerin (OPG), tartrate resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), receptor activator of nuclear factor kappa-B (RANK), and receptor activator of nuclear factor kappa-B ligand (RANK-L). Subsequently, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + butyric acid-producing bacteria (BP) group, and LIG + BP group. The breeding, sampling, and sample detection methods remained the same. Finally, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + sodium butyrate (SB) group, and LIG + SB group. The breeding, sampling, and sample detection methods remained the same. Results: ①Periodontitis modeling was successful. Compared with the CON group, the LIG group exhibited significant alveolar bone resorption of the maxillary second molar, aggravated periodontal tissue inflammation, and connective tissue destruction. ②Periodontitis exacerbated long bone resorption in mice fed a high-fat high-sugar diet. Compared with the CON group, the LIG group had significantly lower BMD, BV/TV, Tb.N, and Tb.Th (P<0.05), and significantly higher Tb.Sp (P<0.05). HE staining of the proximal tibia showed that the trabeculae in the LIG group were sparse and disordered, with some areas showing fractures or dissolution. The expression of osteoblast markers (OCN, Osterix, OPG) was significantly lower in the LIG group (P<0.05), while the expression of the osteoclast marker TRAP showed an increasing trend (P>0.05). The ratio of RANK-L/OPG was significantly higher in the LIG group compared with the CON group (P<0.05). ③ Supplementation with butyric acid-producing bacteria alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BMD and Tb.Th were significantly higher in the LIG + BP group. HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + BP group compared with the LIG group. The expression of OCN and Osterix was significantly higher in the LIG + BP group, while the expression of osteoclast-specific genes (OSCAR, RANK, RANK-L) was significantly lower (P<0.05). ④ Supplementation with butyrate alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BV/TV and Tb.N were significantly higher in the LIG + SB group, and Tb.Sp was significantly lower (P<0.05). HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + SB group compared with the LIG group. The expression of Osterix, OPG, OSCAR, TRAP, and RANK was significantly lower in the LIG + SB group compared with the LIG group (P<0.05). Conclusion Periodontitis disrupts the long bone homeostasis of mice fed a high-fat high-sugar diet, aggravating long bone resorption. Supplementation with butyric acid-producing bacteria or butyrate can effectively alleviate the disruption of long bone homeostasis caused by periodontitis.

Lacking therapeutic targets highlights the crucial roles of chemotherapy and radiotherapy in the clinical management of triple-negative breast cancer (TNBC). To relieve the side effects of the chemoradiotherapy combination regimen, we design and develop a self-assembled micelle nanosystem consisting of perfluorocarbon chain-modified cisplatin prodrug. By incorporating perfluorodecalin, this nanosystem can effectively carry ozone and promote irradiation-derived reactive oxygen species (ROS) production. By leveraging the perfluorocarbon sidechain, the nanosystem exhibits efficient internalization by TNBC cells and effectively escapes from lysosomal entrapment. Under X-ray irradiation, ozone-generated ROS disrupts the intracellular redox balance, thereby facilitating the release of cisplatin in a reduction-responsive manner mediated by reduced glutathione. Moreover, oxygen derived from ozone decomposition enhances the efficacy of radiotherapy by alleviating tumor hypoxia. Notably, the combination of irradiation with ozone-loaded cisplatin prodrug nano system synergistically prompts antitumor efficacy and reduces cellular/systemic toxicity in vitro and in vivo. Furthermore, the combo regimen remodels the tumor microenvironment into an immune-favored state by triggering immunogenic cell death and relieving hypoxia, which provides a promising foundation for a combination regimen of immunotherapy. In conclusion, our nanosystem presents a novel strategy for integrating chemotherapy and radiotherapy to optimize the efficacy and safety of TNBC clinical treatment.

Triple-negative breast cancer (TNBC) remains a refractory subtype of breast cancer due to its resistance to various therapeutic strategies. In this study, we introduce a "brake-release and accelerator-pressing" approach to engineer a microneedle patch embedded with copper-doped Prussian blue nanoparticles (Cu-PB) and the ferroptosis inducer sorafenib (SRF) for raised chemodynamic (CDT)/photothermal (PTT) combination therapy against TNBC. Upon transdermal insertion, the dissolving microneedles swiftly disintegrate and facilitate the release of SRF. Under gentle external light exposure, copper ions (Cu²⁺) and iron ions (Fe³⁺) were liberated from Cu-PB. The direct chelation of Cu²⁺ and the indirect suppression by SRF, collectively attenuate glutathione peroxidase 4 (GPX4) enzymatic function, destabilizing the cellular redox equilibrium (referred to as the "brake-release" strategy). The release of Cu²⁺ and Fe³⁺ ions instigates a Fenton/Fenton-like reaction within tumor cells, further yielding hydroxyl radicals and elevating reactive oxygen species (ROS) concentrations (referred to as the "accelerator-pressing" strategy). This overwhelming ROS accumulation, coupled with the impaired clearance of resultant lipid peroxides (LPO), ultimately triggers a robust ferroptosis cell death response. In summary, this study presents an innovative combinatorial therapeutic strategy based on dual-ferroptosis induction for TNBC, implying a promising therapeutic platform for developing ferroptosis-centered treatments for this aggressive breast cancer subtype.

OBJECTIVES: To investigate pharmacologically active components of Yiqi Zishen Formula (YZF) and their mechanisms for alleviating airway inflammation in mice with chronic obstructive pulmonary disease (COPD). METHODS: Ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry was employed to characterize the chemical components in YZF and YZF-medicated rat serum. A compound-disease target network was constructed based on serum components of YZF to screen the key pathways and targets using enrichment analysis. A mouse model of cigarette smoke-induced COPD was used to evaluate the anti-inflammatory effect of YZF and validate the expression of key proteins in network pharmacology-enriched pathways. Fifty male C57BL/6J mice were randomized equally into control group, COPD model group, high- and low-dose YZF treatment groups, and N-acetylcysteine treatment group. Pulmonary function of the mice was assessed using whole-body plethysmography, and lung histopathology, alveolar structure, and airway remodeling were analyzed using HE staining. The levels of IL-1β, IL-6, and TNF‑α in bronchoalveolar lavage fluid (BALF) were determined with ELISA, and pulmonary expressions of PI3K, Akt, phosphorylated Akt (p-Akt), p65, and phosphorylated p65 (p-p65) were detected using immunohistochemistry. RESULTS: We identified a total of 156 chemical components (including 26 flavonoids or flavonoid glycosides, 27 alkaloids, and 11 saponins) in YZF and 43 prototype components in medicated rat serum. Network pharmacology revealed 704 YZF-related targets and 1199 COPD-associated targets. Integrated analysis suggested that the anti-COPD effects of YZF were associated with the PI3K-Akt signaling pathway. In mouse models of COPD, YZF treatment significantly increased mean alveolar number and peak expiratory flow (P<0.05), reduced mean linear intercept, bronchial wall thickness, lung coefficient, and BALF cytokine levels, and suppressed the expressions of PI3K, Akt, p-Akt, p65, and p-p65 in the lung tissues. CONCLUSIONS YZF alleviates COPD symptoms and airway inflammation in mice possibly by inhibiting the PI3K/Akt/NF‑κB pathway through its multiple components that interact with multiple targets.
Animals ; Pulmonary Disease, Chronic Obstructive/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Signal Transduction/drug effects* ; Male ; Mice, Inbred C57BL ; Mice ; Proto-Oncogene Proteins c-akt/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; NF-kappa B/metabolism* ; Inflammation/drug therapy* ; Rats

Objective To explore the impact of seasonal and temperature variations on fasting blood glucose(FPG)in patients with type 2 diabetes(T2DM).Methods A retrospective analysis was conducted on clinical data of 31 994 T2DM patients managed by community health service center in Minhang District,Shanghai,from January 1,2020 to October 31,2023.The effect of seasonal and temperature variations on FPG levels and blood glucose control was investigated in T2DM patients.Results A total of 72 334 FPG tests were conducted in 31 994 T2DM patients,with an average of 2.26 tests per person.The number of tests conducted in spring,summer,autumn and winter were 15 558,34 018,21 929 and 829,respectively.The FPG levels of T2DM patients were lower in summer and autumn compared to those of winter and spring(P<0.05),while the FPG control rates were higher in summer and autumn compared to those of winter and spring(Bonferroni-corrected,P<0.008).There were no significant differences in FPG levels and blood glucose control rates between spring and winter,or between summer and autumn.The results of the multilevel mixed-effect Logistic model analysis with FPG control status as the dependent variable revealed that the risk of uncontrolled FPG in T2DM patients during winter and spring was 47%(aOR=1.465,95%CI:1.156-1.855)and 51%(aOR=1.510,95%CI:1.384-1.645),respectively,which was higher than that of autumn.The levels of FPG and the incidence of FPG abnormalities reached peak in February and reached the trough in September.Both the FPG levels and the incidence of FPG abnormal exhibited a trend of increasing with the decrease of temperature,and with a lag effect observed.Spearman rank correlation test indicated that there was a negative correlation between FPG levels and incidence of abnormal FPG and the monthly average daily temperature one month lagged(rs=-0.951,P＜0.001 and rs=-0.944,P＜0.001).Conclusion FPG levels of T2DM patients exhibit a pronounced seasonal pattern of elevation during winter and spring,accompanied by a reduction in blood glucose control rates.FPG levels and the prevalence of abnormal FPG exhibit a negative correlation with the monthly average daily temperature,and there is a lag effect.

Objective:To elucidate the genomic characteristics of the immunoglobulin (IG) heavy-chain variable region and light-chain variable region, the expression of subclones, and the prognostic significance in patients with CLL.Methods:Blood and/or bone marrow specimens were gathered from a cohort of 36 patients with CLL diagnosed at Jiangsu Province Hospital from December 2018 to May 2023, including 12 cases of B cell receptor (BCR) stereotyped patients. IG heavy-chain (IGH) and light-chain (IG Kappa [IGK] and IG lambda [IGL]) gene rearrangements were performed using next-generation sequencing (NGS) technology to analyze the characteristics and prognostic value in CLL.Results:NGS detection of IG variable region (IGHV) demonstrated a significant correlation and superior consistency with Sanger sequencing ( r=0.957, P < 0.001). Among the 36 patients, the IGH variant (IGHV) was observed in 9 (25.0%) but not in 27 (75.0%) participants. The incidence of the MYD88 mutation was higher among patients with mutated IGHV [1/27 (3.7%) vs 4/9 (44.4%), P=0.00]. A high incidence of trisomy 12 was observed in the IGHV #8/#8B subset [4/11 (36.4%) vs 1/25 (4.0%), P=0.023], which were more likely to develop Richter transformation [8/11 (72.7%) vs 4/25 (16.0%), P=0.002]. In the patient cohort, 36 individuals (36/36, 100.0%) used the IGK variable, whereas 15 individuals (15/36, 41.7%) employed the IGL variable (IGLV). IGLV3 - 21 reported the highest utilization rate in IGLV (5/15, 33.3%). Remarkably, patients with CLL with IGLV3-21 fragments were exclusively observed in the Binet C stage and Rai Phase Ⅲ-Ⅳ, with an incidence of del (13) (q14) at 60.0% (3/5). The median time to first treatment (TTFT) of patients with or without IGLV3 - 21 fragments was 5.2 (1.1 - 41.5) and 9.9 (0.1 - 94.4) months, respectively. Using the total reads threshold of 2.5%, 4 (4/36, 11.1%) samples were detected to have two IGHV productive clones. The median TTFT and overall survival (OS) time were 2.8 (0.9-72.7) and 12.8 months in patients with one mutated clone and 57.5 (32.0-120.7) and 51.8 months in those with two mutated clones, respectively. The median TTFT and OS time were 10.9 (0.3-94.4) and 6.3 (0.1 - 12.5) months in patients with one unmutated clone and 49.9 (22.2 - 211.1) and 30.0 (9.6 - 50.3) months in those with multiple unmutated clones, respectively ( P>0.05) . Conclusions:Detection of IG gene rearrangements using NGS technology not only facilitates the analysis of the IGHV mutation status, dominant clones, and prognostic value but also contributes to the exploration of IGK/IGL gene rearrangement fragments and the utilization of subclones. Further, it provides information about the poor prognosis of IGLV3 - 21 CLL. The shortened survival of the two unmutated clone groups in the IGHV unmutated group may indicate a poor prognosis.

OBJECTIVE To provide scientific and reasonable recommendations for the identification of Anaerobio-spirillum succiniciproducens in clinical microbiology laboratories by comparing the effectiveness of biochemical methods,matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),and 16S rRNA gene sequencing.METHODS Methodological evaluation was conducted.A strain of A.succiniciprodu-cens was isolated on Mar.27,2023 from a patient with sepsis caused by a dog bite,who was admitted to Guang-dong Nongken Central Hospital,and analyzed for the morphological characteristics,culture methods,and colony features.The identification of the isolated strain was conducted by the automated microbial identification and anal-ysis system(VITEK 2 Compact 2,bioMerieux),MALDI-TOF MS including VITEK MS,MALDI Biotyper,Au-tof ms 1000,and EX-Accuspec,as well as 16S ribosomal RNA(rRNA)gene sequencing.The performance of each method was evaluated.RESULTSA.succiniciproducens was a strictly anaerobic gram-negative spiral bacteri-um,characterized by slow growth,flat and colorless transparent colonies,and negative oxidase and catalase activ-ity.When using VITEK 2 Compact ANC card,it was not successfully recognized;however,through the identifi-cation by VITEK MS,MALDI Biotyper,Autof ms 1000 and EX-Accuspec,it was all confirmed as A.succinicip-roducens.Additionally,16S rRNA gene sequencing technology also identified it as A.succiniciproducens.CONCLUSIONS Current automated biochemical identification systems are unable to accurately identify A.succi-niciproducens.MALDI-TOF MS and 16S rRNA gene sequencing techniques can reliably confirm A.succinicipro-ducens.Clinical microbiologists should select the appropriate identification method based on their available re-sources.

Objective:To investigate the clinicopathological features, diagnosis, and prognosis of aggressive natural killer-cell leukemia (ANKL).Methods:A retrospective analysis was conducted on 27 ANKL patients treated at the First Affiliated Hospital of Nanjing Medical University from 2014 to 2024. Their clinical data, histomorphology, and immunophenotype were reviewed. Kaplan-Meier analysis was used to evaluate the overall survival (OS), and COX regression analysis was performed to identify prognostic factors affecting OS.Results:Among the 27 patients, 18 were male and 9 were female, with a male-to-female ratio of 2∶1. The age ranged from 15-75 years, with a median age of 42.0 (28.5, 54.5) years. Fever and splenomegaly were the most common signs and symptoms. Most patients presented with pancytopenia, coagulation abnormalities, and liver dysfunction; and all patients had elevated EBV loads. Microscopically, 16 cases showed marked to hypercellular bone marrow proliferation, with predominant interstitial infiltration (15 cases, 55.6%), followed by sinusoidal infiltration (3 cases), diffuse infiltration (6 cases, 22.2%), mixed infiltration (interstitial and focal, 3 cases, 11.1%), focal infiltration (2 cases, 7.4%), and nodular infiltration (1 case, 3.7%). The proportion of tumor cells among nucleated cells ranged from 2% to 80%, with a median of 30%. The tumor cells displayed variable morphology. Hemophagocytosis was observed in 23 cases. Immunohistochemistry revealed that all cases expressed CD56, with mostly expressing cytotoxic molecules (granzyme B, TIA-1). The Ki-67 proliferative index ranged from 50% to 90%. CD56-EBER dual staining showed that NK cells were the primary targets of the virus. Reticulin staining showed increased fibrosis. By flow cytometry, all cases were positive for CD2 but negative for surface CD3 (sCD3), CD4, CD5 and CD57. Among them, 21 cases (95.5%) exhibited a typical phenotype of strong CD56 expression (CD56str+) with CD16 negativity (CD16-), while only one case (4.5%) showed CD16 positivity (CD16+) with dim CD56 expression (CD56dim). In killer-cell immunoglobulin-like receptor (KIR) analysis, 6 out of 17 patients (6/17) demonstrated monoclonal expression, including CD158a (4/6), CD158i (1/6), and CD158e (1/6); the remaining 11 cases (11/17) showed complete absence of KIR expression. All tested cases (17/17) were negative for T-cell receptor (TCR) protein expression. Follow-up period was from 257 days, 1 patient was lost to follow-up, and the remaining 26 patients died. Kaplan-Meier analysis revealed that OS was significantly longer in patients who received chemotherapy compared to those who did not ( P<0.05). Univariate Cox proportional hazards model analysis indicated that age, bone marrow proliferation, proportion of tumor cells among nucleated cells, absolute neutrophil count, platelet count, and triglycerides and bilirubin levels significantly affected OS ( P<0.05). Multivariate COX regression analysis identified triglycerides and bilirubin levels as independent prognostic factors for OS. Conclusion:Aggressive natural killer-cell leukemia is a rare lymphoid malignancy with very poor prognosis. Tumor cells exhibit significant morphological variation, and bone marrow infiltration patterns are diverse. Accurate recognition, early diagnosis, and timely chemotherapy are critical to improving the prognosis of patients with ANKL.

Patients with leukemic non-nodal mantle cell lymphoma (nnMCL) typically exhibit an indolent clinical course, and when asymptomatic and without treatment indications, a watchful observation follow-up can be adopted. This article presents a retrospective summary of a case of nnMCL, misdiagnosed as chronic lymphocytic leukemia (CLL) and carrying a TP53 mutation, along with a literature review. The case highlights the importance of differential diagnosis between nnMCL and CLL, and suggests that for nnMCL patients, the presence of high-risk biological markers such as TP53 mutations does not necessarily indicate an immediate need for treatment; rather, a strict watch-and-wait strategy may be a more appropriate option.

Objective To investigate the labeling of dialysis-related information in the instructions of drugs for systemic use in patients with end-stage renal disease undergoing hemodialysis,and to provide reference for further standardization and im-provement of drug instructions.Methods Collected the information on pharmacokinetics and drug dose adjustment in hemodi-alysis patients with end-stage renal disease from the instructions of chemicals and biologics used systemically in XuanWu hospital's formulary from January to March 2023.Classified and compared the labeling rates of the corresponding information of the originally developed drugs and generic drugs,imported drugs and domestic drugs;Drugs eliminated by non-renal route,or with mo-lecular weight≥5 000,or binding rate of plasma protein ≥ 60％,or drug distribution volume＞360L were classified as"non-dialysis group",and other drugs were classified as"dialysis group",compared the labeling rate of corresponding information of drugs in"dialysis group"and"non-dialysis group";and compared the labeling rate and content with relevant information in Lexicomp and Micromedex.Results Among the 930 drug instructions,the labeling rates of drug dialysis clearance,drug adjustment,and ex-plicit drug adjustment plan were 16.67％,25.16％,and 24.52％.There was no significant difference in the labeling rate of dialysis-related information between original and generic drugs,and imported and domestic drugs.There was a significant difference in the labeling rate of dialysis-related information between the"dialyzable group"and the"non-dialyzable group"(P＜0.01).The differ-ence in the labeling rate of dialysis related information between the investigated drug instructions and the corresponding drugs in Lexicomp or Micromedex was statistically significant(P＜0.01).Conclusion Attention should be paid to the lack of informa-tion and unclear labeling of hemodialysis patients with end-stage renal disease in the drug instructions by relevant departments.