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    Frontier review
    Applications and progress of AI in mechanistic research and clinical diagnosis of oral genetic and rare diseases
    Shen Jingting, Liu Zhixu, Wang Xudong
    2026, 46 (7):  829-838. 
    doi: 10.3969/j.issn.1674-8115.2026.07.001

    Abstract ( 20 )   HTML ( 0 )   PDF (1735KB) ( 3 )  

    Oral genetic and rare diseases are characterized by low prevalence, marked phenotypic heterogeneity, and complex pathogenic mechanisms. They often present with severe clinical manifestations, pose significant diagnostic challenges, and lack effective therapeutic options. With the rapid advancement of multi‑omics sequencing and medical imaging technologies, large‑scale molecular and clinical data have provided new perspectives for understanding these disorders. Artificial intelligence (AI), with its powerful capabilities in pattern recognition and modeling complex relationships, has achieved significant progress in both fundamental research and clinical diagnosis and treatment related to oral genetic and rare diseases. In studies of pathogenic mechanisms, AI enables the explanation of genomic, transcriptomic, proteomic, and microbiome data, facilitates the identification of novel molecular biomarkers, and supports the construction of disease prediction models, thereby advancing the elucidation of complex etiologies. In clinical diagnosis, AI significantly enhances the automation and accuracy of oral imaging analysis, enabling intelligent diagnosis of dental caries, tooth developmental anomalies, salivary gland diseases, fibrous dysplasia, and various craniofacial malformations. AI‑based tools also provide decision support for surgical treatment planning. Despite these advances, challenges remain, including data scarcity, limited model interpretability, and ethical and regulatory concerns. This paper aims to provide a comprehensive review of the latest advancements in the application of AI in the field of oral genetic and rare diseases, systematically examining the progress, challenges, and prospects in deepening the understanding of disease mechanisms and accelerating clinical diagnosis, with the goal of ultimately promoting the overall advancement of precision medicine for oral genetic diseases and improving the accessibility of high-quality healthcare resources.

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    Research progress in adaptive survival and pathogenic mechanisms of toxin-antitoxin systems in Staphylococcus aureus
    Xie Xinni, Lü Yan, Li Min, Wang Yanan
    2026, 46 (7):  839-846. 
    doi: 10.3969/j.issn.1674-8115.2026.07.002

    Abstract ( 14 )   HTML ( 0 )   PDF (1886KB) ( 2 )  

    Toxin-antitoxin systems (TAS) are widely distributed in bacteria and archaea, typically consisting of a toxin that inhibits bacterial growth and a cognate antitoxin that neutralizes its toxicity. These systems have been recognized as key regulators of bacterial growth, metabolism, and pathogenesis. Staphylococcus aureus (S. aureus) is an important clinical pathogen that can cause a wide range of diseases, ranging from skin and soft tissue infections to sepsis and endocarditis. Studies have shown that TAS play important roles in stress resistance, persister cell formation, and biofilm development in S. aureus. Elucidating the underlying molecular mechanisms is essential for understanding the pathogenic characteristics of S. aureus and for developing novel antibacterial strategies. Here, This review summarizes the classification and structural features of TAS in S. aureus and discuss their functions in adaptive survival and virulence regulation. It focuses on how, under nutrient limitation or antibiotic pressure, TAS interfere with DNA replication and protein synthesis to induce a metabolically dormant state, thereby contributing to phenotypic tolerance and persister cell formation. This review further describes how TAS modulate virulence factor expression through global regulatory networks and how certain TAS-encoded toxins directly damage eukaryotic membranes to promote bacterial invasion and dissemination. Additionally, it highlights the contribution of the type Ⅶ secretion system to toxin export, which provides a competitive advantage to S. aureus within polymicrobial communities. This review systematically summarizes the structural characteristics and functional modes of different types of TAS in S. aureus, aiming to deepen the understanding of its pathogenic mechanisms and to provide theoretical references and research directions for the development of novel TAS-targeted antibacterial strategies.

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    Review of indication stratification and clinical strategies for endoscopic ultrasound-guided pancreatic duct drainage
    Chang Yuxin, Li Baiwen, Ni Jianbo
    2026, 46 (7):  847-856. 
    doi: 10.3969/j.issn.1674-8115.2026.07.003

    Abstract ( 7 )   HTML ( 3 )   PDF (11360KB) ( 6 )  

    Endoscopic ultrasound-guided pancreatic duct drainage (EUS-PD/EUS-PDD) represents a major advancement in therapeutic endoscopy for pancreatic duct disorders. It is primarily indicated for patients with symptomatic pancreatic duct obstruction in whom endoscopic retrograde pancreatography (ERCP) has failed or is not feasible. The procedure involves transmural puncture of the main pancreatic duct via the stomach, duodenum, or postsurgical jejunal limb, followed by guidewire placement, tract dilation, and stent insertion to achieve ductal decompression and restoration of pancreatic juice drainage. Systematic reviews and meta-analyses have reported an overall technical success rate of 84.8%, a clinical success rate of 89.2%, and an overall adverse event rate of 18.1%, indicating substantial efficacy with non-negligible risks. The common indications for EUS-PD include pancreatic duct disruption due to pancreatic trauma, pancreatic duct injury associated with pancreatic surgery, pancreatic duct disruption secondary to severe acute pancreatitis, and surgically altered anatomy. Complications mainly include post-procedural pancreatitis, bleeding, perforation, and stent-related dysfunction. This review constructs an evidence chain covering indication stratification, procedural approaches, key techniques, complication management, long-term follow-up, and reintervention strategies, aiming to provide a reproducible clinical decision-making framework for the implementation of EUS-PD in endoscopic centers in China.

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    Basic research
    Effect of suppressor of cytokine signaling 3 on osteogenic differentiation of mouse calvarial osteoblast precursors
    Cui Yiwen, Sun Siyuan, Huang Zihan, Dai Qinggang, Jiang Lingyong
    2026, 46 (7):  857-867. 
    doi: 10.3969/j.issn.1674-8115.2026.07.004

    Abstract ( 6 )   HTML ( 0 )   PDF (69987KB) ( 4 )  

    Objective ·To construct a stable suppressor of cytokine signaling 3 (Socs3)-knockdown MC3T3-E1 mouse calvarial osteoblast precursor cell line and to investigate the role of SOCS3 in the osteogenic differentiation of calvarial osteoblast precursors. Methods ·The expression levels of SOCS3 in MC3T3-E1 cells after osteogenic induction were detected by real-time quantitative PCR (RT-qPCR) and Western blotting. Two pairs of short hairpin RNAs (shRNAs) specifically targeting the Socs3 gene were designed, and lentiviral vectors for Socs3 knockdown were constructed. MC3T3‑E1 cells were then infected with the packaged lentiviruses, followed by puromycin screening to obtain MC3T3‑E1 cell lines with stable Socs3 knockdown. Knockdown efficiency was verified by RT-qPCR and Western blotting. The effect of SOCS3 on the proliferation capacity of MC3T3-E1 cells was assessed using a CCK-8 assay. After osteogenic differentiation for 7 d, alkaline phosphatase (ALP) staining and RT‑qPCR detection of osteogenesis‑related genes were performed. After 14 d of osteogenic differentiation, alizarin red S (ARS) staining was performed to assess the osteogenic differentiation capacity of the stable knockdown cell lines. Results ·RT-qPCR and Western blotting showed that both Socs3 mRNA and SOCS3 protein levels were elevated at 4 d and 7 d after osteogenic induction compared with those before induction, and the levels at 7 d were higher than those at 4 d (both P<0.001). In MC3T3-E1 stable cell lines infected with shSocs3-1 and shSocs3-2 lentivirus, the expression levels of Socs3 mRNA and SOCS3 protein were significantly inhibited (P<0.001), indicating the successful establishment of stable Socs3-knockdown MC3T3-E1 cell lines. The CCK-8 assay showed that Socs3 knockdown inhibited the proliferation capacity of MC3T3-E1 cells at 1 d, 4 d, and 7 d of culture (P<0.001). ALP staining demonstrated reduced ALP activity in the Socs3-knockdown MC3T3-E1 cell lines after 7 d of osteogenic induction (P<0.001). ARS staining indicated that Socs3 knockdown inhibited mineralized nodule formation in MC3T3-E1 cells (P<0.001). RT-qPCR detection of osteogenic marker genes indicated that the expression levels of runt-related transcription factor 2 (Runx2), alkaline phosphatase (Alp), osterix (Osx), α1 type Ⅰ collagen (Col1a1), secreted phosphoprotein 1 (Spp1), and bone γ-carboxyglutamate protein (Bglap) were significantly reduced in the Socs3-knockdown group (all P<0.001). Conclusion ·SOCS3 expression increases during the osteogenic differentiation of MC3T3-E1 cells. Knockdown of Socs3 can inhibit both the proliferation and osteogenic differentiation capacities of MC3T3-E1 cells.

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    Promotion of tendon healing by rosmarinic acid-based hydrogel
    Wang Yuan, Wu Rongpu, Wang Fei, Liu Shen
    2026, 46 (7):  868-874. 
    doi: 10.3969/j.issn.1674-8115.2026.07.005

    Abstract ( 7 )   HTML ( 0 )   PDF (15503KB) ( 4 )  

    Objective ·To construct a rosmarinic acid (RA)-loaded hydrogel and investigate its effects on tissue repair and functional recovery after Achilles tendon injury. Methods ·Phenylboronic acid-modified carboxymethyl chitosan (CMCS-PBA) was synthesized via an amidation reaction between phenylboronic acid and carboxymethyl chitosan, and the molecular structure of CMCS-PBA was confirmed by 1H nuclear magnetic resonance (1H-NMR) spectroscopy. CMCS-PBA/RA hydrogel was prepared by mixing CMCS-PBA with RA at a volume ratio of 10∶1. CMCS-PBA/PVA hydrogel composed of CMCS-PBA and polyvinyl alcohol (PVA) served as the positive control. The rheological property, self-healing ability, and mechanical performance of the hydrogel were systematically characterized with a rheometer. Rat fibroblast cell line 208F was used to evaluate the biocompatibility of the hydrogels via live/dead cell staining. Eighteen rats with established Achilles tendon transection models were randomly assigned into three groups (n=6 per group): the control group, the CMCS-PBA/RA group, and the CMCS-PBA/PVA group. In the CMCS-PBA/RA and CMCS-PBA/PVA groups, the corresponding hydrogels were locally injected into the injured tendons immediately after model establishment to completely cover the injury site, while no hydrogel intervention was administered in the control group. All rats were sacrificed 3 weeks postoperatively. Hematoxylin-eosin (HE) staining and Masson staining were performed to evaluate tendon healing, and gait analysis was conducted to assess functional recovery. Major visceral organs were harvested for HE staining to further verify the in vivo biocompatibility of the prepared hydrogels. Results ·1H-NMR results confirmed the successful grafting of PBA onto the CMCS molecular chains. Rheological characterization demonstrated that the prepared hydrogels possessed excellent shear-thinning behavior, self-healing ability, and favorable injectability. Live/dead staining and organ HE staining verified the favorable biocompatibility of the hydrogels. Compared with the control and CMCS-PBA/PVA groups, the CMCS-PBA/RA group exhibited significantly lower tendon healing scores (both P<0.05), indicating superior tendon healing. Gait analysis showed a significantly larger footprint area in the CMCS-PBA/RA group (both P<0.05), which reflected better functional recovery. Conclusion ·A novel CMCS-PBA/RA composite hydrogel was successfully prepared. The hydrogel possesses excellent injectability, self-healing capability, and biocompatibility, and effectively promotes Achilles tendon tissue repair and motor functional recovery.

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    Mechanism of WNT5A in aggravating the ischemia-reperfusion injury-induced acute kidney injury to chronic kidney disease transition by promoting FOXP1
    Feng Haoran, Liu Junjun, Wang Kaichun, Wang Niansong, Gu Sijie, Fan Ying
    2026, 46 (7):  875-885. 
    doi: 10.3969/j.issn.1674-8115.2026.07.006

    Abstract ( 13 )   HTML ( 0 )   PDF (41797KB) ( 12 )  

    Objective ·To investigate the mechanism by which wingless-type MMTV integration site family member 5A (WNT5A) promotes the transition from ischemia-reperfusion injury (IRI)-induced acute kidney injury (AKI) to chronic kidney disease (CKD). Methods ·An IRI-induced AKI-to-CKD transition model was established in wild-type (WT) mice. Renal histopathological injury was assessed by hematoxylin-eosin (H-E) staining on the 3rd and the 14th day after IRI. Fibrosis was evaluated by Masson staining and immunohistochemistry for collagen type Ⅰ α1 chain (COL1A1). Wnt5a expression was quantified by RNA sequencing (RNA-seq), real-time quantitative PCR (qPCR), and Western blotting. Wnt5a heterozygous knockout (Wnt5a+/- ) mice were used to establish the IRI-induced AKI-to-CKD transition model. Renal function was evaluated by serum creatinine (Scr) and blood urea nitrogen (BUN), and renal injury and fibrosis were assessed as described above. In vitro, a transforming growth factor-β (TGF-β)-induced fibrotic model was established in HK-2 cells (a human renal cortex proximal tubular epithelial cell line) with WNT5A overexpression. Differentially expressed genes identified by RNA-seq were subjected to Gene Ontology (GO) biological process enrichment analysis to identify key targets. Regulation of forkhead box protein P1 (FOXP1) by WNT5A was validated by qPCR and Western blotting. FOXP1 was overexpressed, or silenced in the context of WNT5A overexpression in HK-2 cells. Expression levels of notch receptor 2 (NOTCH2) pathway components, including NOTCH2, hairy/enhancer-of-split related with YRPW motif 1(HEY1), and hairy and enhancer of split 1 (HES1), as well as fibrosis markers, including COL1A1 and vimentin (VIM), were analyzed by qPCR and Western blotting. Results ·Wnt5a expression was increased in the renal tissue on the 3rd day after IRI, accompanied by marked tubular dilation and inflammatory cell infiltration. On the 14th day, Wnt5a expression was further increased, along with significant upregulation of fibrotic markers. Compared with WT mice, Wnt5a+/- mice exhibited reduced Scr and BUN levels on the 14th day after IRI, indicating improved renal function. FOXP1 expression was decreased, and tubular injury and renal fibrosis were attenuated in Wnt5a+/- mice. Mechanistically, WNT5A overexpression markedly upregulated FOXP1 in TGF-β-treated HK-2 cells. RNA-seq and GO analysis indicated significant enrichment of fibrosis-related processes and activation of the NOTCH signaling pathway. FOXP1 overexpression increased the mRNA and protein levels of NOTCH2 and its downstream targets HEY1 and HES1, along with elevated COL1A1 and VIM expression. In contrast, silencing FOXP1 in WNT5A-overexpressing cells reversed the upregulation of NOTCH2 pathway components and fibrosis markers. Conclusion ·WNT5A upregulates the transcription factor FOXP1, which activates the NOTCH2 signaling pathway, thereby accelerating the IRI-induced AKI-to-CKD transition, and promoting renal fibrosis progression.

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    Anti-inflammatory and antioxidant effects of tannic acid-based ROS-responsive hydrogel in macrophage inflammation
    Wu Jialong, Chen Xuzhuo, Huang Enkui, Yin Xiuyuan, Xu Qianhe, Liu Peinian, Zhang Shanyong
    2026, 46 (7):  886-895. 
    doi: 10.3969/j.issn.1674-8115.2026.07.007

    Abstract ( 7 )   HTML ( 0 )   PDF (38478KB) ( 1 )  

    Objective ·To develop a reactive oxygen species (ROS)-responsive hydrogel based on tannic acid (TA), and to evaluate its anti-inflammatory and antioxidant effects in an lipopolysaccharide (LPS)-induced RAW264.7 macrophage inflammation model. Methods ·Phenylboronic acid-modified hyaluronic acid (HP) was complexed with TA to form the HP-TA hydrogel, and its ROS-responsive properties were assessed by using a hydrogen peroxide consumption assay. The biocompatibility of HP and TA in RAW264.7 macrophages was evaluated by cell counting kit-8 (CCK-8) assay, cell cycle assay, and live/dead staining. An LPS-induced inflammation model of RAW264.7 macrophages was established and treated with the HP-TA hydrogel. The mRNA and protein expression levels of pro-inflammatory genes, including inducible nitric oxide synthase (Inos) and tumor necrosis factor-α (Tnf-α), as well as antioxidant genes, including NAD(P)H: quinone oxidoreductase 1 (Nqo1) and heme oxygenase-1 (Ho-1), were analyzed by quantitative real-time PCR (qPCR) and Western blotting, respectively. A 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe combined with flow cytometry and nitric oxide content detection was performed to analyze the total ROS-scavenging capacity of the hydrogel. Results ·The HP-TA hydrogel was successfully synthesized and confirmed to be ROS-responsive. At safe concentrations (200 μg/mL for HP and 25 μg/mL for TA), both materials exhibited good biocompatibility with RAW264.7 macrophages. The HP-TA hydrogel significantly inhibited the mRNA and protein expression of the pro-inflammatory genes Inos and Tnf-α, and promoted the mRNA and protein expression of the antioxidant genes Nqo1 and Ho-1 in inflammatory macrophages (all P<0.05). In addition, the HP-TA hydrogel significantly reduced the total intracellular ROS level (P<0.001). Conclusion ·The HP-TA hydrogel can effectively suppress LPS-induced inflammatory responses in macrophages, exert antioxidant effects by scavenging ROS, and upregulate the expression of antioxidant pathway proteins. This study reveals the potential of the HP-TA hydrogel as a ROS-responsive intelligent drug delivery system.

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    Comparison of inner hair cell transduction by different serotypes of adeno-associated virus via cisterna magna injection in mice
    An Yiran, Feng Baoyi, Sun Yilin, Wu Hao, Tao Yong
    2026, 46 (7):  896-905. 
    doi: 10.3969/j.issn.1674-8115.2026.07.008

    Abstract ( 8 )   HTML ( 0 )   PDF (128976KB) ( 1 )  

    Objective ·To evaluate the safety of cisterna magna-delivered adeno-associated virus (AAV) of different serotypes carrying the human cytomegalovirus immediate-early promoter (CMV) in the auditory and central nervous systems, as well as their transduction differences in inner hair cells (IHCs) and the central nervous system. Methods ·Eight AAV serotypes (AAV1, AAV2, AAV8, AAV9, AAV-rh10, AAV-DJ, AAV-PHP.B, and AAV-PHP.eB) carrying enhanced green fluorescent protein (EGFP) were injected into the cerebrospinal fluid of 4-week-old wild-type mice via cisterna magna. Auditory brainstem response (ABR) was measured to evaluate changes in auditory thresholds before injection and 2 weeks after injection. At 2 weeks post-injection, cochleae were collected and subjected to immunofluorescence staining to compare the transduction efficiency and number of cochlear IHCs among the eight AAV-EGFP serotypes. The EGFP fluorescence signal intensity of the three serotypes with higher transduction efficiency was further analyzed in IHCs. Fluorescent DNA dye staining on frozen brain sections was used to compare the central nervous system transduction characteristics of the three serotypes. Open-field test was performed to assess the effects of cisterna magna injection of three AAV-EGFP serotypes on central nervous system safety in mice. Results ·ABR tests showed no significant differences in auditory thresholds at all tested frequencies before and 2 weeks after cisterna magna injection. Immunofluorescence staining demonstrated that all eight AAV-EGFP serotypes successfully transduced IHCs in the 32 kHz cochlear region at 2 weeks after in jection. Among them, AAV2-CMV-EGFP [(83.33±14.79)%], AAV-DJ-CMV-EGFP [(82.52±5.18)%], and AAV-PHP.B-CMV-EGFP [(96.61±1.87)%] exhibited relatively high transduction efficiency in IHCs at 32 kHz, and no significant intergroup difference was observed in the number of IHCs after injection. Analysis of EGFP fluorescence intensity in the three serotypes with the highest transduction efficiency revealed stronger fluorescence signals in IHCs transduced by AAV-PHP.B-CMV-EGFP and AAV2-CMV-EGFP at the 32 kHz region. Staining of frozen brain sections revealed distinct transduction patterns of the three AAV-EGFP serotypes in the central nervous system following cisterna magna injection. AAV2-CMV-EGFP exhibited relatively low transduction levels; AAV-DJ-CMV-EGFP showed detectable transduction in the cerebellar cortex, dorsal medulla, and periventricular regions; AAV-PHP.B-CMV-EGFP showed transduction in the molecular and granular layers of the cerebellar cortex and periventricular regions. Open-field test results indicated that spontaneous locomotor activity of mice remained normal following cisterna magna injection of the three tested viral serotypes. Conclusion ·Cisterna magna injection of different AAV-EGFP serotypes causes neither hearing impairment nor IHCs injury in mice. Different AAV-EGFP serotypes exhibit distinct transduction efficiencies in the cochlea and central nervous system.

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    4T1 cell membrane-biomimetic nanoprobe based on glutathione depletion for inducing tumor ferroptosis and MR imaging
    Ding Xinyi, Zhu Yi, Wang Jingyi, Deng Jiali, Wang Zhongling
    2026, 46 (7):  906-915. 
    doi: 10.3969/j.issn.1674-8115.2026.07.009

    Abstract ( 5 )   HTML ( 0 )   PDF (41122KB) ( 2 )  

    Objective ·To develop a biomimetic membrane, glutathione (GSH)-responsive manganese-based theranostic nanoprobe, and investigate its biomembrane-targeting capability, efficacy in GSH-depletion-mediated ferroptosis induction in triple-negative breast cancer (TNBC), and real-time monitoring of MR signal activation. Methods ·A silica (SiO2) nanoparticle core was synthesized via the sol-gel method, followed by deposition of manganese dioxide (MnO2) onto its surface through redox co-precipitation, and then coating with 4T1 breast cancer cell membranes using an extrusion technique to yield the biomimetic nanoprobe SiO2@MnO2@Membrane (SMM). The morphology, particle size, and Zeta potential were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). Membrane protein retention on the nanoparticle surface was confirmed by Coomassie brilliant blue staining, and characteristic elements was analyzed by X-ray photoelectron spectroscopy (XPS). The GSH-responsive T1 MR signal activation was evaluated in vitro. Reactive oxygen species (ROS) generation and GSH depletion in a cell-free system were assessed using methylene blue and 5, 5′-dithiobis (2-nitrobenzoic acid) (DTNB) assays, respectively. The cytotoxic effect of the nanoprobe on 4T1 cells was determined by the MTT assay. Intracellular ROS generation and lipid peroxide (LPO) accumulation were detected using 2′,7′-dichlorodihydrofluoresce in diacetate (DCFH-DA) and BODIPY 581/591 C11 probes. A murine subcutaneous TNBC model was established to dynamically monitor T1-weighted MR signal activation of the nanoprobe at tumor sites. Combined with the detection of ferroptosis-related markers, including LPO and ROS, as well as Ki-67 and terminal-deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) staining in tumor tissues, the ferroptosis-mediated antitumor efficacy of SMM was evaluated. Results ·TEM revealed that the prepared SMM nanoprobe exhibited a regular spherical structure. Particle size analysis showed that after cell membrane coating, the Zeta potential shifted from (+3.2±1.0) mV to (-22.0±2.0) mV, and the hydrodynamic diameter increased from 68 nm to 78 nm, while the polydisper sity index (PDI) remained at a low level, indicating a uniform and stable dispersion. Coomassie brilliant blue staining confirmed effective biomimetic membrane coating. XPS detected characteristic signals of Mn and Si elements, and these results collectively confirmed the successful synthesis of the SMM nanoprobe. MR results showed that SMM significantly activated the T1 signal in the presence of GSH, with relaxivity (r1). increasing significantly from 0.591 to 6.875 (mmol/L)-1·s-1. At the cellular level, T1 signal activation exhibited a time-dependent enhancement. Compared to the control groups, DCFH-DA and BODIPY 581/591 C11 staining indicated substantial intracellular accumulation of ROS and LPO in the SMM group, accompanied by a significant reduction in cell viability (28.35%±4.91% survival). In vivo experiments showed that after intravenous injection of SMM,the T1 signal intensity in tumor regions reached a peak value of (1.23±0.14) s-1 at 4 h, indicating that the nanoprobe could specifically accumulate in TNBC tumor tissues and activate MR T1 signals. Tumor tissue analysis revealed significantly elevated ROS and LPO levels in the SMM group, accompanied by decreased Ki-67 expression and increased TUNEL-positive signals, further confirming its antitumor effect through ferroptosis induction. Conclusion ·A GSH-depleting 4T1 cell membrane-biomimetic nanoprobe (SMM) was successfully developed, which can specifically activate T1 MR signals at TNBC tumor sites and synergistically induce ferroptosis through targeted accumulation, significantly enhancing antitumor efficacy.

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    Engineering and structural analysis of heterologous protein loading based on an effector of the rearrangement hotspot family in the type Ⅵ secretion system
    Pan Yue, Huang Jing
    2026, 46 (7):  916-927. 
    doi: 10.3969/j.issn.1674-8115.2026.07.010

    Abstract ( 15 )   HTML ( 0 )   PDF (36568KB) ( 3 )  

    Objective ·To systematically evaluate the loading capacity of the barrel-shaped structure of the rearrangement hotspot (RHS) effector type Ⅵ secretion system effector I (TSEI) from Aeromonas dhakensis (A. dhakensis) within the type Ⅵ secretion system (T6SS) for heterologous proteins and explore its feasibility as an engineered intercellular protein delivery module from a structural perspective. Methods ·The genes encoding the A. dhakensis T6SS-related proteins valine-glycine repeat protein G (VGRG), type Ⅵ secretion system effector chaperone I (TECI), and the engineered TSEI effector generated by heterologous protein replacement were individually cloned into vectors suitable for expression in Escherichia coli (E. coli).The component proteins were co-expressed in E. coli via chemical competent transformation. Stable expression of the complexes was achieved by regulating the induction conditions. Following cell lysis, the two target complexes were initially purified separately using tag-mediated affinity chromatography, and then were further separated via size-exclusion chromatography to improve sample homogeneity and integrity. A glycerol density gradient centrifugation step was introduced to remove aggregates and enrich fully assembled complexes. The composition and optimal state of the complexes were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. High-purity recombinant samples were subjected to cryo-electron microscopy (cryo-EM) data collection. Single-particle analysis, including particle picking, two-dimensional classification, and three-dimensional reconstruction, was carried out to obtain 3D structural models. UCSF Chimera and other visualization software were used to fit known or predicted protein structures into the electron density maps, enabling inference of the relative spatial arrangement of components and providing structural evidence for the assembly state of the engineered TSEI within T6SS-related complexes. Results ·Through co-expression in E. coli and sequential purification using affinity chromatography, size-exclusion chromatography, and glycerol density gradient centrifugation, two recombinant effector module complexes with well-defined compositions were successfully acquired, as confirmed by SDS-PAGE and Western blotting analyses. These results indicated that the RHS barrel structure exhibited good tolerance for heterologous protein loading. Cryo-EM density maps showed that, the overall conformation of the complex remained stable after replacement with monomeric cherry fluorescent protein (MCHERRY), with additional density observed inside the RHS barrel, suggesting successful loading. In contrast, replacement with CRISPR-associated endonuclease Φ (CASΦ) likely reduced the stability of the barrel scaffold and restricted the formation of a fully encapsulated structure. These findings demonstrated that the RHS barrel possesses the capacity to accommodate heterologous proteins within a certain range and revealed its structural adaptation boundaries, providing structural insights for the development of T6SS-based engineered protein delivery platforms. Conclusion ·Based on recombinant expression and cryo-EM structural analysis, this study systematically validated the feasibility of the RHS barrel of the A. dhakensis T6SS effector TSEI for delivering heterologous proteins, offering experimental support for T6SS-based targeted protein transport and functional engineering.

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    Clinical research
    Analysis of clinical characteristics of three patients with arrhythmogenic cardiomyopathy carrying novel pathogenic PKP2 variants
    Ni Luyan, Wu Chen, Tao Zhengyu, Wang Xiaoning, Zhang Zhixuan, Dong Jiawei, Jiang Meng
    2026, 46 (7):  928-937. 
    doi: 10.3969/j.issn.1674-8115.2026.07.011

    Abstract ( 7 )   HTML ( 0 )   PDF (8669KB) ( 2 )  

    Objective ·To report newly discovered pathogenic variants of the PKP2 gene in the Chinese population and analyze their association with the clinical phenotype of arrhythmogenic cardiomyopathy (ACM). Methods ·Clinical data, including genetic test results and imaging features, were collected from 251 Chinese patients with unexplained cardiomyopathy. All detected rare variants were analyzed by whole-exome sequencing and confirmed by Sanger sequencing. The clinical phenotypes of patients carrying PKP2 variants were analyzed. Results ·Eight patients were diagnosed with arrhythmogenic cardiomyopathy, among whom 3 (37.5%) carried novel PKP2 variants in the Chinese population. Among the three variants, the missense variant c.1256T>C (p.Leu419Ser) was associated with left ventricular involvement, the missense variant c.2264T>C (p.Leu755Ser) was associated with right ventricular involvement, and the splice-site variant c.2167+1G>C was associated with biventricular involvement and increased susceptibility to ventricular tachycardia. All three patients presented with arrhythmias, and the mean age at disease onset was (23.3±10.5) years. Conclusion ·The novel pathogenic PKP2 variants identified in Chinese population expand the variant spectrum of the PKP2 gene. Key differences in ventricular involvement patterns (univentricular or biventricular) and susceptibility to ventricular tachycardia are revealed between gain-of-function (missense variants) and loss-of-function (splicing variant) mutations of the PKP2 gene.

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    Screening of specific metabolic biomarkers for brucellosis and construction of a differential diagnostic model based on serum metabolomics
    Zhou Jinping, He Xiaoyan, Song Wen, Liu Yumei, Huang Lin, Ma Xiumin
    2026, 46 (7):  938-945. 
    doi: 10.3969/j.issn.1674-8115.2026.07.012

    Abstract ( 7 )   HTML ( 0 )   PDF (1967KB) ( 3 )  

    Objective ·To analyze the correlations between differential metabolic features and clinical indicators in patients with brucellosis and rheumatoid arthritis, and to identify biomarkers with potential for differential diagnosis. Methods ·Clinical data were collected from 70 patients with brucellosis and 75 patients with rheumatoid arthritis, and baseline characteristics and clinical laboratory indicators were compared between the two groups. Serum metabolomic profiles were obtained using nanoparticle-enhanced laser desorption/ionization mass spectrometry (NPELDI-MS) to identity differentially expressed metabolites between the groups. Furthermore, Spearman′s rank correlation analysis was performed to identify brucellosis-specific metabolites significantly correlated with clinical indicators in the brucellosis group, followed by pathway enrichment analysis. A neural network-based diagnostic model was constructed based on these metabolites. Results ·Compared with the rheumatoid arthritis group, the brucellosis group demonstrated significantly lower erythrocyte sedimentation rate, neutrophil count, and serum creatinine levels (all P<0.05). In contrast, red blood cell count, hemoglobin level, and liver function-related indicators (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total protein) were all significantly higher in the brucellosis group (all P<0.05). Among the 273 metabolic features obtained by NPELDI-MS, 133 showed significant differences between the two groups. Correlation analysis of these 133 differential metabolic features with the significantly different clinical indicators identified nine brucellosis-specific metabolites: taurine, pyroglutamic acid, uric acid, L-aspartic acid, glutamine, phenylalanine, urocanic acid, dihydroxyacetone phosphate, and pyrophosphate. These metabolites showed significant negative correlations with liver function-related indicators (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase) and the inflammatory indicator (neutrophil count) (all P<0.05). Pathway enrichment analysis demonstrated that these metabolites were significantly enriched in several key metabolic pathways, including histidine metabolism; alanine, aspartate, and glutamate metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; phenylalanine metabolism; and taurine and hypotaurine metabolism. The diagnostic model constructed based on the combination of the nine metabolites achieved an area under the receiver operating characteristic curve of 0.852 in the training set and 0.860 in the validation set. Conclusion ·The nine‑metabolite combined diagnostic model shows favorable diagnostic efficacy for differentiating brucellosis from rheumatoid arthritis, providing a potential auxiliary biomarker panel for clinical application.

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    Novel inflammatory and metabolic biomarkers and their association with SYNTAX score in coronary heart disease
    Ziyawudong·Bawudong , Shahemu·Anaerbai , Asiya·Aierken , Yeerkenbieke·Shadeke , LI Guangjuan, Jiang Meng
    2026, 46 (7):  946-953. 
    doi: 10.3969/j.issn.1674-8115.2026.07.013

    Abstract ( 7 )   HTML ( 0 )   PDF (1190KB) ( 2 )  

    Objective ·To investigate the correlations of the neutrophil percentage-to-albumin ratio (NPAR), monocyte-to-high-density lipoprotein ratio (MHR), fibrosis-4 index (FIB-4), and lipoprotein(a) [LP(a)] levels with the synergy between percutaneous coronary intervention with Taxus and cardiac surgery (SYNTAX) score in patients with coronary heart disease. Methods ·A total of 289 patients with coronary heart disease hospitalized in the Department of Cardiovascular Medicine at Friendship Hospital, Ili Kazakh Autonomous Prefecture, from January 2023 to January 2025 and diagnosed by coronary angiography were enrolled. According to the SYNTAX score, patients were divided into a low-risk group (n=122), an intermediate-risk group (n=86), and a high-risk group (n=81). Baseline characteristics and laboratory data were collected, including age, history of hypertension, body mass index (BMI), gender, history of diabetes, smoking history, drinking habits, triglyceride, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, LP(a), creatinine, and uric acid. Multivariate Logistic regression analysis was performed to evaluate the associations between the above factors and the SYNTAX score. Moreover, receiver operating characteristic (ROC) curve analysis was used to assess the predictive accuracy of NPAR, MHR, FIB-4, and LP(a) for the SYNTAX score. Results ·No statistically significant differences were observed among the three groups in gender, age, BMI, history of hypertension, smoking history, blood lipids, creatinine, uric acid, or other baseline data (all P>0.05). However, the prevalence of diabetes and the levels of high-density lipoprotein cholesterol, LP(a), and NPAR differed significantly among the three groups (all P<0.05). After pooling the intermediate-risk and high-risk groups into an intermediate-high-risk group (n=167), comparison with the low-risk group (n=122) revealed a significantly higher prevalence of diabetes and elevated LP(a) and NPAR levels (all P<0.05). Multivariate Logistic regression analysis revealed that diabetes (OR=1.768, 95%CI 1.024‒3.051, P=0.041), NPAR (OR=5.386, 95%CI 2.216‒13.089, P<0.001), and LP(a) (OR=1.002, 95%CI 1.000‒1.003, P=0.043) were independent factors associated with high-risk coronary artery lesions. ROC curve analysis showed that the area under the curve (AUC) of NPAR was 0.641 (95%CI 0.576‒0.705, P<0.001), and that of LP(a) was 0.572 (95%CI 0.505‒0.639, P=0.038). The combined AUC of NPAR and LP(a) was 0.665 (95%CI 0.602‒0.728, P<0.001), which demonstrated superior predictive efficacy compared with either marker alone, whereas MHR (AUC=0.530, P=0.390) and FIB-4 (AUC=0.525, P=0.478) showed no statistically significant predictive value. Conclusion ·Both NPAR and LP(a) levels were positively associated with the SYNTAX score, suggesting their potential utility as reference markers for assessing the severity of coronary artery disease.

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    Correlation between serum sugar alcohol concentrations and metabolic dysfunction-associated steatotic liver disease in the overweight/obese population
    Tan Tao, Yue Jiang, Qi Yicheng, Yang Minglan, Ma Jing
    2026, 46 (7):  954-960. 
    doi: 10.3969/j.issn.1674-8115.2026.07.014

    Abstract ( 8 )   HTML ( 0 )   PDF (988KB) ( 2 )  

    Objective ·To investigate the correlation between serum sugar alcohol levels and metabolic dysfunction-associated steatotic liver disease (MASLD) in the overweight/obese population. Methods ·This cross-sectional study enrolled 91 overweight/obese patients [body mass index (BMI)≥25 kg/m2] who visited the Obesity Clinic of Renji Hospital, Shanghai Jiao Tong University School of Medicine from July 2019 to August 2021. The participants were divided into the MASLD group (n=67) and the non-MASLD group (n=24). General data (age, sex, BMI, and blood pressure), blood biochemical parameters (liver function, renal function, blood glucose, lipid profile, insulin, etc.), and serum sugar alcohol levels (erythritol, xylitol, sorbitol, and maltitol) were collected. Serum sugar alcohol levels were quantified by gas chromatography-mass spectrometry. Magnetic resonance imaging-proton density fat fraction was used to measure liver fat content, pancreatic fat content, subcutaneous adipose tissue, and visceral adipose tissue. Spearman correlation analysis was used to analyze the correlation between serum sugar alcohol levels and liver fat content. Univariate and multivariate Logistic regression analyses were conducted to identify factors related with MASLD. Results ·A total of 91 overweight/obese patients were enrolled, with a median age of 34.00 (29.00, 42.00) years and a median BMI of 33.04 (29.60, 36.46) kg/m2. Among them, 48.35% were male. The MASLD group showed significantly higher levels of alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, uric acid, triglycerides, fasting insulin, homeostasis model assessment of insulin resistance, glycated hemoglobin, pancreatic fat content, liver fat content, visceral adipose tissue, and erythritol compared with the non-MASLD group (all P<0.05). Spearman correlation analysis showed that serum erythritol levels were positively correlated with liver fat content in the overweight/obese population (r=0.209, P=0.046), while maltitol, xylitol, and sorbitol showed no significant correlations with liver fat content. After adjustment for sex, uric acid, homeostasis model assessment of insulin resistance, and triglycerides, multivariate Logistic regression analysis revealed that elevated serum erythritol levels remained an independent risk factor for MASLD in overweight/obese patients (OR=3.02, 95%CI 1.15‒7.94, P=0.025). Conclusion ·Serum erythritol level is independently and positively associated with the risk of MASLD in the overweight/obese population.

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    Evidence-based medicine
    Immune cell traits mediate the association between alcoholic liver disease and hepatocellular carcinoma: a Mendelian randomization and mediation analysis
    Hao Fengjie, Wang Junqing, Lu Ye
    2026, 46 (7):  961-971. 
    doi: 10.3969/j.issn.1674-8115.2026.07.015

    Abstract ( 7 )   HTML ( 0 )   PDF (3007KB) ( 6 )  

    Objective ·To explore at the genetic level whether genes related to alcoholic liver disease (ALD) affect the risk of hepatocellular carcinoma (HCC) by regulating systemic immune phenotypes. Methods ·A study design combining multi-step Mendelian randomization (MR) and mediation analysis was adopted. First, cis-expression quantitative trait loci (cis-eQTL) data from the eQTLGen Consortium, ALD-related gene set from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and genome-wide association study (GWAS) summary data for HCC were integrated to identify ALD-related genes with unidirectional causal associations with HCC. Instrumental variable selection criteria included genome-wide significance thresholds, linkage disequilibrium clumping, and F-statistic calculation to exclude weak instruments. Second, MR analysis was performed on 731 peripheral blood immune cell traits to identify immune phenotypes causally associated with HCC risk. Finally, a two-step MR mediation analysis was carried out to test whether genetic effects on HCC risk were mediated through these immune phenotypes. Results ·The study identified 4 ALD-related genes [peroxisome proliferator-activated receptor γ coactivator 1-α (PPARGC1A), mitogen-activated protein kinase kinase kinase 7 (MAP3K7), aldehyde dehydrogenase 2 (ALDH2), and receptor-interacting serine/threonine protein kinase 1 (RIPK1)] with robust causal associations with HCC. Among them, ALDH2 and MAP3K7 increased HCC risk, while PPARGC1A and RIPK1 were protective. All sensitivity analyses revealed no evidence of horizontal pleiotropy or heterogeneity. Additionally, 23 peripheral blood immune cell traits were identified as being causally associated with HCC risk, involving B cells, CD8⁺ T cells, CD4⁺ T cells, regulatory T cells, and myeloid cells. Mediation analysis revealed two specific pathways: genetically predicted PPARGC1A expression partially reduced HCC risk via increased CD8dim AC cells, while the risk effect of ALDH2 on HCC was partially offset by increased naïve CD28⁺ CD45RA⁺ CD8dim %T cells. Conclusion ·Part of the ALD-related genetic risk for HCC is mediated by specific genes that alter systemic immune homeostasis. The study provides human population-based causal genetic evidence for the “ALD-immune-HCC” axis and suggests that the aforementioned genes and related immune phenotypes may serve as potential early-warning biomarkers and therapeutic targets, offering a theoretical basis for precision prevention strategies across different genetic backgrounds.

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    Identification of risk biomarkers of gallbladder cancer through plasma proteome-wide Mendelian randomization analysis
    Tang Qiuyi, Dong Wenjun, Hu Chunnan, Gong Wei
    2026, 46 (7):  972-980. 
    doi: 10.3969/j.issn.1674-8115.2026.07.016

    Abstract ( 10 )   HTML ( 0 )   PDF (1896KB) ( 2 )  

    Objective ·To systematically evaluate the potential causal associations between plasma protein levels and the risk of gallbladder cancer (GBC) through proteome-wide Mendelian randomization analysis. Methods ·GBC genome-wide association study (GWAS) summary data from Japan, Republic of Korea, Finland, and the UK were used for a multi-ethnic meta-analysis to obtain genetic effect estimates for GBC. Summary-level GWAS data of plasma proteomes from two large-scale proteomic studies (deCODE and UKBPPP) were used as exposure variables. A generalized summary-data-based Mendelian randomization (GSMR) analysis was performed to assess the causal associations between each plasma protein level and GBC risk, and significant associations were validated by two-sample Mendelian randomization (TSMR). Significant associations discovered between plasma proteins and GBC risk were also validated using colocalization analysis. Results ·GSMR analysis identified 56 plasma proteins significantly causally associated with GBC risk in the deCODE cohort (Padj<0.001), of which 31 proteins were replicated in another independent proteomic cohort (UKBPPP). TSMR further confirmed two proteins with robust causal associations with GBC. Among these two proteins, higher plasma complement receptor 2 (CR2) levels were significantly associated with a reduced risk of GBC (OR=0.258, 95%CI 0.071‒0.935, P=0.039), whereas higher plasma neural cell adhesion molecule 1 (NCAM1) levels were associated with an increased risk of GBC (OR=2.306, 95%CI 1.432‒3.713, P<0.001). The Bayesian colocalization analysis showed strong colocalization between protein quantitative trait loci (pQTL) variants of CR2 and NCAM1 and GBC risk variants. Conclusion ·Two potential risk-related biomarkers for GBC, CR2, and NCAM1, were identified in the whole plasma proteome. The elevated CR2 level is linked to a lower risk of GBC, while the elevated NCAM1 level corresponds to a higher risk.

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    Review
    Advances in alveolar bone changes and risks of alveolar surgery in patients with chronic kidney disease
    Lu Tingwei, Yuan Hao, Liu Yingli, Jiang Lingyong
    2026, 46 (7):  981-989. 
    doi: 10.3969/j.issn.1674-8115.2026.07.017

    Abstract ( 7 )   HTML ( 0 )   PDF (2703KB) ( 1 )  

    Chronic kidney disease (CKD) is a major global public health issue that significantly impacts patients′ health and quality of life. Chronic kidney disease-mineral and bone disorder (CKD-MBD), a common complication of CKD, not only leads to systemic bone metabolic abnormalities but also causes local changes such as a decrease in alveolar bone mass. Its pathological mechanisms involve multiple aspects, including calcium-phosphorus metabolism imbalance, uremic toxin damage, inflammation and oxidative stress, nutritional and metabolic disorders, as well as hormonal and growth factor dysregulation. Renal dysfunction leads to hyperphosphatemia, hypocalcemia, and secondary hyperparathyroidism, disrupting the dynamic balance between bone formation and resorption. The accumulation of uremic toxins and inflammatory cytokines induces oxidative stress, thereby suppressing osteoblast differentiation while promoting osteoclast activation. Moreover, metabolic disturbances, including vitamin D deficiency and protein-energy wasting, along with hormonal and growth factor dysregulation, impair alveolar bone homeostasis and exacerbate bone disorders. When patients with CKD undergo alveolar surgeries, such as tooth extraction and dental implant placement, they face risks of bleeding, infection, and poor wound healing. Therefore, targeted perioperative management is necessary, including multidisciplinary collaboration with nephrologists to comprehensively assess renal and coagulation functions, appropriate surgical timing, the use of minimally invasive techniques, and enhanced infection prevention. This article reviews the characteristics, mechanisms, and clinical risks of CKD-related alveolar bone changes, providing reference for related mechanism exploration and oral diagnosis and treatment.

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    Research progress in the impact of elevated follicle-stimulating hormone levels on metabolic changes in postmenopausal women
    Lin Xuefeng, Lai Dongmei
    2026, 46 (7):  990-996. 
    doi: 10.3969/j.issn.1674-8115.2026.07.018

    Abstract ( 15 )   HTML ( 0 )   PDF (1418KB) ( 9 )  

    The hypothalamic-pituitary-ovarian axis (HPO) is the core endocrine system regulating the female menstrual cycle. The components of the HPO operate within a complex hormonal regulatory network to maintain reproductive health. Follicle-stimulating hormone (FSH), secreted by the pituitary gland, is essential for follicular development and steroidogenesis, whereas estrogen provides negative feedback to the hypothalamus and pituitary to preserve the dynamic balance of the HPO. With the onset of menopause, ovarian failure leads to a marked decline in estrogen levels, weakening this feedback loop and resulting in sustained elevations of circulating FSH. Accumulating evidence indicates that high FSH levels in postmenopausal women have effects extending beyond the reproductive system, acting through the follicle-stimulating hormone receptor (FSHR) expressed in adipose tissue, pancreatic islets, and bone to participate in energy metabolic remodeling and the regulation of metabolic homeostasis. Current findings suggest that elevated FSH is associated with lipid metabolic disturbances, adipose tissue inflammation, and an increased risk of cardiovascular disease. In glucose metabolism, FSH acts through FSHR expressed on pancreatic β cells to inhibit glucose-stimulated insulin secretion (GSIS), ultimately leading to impaired glucose homeostasis. In bone metabolism, FSH directly enhances osteoclast activity and promotes bone resorption, with part of this effect being independent of estrogen deficiency. This review summarizes advances in understanding the roles of elevated FSH levels in lipid, glucose, and bone metabolism in postmenopausal women and highlights the potential clinical implications and remaining challenges in this field.

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