AIM: To investigate the correlation of serum leukocyte-derived chemotaxin 2(LECT2)and complement C1q/tumor necrosis factor-related protein 5(CTRP5)with diabetic retinopathy(DR).METHODS:A single-center cross-sectional analysis was conducted on 138 patients with type 2 diabetes mellitus(T2DM)admitted to our hospital from April 2023 to April 2025. According to the diagnostic results, they were divided into a non-DR group(60 cases)and a DR group(78 cases). The DR group was further divided into a proliferative DR(PDR)group(29 cases)and a non-proliferative DR(NPDR)group(49 cases). Pearson correlation analysis was used to assess the correlation of LECT2 and CTRP5 with glucose-lipid metabolism indicators. Logistic regression analysis was conducted to identify the influencing factors for the occurrence of DR in T2DM patients. ROC curve analysis was performed to evaluate the diagnostic value of serum LECT2 and CTRP5 for DR in T2DM patients.RESULTS: A comparison of general patient data between the two groups showed no significant differences. The DR group had higher serum levels of LECT2, CTRP5, total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), and homeostatic model assessment of insulin resistance(HOMA-IR)than the non-DR group, while high-density lipoprotein cholesterol(HDL-C)was lower(all P<0.05). The serum levels of LECT2 and CTRP5 in the PDR group were higher than those in the NPDR group(all P<0.05). Serum LECT2 and CTRP5 positively correlated with TC, TG, LDL-C, and HOMA-IR, and negatively correlated with HDL-C(all P<0.001). Logistic results showed that duration of diabetes, TC, TG, LDL-C, HOMA-IR, LECT2, and CTRP5 were risk factors for DR occurrence in T2DM patients(all P<0.05). The ROC curve showed that the AUCs for serum LECT2, CTRP5 alone, and combined in diagnosing DR in T2DM patients were 0.830, 0.839, and 0.915, respectively. The AUC for the combined diagnosis was higher than that of LECT2 or CTRP5 alone according to the DeLong test(Z=2.818, 2.824, P=0.015, 0.012).CONCLUSION:Serum LECT2 and CTRP5 levels are closely related to the development of DR in T2DM patients, and joint detection has certain clinical value in diagnosing DR in T2DM patients.

AIM:To investigate the predictive value of aqueous humor stromal cell-derived factor-1(SDF-1)and epithelial neutrophil-activating peptide-78(ENA78)for visual impairment following vitrectomy in patients with diabetic retinopathy(DR).METHODS: Patients with DR who underwent vitrectomy in the ophthalmology department of our hospital from March 2022 to March 2024 were enrolled as study group. According to the results of uncorrected visual acuity examination during the 1-year follow-up after surgery, they were further divided into visual impairment group and good visual acuity group. In addition, patients with cataracts who were treated in our hospital during the same period were recruited as control group. The levels of SDF-1 and ENA78 in aqueous humor were detected by enzyme-linked immunosorbent assay(ELISA). Pearson correlation analysis was used to evaluate the correlation of indicators. Multivariate Logistic regression analysis was performed to identify the influencing factors of postoperative visual impairment in DR patients. Relative risk analysis was conducted to assess the impact of different aqueous humor SDF-1 and ENA78 levels on postoperative visual impairment in DR patients. Receiver operating characteristic(ROC)curve analysis was applied to determine the predictive value of aqueous humor SDF-1 and ENA78 levels for postoperative visual impairment in DR patients.RESULTS: Totally 156 cases(156 eyes)were enrolled in the study group, including 87 males and 69 females, with a mean age of 60.65±8.91 years old; and 91 cases(91 eyes)were enrolled in the control group, including 53 males and 38 females, with a mean age of 58.72±8.43 years old. The levels of SDF-1 and ENA78 in aqueous humor of the study group were significantly higher than those of the control group(P<0.001). According to the results of uncorrected visual acuity during the 1-year follow-up after surgery, the study group was further divided into the visual impairment group(n=49)and the good visual acuity group(n=107), including 28 males and 21 females in the visual impairment group, with a mean age of 63.73±7.41 years, and 59 males and 48 females in the good visual acuity group, with a mean age of 59.24±8.47 years. The age and glycated hemoglobin(HbA1c)levels of the visual impairment group were higher than those of the good visual acuity group(all P<0.05). Compared with the good visual acuity group, the levels of SDF-1 and ENA78 in aqueous humor of the visual impairment group were significantly elevated(all P<0.05). A positive correlation was found in the study group between the levels of SDF-1, ENA78 in aqueous humor and HbA1c(r=0.314, 0.362, all P<0.05). Elevated levels of aqueous humor SDF-1(OR=2.936, 95% CI: 1.628-5.296)and ENA78(OR=3.092, 95% CI: 1.738-5.502)were identified as risk factors for postoperative visual impairment in DR patients(all P<0.05). High levels of SDF-1 and ENA78 increased the risk of postoperative visual impairment by 1.958-fold and 2.030-fold compared with the low-level groups, respectively; the area under the curve(AUC)of combined detection of aqueous humor SDF-1 and ENA78 for predicting postoperative visual impairment in DR patients was 0.936, which was superior to the predictive value of SDF-1 or ENA78 alone(Zcombination-SDF-1=2.850, Zcombination-ENA78=2.445, all P<0.05).CONCLUSION: Elevated levels of aqueous humor SDF-1 and ENA78 are risk factors for postoperative visual impairment in DR patients, and the combined detection of SDF-1 and ENA78 has high clinical application value for predicting postoperative visual impairment in DR patients.

Hepatocellular carcinoma (HCC) develops within a profoundly immunosuppressive tumor immune microenvironment (TIME), which limits the efficacy of immunotherapy. Polarization of tumor-associated macrophages (TAMs) toward a pro-tumorigenic M2 phenotype is a major driver of immune escape. Ferroptosis, an iron-dependent regulated cell death program, intersects with hepatic iron metabolism and immune regulation and thus offers promising points of therapeutic intervention. This review systematically elucidates the mechanistic role of TAM ferroptosis in HCC immune evasion and highlights a “bidirectional regulation” intervention strategy grounded in the Traditional Chinese medicine (TCM) principle of “fortifying healthy qi and eliminating pathogens” (Fuzheng Quxie). This strategy employs “eliminating pathogens” (Quxie) approaches to exploit the metabolic vulnerability of M2-like TAMs and precisely induce their ferroptosis. Moreover, it utilizes “fortifying healthy qi” (Fuzheng) approaches to protect M1-like TAMs and CD8⁺ T cells from oxidative damage. This parallel “induction-protection” paradigm demonstrates the unique advantages of TCM in systemically remodeling TIME through multitarget synergistic actions. Accordingly, precision regulation of TAM ferroptosis based on the Fuzheng Quxie theory represents a promising integrative Chinese-Western medicine strategy for overcoming current bottlenecks in HCC immunotherapy, although its clinical translational potential warrants further validation.

Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

This article reports a case of a male patient with anti-N-methyl-D-aspartate receptor （NMDAR） encephalitis comorbid with early-onset Alzheimer's disease （AD）. The patient presented with depression， hallucinations and delusions syndrome， and cognitive impairment， and was admitted to a psychiatric hospital. Through multidisciplinary consultation， the diagnosis was confirmed based on positive biological and pathological biomarkers. The patient's symptoms improved after receiving a therapeutic regimen comprising high-dose glucocorticoid therapy and psychiatric-related treatments. Based on relevant current research findings domestically and internationally， this article reviews the pathogenesis and clinical manifestations of early-onset AD and anti-NMDAR encephalitis， in order to enhance the discrimination ability of psychiatric specialists for such patients and improve the diagnosis and treatment level. ［Funded by Medical Research Project of Chengdu City （number， 2024403）］

Objective To analyze the genomic characteristics of Legionella pneumophila (L. pneumophila) isolated from industrial circulating cooling water in workplace. Methods A total of 20 L. pneumophila strains isolated from industrial circulating cooling water collected from eight chemical enterprises were selected as the research subjects. Whole-genome sequencing was performed on the strains, followed by multilocus sequence typing (MLST), whole-genome single nucleotide polymorphism analysis, virulence factor profiling, and antibiotic resistance gene detection. Results The dominant MLST typing among the 20 L. pneumophila strains was ST2. Serogroup 1 (Lp1) strains exhibited high genetic diversity. Strains from different enterprises showed substantial genomic diversity, suggesting possible inter-enterprise dissemination. Each isolate harbored 175-295 virulence genes; among them, virulence genes were identified, and 139 genes were detected in all the 20 strains. These genes primarily included those related to the Dot/Icm type Ⅳ bacterial secretion system, flagellar synthesis and motility, and type Ⅳ pili. The number of virulence genes carried by L. pneumophila strains of different ST types differed (P<0.01), with unidentified sequence types harboring more virulence genes than other STs (P<0.01). A total of four resistance genes were predicted, including aminoglycoside phosphotransferase-encoding gene APH(9)-Ia, carbapenemase-encoding gene OXA-29, and efflux pump genes-encoding genes LpeA and LpeB, with detection rates of 100.00%, 100.00%, 10.00%, and 10.00%, respectively. These genes may confer resistance to cephalosporins, penicillins, and aminoglycosides, and all strains were multidrug resistance. Conclusion L. pneumophila isolated from industrial recirculating cooling water exhibits high genetic diversity, strong pathogenicity, and significant multidrug resistance potential.

Objective To systematically evaluate the similarities between six-gene-modified pig red blood cells and human red blood cells in multiple dimensions, including cell morphology, hematological parameters, microbial safety and serological compatibility, and to verify the feasibility of using six-gene-modified pig red blood cells as an alternative for blood transfusion in cases of traumatic hemorrhage. Methods Blood samples from O-type human donor, six-gene-modified O-type pig and wild-type O-type pig were collected. The pig gene-modified phenotype was identified by flow cytometry. The morphology was observed using field emission scanning electron microscopy and the diameter was measured. Hematological tests, including blood routine, osmotic fragility test, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) content and partial pressure of oxygen at 50% hemoglobin saturation (P50), were conducted to assess the deformability of red blood cells. Real-time fluorescent quantitative polymerase chain reaction was used to screen for zoonotic viruses, and cross-matching was performed using column gel method, condensation amine method and saline tube method. Results The six-gene-modified pig red blood cells successfully knocked out the α-Gal, Neu5Gc and Sda antigens and expressed hCD55. Wild-type pig red blood cells expressed these three antigens but did not express human complement regulatory proteins. All three types of red blood cells presented a biconcave discoid morphology. The diameter of O-type human red blood cells was larger than that of the six-gene-modified porcine red blood cells and wild-type pig red blood cells (all P < 0.05). The average red blood cell volume and average red blood cell hemoglobin of O-type humans were higher than those of the six-gene-modified pig red blood cells, and the distribution width-variable coefficient of O-type human red blood cells was lower than that of the six-gene-modified pig red blood cells (all P < 0.05). There were no statistically significant differences in the permeability fragility, ATP content, 2,3-DPG content, P50 and red blood cell deformability indicators of the three types of red blood cells (all P > 0.05). The screening for zoonotic viruses in human and animal cells of the six-gene-modified pigs and wild-type pigs was negative. The cross-matching results showed that in the column gel method and condensation amine method, the primary and secondary sides of the six-gene-modified pig red blood cells did not agglutinate with human red blood cells, while the wild-type pig red blood cells agglutinated with human red blood cells in all cross-matching methods. Conclusions The six-gene-modified pig red blood cells are highly similar to human red blood cells in terms of morphology, key hematological parameters, energy metabolism, oxygen-carrying capacity, mechanical properties, microbial safety and serological compatibility, providing an experimental basis for the clinical transformation of xenotransfusion.

The present study aimed to explore whether hydrogen sulfide (H₂S) improved hypoxic pulmonary hypertension (HPH) in rats by inhibiting aerobic glycolysis-pyroptosis. Male Sprague-Dawley (SD) rats were randomly divided into normal group, normal+NaHS group, hypoxia group, and hypoxia+NaHS group, with 6 rats in each group. The control group rats were placed in a normoxic (21% O₂) environment and received daily intraperitoneal injections of an equal volume of normal saline. The normal+NaHS group rats were placed in a normoxic environment and intraperitoneally injected with 14 μmol/kg NaHS daily. The hypoxia group rats were placed in a hypoxia chamber, and the oxygen controller inside the chamber maintained the oxygen concentration at 9% to 10% by controlling the N₂ flow rate. An equal volume of normal saline was injected intraperitoneally every day. The hypoxia+NaHS group rats were also placed in an hypoxia chamber and intraperitoneally injected with 14 μmol/kg NaHS daily. After the completion of the four-week modeling, the mean pulmonary artery pressure (mPAP) of each group was measured using right heart catheterization technique, and the right ventricular hypertrophy index (RVHI) was weighed and calculated. HE staining was used to observe pathological changes in lung tissue, Masson staining was used to observe fibrosis of lung tissue, and Western blot was used to detect protein expression levels of hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate kinase isozyme type M2 (PKM2), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), GSDMD-N-terminal domain (GSDMD-N), Caspase-1, interleukin-1β (IL-1β) and IL-18 in lung tissue. ELISA was used to detect contents of IL-1β and IL-18 in lung tissue. The results showed that, compared with the normal control group, there were no significant changes in all indexes in the normal+NaHS group, while the hypoxia group exhibited significantly increased mPAP and RVHI, thickened pulmonary vascular wall, narrowed lumen, increased collagen fibers, up-regulated expression levels of aerobic glycolysis-related proteins (HK2 and PKM2), up-regulated expression levels of pyroptosis-related proteins (NLRP3, GSDMD-N, Caspase-1, IL-1β, and IL-18), and increased contents of IL-1β and IL-18. These changes of the above indexes in the hypoxia group were significantly reversed by NaHS. These results suggest that H₂S can improve rat HPH by inhibiting aerobic glycolysis-pyroptosis.
Animals ; Rats, Sprague-Dawley ; Male ; Hypertension, Pulmonary/metabolism* ; Glycolysis/drug effects* ; Hydrogen Sulfide/therapeutic use* ; Hypoxia/complications* ; Rats ; Pyroptosis/drug effects*

The circadian clock plays a critical role in regulating various physiological processes, including gene expression, metabolic regulation, immune response, and the sleep-wake cycle in living organisms. Post-translational modifications (PTMs) are crucial regulatory mechanisms to maintain the precise oscillation of the circadian clock. By modulating the stability, activity, cell localization and protein-protein interactions of core clock proteins, PTMs enable these proteins to respond dynamically to environmental and intracellular changes, thereby sustaining the periodic oscillations of the circadian clock. Different types of PTMs exert their effects through distincting molecular mechanisms, collectively ensuring the proper function of the circadian system. This review systematically summarized several major types of PTMs, including phosphorylation, acetylation, ubiquitination, SUMOylation and oxidative modification, and overviewed their roles in regulating the core clock proteins and the associated pathways, with the goals of providing a theoretical foundation for the deeper understanding of clock mechanisms and the treatment of diseases associated with circadian disruption.
Protein Processing, Post-Translational/physiology* ; Circadian Rhythm/physiology* ; Humans ; Animals ; CLOCK Proteins/physiology* ; Circadian Clocks/physiology* ; Phosphorylation ; Acetylation ; Ubiquitination ; Sumoylation

The aim of this study was to investigate the contribution of lung zinc ions to pathogenesis of lung ischemia/reperfusion (I/R) injury in rats. Male Sprague Dawley (SD) rats were randomly divided into control group, lung I/R group (I/R group), lung I/R + low-dose zinc chloride group (LZnCl₂+I/R group), lung I/R + high-dose ZnCl₂ group (HZnCl₂+I/R group), lung I/R + medium-dose ZnCl₂ group (MZnCl₂+I/R group) and TPEN+MZnCl₂+I/R group (n = 8 in each group). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the concentration of zinc ions in lung tissue. The degree of lung tissue injury was analyzed by observing HE staining, alveolar damage index, lung wet/dry weight ratio and lung tissue gross changes. TUNEL staining was used to detect cellular apoptosis in lung tissue. Western blot and RT-qPCR were used to determine the protein expression levels of caspase-3 and ZIP8, as well as the mRNA expression levels of zinc transporters (ZIP, ZNT) in lung tissue. The mitochondrial membrane potential (MMP) of lung tissue was detected by JC-1 MMP detection kit. The results showed that, compared with the control group, the lung tissue damage, lung wet/dry weight ratio and alveolar damage index were significantly increased in the I/R group. And in the lung tissue, the concentration of Zn²⁺ was markedly decreased, while the cleaved caspase-3/caspase-3 ratio and apoptotic levels were significantly increased. The expression levels of ZIP8 mRNA and protein were down-regulated significantly, while the mRNA expression of other zinc transporters remained unchanged. There was also a significant decrease in MMP. Compared with the I/R group, both MZnCl₂+I/R group and HZnCl₂+I/R group exhibited significantly reduced lung tissue injury, lung wet/dry weight ratio and alveolar damage index, increased Zn²⁺ concentration, decreased ratio of cleaved caspase-3/caspase-3 and apoptosis, and up-regulated expression levels of ZIP8 mRNA and protein. In addition, the MMP was significantly increased in the lung tissue. Zn²⁺ chelating agent TPEN reversed the above-mentioned protective effects of medium-dose ZnCl₂ on the lung tissue in the I/R group. The aforementioned results suggest that exogenous administration of ZnCl₂ can improve lung I/R injury in rats.
Animals ; Reperfusion Injury/pathology* ; Male ; Rats, Sprague-Dawley ; Rats ; Chlorides/administration & dosage* ; Lung/pathology* ; Zinc Compounds/administration & dosage* ; Apoptosis/drug effects* ; Caspase 3/metabolism* ; Cation Transport Proteins/metabolism*