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.

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.

Objective To investigate the effect of miR-206 on LPS-induced apoptosis and inflamma-tory response in H9c2 cardiomyocytes.Methods An inflammatory injury model was established in H9c2 cells with treatment of 10 μg/ml LPS.Then the cells were divided into control group,LPS group,LPS1 group(LPS+anti-negative control),LPS2 group(LPS+anti-miR-206),LPS3 group(LPS+miR-206 negative control),and LPS4 group(LPS+miR-206 mimic).The LPS1,LPS2,LPS3,and LPS4 groups(n=3)were transfected with anti-negative control,anti-miR-206 se-quence,miR-negative control,or miR-206 mimic sequence,using Lipofectamine 3000 reagent.CCK-8 and EdU assays were used to detect cell proliferation,Caspase-Glo 3/7 activity kit was em-ployed to measure Caspase-3/7 activity,and ELISA was utilized to test the secretion of inflamma-tory cytokines,TNF-α,IL-1β,and IL-6 in the cells.Results Compared with the control group,the LPS group had significantly reduced cell survival rate and EDU-positive cell rate,while elevated miR-206 expression,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65(P＜0.05,P＜0.01).The expression of miR-206,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65 were obviously reduced,while the cell survival rate and EDU-positive cell rate were notably increased in the LPS 2 group than the LPS group(P＜0.05).In the LPS 4 group,the activity of Caspase-3/7 was remarkably stronger than the LPS group(1586±58 vs 816±51,P＜0.05).Conclusion Suppression of miR-206 can effectively inhibit LPS-induced apoptosis and inflammatory response in H9c2 cardiomyocytes.

As new evidence emerges, treatment strategies toward the functional cure of chronic hepatitis B are evolving. In 2019, a panel of national hepatologists published a Consensus Statement on the functional cure of chronic hepatitis B. Currently, an international group of hepatologists has been assembled to evaluate research since the publication of the original consensus, and to collaboratively develop the updated statements. The 2.0 Consensus was aimed to update the original consensus with the latest available studies, and provide a comprehensive overview of the current relevant scientific literatures regarding functional cure of hepatitis B, with a particular focus on issues that are not yet fully clarified. These cover the definition of functional cure of hepatitis B, its mechanisms and barriers, the effective strategies and treatment roadmap to achieve this endpoint, in particular new surrogate biomarkers used to measure efficacy or to predict response, and the appropriate approach to pursuing a functional cure in special populations, the development of emerging antivirals and immunomodulators with potential for curing hepatitis B. The statements are primarily intended to offer international guidance for clinicians in their practice to enhance the functional cure rate of chronic hepatitis B.

To analyze the prevalence, drug resistance and whole genome characteristics of Mycoplasma pneumoniae (MP) in respiratory throat swab samples of hospitalized children with pneumonia in Suzhou City from 2023 to 2024. Throat swab samples of hospitalized children aged 0-14 years old with pneumonia in Suzhou were collected from September 2023 to September 2024. Real-time fluorenscence quantitative PCR technology was used to detect MP nucleic acid. The results showed that the positive rate of MP in 3 235 samples was 22.44% (726/3 235), with a rate of 55.00% in week 47 of 2023. The positive rate of MP increased with age ( χ2=45.842, P<0.001). The study selected MP nucleic acid test positive samples from week 20 (5.13-5.19) to week 23 (6.3-6.9) of 2024 for isolation, culture and resistance phenotype detection. About 31 MP strains were successfully isolated and cultured, all of which were resistant to macrolides. The next-generation sequencing technology and nanopore sequencing technology were used for genome sequencing. All 31 strains carried the A2063G mutation, with the main prevalent genotype being the P1-1, and the main mlST type being the ST3. Despite the overall genomic similarity between strains being over 99%, there were significant differences between the P1-1 and P1-2 strains in the P1 gene region. In summary, from 2023 to 2024, the main MP type prevalent in Suzhou City is the P1-1 genotype. All isolated MP strains carry an A2063G resistance site mutation and are resistant to macrolides, requiring continuous monitoring and further research.

As new evidence emerges, treatment strategies toward the functional cure of chronic hepatitis B are evolving. In 2019, a panel of national hepatologists published a Consensus Statement on the functional cure of chronic hepatitis B. Currently, an international group of hepatologists has been assembled to evaluate research since the publication of the original consensus, and to collaboratively develop the updated statements. The 2.0 Consensus was aimed to update the original consensus with the latest available studies, and provide a comprehensive overview of the current relevant scientific literatures regarding functional cure of hepatitis B, with a particular focus on issues that are not yet fully clarified. These cover the definition of functional cure of hepatitis B, its mechanisms and barriers, the effective strategies and treatment roadmap to achieve this endpoint, in particular new surrogate biomarkers used to measure efficacy or to predict response, and the appropriate approach to pursuing a functional cure in special populations, the development of emerging antivirals and immunomodulators with potential for curing hepatitis B. The statements are primarily intended to offer international guidance for clinicians in their practice to enhance the functional cure rate of chronic hepatitis B.

OBJECTIVES: To investigate the effect of Haematococcus pluvialis (HP) on bleomycin (BLM)-induced pulmonary fibrosis in mice and on TGF-β1-induced human fetal lung fibroblasts (HFL1). METHODS: Thirty male C57BL/6 mice were randomly divided into control group, BLM-induced pulmonary fibrosis model group, low- and high-dose HP treatment groups (3 and 21 mg/kg, respectively), and 300 mg/kg pirfenidone (positive control) group. The effects of drug treatment for 21 days were assessed by examining respiratory function, lung histopathology, and expression of fibrosis markers in the lung tissues of the mouse models. In TGF-β1-induced HFL1 cell cultures, the effects of treatment with 120, 180 and 240 μg/mL HP or 1.85 μg/mL pirfenidone for 48 h on expression levels of fibrosis markers were evaluated. Transcriptome analysis was carried out using the control cells and cells treated with TGF-β1 and 240 μg/mL HP. RESULTS: HP obviously alleviated BLM-induced lung function damage and fibrotic changes in mice, evidenced by improved respiratory function, lung tissue morphology and structure, inflammatory infiltration, and collagen deposition and reduced expressions of fibrotic proteins. HP at the high dose produced similar effect to PFD. In TGF-β1-induced HFL1 cells, treatment with 240 μg/mL HP significantly reduced the mRNA and protein expression levels of α-SMA and FN. Transcriptome analysis revealed that multiple key genes and pathways mediated the protective effect of HP against pulmonary fibrosis. CONCLUSIONS HP alleviates pulmonary fibrosis in both the mouse model and cell model, possibly as the result of the synergistic effects of its multiple active components.
Animals ; Pulmonary Fibrosis/chemically induced* ; Bleomycin/adverse effects* ; Mice, Inbred C57BL ; Male ; Mice ; Fibroblasts/drug effects* ; Lung/pathology* ; Transforming Growth Factor beta1/pharmacology* ; Myofibroblasts/drug effects* ; Humans ; Pyridones

Objective To investigate the effect of Haematococcus pluvialis(HP)on bleomycin(BLM)-induced pulmonary fibrosis in mice and on TGF-β1-induced human fetal lung fibroblasts(HFL1).Methods Thirty male C57BL/6 mice were randomly divided into control group,BLM-induced pulmonary fibrosis model group,low-and high-dose HP treatment groups(3 and 21 mg/kg,respectively),and 300 mg/kg pirfenidone(positive control)group.The effects of drug treatment for 21 days were assessed by examining respiratory function,lung histopathology,and expression of fibrosis markers in the lung tissues of the mouse models.In TGF-β1-induced HFL1 cell cultures,the effects of treatment with 120,180 and 240 μg/mL HP or 1.85 μg/mL pirfenidone for 48 h on expression levels of fibrosis markers were evaluated.Transcriptome analysis was carried out using the control cells and cells treated with TGF-β1 and 240 μg/mL HP.Results HP obviously alleviated BLM-induced lung function damage and fibrotic changes in mice,evidenced by improved respiratory function,lung tissue morphology and structure,inflammatory infiltration,and collagen deposition and reduced expressions of fibrotic proteins.HP at the high dose produced similar effect to PFD.In TGF-β1-induced HFL1 cells,treatment with 240 μg/mL HP significantly reduced the mRNA and protein expression levels of α-SMA and FN.Transcriptome analysis revealed that multiple key genes and pathways mediated the protective effect of HP against pulmonary fibrosis.Conclusion HP alleviates pulmonary fibrosis in both the mouse model and cell model,possibly as the result of the synergistic effects of its multiple active components.

Objective:To analyze the dual gain effect of hyperbaric oxygen combined with vestibular rehabilitation training on patients with tinnitus and dizziness.Methods:This study was a retrospective analysis,and 90 patients with tinnitus and dizziness who visited the outpatient department of our hospital from January 2022 to January 2025 were selected to carry out the study,and were divided into observation group and control group according to the treatment methods,with 45 cases in each group.The matched group patients were treated with conventional medication,while the observation group received hyperbaric oxygen combined with vestibular rehabilitation training on the basis of the matched group.The clinical efficacy of the two groups was compared,including the concentration of oxygenated hemoglobin in the cerebral cortex,vestibular function,severity of tinnitus,and severity of dizziness before and after treatment.Results:The total effective rate of the observation group was higher than the matched group(P＜0.05);Post-treatment,there was no significant change in the concentration of oxygenated hemoglobin in the parietal and occipital cortex between the two groups(P＞0.05).However,the concentration of oxygenated hemoglobin in the temporal lobe,dorsolateral prefrontal cortex,and frontal polar cortex increased,and the observation group was higher than the matched group(P＜0.05);Post-treatment,the values of hemiparesis,total area of center of gravity movement(CA),and total length of center of gravity movement(PL)in both groups decreased,and the observation group was lower than the matched group(P＜0.05);Post-treatment,the severity score of tinnitus in both groups decreased,and the observation group was lower than that in the matched group(P＜0.05);Post-treatment,the degree score of vertigo in both groups decreased,and the observation group was lower than that in the matched group(P＜0.05).Conclusion:Hyperbaric oxygen combined with vestibular rehabilitation training has a significant dual gain effect on patients with tinnitus and dizziness,which can improve clinical efficacy,promote cortical neural activity,improve vestibular function,and thereby alleviate the severity of tinnitus and dizziness.

As new evidence emerges, treatment strategies toward the functional cure of chronic hepatitis B are evolving. In 2019, a panel of national hepatologists published a Consensus Statement on the functional cure of chronic hepatitis B. Currently, an international group of hepatologists has been assembled to evaluate research since the publication of the original consensus, and to collaboratively develop the updated statements. The 2.0 Consensus was aimed to update the original consensus with the latest available studies, and provide a comprehensive overview of the current relevant scientific literatures regarding functional cure of hepatitis B, with a particular focus on issues that are not yet fully clarified. These cover the definition of functional cure of hepatitis B, its mechanisms and barriers, the effective strategies and treatment roadmap to achieve this endpoint, in particular new surrogate biomarkers used to measure efficacy or to predict response, and the appropriate approach to pursuing a functional cure in special populations, the development of emerging antivirals and immunomodulators with potential for curing hepatitis B. The statements are primarily intended to offer international guidance for clinicians in their practice to enhance the functional cure rate of chronic hepatitis B.