ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

ObjectiveTo establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus (MHV), mouse parvovirus (MPV), and reovirus type 3 (Reo-3) in laboratory mice. MethodsRecombinant plasmid standards were constructed using conserved genomic regions of the three target viruses. Specific primers and probes were designed, and a triplex TaqMan quantitative PCR (qPCR) system was optimized through preliminary experiments. Sensitivity was evaluated using 10-fold serial dilutions (10¹-10⁷ copies/μL). Intra- and inter-assay repeatability were assessed via reproducibility experiments. ResultsA strong linear correlation (R ²>0.99) was observed between copy number and Ct value at template concentrations of 10²-10⁷ copies/μL. Intra-assay and inter-assay coefficients of variation (CV) were both below 5%. No cross-reactivity was observed with mouse cytomegalovirus (MCMV), vesicular stomatitis virus (VSV), Sendai virus (SeV), or pneumonia virus of mice (PVM). ConclusionThe triplex TaqMan quantitative PCR method established in this study has high sensitivity, good repeatability, and strong specificity. It enables the rapid, specific, and sensitive detection of MHV, MPV, and Reo-3, allows the simultaneous detection of multiple pathogens in a single tube, and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations, thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.

Objective To evaluate the safety and efficacy of profunda femoral vein thrombosis clearance in endovascular treatment of acute lower extremity deep venous thrombosis (DVT). Methods From January 2022 to September 2023, clinical data of acute lower extremity DVT patients who underwent percutaneous mechanical thrombectomy (PMT) for profunda femoral vein thrombosis were retrospectively analyzed. The 12-month primary patency rate of iliofemoral vein and incidence of post-thrombotic syndrome(PTS) were analyzed. Results Twenty-two patients were included in the study, and all patients underwent PMT for profunda femoral vein thrombosis. Five patients were treated through the contralateral common femoral vein via crossover approach, and 17 patients were treated through the contralateral common femoral vein combined with the ipsilateral deep vein approach. The 12-month primary patency rate of iliofemoral vein was 90.9% (20/22), and the incidence of PTS was 9.1% (2/22). Conclusions For patients with acute lower extremity DVT undergoing endovascular treatment, PMT for profunda femoral vein thrombosis is safe and effective, achieving good primary patency of iliofemoral vein and low incidence of PTS.

Objective To investigate the clinical efficacy of Shuyao Huoxue Tongluo Decoction combined with meridian acupoint massage in treating lumbar disc herniation(LDH)patients with qi-stagnation and blood-stasis syndrome,and to analyze its effects on serological indicators.Methods A total of 120 LDH patients with qi-stagnation and blood-stasis syndrome admitted to Huizhou Hospital of Guangzhou University of Chinese Medicine from December 2023 to December 2024 were enrolled.The patients were randomly divided into control group and study group using a random number table,with 60 cases in each group.The control group received conventional treatment(including lumbar traction and drug therapy)combined with meridian acupoint massage,while the study group received additional Shuyao Huoxue Tongluo Decoction based on the treatment for the control group.Before and after treatment,the scores of traditional Chinese Medicine(TCM)syndrome,Visual Analogue Scale(VAS)scores for pain,Japanese Orthopedic Association(JOA)scores for lumbar function,Oswestry Disability Index(ODI)scores,and serum levels of tumor necrosis factor α(TNF-α)and interleukin 6(IL-6)in the two groups were compared.After treatment,the clinical efficacy was evaluated between groups.Results(1)After 20 days of treatment,the total effective rate was 95.00%(57/60)in the study group versus 81.67%(49/60)in the control group(tested by chi-square test,P＜0.05),demonstrating significantly superior therapeutic effects in the study group.(2)After treatment,both groups showed significant reduction in TCM syndrome scores(P＜0.01),and the study group exhibited markedly decrease compared to the control group(P＜0.01).(3)After treatment,the VAS scores for pain in both groups were significantly decreased compared to those before treatment(P＜0.01),and the study group exhibited obvious decrease of the scores than the control group(P＜0.01).(4)After treatment,JOA scores for lumbar function in both groups were increased(P＜0.05),while Oswestry Disability Index(ODI)scores were decreased(P＜0.05),compared to those before treatment,and the study group demonstrated more significant improvements in both JOA score elevation and ODI score reduction(P＜0.05).(5)For serum inflammatory factors,interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels in both groups were significantly reduced(P＜0.05)after treatment,and the study group showed more pronounced decrease in both markers(P＜0.05).Conclusion Compared with meridian acupoint massage alone,the combined therapy of Shuyao Huoxue Tongluo Decoction and meridian acupoint massage is effective on significantly alleviating clinical symptoms,reducing serum inflammatory factors,and enhancing overall therapeutic outcomes in LDH patients with qi-stagnation and blood-stasis syndrome.

Objective To investigate the biological functions of the ERG3 gene in Candida albicans and its potential value in antifungal therapy. Methods The ERG3 null mutant was constructed by the CRISPR/Cas9 technology. Gas chromatography-mass spectrometry, microbroth dilution method, hyphal induction and mouse systemic infection models were carried out to evaluate sterol metabolism, drug susceptibility, hyphal formation ability and pathogenicity in C. albicans. Results The disruption of the ERG3 gene led to disordered sterol metabolism in C. albicans with a significant increased level of episterol, 14α-methylfecosterol and ergosta-7,22-dienol. The ERG3 null mutant exhibited significantly reduced susceptibility to antifungal azole and polyene drugs, which suggested that ERG3 involve in regulating drug resistance. Although the disruption of ERG3 inhibited hyphal growth and biofilm formation, it did not significantly alter the pathogenicity of the strain in a mouse model of systemic fungal infection. Conclusion The ERG3 gene was a key regulator in the ergosterol synthesis pathway in C. albicans. Its deletion induced multi-drug resistance by reshaping sterol metabolism, while pathogenicity maintenance depended on compensatory mechanisms. This study provided critical insights for developing antifungal drugs targeting sterol metabolism and overcoming drug resistance.

Objective To investigate the feasibility of using the cell-free DNA(cfDNA)carrying tissue-specific mutations as biomarkers for assessing the severity of exercise-induced tissue injury.Methods Based on a public gene expression database,ten tissue-specific and highly-expressed genes were selected for each of ten different human tissues.A total of 34 young healthy volunteers were recruited,and their peripheral blood samples were collected after running 5 km per day for one week.Genomic DNA from leukocytes,plasma exosomal RNA,and plasma cfDNA were extracted and subjected to high-throughput sequencing respectively.Tissue-specific somatic mutations in plasma exosomal RNA were identified,and the relationship between cfDNA carrying these mutations and traditional protein biomarkers was analyzed.Results Tissue-specific cfDNA mutations related to five tissues(myocardium,skeletal muscle,intestine,stomach,and kidney)were detected in more than five volunteers.The correlation coefficients between total plasma cfDNA levels and the levels of tissue-specific protein biomarkers associated with these five tissues were less than 0.3(n=34,r=-0.51-0.28,P=0.0022-0.65).Notably,for each specific tissue,the levels of tissue-specific cfDNA mutations were positively correlated with the corresponding protein biomarker levels,and correlation coefficients were over 0.8(n=7-13,r=0.81-0.92,P=9.0×10-4-0.020).Conclusion Tissue-specific cfDNA mutations may serve as novel potential biomarkers for non-invasive evaluation of exercise-related tissue injury.

Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.

Objective:To compare the diagnostic efficacy of 18F-fibroblast activation protein inhibitor (FAPI)-42 PET/CT and 18F-FDG PET/CT for bone metastasis in patients with malignant tumors. Methods:From January 2022 to October 2023, the data of 238 patients (160 males, 78 females; age: 58(50, 66) years) with various malignant tumors who underwent both 18F-FAPI-42 and 18F-FDG PET/CT imaging at Nanfang Hospital, Southern Medical University were retrospectively reviewed. An abnormal focal radioactive uptake in bones on the PET images was considered as positive lesion for bone metastasis. The efficacy of 2imaging methods and the supplementary role of CT in the diagnosis of bone metastasis were evaluated by McNemar test. Results:Of 238 patients, 95 were with bone metastases and 143 were without bone metastases, including 436 lesions with bone metastases and 358 lesions without bone metastases. Based on the visual analysis, 18F-FAPI-42 PET showed a higher diagnostic sensitivity than 18F-FDG PET (98.4%(429/436) vs 86.5%(377/436); χ2=41.95, P<0.001), while 18F-FDG PET had a higher diagnostic specificity than 18F-FAPI-42 PET (83.2%(298/358) vs 70.4%(252/358); χ2=22.50, P<0.001), and the accuracies of both methods were similar (85.8%(681/794) vs 85.0%(675/794); χ2=0.16, P=0.685). However, when the positive lesions seen in PET were analyzed combined with the image features on CT by the same scanner, the diagnostic specificity of 18F-FAPI-42 PET/CT was significantly improved compared to that of 18F-FAPI-42 PET alone (91.3%(327/358) vs 70.4%(252/358); χ2=73.01, P<0.001), and was similar to 18F-FDG PET/CT (93.0%(333/358); χ2=0.78, P=0.377). Meanwhile, this combined analysis brought a higher sensitivity and accuracy of 18F-FAPI-42 PET/CT than 18F-FDG PET/CT in diagnosing bone metastases (sensitivity: 98.4%(429/436) vs 86.5%(377/436); χ2=41.95, P<0.001; accuracy: 95.2%(756/794) vs 89.4%(710/794); χ2=21.54, P<0.001). Conclusions:The diagnostic sensitivity of 18F-FAPI-42 PET for bone metastasis is superior to 18F-FDG PET, but the specificity is lower. However, when CT features is combined for analysis, the diagnostic specificity of 18F-FAPI-42 PET/CT is significantly improved, which thus can be used to diagnose bone metastasis accurately and is superior to 18F-FDG PET/CT.

In this study, the pre-market regulatory requirements for typical Class Ⅱ wound dressings, as well as the status of testing and post-market adverse events monitoring, were reviewed from the perspective of the whole lifecycle of medical devices. Additionally, the regulatory requirements for wound dressings in China, the United States, and the European Union were compared. Supplementary research was also conducted on Class Ⅰ and Ⅱ liquid and paste dressing products. Furthermore, this study analyzed the issues in the registration and application of typical Class Ⅱ wound dressings and provided regulatory recommendations, aiming to offer technical references for the review and approval, inspection and testing, and post-market supervision of wound dressing products.
Bandages/standards* ; United States ; China ; Humans ; European Union