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.

This study aims to analyze the international research trends and frontier hotspots related to the application of growth mindset in medical education from 2016 to 2024. The objective is to provide insights and references for the development of relevant research in China.

Objective: To evaluate the clinical feasibility and potential for malignant transformation prevention of hyperkeratotic oral leukoplakia (OLK) treatment using erbium, chromium:yttrium-scandium-gallium-garnet laser (Er,Cr:YSGG) ablation combined with 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT), providing a reference for clinical practice. Methods: Two cases of histopathologically confirmed gingival leukoplakia were treated. Following Er,Cr:YSGG (wavelength: 2,780 nm) ablation of the hyperkeratotic layer, 20% ALA solution was topically applied for 3 hours, followed by irradiation with a 635 nm diode laser (energy density: 100 J/cm²). Follow-up assessments included lesion regression and exfoliative cytology DNA ploidy stability, supplemented by a systematic literature review on OLK treatment advancements. Results: Case 1: at 1-year follow-up, > 90% lesion regression was observed (residual: 2 mm circular white patch) without recurrence, with stable diploid DNA ploidy. Case 2: complete lesion resolution and no recurrence at 1 year, with stable diploid DNA ploidy. Literature review showed that pharmacological therapies ameliorate OLK symptoms but fail to halt malignant progression, while surgical interventions carry recurrence risks. PDT demonstrates unique advantages by selectively targeting dysplastic cells. However, uneven ALA penetration due to the keratin barrier limits efficacy. Compared to needling, diode laser, or CO₂ laser pretreatment, Er,Cr:YSGG leverages water absorption for precise, low-thermal keratin ablation, substantially enhancing tissue permeability and optimizing ALA diffusion, thereby improving PDT outcomes. Conclusion Er,Cr:YSGG ablation combined with PDT serves as an optimized regimen for hyperkeratotic OLK, offering a novel approach to mitigate OLK carcinogenesis.

According the Good Clinical Practice(GCP)and programmatic requirements,we analyze the management characteristics and the costs of drugs for clinical trials in different specialties from the drug management;The characteristics of the management of drugs for different specialties was summarize and the differential factors that may affect the management cost was explored,so as to provide theoretical support for the research institutions to utilize the resources in a rational and efficient way.This article provides a guarantee for the drug management with the aim of enhancing the quality and efficiency of clinical trials.

Objective To investigate the expression and significance of RANK/RANKL/OPG signaling path-way in gerbil external auditory canal cholesteatoma(EACC).Methods Twenty-five mongolian long-clawed gerbils were divided into an experimental group(n=15)and a control group(n=10),and an EACC model was constructed by external auditory canal ligation method in the experimental group.Three months later,the external auditory ca-nal tissues of the control group and the experimental group(10 ears)were collected,and RT-qPCR and Western blot were performed to detect the expression levels of receptor activator of NF-κB(RANK),receptor activator of NF-κB ligand(RANKL),osteoprotegerin(OPG)mRNA and protein in the signaling pathway.Results Two ger-bils in the experimental group died during the modeling process,and cholesteatoma of the external auditory canal was formed in 10 ears 3 months after the operation,with a modeling success rate of 76.9％(10/13).Compared with the normal control group,there was no statistically significant difference in the expression of RANK mRNA in gerbil external auditory canal tissues(P=0.08).The expression of RANK protein was upregulated,the expression of RANKL mRNA and protein was upregulated,while the expression of OPG mRNA and protein was downregulat-ed,the ratio of RANKL/OPG was upregulated,with a statistically significant difference between the two groups(P＜0.01).Conclusion The RANK/RANKL/OPG signaling system may play an important role in the formation of gerbil EACC and bone destruction.

Objective:To observe the changes in serum estradiol (E 2) levels in pregnant women of different gestational weeks and their predictive value for early intrauterine pregnancy outcomes. Methods:A retrospective study was conducted involving 375 pregnant women who were treated at the Affiliated Yangming Hospital of Ningbo University (Yuyao People's Hospital) from September 2021 to September 2023. The clinical data were categorized based on pregnancy outcomes into three groups: a normal pregnancy group ( n = 150), a threatened miscarriage with continued pregnancy group ( n = 150), and a miscarriage group ( n = 75). The serum estradiol (E 2) levels at different gestational weeks were compared among the three groups: 5 to < 6 weeks (35-41 days), 6 to < 7 weeks (42-48 days), and 7 to 8 weeks (49-55 days). The predictive value of serum E 2 levels for early intrauterine pregnancy outcomes across different gestational weeks was analyzed using Receiver Operating Characteristic (ROC) curves. Results:In the normal pregnancy group, the serum E 2 levels at different gestational weeks were as follows: (1 691.87 ± 532.21) pmol/L for 5 to < 6 weeks, (2 376.64 ± 788.36) pmol/L for 6 to < 7 weeks, and (3 576.30 ± 1,190.06) pmol/L for 7 to 8 weeks. These values were significantly higher than those in the threatened miscarriage with continued pregnancy group [(1 409.28 ± 473.49) pmol/L, (1 893.13 ± 563.15) pmol/L, (2 035.79 ± 612.47) pmol/L, t = 5.15, 11.68, 6.60, all P < 0.05] and the miscarriage group [(906.49 ± 338.09) pmol/L, (923.63 ± 365.39) pmol/L, (950.27 ± 378.89) pmol/L, t = 16.19, 15.45, 21.50, all P < 0.05]. The serum E 2 levels at different gestational weeks in the threatened miscarriage with continued pregnancy group were significantly higher than those in the miscarriage group ( t = 7.48, 10.81, 8.89, all P < 0.05). Both the normal pregnancy group and the threatened miscarriage with continued pregnancy group showed an increasing trend in serum E 2 levels with advancing gestational weeks ( t = 6.74, 18.55, 7.58, 9.82, 11.81, 2.24, all P < 0.05). In contrast, the serum E 2 levels in the miscarriage group also increased with advancing gestational weeks, but the differences were not statistically significant ( P > 0.05). The results from the receiver operating characteristic curve analysis indicated that the areas under the curve for predicting early intrauterine pregnancy outcomes based on serum E 2 levels at different gestational weeks were 0.857, 0.810, and 0.839, demonstrating excellent diagnostic efficacy. Conclusions:Dynamic monitoring of serum E 2 levels is beneficial for predicting early intrauterine pregnancy outcomes and providing guidance for clinical diagnosis and treatment.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Severe open tibiofibular fractures account for approximately 28.1% of all open fractures. Among them, Gustilo-Anderson type IIIB/C fractures present significant clinical challenges due to associated bone and soft tissue defects, high infection rates, and risk of amputation. Inadequate preoperative assessment may lead to suboptimal emergency surgical planning or intraoperative complications. Historically, external fixation was often preferred, but this approach has been associated with limitations such as restricted joint mobility, delayed bone union, joint stiffness, and disuse osteoporosis, resulting in poor functional recovery. With advancements of debridement techniques, standardization of antibiotic use, and popularization of early soft tissue coverage, early internal fixation has gained broader acceptance. Nevertheless, controversies persist regarding the choice of fixation method, timing of definitive fixation, use of reamed versus unreamed intramedullary nailing, and necessity of fibular fixation. To standardize the diagnosis and early management of severe open tibiofibular fractures, reduce complication rates, and improve functional recovery, the Society of Microsurgery of the Chinese Medical Association organized a panel of domestic experts to develop the Evidence-based guideline for the diagnosis and early fixation of severe open tibiofibular fractures ( version 2025), using evidence-based methodology. The guidelines provided 12 recommendations covering diagnostic and early fixation strategies of severe open tibiofibular fractures, aiming to provide clinicians with scientifically grounded and standardized guidance.

Objective:To assess the implementation status of Internet-Plus Nursing Services from patient experience perspectives and analyze influencing factors in Putuo District, Shanghai.Methods:A cross-sectional study on Internet-Plus Nursing Services was conducted from January to March 2024. The study enrolled 502 patients from 12 community health centers and 4 social nursing stations in Shanghai Putou District via convenience sampling. Using Internet-Plus Nursing Services for ≥ 6 months were defined as having received the services. Service implementation and quality perception were evaluated using satisfaction assessments and SERVQUAL-based quality perception scales. Data analysis included descriptive statistics and binary logistic regression.Results:The service adoption rate was 73.71% (370/502), with 66.93%(336/502) policy awareness. The logistic regression analysis indicated that the application of services was significantly influenced by age ( OR=0.660, 95% CI: 0.512-0.850), self-care ability ( OR=0.384, 95% CI: 0.257-0.574), and policy awareness ( OR=0.344, 95% CI:0.210-0.564) (all P<0.05). Among the service types, specimen collection/monitoring (80.27%, 299/370) and home health guidance (64.86%,240/370) were most utilized. Injection services had the highest satisfaction (4.10/5), while the special type of care had the lowest satisfaction rate (2.45/5). Quality perception was highest for Assurance and Empathy dimensions (both 4.74/5). Conclusion:The application rate and satisfaction of Internet-Plus Nursing Service in Shanghai Putuo District is high, which is associated with effective integration of clinical needs and perceptual drivers. Targeted expansion of specialized care services is recommended to enhance service diversification.