Objective To investigate the causes and management strategies for lower limb ischemic necrosis following xenogeneic heterotopic heart transplantation from a multigene-edited pig to a rhesus monkey. Methods A xenogeneic heterotopic heart transplantation was performed on December 16, 2023, at the Institute of Experimental Animals of Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, using a quintuple-gene-edited pig as the donor and a rhesus monkey as the recipient. On postoperative day (POD) 9, the recipient monkey underwent left lower limb amputation due to ischemic necrosis. Blood samples were collected at various time points after transplantation for analysis of hematologic parameters, liver and renal function, myocardial enzymes, and coagulation profiles. Ultrasound and computed tomography (CT) were used to evaluate anastomotic patency and cardiac structure. Immunological assays, including complement-dependent cytotoxicity (CDC) and IgG/IgM antibody detection, combined with clinical observations, were employed to assess rejection type and therapeutic response. Results The recipient monkey survived for 46 days after transplantation. Echocardiography demonstrated preserved biventricular systolic function in the recipient’s native heart, with left ventricular ejection fraction (LVEF) consistently exceeding 50%. In the donor pig heart, left ventricular endocardial thickening was noted on POD 9, followed by right ventricular endocardial thickening on POD 24, while LVEF remained around 35%. No hyperacute or acute rejection was detected immunologically. CDC positivity ranged between 3.4% and 5.1%, with IgG/IgM antibody binding trends consistent with CDC results. Following amputation, the recipient exhibited elevated inflammatory markers, coagulopathy, and reactive thrombocytosis, which later normalized. Immunohistochemical staining of the necrotic limb revealed arterial and venous thrombosis; however, no T-cell or B-cell infiltration was observed in vascular structures, thrombi, nerves, muscles, fascia, or skin tissues, with CD3 and CD20 staining both negative. Conclusion Limb ischemia after xenogeneic heart transplantation may be associated with lower extremity vascular thrombosis triggered by local trauma in the context of transplantation-induced inflammatory activation and coagulation dysfunction. While no clear lymphocyte-mediated rejection was observed, further studies are needed to explore the potential role of non-lymphocyte-mediated immune mechanisms.

Objective To evaluate the impact of incorporating two doses of varicella vaccine into the immunization program in Shanghai on the incidence of varicella, age-specific incidence, and varicella outbreaks in kindergartens and schools in Jinshan District, and to assess its prevention and control effects. Methods Descriptive epidemiological methods were used to analyze and compare the population distribution characteristics and the occurrence of varicella outbreaks before (2013-2017) and after (2018-2024) the implementation of the strategy. Interrupted time series (ITS) design and its segmented regression model were used to analyze the changes in the incidence of varicella in the two stages before and after the implementation of the vaccination of two doses of varicella vaccine, and to evaluate the short-term and long-term preventive and control effects of the strategy. Results After the incorporation of two doses of varicella vaccine into the immunization program, the average annual incidence of varicella in Jinshan District decreased from 82.19/100 000 (2013-2017) to 53.51/100 000 (2018-2024). The interrupted time series analysis showed that after the implementation of the strategy, the slope change in the incidence of varicella was -10.20/100 000 (t=-2.38, P=0.04), and the overall trend was downward (slope β₁+β₃= -8.34/100 000). Except for the 30-49 years old group, the incidence rates in all other age groups decreased to varying degrees. Among them, the 5-9 years old group had a better vaccine efficacy (VE) of 52.23% (95% CI: 49.07-60.65), followed by the 10-14 years old group. The number of potential varicella outbreaks and cluster outbreaks in nurseries, kindergartens, and schools across the district dropped significantly. Conclusion Incorporating two doses of varicella vaccine into the immunization program can effectively reduce the incidence of varicella and the number of varicella outbreaks in kindergartens and schools, which is of great significance for the prevention and control of varicella.

Objective: To analyze the change trends in the body shape indicators and proportions of students in Harbin from 2010 to 2024, and to investigate ergonomic compatibility of functional dimensions of school desks and chairs with current student shape indicators, so as to provide a reference for revising furniture standards of desks and chairs. Methods: Between September and November of both 2010 and 2024, a combination of convenience sampling and stratified cluster random sampling was conducted across three districts in Harbin, yielding samples of 6 590 and 6 252 students, respectively. Anthropometric shape indicators cluding height, sitting height, crus length, and thigh length-and their proportional changes were compared over the 15-year period. The 2024 data were compared with current standard functional dimensions of school furniture. The statistical analysis incorporated t-test and Mann-Whitney U- test. Results: From 2010 to 2024, average height increased by 1.8 cm for boys and 1.5 cm for girls; sitting height increased by 1.5 cm for both genders; crus length increased by 0.3 cm for boys and 0.4 cm for girls; and thigh length increased by 0.5 cm for both genders. The ratios of sitting height to height, and sitting height to leg length increased by less than 0.1 . The difference between desk chair height and 1/3 sitting height ranged from 0.4-0.8 cm. Among students matched with size 0 desks and chairs, 22.0% had a desk to chair height difference less than 0, indicating that the desk to chair height difference might be insufficient for taller students. The differences between seat height and fibular height ranged from -1.4 to 1.1 cm; and the differences between seat depth and buttock popliteal length ranged from -9.8 to 3.4 cm. Among obese students, the differences between seat width and 1/2 hip circumference ranged from -20.5 to -8.7 cm, while it ranged from -12.2 to -3.8 cm among non obese students. Conclusion Current furniture standards basically satisfy hygienic requirements; however, in the case of exceptionally tall and obese students, ergonomic accommodations such as adaptive seating allocation or personalized adjustments are recommended to meet hygienic requirements.

Although clinical evidence suggests that nonalcoholic fatty liver disease is an established major risk factor for heart failure, it remains unexplored whether sleep disorder-caused hepatic damage contributes to the development of cardiovascular disease (CVD). Here, our findings revealed that sleep fragmentation (SF) displayed notable hepatic detrimental phenotypes, including steatosis and oxidative damage, along with significant abnormalities in cardiac structure and function. All these pathological changes persisted even after sleep recovery for 2 consecutive weeks or more, displaying memory properties. Mechanistically, persistent higher expression of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in the liver was the key initiator of SF-accelerated damage phenotypes. SF epigenetically controlled the acetylation of histone H3 lysine 27 (H3K27ac) enrichment at the Nox4 promoter and markedly increased Nox4 expression in liver even after sleep recovery. Moreover, fine coordination of the circadian clock and hepatic damage was strictly controlled by BMAL1-dependent Sirtuin 1 (Sirt1) transcription after circadian misalignment. Accordingly, genetic manipulation of liver-specific Nox4 or Sirt1, along with pharmacological intervention targeting NOX4 (GLX351322) or SIRT1 (Resveratrol), could effectively erase the epigenetic modification of Nox4 by reducing the H3K27ac level and ameliorate the progression of liver pathology, thereby counteracting SF-evoked sustained CVD. Collectively, our findings may pave the way for strategies to mitigate myocardial injury from persistent hepatic detrimental memory in diabetic patients.

G protein-coupled receptors (GPCRs) are significant drug targets, but their potential in cancer therapy remains underexplored. Conventional GPCR agonists or antagonists have shown limited effectiveness in cancer treatment, necessitating new GPCR-targeting strategies for more effective therapies. This study discovers that Yersinia pestis LcrV, a crucial linker protein for plague infection, acts as a biased agonist of a GPCR, the formyl peptide receptor 1 (FPR1). The LcrV protein induces unique conformational changes in FPR1, resulting in G proteins being activated in a distinctive state without subunit dissociation. This leads to a biased signaling profile characterized by cyclic adenosine monophosphate (cAMP) responses and β-arrestin2 recruitment, but not calcium mobilization. In FPR1-expressing triple-negative breast cancer (TNBC) cells, LcrV bi-directionally modulates intracellular signaling pathways, downregulating extracellular signal-regulated kinases (ERK1/2) and Akt pathways while upregulating Jun N-terminal kinase (JNK) and p38 pathways. This dual modulation results in cell cycle arrest and the inhibition of TNBC cell proliferation. In TNBC xenograft mouse models, long-term LcrV treatment inhibits tumor growth more effectively than a conventional FPR1 antagonist. Additionally, LcrV treatment reprograms tumor cells by reducing stemness-associated proteins OCT4 and c-MYC. Our findings highlight the potential of biased GPCR agonists as a novel GPCR-targeting strategy for cancer treatment.

Objective:To evaluate the safety and effectiveness of fuzuloparib for the treatment of ovarian epithelial cancer patients in the real world setting.Methods:A retrospective analysis was conducted on the baseline data of 4 620 ovarian cancer patients who had received fuzuloparib monotherapy or combination therapy. Another 224 ovarian cancer patients who were willing to receive fuzuloparib monotherapy or combination therapy were prospectively enrolled, and their baseline characteristics, drug effectiveness, and safety data were analyzed.Results:(1) Among the 4 620 patients in the retrospective cohort, the median age of patients was 60 years; tumor types: 89.8% (4 149/4 620) had ovarian cancer. Among patients with clearly documented information, the vast majority had a histological type of serous carcinoma (82.9%, 3 770/4 546) and International Federation of Gynecology and Obstetrics (FIGO) staging of Ⅲ-Ⅳ (90.9%, 1 537/1 691). (2) Among the 224 patients in the prospective cohort, the median age of patients was 57 years; tumor types: 83.9% (188/224) had ovarian cancer. Among patients with clearly documented records, the predominant pathologic type was serous carcinoma (91.9%, 193/210), and FIGO stage was Ⅲ-Ⅳ in 79.9% (139/174). (3) Among the 224 prospective patients: 84 patients received first-line fluzoparib maintenance therapy, 92 patients received fluzoparib maintenance therapy after platinum-sensitive recurrence, 23 patients received direct fluzoparib treatment after platinum-sensitive recurrence, 19 patients received direct fluzoparib treatment after platinum-resistant recurrence. The median follow-up durations were 8.5, 8.7, 7.9, and 6.7 months, respectively. The median durations of fluzoparib treatment were 6.7, 4.8, 3.1, and 1.9 months, respectively. The median progression-free survival (PFS) times were not reached during follow-up, 12.6 months, not reached during follow-up, and 4.8 months, respectively. The 1-year PFS rates were 84.1%, 55.0%, 69.8%, and 45.5%, respectively. The remaining 6 patients received other fluzoparib regimens. (4) Among the 224 patients in the prospective dataset, 205 had safety data recorded. Of these, 127 patients (62.0%, 127/205) experienced treatment-related adverse events, with common events including anemia (24.4%, 50/205), thrombocytopenia (21.0%, 43/205), and leukopenia (19.5%, 40/205). Among the 205 patients, 43 (21.0%, 43/205) experienced grade 3 or higher treatment-related adverse events, with common events including anemia (8.3%, 17/205) and thrombocytopenia (8.3%, 17/205).Conclusions:The effectiveness of fuzuloparib in clinical application is generally consistent with other drugs in the same class, with good safety. This study provids new clinical evidence for the treatment of ovarian cancer with fuzuloparib.

Objective:To investigate the effect of basket assisted retrograde ureteral soft lithotripsis retrograde intrarenal surgery (RIRS) in the treatment of subcalyceal calculi and its effect on stone clearance.Methods:A retrospective cohort study method was conducted on 120 patients with lower calyx stones treated at Jiaozhou Central Hospital of Qingdao City, from October 2021 to January 2024. There were 66 males and 54 females, aged (48.10±10.11) years old. The patients were divided into two groups based on their surgical methods: 60 patients who received only RIRS treatment were in the control group, while 60 patients who underwent RIRS with the assistance of a stone retrieval basket were in the observation group. The study compared various perioperative indicators (such as intraoperative blood loss, surgical duration, and hospital stay), as well as the stone clearance rates, inflammation levels, and postoperative complications at 3 days (short-term) and 30 days (long-term) after surgery. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was employed for inter-group comparisons. Chi-square test was used for inter-group comparisons of count data. Results:The operation time of the observation group was (78.26±10.54) min, and that of the control group was (70.65±8.67) min. The difference between the two groups was statistically significant ( P<0.05). There was no difference between the two groups in intraoperative bleeding, the first time to get out of bed after surgery and the postoperative hospital stay ( P>0.05). The stone expulsion time in the observation group was (16.38±4.22) d, while that in the control group was (18.96±5.10) d, the stone removal rates of the observation group at 3 days and 30 days after surgery were 88.33% and 95.00% respectively, while those of the control group were 73.33% and 83.33% respectively. There was a statistically significant difference between the two groups ( P<0.05). The stone expulsion time was shorter in the observation group compared to the control group, with a statistically significant difference ( P<0.05). There were no significant differences in postoperative inflammatory factors between the two groups( P>0.05). The incidence of postoperative complications in the observation group was 6.67%, and that in the control group was 20.00%. The difference between the two groups was statistically significant ( P<0.05). Conclusions:Stone basket removal combined with RIRS is safe and feasible, which can not only improve the stone clearance rate, but also reduce the incidence of postoperative complications without increasing the inflammatory reaction, and has a significant effect. However, the clinical application needs to strictly grasp the indications and avoid overtreatment in combination with the individual situation of the patients.

Objective:To investigate the clinical demand for subcutaneous immunotherapy (SCIT) with pet allergens and explore the sensitization characteristics of patients undergoing pet SCIT.Methods:A cross-sectional retrospective analysis was conducted on patients diagnosed with pet allergies and treated with pet allergen SCIT in our outpatient clinic from January 2019 to December 2023. Patients were categorized into three groups based on the type of SCIT received: single-cat SCIT group, single-dog SCIT group, and combined cat-dog SCIT group.Results:A total of 931 patients were included, the age range was 5-65 years (median age of 30 years), with 283 male and 648 female. Among them, 67.7%( n=630) received single-cat SCIT, 10.9% ( n=102)received single-dog SCIT, and 21.4% ( n=199) received combined cat-dog SCIT. The number of patients receiving pet allergen SCIT increased annually. Patients in the single-dog SCIT group were significantly older than those in the other two groups ( H=41.329, P<0.001) and had a lower prevalence of allergic rhinitis (91.2% vs. 96.5% and 98.5%; χ2=10.400, P=0.006). In the combined cat-dog SCIT group, the allergy rate to mold allergens was significantly higher than in the single-cat SCIT group (12.6% vs. 4.9%, χ2=13.965, P=0.001). In the single-dog SCIT group, the allergy rate to spring pollen allergens was significantly higher than in the other two groups ( χ2=15.731, P<0.001), and the allergy rate to autumn pollen allergens was significantly higher than in the single-cat SCIT group ( χ2=13.459, P=0.001). There was no significant difference in the dust mite allergy rate among the three groups( χ2=4.117, P=0.129). In the single-dog SCIT group, patients with asthma were significantly older than those without asthma (41.2 vs. 35.2 years old, t=-2.073, P=0.041). In both the single-cat and single-dog SCIT groups, the proportion of allergic rhinitis in the asthma group(91.2%,78.3%) was significantly lower than that in the non-asthma group(97.4%,94.9%) ( χ2=8.863,6.158; P=0.008,0.026). In the single-cat SCIT group, non-asthmatic patients were significantly more likely to receive SCIT combined with spring pollen allergens compared to asthmatic patients (23.9% vs. 11.0%, χ2=7.586, P=0.006). Conclusions:The demand for pet allergen SCIT has steadily increased over the years, with a predominance of female patients. Sensitization profiles varied among patients receiving SCIT for different pet allergens. This study comprehensively elucidates the clinical demand and sensitization characteristics of patients undergoing pet allergen SCIT, providing valuable reference data for clinical diagnosis and treatment.

Objective:To explore the activation pathway of CD4+CD28-T cells and their specific role in rheumatoid arthritis(RA).Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from RA patients and healthy individuals.Flow cytome-try was used to detect the proportion and activation level of CD4+CD28-T cells,as well as the expressions of perforin and TNF-α,and to analyze the correlation between the proportion of CD4+CD28-T cells and age,C-reactive protein,anti-cyclic citrullinated peptide(CCP)antibodies,rheumatoid factors(RF)and erythrocyte sedimentation rate(ESR).The expression of Toll-like receptor(TLR)2 on CD4+CD28-T cells was detected by flow cytometry.Purified RA CD4+T cells were stimulated with anti-CD3 antibody and TLR2 agonist alone or in combination,and the activation level and cytokine secretion of CD4+CD28-T cells were detected by flow cytometry.Results:The proportion of CD4+CD28-T cells in RA patients was higher than that in healthy individuals(P<0.05),and was positively correlated with rheumatoid factors and erythrocyte sedimentation rate(r=0.554 1,P<0.01;r=0.363 4,P<0.01).Compared with CD4+CD28+T cells,CD25 expression was downregulate(P<0.01),but CD40L and TLR2 expressions were upregulated(P<0.01;P<0.000 1)in CD4+CD28-T cells from RA patients,and they secreted more perforin and TNF-α(P<0.001;P<0.001).After TLR2 ago-nist stimulation,there was no significant change in CD40L and CD25 expression,as well as perforin and TNF-α secretion in CD4+CD28-T cells from RA patients.Conclusion:The proportion of CD4+CD28-T cells is increased in RA patients,and they secrete a large amount of perforin and TNF-α,which may be involved in the inflammatory state and joint damage of RA,but TLR2 cannot re-place CD28 in activating CD4+CD28-T cells.

Objective:To explore the activation pathway of CD4+CD28-T cells and their specific role in rheumatoid arthritis(RA).Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from RA patients and healthy individuals.Flow cytome-try was used to detect the proportion and activation level of CD4+CD28-T cells,as well as the expressions of perforin and TNF-α,and to analyze the correlation between the proportion of CD4+CD28-T cells and age,C-reactive protein,anti-cyclic citrullinated peptide(CCP)antibodies,rheumatoid factors(RF)and erythrocyte sedimentation rate(ESR).The expression of Toll-like receptor(TLR)2 on CD4+CD28-T cells was detected by flow cytometry.Purified RA CD4+T cells were stimulated with anti-CD3 antibody and TLR2 agonist alone or in combination,and the activation level and cytokine secretion of CD4+CD28-T cells were detected by flow cytometry.Results:The proportion of CD4+CD28-T cells in RA patients was higher than that in healthy individuals(P<0.05),and was positively correlated with rheumatoid factors and erythrocyte sedimentation rate(r=0.554 1,P<0.01;r=0.363 4,P<0.01).Compared with CD4+CD28+T cells,CD25 expression was downregulate(P<0.01),but CD40L and TLR2 expressions were upregulated(P<0.01;P<0.000 1)in CD4+CD28-T cells from RA patients,and they secreted more perforin and TNF-α(P<0.001;P<0.001).After TLR2 ago-nist stimulation,there was no significant change in CD40L and CD25 expression,as well as perforin and TNF-α secretion in CD4+CD28-T cells from RA patients.Conclusion:The proportion of CD4+CD28-T cells is increased in RA patients,and they secrete a large amount of perforin and TNF-α,which may be involved in the inflammatory state and joint damage of RA,but TLR2 cannot re-place CD28 in activating CD4+CD28-T cells.