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 analyze the ranking of direct economic losses due to postoperative hospital-associated in-fections in patients undergoing surgical excisions for common tumors.METHODS Totally 12,010 tumor patients who were treated in Jiangxi Cancer Hospital from Jan.2019 to Dec.2021 were recruited as the research subjects.The differences in covariates including the population,type of tumor and diagnosis and treatment of tumor without infection between the infection group and the non-infection group were balanced by 1∶1 propensity score matching(PSM).The length of hospital stay and hospitalization costs were observed and compared between the two groups of patients.RESULTS Totally 806 pairs of research subjects were matched,covering 10 sites of tumors(lung,stomach,colon,liver,breast,esophagus,thyroid,cervix,uterine body and brain).In the infec-tion group,the patients with lower respiratory tract infections accounted for 60.52％,the patients with surgical site infections accounted for 27.19％,and the patients with other sites of infections accounted for 12.29％.As compared with the non-infection group,the average length of hospital stay of the infection group was extended by 4 days,and the total hospitalization cost was increased by 9068.99 yuan.Among all the expenses,western medi-cine cost,general treatment operation cost,cost of disposable materials for treatment,cost of laboratory test and nursing cost ranked the top 5 economic losses,and there were significant differences between the two groups(P＜0.05).With the respect to different types of tumors,brain tumors,esophageal tumors,gastric tumors,colorectal tumors and cervical tumors ranked the top 5 economic losses.CONCLUSIONS The tumor patients un-dergoing surgical excisions are at high risk of postoperative hospital-associated infections,which may result in economic losses.It is necessary to focus on the prevention and control of lower respiratory tract infections and SSI.

Objective:To evaluate the efficacy and safety of obinutuzumab combined with glucocorticoid-based therapy in patients with relapsed immune thrombotic thrombocytopenic purpura (iTTP).Methods:This study analyzed the efficacy and adverse reactions of four patients with relapsed iTTP who were treated with a combination of obinutuzumab and glucocorticoids to assess the effectiveness and safety of the treatment.Results:All four patients had a history of multiple relapses and had previously undergone treatment with rituximab and bortezomib. Three patients exhibited additional autoantibodies. Following the combined therapy, all patients achieved clinical remission, with ADAMTS13 activity returning to normal levels and inhibitors testing negative. During a median follow-up period of 11 months (range: 3–17 months), all patients maintained sustained remission. No severe adverse events were reported during treatment or follow-up.Conclusion:The combination of obinutuzumab and glucocorticoid-based therapy is effective and safe for treating relapsed iTTP.

To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

OBJECTIVE To analyze the ranking of direct economic losses due to postoperative hospital-associated in-fections in patients undergoing surgical excisions for common tumors.METHODS Totally 12,010 tumor patients who were treated in Jiangxi Cancer Hospital from Jan.2019 to Dec.2021 were recruited as the research subjects.The differences in covariates including the population,type of tumor and diagnosis and treatment of tumor without infection between the infection group and the non-infection group were balanced by 1∶1 propensity score matching(PSM).The length of hospital stay and hospitalization costs were observed and compared between the two groups of patients.RESULTS Totally 806 pairs of research subjects were matched,covering 10 sites of tumors(lung,stomach,colon,liver,breast,esophagus,thyroid,cervix,uterine body and brain).In the infec-tion group,the patients with lower respiratory tract infections accounted for 60.52％,the patients with surgical site infections accounted for 27.19％,and the patients with other sites of infections accounted for 12.29％.As compared with the non-infection group,the average length of hospital stay of the infection group was extended by 4 days,and the total hospitalization cost was increased by 9068.99 yuan.Among all the expenses,western medi-cine cost,general treatment operation cost,cost of disposable materials for treatment,cost of laboratory test and nursing cost ranked the top 5 economic losses,and there were significant differences between the two groups(P＜0.05).With the respect to different types of tumors,brain tumors,esophageal tumors,gastric tumors,colorectal tumors and cervical tumors ranked the top 5 economic losses.CONCLUSIONS The tumor patients un-dergoing surgical excisions are at high risk of postoperative hospital-associated infections,which may result in economic losses.It is necessary to focus on the prevention and control of lower respiratory tract infections and SSI.

Objective:To evaluate the efficacy and safety of obinutuzumab combined with glucocorticoid-based therapy in patients with relapsed immune thrombotic thrombocytopenic purpura (iTTP).Methods:This study analyzed the efficacy and adverse reactions of four patients with relapsed iTTP who were treated with a combination of obinutuzumab and glucocorticoids to assess the effectiveness and safety of the treatment.Results:All four patients had a history of multiple relapses and had previously undergone treatment with rituximab and bortezomib. Three patients exhibited additional autoantibodies. Following the combined therapy, all patients achieved clinical remission, with ADAMTS13 activity returning to normal levels and inhibitors testing negative. During a median follow-up period of 11 months (range: 3–17 months), all patients maintained sustained remission. No severe adverse events were reported during treatment or follow-up.Conclusion:The combination of obinutuzumab and glucocorticoid-based therapy is effective and safe for treating relapsed iTTP.

Objective:To develop a nomogram predictive model on the basis of identification of the risk factors associated with failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction.Methods:A retrospective study was conducted of the clinical data of the patients who had been treated for dislocation of the subaxial cervical spine with locked facets at Department of Orthopaedic Trauma, The Hospital Affiliated to Guizhou Medical University and Department of Spine Surgery, The People's Hospital of Guizhou Province from January 2014 to December 2022. The clinical data from The Hospital Affiliated to Guizhou Medical University were used as a training set (156 cases) and those from The People's Hospital of Guizhou Province as an external validation set (54 cases). Univariate analysis and multi-variate logistic regression analysis of the training set were conducted to screen out independent risk factors associated with the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. A nomogram predictive model was thus constructed and assessed by the receiver operating characteristic (ROC) curve, calibration curve, and decision curve. Internal validation of the training set and external validation set was used to evaluate and validate the model.Results:The multivariate logistic regression analysis revealed that cervical Ⅰ grade dislocation ( P=0.002), cervical Ⅱ grade dislocation ( P=0.007), low segment affected ( P=0.042), unilateral facet locked ( P=0.027), and the ASIA grading of spinal cord injury ( P=0.008) were the independent risk factors associated with the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction, based on which the nomogram model with a C-index of 0.88 was constructed to predict the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. Analysis of the ROC curve of the training set showed an area under the curve (AUC) of 0.88, indicating good accuracy of the nomogram model. Analysis of the calibration curve showed high consistency between the probability of the nomogram model predicting the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction and the actual probability of traction reposition failure. Analysis of the decision curve showed that application of the nomogram model led to good benefits when the net benefit threshold for the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction was 0.03 to 0.84. Analysis of the ROC curve of external validation set showed an AUC of 0.79, indicating good accuracy of the nomogram model. The training set showed a C-index of 0.87 after 1,000 internal verifications by the Bootstrap method, indicating good discrimination of the nomogram model. Conclusions:Cervical Ⅰ grade dislocation, cervical Ⅱ grade dislocation, low segment affected, unilateral facet locked, and incomplete spinal cord injury are independent risk factors associated with failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. A nomogram model has been successfully constructed which can predict the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. Validation and evaluation of the nomogram model have demonstrated its good predictive value.

As a receptor protein,GPR120 is activated by long-chain fatty acids(such as omega-3 fat-ty acids,alpha-linolenic acid,eicosapentaenoic acid,and docosahexaenoic acid.It plays an important regulatory role in gastrointestinal peptide release,inflammation,lipogenesis,glucose tolerance,and insulin sensitivity.In order to study the synergistic regulation of the GPR120 gene on milk fat me-tabolism and its anti-inflammatory effects in dairy cow MAC-T cells,the GPR120 activator TUG-891 and the inhibitor AH7614 were used to treat both dairy cow MAC-T cells and LTA-induced inflammatory dairy cow MAC-T cells.This treatment aimed to detect the expression of key genes involved in milk fat synthesis and inflammatory factors.The results showed that the GPR120 gene activator significantly increased the relative expression levels of cholesterol regulatory element binding protein(SREBP1)and fatty acid synthase(FASN),key genes for milk fat synthesis.Addi-tionally,the expression levels of the inflammatory factor interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were reduced in the inflammatory dairy cow MAC-T cell model,which prelimi-natively reveals that GPR120 co-regulates milk fat and mammary inflammation in dairy cows,thereby laying a foundation for subsequent molecular mechanism research.

Objective:To review the diagnosis and treatment process of a patient with relapsed immune thrombotic thrombocytopenic purpura (iTTP) secondary to systemic lupus erythematosus (SLE) and conduct a review of relevant literature, so as to provide a reference basis for the diagnosis and treatment of this disease.Methods:We retrospectively analyzed the diagnosis and treatment process of a patient with relapsed iTTP secondary to SLE who was admitted to the First Affiliated Hospital of Soochow University, and conducted a review of literature.Results:When the patient's iTTP relapsed, it was accompanied by moderate lupus activity, hypofibrinogenemia, renal insufficiency, and pulmonary infection. After adopting therapeutic plasma exchange (TPE), hormone pulse therapy, bortezomib, and comprehensive diagnosis and treatment by a multidisciplinary team, both the patient's iTTP and SLE were relieved.Conclusion:A comprehensive examination should be completed when diagnosing iTTP to rule out the possibility of combining with other autoimmune diseases. For relapsed and refractory cases, treatments targeting plasma cells, such as bortezomib, can be attempted.