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 investigate the changing trends in cardiac function following xenogeneic heterotopic heart transplantation of multi-gene edited pig hearts and assess the impact of recipient immune responses on donor heart, laying experimental groundwork for the clinical application of gene editing technology.Methods:On December 16, 2023, xenogeneic heterotopic heart transplantation was performed between pigs and rhesus monkeys. Functional status of the graft under post-transplantation load conditions and recipient immune indicators were observed.Results:The recipient monkeys survived for 40 days with satisfactory functionality of both donor and recipient hearts, and no hyperacute or acute immune rejection reactions were observed.Conclusion:Multi-gene editing technology provides potential for xenotransplantation, yet further exploration is needed for its clinical application.

Objective To analyze the treatment effects of extracorporeal cardiopulmonary resuscitation(ECPR)in 3 patients with cardiac arrest due to cardiac causes in a mountain area hospital,providing a reference for clinical diagnosis and treatment.Methods The combined treatment process of 3 patients with cardiogenic cardiac arrest admitted to Wufeng Tujia Autonomous County People's Hospital from January to December 2023 was retrospectively analyzed,and the treatment experience was summarized.Results All 3 patients underwent continuous cardiopulmonary resuscitation and defibrillation while urgently initiating the ECPR combined rescue process.First,a central venous catheter(CVC)was established under ultrasound guidance to create access for extracorporeal membrane oxygenation(ECMO).Once the ECMO equipment from a higher-level hospital arrived,the circuit was replaced,significantly reducing the time required for subsequent patient treatment.Finally,two patients with acute myocardial infarction underwent emergency coronary angiography and percutaneous coronary intervention(PCI)under ECMO support,resulting in the restoration of spontaneous cardiac rhythm and gradual stabilization of vital signs.The last patient was discharged after recovery following transfer to a higher-level hospital,while another patient received rehabilitation treatment after hemodynamic stability was achieved.The third patient,who experienced cardiac arrest due to hypertrophic cardiomyopathy,regained spontaneous rhythm after ECMO,but due to poor neurological recovery after transfer to a higher-level hospital,the family chose to withdraw treatment.Conclusions ECPR is a rapid extracorporeal membrane oxygenation-assisted cardiopulmonary resuscitation method for patients who cannot regain spontaneous rhythm or experience recurrent cardiac arrest.It aims to improve patient survival rates.In grassroots medical centers lacking the necessary conditions,early assistance from regional advanced medical centers can ensure rapid transfer and surgery under ECMO support,providing a guarantee for favorable patient outcomes.

Objective To analyze the treatment effects of extracorporeal cardiopulmonary resuscitation(ECPR)in 3 patients with cardiac arrest due to cardiac causes in a mountain area hospital,providing a reference for clinical diagnosis and treatment.Methods The combined treatment process of 3 patients with cardiogenic cardiac arrest admitted to Wufeng Tujia Autonomous County People's Hospital from January to December 2023 was retrospectively analyzed,and the treatment experience was summarized.Results All 3 patients underwent continuous cardiopulmonary resuscitation and defibrillation while urgently initiating the ECPR combined rescue process.First,a central venous catheter(CVC)was established under ultrasound guidance to create access for extracorporeal membrane oxygenation(ECMO).Once the ECMO equipment from a higher-level hospital arrived,the circuit was replaced,significantly reducing the time required for subsequent patient treatment.Finally,two patients with acute myocardial infarction underwent emergency coronary angiography and percutaneous coronary intervention(PCI)under ECMO support,resulting in the restoration of spontaneous cardiac rhythm and gradual stabilization of vital signs.The last patient was discharged after recovery following transfer to a higher-level hospital,while another patient received rehabilitation treatment after hemodynamic stability was achieved.The third patient,who experienced cardiac arrest due to hypertrophic cardiomyopathy,regained spontaneous rhythm after ECMO,but due to poor neurological recovery after transfer to a higher-level hospital,the family chose to withdraw treatment.Conclusions ECPR is a rapid extracorporeal membrane oxygenation-assisted cardiopulmonary resuscitation method for patients who cannot regain spontaneous rhythm or experience recurrent cardiac arrest.It aims to improve patient survival rates.In grassroots medical centers lacking the necessary conditions,early assistance from regional advanced medical centers can ensure rapid transfer and surgery under ECMO support,providing a guarantee for favorable patient outcomes.

BACKGROUND: Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) are the main causes of restenosis (RS) in diabetic lower extremity arterial disease (LEAD). However, the relevant pathogenic mechanisms are poorly understood. METHODS: In this study, we introduced a "two-step injury protocol" rat RS model, which started with the induction of atherosclerosis (AS) and was followed by percutaneous transluminal angioplasty (PTA). Hematoxylin-eosin (HE) staining and immunohistochemistry staining were used to verify the form of RS. Two-step transfection was performed, with the first transfection of Lin28a followed by a second transfection of let-7c and let-7g, to explore the possible mechanism by which Lin28a exerted effects. 5-ethynyl-2΄-deoxyuridine (EdU) and Transwell assay were performed to evaluate the ability of proliferation and migration of VSMCs. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect the expression of Lin28a protein and let-7 family members. RESULTS: Using a combination of in vitro and in vivo experiments, we discovered that let-7c, let-7g, and microRNA98 (miR98) were downstream targets of Lin28a. More importantly, decreased expression of let-7c/let-7g increased Lin28a, leading to further inhibition of let-7c/let-7g. We also found an increased level of let-7d in the RS pathological condition, suggesting that it may function as a protective regulator of the Lin28a/let-7 loop by inhibiting the proliferation and migration of VSMCs. CONCLUSION These findings indicated the presence of a double-negative feedback loop consisting of Lin28a and let-7c/let-7g, which may be responsible for the vicious behavior of VSMCs in RS.
Rats ; Animals ; Down-Regulation ; MicroRNAs/metabolism* ; Feedback ; Cell Proliferation/genetics* ; Atherosclerosis

OBJECTIVE: The current fertilization methods for Chinese yam are uneconomic and unfriend to environment. A rational one is very important to achieve desired balance of high yield of Chinese yam, economic and friend to environment. Here, we studied the effects of nitrogen (N), phosphorus (P), and potassium (K) fertilizers on the yield of 'Qinfeng' Chinese yam in shallow-groove directional cultivation. METHODS: The experiments were conducted in Dehua County, Fujian Province, China using a "3414" optimal design. Overall, three fertilizer factors (N, P, and K) were evaluated at the following four levels: 0, no fertilizer; 1, 0.5-fold the typical rate; 2, typical fertilization rate; and 3, 1.5-fold the typical rate. There were 14 different fertilization treatments. RESULTS: Treatment 6 (N₂P₂K₂) produced the longest (75.6 cm) and thickest tubers (4.9 cm) with the highest tuber fresh weight (1311.9 g) and yield (41 015.9 kg/hm²), whereas, treatment 1 produced the shortest (65.6 cm) and thinnest tubers (3.9 cm) with the lowest fresh weight (953.4 g) and yield (28 532.8 kg/hm²) among the 14 fertilizer combinations. The experimental data could be fitted to single-variable quadratic and binary quadratic models but not to a ternary quadratic polynomial model. Appropriate N, P, and K fertilizer application rates increased Chinese yam yield. However, excessive fertilization lowered the yield. Chinese yam yield was significantly and strongly correlated with the amounts of N, P, and K fertilizer applied. CONCLUSION Based on the single variable quadratic and binary quadratic models, we propose that the quantities of N, P, and K fertilizer used to grow 1 hm² 'Qinfeng' Chinese yam should be 360-388.3, 90-100.95, and 416.3-675 kg, respectively.

Objective:To investigate the effect of peripheral blood neutrophil to lymphocyte ratio (NLR) and platelet distribution width (PDW) on the clinical prognosis of childhood medulloblastomaMethods:Clinical data and survival data of 76 pediatric patients who were diagnosed as medulloblastoma by post-operative pathology in the First Affiliated Hospital of Zhengzhou University from January 2009 to December 2016 were collected. Kaplan- Meier method was used to calculate the overall survival(OS) and progression free survival(EFS) rates, Log- rank test was employed to compare the survival rates of different groups, and Cox proportional hazards regression model was used for multivariate analysis. Results:The log- rank test revealed that 5-year PFS rate and OS rate (22.2%, 22.2%) in the high NLR group (NLR>4.94) were significantly lower than those in the low NLR group (NLR≤4.94) (45.6%, 55.7%), and the differences were statistically significant(PFS: P=0.009, OS: P=0.001), and the 5-year PFS and OS (52.3%, 66.5%) of the high PDW group (PDW>15.90) were significantly higher than those in the low PDW group (PDW ≤ 15.90) (27.1%, 32.5%), and the differences were statistically significant(PFS: P=0.032, OS: P=0.039). Univa-riate analysis showed that the extent of resection (PFS: P=0.006, OS: P=0.009), and postoperative radiotherapy (PFS: P=0.011, OS: P=0.001) and postoperative radiotherapy(PFS: P=0.011, OS: P=0.001) were the factors influencing the prognosis of children with medulloblastoma.Multivariate Cox proportional hazards regression model analysis suggested that no postoperative radiotherapy (PFS: P=0.048, OS: P=0.008), NLR>4.94 (PFS: P=0.023, OS: P=0.003) and PDW≤15.90 (PFS: P=0.028, OS: P=0.006) were the independent risk factors for the prognosis of childhood medulloblastoma. Conclusions:Increased NLR and decreased PDW indicate unfavorable prognosis of the childhood medulloblastoma.Therefore, preoperative NLR and PDW may be the potential prognostic markers for childhood medulloblastoma.

PURPOSE: This study was aimed to investigate the effect of pseudolaric acid B (PAB) on proliferation, invasion and epithelial-to-mesenchymal transition (EMT) in pancreatic cancer cells and to explore the possible mechanism. MATERIALS AND METHODS: The pancreatic cancer cell line SW1990 was cultured and treated with PAB dose- and time-dependent manners. Cell proliferation and invasion ability were measured by MTT assay and Matrigel/Transwell test, respectively. Semi-quantitative real-time polymerase chain reaction and Western blotting were conducted to detect the expression of EMT markers and the key molecules. Finally, nude mice subcutaneous transplantation tumor model was used to confirm the therapy efficacy of PAB. RESULTS: PAB could inhibit SW1990 cell proliferation and invasion in time- and dose-dependent manners. Vimentin, fibronectin, N-cadherin, Snail, Slug, YAP, TEAD1, and Survivin were down-regulated (p < 0.01), while E-cadherin, caspase-9, MST1, and pYAP were up-regulated (p < 0.05). Combined PAB and gemcitabine treatment markedly restricted the tumor growth compared with gencitabin or PAB alone groups. CONCLUSION: PAB could inhibit the proliferation and invasion ability of pancreatic cancer cells through activating Hippo-YAP pathway and inhibiting the process of EMT.
Animals ; Antineoplastic Agents/pharmacology ; Antineoplastic Agents/therapeutic use ; Biomarkers, Tumor/metabolism ; Cadherins ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation/drug effects ; Cytokines ; Deoxycytidine/analogs & derivatives ; Deoxycytidine/pharmacology ; Deoxycytidine/therapeutic use ; Diterpenes/pharmacology ; Diterpenes/therapeutic use ; Epithelial-Mesenchymal Transition/drug effects ; Female ; Humans ; Mice, Nude ; Neoplasm Invasiveness ; Pancreatic Neoplasms/diet therapy ; Pancreatic Neoplasms/pathology ; Real-Time Polymerase Chain Reaction ; Signal Transduction/drug effects ; Vimentin/metabolism

Objective To explore the differences between ABI 7500 and TL 988 fluorescent quantitative instrument for polymerase chain reaction. Methods Based on EP9-A2 Files, quantitative influenza A virus positive controls of five kinds of concentrations were detected by two kinds of fluorescence quantitative PCR instruments at the same time, and each sample was detected 20 times. The precision and sensitivity between the two kinds of instruments were compared. Results Inner group CV of TL 988 ranged from 0.68% to 1.52%. The CV for ABI 7500 ranged from 1.93% to 2.69%. The high (107, 106, 105 copies/mL) levels of IFVA positive control were detected by ABI 7500 and TL 988 instruments, and there were no significant difference between the results(P>0.05) while the low(104, 103 copies /mL)levels were de-tected by ABI 7500 and TL 988 instruments, and there was a significant difference between the results(P<0.05). When samples were detected at the level of 103 copies/mL, the positive detection rate was 100% with TL 988 instrument, but the positive detection rate was 0 with ABI 7500 instrument. The related index of ABI 7500 and TL 988 was better (r=0.998). Conclusion The performances in precision and sensitivity of TL 988 PCR instrument are better than ABI 7500 PCR.

By analyzing the Military Health System winning "Heliang & Heli" Fund Award between 1994 and 2009,we found that the Military Health System possessed a good scientific basis and high level,but with an unbalanced scientific research development,which suggests the Military Health System should further strengthen its independent innovation ability.