Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) are widely used as gene editing tools in biology, microbiology, and other fields. CRISPR is composed of highly conserved repetitive sequences and spacer sequences in tandem. The spacer sequence has homology with foreign nucleic acids such as viruses and plasmids; Cas effector proteins have endonucleases, and become a hotspot in the field of molecular diagnosis because they recognize and cut specific DNA or RNA sequences. Researchers have developed many diagnostic platforms with high sensitivity, high specificity, and low cost by using Cas proteins (Cas9, Cas12, Cas13, Cas14, etc.) in combination with signal amplification and transformation technologies (fluorescence method, lateral flow technology, etc.), providing a new way for rapid detection of pathogen nucleic acid. This paper introduces the biological mechanism and classification of CRISPR-Cas technology, summarizes the existing rapid detection technology for pathogen nucleic acid based on the trans cleavage activity of Cas, describes its characteristics, functions, and application scenarios, and prospects the future application of this technology.

In the field of liver disease treatment, liver transplantation (LT) has become an effective option for end-stage liver disease. However, issues such as immune rejection and graft damage remain important factors influencing the success rate of liver transplantation and patients' quality of life. In recent years, with the advancement of genetic testing technologies, the study and application of donor-derived cell-free DNA (dd-cfDNA) in LT diagnosis and treatment have gradually gained attention. This review explores the research advancements in dd-cfDNA within liver transplant management, evaluating its potential applications throughout the liver transplantation process, while exploring the challenges faced by current studies and outlining future research directions. As a strategic tool for postoperative monitoring in LT, dd-cfDNA shows promising potential in areas such as immune rejection, graft damage, immunosuppressant adjustment, complication monitoring, and personalized treatment, and is poised to become a reliable biomarker in LT management.

Hypothermic oxygenated machine perfusion (HOPE) can enhance organ preservation and protect mitochondria from hypoxia-ischemic injury; however, an understanding of the underlying HOPE mechanism that protects mitochondria is somewhat lacking. We hypothesized that mitophagy may play an important role in HOPE mitochondria protection. Experimental rat liver grafts were exposed to 30 min of in situ warm ischemia. Then, grafts were procured, followed by cold storage for 3 or 4 h to mimic the conventional preservation and transportation time in donation after circulatory death (DCD) in clinical contexts. Next, the grafts underwent hypothermic machine perfusion (HMP) or HOPE for 1 h through portal vein only perfusion. The HOPE-treated group showed a better preservation capacity compared with cold storage and HMP, preventing hepatocyte damage, nuclear injury, and cell death. HOPE can increase mitophagy marker expression, promote mitophagy flux via the PINK1/Parkin pathway to maintain mitochondrial function, and reduce oxygen free radical generation, while the inhibition of autophagy by 3-methyladenine and chloroquine could reverse the protective effect. HOPE-treated DCD liver also demonstrated more changes in the expression of genes responsible for bile metabolism, mitochondrial dynamics, cell survival, and oxidative stress. Overall, HOPE attenuates hypoxia-ischemic injury in DCD liver by promoting mitophagy flux to maintain mitochondrial function and protect hepatocytes. Mitophagy could pave the way for a protective approach against hypoxia-ischemic injury in DCD liver.

Objective: For heart transplantation, donor heart status needs to be evaluated during normothermic ex situ perfusion (ESHP). Left ventricular end-systolic elastance (Ees) measures the left ventricular contractile function, but its estimation requires the occlusion of the left atrium line in the ESHP, which may cause unnecessary damage to the donor heart. We present a novel method to quantify Ees based on hemodynamic parameters obtained from only one steady-state PV loop in ESHP. Methods: Ees was obtained by the end-systolic point (Pes, Ves) and the volume axis intercept point of Ees (V0). V0 was estimated through the support vector machine regression (SVR) method using parameters derived from the measured steady-state PV loop. To achieve high V0 estimation accuracy, a filter-based support vector machine recursive feature elimination method (SVM-RFE) algorithm selected the parameters for V0 estimation. Hemodynamic parameter samples (n = 101) obtained from ESHP experiments with pig s hearts were used to train the Ees calculation model. Early post-transplantation outcomes in six heart transplantation experiments were then estimated from the trained Ees calculation model. Results: Ees calculated by the proposed method agreed well with conventional multi-beat estimates obtained by the occlusion process (r = 0.88, p < 0.001, n = 101) and was capable of predicting the early post-transplant cardiac index (r = 0.84, p < 0.05, n = 6). Conclusion: This method effectively assesses left ventricular contractility during ESHP and predicts early post-transplant outcomes in the porcine model. Significance: Our approach is the first to quantify Ees by estimating V0 from steady-state beats in ESHP for accurately predicting early post-transplantation outcomes.

Background: Investigating basal blood perfusion in the major arteries of the limbs to guide flow rate selection during ex vivo perfusion preservation for limb replantation (transplantation).Methods: Volunteers undergoing physical examinations at PLAGH Fourth Medical Center (February–December 2024) were recruited. Three hundred and four eligible participants (146 males, 158 females; aged 18–65, mean 41.68 ± 11.28) were enrolled after screening. A portable Doppler ultrasound system was utilized to assess the brachial, ulnar, radial, popliteal, dorsalis pedis, and posterior tibial arteries in the limbs. Vascular diameter, blood flow velocity, and blood perfusion volume were measured for each artery. Mean hemodynamic parameters were calculated. Analyze the linear relationship between BMI, body surface area, and arterial blood perfusion volume using a multiple linear regression model. Conduct interaction tests to investigate whether there are sex-specific or “three-high” status-specific effects in the linear prediction model of blood perfusion volume by BMI and body surface area.Results: Ultrasound findings revealed the following mean blood perfusion volumes in the upper limb arteries: brachial artery, 74.9 ± 22.5 mL/min; ulnar artery, 35.7 ± 12.6 mL/min; radial artery, 36.8 ± 13.5 mL/min. In the lower limb arteries, the mean blood perfusion volume was: popliteal artery, 114.1 ± 34.2 mL/min; dorsalis pedis artery, 53.3 ± 18.1 mL/min; posterior tibial artery, 59.2 ± 21.0 mL/min. No significant difference was observed in mean blood perfusion volume between the ulnar and radial arteries (p > 0.05), whereas the posterior tibial artery exhibited significantly higher blood perfusion volume compared to the dorsalis pedis artery (p < 0.001). Multiple linear regression analysis revealed that BMI was negatively associated with arterial blood perfusion volume in the extremities, while body surface area showed a positive association. Furthermore, BMI and body surface area jointly formed a linear predictive relationship with limb blood flow. Based on significant effects within the linear model and pathophysiological mechanisms, interaction terms for body surface area (BSA) × BMI and age × diabetes status were included. The results demonstrated a statistically significant interaction effect (p < 0.05) between BMI and body surface area on limb blood flow. However, the interaction effect of diabetes status on limb blood flow was not significant (p > 0.05).Conclusion: Quantitative ultrasound-derived limb perfusion parameters and their BMI/BSA correlations enable hemodynamic customization for machine perfusion systems in limb replantation. This standard approach balances metabolic support and ischemia-reperfusion risk mitigation during extracorporeal preservation, advancing personalized transplant protocols.

Cytomegalovirus (CMV) infection is a highly prevalent opportunistic infection among liver transplant recipients. When the liver donor is infected with CMV, there is a risk of transmission to the recipient, leading to CMV infection. To improve the postoperative outcome of liver transplantation, it is crucial to shift the focus of CMV detection to the donor and achieve early diagnosis, as well as implement effective preventative and therapeutic measures. However, the commonly used CMV detection methods in the past had limitations that prevented their early and accurate diagnosis in liver transplant donors. This review focuses on the latest advancements in CMV detection methods that can potentially be applied to liver transplant donors. The objective is to compare and evaluate their clinical utility, thereby providing guidance and support for rapid and accurate diagnosis of CMV infection in the clinic. The clustered regularly interspaced short palindromic repeats-associated proteins (CRISPR–Cas) system-based assay emerges as a promising method for detecting the virus, offering great prospects for early and expedient CMV infection diagnosis in clinical settings.