Cardiovascular diseases resulting from atherosclerosis have become a serious threat to human health. It is well-known that an ongoing inflammatory response is involved during atherosclerosis progression that ultimately results in the accumulation of lipids and formation of plaques. Monitoring the pathological changes during the inflammatory response will be of great significance for early diagnosis and therapeutic evaluation of atherosclerosis. Targeted contrast-enhanced ultrasonography has been shown to be a promising noninvasive imaging technique for evaluating the degree of atherosclerosis and may potentially be translated to clinical imaging in the future. However, inadequate cell adhesion of targeted microbubbles (MBs) in large arterial vessels still remains a great challenge.Methods: By mimicking the leucocytes that are recruited to the vessel wall during the initiation of atherosclerosis through selectin-dependent arrest and cell adhesion molecule-mediated firm cell adhesion, we developed VCAM-1/ICAM-1/P-selectin-targeted MBVIS by integrating VCAM-1 and ICAM-1 antibodies and synthetic polymeric sialyl Lewis X (sLex) onto the MB surface.Results: The resulting MBVIS had a high affinity to inflammatory bEnd.3 cells in both static and dynamic flow conditions. Significantly enhanced ultrasound imaging signals were achieved by MBVIS in detecting the atherosclerosis progress when compared with the single- or dual-targeted MBs. Taking advantage of the artificial MBVIS, less ultrasound imaging signals were found in the atorvastatin-treated, but not placebo-treated, ApoE-deficient mice with atherosclerosis, revealing a potential therapeutic efficacy of atorvastatin for early stage atherosclerosis. This was further confirmed by histologic staining examination.Conclusions: Our study provides a promising ultrasound molecular imaging probe for early-stage diagnosis and therapeutic evaluation of atherosclerosis.

Background Placenta-derived MSCs (P-MSCs) represent a promising tool for cell-based therapeutic applications. However, the increasing demand for P-MSCs in clinical trials makes high quality and large number of P-MSCs mandatory. Here, we aim to develop an efficient protocol for P-MSC isolation and culture. Methods The modified explant culture (MEC) method by combining an initial mild enzymatic reaction with the subsequent explant culture was developed to simultaneously produce various P-MSCs from the different regions of the placenta in serum-free medium (SFM). Its isolation efficiencies, cell yield, and proliferative capacity were compared with the conventional explant culture (EC) method. Furthermore, we determined whether functional properties of P-MSCs are affected by the used tissue-harvesting sites in terms of their proliferation, migration, and the immunomodulatory effect on macrophage. Results The MEC method achieved higher yield and shorter time in primary cell confluence in SFM compared with the conventional method. The harvested cells possessed the MSC characteristics and demonstrated significantly stronger proliferation ability. Importantly, MSCs derived from chorionic plate (CP-MSCs) were found to exhibit superior properties to the other P-MSCs in proliferation and migration capacity, maintaining the fetal origin over serial passages. Notably, CP-MSCs show stronger ability in regulating macrophage polarization from M1 to M2. Conclusion Our study developed an efficient and high-yield technique to produce high-quality P-MSCs from the placenta, hence serving as an optimal source of MSCs for clinical application.

Background Liver cancer is one of the most common cancers worldwide. We aimed to report the burden of liver cancer at the global, regional, and national levels in 204 countries from 1990 to 2019, stratified by etiology, sex, age, and sociodemographic index (SDI). Methods Data of mortality, incidence, and disability‐adjusted life years (DALYs) of liver cancer and its etiology were available from the Global Burden of Diseases, Injuries, and Risk Factors (GBD) Study 2019. The trends in the liver cancer burden were assessed by the annual percentage change. All estimates are presented as numbers and age‐standardized rates (ASRs) per 100,000 population, with uncertainty intervals (UIs). Results Globally, 484,577 (95% UI 444,091–525,798) mortalities, 534,364 (486,550–588,639) incident cases, and 12,528,422 (11,400,671–13,687,675) disability‐adjusted life years (DALYs) due to liver cancer occurred in 2019. The ASRs were 5.95 (5.44–6.44), 6.51 (5.95–7.16), and 151.08 (137.53–164.8) per 100,000 population for the mortalities, incidences, and DALYs, respectively. From 1990 to 2019, the numbers increased, whereas the ASRs decreased. Hepatitis B and Hepatitis C are the major causes of liver cancer mortality. The liver cancer mortality in 2019 increased with age, peaking at 65–69 and 70–74 age group in males and females, respectively, and the number was higher in males than in females. Generally, there were nonlinear associations between the ASR and SDIs values at the regional and national levels. China had the highest numbers of mortalities, incident cases, and DALYs, whereas Mongolia has the highest ASR in 2019. Conclusion Liver cancer remains a major public health issue worldwide, but etiological and geographical variations exist. It is necessary to increase awareness of the population regarding liver cancer, its etiologies and the importance of early detection, and diagnosis and treatment.

Synthetic biology employs engineering principles to redesign biological systems for biomedical or industrial purposes. Innovation and application of original biological parts for genetic circuit construction will significantly facilitate and expedite the development of synthetic biology. Here, we built two‐ or three‐input linear double‐stranded DNA (ldsDNA)‐based Boolean AND gate genetic circuits in mammalian cells. Bioluminescence imaging revealed the feasibility of ldsDNA‐based Boolean AND gate circuits in vivo. Inhibition of DNA‐PKcs, a pivotal enzyme in nonhomologous end joining, significantly attenuated the output signals from ldsDNA‐based Boolean AND gate circuits. We further showed that ldsDNA with additional terminal random nucleotide(s) could undergo end nucleotide deletion and generate in‐frame proteins via the Boolean AND gate response. Additionally, ldsDNAs or plasmids with identical overlapping sequences could also serve as input signals for Boolean AND gate genetic circuits. Our work establishes ldsDNAs as innovative biological parts for building low noise‐signal ratio Boolean AND gate circuits with application potential in biomedical engineering fields.

A nanobody is an antibody fragment consisting of a single monomeric variable antigen-binding domain. Mammalian cells are ideal platforms for identifying nanobodies targeting hardto-display transmembrane proteins and nanobodies that function as modulators of cellular phenotypes. However, the introduction of a high-diversity nanobody library into mammalian cells is challenging. We have developed two novel methods for constructing a nanobody library in mammalian cells. Complementarity-determining region (CDR) random sequences were first incorporated into upstream and downstream dsDNAs by PCR. In the first method, named dsDNA-HR, upstream and downstream dsDNAs containing an identical overlapping sequence were co-transfected into cultured mammalian cells for intracellular homologous recombination that resulted in the formation of an intact nanobody library expression cassette. In the second method, named in vitro ligation, we generated fulllength nanobody expression dsDNAs via ligation of restriction digested upstream and downstream dsDNAs. The obtained fulllength dsDNAs were transfected into mammalian cells for nanobody library expression. Using both methods, we generated over a million unique nanobody sequences, as revealed by high-throughput sequencing. Single-cell sequencing was employed to resolve the diversity of the dsDNA-HR nanobody library. We also identified a small molecule, Nocodazole, which could enhance the efficacy of dsDNA-HR.