Objective:To investigate the long-term prognosis of liver transplantation (LT) recipients who received grafts from donors aged ≥80 years and the associated risk factors.Methods:Clinical and follow-up data of LT recipients from January 2002 to June 2023 were retrospectively analyzed using the United Network for Organ Sharing (UNOS) database. Donors were categorized into three groups : non-elderly donors (NED, age < 60 years), elderly donors (ED, age 60-79 years), and very elderly donors (VED, age ≥80 years). Propensity score matching (PSM) was used to reduce baseline selection bias among the groups. Survival rates were calculated using the Kaplan-Meier method, and differences among the groups were compared using the Log-Rank test. Univariate and multivariate Cox proportional hazards regression analyses were performed to identify independent risk factors for overall survival (OS) in the VED group. Recipients were further subdivided into three age groups (<50 years, 50-69 years, and ≥70 years) to compare survival outcomes among NED, ED, and VED groups.Results:A total of 115, 089 LT recipients were included, comprising 95, 973 (83.4%) in the NED group, 18, 520 (16.1 %) in the ED group, and 596 (0.5 %) in the VED group. After 3∶3∶1 PSM, each group included 1, 623 recipients for NED and ED, and 541 for VED, with no significant differences in baseline data. The 10-year OS rates for NED, ED, and VED groups were 61.8%, 55.9%, and 47.8%, respectively, and the 10-year graft survival (GS) rates were 61.3 %, 53.8%, and 45.9 %, respectively, with all comparisons showing statistical significance ( P< 0.001). In recipients aged <70 years, the VED group had significantly lower OS and GS rates (49.0% and 47.1 %, respectively ) compared to the NED (63.7 % and 61.8%) and ED (57.7% and 55.2 %) groups ( P< 0.001 ). For recipients aged ≥70 years, there were no significant differences in 10-year OS and GS among the NED (47.2% and 48.7 %), ED (47.0 % and 48.7 %), and VED (40.0 % and 39.2 %) groups ( P= 0.992 and P= 0.996, respectively). Cox regression analysis identified cold ischemia time ≥8 hours ( HR=1.447, 95% CI: 1.088-1.923, P=0.011), pre-transplant ICU dependence ( HR=1.803, 95% CI: 1.176–2.765, P=0.007), and hepatitis B/C virus infection ( HR =1.432, 95% CI: 1.057-1.941, P=0.020) as independent risk factors for OS in the VED group. Conclusions:Liver grafts from VED grafts significantly reduce long-term OS and GS in recipients, except in those aged ≥70 years where prognosis is comparable to recipients of NED and ED grafts.. For the VED group, factors such as cold ischemia time ≥8 hours, pre-transplant ICU dependence, and hepatitis B/C virus infection markedly influence the prognosis.

OBJECTIVETo investigate EXT1 and EXT2 genes mutations in a family with hereditary multiple osteochondromas (HME).

METHODSA four-generation family with HME from Linyi city of Shandong Province was studied. There were 6 affected individuals among the 17 family members. Physical examination and radiographical evaluations were carried out for all family members. Genomic DNA was extracted from peripheral venous blood and the samples were subjected to mutation screening by PCR of the coding regions of EXT1 and EXT2 genes.

RESULTSThe family has featured an autosomal dominant inheritance pattern. Sequencing of the EXT1 and EXT2 genes suggested the causative gene in this family was in linkage with the second exon of EXT2. A c.244delG mutation was detected, which has resulted in a frameshift mutation p.Asp81IlefsX30. The mutation was found in all of the 6 affected individuals but not in normal family members. And the mutation has co-segregated with the phenotype.

CONCLUSIONThe mutation c.244delG in the EXT2 gene is the probably the cause of the disease in this family.

Adult ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Pedigree ; Point Mutation ; Young Adult

The aim of this study was to develop a blocking enzyme-linked immunosorbent assay (bELISA) based on a biotinylated nanobody target the S1 protein of porcine epidemic diarrhea virus (PEDV) for detecting the anti-PEDV antibodies and evaluating the immune effect of the vaccine. The gene encoding the single-domain antibody sdAb3 target the PEDV S1 protein was amplified and the Avitag sequence was fused at its 3'-end. The PCR product was cloned into the expression vector pET-21b for expression and purification of the sdAb3-Avitag protein. The purified sdAb3-Avitag fusion protein was biotinylated and its activity was determined. Using the recombinant S1 protein as a coating antigen, a bELISA was established and optimized. Serum samples were tested in parallel by the bELISA and a commercial kit. The recombinant vector pET21b-sdAb3-Avitag was constructed to express the tagged sdAb3. After induction for expression, the biotin-labeled sdAb3 (sdAb3-Biotin) with high purity and good activity was obtained. For the optimized bELISA, the coating concentration of the S1 protein was 200 ng/well, the serum dilution was 1:2 and incubated for 2 h, the dilution ratio of the biotinylated sdAb3 was 1:8 000 and incubated for 30 min, the dilution of the enzyme-labeled antibody was 1:5 000 and incubated for 30 min. The bELISA had no cross reaction with the sera of major porcine viruses including transmissible gastroenteritis virus, porcine reproductive and respiratory syndrome virus and showed good specificity and reproducibility. For a total of 54 porcine serum samples tested, the overall compliance rate of the bELISA with a commercial kit was 92.56%. This study developed a rapid and reliable bELISA method, which can be used for serosurveillance and vaccine evaluation for PEDV.
Animals ; Antibodies, Viral ; Coronavirus Infections/veterinary* ; Enzyme-Linked Immunosorbent Assay ; Porcine epidemic diarrhea virus/genetics* ; Reproducibility of Results ; Sensitivity and Specificity ; Single-Domain Antibodies ; Swine ; Swine Diseases

The coronavirus disease 2019 (COVID-19) pandemic is caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is spread primary via respiratory droplets and infects the lungs. Currently widely used cell lines and animals are unable to accurately mimic human physiological conditions because of the abnormal status of cell lines (transformed or cancer cells) and species differences between animals and humans. Organoids are stem cell-derived self-organized three-dimensional culture in vitro and model the physiological conditions of natural organs. Here we showed that SARS-CoV-2 infected and extensively replicated in human embryonic stem cells (hESCs)-derived lung organoids, including airway and alveolar organoids which covered the complete infection and spread route for SARS-CoV-2 within lungs. The infected cells were ciliated, club, and alveolar type 2 (AT2) cells, which were sequentially located from the proximal to the distal airway and terminal alveoli, respectively. Additionally, RNA-seq revealed early cell response to virus infection including an unexpected downregulation of the metabolic processes, especially lipid metabolism, in addition to the well-known upregulation of immune response. Further, Remdesivir and a human neutralizing antibody potently inhibited SARS-CoV-2 replication in lung organoids. Therefore, human lung organoids can serve as a pathophysiological model to investigate the underlying mechanism of SARS-CoV-2 infection and to discover and test therapeutic drugs for COVID-19.
Adenosine Monophosphate/therapeutic use* ; Alanine/therapeutic use* ; Alveolar Epithelial Cells/virology* ; Antibodies, Neutralizing/therapeutic use* ; COVID-19/virology* ; Down-Regulation ; Drug Discovery ; Human Embryonic Stem Cells/metabolism* ; Humans ; Immunity ; Lipid Metabolism ; Lung/virology* ; RNA, Viral/metabolism* ; SARS-CoV-2/physiology* ; Virus Replication/drug effects*

Depression is a mental disorder with high morbidity, disability and relapse rates. Ginkgo biloba extract (GBE), a traditional Chinese medicine, has a long history of clinical application in the treatment of cerebral and mental disorders, but the key mechanism remains incompletely understood. Here we showed that GEB exerted anti-depressant effect in mice through regulating gut microbial metabolism. GBE protected against unpredictable mild stress (UMS)-induced despair, anxiety-like and social avoidance behavior in mice without sufficient brain distribution. Fecal microbiome transplantation transmitted, while antibiotic cocktail abrogated the protective effect of GBE. Spatiotemporal bacterial profiling and metabolomics assay revealed a potential involvement of Parasutterella excrementihominis and the bile acid metabolite ursodeoxycholic acid (UDCA) in the effect of GBE. UDCA administration induced depression-like behavior in mice. Together, these findings suggest that GBE acts on gut microbiome-modulated bile acid metabolism to alleviate stress-induced depression.
Humans ; Mice ; Animals ; Depression/drug therapy* ; Gastrointestinal Microbiome ; Plant Extracts ; Ginkgo biloba