The Protective Effects of Trypsin Inhibitor on Hepatic Ischemia-Reperfusion Injury and Liver Graft Survival

The aim of this study was to explore the protective effects of ulinastatin (urinary trypsin inhibitor, UTI) on liver ischemia-reperfusion injury (IRI) and graft survival. We employed mouse liver cold IRI and orthotopic liver transplantation (OLTx) models. UTI was added to lactated Ringer’s (LR) solution for liver perfusion and preservation in vitro or combined with UTI injection intraperitoneally to the liver graft recipient. Our results indicated that UTI supplementation protected the liver from cold IRI in a dose-dependent manner and prolonged liver graft survival from extended cold preserved liver donors significantly. The underlying mechanism of UTI on liver IRI may be mediated by inhibition of proinflammatory cytokine release, increasing the expression of the antiapoptotic gene Bcl-2 and decreasing the expression of the proapoptosis genes of Caspase-3 and Bax, and further protects hepatocytes from apoptotic death and improves liver function

Expression of DNMT1 and DNMT3a Are Regulated by GLI1 in Human Pancreatic Cancer

BACKGROUND AND AIMS: GLI1, as an indispensable transcriptional factor of Hedgehog signaling pathway, plays an important role in the development of pancreatic cancer (PC). DNA methyltransferases (DNMTs) mediate the methylation of quantity of tumor-related genes. Our study aimed to explore the relationship between GLI1 and DNMTs. METHODS: Expressions of GLI1 and DNMTs were detected in tumor and adjacent normal tissues of PC patients by immunohistochemistry (IHC). PANC-1 cells were treated by cyclopamine and GLI1-siRNA, while BxPC-3 cells were transfected with overexpression-GLI1 lentiviral vector. Then GLI1 and DNMTs expression were analyzed by qRT-PCR and western blot (WB). Then we took chromatin immunoprecipitation (ChIP) to demonstrate GLI1 bind to DNMT1. Finally, nested MSP was taken to valuate the methylation levels of APC and hMLH1, when GLI1 expression altered. RESULTS: IHC result suggested the expressions of GLI1, DNMT1 and DNMT3a in PC tissues were all higher than those in adjacent normal tissues (p<0.05). After GLI1 expression repressed by cyclopamine in mRNA and protein level (down-regulation 88.1±2.2%, 86.4±2.2%, respectively), DNMT1 and DNMT3a mRNA and protein level decreased by 91.6%±2.2% and 83.8±4.8%, 87.4±2.7% and 84.4±1.3%, respectively. When further knocked down the expression of GLI1 by siRNA (mRNA decreased by 88.6±2.1%, protein decreased by 63.5±4.5%), DNMT1 and DNMT3a mRNA decreased by 80.9±2.3% and 78.6±3.8% and protein decreased by 64.8±2.8% and 67.5±5.6%, respectively. Over-expression of GLI1 by GLI1 gene transfection (mRNA increased by 655.5±85.9%, and protein increased by 272.3±14.4%.), DNMT1 and DNMT3a mRNA and protein increased by 293.0±14.8% and 578.3±58.5%, 143.5±17.4% and 214.0±18.9%, respectively. ChIP assays showed GLI1 protein bound to DNMT1 but not to DNMT3a. Results of nested MSP demonstrated GLI1 expression affected the DNA methylation level of APC but not hMLH1 in PC. CONCLUSION: DNMT1 and DNMT3a are regulated by GLI1 in PC, and DNMT1 is its direct target gene

CRISPR/Cas9‐mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse

The X‐linked genetic bleeding disorder caused by deficiency of coagulator factor IX, hemophilia B, is a disease ideally suited for gene therapy with genome editing technology. Here, we identify a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. The CRISPR/Cas9 system was used to generate distinct genetically modified mouse models and confirmed that the novel Y371D mutation resulted in a more severe hemophilia B phenotype than the previously identified Y371S mutation. To develop therapeutic strategies targeting this mutation, we subsequently compared naked DNA constructs versus adenoviral vectors to deliver Cas9 components targeting the F9 Y371D mutation in adult mice. After treatment, hemophilia B mice receiving naked DNA constructs exhibited correction of over 0.56% of F9 alleles in hepatocytes, which was sufficient to restore hemostasis. In contrast, the adenoviral delivery system resulted in a higher corrective efficiency but no therapeutic effects due to severe hepatic toxicity. Our studies suggest that CRISPR/Cas‐mediated in situ genome editing could be a feasible therapeutic strategy for human hereditary diseases, although an efficient and clinically relevant delivery system is required for further clinical studies

Large genomic fragment deletion and functional gene cassette knock-in via Cas9 protein mediated genome editing in one-cell rodent embryos

The CRISPR-Cas RNA-guided system has versatile uses in many organisms and allows modification of multiple target sites simultaneously. Generating novel genetically modified mouse and rat models is one valuable application of this system. Through the injection of Cas9 protein instead of mRNA into embryos, we observed fewer off-target effects of Cas9 and increased point mutation knock-in efficiency. Large genomic DNA fragment (up to 95 kb) deletion mice were generated for in vivo study of lncRNAs and gene clusters. Site-specific insertion of a 2.7 kb CreERT2 cassette into the mouse Nfatc1 locus allowed labeling and tracing of hair follicle stem cells. In addition, we combined the Cre-Loxp system with a gene-trap strategy to insert a GFP reporter in the reverse orientation into the rat Lgr5 locus, which was later inverted by Cre-mediated recombination, yielding a conditional knockout/reporter strategy suitable for mosaic mutation analysis

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Building a Cultural Bridge: A study of intercultural communication between Sweden and China

With the increasing interactions between Sweden and China, it is interesting to see how people of these two countries handle their intercultural communication. Based on my internship experiences, where I encountered many cases of intercultural communication between Sweden and China, this thesis presents some contributions to a better understanding of Swedish‐Sino communications. The following questions are discussed: • Where do communication misunderstandings come into play? • How do individuals get around cultural misunderstandings? • What factors facilitate understandings in intercultural contexts between Swedes and Chinese? Through the analyses of three difference cases, the significance of this thesis shows that: • Culture is a process. • Cultural differences exist not only between cultures, but also within cultures. • Intercultural encounters do not always end up as cultural misunderstandings, while much of the literature on intercultural communication dwells on the issue of difference and misunderstanding. This thesis argues that intercultural communication needs to nurture a more reflective ethic of difference in which difference is not only recognized and tolerated, but in which recognition and tolerance should be seen as a precondition for the development of new forms of understanding, in the face of difference. At the end of this thesis, the factors that facilitate understanding in intercultural relations between Swedes and Chinese are discussed. It also emphasizes the important role an intermediary plays in intercultural communication and the reader should gain better, more accurate and in depth understanding of Swedish‐Sino communication. <BR

Characteristics and Development Mechanisms of Northeast Cold Vortices

The northeast cold vortices (NECVs) in May-September during 1989–2018 are classified, based on the 6 h NCEP/NCAR reanalysis data (2.5° × 2.5°) and observational data from the Meteorological Information Comprehensive Analysis and Process System (MICAPS) provided by China Meteorological Administration. Meanwhile, characteristics and development mechanisms for NECVs of different types are also analyzed. In the recent 30 years, the occurrences of NECV processes have been increasing year by year, with an average of 7.4 times per year in Northeast China and a duration of 3–5 days on average for each process. NECVs mostly occur in late spring and early summer, and the longest time influenced by NECVs exceeds 19 days, with annual means of 9.9 days, 8.8 days, and 7.0 days in May, June, and July, respectively. The frequency of weak NECVs is about 1.2 times that of strong NECVs. Strong NCVs in late spring and early autumn as well as weak MCVs in summer are with high-frequency occurrences. It is found that when NCVs occur in late spring and early autumn, the upper-level westerly jets are relatively stronger, thus strengthening the divergence in the upper troposphere and the vortex circulation. The circulation fields in upper and lower levels cooperate with the strong jets, promoting the continuous development and maintenance of the cold vortices. Apart from the jets and circulation, the lower central potential height combined with the obvious cold-core and stronger ascending motions favor the NCV’s development. In addition, the dry intrusion has a strong promotion due to the stronger lower-level cold advection and downward intrusion of high potential vorticity. However, when MCVs occur in summer, things are just the opposite