Ischemic stroke is a catastrophic event in young adults. It may cause death,disability, incapacity and decline in the quality of life. When the neurologists are facing the suspected ischemic stroke in young adults, the greatest challenge of diagnosis is to identify the causes. This article reviews the risk factors, etiological diagnosis as well as its associated disorders and dysfunction in young adults.

ObjectiveTo investigate whether the reference intervals for serum alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) in WS/T 645.2-2018 are applicable to the adults in Changchun, China. MethodsAccording to the validation method for the reference intervals in WS/T 645.2-2018, 5420 subjects, aged from 20-79 years, who underwent physical examination from January 2016 to April 2019 were enrolled and divided into 12 subgroups based on age and sex. Each subgroup was analyzed in terms of whether more than 90% of the measured values fell into the reference intervals, and the changing trend of AFP and CEA with age and sex was analyzed. The Mann-Whitney U test was used for comparison between two groups. ResultsMore than 90% of the measured values of serum AFP and CEA fell into the reference intervals, which passed the validation test. There was a significant difference in serum AFP between male and female subjects aged 30-39 years (Z=-4.51, P＜0.05), and there was a significant difference in serum CEA between male and female subjects aged 20-29, 30-39, 40-49, 50-59, and 60-69 years (Z=-13.45, -18.15, -17.34, -10.82, and -3.65, all P＜0.05). Serum AFP increased slowly with age in female subjects aged 20-69 years and decreased with age in male and female subjects aged 70-79 years. Serum CEA increased with age in male and female subjects aged 20-79 years, and male subjects had a higher measured value than female subjects. ConclusionReference intervals for serum AFP and CEA in WS/T 645.2-2018 issued by National Health Commission are applicable to the adults in Changchun.

Background/Aims: This study aimed to investigate the biological function and regulatory mechanism of TCN1 in colorectal cancer (CRC). Methods: We studied the biological function of TCN1 by performing gain-of-function and loss-offunction analyses in HCT116 cell lines; examined the effects of TCN1 on the proliferation, apoptosis, and invasion of CRC cells; and determined potential molecular mechanisms using HCT116 and SW480 CRC lines and mouse xenotransplantation models. Tumor xenograft and colonization assays were performed to detect the tumorigenicity and metastatic foci of cells in vivo. Results: TCN1 knockdown attenuated CRC cell proliferation and invasion and promoted cell apoptosis. Overexpression of TCN1 yielded the opposite effects. In addition, TCN1-knockdown HCT116 cells failed to form metastatic foci in the peritoneum after intravenous injection. Molecular mechanism analyses showed that TCN1 interacted with integrin subunit β4 (ITGB4) to positively regulate the expression of ITGB4. TCN1 knockdown promoted the degradation of ITGB4 and increased the instability of ITGB4 and filamin A. Downregulation of ITGB4 at the protein level resulted in the disassociation of the ITGB4/plectin complex, leading to cytoskeletal damage. Conclusions TCN1 might play an oncogenic role in CRC by regulating the ITGB4 signaling pathway.

Background/Aims: Colorectal cancer (CRC) patients often exhibit peritoneal metastasis, which negatively impacts their prognosis. CD31 and D2-40 have recently been suggested to be predictors of breast cancer prognosis, but their role in colorectal peritoneal metastasis (CRPM) remains unknown. Methods: The expression profiles of CD31 and D2-40 were analyzed in CRC patients with or without CRPM and in CRC cell lines with increasing metastatic potential. Overexpression and short hairpin RNA knockdown assays were performed in CRC cells, and the effects of these alterations on epithelial-mesenchymal transition (EMT) in vitro, growth of xenograft tumors in vivo, and peritoneal metastasis potential in a mouse model of CRPM were examined. Results: The expressions of CD31 and D2-40 were upregulated in CRC tumor tissues and was elevated further in tumor tissues from patients with CRPM. CD31 and D2-40 expression levels exhibited increasing trends parallel to the EMT potential of CRC cells. CD31 and D2-40 are essential for CRC cell EMT in vitro as well as for xenograft tumor growth and peritoneal metastasis in vivo. Conclusions CD31 and D2-40 contribute to CRPM by promoting EMT and may serve as prognostic markers and therapeutic targets for CRC, particularly in patients with peritoneal metastasis.

Objective: To establish reference intervals (RIs) of thyroid hormone based on data from healthy subjects in laboratory information system (LIS) by indirect methods. Methods: Data were selected from the physical examination center in LIS of the First Hospital of Jilin University from May 2014 to December 2018. The normal distribution of the original data was checked by the Kolmogorov-Smirnov test. Skewed data were transformed into normal distribution using BOX-COX techniques, and outliers were identified by the Turkey method. The continuous percentile curve was established by coefficient of skewness-median-coefficient of variation(LMS) methods. Cut-off value of age was determined by decision trees, and the differences between groups were verified by Z-tests. P_2.5 and P_97.5 in the RIs were analyzed by non-parametric methods. Results: A total of 45 742 subjects were included in the study. There were no differences in the RI for thyroid stimulating hormone (TSH) among age groups or between men and women (Z*), and the RI was 0.60-4.41mIU/L (20-79 years old). However, the levels of free triiodothyronine(FT₃) decreased with age in males and females under 35-year-old. The RIs for FT₃ were 4.47-6.44pmol/L (20-44 years old), 4.19-6.21pmol/L (45-64 years old) and 3.90-5.85pmol/L (65-79 years old) in males, and 4.04-6.13pmol/L (20-34 years old) and 3.87-5.76pmol/L (35-79 years old) in females, respectively. The levels of free thyroxine(FT₄) in men were higher than those in women from 20 to 49 years old, and no gender and age differences could be viewed in subjects after 50 years old (Z*). The RIs for FT₄ were 13.69-21.76pmol/L (male, 20-49 years old), 12.99-20.83pmol/L (female, 20-49 years old) and 12.98-21.21pmol/L (50~79 years old). Conclusion Establishment of RIs of thyroid hormone based on the data from LIS is simple and reliable, which is suitable for clinical laboratory application.

Gene expression labeling and conditional manipulation of gene function are important for elaborate dissection of gene function. However, contemporary generation of pairwise dual-function knockin alleles to achieve both conditional and geno-tagging effects with a single donor has not been reported. Here we first developed a strategy based on a flipping donor named FoRe to generate conditional knockout alleles coupled with fluorescent allele-labeling through NHEJ-mediated unidirectional targeted insertion in zebrafish facilitated by the CRISPR/Cas system. We demonstrated the feasibility of this strategy at sox10 and isl1 loci, and successfully achieved Cre-induced conditional knockout of target gene function and simultaneous switch of the fluorescent reporter, allowing generation of genetic mosaics for lineage tracing. We then improved the donor design enabling efficient one-step bidirectional knockin to generate paired positive and negative conditional alleles, both tagged with two different fluorescent reporters. By introducing Cre recombinase, these alleles could be used to achieve both conditional knockout and conditional gene restoration in parallel; furthermore, differential fluorescent labeling of the positive and negative alleles enables simple, early and efficient real-time discrimination of individual live embryos bearing different genotypes prior to the emergence of morphologically visible phenotypes. We named our improved donor as Bi-FoRe and demonstrated its feasibility at the sox10 locus. Furthermore, we eliminated the undesirable bacterial backbone in the donor using minicircle DNA technology. Our system could easily be expanded for other applications or to other organisms, and coupling fluorescent labeling of gene expression and conditional manipulation of gene function will provide unique opportunities to fully reveal the power of emerging single-cell sequencing technologies.
Alleles ; Animals ; CRISPR-Cas Systems ; DNA End-Joining Repair ; DNA, Circular/metabolism* ; Embryo, Nonmammalian ; Gene Editing/methods* ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Genes, Reporter ; Genetic Loci ; Genotyping Techniques ; Green Fluorescent Proteins/metabolism* ; Integrases/metabolism* ; Luminescent Proteins/metabolism* ; Mutagenesis, Insertional ; Single-Cell Analysis ; Zebrafish/metabolism*