[Complexity of Detecting CRISPR/Cas9-Mediated Homologous Recombination in Zebrafish].

Homology-directed (HD) genome modification offers an opportunity to precisely modify the genome. Despite reported successful cases, for many loci, precise genome editing remains challenging and inefficient in vivo. Here we report an effort to precisely knock-in a GFP reporter into gad locus mediated by CRISPR/Cas9 system in the zebrafish Danio rerio. PCR artifact was detected in testing for homologous recombination (HR), but was mitigated by optimizing PCR condition and decreasing the injected targeting plasmid concentration. Under this optimized condition, time course analysis revealed a decline of the HR-positive embryos at embryogenesis progressed. GFP signals also diminished at later developmental stages. The GFP signals were consistent with PCR detection, both of which suggested the loss of targeted insertion events at later stages. Such loss of insertion might be one underlying reason for the inability to obtain germ-line transgenic lines with GFP knocked into the gad locus. Our results suggest that the low HR efficiency associated with CRISPR-mediated knock-in is in part due to loss of insertion after targeted integration into the gad locus.

Specific genes related to nucleopolyhedrovirus in Bombyx mori susceptible and near-isogenic resistant strains through comparative transcriptome analysis.

Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the primary pathogens that causes severe economic losses to sericulture. Comparative transcriptomics analysis has been widely applied to explore the antiviral mechanism in resistant strains. Here, to identify genes involved in BmNPV infection, we identified differentially expressed genes (DEGs) and performed weighted gene co-expression network analysis (WGCNA) between two Bombyx mori strains: strain 871 (susceptible to BmNPV infection) and the near-isogenic strain 871C (resistant to BmNPV). Our results showed that 400 genes were associated with resistance in strain 871C, and 76 genes were related to susceptibility in strain 871. In addition, the correlation analysis of DEGs and WGCNA showed that 40 genes related to resistance were highly expressed in the resistant strain. Among them, gene BGIBMGA004291 was the most noticeable. We further identified the effect of gene BGIBMGA004291, which encoded a multiprotein bridge factor 2 (MBF2) family member (MBF2-10), on viral infection in cells. Our data suggested that MBF2-10 inhibited viral infection. Taken together, this study showed specific module trait correlations related to viral infection in strains 871 and 871C, and we identified a resistance-related gene. These findings suggested promising candidate genes with antiviral activity, aiding in the analysis of the antiviral molecular mechanisms in resistant strains.

miR-16-1-3p targets TWIST1 to inhibit cell proliferation and invasion in NSCLC.

In our study, the impact of miR-16-1-3p on cell proliferation and invasion in NSCLC was explored. miR-16-1-3p mimics were transfected to A549 cells for miR-16-1-3p overexpression. qRT- PCR and Western blot were applied to explore the relative expression of mRNA and protein in A549 cells. Furthermore, the cell proliferation capability was determined by MTT assay. Additionally, cell migration and invasion were measured using a scratch assay and transwell assay, respectively. Moreover, TargetScan and luciferase reporter assay was utilized to investigate the target of miR-16-1-3p. The results indicated that miR-16-1-3p was downregulated in NSCLC cells and upregulation of miR-16-1-3p was able to inhibit the expression of TWIST1. In addition, the reduced cell proliferation, inhibited cell migration and invasion were observed in miR-16-1-3p mimic group compared to the negative control group. The luciferase reporter gene showed that TWIST1 was a target of miR-16-1-3p. Therefore, the present study demonstrated that miR-16-1-3p may suppress A549 cell proliferation, migration and invasion by targeting TWIST1. Thus, miR-16-1-3p might play important roles in NSCLC development, which provides a novel aspect for NSCLC investigation (Fig. 6, Ref. 26).

[Research advances of collagen-based biomaterials in wound repair].

Acute or chronic wounds are common clinical problems. Collagen, with advantages including rich sources, impeccable biocompatibility, and inherent biodegradability, has been widely used in fundamental research and clinical treatment of wound repair with broad prospects of clinical applications. This article provided a brief overview of the role of collagen in various biological processes related to wound healing and also outlined the sources of collagen. Furthermore, the article summarized the application and recent research advancements of collagen-based wound dressings in the field of wound repair.

[Progress on the mechanism and application of adipose-derived stem cells in promoting wound repair].

Chronic refractory wounds and scars caused by abnormal wound repair seriously damage the health of patients and affect their quality of life. At present, there is a lack of simple but effective and economical treatment methods. Adipose-derived stem cells (ASCs), as a kind of mesenchymal stem cells with multi-directional differentiation potential, have been confirmed by several in vivo and in vitro studies to promote wound healing by promoting epithelialization, angiogenesis, immunoregulation, antioxidant properties, and other mechanisms. ASCs and their derivatives have been used in the treatment of refractory wounds caused by burns, diabetic, and radiation injuries with good results achieved. Their potential to become new materials for wound repair has also been confirmed. This paper reviewed the mechanism and clinical application of ASCs in promoting wound repair, and looked into its research direction and prospects.

Improved Performance of All-Solid-State Flexible Supercapacitor Based on the Stress-Compensation Effect.

This work presents a facile strategy to develop a flexible polyaniline (PANI)-based supercapacitor (SC) with both high energy density and good capacitance retention. An electrode with a symmetrical sandwich-structured configuration (PANI/flexible porous support/PANI) is used as both working and counter electrodes for this supercapacitor. For a conventional electrode with PANI depositing on single side of the support (PANI/flexible support), the flexible support bends severely during the PANI electrodeposition process, which results in poor PANI deposition. On the contrary, for the symmetrical sandwich-structured electrode, due to the stress-compensation effect induced by this configuration, the support bending is significantly suppressed and thus PANI films with a good uniformity are realized. Moreover, the stress-compensation effect involved in the symmetrical sandwichstructured electrode can also effectively balance the stress caused by PANI expansion/shrinkage during its electrochemical charge/discharge operation, thus improving the mechanical stability. The symmetrical sandwich-structured electrode has larger PANI mass loading, better PANI morphologies and stronger mechanical stability than those of the conventional electrode. Consequently, the SC constructed by the symmetrical sandwich-structured electrode displays better electrochemical performance in terms of its larger specific areal capacitance (369.2 mF·cm-2 at a current density of 0.25 mA·cm-2), higher energy density (0.031 mWh·cm-2 at a power density of 1.21 mW·cm-2) and better cycling retention (93.2% of the retained capacity over 6000 cycles) than the SC constructed by the conventional electrode.

[Dynamic maternal measles antibody level in infants: a longitudinal study].

OBJECTIVE: To understand the dynamic maternal measles antibody level in infants in Guangzhou. METHOD: Women who delivered babies in a maternal and child health care hospital in Guangzhou and their infants who were never affected by measles were included in our study. All the serum samples collected from mothers and infants were tested by using enzyme linked immunosorbent assay(ELISA)for the quantitative measurement of measles IgG antibodies. A longitudinal sero-epidemiological study was conducted to investigate the duration of the presence of measles maternal antibodies in infants based on the measurement at different time points(0, 3, 5, 7 month). RESULT: A total of 689 mothers and their 691 infants(two pairs of twins)were included in the study. The concentration and the sero-positivity rate of measles antibody in mothers was 513.8 mIU/ml and 81.6% respectively. The concentration and the sero-positivity rate of measles antibody in infants was 732.8 mIU/ml and 87.3%, respectively. The measles antibody levels of newborns were positively associated with those of mothers(r=0.917 5, P<0.001), which declined rapidly at 3 months after birth and mostly turned to be negative at 7 months after birth. Measles antibody turned be negative at 3 months after birth in low, middle level groups, but remained to be positive in high level group at 5 months after birth. CONCLUSION: The maternal measles antibody could rarely protect infants at age of 8 months even though mother had been vaccinated or acquired natural immunity. It is suggested to adjust the time of the first dose measles vaccination in infants and increase the measles antibody level of child bearing aged women to reduce the incidence of measles in infants.

[Alteration of cocaine, morphine and nicotine levels in serum of opiomaniacs treated by jie du ling].

20 Opiomaniacs were treated by Jie Du Ling consisted of some Chinese herbal medicine. Cocaine, morphine and nicotine levels in serum of 20 cases before and after treatment and 20 subjects for control were measured simultaneously by means of radioimmunoassay. The results showed that cocaine, morphine and nicotine levels in patients serum before treatment were significantly higher than that of control (P < 0.001), while the level of above-mentioned 3 substances in patient's serum after treatment were significantly lower than that of before treatment (P < 0.001). It is proved that Chinese medicinal herbs could significantly reduce the levels of cocaine, morphine and nicotine after treatment in clinical practice. It offered evidence for Chinese herbal medicine in stopping opiomania.