The practice of a mixed methods research strategy: personal, professional and project considerations

Effect of DE value on starch solubility of kudzu root starch

The Circular RNA Cdr1as Act as an Oncogene in Hepatocellular Carcinoma through Targeting miR-7 Expression

<div><p>CircRNAs are a class of endogenous RNA that regulates gene expression at the post-transcriptional or transcriptionallevel through interacting with other molecules or microRNAs. Increasing studies have demonstrated that circRNAs play a crucial role in biology processes. CircRNAs are proved as potentialbiomarkers in many diseases including cancers. However, the role of Cdr1as in Hepatocellular carcinoma (HCC) remains to be elucidated. We demonstrated that Cdr1as expression was upregulated in HCC tissues compared with the adjacent non-tumor tissues. In addtion, miR-7 expression was downregulated in HCC tissues compared with the adjacent non-tumor tissues. Moreover, the expression level of miR-7 was inversely correlated with that in HCC tissues. Knockdown of Cdr1as suppressed the HCC cell proliferation and invasion. Overexpression of miR-7 inhibited the HCC cell proliferation and invasion. Overexpression of miR-7 could suppress the direct target gene CCNE1 and PIK3CD expression. Knockdown of Cdr1as suppressed the expression of miR-7 and also inhibited the CCNE1 and PIK3CD expression. Furthermore, knockdown of Cdr1as suppressed the HCC cell proliferation and invasion through targeting miR-7. These data suggested that Cdr1as acted as an oncogene partly through targeting miR-7 in HCC.</p></div

Knockdown of Cdr1as suppressed the HCC cell proliferation and invasion through targeting miR-7 expression.

<p>(A) The expression of miR-7 in the SMMC-7721 cell was determined by qRT-PCR. (B) The expression of miR-7 was measured by qRT-PCRinthe SMMC-7721 cell. (C) Cell proliferation was detected by CCK-8 analysis in the SMMC-7721 cell. (D) The cell invasive ability was measured by invasion assay in the SMMC-7721 cell. *p<0.05, **p<0.01 and ***p<0.001.</p

Programming a Biofilm-Mediated Multienzyme-Assembly-Cascade System for the Biocatalytic Production of Glucosamine from Chitin

Chitin is used as an essential raw material for the production of glucosamine (GlcN). In this study, we adopted three key enzymes, isolated from <i>Thermococcus kodakaraensis</i> KOD1, that catalyze the sequential conversion of α-chitin into GlcN and developed a multienzyme-assembly-cascade (MAC) system immobilized in a bacterial biofilm, which enabled a multistep one-pot reaction. Specifically, the SpyTag–SpyCatcher and SnoopTag–SnoopCatcher pairs provided covalent and specific binding force to fix enzymes to the biofilm one by one and assemble close enzyme cascades. The MAC system showed great catalytic activity, converting 79.02 ± 3.61% of α-chitin into GlcN with little byproducts, which was 2.09 times that of GlcN catalyzed by a mixture of pure enzymes. The system also exhibited good temperature and pH stability. Notably, 90% of enzyme activity was retained after 6 rounds of reuse, and appreciable activity remained after 17 rounds

The expression of Cdr1as was upregulated in the HCC tissues and was inverse associated with the expression of miR-7.

<p>(A) The expression of Cdr1as was detected in the HCC tissues and adjacent non-tumor tissues using qRT-PCR. (B) Cdr1as expression was upregulated in the (74%, 26/35) HCC tissues compared with their adjacent non-tumor tissues. (C) The expression of miR-7 was detected in the HCC tissues and adjacent non-tumor tissues using qRT-PCR. (D) miR-7 expression was downregulated in the (66%, 23/35) HCC tissues compared with their adjacent non-tumor tissues. (E) The expression of miR-7 was inversely correlated with the expression of Cdr1as in the HCC tissues.</p

Knockdown of Cdr1as promoted the miR-7 and its target gene CCNE1 and PIK3CD expression.

<p>(A) The mRNA expression of miR-7 in the SMMC-7721 cell was measured by qRT-PCR. (B) The mRNA expression of CCNE1 was detected by qRT-PCR in the SMMC-7721 cell. (C) The protein expression of CCNE1 was detected by western blot. (D) The mRNA expression of CCNE1 in the SMMC-7721 cell was detected by qRT-PCR. (E) The protein expression of CCNE1 was detected by western blot. (F) The mRNA expression of PIK3CD was detected by qRT-PCR in the SMMC-7721 cell. (G) The protein expression of PIK3CD was measured using western blot. (H) The mRNA expression of PIK3CD was detected by qRT-PCR in the SMMC-7721 cell. (J) The protein expression of PIK3CD was determined by western blot.</p

MicroRNA-424 Is Down-Regulated in Hepatocellular Carcinoma and Suppresses Cell Migration and Invasion through c-Myb

<div><p>Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality worldwide. MicroRNAs (miRNAs) are important regulators of multiple cellular processes, and the aberrant miRNAs expressions have been observed in different types of cancer including HCC. Their pathysiologic role and their relevance to tumorigenesis are still largely unknown. In this study, we demonstrated the down-regulation of miR-424 in HCC cell lines and tissues by quantitative RT-PCR analyses. Overexpression of miR-424 reduced the HCC cell prolifetation, migration, and invasion. Conversely, inhibiton of miR-424 expression significantly accelerated the cell proliferation, migration, and invasion. In addition, we further identified c-Myb as a functional downstream target of miR-424 by directly targeting the 3′UTR of c-Myb. Furthermore, overexpression of c-Myb impaired miR-424-induced inhibition of proliferation and invasion in HCC cells. Our results demonstrated that miR-424 was involved in tumorigenesis of HCC at least in part by suppression of c-Myb.</p></div

Additional file 1: Figure S1. of Optimization of bioconversion process for trehalose production from enzymatic hydrolysis of kudzu root starch using a visualization method

Principles of visualization method

Knockdown of Cdr1as suppressed the HCC cell proliferation and invasion.

<p>(A) The expression of Cdr1as in the HCC cell lines were measured by using qRT-PCR. (B) Knockdown of Cdr1as expression could suppress the Cdr1as expression. (C) Knockdown of Cdr1as expression inhibited the HCC cell line (SMMC-7721) proliferation. (D) The HepG2 cell proliferation was measured by CCK-8 assay. (E) Knockdown of Cdr1as expression inhibited the SMMC-7721 cell invasion. (F) The HepG2 cell invasion was detected by cell invasion assay. *p<0.05, **p<0.01 and ***p<0.001.</p

Overexpression of c-Myb impairs miR-424-induced inhibition of proliferation and invasion in HCC cells.

<p>(A) Western blot analysis of c-Myb expression in 8 miR-424 down-regulated HCC tissues. GAPDH was also detected as a loading control. (B) Western blot analysis of c-Myb expression in HepG2 cells co-transfected with either miR-424 mimics or scramble and pCDNA-c-Myb or pCDNA empty vector; GAPDH was also detected as a loading control. (C) Cell growth curves in HepG2 cells transfected with different combinations. (D) Transwell analysis of HepG2 cells treated with different combinations. The relative ratio of invasive cells per field is shown right. *p<0.05,** p<0.01, ***p<0.001.</p