Selection and validation of reference genes for RT-qPCR analysis in Sf9 cell line infected by Heliothis virescens ascovirus 3h (HvAV-3h)

Zi-Shu Chen,Ning-Ning Han,Jian Hong Li,Hu Wan,Guo-Hua Huang 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.2

The real-time quantitative reverse transcription PCR (qRT-PCR) is an efficient and pervasive technique for analysis gene expression. Despite the fact that this technique has been extensively used to explore the gene functions, the stability of the reference genes still requires being validated at different experimental conditions. This research aims to confirm that the expression stability of seven candidates reference genes under different concentrations sterile hemolymph containing virus (2, 4, 6, 108-fold) infects the Spodoptera frugiperda cells line (Sf9). The appropriate reference genes as endogenous controls were determined through four computational algorithms (ΔCt, NormFinder, BestKeeper, and geNorm) and an integrated online software RefFinder. According to results of this study, TUB and ACT1 were proved to be the most stable reference genes in E2 fold and E4 fold respectively. PPI was the best reference gene in E6 and E8 fold, 28S and TUB were the most stable pair reference genes through all samples. These results can facilitate basic researches of the molecular mechanism of HvAV- 3h and also prove to be helpful for the standard RT-PCR method.

Optimizing Preparation Conditions for Angiotensin-I-Converting Enzyme Inhibitory Peptides Derived from Enzymatic Hydrolysates of Ovalbumin

Qun Huang,Shu-gang Li,Hui Teng,Yong-guo Jin,Mei-hu Ma,Hong-bo Song 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.6

Angiotensin-I-converting enzyme (ACE) inhibitory peptides were prepared from ovalbumin using enzyme hydrolysis with pepsin as an enzyme source. Effects of pH, enzyme dosage, substrate concentration, hydrolysis temperature, and time on the degree of hydrolysis and the ACE inhibition rate were investigated using single factor experiments. Preparation conditions for ACE inhibitory peptides were optimized using a response surface design on the base of single factor experiments. Optimum preparation conditions were a substrate concentration of 5.2 g/100 mL of D.W with a pH value of 2.5, an enzyme dosage of 14,000 U/g, and a hydrolysis time of 250 min at 30℃. The ACE inhibition rate was up to 70.55±1.13% under these conditions.

Analysis of the Involvement of Chitin-Binding Domain of ChiCW in Antifungal Activity, and Engineering a Novel Chimeric Chitinase with High Enzyme and Antifungal Activities

Chien Jui Huang,Shu Huei Guo,Shu Chun Chung,Yu Ju Lin,Chao Ying Chen 한국미생물 · 생명공학회 2009 Journal of microbiology and biotechnology Vol.19 No.10

Evaluation of storage period of fresh ginseng for quality improvement of dried and red processed varieties

Na Zhang,Xin Huang,Yun-Long Guo,Hao Yue,Chang-Bao Chen,Shu-Ying Liu 고려인삼학회 2022 Journal of Ginseng Research Vol.46 No.2

Background: Dried and red ginseng are well-known types of processed ginseng and are widely used as healthy food. The dried and red ginseng quality may vary with the storage period of raw ginseng. Therefore, herein, the effect of the storage period of fresh ginseng on processed ginseng quality was evaluated through multicomponent quantification with statistical analysis. Methods: A method based on ultrahigh performance liquid chromatography coupled to triple quadrupole mass spectrometry in multiple-reaction monitoring mode (UPLC-MRM-MS) was developed for quantitation of ginsenosides and oligosaccharides in dried and red ginseng. Principal component analysis and partial least squares discriminant analysis were conducted to evaluate the dynamic distributions of ginsenosides and oligosaccharides after different storage periods. Results: Eighteen PPD, PPT and OLE ginsenosides and nine reducing and nonreducing oligosaccharides were identified and quantified. With storage period extension, the ginsenoside content in the processed ginseng increased slightly in the first 2 weeks and decreased gradually in the following 9 weeks. The content of reducing oligosaccharides decreased continuously as storage time extending, while that of the nonreducing oligosaccharides increased. Chemical conversions occurred during storage, based on which potential chemical markers for the storage period evaluation of fresh ginseng were screened. Conclusion: According to ginsenoside and oligosaccharide distributions, it was found that the optimal storage period was 2 weeks and that the storage period of fresh ginseng should not exceed 4 weeks at 0 °C. This study provides deep insights into the quality control of processed ginseng and comprehensive factors for storage of raw ginseng.

MR Imaging of the Spine at 3.0T with T2-Weighted IDEAL Fast Recovery Fast Spin-Echo Technique

Ai-Jun Ren,Yong Guo,Shu-Ping Tian,Li-Jing Shi,Min-Hua Huang 대한영상의학회 2012 Korean Journal of Radiology Vol.13 No.1

Objective: To compare the iterative decomposition of water and fat with echo asymmetry and the least-squares estimation (IDEAL) method with a fat-saturated T2-weighted (T2W) fast recovery fast spin-echo (FRFSE) imaging of the spine. Materials and Methods: Images acquired at 3.0 Tesla (T) in 35 patients with different spine lesions using fat-saturated T2W FRFSE imaging were compared with T2W IDEAL FRFSE images. Signal-to-noise ratio (SNR)-efficiencies measurements were made in the vertebral bodies and spinal cord in the mid-sagittal plane or nearest to the mid-sagittal plane. Images were scored with the consensus of two experienced radiologists on a four-point grading scale for fat suppression and overall image quality. Statistical analysis of SNR-efficiency, fat suppression and image quality scores was performed with a paired Student’s t test and Wilcoxon’s signed rank test. Results: Signal-to-noise ratio-efficiency for both vertebral body and spinal cord was higher with T2W IDEAL FRFSE imaging (p < 0.05) than with T2W FRFSE imaging. T2W IDEAL FRFSE demonstrated superior fat suppression (p < 0.01) and image quality (p < 0.01) compared to fat-saturated T2W FRFSE. Conclusion: As compared with fat-saturated T2W FRFSE, IDEAL can provide a higher image quality, higher SNR-efficiency, and consistent, robust and uniform fat suppression. T2W IDEAL FRFSE is a promising technique for MR imaging of the spine at 3.0T. Objective: To compare the iterative decomposition of water and fat with echo asymmetry and the least-squares estimation (IDEAL) method with a fat-saturated T2-weighted (T2W) fast recovery fast spin-echo (FRFSE) imaging of the spine. Materials and Methods: Images acquired at 3.0 Tesla (T) in 35 patients with different spine lesions using fat-saturated T2W FRFSE imaging were compared with T2W IDEAL FRFSE images. Signal-to-noise ratio (SNR)-efficiencies measurements were made in the vertebral bodies and spinal cord in the mid-sagittal plane or nearest to the mid-sagittal plane. Images were scored with the consensus of two experienced radiologists on a four-point grading scale for fat suppression and overall image quality. Statistical analysis of SNR-efficiency, fat suppression and image quality scores was performed with a paired Student’s t test and Wilcoxon’s signed rank test. Results: Signal-to-noise ratio-efficiency for both vertebral body and spinal cord was higher with T2W IDEAL FRFSE imaging (p < 0.05) than with T2W FRFSE imaging. T2W IDEAL FRFSE demonstrated superior fat suppression (p < 0.01) and image quality (p < 0.01) compared to fat-saturated T2W FRFSE. Conclusion: As compared with fat-saturated T2W FRFSE, IDEAL can provide a higher image quality, higher SNR-efficiency, and consistent, robust and uniform fat suppression. T2W IDEAL FRFSE is a promising technique for MR imaging of the spine at 3.0T.

The High Expressed Serum Soluble Neural Cell Adhesion Molecule, a High Risk Factor Indicating Hepatic Encephalopathy in Hepatocelular Carcinoma Patients

Liu, Tian-Hua,Guo, Kun,Liu, Ri-Qiang,Zhang, Shu,Huang, Zhuo-Hui,Liu, Yin-Kun Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.8

Objective: To investigate whether the expression of serum soluble neural cell adhesion molecule (sNCAM) is associated with hepatic encephalopathy (HE) in hepatocelular carcinoma (HCC) patients. Materials and Methods: The Oncomine Cancer Microarray database was used to determine the clinical relevance of NCAM expression in different kinds of human cancers. Sera from 75 HCC cases enrolled in this study were assessed for expression of sNCAM by enzyme linked immunosorbent assay (ELISA). Results: Dependent on the Oncomine Cancer Microarray database analysis, NCAM was down regulated in 10 different kinds of cancer, like bladder cancer, brain and central nervous system cancer, while up-regulated in lung cancer, uterine corpus leiomyoma and sarcoma, compared to normal groups. Puzzlingly, NCAM expression demonstrated no significant difference between normal and HCC groups. However, we found by quantitative ELISA that the level of sNCAM in sera from HCC patients with HE ($347.4{\pm}151.9ng/ml$) was significantly more up-regulated than that in HCC patients without HE ($260.3{\pm}104.2ng/ml$), the p-value being 0.008. sNCAM may be an important risk factor of HE in HCC patients, the correlation coefficients was 0.278 (P<0.05) on rank correlation analysis. Conclusions: This study highlights that up-regulated level of serum sNCAM is associated with HE in HCC patients and suggests that the high expression can be used as an indicator.

UHPLC–HRMS based saponins profiling of three morphological regions in American ginseng (Panax quinquefolium L.) and their correlation with the antioxidant activity

Yuxin Bai,Guangzhi Cai,Na Guo,Xin Huang,Jiyu Gong,Shu-Ying Liu,Yun-Long Guo,Wei Wang 한국식품과학회 2024 Food Science and Biotechnology Vol.33 No.7

American ginseng (Panax quinquefolium L.) is used as tonic plant and high-grade nourishment. Ultra-high-performance liquid chromatography–high resolution mass spectrometry (UHPLC–HRMS) method was established for identifying the chemical constituent in three morphological regions of American ginseng, including main root (MR), rhizome (RH) and lateral root (LR). The 63 saponins was identified in different morphological regions of 10 American ginseng samples. The chemical maker compounds in corresponding morphological region, while the major compounds of MR (malonyl-ginsenoside Rb1, ginsenoside Rd, Rs2 and pseudo-RC1), LR (stipuleanoside R2, ginsenoside Re and malonyl-ginsenoside Rc), and RH (malonyl-ginsenoside Rd, Rb3, and chikusetsu saponin II) were discovered. Correlation analysis showed that 11 compounds were positively correlated with the antioxidant activity of American ginseng.

Mesenchymal Stem Cells on a Decellularized Cartilage Matrix for Cartilage Tissue Engineering

Xi-Fu Zheng,Shi-Bi Lu,Wei-Guo Zhang,Shu-Yun Liu,Jing-Xiang Huang,Quan-Yi Guo 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.3

An ideal scaffold for cartilage tissue engineering should be biomimetic in not only its biochemical composition, but also in the morphological structure of the scaffold. In this study, we fabricated a scaffold with an oriented structure using a nanofibrous articular cartilage extracellular matrix (ACECM), in which the ACECM was used to mimic the biochemical composition and oriented structure of articular cartilage. Histology analysis showed that the scaffold contained cartilage ECM (GAGs and collagen II). Scanning electron microscopy (SEM) indicated that the scaffolds were composed of nanofibers and possessed vertical microtubules. Chondrogenic differentiation-induced mesenchymal stem cells (MSCs) were seeded on the scaffold in vitro. SEM showed that MSCs proliferated well and aligned along the vertical microtubules,which mimicked the orientation of deep zone articular cartilage. A cell proliferation assay and live/dead cell staining demonstrated that the ACECM possessed good cell affinity, which favored cell adherence and proliferation. The MSCs that had been labeled with the fluorescent dye PKH26 and seeded on scaffolds were implanted into nude mice. The differentiated cells/ACECM implants formed cartilage-like tissue 4 weeks after implantation, and stained positive for collagen type II and toluidine blue. In addition,the in vivo fluorescent images verified that the MSCs in the implants were the labeled MSCs. These results demonstrated that the oriented ACECM scaffolds hold great promise for use in cartilage tissue engineering applications.

Survival Analysis of Oral Squamous Cell Carcinoma in a Subgroup of Young Patients

Fan, Yi,Zheng, Lei,Mao, Ming-Hui,Huang, Ming-Wei,Liu, Shu-Ming,Zhang, Jie,Li, Sheng-Lin,Zheng, Lei,Zhang, Jian-Guo Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.20

Oral squamous cell carcinoma (OSCC) is predominantly a disease of middle-aged men with long-term exposure to tobacco and alcohol. An increasing trend has been reported at a younger age worldwide. Clinical records of 100 patients under the age of 45 years treated specifically for oral cavity SCC in our hospital during a 10-year period were retrospectively analyzed to calculate the survival rates. An obvious male predominance coincided with smoking trend among Chinese young individuals and female patients were more likely to have no traditional risk factors such as smoking or drinking. The 5-year overall survival rate and disease-free survival rate were 61.0% and 75.5%, respectively, consistent with other published series over the decade showing a relatively better survival among the young. No significant differences clearly correlated with outcome when comparing non-smokers non-drinkers to ever-smokers and ever drinkers (P>0.05). Overall survival rate and disease free survival rate was found to be significantly higher in patients with early-stage disease than with advanced stage disease (P=0.001, P=0.009 respectively). The strong influence of clinical stage on prognosis emphasizes the importance of early diagnosis and treatment of oral malignancies for this unique clinical subgroup.

Circular RNA cFAM210A, degradable by HBx, inhibits HCC tumorigenesis by suppressing YBX1 transactivation

Yu Jian,Li Wen,Hou Guo-jun,Sun Da-peng,Yang Yuan,Yuan Sheng-xian,Dai Zhi-hui,Yin Hao-zan,Sun Shu-han,Huang Gang,Zhou Wei-ping,Yang Fu 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Hepatitis B protein x (HBx) has been reported to promote tumorigenesis in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), but the mechanism awaits further investigation. In this study, we found that cFAM210A (a circular RNA derived from the third exon of transcript NM_001098801 of the FAM210A gene; CircBase ID: hsa_circ_0003979) can be silenced by HBx. cFAM210A expression was downregulated and negatively correlated with tumorigenesis in patients with HBV-related HCC. Furthermore, cFAM210A reduced the proliferation, stemness, and tumorigenicity of HCC cells. Mechanistically, HBx increased the N6-methyladenosine (m6A) level of cFAM210A by promoting the expression of RBM15 (an m6A methyltransferase), thus inducing the degradation of cFAM210A via the YTHDF2-HRSP12-RNase P/MRP pathway. cFAM210A bound to YBX1 and inhibited its phosphorylation, suppressing its transactivation function toward MET. These findings suggest the important role of circular RNAs in HBx-induced hepatocarcinogenesis and identify cFAM210A a potential target in the prevention and treatment of HBV-related HCC.