• International Journal of Industrial Entomology and Biomaterials
• /
• v.12 no.1
• /
• pp.15-19
• /
• 2006

RNA interference has been used as a powerful tool in preventing virus proliferation in many species. In this study, we injected the dsRNA in vitro transcripts into Bombyx mori to investigate the resistance to B. mori nuclear polyhedrosis virus (BmNPV). Through vivisectional observation and real-time quantities PCR analysis, we found that these dsRNA can prevent the BmNPV to a certain extent, and delay the viruses' proliferation.

• Food Science of Animal Resources
• /
• v.29 no.4
• /
• pp.506-512
• /
• 2009

Salmonellosis is an important worldwide foodborne infectious disease that is transmitted by many food vehicles including raw and processed animal products and fresh produce. In this study, the effectiveness of automated ELISA ($VIDAS^{(R)}$) and realtime PCR in the detection of Salmonella spp. in steamed pork and raw broccoli sprouts was evaluated by comparing their results with those of a conventional culture method. Bulk samples (500 g) of steamed pork and raw broccoli sprouts were inoculated with various levels of Salmonella and divided into 20 samples (25 g each). All the samples, including the controls, were analyzed using a conventional culture method, $VIDAS^{(R)}$, and real-time PCR to detect the presence of Salmonella. In addition, the levels of background flora in the steamed pork and the raw broccoli sprouts were determined. In the steamed pork that contained less than 100 CFU/g of aerobic bacteria, all three methods detected low levels of Salmonella without a statistical difference in their performance. In the broccoli sprouts with high quantities of background flora (ca. $6.7{\times}10^7$ CFU/g), however, all three methods were unable to detect low levels of Salmonella, and real-time PCR and $VIDAS^{(R)}$ more sensitively detected Salmonella than the culture method, with significant statistical differences. In conclusion, $VIDAS^{(R)}$ and real-time PCR could be superior to conventional culture methods in detecting Salmonella in food with high levels of background flora.

• Microbiology and Biotechnology Letters
• /
• v.48 no.4
• /
• pp.447-454
• /
• 2020

Veillonella spp. have been reported to be the most prevalent nitrate-reducing bacterial species in the oral cavity. The purpose of this study was to examine the relationship between the abundance of Veillonella spp. and nitrite production after nitrate ingestion. Bacterial samples were obtained from the tongue surfaces of 50 university students. The predominant Veillonella spp., V. atypica, V. dispar, and V. rogosae were identified and enumerated using real-time polymerase chain reaction (qPCR). Salivary nitrate and nitrite were measured before and 30, 60, and 90 min after ingestion of 100 ml of beetroot juice. Increased nitrite concentrations were observed in all participants, with a mean increase of 0.61 (0.42-1.10) mM expressed as the median (interquartile range). Veillonella atypica was detected in 40 subjects (80%), V. dispar in 48 (96%), and V. rogosae in 48 (96%), at quantities ranging from 1.3 × 102 to 2.8 × 107 CFU/ml per subject. The strengths of the correlations of the log colony forming unit (CFU) values of V. atypica, V. dispar, V. rogosae, and the log CFU value of the three species together with the increase in nitrite levels were 0.091, 0.114, -0.228, and 0.060, respectively, none of which were significant (p > 0.05). Our results indicate that the abundance of Veillonella spp. is not related to salivary nitrite production after nitrate ingestion.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.5
• /
• pp.2987-2990
• /
• 2013

Background: To explore mRNA expression and clinical significance of ERCC1, BRCA1, RRM1, TYMS and TUBB3 genes in tumor tissue of postoperative patients with non-small cell lung cancer (NSCLC). Materials and Methods: Sixty NSCLC patients undergoing radical operation in our hospital from Nov., 2011 to Jun., 2012 were selected. Plasmid standards of ERCC1, BRCA1, RRM1, TYMS and TUBB3 were established and standard curves were prepared by SYBR fluorescent real-time quantitative PCR analysis. Samples from tumor centers were taken to detect mRNA expression of ERCC1, BRCA1, RRM1, TYMS and TUBB3 genes in cancerous tissue during operation. The total mRNA expression quantities were compared according to different clinical characteristics. Results: The total expression quantities of 5 genotypes from high to low were ERCC1>RRM1>TUBB3>TYMS>BRCA1 in turn. By pairwise comparisons, other differences showed statistical significance (p<0.05 or p<0.01) except for TYMS and TUBB3 (p>0.05); the low expression rates from high to low were ERCC1>TYMS>TUBB3>TUBB3>RRM1>BRCA1 in turn. The expression quantities of BRCA1, RRM1 and TYMS in males, smokers and patients without adenocarcinoma were all significantly higher than that in females, non-smokers and patients with adenocarcinoma, and significant differences were present (p<0.05 or p<0.01). In terms of pathological staging, the expression quantities of BRCA1, RRM1 and TYMS in phases IIa~IIb and IIIa~IIIb had a tendency to be greater than in phases I and IV. Conclusions: Resistance to chemotherapy and sensitivity to targeted therapy differ among patients with NSCLC. Differences in gene expression in different individuals were also revealed. Only according to personalized detection results can individualized therapeutic regimens be worked out, which is a new direction for oncotherapy.

• Horticultural Science & Technology
• /
• v.29 no.6
• /
• pp.623-632
• /
• 2011

To increase the anti-carcinogens phenylethylisothiocyanate (PEITC), myrosinase (MYR), and glutathione S-transferase (GST), genes related to PEITC pathway were isolated and the gene expressions were regulated by Agrobacterium transformation. Isolated cDNAs, MYR, and GST genes were 1,647 bp and 624 bp, respectively, and the protein expression was confirmed through pET system. Thereafter, we constructed a sense-oriented over-expressing myrosinase (pBMY) and RNAi down-regulated GST (pJJGST) binary vectors for the Chinese cabbage transformation. After the transformation, thirteen over-expressing transgenic Chinese cabbage plants (IMS) with pBMY and five down-regulated ones (IGA) with pJJGST were selected by PCR analysis. Selected $T_0$ transgenic plants were generated to $T_1$ plants by self-pollination. Based on the Southern blot analysis on these $T_1$ transgenic plants, 1-4 copies of T-DNA were transferred to Chinese cabbage genome. Thereafter, RNA expression level of myrosinase gene or GST gene was analyzed through real-time RT PCR of IMS, IGA, and non-transgenic inbred lines. In case of IMS lines, myrosinase gene was increased 1.03-4.25 fold and, in IGA lines, GST gene was decreased by 26.42-42.22 fold compared to non-transgenic ones, respectively. Analysis of PEITC concentrations using GC-MS it showed that some IMS lines and some IGA lines increased concentrations of PEITC up to 4.86 fold and up to 3.89 fold respectively compared to wild type. Finally in this study IMS 1, 3, 5, 12, and 15 and IGA 1, 2, and 4 were selected as developed transgenic lines with increasing quantities of anti-carcinogen PEITC.

• Journal of Nutrition and Health
• /
• v.50 no.5
• /
• pp.415-425
• /
• 2017

Purpose: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against $H_2O_2-induced$ cell death of human neuroblastoma SK-N-MC cells. Methods: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. Results: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The $IC_{50}$ value for DPPH radical scavenging activity was $506.95{\mu}g/ml$ for MCE. Pre-treatment with MCE showed protective effects against $H_2O_2-induced$ cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of $H_2O_2$. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by $H_2O_2$. Conclusion: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.