A Versatile Strategy of Designing Gold Nanoparticle-cDNA Nanoprobes in Developing Aptamer-Based Lateral Flow Assay for Small-Molecule Detection.

Aptamer-based lateral flow assay (Apt-LFA) has shown promising applications for small-molecule detection. However, the design of the AuNP (gold nanoparticle)-cDNA (complementary DNA) nanoprobe is still a big challenge due to the moderate affinity of the aptamer to small molecules. Herein, we report a versatile strategy to design a AuNPs@polyA-cDNA (poly A, a repeat sequence with 15 A bases) nanoprobe for small-molecule Apt-LFA. The AuNPs@polyA-cDNA nanoprobe contains a polyA anchor blocker, complementary DNA segment to DNA on the control line (cDNAc), partial complementary DNA segment with aptamer (cDNAa), and auxiliary hybridization DNA segment (auxDNA). Using adenosine 5'-triphosphate (ATP) as a model target, we optimized the length of auxDNA and cDNAa and achieved a sensitive detection of ATP. In addition, kanamycin was used as a model target to verify the universality of the concept. Therefore, this strategy can be easily extended to other small molecules; therefore, high application potential in Apt-LFAs can be envisaged.

The protective effects of yellow tea extract against loperamide-induced constipation in mice.

Yellow tea, a rare type tea from China, has a rich breadth of functional ingredients and benefits the gastrointestinal tract. However, it is not clear whether the yellow tea extract can alleviate constipation. Therefore, we used loperamide-induced constipation in mice to evaluate the effects of yellow tea extract. Fifty Kunming mice were randomly divided into five groups: normal, model, low-dose yellow tea extract, low-dose yellow tea extract prevention group, and high-dose yellow tea extract prevention group. Mice were administered yellow tea extract for 5 weeks followed by loperamide-induced constipation for the final 2 weeks. The results showed that yellow tea extract alleviated constipation symptoms by improving the fecal water content, defecation weight, and gastrointestinal transit rate. Yellow tea extract intervention also protected colon tissue, regulated serum neurotransmitters, and decreased the vasoactive intestinal peptide level. Furthermore, qRT-PCR indicated that yellow tea extract regulated genes associated with the constipation state, raised 5-HT3 and 5-HT4 and reduced AQP3 and AQP4 mRNA expression. Moreover, we found that yellow tea extract changed the gut microbiota composition. Community diversity and richness were increased and principal co-ordinate analysis demonstrated that the yellow tea extract prophylaxis groups differed from the model group. Difference analysis indicated that yellow tea extract increased Roseburia, Lachnospiraceae_UCG-006, and Bifidobacterium and decreased norank_f_Clostridiales_vadinBB60_group, unclassified_o_Bacteroidales, and Bacteroides, which are correlated with constipation. Based on these results, we believe that regular yellow tea consumption can effectively alleviate constipation.

Transcriptome analysis of the tea oil camellia (Camellia oleifera) reveals candidate drought stress genes.

BACKGROUND: The tea-oil camellia (Camellia oleifera) is the most important oil plant in southern China, and has a strong resistance to drought and barren soil. Understanding the molecular mechanisms of drought tolerance would greatly promote its cultivation and molecular breeding. RESULTS: In total, we obtained 76,585 unigenes with an average length of 810 bp and an N50 of 1,092 bp. We mapped all the unigenes to the NCBI 'nr' (non-redundant), SwissProt, KEGG, and clusters of orthologous groups (COG) databases, where 52,531 (68.6%) unigenes were functionally annotated. According to the annotation, 46,171 (60.8%) unigenes belong to 338 KEGG pathways. We identified a series of unigenes that are related to the synthesis and regulation of abscisic acid (ABA), the activity of protective enzymes, vitamin B6 metabolism, the metabolism of osmolytes, and pathways related to the biosynthesis of secondary metabolites. After exposed to drought for 12 hours, the number of differentially-expressed genes (DEGs) between treated plants and control plants increased in the G4 cultivar, while there was no significant increase in the drought-tolerant C3 cultivar. DEGs associated with drought stress responsive pathways were identified by KEGG pathway enrichment analysis. Moreover, we found 789 DEGs related to transcription factors. Finally, according to the results of qRT-PCR, the expression levels of the 20 unigenes tested were consistent with the results of next-generation sequencing. CONCLUSIONS: In the present study, we identified a large set of cDNA unigenes from C. oleifera annotated using public databases. Further studies of DEGs involved in metabolic pathways related to drought stress and transcription will facilitate the discovery of novel genes involved in resistance to drought stress in this commercially important plant.

Modification of lily polysaccharide by selenylation and the immune-enhancing activity.

Lily polysaccharide (LP) was extracted, purified and selenizingly modified by HNO3-Na2SeO3 method according to L9(3(4)) orthogonal design. Nine selenizing LPs, sLP1-sLP9, were obtained and their immune-enhancing activities were compared taking unmodified LP as control. The results in vitro test showed that sLP6 presented the strongest activity in promoting lymphocytes proliferation in single and synergetic with PHA, and the relative expression level of IL-2, IL-6 and IFN-γ mRNA of chicken peripheral lymphocytes. The results in vivo test showed that sLP6 could promote lymphocytes proliferation and enhance the serum antibody titers and serum IL-2, IL-6, IFN-γ contents more significantly than LP in chickens vaccinated with Newcastle Disease (ND) vaccine. These results indicate that polysaccharide selenizing can significantly enhance the immune-enhancing activity of LP and the optimal modification conditions are 400 mg of Na2SeO3 per 500 mg of LP, the reaction temperature of 70 °C and the reaction time of 6 h.

Optimization of selenylation modification for garlic polysaccharide based on immune-enhancing activity.

Garlic polysaccharide (GPS) was modified in selenylation respectively by nitric acid-sodium selenite (NA-SS), glacial acetic acid-selenous acid (GA-SA), glacial acetic acid-sodium selenite (GA-SS) and selenium oxychloride (SOC) methods each under nine modification conditions of L9(3(4)) orthogonal design and each to obtain nine selenizing GPSs (sGPSs). Their structures were identified, yields and selenium contents were determined, selenium yields were calculated, and the immune-enhancing activities of four sGPSs with higher selenium yields were compared taking unmodified GPS as control. The results showed that among four methods the selenylation efficiency of NA-SS method were the highest, the activity of sGPS5 was the strongest and significantly stronger than that of unmodified GPS. This indicates that selenylation modification can significantly enhance the immune-enhancing activity of GPS, NA-SS method is the best method and the optimal conditions are 0.8:1 weight ratio of sodium selenite to GPS, reaction temperature of 70 °C and reaction time of 10h.

Effects of Selenylation Modification on Antioxidative Activities of Schisandra chinensis Polysaccharide.

The selenylation modification of Schisandra chinensis polysaccharide (SCP) was conducted by the HNO3-Na2SeO3 method respectively under nine conditions according to L9(34) orthogonal design. Nine selenizing SCPs, sSCP1-sSCP9, were obtained, and their antioxidant activities were compared. In vitro test, the free radical-scavenging rates of nine sSCPs were determined for DPPH., .OH and ABTS+. sSCP1 presented the most significant effect, and could inhibit the nonenzymatic protein glycation. In vivo test, 14-day-old chickens were injected respectively with sSCP1 and SCP, the serum contents of CAT, SOD and MDA were determined. The result showed that as compared with the SCP group, the SOD and CAT activities were significantly or numerically raised and MDA content was significantly or numerically lowered in the sSCP1 group. These results indicate that selenylation modification can significantly enhance the antioxidant and antiglycative activity of SCP in vitro or in vivo. sSCP1 possesses the best efficacy and its modification conditions can be as optimal modification conditions that were 200 mg of Na2SeO3 for 500 mg of SCP, reaction temperature of 50°C and reaction time of 6 h.

Effects of selenizing angelica polysaccharide and selenizing garlic polysaccharide on immune function of murine peritoneal macrophage.

The effects of two selenizing polysaccharides (sCAP2 and sGPS6) on immune function of murine peritoneal macrophages taking two non-selenizing polysaccharides (CAP and GPS) and modifier Na2SeO3 as control. In vitro test, the changes of selenizing polysaccharides, non-selenizing polysaccharides and Na2SeO3 on murine macrophages function were evaluated by phagocytosis and nitric oxide (NO) secretion tests. In vivo test, the mice were injected respectively with 0.2, 0.4 and 0.6 mg of sCAP2, sGPS6, CAP and GPS, or Na2SeO3 80 µg or normal saline 0.4 mL. The peritoneal macrophages were collected and cultured to determine the contents of TNF-α, IL-6 and IL-10 in supernatants by enzyme-linked immunosorbent assay. The results showed that sCAP2 and sGPS6 could significantly promote the phagocytosis and secretion of NO and three cytokines of macrophages in comparison with CAP and GPS. sCAP2 possessed the strongest activity. This indicates that selenylation modification can further improve the immune-enhancing activity of polysaccharide, and sCAP2 could be as a new immunopotentiator.

Effect of selenylation modification on immune-enhancing activity of Atractylodes macrocephala polysaccharide.

The Atractylodes macrocephala polysaccharide (AMP) was extracted purified and modified in selenylation by Nitric acid-sodium selenite method to get nine selenizing AMPs (sAMPs), sAMP(1)-sAMP(9). In vitro test their effects on chicken peripheral lymphocyte proliferation were determined by MTT assay. The results showed that nine sAMPs and AMP at five concentrations could significantly promote lymphocyte proliferation, the actions in six sAMPs were significantly stronger than that in AMP, and in sAMP(9) was the strongest. In vivo test, 14-day-old chickens vaccinated with ND vaccine were injected respectively with sAMP(9) and AMP, the peripheral lymphocytes proliferation, serum antibody titer, IFN-γ, IL-2 and IL-6 contents were determined. The results displayed that the sAMP could significantly promote lymphocyte proliferation and elevate the antibody titers and content of IFN-γ, IL-2 and IL-6 in comparison with unmodified AMP. These results indicate that selenylation modification can significantly enhance the immune-enhancing activity of AMP.

Optimization of selenylation conditions for lycium barbarum polysaccharide based on antioxidant activity.

Lycium barbarum polysaccharide (LBP) was modified by HNO3-Na2SeO3 method according to L9(3(4)) orthogonal design to obtain nine selenizing LBPs (sLBPs), sLBP1-sLBP9. Their antioxidant activities in vitro were compared by free radical-scavenging test. sLBP6, sLBP8 and sLBP9 presented stronger activity. In vivo test, 14-day-old chickens were injected respectively with sLBP6, sLBP8 and sLBP9 taking LBP as control, and serum GSH-Px and SOD activities and MDA content were determined. The results showed that three sLBPs could significantly enhance GSH-Px and SOD activities and decrease MDA content. The actions of sLBPs were significantly stronger than that of unmodified LBP. These results indicated that selenylation modification could significantly enhance the antioxidant activities of LBP, sLBP6 possessed the best efficacy and could be exploited into an antioxidant. The optimal modification conditions were 400mg of sodium selenite for 500 mg of LBP, reaction temperature of 70 °C and reaction time of 6h.

Effects of selenylation modification on immune-enhancing activity of garlic polysaccharide.

The garlic polysaccharide was modified by HNO3-Na2SeO3 method according to orthogonal design L9(3(4)) to obtain nine selenizing garlic polysaccharides, sGPS1-sGPS9. Their effects on chicken peripheral lymphocytes proliferation in vitro were compared by MTT assay. The results showed that sGPSs could significantly promote lymphocytes proliferation, sGPS3, sGPS5 and sGPS6 presented stronger efficacy. In vivo experiment, 14-day-old chickens were injected respectively with sGPS3, sGPS5 and sGPS6 when they were vaccinated with ND vaccine taking unmodified GPS as control. The results showed that three sGPSs could significantly promote lymphocyte proliferation, enhance serum antibody titer, IFN-γ and IL-2 contents. These results indicated that selenylation modification could significantly enhance the immune-enhancing activity of GPS, sGPS6 possessed the best efficacy and could be as a candidate drug of immunoenhancer. Its optimal modification conditions were 400 mg of sodium selenite for 500 mg of GPS, reaction temperature of 70°C and reaction time of 6 h.

[Multicentral randomized controlled studies on acupuncture at Shaoze (SI 1) for treatment of postpartum hypolactation].

OBJECTIVE: To explore effect of acupuncture at Shaoze (SI 1) for treatment of postpartum hypolactation. METHODS: Multicentral randomized controlled and single blind clinical trial was adopted, and 276 cases were divided into a treatment group and a control group, 138 cases in each group. The treatment group were treated with electroacupuncture (EA) at Shaoze (SI 1) and the control group with EA at Shangyang (LI 1). After treatment of 2 courses, the therapeutic effects and changes of cumulative score of TCM symptoms, mammary filling degree, lactation amount, prolactin level were evaluated and investigated. RESULTS: The cured and markedly effective rate was 97.8% in the treatment group and 24.3% in the control group with a significant difference between the two groups (P < 0.05). The treatment group in improvement of the cumulative score of TCM symptoms and the mammary filling degree, and increasing the lactation amount and the prolactin level were better than the control group (P < 0.01). CONCLUSION: Acupuncture at Shaoze (SI 1) has obvious therapeutic effect on hypolactation.

[The inhibitory effect of extracts from Galla chinensis on marine fouling bacteria].

OBJECTIVE: To observe the activity of extracts from Galla Chinensis with different solvents against marine fouling bacteria in vitro. METHOD: Incubating plate with foveolae was used. RESULTS: The ethyl acetate, ethanol and water extracts showed strong inhibitory activity against marine fouling bacteria belonging to nine genera including Pseudomonas, Aeromonas, Alealigenes, Flavobacterium, Vibrio, Photobacterium, Moraxella, Chromobacterium, Enterobacter. While, the petroleum ether and chloroform extracts showed no inhibitory activity. CONCLUSION: The study provided theory basis for new biologic protective dope on environment.