Study on characterization of Bupleurum chinense polysaccharides with antioxidant mechanisms focus on ROS relative signaling pathways and anti-aging evaluation in vivo model.

This study explored the structures of three polysaccharides from Bupleurum chinense DC. (BCPRs), and evaluated their antioxidant and anti-aging properties. The HPGPC and ion chromatography analyses revealed that the molecular weights of the BCPRs ranged from 12.05 to 21.20 kDa, and were primarily composed of rhamnose, arabinose, xylose, galactose, glucose and galacturonic acid. Methylation and NMR studies identified 10 PMAAs, establishing the various backbones of BCPRs 1-3. BCPR-3 demonstrated potent antioxidant activities, including DPPH, ABTS, hydroxy, and superoxide radicals scavenging in vitro. At concentrations between 125 and 500 µg/mL, BCPR-3 increased T-AOC, SOD and GSH-Px activities, while decreasing MDA levels in H2O2-induced SH-SY5Y cells. In addition, RNA-seq results indicated that BCPR-3 considerably downregulated the expression of 49 genes and upregulated five genes compared with the control group. KEGG analysis suggested that these differentially expressed genes (DEGs) were predominantly involved in the TNF and PI3K/Akt signaling pathways. Furthermore, in vivo experiment with Drosophila melanogaster showed that BCPR-3 could extend the average lifespan of flies. In conclusion, polysaccharides from B. chinense exhibited potential antioxidant and anti-aging activities, which could be developed as new ingredients to combat oxidative stress damage and slow the aging process.

Corrigendum: A minimally manipulated preservation and virus inactivation method for amnion/chorion.

[This corrects the article DOI: 10.3389/fbioe.2022.952498.].

CircPSMC3 Inhibits Colorectal Cancer Cell Growth and Migration by Controlling miR-31-5p/YAP/ß-Catenin.

INTRODUCTION: Colorectal cancer (CRC) is hackneyed cancer and a major lethiferous cancer. Circular RNAs (CircRNAs) have been discovered to own important roles in controlling CRC progression. CircPSMC3 is known to exhibit lower expression in diversified cancers. However, the regulatory function of CircPSMC3 in CRC keeps unclear. METHODS: The expression of CircPSMC3 and miR-31-5p was confirmed through RT-qPCR. The cell proliferation was measured through CCK-8 and EdU assays. The protein expression of genes was examined through a western blot. The cell invasion and migration were tested through Transwell and wound healing assays. The binding ability between CircPSMC3 and miR-31-5p was confirmed through the luciferase reporter assay. RESULTS: CircPSMC3 exhibited lower expression in CRC tissues and cell lines. Additionally, CircPSMC3 was revealed to suppress cell proliferation in CRC. Moreover, through Transwell and wound healing assays, CircPSMC3 was discovered to repress CRC cell invasion and migration. In CRC tissues, miR-31-5p expression was up-regulated and negatively correlated with CircPSMC3 expression. Further mechanism exploration experiments disclosed that CircPSMC3 is bound with miR-31-5p to modulate the YAP/ß-catenin axis in CRC. At last, through rescue assays, CircPSMC3 inhibited cell proliferation, invasion and migration through sponging miR-31-5p in CRC. CONCLUSION: Our work was the first time to probe the potential regulatory effects of CircPSMC3 in CRC, and these above results uncovered that CircPSMC3 inhibited CRC cell growth and migration through regulating miR-31-5p/YAP/ß-catenin. This discovery hinted that CircPSMC3 may serve as a useful therapeutic candidate for CRC.

Prognostic prediction of novel risk scores (AML-DRG and AML-HCT-CR) in acute myeloid leukemia patients with allogeneic hematopoietic stem cell transplantation.

We aimed to validate and prove the novel risk score models of acute myeloid leukemia (AML)-specific disease risk group (AML-DRG) and AML-Hematopoietic Cell Transplant-composite risk (AML-HCT-CR) in patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (AHCT). Among the 172 AML patients analysed, 48.3% (n = 83) were females. Median age was 31.5 years (range 14 to 62 years), two patients was more than 60 years old (1.2%). Median follow-up was 44 months (range 1 to 94 months). According to the AML-DRG model, 109, 49 and 14 patients were in low-, intermediate- and high-risk group, respectively. According to the AML-HCT-CR model, 108, 30, 20 and 14 patients were in low-, intermediate-, high- and very high-risk group, respectively. Our results showed that the AML-DRG and AML-HCT-CR models significantly predicted cumulative incidence of relapse (p < 0.001; p < 0.001). But AML-DRG model was not associated with NRM (p = 0.072). Univariate analysis showed that the AML-DRG model could better stratify AML patients into different risk groups compared to the AML-HCT-CR model. Multivariate analysis confirmed that prognostic impact of AML-DRG and AML-HCT-CR models on post-transplant OS was independent to age, sex, conditioning type, transplant modality, and stem cell source (p < 0.001; p < 0.001). AML-DRG and AML-HCT-CR models can be used to effectively predict post-transplant survival in patients with AML receiving AHCT. Compared to AML-HCT-CR score, the AML-DRG score allows better stratification and improved survival prediction of AML patients post-transplant.

KIF2A participates in the progression of hepatocellular carcinoma and angiogenesis by interacting with Notch1.

Kinesin family member 2A (KIF2A) serves a vital role in the development of hepatocellular carcinoma (HCC); however, the biological effect of KIF2A on the malignant progression of HCC remains unclear. Therefore, the present study was conducted to systematically determine the biological role of KIF2A in HCC and to better understand the molecular mechanism. The differences of KIF2A expression in HHL-5 normal human hepatocytes and the HCC cell lines Li-7, Huh7 and MHCC97 were assessed by reverse transcription-quantitative PCR and western blotting analysis. Moreover, viability, proliferation, migration and invasion of HCC cells were assessed by performing CCK-8, 5-ethynyl-2'-deoxyuridine staining, wound healing and Transwell assays. Additionally, the tube formation assay was performed to evaluate angiogenesis of HUVECs incubated with the conditioned media of HCC cells in vitro. The interaction between KIF2A and Notch1 was analyzed through co-immunoprecipitation assay. KIF2A was revealed to be highly expressed in HCC cells. KIF2A knockdown suppressed HCC cell proliferation, migration and invasion, and impaired in vitro angiogenesis. Furthermore, it was revealed that KIF2A interacted with Notch1 and positively regulated Notch1 expression. The suppressive effects of KIF2A knockdown on HCC cell proliferation, migration, invasion and in vitro angiogenesis were partially reversed by Notch1 overexpression. Overall, KIF2A may act as an oncogene in HCC via activation of the Notch1 signaling pathway.

Application of pedicled greater omentum flap tamponade combined with laparoscopic fenestration in diaphragmatic hepatic cyst.

OBJECTIVE: To investigate the efficacy and clinical application advantage of omental tamponade with vascular pedicle combined with Laparoscopic fenestration for the treatment of diaphragmatic hepatic cyst. METHODS: A total of 56 patients with diaphragmatic hepatic cysts underwent laparoscopic surgery in a single tertiary academic medical center from January 2010 to October 2020, including 21 patients (non-omental group) underwent laparoscopic fenestration of liver cysts, and 36 patients underwent laparoscopic liver cyst fenestration combined with vascular pedicle omentum tamponade (omental group). The general conditions and follow-up results of the two groups were compared and annalyzed. RESULTS: The operation time of the omental group was longer than that of the non-omental group (P = 1.358E-4). There was no significant difference in postoperative complications, postoperative laboratory values and hospital costs (P>0.05). The length of hospital stay in omental group was shorter than that in non-omental group (P = 0.034). In the omental group, recurrence occurred in 1 of 35 patients (4.65%) who were followeded up 12 months after surgery. In the non-omental group, of the 21 patients followed, 3 patients (14.28%) recurred 6 months after surgery, and 8 patients (38.10%) recurred 12 months after surgery. CONCLUSION: It is an effective method to prevent the recurrence of diaphragmatic hepatic cyst after laparoscopic fenestration by packing the cyst with vascularized omentum.

A minimally manipulated preservation and virus inactivation method for amnion/chorion.

Allogeneic amnion tissues have been widely used in tissue repair and regeneration, especially a remarkable trend of clinical uses in chronic wound repair. The virus inactivation procedures are necessary and required to be verified for the clinical use and approval of biological products. Cobalt-60 (Co-60) or electron-beam (e-beam) is the common procedure for virus and bacterial reduction, but the excessive dose of irradiation was reported to be harmful to biological products. Herein, we present a riboflavin (RB)-ultraviolet light (UV) method for virus inactivation of amnion and chorion tissues. We used the standard in vitro limiting dilution assay to test the viral reduction capacity of the RB-UV method on amnion or chorion tissues loaded with four types of model viruses. We found RB-UV was a very effective procedure for inactivating viruses of amnion and chorion tissues, which could be used as a complementary method to Co-60 irradiation. In addition, we also screened the washing solutions and drying methods for the retention of growth factors.

Selenium-containing polysaccharides isolated from Rosa laevigata Michx fruits exhibit excellent anti-oxidant and neuroprotective activity in vitro.

Selenium-containing polysaccharides have potential as an organic selenium dietary supplement, owing to their low toxicity, few side effects, and easy absorption attributes. In this study, we isolated two novel homogeneous selenium-containing polysaccharides from Rosa laevigata Michx fruits (Se-RLFPs). Results from primary structural analysis revealed that Se-RLFPs were α - pyranose, and were both composed of rhamnose, xylose, glucose with an average molecular weight of 24 and 16 KDa, respectively. Selenium contents in Se-RLFP-I and Se-RLFP-II were 16.49 µg/g and 21.61 µg/g, respectively. Results from analysis of antioxidant and neuroprotective activity of the polysaccharides revealed that Se-RLFPs had a radical scavenging effect. Specifically, they effectively protected SH-SY5Y cells from H2O2-induced damage by enhancing antioxidant enzyme activities (SOD), total antioxidant capacity (T-AOC) and suppressing malondialdehyde (MDA) levels. Western blots showed that the underlying mechanisms of action may be related to the Nrf2/HO-1 signaling pathway. Taken together, these results suggested that Se-RLFPs have potential as a pharmaceutical agent for treatment of neurodegenerative diseases (NDDs) or as a selenium-complementary ingredient in functional foods.

Differences in the properties of extracellular polymeric substances responsible for PAH degradation isolated from Mycobacterium gilvum SN12 grown on pyrene and benzo[a]pyrene.

This study aimed to evaluate the differences in the characteristics of extracellular polymeric substances (EPSs) secreted by Mycobacterium gilvum SN12 (M.g. SN12) cultured on pyrene (Pyr) and benzo[a]pyrene (BaP). A heating method was used to extract EPSs from M.g. SN12, and the composition, emulsifying activity, and morphology of EPS extracts were investigated. Results showed that EPS extracts varied significantly with Pyr or BaP addition to the bacterial cultures. The concentration of proteins and carbohydrates, the main components of the EPS extracts, first increased and then decreased, with an increase in the concentration of Pyr (0-120 mg L-1) and BaP (0-120 mg L-1). A similar trend was observed for the emulsifying activity of the EPS extracts. EPSs extracted from all cultures exhibited a compact structure with a smooth surface, except for EPSs extracted from BaP-grown M.g. SN12, which revealed a more fragile and softer surface. These findings suggest that Pyr and BaP had different influences on the properties of isolated EPSs, providing insights into the mechanism underlying polycyclic aromatic hydrocarbons (PAHs) biodegradation by some EPS-secreting bacteria. To the best of our knowledge, this is the first report on the texture profile of EPS samples extracted from M.g. SN12 grown on PAHs.

Influences of impervious surfaces on ecological risks and controlling strategies in rapidly urbanizing regions.

Reducing ecological risks is important for promoting regional sustainable development. However, studies on the influence of impervious surfaces on ecological risks and risk control strategies in regions undergoing rapid urbanization are limited. Therefore, this study aimed to demonstrate the spatial-temporal dynamics of regional ecological risks using Beijing as a case study to reveal the influence of impervious surfaces and explore the controlling strategies of risks. We first characterized the ecological risks in Beijing based on the ecosystem service values and mapped the risk levels and temporal variations in risks. We then identified the ecological risk increases caused by impervious surface expansion and built linear regression models for impervious surface coverage (ISC) and risk index. Finally, we formulated ecological risk control strategies for the strategy categories identified based on the ISC thresholds. The results show that the mountainous areas mainly exhibited low ecological risk levels, and the plain areas mainly showed high levels. The expansion of impervious surface was the main cause of the relatively large temporal increase in ecological risks from 2005 to 2015. Moreover, the strategies for ecological risk control can be divided into four categories based on the division of ISC, with 30%, 70%, and 90% as the thresholds. For risk control strategies, reducing ISC is the most important measure to reduce ecological risks for the category with an ISC range of 90%-100%, and increasing the area proportions of forests and water bodies is the most effective measure for the category with an ISC range of 0%-30%. For the other two categories, controlling the ISC and other strategies are required. Our study can increase the understanding of the influences of impervious surfaces on ecological risks in rapidly urbanizing regions and help inform the formulation of strategies for controlling the ecological risks in Beijing.

Sprayable methacrylic anhydride-modified gelatin hydrogel combined with bionic neutrophils nanoparticles for scar-free wound healing of diabetes mellitus.

Hard-to-healing or nonhealing diabetic wounds caused by hyperglycemia, bacterial infection and chronic inflammation are becoming a challenge globally. In this study, a novel hydrogel for diabetic wound healing composed of methacrylic anhydride-modified gelatin (GelMA) hydrogel and mimicking neutrophil nanoparticles was originally created. The prepared GelMA hydrogel has good sprayability and film-formation ability under blue light illumination (wavelength = 435-480 nm). Nanoparticles mimicking neutrophils belong to a double enzyme system that are encapsulated in ZIF-8 nanoparticles, which can consume glucose to produce HClO, ensuring a decrease in the glucose concentration of the wound and growth inhibition in bacteria. The hydrogel also has excellent biocompatibility, which can promote the growth and proliferation of fibroblasts. More importantly, the hydrogel can accelerate wound healing in type I diabetic rats owing to the downregulation of proinflammatory cytokines, and the wound with an area of 1 cm2 can be almost fully healed with no formation of the scar on the 21st day, as verified by histochemistry and immunohistochemistry. All these combinations indicate its potential in diabetic wound treatment.

The application of urban anthropogenic background to pollution evaluation and source identification of soil contaminants in Macau, China.

The anthropogenic background characterized by the accumulation characteristics of contaminants is recognized as an important evidence in pollution assessment and source identification in urban soil due to its less arbitrariness compared with the existing quality standards and the guidelines. A credible approach for pollution index calculation referring to anthropogenic background values (ABVs) combined with entropy weight method was developed. By the approach, the soil pollution degrees in Macau, China (one of the most densely populated region worldwide) were assessed based on the database of the heavy metals, Cd, Cu, Hg, Pb, and Zn, and high molecular weight polycyclic aromatic hydrocarbons (HMW PAHs) from 31 sites spatially distributed all over Macau. It was revealed that approximately half of the sites had no specific point source pollution. Mercury, benzo(a)anthracene (BaA), fluoranthene (FLT), and benzo(b)fluorantene (BbF), which had the highest weights were considered as the main contaminants. Macau Peninsula was identified as the critical polluted area. Then, the positive matrix factorization (PMF) coupled with ABVs as one of the data uncertainty inputs was used to identify the anthropogenic pollution sources of the contaminants. Three main anthropogenic sources with their contributions, including vehicle emissions (51.3%), use of hazard material (24.8%), and municipal or domestic waste (23.9%), could be well identified and quantified in the study area. The error estimation of the results showed that the variation of the contaminants in the derived factors were stable. The approaches which were in conformity with ABVs of soil contaminants are proved applicable in soil pollution assessment and source identification.

Coupling relation between urbanization and ecological risk of PAHs on coastal terrestrial ecosystem around the Bohai and Yellow Sea.

Urbanization, the ecological risks of polycyclic aromatic hydrocarbons (PAHs) to terrestrial ecosystems, and the complex relationship between them have drawn globally attention. In this paper, a comprehensive indicator system was calculated to illustrate the levels of urbanization in 20 coastal cities around the Bohai and Yellow Sea. The top three cities with high levels of urbanization were Tianjin > Qingdao > Dalian. The ecological risk of phenanthrene (Phe) was 52.0%; while the risk of fluoranthene (Flt) was 25.8%, and that of pyrene (Pyr), benzo[a]anthracene (Bap), fluorene (Flu), and naphthalene (Nap) were below 20% throughout the entire region. Risks were high in Dandong, Tianjin, Tangshan, Nantong, and Lianyungang and low in Weihai, Dongying, and Rizhao. The degree of coupling between urbanization and ecological risk of PAHs was above high (0.6) and more than 50% of the coordination degrees were slight unbalance [0.3, 0.5). Furthermore, redundancy analysis showed that the indicator aspects of industry, transportation, and population made great contribution to PAHs risk. Industry correlated to Low Molecular Weight (LMW) PAHs, while transportation correlated to High Molecular Weight (HMW) PAHs. To minimize risk, urbanization scale should be under acceptable level, or the structure of industry and transportation should be optimized.

Amidase as a versatile tool in amide-bond cleavage: From molecular features to biotechnological applications.

Amidases (EC 3. 5. 1. X) are versatile biocatalysts for synthesis of chiral carboxylic acids, α-amino acids and amides due to their hydrolytic and acyl transfer activity towards the C-N linkages. They have been extensively exploited and studied during the past years for their high specific activity and excellent enantioselectivity involved in various biotechnological applications in pharmaceutical and agrochemical industries. Additionally, they have attracted considerable attentions in biodegradation and bioremediation owing to environmental pressures. Motivated by industrial demands, crystallographic investigations and catalytic mechanisms of amidases based on structural biology have witnessed a dramatic promotion in the last two decades. The protein structures showed that different types of amidases have their typical stuctural elements, such as the conserved AS domains in signature amidases and the typical architecture of metal-associated active sites in acetamidase/formamidase family amidases. This review provides an overview of recent research advances in various amidases, with a focus on their structural basis of phylogenetics, substrate specificities and catalytic mechanisms as well as their biotechnological applications. As more crystal structures of amidases are determined, the structure/function relationships of these enzymes will also be further elucidated, which will facilitate molecular engineering and design of amidases to meet industrial requirements.

Structural features, selenization modification, antioxidant and anti-tumor effects of polysaccharides from alfalfa roots.

Hot water extraction and chromatographic purification methods were used to extract and purify two polysaccharides (RAPS-1 and RAPS-2) from the roots of alfalfa. Subsequently, RAPS-2 was modified using the HNO3/Na2SeO3 method to obtain Se-RAPS-2. The structural features, antioxidant and in vitro anti-tumor activities of the three polysaccharides were evaluated. The structural analysis revealed that RAPS-1 (Mw = 10.0 kDa) was composed of rhamnose, xylose, arabinose, galacturonic acid, mannose and glucose, whereas RAPS-2 (Mw = 15.8 kDa) consisted of rhamnose, xylose, galacturonic acid, mannose, glucose and galactose. RAPS-1 contained 1 â†’ 2, 1 â†’ 4, 1 â†’ 3, and 1 â†’ 6 or 1 â†’ glycosidic bonds; however, while RAPS-2 lacked 1 â†’ 4 glycosidic linkages. The molecular weight of Se-RAPS-2 was 11.0 kDa less than that of RAPS-2. The results of activities demonstrated that Se-RAPS-2 displayed superior antioxidant activity and inhibitory effect in HepG2 cells than RAPS-1 and RAPS-2.

Polydatin Exerts an Antitumor Effect Through Regulating the miR-382/PD-L1 Axis in Colorectal Cancer.

Background: Colorectal cancer (CRC) is considered as one of the most lethal malignancies worldwide. However, the effective therapies remain limited. Polydatin, a main effective component of the Chinese herb Polygonum cuspidatum, has multiple antitumor activities; however, whether Polydatin has anti-CRC activity is not fully understood. Materials and Methods: CRC cell proliferation and apoptosis were measured after treatment of Polydatin using Cell Counting Kit-8 assay, colony formation assay, and flow cytometer assay. The expression of miR-382 and programmed cell death ligand 1 (PD-L1) were determined in CRC cell lines by quantitative real-time polymerase chain reaction and Western blot, respectively. Furthermore, dual-luciferase reporter assay was conducted to determine the target of miR-382. Moreover, loss-of-functional experiments were used to identify the effect of Polydatin on miR-382. Finally, tumor xenograft experiments were conducted to determine the effect of Polydatin in vivo. Results: As a result, Polydatin effectively inhibited cell proliferation and promoted cell apoptosis in CRC cell lines. PD-L1 was confirmed as a direct target of miR-382. Furthermore, Polydatin could suppress the expression of PD-L1 by upregulating miR-382. Moreover, Polydatin inhibits proliferation and promotes apoptosis of CRC cells by regulating miR-382 and suppressing CRC tumor growth in vivo. Conclusion: Polydatin inhibits CRC cell proliferation and promotes apoptosis by regulating miR-382/PD-L1 axis. Polydatin could be a potential compound to synthesize novel antitumor drugs.

A high whey protein, vitamin D and E supplement preserves muscle mass, strength, and quality of life in sarcopenic older adults: A double-blind randomized controlled trial.

OBJECTIVE: Sarcopenia, an age-related decline of muscle mass, strength, and physical function, was associated with falls, frailty, and poor quality of life. The aim of the current study is to examine the effect of nutritional supplement containing whey protein, vitamin D and E on measures of sarcopenia. METHODS: A total of 60 sarcopenic older adult subjects participated in the current randomized, double-blind, placebo-controlled (iso-caloric control product) trial for 6 months. Muscle mass [Relative skeletal mass index (RSMI) measured by bioimpedance analysis (BIA)], muscle strength (handgrip strength), physical function (6-m gait speed, chair stand test, and timed-up-and-go test, TUG), quality of life (measured by Short-Form 36-Item Health Survey, SF-36), and blood biochemical indexes were measured before and after the 6-month intervention. RESULTS: Compared to placebo group, nutritional supplementation improves RSMI (mean difference: 0.18 kg/m2, 95%CI: 0.01-0.35, P = 0.040), handgrip strength (mean difference: 2.68 kg, 95%CI: 0.71-4.65, P = 0.009), SF-36 mental component summary (SF-36 MCS) (mean difference: 11.26, 95%CI: 3.86-18.65, P = 0.004), SF-36 physical component summary (SF-36 PCS) (mean difference: 20.21, 95%CI: 11.30-29.12, P < 0.001), serum IGF-1 (mean difference: 14.34 ng/mL, 95%CI: 2.06-26.73), IL-2 (mean difference: -575.32 pg/mL, 95%CI: -1116.94 âˆ¼ -33.70, P = 0.038), serum vitamin D3 (mean difference: 11.01 ng/mL, 95%CI: 6.44-15,58, P < 0.001), and serum vitamin E (mean difference: 4.17 ng/L, 95%CI: 1.89-6.45, P = 0.001). CONCLUSION: The current study demonstrated that the combined supplementation of whey protein, vitamin D and E can significantly improve RSMI, muscle strength, and anabolic markers such as IGF-I and IL-2 in older adults with sarcopenia. Further larger well-designed studies are warranted to evaluate whether long-term whey protein supplementation can blunt the declines of muscle function and mass in older adults with sarcopenia.

The neuroprotective and antioxidant profiles of selenium-containing polysaccharides from the fruit of Rosa laevigata.

Rosa laevigata fruit has been known as a functional foodstuff for a long time. Recently, increasing attention has been given to polysaccharides from R. laevigata fruit due to their numerous medicinal and nutritional properties. In this study, a rapid and effective approach for the extraction and separation of polysaccharides from the title fruit was developed using microwave-assisted aqueous two-phase extraction (MA-ATPE) with a PEG/ammonium sulfate system. After analysis of the response surface methodology (RSM) data based on a Box-Behnken design (BBD), a model was proposed and was found to predict an optimum yield value of 258.99 mg g-1 which is in good agreement with the experimental value (258.59 mg g-1). Two selenium (Se)-containing polysaccharides, Se-RLFP-1 and Se-RLFP-2, were isolated from R. laevigata fruit. Their chemical structures were elucidated by acid hydrolysis, weight-average molecular mass and Se-content analysis, along with UV, FT-IR, 1H and 13C NMR spectroscopy. As a result, Se-RLFP-1 was found to be mainly composed of mannose, glucose, galactose and xylose in a molar ratio of 1.4 : 7.9 : 1.0 : 1.5, while Se-RLFP-2 was composed of mannose, rhamnose, glucose, galactose and xylose (12.6 : 1.0 : 38.3 : 5.6 : 19.6). Furthermore, the antioxidant properties of the polysaccharides were investigated on the basis of FRAP, ABTS and DPPH radical scavenging assays. The results showed that the two polysaccharides had a noticeable effect on the radical scavenging of ABTS and DPPH, especially at high concentrations. In addition, the neuroprotective effect of Se-RLFP-1 and Se-RLFP-2 against oxidative stress induced by H2O2 in SH-SY5Y neuroblastoma cells was also investigated. In particular, Se-RLFP-1 exhibited obvious neuroprotective activity at a concentration of 100 µg mL-1.

Rapid diagnosis of sepsis with TaqMan-Based multiplex real-time PCR.

BACKGROUND: The survival rate of septic patients mainly depends on a rapid and reliable diagnosis. A rapid, broad range, specific and sensitive quantitative diagnostic test is the urgent need. Thus, we developed a TaqMan-Based Multiplex real-time PCR assays to identify bloodstream pathogens within a few hours. METHODS: Primers and TaqMan probes were designed to be complementary to conserved regions in the 16S rDNA gene of different kinds of bacteria. To evaluate accurately, sensitively, and specifically, the known bacteria samples (Standard strains, whole blood samples) are determined by TaqMan-Based Multiplex real-time PCR. In addition, 30 blood samples taken from patients with clinical symptoms of sepsis were tested by TaqMan-Based Multiplex real-time PCR and blood culture. RESULTS: The mean frequency of positive for Multiplex real-time PCR was 96% at a concentration of 100 CFU/mL, and it was 100% at a concentration greater than 1000 CFU/mL. All the known blood samples and Standard strains were detected positively by TaqMan-Based Multiplex PCR, no PCR products were detected when DNAs from other bacterium were used in the multiplex assay. Among the 30 patients with clinical symptoms of sepsis, 18 patients were confirmed positive by Multiplex real-time PCR and seven patients were confirmed positive by blood culture. CONCLUSION: TaqMan-Based Multiplex real-time PCR assay with highly sensitivity, specificity and broad detection range, is a rapid and accurate method in the detection of bacterial pathogens of sepsis and should have a promising usage in the diagnosis of sepsis.

miR-27b-3p suppresses cell proliferation, migration and invasion by targeting LIMK1 in colorectal cancer.

Accumulating evidence has indicated that microRNAs (miRNAs) can act as candidate tumor suppressors. However, the mechanism of miR-27b-3p in colorectal cancer (CRC) is still unknown. In this study, the relative expression levels of miR-27b-3p and LIMK1 were detected by quantitative real-time PCR (qRT-PCR) in CRC and adjacent normal tissues as well as in CRC cell lines. The proliferation, cell cycle and migratory and invasive abilities of SW620 cells were analyzed by CCK-8, flow cytometry, wound healing assays, and Transwell assays, respectively. The regulatory relationships between LIMK1 and miR-27b-3p were detected by qRT-PCR, luciferase reporter gene and Western blot assays. The results showed that the expression levels of miR-27b-3p were downregulated in CRC samples compared with those in adjacent nontumor tissues. Moreover, we found that miR-27b-3p suppressed CRC cell proliferation, cycle and migration and invasion. Additionally, there was an inverse correlation between LIMK1 expression and miR-27b-3p expression in CRC tissues. LIMK1 was identified as a direct target of miR-27b-3p. Our findings indicated that miR-27b-3p inhibited CRC cell proliferation, migration and invasion by targeting LIMK1.