Development and application of a cycleave dual-probe fluorescence quantitative PCR method for simultaneous detection of Mycoplasma gallisepticum ts-11 vaccine strain and non-ts-11 strains.

An attenuated vaccine against the Mycoplasma gallisepticum ts-11 strain has become an effective prevention and control method against MG infection. However, the ts-11 strain is usually difficult to distinguish from the non-ts-11 strain (including field isolates and other vaccine strains (F and 6/85)). Therefore, it is critical to establish a rapid and effective method to distinguish ts-11 strains from non-ts-11 strains. The gene sequences of the ts-11 strain (CP044225.1) and the non-ts-11 strain (including the wild-type (CP006916.3), 6/85 (CP044224.1), and F strains (NC_017503.1) were used to construct a conserved region containing a single point mutation in the potC gene in the ts-11 strain, after which a primer-probe combination method was designed. The primer-probe method was able to accurately and efficiently identify the ts-11 and non-ts-11 strains with minimum detection limits of 2.43 copies/µL and 1.65 copies/µL, respectively. Moreover, it could simultaneously distinguish the ts-11 strain from a non-ts-11 strain, and amplifications of avian influenza virus, infectious bronchitis virus, Newcastle disease virus, fowl adenovirus, infectious laryngotracheitis virus, infectious bursal disease virus, chicken anemia virus, Marek's disease virus, Mycoplasma synoviae, and Ornithobacter rhinotracheale were negative. The detection of clinical samples revealed that the established dual-probe fluorescence quantitative PCR method could be used to screen for mixed and single infections of the ts-11 strain and non-ts-11 strains effectively, with lower variation coefficients for intra- and interbatch repetition. The established cycleave dual-probe fluorescence quantitative PCR method showed good specificity, sensitivity, and repeatability and provides powerful technical support for the rapid and efficient differential diagnosis of the MG ts-11 strain from non-ts-11 strains.

Novel binary regulated silicon-carbon materials as high-performance anodes for lithium-ion batteries.

The massive volume dilation, unsteady solid electrolyte interphase, and weak conductivity about Si have failed to bring it to practical applications, although its potential capacity is up to 4200 mAh g-1. For solving these problems, novel binary regulated silicon-carbon materials (Si/BPC) were done by a sol-gel procedure combined with single carbonization. Analytical techniques were systematically utilized to examine the effects of element doping at several gradients on morphology, structure and electrochemical properties of composites, thus the optimal content was identified. Si/BPC preserves a discharge specific capacity of 1021.6 mAh g-1with a coulomb efficiency of 99.27% after 180 cycles at 1000 mA g-1, within the upgrade than single-doped and undoped. In rate test, it has a specific capacity of 1003.2 mAh g-1at a high current density of 5000 mA g-1, quickly back towards 2838.6 mAh g-1at 200 mA g-1. The inclusion of B and P elements is linked to the electrochemical characteristics. In the co-doped carbon layers, the synergistic impact of doping B and P accelerates the diffusion kinetics of lithium ions, boosts diffusion rate of Li+, offers low electrochemical impedance (45.75 Ω). This brings more defects to provide transport carriers and induces a substantial amount of electrochemically active sites, which fosters the storage of Li+, thus making silicon material electrochemically more active and potential.

Copolyester toughened poly(lactic acid) biodegradable material prepared by in situ formation of polyethylene glycol and citric acid.

Polylactic acid (PLA) is a high-modulus, high-strength bio-based thermoplastic polyester with good biodegradability, which is currently a promising environmentally friendly material. However, its inherent brittleness has hindered its widespread use. In this study, we reported a simple and non-toxic strategy for toughening PLA, using biodegradable materials such as polyethylene glycol (PEG) and citric acid (CA) as precursors. Through reactive melt blending with PLA, PEG and CA form PEGCA copolyesters in situ during blending. At the same time, CA can react with PLA and PEG, forming a copolyester structure at the interface of the two phases, improving the interfacial compatibility between PEG and PEGCA with PLA. Fourier transform infrared spectroscopy confirms this. Experimental results show that when the content of PEG/CA reaches 15% (PLA/PEG/CA-15%) in the blends, the impact strength of the blend was 4.47 kJ m-2, and the maximum elongation at break was as high as 360.1%, which were about 2 and 44 times higher than those of pure PLA, respectively. Moreover, the tensile strength was still maintained at the level of 70%. This work can expand the application of PLA in food packaging and medical supplies.

Estrogen inhibits colonic smooth muscle contractions by regulating BKß1 signaling.

The estrogen inhibits colonic smooth muscle contractions, which may lead to constipation. However, the mechanisms of inhibition are poorly understood. Therefore, the present study examined the effect of estrogen on rat colonic smooth muscle contractions and its potential association with the large-conductance Ca2+-activated K+ channels ß1 (BKß1) subunit. Twenty-four female Sprague Dawley rats were randomly assigned to 4 groups. After 2 weeks of intervention, the contraction activity of isolated colonic smooth muscle and the expression of BKß1 in colonic smooth muscle of rats were detected. Additionally, in order to investigate the effects of estrogen on BKß1 expression and calcium mobilization, in vitro experiments were conducted using rat and human colonic smooth muscle cells (SMCs). BKß1 shRNA was used to investigate whether calcium mobilization is affected by BKß1 in colonic SMCs. To explore the relationship between ERß and BKß1, serial deletions, site-directed mutagenesis, a dual-luciferase reporter assay, and chromatin immunoprecipitation assays were employed. In response to E2, colonic smooth muscle strips showed a decrease in tension, while IBTX exposure transiently increased tension. Furthermore, in these muscle tissues, BKß1 and α-SMA were found to be co-expressed. The E2 group showed significantly higher BKß1 expression. In cultured colonic SMCs, the expression of BKß1 was found to increase in the presence of E2 or DPN. E2 treatment reduced Ca2+ concentrations, while BKß1 shRNA treatment increased Ca2+ concentrations relative to the control. ERß-initiated BKß1 expression appears to occur via binding to the BKß1 promoter. These results indicated that E2 may upregulate BKß1 expression via ERß and inhibit colonic smooth muscle contraction through ERß by directly targeting BKß1.

Long noncoding RNA TFAP2A AS1 exerts promotive effects in non small cell lung cancer progression via controlling the microRNA 548a 3p/CDK4 axis.

In this study, we mainly focus on probing expression profile and detailed functions of long non coding RNA TFAP2A antisense RNA 1 (TFAP2A AS1) in non small cell lung cancer (NSCLC). Moreover, the mechanisms played by TFAP2A AS1 were unraveled comprehensively. Herein, a notable overexpressed TFAP2A AS1 in NSCLC was observed by TCGA and our own cohort. An increased TFAP2A AS1 level displayed a negative correlation with the overall survival of patients with NSCLC. Loss of function approaches illustrated that the absence of TFAP2A AS1 weakened NSCLC cell proliferation, colony formation, migration and invasion in vitro. Also, interference of TFAP2A AS1 caused in vivo tumor growth suppression. Mechanistically, TFAP2A AS1 could negative regulate microRNA 584 3p (miR 584 3p) as a competitive endogenous RNA. Furthermore, cyclin dependent kinase 4 (CDK4), a direct target of miR 584 3p, was positively controlled by TFAP2A AS1 in a miR 5184 3p dependent manner. Rescue function experiments corroborated that the anticancer activities of TFAP2A AS1 deficient on the oncogenicity of NSCLC cells were reversed by downregulating miR 584 3p or overexpressing CDK4. To sum up, TFAP2A AS1 exhibits cancer promoting roles in NSCLC through the adjustment of miR 584 3p/CDK4 axis.

Zinc finger protein 384 enhances colorectal cancer metastasis by upregulating MMP2.

Zinc finger proteins (ZNFs) serve key roles in tumor formation and progression; however, the functions and underlying mechanisms of dysregulated ZNF384 in colorectal cancer (CRC) are yet to be fully elucidated. Therefore, the present study initially aimed to investigate the expression levels of ZNF384 in CRC samples. Moreover, lentiviral ZNF384 overexpression and ZNF384 knockdown models were established in CRC cells. Transwell, wound healing and in vivo tail vein metastasis assays were carried out to evaluate the effects of ZNF384 on CRC cell metastasis. Furthermore, reverse transcription­quantitative PCR, western blotting, serial deletion, site­directed mutagenesis, dual­luciferase reporter and chromatin immunoprecipitation assays were conducted to investigate the potential underlying mechanisms. The results of the present study demonstrated that ZNF384 expression was markedly increased in CRC samples and this was associated with a poor prognosis. Notably, ZNF384 overexpression increased the levels of CRC cell invasion and migration, whereas ZNF384 knockdown inhibited CRC development. Moreover, the results of the present study demonstrated that ZNF384 mediated the expression of MMP2. MMP2 knockdown inhibited ZNF384­mediated CRC cell invasion and migration, whereas MMP2 overexpression ameliorated ZNF384 knockdown­induced inhibition of CRC progression. In addition, the results of the present study demonstrated that hypoxia­inducible factor 1α (HIF­1α) had the ability to bind to the ZNF384 promoter, thereby initiating ZNF384 expression. In human­derived CRC samples, the expression levels of ZNF384 were positively correlated with both MMP2 and HIF­1α expression. Collectively, these findings highlighted that ZNF384 may act as a prognostic marker and regulator of CRC metastasis.

Role of MRI­based radiomics in locally advanced rectal cancer (Review).

Colorectal cancer is the third most common type of cancer, with high morbidity and mortality rates. In particular, locally advanced rectal cancer (LARC) is difficult to treat and has a high recurrence rate. Neoadjuvant chemoradiotherapy (NCRT) is one of the standard treatment programs of LARC. If the response to treatment and prognosis in patients with LARC can be predicted, it will guide clinical decision­making. Radiomics is characterized by the extraction of high­dimensional quantitative features from medical imaging data, followed by data analysis and model construction, which can be used for tumor diagnosis, staging, prediction of treatment response and prognosis. In recent years, a number of studies have assessed the role of radiomics in NCRT for LARC. MRI­based radiomics provides valuable data and is expected to become an imaging biomarker for predicting treatment response and prognosis. The potential of radiomics to guide personalized medicine is widely recognized; however, current limitations and challenges prevent its application to clinical decision­making. The present review summarizes the applications, limitations and prospects of MRI­based radiomics in LARC.

Highly Efficient SO2 Sensing by Light-Assisted Ag/PANI/SnO2 at Room Temperature and the Sensing Mechanism.

Sulfur dioxide (SO2) is one of the most hazardous and common environmental pollutants. However, the development of room-temperature SO2 sensors is seriously lagging behind that of other toxic gas sensors due to their poor recovery properties. In this study, a light-assisted SO2 gas sensor based on polyaniline (PANI) and Ag nanoparticle-comodified tin dioxide nanostructures (Ag/PANI/SnO2) was developed and exhibited remarkable SO2 sensitivity and excellent recovery properties. The response of the Ag/PANI/SnO2 sensor (20.1) to 50 ppm SO2 under 365 nm ultraviolet (UV) light illumination at 20 °C was almost 10 times higher than that of the pure SnO2 sensor. Significantly, the UV-assisted Ag/PANI/SnO2 sensor had a rapid response time (110 s) and recovery time (100 s) to 50 ppm SO2, but in the absence of light, the sensors exhibited poor recovery performance or were even severely and irreversibly deactivated by SO2. The UV-assisted Ag/PANI/SnO2 sensor also exhibited excellent selectivity, superior reproducibility, and satisfactory long-term stability at room temperature. The increased charge carrier density, improved charge-transfer capability, and the higher active surface of the Ag/PANI/SnO2 sensor were revealed by electrochemical measurements and endowed with high SO2 sensitivity. Moreover, the light-induced formation of hot electrons in a high-energy state in Ag/PANI/SnO2 significantly facilitated the recovery of SO2 by the gas sensor.

MicroRNA-23b-3p targets non-SMC condensing I complex subunit G to promote proliferation and inhibit apoptosis of colorectal cancer cells via regulation of the PI3K/AKT signaling pathway.

Colorectal cancer (CRC) is one of the most common types of malignancy worldwide and has a poor prognosis. Non-SMC condensing I complex subunit G (NCAPG) has been reported to be upregulated in numerous types of malignant tumor. However, to the best of our knowledge, its clinicopathological and biological significance in CRC remain to be elucidated. The results of the present study revealed that NCAPG expression levels were upregulated in human CRC tissues and cell lines. The upregulated expression of NCAPG was positively associated with patient clinicopathological characteristics, such as differentiation and tumor size, and independently associated with poor survival. Consistent with the clinical observations, NCAPG was discovered to promote the proliferation and inhibit the apoptosis of CRC cells. Moreover, NCAPG-knockdown inhibited CRC cell proliferation by regulating the PI3K/AKT signaling pathway. Furthermore, NCAPG was identified as a potential target of microRNA (miR)-23b-3p, which was subsequently demonstrated to negatively regulate NCAPG expression. In conclusion, the findings of the current study indicated that the miR-23b-3p/NCAPG/PI3K/AKT signaling axis may play an important role in CRC carcinogenesis, and the status of the molecule may represent a promising prognostic marker for the disease.

Glutamate receptor, ionotropic, N­methyl D­aspartate­associated protein 1 promotes colorectal cancer cell proliferation and metastasis, and is negatively regulated by miR­296­3p.

N­methyl D­aspartate receptors (NMDARs) are closely associated with the development, growth and metastasis of cancer. Glutamate receptor, ionotropic, N­methyl D­aspartate­associated protein 1 (GRINA) is a member of the of the NMDAR family, and its aberrant expression is associated with gastric cancer. However, the role of GRINA in colorectal cancer (CRC) is not completely understood. In the present study, expression profiles of GRINA in several CRC databases were obtained and further verified using clinical CRC samples. The effects of GRINA overexpression on CRC progression both in vivo and in vitro were assessed. Briefly, cell proliferation was detected using MTT assay, and cell migration and invasion ability were evaluated by wound healing and Transwell assay. In addition, the molecular mechanism underlying the upregulated expression of GRINA in CRC was investigated. The regulatory association between GRINA and miR­296­3p was detected by luciferase assay, reverse transcription­quantitative PCR and western blotting. The results demonstrated that GRINA expression levels were significantly increased in tumor samples compared with those in healthy samples, and upregulated expression of GRINA was associated with a less favorable prognostic outcome in patients with CRC. GRINA overexpression significantly increased CRC cell proliferation, invasion and migration. Additionally, it was determined that GRINA was post­transcriptionally regulated by microRNA (miR)­296­3p. Together, the results of the present study suggested the potential importance of the miR­296­3p/GRINA axis and highlighted potential novel targets for the management of CRC.

Long noncoding RNA TFAP2A-AS1 exerts promotive effects in non-small cell lung cancer progression via controlling the microRNA-548a-3p/CDK4 axis as a competitive endogenous RNA.

In this study, we mainly focus on probing expression profile and detailed functions of long non-coding RNA TFAP2A antisense RNA 1 (TFAP2A-AS1) in non-small cell lung cancer (NSCLC). Moreover, the mechanisms played by TFAP2A-AS1 were unraveled comprehensively. Herein, a notable overexpressed TFAP2A-AS1 in NSCLC was observed by TCGA and our own cohort. An increased TFAP2A-AS1 level displayed a negative correlation with the overall survival of patients with NSCLC. Loss-of-function approaches illustrated that the absence of TFAP2A-AS1 weakened NSCLC cell proliferation, colony formation, migration and invasion in vitro. Also, interference of TFAP2A-AS1 caused in vivo tumor growth suppression. Mechanistically, TFAP2A-AS1 could negative regulate microRNA-584-3p (miR-584-3p) as a competitive endogenous RNA. Furthermore, cyclin-dependent kinase 4 (CDK4), a direct target of miR-584-3p, was positively controlled by TFAP2A-AS1 in a miR-5184-3p-dependent manner. Rescue function experiments corroborated that the anticancer activities of TFAP2A-AS1 deficient on the oncogenicity of NSCLC cells were reversed by downregulating miR-584-3p or overexpressing CDK4. To sum up, TFAP2A-AS1 exhibits cancer-promoting roles in NSCLC through the adjustment of miR-584-3p/CDK4 axis.

Cytochrome C Oxidase Assembly Factor 1 Homolog Predicts Poor Prognosis and Promotes Cell Proliferation in Colorectal Cancer by Regulating PI3K/AKT Signaling.

PURPOSE: Colorectal cancer (CRC) is one of the most common malignancies in the world. The prognosis of advanced CRC is still poor. The purpose of this study was to identify a gene expression profile associated with CRC that may contribute to the early diagnosis of CRC and improve patient prognosis. PATIENTS AND METHODS: Five pairs of CRC tissues and paracancerous tissues were used to identify causative genes using microarray assays. The prognostic value of Cytochrome C Oxidase Assembly Factor 1 Homolog (COA1) in CRC was assessed in 90 CRC patients. Loss-of-function assays, cell proliferation assays using Celigo and MTT, colony formation assays, a subcutaneous xenograft mouse model, and apoptosis assays were used to define the effects of downregulation of COA1 in CRC cells in vitro and in vivo. The underlying molecular mechanisms of COA1 in CRC were also investigated. RESULTS: The causative gene COA1 was identified through microarray analysis. COA1 expression in CRC was notably associated with pathologic differentiation, tumor size, and tumor depth. COA1 expression may act as an independent prognostic factor for overall survival of CRC. Knockdown of COA1 inhibited the proliferation of CRC cells in vitro and the tumorigenicity of CRC cells in vivo. Decreased COA1 expression induced apoptosis of CRC cells. Based on the microarray assay results comparing HCT116 cells transfected with lentivirus encoding anti-COA1 shRNA or negative control shRNA, ingenuity pathway analysis (IPA) revealed that the PI3K/AKT signaling pathway was significantly enriched. Moreover, CCND1, mTOR, AKT1, and MDM2 were identified as the downstream genes of COA1. CONCLUSION: These findings demonstrate that COA1 promotes CRC cell proliferation and inhibits apoptosis by regulating the PI3K/AKT signaling pathway. Our results implicate COA1 as a potential oncogene involved in tumor growth and progression of CRC.

Negative effects of long-term moderate salinity and short-term drought stress on the photosynthetic performance of Hybrid Pennisetum.

Plants are always suffering periods of soil water deficit and sustained soil salinity during their life cycle. Unraveling the mechanisms underpinning the responses of plants, especially the photosynthesis, to drought, salinity, and co-occurring stresses is critical for both the protection of natural vegetation and the stabilization of crop production. To better understand the downregulation of photosynthetic capability induced by soil salinity and drought, gas exchange parameters, leaf pigment contents, and chlorophyll (Chl) a fluorescence transients were analyzed in leaves of Hybrid Pennisetum. Our results showed that long-term moderate salinity, short-term drought, and the combination of these stressors decreased leaf pigment content by 11.4-31.5% and net photosynthetic rate (Pn) by 14.6-67.6% compared to those in untreated plants. The reduction of Pn in Hybrid Pennisetum under long-term salinity stress mainly occurred by stomatal limitation, whereas non-stomatal limitation played a dominant role under short-term drought stress. The changes in Chl a fluorescence kinetics (especially the appearance of the L-band and K-band) in both stress treatments showed that salinity and drought stress damaged the structural stability of photosystem II (PSII) and disturbed the equilibrium between the electrons at the acceptor and donor sides of PSII. Furthermore, although the negative effect of drought stress on leaf photosynthesis was much greater than that of salinity stress, moderate salt stress alleviated the negative effect of drought stress on the photosynthetic performance of Hybrid Pennisetum after long acclimation times.

Explaining the influence of the introduced base sites into alkali oxide modified CsX towards side-chain alkylation of toluene with methanol.

The effect of the introduced base sites in CsX with alkali oxide modification towards side-chain alkylation of toluene with methanol was systematically investigated in the present work. A series of alkali oxide modified CsX with a base properties (base strength and amount) gradient were prepared by impregnating CsX with alkali metal hydroxide. Here, CsX with different base strengths were selected as the parent zeolite, which is favorable to clarify the different types of base site in the modified CsX. The base properties of the samples were elucidated based on the CO2-TPD results as well as with characterization by XPS and FT-IR of adsorbed CO2. It was revealed that the base strength of the oxygen atoms with negative charge (O δ-) in the zeolite framework can be strengthened by the electron donating function of the alkali oxides in the supercage of CsX. Detailed analysis of the correlation between the reaction behaviors of side-chain alkylation reaction and the base properties of the indicated catalysts was carried out. It was found that the dehydrogenation of methanol to formaldehyde step was promoted by the increasing base properties of O δ- in the CsX framework and that was the main reason for the promotion of the whole side-chain alkylation reaction. However, it was also found that the stronger base properties of the alkali oxide modified CsX would enhance the formation of active hydrogen atoms, which aggravated the unwanted styrene conversion to ethylbenzene.

MicroRNA-130b promotes cell migration and invasion by inhibiting peroxisome proliferator-activated receptor-γ in human glioma.

Glioma is the most common and aggressive type of primary brain tumor. MicroRNA (miR)-130b functions as a tumor-associated miR. The dysregulation of miR-130b is involved in numerous biological characteristics and properties of certain types of cancer. The present study revealed the function and possible molecular mechanism of miR-130b in glioma cells, reporting that the level of miR-130b was markedly higher, increasing progressively as the histologic grade of the glioma increased, compared with the level in normal tissues. Additionally, the present study demonstrated that patients with high miR-130b expression exhibited a poor 3-year survival rate and miR-130b was an independent factor for predicting the prognosis of patients with glioma. The downregulation of miR-130b reduced invasion and migration in U373 and U87 cells. Furthermore, the downregulation of miR-130b increased peroxisome proliferator-activated receptor-γ (PPARγ) expression and inhibited epithelial-mesenchymal transition (EMT) in glioma cells. The present study identified PPARγ as a direct target of miR-130b in glioma in vitro. Furthermore, PPARγ knockdown was revealed to reduce the effect on EMT caused by the downregulation of miR-130b in U87 cells. The present study demonstrated that miR-130b promotes glioma proliferation, migration and invasion by suppressing PPARγ and subsequently inducing EMT.

Meta-analysis of CYP2E1 polymorphisms in liver carcinogenesis.

BACKGROUND: The CYP2E1 protein is a monooxygenase with certain polymorphisms linked to liver cancer. However, results from individual studies remain controversial. AIMS: To evaluate CYP2E1 polymorphisms in liver carcinogenesis through meta-analysis. METHODS: All studies about CYP2E1 polymorphisms and liver cancer were retrieved from seven major databases. Original data from each study were pooled and re-analyzed. RESULTS: Total of 16 articles with 4862 cases were selected, including 1820 cases of liver cancer and 3042 cases of controls. The c1 allelic frequency in the cases and controls was 83.3% and 85.3%, respectively. Five genetic variations were compared: dominant c1c2/c2c2 vs. c1/c1 (OR=0.987 (0.853, 1.141)), homozygous c2c2 vs. c1c1 (OR=0.767 (0.526, 1.119)), heterozygous c1c2 vs. c1c1 (OR=1.005 (0.854, 1.182)), recessive c2c2 vs. c1c2/c2c2 (OR=0.771 (0.530, 1.122)), and different alleles c2 vs. c1 (OR=0.947 (0.828, 1.082)). Pooled data were further analyzed based on ethnicity, control sources, and HWE (Hardy-Weinberg equilibrium). These results from stratified groups were similar to that of nonstratified groups. CONCLUSIONS: Our meta-analysis results suggest that there is no evidence for a major role of CYP2E1 polymorphism in liver carcinogenesis, but do not rule out the possibility in certain cases.

The Protective Effects of Trypsin Inhibitor on Hepatic Ischemia-Reperfusion Injury and Liver Graft Survival.

The aim of this study was to explore the protective effects of ulinastatin (urinary trypsin inhibitor, UTI) on liver ischemia-reperfusion injury (IRI) and graft survival. We employed mouse liver cold IRI and orthotopic liver transplantation (OLTx) models. UTI was added to lactated Ringer's (LR) solution for liver perfusion and preservation in vitro or combined with UTI injection intraperitoneally to the liver graft recipient. Our results indicated that UTI supplementation protected the liver from cold IRI in a dose-dependent manner and prolonged liver graft survival from extended cold preserved liver donors significantly. The underlying mechanism of UTI on liver IRI may be mediated by inhibition of proinflammatory cytokine release, increasing the expression of the antiapoptotic gene Bcl-2 and decreasing the expression of the proapoptosis genes of Caspase-3 and Bax, and further protects hepatocytes from apoptotic death and improves liver function.