The impact of LncRNA-SOX2-OT/let-7c-3p/SKP2 Axis on head and neck squamous cell carcinoma progression: Insights from bioinformatics analysis and experimental validation.

BACKGROUND: LncRNA SRY-box transcription factor 2 overlapping transcript (SOX2-OT) is linked to multiple cancers, but its specific role and mechanism in head and neck squamous cell carcinoma (HNSCC) remain poorly understood. METHODS: We harnessed clinical data and HNSCC transcriptome profiles from UCSC Xena, TCGA, and GEO databases. Employing various algorithms, we assessed the correlation between SOX2-OT expression and the HNSCC immune microenvironment. Differential expression analysis identified immune-enriched miRNAs (DEmiRNAs) and mRNAs (DEmRNAs). Utilizing miRanda, miRWalk, and Cytoscape, we constructed a ceRNA network encompassing SOX2-OT, DEmiRNAs, and DEmRNAs. A Sankey diagram visualized pivotal SOX2-OT-miRNA-mRNA-pathways. Functional assays validated SOX2-OT silencing effects in HNSCC cells. Luciferase reporter assays verified SOX2-OT/let-7c-3p/SKP2 relationships. Additionally, a xenograft mouse model revealed SOX2-OT's impact on xenograft growth and lung metastasis. RESULTS: SOX2-OT expression demonstrated a predominantly positive correlation with B lineage and VTCN1, while manifesting a negative correlation with Neutrophil and CD47 in HNSCC tissues. We discerned a ceRNA network comprising 65 DEmiRNAs and 116 DEmRNAs, while a protein-protein interaction (PPI) network revealed 97 protein nodes among DEmRNAs. Notably, the Sankey diagram spotlighted six key DEmRNAs intricately linked to the SOX2-OT-regulated DEmiRNAs immune-related pathway. Experimental assays established that SOX2-OT silencing exerted inhibitory effects on cell proliferation, migration, tumor growth, and lung metastasis within HNSCC cells, both in vitro and in vivo. We identified let-7c-3p as a target miRNA of SOX2-OT and SKP2 as a target mRNA of let-7c-3p. CONCLUSIONS: Our study establishes the critical SOX2-OT/let-7c-3p/SKP2 axis as a pivotal regulator of HNSCC tumorigenesis and metastasis.

Effects of diflubenzuron on shrimp (Neocaridina palmata) in freshwater systems dominated by submerged plant (Ceratophyllum demersum).

Diflubenzuron (DFB) is a benzoylbenzourea insect growth regulator widely used in agriculture, horticulture, and vector control. Therefore, it can easily pollute water bodies and cause harm to aquatic life and ecosystems. To evaluate the impact of DFB on atyid shrimp Neocaridina palmate, the insecticide was applied, at 0, 0.74, 2.222, 6.667, 20, and 60 µg L-1, to indoor systems dominated by submerged plant Ceratophyllum demersum. The highest no observed effect concentration and the lowest observed effect concentration was determined to be 0.167 and 0.536 µg L-1, respectively, as it was counted with either activity or immune-reactive content of chitobiase. Subcellular indices were more sensitive, with a lowest observed effect concentration below 0.107 µg L-1. Principal response curves (PRC) and principal component analysis (PCA) showed that DFB reduced the biomass of C. demersum and the content of chlorophyll-a and phycocyanin in the media. The biomass of periphyton were promoted at the high concentrations. According to the PRC and PCA, DFB reduced the bacterial population related to photoautotrophy, sulphur reduction, and sulphur oxidation and it promoted those related to photoheterotrophy, nitrate reduction, nitrate denitrification, and nitrogen fixation. Besides, DFB reduced fungi related to denitrification. PRC and PCA showed that DFB had a negative impact on pH and dissolved oxygen levels and a positive impact on NH4-N, NO2-N, PO4-P, and conductivity, suggesting the deterioration in quality of water. This study provided useful information for understanding the ecotoxicological effects of DFB at population and community levels.

Inhibiting Intestinal Krüppel-Like Factor 5 Impairs the Beneficial Role of Renal Denervation in Gut Microbiota in Rats with Heart Failure.

Krüppel-like factor 5 (KLF5) is critical in maintaining intestinal barrier function, and renal denervation (RDN) mitigates gut microbiota aberrations in rats with heart failure (HF). It is unclear whether intestinal KLF5 can be regulated by RDN and whether inhibiting intestinal KLF5 weakens the beneficial role of RDN on gut microbiota. Sprague-Dawley rats were distributed into a CG (sham transverse aortic constriction [TAC] and sham RDN), HF (induced by TAC), or RDN (underwent RDN after TAC) group or a CG.M, HF.M, or RDN.M group, which included the administration of the KLF5 inhibitor to the CG, HF, or RDN group, respectively. Transmission electron microscopy, mRNA, and protein expression of KLF5 and desmoglein 2 (DSG2) in jejunum and sequencing of the 16S rRNA gene in fecal samples were evaluated. KLF5 expression was lower in the RDN group than in the HF group (P < 0.001). The microvillus length, density, length-to-width ratio, and DSG2 expression were lower in the RDN.M group than in the RDN group, and the same trend was observed between the HF.M and HF groups (all P < 0.05). The gut bacterial community structure was altered after administration of a KLF5 inhibitor. The abundances of Proteobacteria, Gammaproteobacteria, Sutterella, and Prevotellaceae were higher, and the abundance of Firmicutes was lower in the RDN.M group than in the RDN group (all P < 0.05). These findings indicated that RDN suppressed intestinal KLF5 expression, and inhibiting intestinal KLF5 expression exacerbated the gut microbiota by impairing the intestinal barrier function in HF rats following RDN, which weakened the beneficial role of RDN on gut microbiota. IMPORTANCE Krüppel-like factor 5 (KLF5) is critical for the maintenance of intestinal barrier function. It is unclear whether intestinal KLF5 expression can be affected by renal denervation (RDN) in heart failure (HF) and whether inhibiting intestinal KLF5 expression exacerbates the gut microbiome and weakens the role of RDN in mitigating gut microbiome aberrations in HF rats after RDN. We demonstrated that RDN significantly suppressed intestinal KLF5 expression and that inhibiting intestinal expression of KLF5 exacerbated the gut microbiota and weakened the role of RDN in mitigating microbiota aberrations by impairing intestinal barrier function, resulting in an increase in bacteria harmful to cardiac function and a decrease in beneficial bacteria in HF rats following RDN. This study highlighted the important roles of intestinal KLF5 in modulating gut microbiota in HF and suggested that the influence of RDN on intestinal KLF5 was another possible role of RDN in HF besides downregulating the sympathetic nerve.

Achieving the low interfacial tension by balancing crystallization and film-forming ability of the cathode interlayer for organic solar cells.

A series of molecules with imide units bridged by the core of thiophene-based groups, namely N-dimethylaminopropyl-4-thiophene-1,8- naphthalimide (NT), bis(N-dimethylaminopropyl)-4-thiophene-1,8-naphthalimide (NTN), and bis(N-dimethylaminopropyl)-4-bithiophene-1,8-naphthalimide (N2TN), have been reported as cathode interfacial materials (CIMs) to realize low interfacial tension with the blend in organic solar cells (OSCs). We evaluated the Ohmic contact between the active layer and these cathode interlayers basedon various characterizations, which is of great significance for further understanding these imide-based interlayers. It turned out that the homogeneous and continuous NTN interlayer as a CIM balanced the factors of crystallization and film-forming property, and broke through the limitation of poor conductivity and high aggregation in our previous work. Moreover, compared with NT and N2TN, the NTN interlayer achieve a combination of good solubility in methanol, efficient electron mobility, and aligned work function. These advantages of NTN are conducive to the realization of high-efficient interfacial electron collection and transfer, thus improving the short-circuit current density (JSC) and filling factor (FF) of devices. Therefore, the binary OSCs (PM6:Y6) based on NTN engineered aluminium-cathode with excellent stability demonstrate a maximum power conversion efficiency (PCE) of 16.56 %, which is higher than NT (PCE = 1.34 %) and N2TN (PCE = 13.90 %). The enhanced performance is ascribed to the improvement of JSC and FF, which is originated from the outstanding conductivity and high-quality interface of NTN. Surprisingly, the PM6:Y6-based semitransparent device with NTN obtain a PCE of 13.43 % with an average visible transmittance of 17.79 %, which is better than traditional PDINO. This study highlights a potential strategy for enhancing the performance of OSCs by the interface engineering via decreasing the interfacial intension.

Effects of tebufenpyrad on freshwater systems dominated by Neocaridina palmata, Physa fontinalis, and Ceratophyllum demersum.

Tebufenpyrad are widely used for control leaf mites in orchard and may enter freshwater systems through runoff, spray drift, and so on. Few papers have reported the side effect of the pesticide on population dynamics of aquatic taxa such as shrimps, gastropods, macrophytes, phytoplankton, and bacteria. Here, we tested the effect of a single application of tebufenpyrad on Neocaridina palmata, Physa fontinalis, Ceratophyllum demersum, Simocephalus vetulus, Dolerocypris sinensis, and so on, by indoor systems. The TWA (Time-weighted average)-based highest no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for Neocaridina palmata, which were counted by the wet weight, were 0.67 and 2.33 µg/L, respectively, and the dose-related effect lasted 21 d. According to our study, chitobiase could be used to quantify the effects of the pesticide on shrimp despite the interference from P. fontinalis, which was finally corrected by employing of antibodies. The NOEC and LOEC were thus determined to be 1.41 and ≤ 5.64 µg/L, respectively, which were higher than the values that was counted by the wet weight. Principal component analysis (PCA) and principal response curve (PRC) investigation showed that the pesticide suppressed population of C. demersum, and phytoplankton, while the Physa fontinalis, S. vetulus, and D. sinensis were stimulated by the pesticide. Illumina MiSeq was used to determine the alteration in bacterial community within the systems. The results of PRC and PCA analyses showed that tebufenpyrad induced flora of nitrate reducing, nitrate denitrifying, thiosulfate oxidation, ureolysis, and methanol oxidation, while it suppressed flora of cellulolysis. Tebufenpyrad was found to have a negative effect on water quality indicators such as pH, DO, NO3-, NO2-, and SO42-, and a positive effect on PO43-, NH4+, and EC. This suggested that the tebufenpyrad led to water quality deterioration.

Spermidine Affects Cardiac Function in Heart Failure Mice by Influencing the Gut Microbiota and Cardiac Galectin-3.

Spermidine, which can be synthesized by the gut microbiota, can prevent cardiac hypertrophy and delay the progression to heart failure (HF). However, it is not clear whether the effect of spermidine on cardiac function is mediated by modulating the gut microbiota when HF occurs. Female HF Kunming mice induced by transverse aortic constriction were administered spermidine (HF+S group) or its antagonist (HF+SR group). Echocardiography, messenger ribonucleic acid (RNA) and protein expression of galectin-3 in the heart, cardiomyocyte apoptosis assays and gut microbiota analysis were detected. Left ventricular end-diastolic volume and diameter (LVVd and LVDd), and left ventricular end-systolic volume and diameter in the HF+SR group were significantly enlarged compared with those in the HF group (all P < 0.05). The HF+S group had a smaller LVDd and LVVd than the HF+SR group (5.01 ± 0.67 vs. 6.13 ± 0.45 mm, P = 0.033; 121.44 ± 38.74 vs. 189.94 ± 31.42 µL, P = 0.033). The messenger RNA and protein expression of galectin-3 and the number of apoptotic cardiomyocytes increased significantly in the HF+SR group compared to the HF group. Gut microbiota analysis showed that spermidine antagonists reduced the Firmicutes/Bacteroidetes ratio and changed the microbial community richness and diversity. In conclusion, spermidine can improve cardiac function in HF, and the regulation of gut microbiota and cardiac fibrosis may be a factor in the effect of spermidine on the improvement of cardiac function.

Renal Denervation Mitigated Fecal Microbiota Aberrations in Rats with Chronic Heart Failure.

BACKGROUND: Changes in the composition and diversity of gut microbiota, which can be altered by autonomic nerve activity, contribute to the development of heart failure (HF). Renal denervation (RDN) can improve cardiac function by reducing sympathetic nerve activity. However, whether the beneficial role of RDN on HF is related to gut microbiota is unknown. METHODS: Thirty rats were assigned to a control, HF (with induced transverse aortic constriction (TAC)), RDN (with RDN induced 10 weeks after TAC), Nog (HF rats with Nogo-P4-administered 8 weeks after RDN), and NEP (HF rats with NEP1-40-administered 8 weeks after RDN) group. Then, 16SrRNA amplicon sequencing and analyses of fecal samples were performed. RESULTS: Beta diversity analyses revealed that compared to the HF group, the RDN, Nog, and NEP groups clustered closer to the control group. The Firmicutes/Bacteroidetes ratio was reduced in the HF group (1.59) compared with the control group (3.21) and was significantly decreased compared to the Nog (7.19), RDN (6.20), and NEP (4.42) groups. At the genus level, the HF group showed decreased abundances of Lactobacillus and Alistipes and increased abundances of Bacteroides and Clostridium compared with the control group. The abundances of Lactobacillus and Alistipes were increased, and those of Bacteroides and Clostridium were decreased in the RDN, Nog, and NEP groups compared to the HF group. However, no differences were observed between the three groups that underwent RDN. The microbial function showed the same tendency. CONCLUSIONS: RDN reversed the abnormal changes in the gut microbiome in HF rats. Inhibition of reinnervation after RDN did not affect intestinal bacteria.

The influence of inhibiting renal neural regeneration on the efficacy of renal denervation to chronic heart failure.

AIMS: Some studies support the occurrence of nerve regeneration in renal arteries after renal denervation (RDN). But it is unclear whether inhibiting reinnervation after RDN is beneficial to enhancing the effect of RDN on chronic heart failure (CHF). METHODS AND RESULTS: Chronic heart failure Sprague Dawley rats induced by transverse aortic constriction were administered with the analogue of Nogo-B (Nogo group) or its antagonist (NEP group) respectively after RDN. Echocardiography, messenger RNA, and protein expression of calcitonin gene-related peptide (CGRP) in renal artery and nerves surrounding renal artery were detected. Relative protein expression of CGRP was significantly decreased in the Nog group compared with the RDN group (0.64 ± 0.51 vs. 1.68 ± 1.07, P = 0.048). The number of nerves surrounding renal artery was higher in the NEP group than in the Nog group. Left ventricular end-systolic volume and diameter (LVVs and LVDs) were greatly decreased, and left ventricular ejection fraction (LVEF) and fractional shortening (FS) increased significantly in the RDN, Nog and NEP groups when compared with the HF group (all P < 0.05). No significant differences were observed in left ventricular end-diastolic volume and diameter; LVDs; LVVs; FS; LVEF; and the levels of plasma renin, noradrenaline, and N-terminal pro-B-type natriuretic peptide among three groups: the RDN, Nog, and NEP groups. CONCLUSIONS: Reinnervation of renal artery occurred in CHF rats after RDN, which had no effect on therapeutic role of RDN in CHF, and inhibiting this neural regeneration had no clinical significance and did not affect the efficacy of RDN to CHF.

Direct and indirect effects of fenoxycarb on freshwater systems dominated by Neocaridina palmata (Decapoda: Atyidae) and macrophyte Ceratophyllum demersum.

Few studies have been conducted with regard to the effects of insecticides on population dynamics of shrimps and associated groups such as macrophytes, phytoplankton, microorganisms etc. In the present study, effects of a single application of fenoxycarb were tested using indoor freshwater systems dominated by Neocaridina palmata and Ceratophyllum demersum (Dicotyledons: Ceratophyllales). The no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for the N. palmata, as scaled by liberated chitobiase, were 6.48 µg/L and 27.76 µg/L, and the dose-related effect lasted for 14 days. Results of principal components analysis (PCA) and that of principal response curves (PRC) method showed that the biomass of C. demersum and concentrations of chlorophyll-a were suppressed, while the concentrations of phycocyanin were promoted. Illumina high-throughput sequencing was adopted to determine the diversity of bacteria and fungi in the media. Result of PCA and PRC showed that the fenoxycarb promoted photosynthetic bacteria (e.g. Cyanobacteria and Rhodobacterales) and suppressed groups involved in nitrogen and sulfur the transformation (e.g. Flavobacterium, hgcI_clade, Cystobasidium, Rhodotorula and Rhizobiales). Promotion in pathogen such as Pseudomonas and Cercozoa and suppression in beneficial taxa such as Novosphingobium and Rhodotorula were also sighted. Result of study suggested a water quality deterioration due to fenoxycarb applications.

Flexible Self-Repairing Materials for Wearable Sensing Applications: Elastomers and Hydrogels.

Flexible pressure and strain sensors have great potential for applications in wearable and implantable devices, soft robots, and artificial skin. The introduction of self-healing performance has made a positive contribution to the lifetime and stability of flexible sensors. At present, many self-healing flexible sensors with high sensitivity have been developed to detect the signal of organism activity. The sensitivity, reliability, and stability of self-healing flexible sensors depend on the conductive network and mechanical properties of flexible materials. This review focuses on the latest research progress of self-healing flexible sensors. First, various repair mechanisms of self-healing flexible materials are reviewed because these mechanisms contribute to the development of self-healing flexible materials. Then, self-healing elastomer flexible sensor and self-healing hydrogel flexible sensor are introduced and discussed respectively. The research status and problems to be solved of these two types of flexible sensors are discussed in detail. Finally, this rapidly developing and promising field of self-healing flexible sensors and devices is prospected.

Cascading effects caused by fenoxycarb in freshwater systems dominated by Daphnia carinata and Dolerocypris sinensis.

To evaluate the aquatic hazards of the insect juvenile hormone analogue fenoxycarb, a single application (0, 48.8, 156.3, 500, 1600, and 5120 µg/L) of it was done in indoor freshwater systems dominated by Daphnia carinata (daphnid) and Dolerocypris sinensis (ostracoda). The responses of zooplankton (counted by abundance and the activity and immuno-reactive content of free N-Acetyl-ß-D-glucosaminidase (NAGase)), phytoplankton (counted by chlorophyll and phycocyanin), planktonic bacteria and fungi, and some water quality parameters were investigated in a period of 35 d. Results of the study showed that the ostracoda was more sensitive than daphnid, with time-weighted average (TWA)-based no observed effect concentrations (NOECs) to be 8.45 and 12.66 µg/L in systems without humic acid addition (HA-) and to be 6.37 and 9.54 µg/L in systems with humic acid addition (HA+). The duration of treatment-related effects in the ostracoda population was longer than the daphnid population (21 vs. 14 days). Besides, the data analysis indicated that the toxicity of fenoxycarb was significantly enhanced in the HA+ systems. Owing to the reduced grazing pressure, the concentrations of chlorophyll and phycocyanin increased in the two highest treatments. The increase in photosynthesis along with a reduced animal excretion led to an increase in pH and a decrease in nutrient contents. These changes seemed to have an effect on the microbial communities. For example, the abundances of some opportunistic pathogens of aquatic animals (e.g. Aeromonas and Cladosporium) and organic-pollutant-degrading microorganisms (e.g. Ancylobacter and Azospirillum) increased significantly in microbial communities, but the abundances of Pedobacter, Candidatus Planktoluna, and Rhodobacter (photosynthetic bacteria) markedly decreased. This study provides useful information to understand the ecotoxicological impacts of fenoxycarb at the population and community levels while integrating the effects of HA on toxicity.

Molecular cloning and characterization of genes encoding chitinase and N-acetyl-ß-glucosaminidase in Dolerocypris sinensis.

Chitinases and N-acetyl-ß-glucosaminidase (NAG) are important in molting and growth of crustaceans. In ostracods, the genes encoding these enzymes have not been characterized. The aim of the present study was to clone the genes encoding chitinase (DsChi) and NAG (DsNAG) from the ostracod, Dolerocypris sinensis, elucidate the phylogenetic relationships between the cloned genes and known chitinolytic enzymes, and determine the expression patterns of these genes at different stages of growth in the presence of an environmental pollutant. The genes were amplified from the genomic DNA of the organism using polymerase chain reaction (PCR). The products from PCR were cloned and characterized with bioinformatics tools, and their expression patterns at different growth stages were determined using real-time quantitative PCR (qRT-PCR). Nine and five introns were identified in DsChi and DsNAG genes, respectively. When compared with protein sequences available in GenBank, chitinase from D. sinensis was most closely related to that of Macrobrachium nipponense (61 % homology). The NAG of D. sinensis was most closely related to that of Limulus polyphemus (55.6 % homology). Based on phylogenetic analysis of known chitinases from crustaceans and insects, the D. sinensis chitinase tightly clustered in the same branch with chitinases from species within the Malacostraca class. In contrast, NAG of D. sinensis was clustered with NAG from F. candida.The level of expression of DsChi mRNA was significantly higher than that of DsNAG throughout the period of growth (p < 0.05). Treatment of D. sinensis cells with fenoxycarb significantly downregulated the expressions of DsChi and DsNAG throughout the period of growth (p < 0.05). These results show that the protein products of DsChi and DsNAG possess remarkable biochemical properties characteristic of a chitinase and NAG, respectively.

Predictive value of serum myostatin for the severity and clinical outcome of heart failure.

BACKGROUND: The exact relationship between serum myostatin and the severity and prognosis of chronic heart failure (CHF) is unclear. In this study, we investigated the association between serum myostatin and the severity and prognosis in patients with CHF. METHODS: Two hundred and eighty-eight CHF patients and 62 healthy controls were studied. Cardiac ultrasound and serum myostatin, N-terminal pro-B-type natriuretic peptide (NT-proBNP) and other parameters were detected. CHF patients were divided into 3 groups according to tertiles of NT-proBNP or myostatin levels respectively. RESULTS: Serum myostatin levels were higher in CHF patients than in controls. New York Heart Association (NYHA) class IV patients had the highest levels of serum myostatin among the four NYHA classes. Compared with the low tertile NT-proBNP group, serum myostatin levels were significantly higher in the moderate and high tertile groups (15.47 ±â€¯4.25 vs. 14.18 ±â€¯3.69 ng/mL, p = .026; 16.28 ±â€¯5.34 vs. 14.18 ±â€¯3.69 ng/mL, p = .002). During 51-months follow-up, of 173 patients there were 36 deaths. Compared to survivors, nonsurvivors had significantly higher serum myostatin (18.11 ±â€¯4.52 vs. 14.85 ±â€¯5.11 ng/mL, p < .01). Patients in the high tertile myostatin group had lower survival rate (73.95% vs. 93.75%; p < .05) and larger number of CHF rehospitalization than those in the low tertile group. Cox regression analysis showed that serum myostatin was an independent predictor of mortality. CONCLUSIONS: Serum myostatin levels can reflect the severity of CHF and be a predictor of adverse prognosis in CHF patients.

Serum Peroxisome Proliferator-activated Receptor Gamma Coactivator-1α Related to Myocardial Energy Expenditure in Patients With Chronic Heart Failure.

BACKGROUND: Peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) plays key roles in controlling cardiac metabolism and function. Myocardial energy expenditure (MEE) can reflect myocardial energy metabolism and cardiac function. Whether the variation of PGC-1α can influence MEE levels in chronic heart failure (CHF) is unclear. Therefore, we investigated the relationship between PGC-1α and MEE. MATERIAL AND METHODS: We studied 219 patients with CHF and 66 healthy controls. MEE was measured according to echocardiographic parameters. Serum PGC-1α, N-terminal pro-B-type natriuretic peptide and other parameters were detected. Patients with CHF were divided into different groups according to the left ventricular ejection fraction (LVEF) and the tertile range of MEE. RESULTS: Serum PGC-1α was lower in the MEE 2 and 3 groups compared with controls (both P < 0.05). Patients in the MEE 2 (1.73 ± 0.83 versus 2.16 ± 0.82 ng/mL, P = 0.001) and 3 groups (1.65 ± 0.73 versus 2.16 ± 0.82 ng/mL, P < 0.001) possessed lower levels of PGC-1α than those in the MEE 1 group. Compared with high LVEF, patients with low LVEF had higher MEE (median, 167 versus 73 cal/minute, P < 0.05) and lower PGC-1α (1.71 ± 0.65 versus 1.95 ± 0.91 ng/mL, P = 0.032). Multivariate logistic regression analysis showed that MEE (OR = 0.517, 95% CI = 0.267-0.998, P = 0.049) and creatinine (OR = 2.704, 95% CI = 1.144-6.391, P = 0.023) were independently associated with increased PGC-1α. CONCLUSIONS: Serum PGC-1α was related to MEE and LVEF in patients with CHF and can reflect the degree of MEE and the systolic function of the left ventricle.

Relationship between the maximum carotid plaque area and the severity of coronary atherosclerosis.

BACKGROUND: Carotid artery plaque is associated with coronary artery disease (CAD). Besides the presence of plaque, plaque characteristics is also related to the severity of CAD. So the characteristic difference of carotid plaque may affect this assessment role. However, it is unclear whether the maximum carotid plaque area can reflect the extents and severity of CAD. METHODS: We enrolled 388 consecutive CAD patients and 45 controls, and 204 patients were studied after excluding 184 patients without carotid plaque or coronary angiogram. Carotid intima media thickness and carotid plaque were measured by carotid ultrasound. Coronary angiography was applied and Gensini score was calculated. Blood lipid and other parameters were also detected. RESULTS: The total and right maximum carotid plaque area were greater in high Gensini Score group than those in low and moderate score groups (both P<0.05) and multivessel disease patients possessed the highest maximum plaque area. The total and right maximum carotid plaque area were also higher in collateral circulation group than those in non-collateral group (P=0.036 and 0.002 respectively). Multivariate logistic regression showed that Gensini score (OR=2.458, 95% CI=1.111 to 5.439, P=0.027) was associated with increased total maximum carotid plaque area. The optimal cut-off value for predicting the severity of CAD was 46.75 mm2 for total maximum plaque area. CONCLUSIONS: The maximum carotid plaque area can reflect the clinical severity of CAD, and it can be used as a simple noninvasive indicator of severity of coronary atherosclerosis.

Effect of Diabetes on the Assessment Role of 2-Oxoglutarate to the Severity of Chronic Heart Failure.

BACKGROUND: Serum 2-oxoglutarate can reflect the severity of chronic heart failure (CHF) in patients without diabetes. Whether this predictive role persists in type 2 diabetes mellitus (T2DM) patients is unclear. In this study, we investigated this predictive role in T2DM patients and whether 2-oxoglutarate can indicate the diastolic or systolic function of left ventricle. METHODS: One hundred eighty CHF patients (76 with T2DM) and 66 healthy controls were studied. 2-Oxoglutarate was assayed by liquid chromatography-mass spectrometry/mass spectrometry. Echocardiographic parameters, N-terminal pro-B-type natriuretic peptide (NT-proBNP) and other parameters were measured. RESULTS: 2-Oxoglutarate was increased in CHF patients with or without T2DM compared with controls (both P<0.01). Patients with a lower left ventricular ejection fraction or a higher NT-proBNP or left ventricular end-diastolic volume index had higher levels of 2-oxoglutarate (median, 18.77 µg/mL versus 11.25 µg/mL; median, 14.06 µg/ml versus 9.39 µg/ml; median, 18.06 µg/mL versus 11.60 µg/mL, all P<0.05) in nondiabetic patients but not in T2DM patients. In multiple logistic regression analysis, NT-proBNP (OR=3.445, 95% CI=1.098 to 10.816, P=0.034) and left ventricular end-diastolic diameter (OR=2.544, 95% CI=1.033 to 6.268, P=0.042) were independently associated with increased 2-oxoglutarate in nondiabetic patients. CONCLUSIONS: The levels of 2-oxoglutarate can reflect the clinical severity of CHF in nondiabetic patients but not in those with T2DM, and it can be used as a potential indicator of the systolic dysfunction of the left ventricle.

Efficacy and Safety of Renal Sympathetic Denervation on Dogs with Pressure Overload-Induced Heart Failure.

BACKGROUND: In dogs with heart failure (HF) induced by overload pressure, the role of renal sympathetic denervation (RSD) on heart failure and in the renal artery is unclear. Therefore, we investigated the efficacy and safety of RSD in dogs with pressure overload-induced heart failure. METHODS: Twenty mongrel dogs were divided into a sham-operated group, an HF group and an HF + RSD group. In the sham-operated group, the abdominal aorta was located but was not constricted, in the HF group, the abdominal aorta was constricted without RSD, and the HF+RSD group underwent RSD with constriction of the abdominal aorta after 10 weeks. Blood sampling assays, echocardiography, intravascular ultrasound (IVUS) measurement and histopathological examination were performed. RESULTS: Renal sympathetic denervation caused a significant reduction in the levels of noradrenaline (166.62±6.84 vs. 183.48±13.66 pg/ml, P<0.05), plasma renin activity (1.93±0.12 vs. 2.10±0.13 ng/mlh, P<0.05) and B-type natriuretic peptide (71.14±3.86 vs. 83.15±5.73 pg/ml, P<0.05) at eight weeks after RSD in the HF+RSD group. Compared with the HF group at eight weeks, the left ventricular internal dimension at end-diastole and end-systole were lower and the left ventricular ejection fraction was higher (all P<0.05) at eight weeks after RSD in the HF+RSD group. Intravenous ultrasound images showed no changes in the renal artery lumen, and intimal hyperplasia and vascular lumen stenosis were not observed after RSD. CONCLUSIONS: Renal sympathetic denervation could improve cardiac function in dogs with HF induced by pressure overload; RSD had no adverse influence on the renal artery.

Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna.

To yield cholinesterase (ChE) from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using forward primer 5'-CCCYGGNGCSAT GATGTG-3' and reverse primer 5'-GYAAGTTRGCCCAATATCT-3'. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a(+). The recombinant vector was transformed into Escherichia coil BL21 (DE3). Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay (IN-ELISA) was used to test the immunoreactive content of ChE (ChE-IR) in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1:22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 µg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna.

Comparison of the Target Values Established Using a Robust Statistical Method and Roche's Calibration Values.

BACKGROUND: In EQA schemes, results are usually assessed against a target value. METHODS: Outliers in each result were deleted using a robust statistical method. Mean was used as a target value if trimmed results were normally distributed; otherwise, median was used. Differences between target value and Roche's calibration value were calculated. Rates of acceptable performance evaluated by target value and Roche's calibration value were determined. RESULTS: Target values of P, GLU, urea, UA, TP, ALB, TC, TG, ALT, AST, ALP, AMY, LDH, GGT, FE, and HBDH were 1.70 mmol/L, 11.0 mmol/L, 16.8 mmol/L, 307 µmol/L, 54.0 g/L, 38.1 g/L, 4.19 mmol/L, 1.46 mmol/L, 97.7 U/L, 98.2 U/L, 206 U/L, 180 U/L, 242 U/L, 115 U/L, 35.6 µmol/L, and 255 U/L, respectively. Differences between target values and Roche's calibration values were -3.45 to 1.48%. Rates of acceptable performance evaluated by target values and Roche's calibration values were 96.5 - 100.0% and 93.0 - 100.0%, respectively. CONCLUSIONS: Target values established using a robust statistical method and Roche's calibration values were comparable.

Free triiodothyronine levels and short-term prognosis in chronic heart failure patients with type 2 diabetes.

BACKGROUND: In chronic heart failure (CHF) patients with type 2 diabetes mellitus (T2DM), the role of thyroid hormone (TH) in predicting CHF severity and prognosis is unclear. The authors therefore investigated the role of TH in predicting CHF severity and prognosis in these specific patients. METHODS: A total of 224 CHF patients (114 with T2DM) over a mean follow-up time of 6.56 ± 0.18 months were studied. TH, N-terminal pro-B-type natriuretic peptide (NT-proBNP) and other parameters were measured. RESULTS: Free triiodothyronine (FT3) levels were lower in the T2DM group compared with the nondiabetes group (P = 0.026) and higher in the New York Heart Association (NYHA) I group than in the NYHA III and IV groups (both P < 0.05). Compared with the low NT-proBNP group, the high NT-proBNP group had lower FT3 levels (P < 0.01). NT-proBNP correlated with NYHA classes (r = 0.541, P < 0.001), and inversely correlated with left ventricular ejection fraction (r = -0.431, P < 0.001) and FT3 levels (r = -0.335, P < 0.001). In multiple linear regression analysis, NT-proBNP was significantly correlated with NYHA classes (P < 0.001), left ventricular ejection fraction (P < 0.001) and FT3 (P = 0.004). Kaplan-Meier curves showed that the low FT3 group had an increased rate of short-term adverse outcomes of CHF (log rank, χ = 9.794, P = 0.002). CONCLUSIONS: FT3 levels are associated with the severity of CHF and seem to reflect short-term outcomes in CHF patients with T2DM.