[HbA1c comparison and diagnostic efficacy analysis of multi center different glycosylated hemoglobin detection systems].

Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R2 of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.

Waves of Change: Brain Sensitivity to Differential, not Absolute, Stimulus Intensity is Conserved Across Humans and Rats.

Living in rapidly changing environments has shaped the mammalian brain toward high sensitivity to abrupt and intense sensory events-often signaling threats or affordances requiring swift reactions. Unsurprisingly, such events elicit a widespread electrocortical response (the vertex potential, VP), likely related to the preparation of appropriate behavioral reactions. Although the VP magnitude is largely determined by stimulus intensity, the relative contribution of the differential and absolute components of intensity remains unknown. Here, we dissociated the effects of these two components. We systematically varied the size of abrupt intensity increases embedded within continuous stimulation at different absolute intensities, while recording brain activity in humans (with scalp electroencephalography) and rats (with epidural electrocorticography). We obtained three main results. 1) VP magnitude largely depends on differential, and not absolute, stimulus intensity. This result held true, 2) for both auditory and somatosensory stimuli, indicating that sensitivity to differential intensity is supramodal, and 3) in both humans and rats, suggesting that sensitivity to abrupt intensity differentials is phylogenetically well-conserved. Altogether, the current results show that these large electrocortical responses are most sensitive to the detection of sensory changes that more likely signal the sudden appearance of novel objects or events in the environment.

Methionine and arginine supplementation alter inflammatory and oxidative stress responses during lipopolysaccharide challenge in bovine mammary epithelial cells in vitro.

Mastitis, inflammation of the udder, is one of the most common diseases hampering milk yield of dairy cows. Methionine (Met) and arginine (Arg) are key nutrients with potential to regulate inflammation and oxidative stress. The aim of this study was to evaluate the effect of increased supply of Met and Arg on mRNA and protein abundance associated with innate immune response and redox balance during lipopolysaccharide (LPS) stimulation in primary bovine mammary epithelial cells (BMEC). Primary BMEC (n = 4 replicates per treatment) were pre-incubated for 12 h in media with the following amino acid combinations: ideal profile of amino acids (control; Con), increased Met supply (incMet), increased Arg supply (incArg), and increased supply of Met and Arg (incMetArg). Subsequently, cells were challenged with or without LPS (1 µg/mL) and incubated for 6 h. Data were analyzed as a 2 × 2 × 2 factorial using the MIXED procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). The downregulation of SLC36A1 and SLC7A1 mRNA abundance induced by LPS was attenuated in the incArg cultures. Although challenge with LPS led to lower abundance of proteins related to the antioxidant response (NFE2L2, NQO1, GPX1), lower levels of ATG7, and lower mRNA abundance of GPX3, we found little effect in cultures with incMet or incArg. Cultures with incMet, incArg, or incMetArg led to attenuation of the upregulation of SOD2 and NOS2 induced by LPS. Abundance of phosphorylated p65 (RELA) was greater after LPS stimulation, but the response was attenuated in cultures with incMet. The greater ratio of pRELA to total RELA in responses to LPS was also attenuated in cultures with incMetArg. The greater mRNA abundance of the proinflammatory cytokine IL1B induced by LPS was attenuated in cultures with incMet, and the same trend induced by LPS on CXCL2 was also alleviated in cultures with incArg. Overall, the data suggest that greater supply of Met and Arg alleviated the proinflammatory responses triggered by LPS through controlling the abundance of proinflammatory cytokines and chemokines and activity of NF-κB. Little benefit on oxidative stress induced by LPS challenge in BMEC was detected with greater supply of Met and Arg.

Neurobiological mechanisms of TENS-induced analgesia.

Pain inhibition by additional somatosensory input is the rationale for the widespread use of Transcutaneous Electrical Nerve Stimulation (TENS) to relieve pain. Two main types of TENS produce analgesia in animal models: high-frequency (∼50-100 Hz) and low-intensity 'conventional' TENS, and low-frequency (∼2-4 Hz) and high-intensity 'acupuncture-like' TENS. However, TENS efficacy in human participants is debated, raising the question of whether the analgesic mechanisms identified in animal models are valid in humans. Here, we used a sham-controlled experimental design to clarify the efficacy and the neurobiological effects of 'conventional' and 'acupuncture-like' TENS in 80 human volunteers. To test the analgesic effect of TENS we recorded the perceptual and brain responses elicited by radiant heat laser pulses that activate selectively Aδ and C cutaneous nociceptors. To test whether TENS has a long-lasting effect on brain state we recorded spontaneous electrocortical oscillations. The analgesic effect of 'conventional' TENS was maximal when nociceptive stimuli were delivered homotopically, to the same hand that received the TENS. In contrast, 'acupuncture-like' TENS produced a spatially-diffuse analgesic effect, coupled with long-lasting changes both in the state of the primary sensorimotor cortex (S1/M1) and in the functional connectivity between S1/M1 and the medial prefrontal cortex, a core region in the descending pain inhibitory system. These results demonstrate that 'conventional' and 'acupuncture-like' TENS have different analgesic effects, which are mediated by different neurobiological mechanisms.

Metabonomic analysis of toxic action of long-term low-level exposure to acrylamide in rat serum.

This study assessed the effects of long-term, low-dose acrylamide (AA) administration in rats using ultra-performance liquid chromatography-mass spectrometry. Forty male Wistar rats were randomly divided into the following four groups: control, low-dose AA (0.2 mg/kg BW), middle-dose AA (1 mg/kg BW), and high-dose AA (5 mg/kg BW). AA was administered to rats via drinking water ad libitum. After 16-week treatment, rat serum was collected for metabonomic analysis. Biochemical tests were further conducted to verify metabolic alterations. Eleven metabolites were identified with significant changes in intensities (increased or reduced) as a result of treatment. These metabolites included citric acid, pantothenic acid, isobutyryl-l-carnitine, eicosapentaenoic acid, docosahexaenoic acid, sphingosine 1-phosphate, LysoPC(20:4), LysoPC(22:6), LysoPE(20:3), undecanedioic acid, and dodecanedioic acid. Results indicate that chronic exposure to AA at no observed adverse effect level does not exert a toxic effect on rats at the body metabolism level. AA disturbed the metabolism of lipids and energy, affected the nervous system of rats, and induced oxidative stress and liver dysfunction.

[Study on the overall implementation status of the National Demonstration Areas for Comprehensive Prevention and Control of Non-communicable Diseases].

Objective: To understand the current overall status of implementation on the National Demonstration Areas of Comprehensive Prevention and Control of Non-communicable Diseases. Methods: According to the scheme design of the questionnaires, all the National Demonstration Areas were involved in this study. For each National Demonstration Areas, eight departments were selected to complete a total of 12 questionnaires. Results: Scores related to the implementation of the National Demonstration Areas accounted for 71.8% of the total 170 points. Based on the scores gathered from this study, the 23-items-index-system that represented the status of project implementation was classified into seven categories. Categories with higher percentile scores would include: monitoring (88.0%), safeguard measures (75.0%), health education and health promotion (75.0%). Categories with lower percentile scores would include: the national health lifestyle actions (67.7%), community diagnosis (66.7%), discovery and intervention of high-risk groups (64.7%), and patient management (60.9%). There were significant differences noticed among the eastern, central and western areas on items as safeguard measures, health education/promotion, discovery and intervention of high-risk groups. In all, the implementation programs in the eastern Demonstration Areas seemed better than in the central or western regions. As for the 23 items, five of the highest scores appeared on policy support, mortality surveillance, tumor registration, reporting system on cardiovascular/cerebrovascular events, and on tobacco control, respectively. However, the lowest five scores fell on healthy diet, patient self-management program, oral hygiene, setting up the demonstration units and promotion on basic public health services, respectively. The overall scores in the eastern region was higher than that in the central or the western regions. The scores in the central and western regions showed basically the same. Conclusions: The overall status of implementation on the National Demonstration Areas was satisfactory. Future attention should be focusing on patient management as well as discovery and intervention of high-risk groups, which also presented the lowest scores, in this survey.

[The protective effect of Xuebijing on paraquat-induced HK-2 cells apoptosis and the underlying mechanisms].

Objective: To investigate the protective effect and mechanism of Xuebijing (XBJ) on paraquat (PQ) -induced apoptosis in Human kidney cell line-2 (HK-2) cells. Methods: Routinely cultured HK-2 cells, (1) Cell growth inhibition experiment after PQ and XBJ intervention: PQ was divided into 0、200、400、800、1600 and 3200 µmol/L PQ groups, and the cell survival rate was detected after intervening 24、48 and 72 h. XBJ was divided into 0、5、10、20、40 mg/ml XBJ groups, and the cell survival rate was detected after intervening 24、48 and 72 h.To determine the rational drug concentration and the duration of action of XBJ and PQ. (2) PQ-induced HK-2 cell growth inhibition experiment antagonized by XBJ: The cells were divided into normal control group, PQ group (800 µmol/L) and PQ+XBJ group (The cells were pretreated with 5、10 and 20 mg/ml XBJ for 1 h, then cultured with PQ of 800 µmol/L) , After cultured 24 h、48 h and 72 h separately, the cell survival rate was detected. (3) HK-2 cells were divided into normal control group、PQ group (800 µmol/L PQ cultured for 24 h) 、PQ+XBJ group (pretreated with 10 mg/ml XBJ for 1 h, and then 800 µmol/L PQ cultured for 24 h) and XBJ group (10 mg/ml XBJ cultured 24 h). The apoptosis of cells was detected by flow cytometry. The protein expression of Bcl-2 and BAX in each group was detected by Western blotting. The expressions of caspase-3 and caspase-9 were detected by caspase-3 and caspase-9 activity kit active. Results: (1) PQ could significantly reduced the survival rate of HK-2 cells and showed time and concentration dependence. The survival rate of HK-2 cells was about 55% after 800 µmol/L PQ contacted 24 h, XBJ under 20 mg/ml was no significant effect on the survival rate of HK-2 cells after cultured 72 h. (2) Compared with the PQ group, the survival rate of HK-2 cells of PQ+XBJ group was significantly increased (P<0.05). (3) Compared with the normal control group, the cell apoptosis rate of PQ group was significantly increased (P<0.05). Compared with the PQ group, the cell apoptosis rate of PQ+XBJ group was significantly decreased (P<0.05). (4) Compared with the normal control group, Bcl-2 protein expression in PQ group was significantly decreased and BAX protein expression in PQ group was significantly increased (P<0.05) ; compared with PQ group, Bcl-2 protein expression in PQ+XBJ group was significantly increased, BAX protein expression in PQ+XBJ group was significantly decreased (P<0.05). (5) Compared with the normal control group, the activities of caspase-3 and caspase-9 in PQ group were significantly increased (P<0.05). Compared with PQ group, the activities of caspase-3 and caspase-9 in PQ+XBJ group were decreased significantly (P<0.05) . Conclusion: XBJ (10 mg/ml) has obvious protective effect on HK-2 cell injuried by PQ (800 µmol/L) , It can improve the survival rate of cells through reducing the apoptosis of HK-2 cells which induced by PQ.

Oral immunisation with Taishan Pinus massoniana pollen polysaccharide adjuvant with recombinant Lactococcus lactis-expressing Proteus mirabilis ompA confers optimal protection in mice.

BACKGROUND: Proteus mirabilis poses a critical burden on the breeding industry, but no efficient vaccine is available for animals. METHOD: A recombinant Lactococcus lactis expressing the ompA of P. mirabilis was used to develop a vaccine. The mucosal and systemic immune responses of the recombinant vaccine were evaluated in mice after oral immunisation. The inhibition on P. mirabilis colonisation of vaccines was also determined. Moreover, Taishan Pinus massoniana pollen polysaccharides (TPPPS) were used as adjuvants to examine the immunomodulatory effects. RESULTS: The pure recombinant L. lactis vaccine significantly induced the production of specific IgA and IgG, IL-2, IL-4, IFN-γ, and T lymphocyte proliferation, and the immunised mice exhibited significant resistance to P. mirabilis colonisation. Notably, the TPPPS adjuvant vaccines induced higher levels of immune responses than the pure L. lactis. CONCLUSIONS: The L. lactis as a vaccine vehicle combined with TPPPS adjuvant provides a feasible method for preventing P. mirabilis infection.

The addition of protein-bound amino acids in low-protein diets improves the metabolic and immunological characteristics in fifteen- to thirty-five-kg pigs.

This study was to evaluate the effects of supplementation of AA form (crystalline vs. protein bound) in low-protein diets on growth, metabolic, and immunological characteristics of pigs. A total of 80 barrows (PIC 327 × 1050; 15.57 ± 0.13 kg BW and 48 ± 2 d of age), housed in 4 pigs per pen with 5 pens per treatment, were assigned to 4 dietary treatments of 17, 15, and 13% CP and 13% CP plus casein for 28 d. The crystalline AA were supplemented to meet the requirement of indispensable AA in pigs. Results showed that pigs fed the 13% CP diet or the 13% CP plus casein diet had lower ( < 0.01) ADG and ADFI and a greater ( < 0.01) feed:gain ratio than pigs fed the 17% CP or 15% CP diets over the 4-wk study period. Compared with other diets, pigs fed the 13% CP diet had decreased concentrations of plasma urea nitrogen, albumin ( < 0.01), and mRNA expressions of Toll-like receptor 4 (), nuclear factor kappa B (; < 0.05), and Toll-interacting protein (; < 0.01) in the ileum and also increased activity of plasma glutamate-pyruvate transaminase ( < 0.05) and concentrations of IL-1ß ( < 0.05) and tumor necrosis factor-α ( < 0.01); however, these characteristics were partly normalized by feeding the 13% CP plus casein diet. Furthermore, the plasma concentration of insulin-like growth factor 1 (IGF-1; < 0.01) and mRNA expressions of protein kinase B (), mammalian target of rapamycin (), and ribosomal protein S6 kinase () in longissimus muscle were increased ( < 0.05) in pigs fed the 13% CP plus casein diet relative to pigs fed the 17% CP or 15% CP diets. In summary, reducing dietary CP level from 17% to 15% had no effect on growth, metabolic, and immunological characteristics of 15- to 35-kg pigs. A further reduction of dietary CP level up to 13% would lead to poor growth performance, but metabolic and immunological characteristics were partly normalized using protein-bound AA to replace synthesized AA in the 13% CP diet.

Sensitive and rapid detection of Pectobacterium atrosepticum by targeting the gyrB gene using a real-time loop-mediated isothermal amplification assay.

UNLABELLED: This study reports the development of a real-time, loop-mediated isothermal amplification (RealAmp) assay for the detection of Pectobacterium atrosepticum (P. atrosepticum). A phylogenetic tree was constructed based on the gyrB gene of P. atrosepticum and related species. Pectobacterium atrosepticum from different sources can be clustered in the same branch with 100% support rate. The RealAmp primers targeting the gyrB gene of P. atrosepticum worked most efficiently at 61·0°C. Compared with 55 related bacterial strains, the eight P. atrosepticum strains displayed positive reaction in the RealAmp assay. The melting temperature (Tm) of P. atrosepticum amplified products was about 85·0°C. The detection limit of the RealAmp assay for the detection of P. atrosepticum in pure culture was approx. 3 CFU reaction(-1) . The detection limit of the RealAmp assay for the detection of P. atrosepticum in artificially contaminated samples was 22 CFU reaction(-1) . The detection rate of the RealAmp assay for the detection of potato tubers was 28·5-32·0% higher than that of the conventional PCR. In summary, a specific, sensitive and rapid RealAmp assay based on the gyrB gene of P. atrosepticum, which can be easily performed and real-time monitored, was established. SIGNIFICANCE AND IMPACT OF THE STUDY: Potato blackleg caused by Pectobacterium atrosepticum (P. atrosepticum) which is mainly transmitted through the seed potato leads to the decline in potato production. To reduce yield loss, rapid detection of P. atrosepticum in seed potato remains essential. Based on the gyrB gene of P. atrosepticum, species-specific primers were designed. A real-time, loop-mediated isothermal amplification (RealAmp) assay was established for the detection of P. atrosepticum. The RealAmp assay is a specific, rapid and sensitive method for P. atrosepticum detection. Therefore, it provides an effective diagnosis of potato blackleg in both the growing and stored potato.

Effects of low-protein diets supplemented with indispensable amino acids on growth performance, intestinal morphology and immunological parameters in 13 to 35 kg pigs.

The objective of this study was to determine if a moderate or high reduction of dietary CP, supplemented with indispensable amino acids (IAA), would affect growth, intestinal morphology and immunological parameters of pigs. A total of 40 barrows (initial BW=13.50±0.50 kg, 45±2 day of age) were used in a completely randomized block design, and allocated to four dietary treatments containing CP levels at 20.00%, 17.16%, 15.30% and 13.90%, respectively. Industrial AA were added to meet the IAA requirements of pigs. After 4-week feeding, blood and tissue samples were obtained from pigs. The results showed that reducing dietary CP level decreased average daily gain, plasma urea nitrogen concentration and relative organ weights of liver and pancreas (P<0.01), and increased feed conversion ratio (P<0.01). Pigs fed the 13.90% CP diet had significantly lower growth performance than that of pigs fed higher CP at 20.00%, 17.16% or 15.30%. Moreover, reducing dietary CP level decreased villous height in duodenum (P<0.01) and crypt depth in duodenum, jejunum and ileum (P<0.01). The reduction in the dietary CP level increased plasma concentrations of methionine, alanine (P<0.01) and lysine (P<0.05), and decreased arginine (P<0.05). Intriguingly, reducing dietary CP level from 20.00% to 13.90% resulted in a significant decrease in plasma concentration of IgG (P<0.05), percentage of CD3+T cells of the peripheral blood (P<0.01), also down-regulated the mRNA abundance of innate immunity-related genes on toll-like receptor 4, myeloid differentiation factor 88 (P<0.01) and nuclear factor kappa B (P<0.05) in the ileum. These results indicate that reducing dietary CP level from 20.00% to 15.30%, supplemented with IAA, had no significant effect on growth performance and had a limited effect on immunological parameters. However, a further reduction of dietary CP level up to 13.90% would lead to poor growth performance and organ development, associated with the modifications of intestinal morphology and immune function.

The effects of enzyme supplementation on performance and digestive parameters of broilers fed corn-soybean diets.

The study was performed to evaluate the effects of enzyme supplementation on performance and digestive parameters of broilers fed corn-soybean diets from 1 to 21 d of age. A total of 480 one-day-old Cobb broilers were allocated to 1 of 4 treatments, with 6 replicate pens per treatment and 20 birds per pen. The experiment consisted of a 2 × 2 factorial arrangement of treatments with 2 dietary ME levels (high ME, energy 1, 12.13 MJ/kg or low ME, energy 2, 11.92 MJ/kg) and 2 levels of supplemental enzyme (including xylanase, 1,800 IU/g, ß-glucanase, 500 IU/g, and α-amylase, 800 U/g; 0 or 0.1% of diet). Enzyme supplementation had no effect on average daily weight gain, feed intake, and feed:gain. However, enzyme supplementation decreased the relative weight of the pancreas (d 7 and 21) in broilers fed the high ME diet. Low dietary ME level increased pancreatic lipase (d 7, P = 0.015), trypsin (d 14, P = 0.01; d 21, P = 0.014), amylase (d 21, P = 0.027), and pepsin (d 7, P = 0.001; d 21, P = 0.042) activity, but reduced pancreatic lipase (d 14, P = 0.03; d 21, P = 0.004) and amylase (d 14, P = 0.027) activity. Enzyme supplementation resulted in an increase in pancreatic amylase (d 7, P = 0.023), trypsin (d 7, P = 0.02; d 21, P = 0.004), lipase (d 21, P = 0.001), pepsin (d 7, P = 0.001; d 14, P = 0.004; d 21, P = 0.001), and maltase (d 14, P = 0.011, in ileum) activity. Moreover, broilers fed low dietary ME and enzyme supplementation diets had an increase in pancreatic lipase (d 21, P = 0.001) and pepsin (d 7, P = 0.001) activity. Low ME diets reduced jejunum villus height and jejunum and ileum crypt depth (d 7, 21). However, enzyme supplementation, especially enzyme supplementation in low ME diets, increased jejunum and ileum villus height and villus surface area. This suggested enzyme supplemented with low ME diet might be more effective to improve the activity of digestive enzymes and the absorptive capacity of the small intestine.

Effects of dietary supplementation of quercetin on performance, egg quality, cecal microflora populations, and antioxidant status in laying hens.

Plant polyphenols, especially flavonoids, are of great interest due to their wide range of biological activities. Quercetin, a ubiquitous flavonoid, is known to have antioxidant and antibacterial effects. In this study, we investigated the effect of quercetin on performance, egg quality, cecal microflora populations, and antioxidant status in laying hens. Two hundred forty 28-wk-old Hessian laying hens, with an average laying rate of lay 85% at the start, were randomly allotted to 4 treatments and fed 1 of 4 diets (negative control, 0.2, 0.4, and 0.6 g of quercetin/kg of diet) for 8 wk. Layer performance responses, egg quality parameters, cecal microflora populations and antioxidant status were measured at the end of the experiment. Results showed that feed conversion decreased as the quercetin level increased. Laying rate had a quadratic correlation with the level of quercetin (P = 0.056) and was maximized by the supplementation level of 0.2 g/kg of diet. However, no significant quercetin effect was observed on egg quality. Regression analysis showed that the population of total aerobes and coliforms decreased and the population of Bifidobacteria increased as the level of quercetin increased. Regression analysis also showed the activities of Cu-Zn-superoxide dismutase increased as the level of quercetin increased (P < 0.05). Results of the study suggest that the appropriate level of supplementation is 0.367 to 0.369 g of quercetin/kg of feed based on the improvement of laying rate (with 88.55 as maximum value) and feed conversion (with 2.0725 as minimum value). Our observations provided further evidence that dietary supplementation of quercetin improved performance by modulation of intestinal environment and liver superoxide dismutase content in laying hens. Quercetin has the potential as functional feed additive in animal production.

Effect of quercetin on performance and egg quality during the late laying period of hens.

1. At the end of the peak laying period for fowl, both performance and egg quality decrease markedly. Counteracting either or both of these could, therefore, have significant positive economic implications. Quercetin, a typical representative of the flavonol compounds, has a variety of biological functions. However, there are no reported findings on its use as a feed additive. 2. In this study, we investigated the effects of quercetin on laying rate, egg quality and blood traits associated with egg quality in laying hens (Hessain) during the late laying period and explored the possibility of using quercetin as a functional feed additive. 3. The laying hens (n = 240; 39 weeks old) were randomly assigned to 4 treatments consisting of 6 replicates (n = 10) and were fed with diets containing quercetin at 0, 0.2, 0.4 or 0.6 g/kg. 4. Results showed that laying rate was increased and feed-egg ratio was decreased significantly by 0.2 and 0.4 g/kg quercetin. Compared with controls, Haugh unit, eggshell strength, eggshell thickness and yolk protein were increased, but yolk cholesterol was decreased by quercetin. 5. In conclusion, quercetin was beneficial in improving performance and egg quality. The recommended concentration of quercetin is 0.4 g/kg of the basal diet. At this concentration, increased laying rate, improved egg quality and reduced yolk cholesterol can be expected.

Inhibitory effects of tea extract on aflatoxin production by Aspergillus flavus.

UNLABELLED: Aflatoxins, one of the most carcinogenic substances, have been implicated as a potential threat to the safety of tea beverages. In this study, we studied the inhibitory effects of the aqueous extracts from several Chinese traditional teas, such as green tea, black tea, flower tea, raw Puer tea (naturally fermented Puer tea) and Puer tea (inoculated Puer tea), on the growth and aflatoxin production of Aspergillus flavus. All the tested extracts inhibited the production of aflatoxin B1, whereas they did not inhibit mycelial growth of A. flavus. Considering the highest inhibitory effect of Puer tea extract on aflatoxin production, a semi-quantitative RT-PCR was designed to detect its impacts on the expression of genes responsible for the regulation of aflatoxin synthesis. The results showed that the transcriptions of both aflS and aflR were down-regulated to undetectable levels by the addition of Puer tea extract. This study indicated that most tea contained molecules inhibitory to aflatoxin production, which were very important factors for the risk assessment of tea exposed to aflatoxin. Some tea extracts could be developed as antiaflatoxin agents in food preservation. SIGNIFICANCE AND IMPACT OF THE STUDY: Recently, safety concerns of the popular Puer tea have arisen because of aflatoxin contamination. In this study, we analysed the inhibitory effect of 30 tea aqueous extracts on the growth and aflatoxin production of Aspergillus flavus. Our results indicated that most tea inhibited aflatoxin production by down-regulating the transcription of aflR and aflS. The findings could contribute to the safety assessment of tea exposed to aflatoxin and provide some useful data concerning a new approach for controlling aflatoxin contamination.

Intense nanosecond pulsed electric fields promote cancer cell apoptosis through centrosome-dependent pathway involving reduced level of PLK1.

BACKGROUND: Intense nanosecond pulsed electric fields (nsPEFs) have been known to promote apoptosis without physically changing membrane structure or damaging morphology of tumor cells. To determine the contribution of centrosome to the progression of apoptosis by nsPEFs, HeLa cells were exposed to high intensity (6 kV/cm) nsPEFs (8-32 ns) in normal culture condition and cell biology and molecular parameters of cells were investigated. MATERIALS AND METHODS: Apoptotic cell death was identified by TUNEL assay after being exposed to the nsPEFs with various pulse durations, while immunofluorescent staining was performed to detect the number and distribution of centrosomes. To clarify whether nsPEFs-induced centrosome over-duplication is the consequence of DNA damage, we used comet assay to detect simultaneous DNA damage. And additionally Western Blot was used to detect PLK1 protein level to explore the correlation between apoptotic cell death and nsPEFs-induced centrosome over-duplication. Correlation between nsPEFs and molecular parameters was statistically analyzed. RESULTS: NsPEFs induced a clear apoptosis reaching a maximum at 24ns, 24h after exposure (p < 0.05), where DNA fragmentation and over-duplicated centrosomes were observed. This apoptosis may be promoted in a time- and pulse duration-dependent manner. Polo-like kinase (PLK1) protein levels were significantly decreased by such nsPEFs (p < 0.05). Control treatment without the nsPEFs did not cause any damage to the cultured HeLa cells. CONCLUSIONS: Intense nsPEFs promote cell apoptosis through a centrosome-mediated pathway involving a reduction in the level of PLK1, which may provide new therapeutic targets for human cancer treatment.

Neuroprotection of Scutellarin is mediated by inhibition of microglial inflammatory activation.

Inhibition of microglial over-reaction and the inflammatory processes may represent a therapeutic target to alleviate the progression of neurological diseases, such as neurodegenerative diseases and stroke. Scutellarin is the major active component of Erigeron breviscapus (Vant.) Hand-Mazz, a herbal medicine in treatment of cerebrovascular diseases for a long time in the Orient. In this study, we explored the mechanisms of neuroprotection by Scutellarin, particularly its anti-inflammatory effects in microglia. We observed that Scutellarin inhibited lipopolysaccharide (LPS)-induced production of proinflammatory mediators such as nitric oxide (NO), tumor necrosis factor α (TNFα), interleukin-1ß (IL-1ß) and reactive oxygen species (ROS), suppressed LPS-stimulated inducible nitric oxide synthase (iNOS), TNFα, and IL-1ß mRNA expression in rat primary microglia or BV-2 mouse microglial cell line. Scutellarin inhibited LPS-induced nuclear translocation and DNA binding activity of nuclear factor κB (NF-κB). It repressed the LPS-induced c-Jun N-terminal kinase (JNK) and p38 phosphorylation without affecting the activity of extracellular signal regulated kinase (ERK) mitogen-activated protein kinase. Moreover, Scutellarin also inhibited interferon-γ (IFN-γ)-induced NO production, iNOS mRNA expression and transcription factor signal transducer and activator of transcription 1α (STAT1α) activation. Concomitantly, conditioned media from Scutellarin pretreated BV-2 cells significantly reduced neurotoxicity compared with conditioned media from LPS treated alone. Together, the present study reported the anti-inflammatory activity of Scutellarin in microglial cells along with their underlying molecular mechanisms, and suggested Scutellarin might have therapeutic potential for various microglia mediated neuroinflammation.

Antagonistic effects of two herbs in Zuojin Wan, a traditional Chinese medicine formula, on catecholamine secretion in bovine adrenal medullary cells.

In order to research the target of superior efficacy and lesser side effects, combination of herbal materials has been applied to phytotherapy for thousands of years in China and some other countries. Zuojin Wan (ZJW), a famous traditional Chinese medicine formula, is used in treating gastric diseases in China. It is composed of two herbs, Rhizoma Coptidis (RC) and Fructus Evodiae (FE) in the ratio of 6: 1(w/w). In the present study, we examined the effects of ZJW, RC, FE and active components isolated from these herbs on catecholamine (CA) secretion and intracellular calcium ([Ca(2+)](i)) in cultured bovine adrenal medullary cells. Extracts of ZJW and RC and berberine, palmatine and jatrorrhizine, components of RC, all inhibited CA secretion and rise in [Ca(2+)](i) induced by acetylcholine (ACh), veratridine (Ver) and/or 56 mM K(+). On the other hand, extract of FE, evodiamine and rutaecarpine, components of FE, stimulated CA secretion and rise in [Ca(2+)](i) induced by ACh. Furthermore, different proportions of RC and FE caused different responses in CA secretion. The present findings suggest that two herbs in ZJW have opposite effects, i.e., inhibitory effect of RC and stimulatory effect of FE, on CA secretion induced by acetylcholine in cultured bovine adrenal medullary cells.

Four weeks high fat feeding induces insulin resistance without affecting dopamine release or gene expression patterns in the hypothalamus of C57Bl6 mice.

Obesity is associated with diminished dopaminergic neurotransmission. It remains unclear whether this is a cause or a consequence of the obese state. We hypothesized that high fat feeding, a well known trigger of obesity in diet sensitive mice, would blunt dopaminergic neurotransmission prior to the development of insulin resistance. We monitored in vivo dopamine release in the dorsomedial region of the hypothalamus, and determined hypothalamic gene expression patterns of dopamine receptors 1 and 2 (DRD1 and 2), tyrosine hydroxylase (TH) and the dopamine transporter (DAT) in C57Bl6 mice maintained on a high fat diet for 4 weeks. Also, a hyperinsulinemic euglycemic clamp was performed to evaluate the metabolic status of the mice. Mice maintained on a low fat diet served as controls. The high fat diet did not alter dopamine release in the dorsomedial hypothalamus of fed or fasted mice or the dopaminergic response to refeeding. Furthermore, gene expression levels of DRD1, DRD2, TH and DAT were not affected by high fat feeding. However, the high fat diet did hamper insulin action as evidenced by diminished glucose disposal during hyperinsulinemia (p<0.05). We show here that short term high fat feeding does not affect dopaminergic neurotransmission in the hypothalamus, whereas it does impair insulin action. This suggests that reduced dopaminergic neurotransmission in the hypothalamus of obese animal models is due to mechanism(s) that are not directly triggered by diet composition.

Schoepfin A, B, C: three new chalcone C-glycosides from Schoepfia chinensis.

Three new chalcone C-glycosides named schoepfin A, B, C (1-3), together with three known compounds 4,2',4'-trihydroxy-3'-C-beta-D-glucosylchalcone (4), nothofagin (5) and hemiphloin (6) were isolated from ethanolic extract of the bark of Schoepfia chinensis Gardn. et Champ (Olacaceae). Their structures were determined mainly by spectroscopic techniques including 2D-NMR (HMBC, HMQC) and MS experiments.