Abstract Cloudy apple juice (CAJ) is popular due to its potential health benefits and pleasant flavor. However, the common used sterilization methods of CAJ by High hydrostatic pressure (HHP) or high-temperature short time (HTST) still showed many inadequacies. The present work studied the impact of HHP (400MPa/10 °C/10 min) and HTST (98 °C/50s) on CAJs made from a new cultivar Changfu 2 planted in Loess plateau area. We comparatively evaluated the microbes inactivation, enzymatic activity, phytochemical parameters, and sensory quality after the HHP and HTST treatment and during the cold storage (4 °C). Results showed that during the cold storage, both HHP and HTST treated CAJ showed a declining trend in sensory attributes like aroma, taste and overall acceptability. HTST showed a higher killing rate of bacteria, and the CAJ was detected a higher diversity of microorganisms (expressed as OTU) during the cold storage. At the end of shelf-life, Rickettsia, oxyphotobacteria, and pseudomonas were detected as the dominant genera in both HHP and HTST treated CAJs. Spearman bivariate correlate analysis determined the relationship between the bacterial profiles and enzymatic activity, phytochemical parameters, and sensory quality after the HHP and HTST treatment.

Abstract Green bactericides are excellent strategies for reducing food spoilage. Phloretin was regarded as efficient, broad-spectrum and safe preservative, which showed enormous potential in food industry. Inhibitory mechanism of phloretin on Listeria monocytogenes was systematically elucidated through integrity of cell membrane, intracellular protein content, energy level, and nucleic acid morphological character. SEM and TEM results showed minimal injury of cell membrane, which was further proved by a leakage of potassium ions, ATP and little protein and nucleic acid. There were drastic reduction of intracellular or total protein and ATP compared to cell without phloretin treatment. However, the specific activity of hexokinase and isocitrate dehydrogenase were unaffected after phloretin treatment. From these results, we concluded that phloretin’s bacteriostatic mechanism is decreasing the intracellular protein content and energy level resulting from DNA aggregation. This finding was prospective to fully illustrate the phloretin’s mechanism and develop it into a natural preservative in food industry.

Abstract The authentication of fresh apple juice (FAJ) is a significant issue. However, the main characteristics of FAJ, diluted apple juice (DAJ) with added concentrated apple juice (DAJ+CAJ), and DAJ with added sucrose (DAJ+s) are not different when using classical chemical analyses in juice authentication. In this study, isotope ratio mass spectrometry (IRMS) was used to determine the δD, δ18O, and δ13C in adulterated apple juice for juice authentication. The δD and δ18O values were decreased in adulterated juice. A linear correlation was found between δD and δ18O in apple juices with different amount of adulteration. Therefore, this linear relationship can be used to determine the amount of adulteration that took place. The δ13C value and the content of each component in real apple juice are within specific ranges. The adulteration can be recognized once the δ13C value and the content of each component in the apple juice are out of these ranges. The implementation of this method would help maintain the juice market’s stability and protect consumers’ interests.

Abstract A simple and selective liquid chromatography tandem with mass spectrometry (LC-MS/ MS) method for quantification of lobetyolin in rat plasma was developed and validated. Chromatographic separation was achieved on a Thermo ODS C18 reversed-phase column using 0.1% aqueous formic acid-methanol (50:50, v/v) in an isocratic elution mode at a flow rate of 0.4 mL.min-1. LC/MS performance was done in a positive ion ESI mode and the MS/MS transitions were monitored at m/z 419.3 [M+Na]+ → m/z 203.1 for lobetyolin and m/z 394.9 [M+Na]+ → m/z 231.9 for IS, respectively. The assay exhibited a linear dynamic range over 1.0-500 ng.mL-1 for lobetyolin in plasma. Both the precision (%RSD) and accuracy (RE%) were within acceptable criteria (<15%). Recoveries ranged from 87.0% to 95.6%, and the matrix effects were from 91.0% to 101.3%. After oral administration, the peak plasma concentration of lobetyolin was obtained as 60.1 ng.mL-1 at 1.0 h. The proposed LC-MS/MS method could be applied to a pharmacokinetic study employing 66 samples from 6 Wistar rats.

ABSTRACT Low light levels during rice (Oryza sativa L.) growing seasons may lead to significant reductions in grain yield and quality. The effect of shading stress on remobilization of nonstructural carbohydrates (NSC) from the stem plus sheath (SPS) to the grain, a process crucial to rice grain formation, was investigated. Field shading experiments were conducted in Hanyuan and Wenjiang, China, using two rice varieties (Huanghuazhan and Guichao II) subjected to shading stress after heading. Stored NSC of 60.00-159.15 g m-2 were remobilized from SPS after heading, contributing 9.63%-23.63% to grain dry weight at maturity. Shading stress resulted in the 23.99%-50.30% and 31.65%-61.33% reduction in NSC content of SPS at 20 d after heading (DAH20) and maturity stages, respectively. This contributed to the 13.80%-89.00% increase in the remobilization of stored NSC from SPS to the grain (RASN) and 15.86%-73.30% increase in remobilization percentage of removed NSC from SPS to the grain (RPRN). The effect of shading stress on remobilization differed between sites and varieties. Owing to a greater sink capacity, shading stress led to a higher RASN and RPRN from heading to DAH20 for ‘Huanghuazhan’ in Hanyuan. To compensate for photosynthate deficiency under shading stress, rice is adapted to remobilize stored NSC during early grain filling stages. Consequently, grain yield loss caused by shading could be reduced by increasing the SPS remobilization ability (especially in low light areas). Increasing NSC accumulation in SPS before heading for later remobilization may be a feasible way to alleviate rice yield loss under adverse light conditions.

Abstract A direct steam injection pretreatment was added before the microwave-assisted extraction procedure for the extraction of flavonoids from seeds of Hovenia dulcis Thunb, and the antioxidant activity of flavonoid extracts was determined. Based on the experimental results, the addition of direct steam injection simultaneously increased the extraction yield of total flavonoids compared with direct microwave-assisted extraction, and antioxidant activity of flavonoid extracts obtained by adopting the steam injection pretreatment was higher than that by direct microwave extraction alone. Myricetin was isolated from the flavonoid extracts, and the structure was identified by infrared spectroscopy (IR), electrospray ionization mass spectrometry (ESI-MS), and 1H nuclear magnetic resonance (1H-NMR). Recovery yield of total flavonoids was increased by 11.63% when the direct steam injection pretreatment was added to direct microwave extraction.

The present study aimed to investigate the in vivo inhibitory effect of histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) combined with sorafenib on human hepatocellular carcinoma HCCLM3 cells. The nude mice transplanted with HCCLM3 cells were randomly divided into control, SAHA, sorafenib and SAHA+sorafenib groups. The nude mice in the later 3 groups were intragastrically administrated with SAHA (10 mg·kg-1·day-1), sorafenib (10 mg·kg-1·day-1) and SAHA (10 mg·kg-1·day-1) combined with sorafenib (10 mg·kg-1·day-1), respectively, for successive 20 days. Finally, the inhibition rate of tumor was measured. The expressions of MEK1/2, p-ERK1/2, Cyclin D1, Bcl-2, Bax, p53, MMP-2, MMP-9 and uPA in tumor tissues were determined. Results showed that, compared with SAHA and Sorafenib groups, in SAHA+sorafenib groups the inhibition rate of tumor was significantly increased (P < 0.05), the expression levels of MEK1/2, p-ERK1/2, Cyclin D1, Bcl-2, MMP-2 and MMP-9 and uPA protein in tumor tissues were significantly decreased, respectively (P < 0.05), and the expression levels of Bax and p53 protein were significantly increased, respectively (P < 0.05). In conclusion, compared with single drug, SAHA combined with sorafenib can enhance the inhibitory effects on HCCLM3 xenografts in nude mice.

Abstract Autosomal recessive nonsyndromic hearing loss (ARNSHL) is a genetically heterogeneous neurosensory disorder, usually characterized by congenital or prelingual hearing loss. We report a Han Chinese male, born to consanguineous parents, presenting with nonsyndromic sensorineural hearing loss, whose clinical phenotype was also consistent with auditory neuropathy spectrum disorder (ANSD). After exome sequencing, a gap junction protein beta 2 gene (GJB2) c.235delC variant in the homozygous state was detected in the patient. Both parents were heterozygous for this variant, as documented by Sanger sequencing. The known pathogenic GJB2 c.235delC variant was not detected in 200 healthy controls. It is predicted to be a disease-causing alteration by generating a truncated protein p.(L79Cfs*3), disturbing the appropriate folding and/or oligomerization of connexins and leading to defective gap junction channels. This study shows that the association of homozygosity of the GJB2 c.235delC variant with ARNSHL and ANSD in a patient.

ABSTRACT MicroRNAs (miRNAs) are a class of short non-coding RNAs identified as potent regulators of gene expression. Previous studies have suggested that miRNAs are involved in mammalian spermatogenesis. Stallion fertility is an important trait for the horse breeding industry, but stallion fertility traits are largely ignored in the industry. In this study, we generated expression profiles of miRNAs in foal (immature) and stallion (mature) testes using Solexa sequencing. We identified 438 known and homologous equine miRNAs and 199 novel miRNAs which were distributed among all the chromosomes. The two developmental stages showed significant differences in miRNA expression patterns. Our result expands the horse miRNA database and provided additional information on the stallion fertility and possible spermatogenesis regulation through specific miRNAs.

BACKGROUND: Changed temperature not only threaten agricultural production, but they also affect individual biological behavior, population and community of many insects, and consequently reduce the stability of our ecosystem. Insect's ability to respond to temperature stress evolved through a complex adaptive process, thus resulting in varied temperature tolerance among different insects. Both high and low extreme temperatures are detrimental to insect development since they constitute an important abiotic stress capable of inducing abnormal biological responses. Many studies on heat or cold tolerance of ladybirds have focused on measurements of physiological and biochemical indexes such as supercooling point, higher/lower lethal temperatures, survival rate, dry body weight, water content, and developmental duration. And studies of the molecular mechanisms of ladybird responses to heat or cold stress have focused on single genes, such as those encoding heat shock proteins, but has not been analyzed by transcriptome profiling RESULTS: In this study, we report the use of Digital Gene Expression (DGE) tag profiling to gain insight into transcriptional events associated with heat- and cold-stress in C. montrouzieri. About 6 million tags (49 bp in length) were sequenced in a heat stress group, a cold stress group and a negative control group. We obtained 687 and 573 genes that showed significantly altered expression levels following heat and cold shock treatments, respectively. Analysis of the global gene expression pattern suggested that 42 enzyme-encoding genes mapped to many Gene Ontology terms are associated with insect's response to heat- and cold-stress CONCLUSIONS: These results provide a global assessment of genes and molecular mechanisms involved in heat and cold tolerance

OBJECTIVES:Our study aimed to investigate the impact of fatigue on the severity of stroke and to explore the underlying mechanisms.METHODS:Fatigued male rats underwent middle cerebral artery occlusion and the infarcted brain area was determined. Then, coagulation parameters were assessed in the fatigued group and a control group. In addition, the level of fibrinogen was determined in rats deprived of sleep for various numbers of days. To study whether interleukin-6 was involved in fibrinogen synthesis during fatigue, we also measured levels of interleukin-6 in rats deprived of sleep for various numbers of days. Furthermore, brain injury by middle cerebral artery occlusion was measured in wild-type mice, interleukin-6-/- mice and wild-type mice treated with bezafibrate.RESULTS:More severe cerebral infarction was observed in the fatigued rats, resulting in an infarct ratio of 23.4%. The infarct ratio was significantly increased in the fatigued rats compared with that in the control group (8%, p<0.05). The level of fibrinogen was increased significantly in the fatigued rats compared with that in the control group. In addition, a marked reduction in fibrinogen level was observed in the fatigued interleukin-6-/- mice compared to their wild-type counterparts, whereas no difference was observed between fatigued wild-type mice and interleukin-6-/- rats treated with recombinant human interleukin-6. The reduction in brain injury due to middle cerebral artery occlusion during fatigue was observed in interleukin-6-/- mice and wild-type mice treated with bezafibrate.CONCLUSION:Fatigue could increase stroke severity and was associated with the interleukin-6-induced expression of fibrinogen.

Pegylated interferon alpha (Peg IFN-α) in combination with ribavirin is the backbone of treatment in chronic hepatitis C (CHC). Cardiotoxicity due to interferon therapy is rare. The most frequent cardiovascular complications are arrhythmias and ischemic manifestations. Cardiomyopathy is extremely rare but can be life threatening. We present the case of a 41-year-old female patient with CHC in whom Peg IFN-α induced dilated cardiomyopathy and hypothyroidism. Chest radiography showed an enlarged and globular cardiac silhouette and pulmonary congestion. Echocardiography showed decreased left ventricular systolic function with an ejection fraction of 32% and fractional shortening of 15%. Cardiomyopathy had a complete remission after cessation of antiviral therapy with short-term heart failure medications and supportive care. Then we review the current literature about interferon induced cardiomyopathy in patients with HCV infection, as well as share our clinical experience in diagnosing and managing this rare complication.

Patients with hepatitis C virus-related decompensated cirrhosis can benefit frominterferonbased antiviral therapy, but the common complication of cytopenia is a contraindication for this treatment. Splenectomy prior to interferon therapy may alleviate this problem. To investigate whether splenectomy improves the efficacy of antiviral therapy, 13 interferonnaïve hepatitis C virus decompensated cirrhotic patients underwent splenectomy between January 2008 and January 2011, followed 1-3 months later by an interferon-based therapeutic regimen (pegylated/standard interferon-α combined with ribavirin for 48 weeks). Ten (76.9%) of the patients developed postoperative complications, which included minor portal vein thrombosis (2/13, 15.4%) and transient ascites (8/13, 61.5%). At one-month postsplenectomy, the patients showed significantly increased platelet (pre-surgery: 48.2 ± 15.9 vs. 186.0 ± 70.6×10 µL-1, p < 0.001) and leukocyte (2.1 ± 0.5 vs. 5.7 ± 1.4 × 10³ µL-1, I < 0.001) counts. Eight (61.5%) of the patients achieved sustained virological response, including all HCV genotype 2a-infected patients (4/4, 100%) and some of the genotype 1b-infected patients (4/9, 44.4%). Temporary interferon-α suspension was required for one patient to address severe intestinal infection. These results indicate that splenectomy prior to interferon-based therapy was safe and may facilitate adherence to subsequent antiviral therapy in selected HCV cirrhotic patients with portal hypertension and hypersplenism.

The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (µg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1-0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05) when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plantgrowth promoting agent.