Effect of atmospheric cold plasma treatment on antioxidant activities and reactive oxygen species production in postharvest blueberries during storage.

BACKGROUND: Blueberry is universally acknowledged as a kind of berry rich in antioxidants. Cold plasma, an emerging non-thermal treatment technology, has been proved to be able to maintain or improve the antioxidant level while inactivating the microorganisms on the surface of fruits and vegetables. Postharvest blueberries were treated with atmospheric cold plasma (ACP; 12 kV, 5 kHz) for 0 s (Control), 30 s (ACP-30), 60 s (ACP-60), and 90 s (ACP-90) in this study, and the effects of ACP on the antimicrobial properties, antioxidant activities, and reactive oxygen species (ROS) production were investigated during storage at 4 ± 1 °C for 40 days. RESULTS: Total aerobic bacteria and mold populations on ACP-treated blueberries decreased significantly in a time-dependent manner (P < 0.05), and decreased by 0.34-1.24 and 0.57-0.87 log10 CFU g-1 respectively on ACP-60-treated blueberries during storage. The decay rate of blueberries was decreased by 5.8-11.7% and the decrease of blueberry firmness was slowed down by ACP-60. But the total phenol, anthocyanin, and ascorbic acid contents increased, and superoxide dismutase, catalase, and peroxidase activities were enhanced in ACP-treated blueberries. The free radical scavenging activity and total antioxidant capacity (T-AOC) were enhanced. Hydrogen peroxide (H2 O2 ) and superoxide anion (O2 - ) production rates declined by 27.3% and 41.3% at day 40 of storage, respectively. CONCLUSION: It is suggested that ACP may be a promising non-thermal treatment technology for postharvest sterilization and preservation of blueberry under suitable conditions. © 2020 Society of Chemical Industry.

Effect of Th17 and Treg axis disorder on outcomes of pulmonary arterial hypertension in connective tissue diseases.

This prospective cohort study is to verify the hypothesis that the balance of Th17 and Treg cells frequencies in the peripheral circulation is disturbed in patients with varying degrees of connective tissue diseases-associated pulmonary arterial hypertension (CTD-aPAH) and to prove the influence of Th17/Treg imbalance on prognosis. We detected the frequencies and absolute counts of Th17 and Treg cells and related serum cytokines secretion and expressions of key transcription factors in 117 patients with connective tissue diseases (CTD), 53 patients with CTD-aPAH, and 48 healthy volunteers. Moreover, the median value according to levels of Th17/Treg ratios in patients with CTD-aPAH was chosen as basis of group division for survival analysis. CTD-aPAH patients revealed significant increase in peripheral Th17 cells, Th17-related cytokines, and ROR γt mRNA levels. They also presented a significant decrease in Treg cells, Treg-related cytokines, and Foxp3 mRNA levels as compared with CTD patients and healthy controls. More importantly, the Th17/Treg ratio was significantly related to the severity and prognosis of CTD-aPAH. This study indicated that the Th17/Treg axis disorder plays a critical role in CTD-aPAH. Furthermore, the dynamic balance between Th17 and Treg cells was likely to influence prognosis of patients with CTD-aPAH.

Analysis and identification of key anti-inflammatory molecules in Eerdun Wurile and exploration of their mechanism of action in microglia.

Ethnomedicine Eerdun Wurile (EW) can significantly promote poststroke neuro-recovery through modulation of microglia polarization. Fraction 4-6 (F4-6) isolated from EW via serial fractionation inhibits the expression of pro-inflammatory genes in LPS stimulated microglia. However, the key active molecules of F4-6 have not been identified. Herein, we identified alantolactone (Ala) and dehydrodiisoeugenol (Deh) as the active anti-inflammatory components of F4-6 by UPLC-qTof MS analysis. We confirmed that, F4-6, Ala, Deh and mixture of Ala and Deh (Mix) downregulate the expression of several pro-inflammatory genes including Ccl2, Cox2 and Il6 in LPS-treated microglia in a similar pattern. At the same time upregulate the expression of anti-inflammatory genes including Hmox1, Tgfß, Igf1 and Creb1. Moreover, the conditioned culture media obtained from F4-6 treated microglia significantly enhanced proliferation of N2a cells, and promoted neurite outgrowth possibly through upregulation of Nefh and Dlg4. Mechanistically, F4-6 strongly downregulated the expression of NF-κB p65, while also inhibiting the nuclear translocation of p65, leading to the suppression of transcription of pro-inflammatory genes initiated by NF-κB. Collectively, our data identified and quantified the key chemicals of EW and provide insights into the optimization of the herbal composition for neuroprotection.

Ventilator-associated pneumonia in patients with cerebral hemorrhage: Impact on mortality and microbiological characterization.

Ventilator-associated pneumonia (VAP) is a major complication among critically ill patients who depend on mechanical ventilation. Few reports have focused on intracerebral hemorrhage patients with VAP. Our main objective was to investigate the bacteria distribution characteristics and the impact of ventilator-associated pneumonia mortality in critical cerebral hemorrhage patients. This retrospective study included 89 cases of cerebral hemorrhage patients with VAP admitted to the ICU of Huashan Hospital. We used the chi-square test to compare qualitative variables and Student's t-test to compare means between groups of normally distributed quantitative variables. Multiple logistic regression analysis was used to assess mortality-independent predictors in the ICU. A total of 42% patients with cerebral hemorrhage were diagnosed with VAP in the ICU during the study period, and the mortality rate was 18%. Acinetobacter baumannii (n=58), Klebsiella pneumoniae (n=52), and Pseudomonas aeruginosa (n=21) were the most common pathogenic bacteria. Blood volume >30ml, tracheal ventilation mode and head of bed elevation were independent factors associated with increased mortality. Glasgow Coma Scale (GCS), Acute Physiology and Chronic Health Evaluation II (APACHE II) score and the time from bleeding to intubation were other potentially important factors. While the number of infecting bacteria may not be directly related to death, it can increase antibiotic consumption and length of intensive care unit (ICU) stays. Blood volume >30ml, tracheal ventilation mode and head of bed elevation were directly related to the death of critical cerebral hemorrhage patients with ventilator-associated pneumonia.

Characterization and expression of gamma-interferon-inducible lysosomal thiol reductase (GILT) gene in amphioxus Branchiostoma belcheri with implications for GILT in innate immune response.

An amphioxus cDNA, AmphiGILT, encoding GILT protein was isolated from the gut cDNA library of Branchiostoma belcheri. It codes for a deduced protein of 254 amino acids, which has all the main features typical of GILT proteins including the signature sequence CQHGX(2)CX(2)NX(4)C, CXXC motif and 11 conserved cysteines. Phylogenetic analysis showed that AmphiGILT and sea urchin GILT clubbed together and positioned at the base of vertebrate GILT clade, suggesting that both AmphiGILT and sea urchin GILT might share some characteristics of the archetype of vertebrate GILT proteins. The genomic DNA sequence of B. floridae contains seven exons and six introns, which is similar to vertebrate GILT exon-intron organization. AmphiGILT was expressed in a tissue-specific manner with the most abundant mRNA in the digestive system including hepatic caecum and hind-gut. It was also found that mammalian IFN-gamma only exerted a slight effect on the expression of GILT gene in amphioxus, forming a contrast to the marked induction of human and mouse GILT expression by IFN-gamma. Taken the absence of the adaptive immune system including MHC class II molecules and lymphocytes into consideration, these results suggest that AmphiGILT is highly likely to play a role in the innate immune responses in amphioxus.

Traditional Mongolian medicine Eerdun Wurile improves stroke recovery through regulation of gene expression in rat brain.

ETHNOPHARMACOLOGICAL RELEVANCE: Eerdun Wurile (EW) is one of the key Mongolian medicines for treatment of neurological and cardiological disorders. EW is ranked most regularly used Mongolian medicine in clinic. Components of EW which mainly originate from natural products are well defined and are unique to Mongolian medicine. AIM OF THE STUDY: Although the recipe of EW contains known neuroactive chemicals originated from plants, its mechanism of action has never been elucidated at molecular level. The objective of the present study is to explore the mechanism of neuroregenerative activity of EW by focusing on the regulation of gene expression in the brain of rat model of stroke. MATERIALS AND METHODS: Rat middle cerebral artery occlusion (MCAO) models were treated with EW for 15 days. Then, total RNAs from the cerebral cortex of rat MCAO models treated with either EW or control (saline) were extracted and analyzed by transcriptome sequencing. Differentially expressed genes were analyzed for their functions during the recovery of ischemic stroke. The expression level of significantly differentially expressed genes such as growth factors, microglia markers and secretive enzymes in the lesion was further validated by RT-qPCR and immunohistochemistry. RESULTS: Previously identified neuroactive compounds, such as geniposide (Yu et al., 2009), myristicin (Shin et al., 1988), costunolide (Okugawa et al., 1996), toosendanin (Shi and Chen, 1999) were detected in EW formulation. Bederson scale indicated that the treatment of rat MCAO models with EW showed significantly lowered neurological deficits (p < 0.01). The regional cerebral blood circulation was also remarkably higher in rat MCAO models treated with EW compared to the control group. A total of 186 genes were upregulated in the lesion of rat MCAO models treated with EW compared to control group. Among them, growth factors such as Igf1 (p < 0.05), Igf2 (p < 0.01), Grn (p < 0.01) were significantly upregulated in brain after treatment of rat MCAO models with EW. Meanwhile, greatly enhanced expression of microglia markers, as well as complementary components and secretive proteases were also detected. CONCLUSION: Our data collectively indicated that EW enhances expression of growth factors including Igf1 and Igf2 in neurons and microglia, and may stimulate microglia polarization in the brain. The consequences of such activity include stimulation of neuron growth, hydrolysis and clearance of cell debris at the lesion, as well as the angiogenesis.

The Critical Role of Induced CD4+ FoxP3+ Regulatory Cells in Suppression of Interleukin-17 Production and Attenuation of Mouse Orthotopic Lung Allograft Rejection.

BACKGROUND: Lung transplantation is the only definitive therapy for many forms of end-stage lung disease. Studies have demonstrated the critical role of interleukin (IL)-17 in the development of lung rejection. Regulatory T cells (Tregs) are essential for the establishment and maintenance of immune tolerance. METHODS: We established mouse orthotopic lung transplantation models to investigate the importance of IL-17 and IL-17-producing cell types in acute lung allograft rejection and the efficacy of the adoptive transfer of induced Tregs (iTregs) in attenuating pathologic lesions of lung allografts. RESULTS: We found that the IL-17 produced by Th17 cells and γδ T cells might make the primary contributions to the progression of acute lung allograft rejection. Interleukin-17 deficiency decreased lung allograft lesions. Exogenous iTregs maintained their FoxP3 expression levels in lung allograft recipients. Induced Tregs therapy downregulated the expressions of Th17 and IL-17 γδ T cells and increased IL-10 production in the mouse orthotopic lung transplantation models. Moreover, the adoptive transfer of iTregs prolonged the survivals of the lung allografts and attenuated the progression of acute rejection. CONCLUSION: These data suggested that the adoptive transfer of iTregs could suppress the Th17 cells and IL-17 γδ cells of the recipients, decrease the expression of IL-17, and attenuate the pathology of acute lung allograft rejection. Exogenous iTregs upregulated immunosuppressive factors, such as IL-10 and suppressed IL-17-producing cells, which was one of the pathways to play a role in protecting lung allografts.

Ventilator-associated pneumonia in patients with cerebral hemorrhage: Impact on mortality and microbiological characterization / Neumonía asociada a ventilación mecánica en pacientes con hemorragia cerebral: mortalidad y caracterización microbiana

Ventilator-associated pneumonia (VAP) is a major complication among critically ill patients who depend on mechanical ventilation. Few reports have focused on intracerebral hemorrhage patients with VAP. Our main objective was to investigate the bacteria distribution characteristics and the impact of ventilator-associated pneumonia mortality in critical cerebral hemorrhage patients. This retrospective study included 89 cases of cerebral hemorrhage patients with VAP admitted to the ICU of Huashan Hospital. We used the chi-square test to compare qualitative variables and Student's t-test to compare means between groups of normally distributed quantitative variables. Multiple logistic regression analysis was used to assess mortality-independent predictors in the ICU. A total of 42% patients with cerebral hemorrhage were diagnosed with VAP in the ICU during the study period, and the mortality rate was 18%. Acinetobacter baumannii (n=58), Klebsiella pneumoniae (n=52), and Pseudomonas aeruginosa (n=21) were the most common pathogenic bacteria. Blood volume >30ml, tracheal ventilation mode and head of bed elevation were independent factors associated with increased mortality. Glasgow Coma Scale (GCS), Acute Physiology and Chronic Health Evaluation II (APACHE II) score and the time from bleeding to intubation were other potentially important factors. While the number of infecting bacteria may not be directly related to death, it can increase antibiotic consumption and length of intensive care unit (ICU) stays. Blood volume >30ml, tracheal ventilation mode and head of bed elevation were directly related to the death of critical cerebral hemorrhage patients with ventilator-associated pneumonia

La neumonía asociada a ventilación mecánica (NAV) es una complicación mayor entre los pacientes críticos que dependen de la ventilación mecánica. Pocos artículos se han centrado en los pacientes de hemorragia cerebral con NAV. Nuestro objetivo principal fue investigar las características de la distribución bacteriana y el impacto de la mortalidad de la neumonía asociada a ventilación mecánica en pacientes críticos de hemorragia cerebral. Este estudio retrospectivo incluyó 89 casos de pacientes de hemorragia cerebral con NAV ingresados en la unidad de cuidados intensivos (UCI) del Hospital Huashan. Utilizamos la prueba χ2 para comparar las variables cualitativas, y la t de Student para comparar las medias entre grupos de las variables cuantitativas normalmente distribuidas. Utilizamos análisis de regresión logística múltiple para valorar los factores predictivos independientes de la mortalidad en la UCI. Diagnosticamos NAV en el 42% de los pacientes con hemorragia en la UCI durante el periodo de estudio, y la tasa de mortalidad fue del 18%. Acinetobacter baumannii (n=58), Klebsiella pneumoniae (n=52) and Pseudomonas aeruginosa (n=21) fueron las bacterias patogénicas más comunes. El volumen sanguíneo >30ml, el modo de ventilación traqueal y la inclinación del cabecero de la cama fueron factores independientes asociados al incremento de la mortalidad. La Glasgow Coma Scale (GCS) y el Acute Physiology and Chronic Health EvaluationII (APACHE II), así como el tiempo transcurrido entre el sangrado y la intubación, fueron otros factores potencialmente importantes. A pesar de que el número de bacterias infecciosas puede no estar directamente relacionado con la muerte, puede incrementar el consumo de antibióticos y la duración de la estancia en la UCI. El volumen sanguíneo >30ml, el modo de ventilación traqueal y la inclinación del cabecero de la cama guardaron una relación directa con la muerte de los pacientes críticos de hemorragia cerebral con neumonía asociada a ventilación mecánica

Identification, expression, function and localization of a DUF985 domain-containing hypothetical gene from amphioxus Branchiostoma belcheri.

The progress in genome sequencing has led to an increasing submission of uncharacterized hypothetical genes with the domain of unknown function, DUF985, in GenBank, and none of these genes is related to a known protein. We therefore underwent an experimental study to identify the function of a DUF985 domain-containing hypothetical gene BbDUF985 (GenBank Accession No. AY273818) isolated from amphioxus Branchiostoma belcheri (B. belcheri). BbDUF985 was successfully expressed in both prokaryotic and eukaryotic systems, and its recombinant proteins expressed in both systems definitely exhibited an activity of phosphoglucose isomerase (PGI). Both tissue-section in situ hybridization and immunohistochemistry demonstrated that BbDUF985 was expressed in a tissue-specific manner, with most abundant levels in the hepatic caecum and ovary. In CHO cells transfected with the expression plasmid pEGFP-N1/BbDUF985, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that BbDUF985 is a cytosolic protein. In contrast, Western blotting indicated that BbDUF985 was also present in amphioxus humoral fluids, suggesting that it exists as a secreted protein as well. Our study provided a framework for further understanding the biochemical properties and physiological function of DUF985-containing hypothetical proteins in other species.

Analysis of risk factors related with carotid atherosclerotic arteriosclerosis in a Chinese young and middle-aged population who received health checkup / 中华健康管理学杂志

Objective To investigate the relationship between carotid artery atherosclerosis and risk factor in a young and middle-aged population in China who received health checkup.Methods A total of 762 cases of young and middle-aged adult were recruited between April and October in 2013.These subjects had no hypertension,diabetes,coronary heart disease,cerebrovascular disease,and connective tissue disease,etc.Their Blood pressure,serum glucose (Glu),total cholesterol (TC),triglyceride (TG),and low density lipoprotein cholesterol (LDL-C) were measured and carotid artery ultrasonography was performed.Of the 762 cases,589 were male,173 were female,mean age was (46 ± 9) years.For blood pressure,blood lipids,the t test was performed to detect the difference between arteriosclerosis group and non arteriosclerosis group in different age groups.Logistic regression was performed to reveal the risk factors with carotid arteriosclerosis.Results Of the 762 subjects,205 had carotid artery atherosclerosis; 30.2％ (178/589) of them were men and 15.6％(27/173) of them were women,with a significant difference between two groups,(x2=14.522,P=0.000).In the younger than 40 years old group,the diastolic blood pressure has statistical significance between carotid arteriosclerosis and non carotid arteriosclerosis group (t=-2.789,P＜ 0.05); blood sugar had statistically significant difference between the two groups (t value was-2.256,-2.930,-2.174,respectivly,P＜0.05).Multiple regression analysis revealed that the independent risk factors for carotid artery arteriosclerosis were age,sex,DBP,and Glu.The regression coefficients were 0.993,0.711,0.047,0.334 seperately.The ROC curve of age was analyzed,the area under the ROC curve was 0.731,cut-off was 51 years old.Conclusions Routine carotid artery ultrasound should be performed for older than 50 healthy middle-aged men,to determine whether arteriosclerosis exists,especially for people with abnormal blood glucose or elevated DBP.