Analysis of the Differential Expression and Antiviral Activity of Porcine Interferon-α In Vitro.

Porcine interferon α (poIFN-α) is a crucial cytokine that can prevent and treat viral infections. Seventeen functional porcine IFN-α subtypes were found in the porcine genome. In this study, multiple sequence alignment was performed to analyze IFN-α protein structure and function. Phylogenetic tree analysis of the poIFN gene family defined the evolutionary relationship of various subtypes. PoIFN-αs, including poIFN-α1-17, were expressed in an Escherichia coli expression system. The antiviral activities of these IFN-α proteins against vesicular stomatitis virus (VSV) and pseudorabies virus (PRV) were examined in PK-15 cells. We found that the antiviral activity of different poIFN-α molecules greatly differed as follows: the poIFN-α14 and 17 subtypes had the greatest antiviral activities against VSV and PRV in PK-15 cells, poIFN-α1, 2, 3, and 8 exhibited lower biological activities, and poIFN-α4, 5, 6, 7, 9, 10, 11, 12, 13, and 16 had minimal or no effect in the tested target cell‒virus systems. Moreover, our studies demonstrated that the antiviral activity of IFN-α was positively correlated with the induction of IFN-stimulated genes, such as 2'-5' oligoadenylate synthetase 1 (OSA1), interferon-stimulated gene 15 (ISG15), myxoma resistance protein 1 (Mx1), and protein kinase R (PKR). Thus, our experimental results provide important information about the antiviral functions and mechanism of poIFN-α.

Fluorescent Probe Combined with Photoelectric Analysis Technology for Detection of Escherichia coli.

Food safety is facing great challenges in preventing foodborne diseases caused by pathogenic pollution, especially in resource-limited areas. The rapid detection technique of microorganisms, such as immunological methods and molecular biological methods, plays a crucial key in timely bioanalysis and disease treatment strategies. However, it is difficult for these methods to simultaneously meet the criteria of simple operation, high specificity, and sensitivity, as well as low cost. Coconut water is known as the "water of life" in Hainan. It is a refreshing and nutritious beverage which is widely consumed due to its beneficial properties to health. Coconut water processing is an important pillar industry in Hainan. The detection of pathogenic microorganisms, such as Escherichia coli, in coconut water has become an important factor which has restricted the upgrading and development of this industry. Based on the needs of industrial development, we developed a microbial photoelectric detection system which was composed of a fluorescent probe detection reagent and a photoelectric sensor detection device. This system combined microbial enzyme targets, selective fluorescent substrate metabolism characteristics, and a photoelectric sensor signal transduction mechanism, which produce a strong signal with a high signal-to-noise ratio. The microbial detection system developed here has a simple structure, simple and convenient operation, short detecting time (≥2 h), and high sensitivity (1 CFU/mL). This system may also enable early warning and monitoring programs for other pathogenic microorganisms in order to promote the overall competitiveness of the Hainan coconut water industry.

Explorations on Thermodynamic and Kinetic Performances of Various Cationic Exchange Durations for Synthetic Clinoptilolite.

Various cation-exchanged clinoptilolites (M-CPs, M = Li+, Cs+, Ca2+, Sr2+) were prepared, and their exchanged thermodynamic (and kinetic) properties and adsorption performances for CH4, N2, and CO2 were investigated. The results demonstrated that the relative crystallinity of M-CPS decreased with the increase of exchange times. Their chemisorbed water weight loss gradually increased with the increasing exchange times, except that of Cs-x-CP. The ΔrGmθ values of exchange process of Li+, Cs+, Ca2+, or Sr2 presented the increased trend with the enhanced exchange times, but they decreased as the temperature increased. The negative ΔrGmθ values and the positive ΔrHmθ and ΔrSmθ values suggested that the exchanged procedure belonged to spontaneous, endothermic, and entropy-increasing behaviors; their kinetic performances followed a pseudo-second-order model. However, the calculated Ea values of exchange process showed the increased tendencies with the enhanced exchange times, indicating that the exchange process became more difficult. Finally, the preliminary adsorption results indicated that the maximum adsorption amount at 273 K and 1 bar was 0.51 mmol/g of CH4 and 0.38 mmol/g of N2 by (Na, K)-CP, and 2.32 mmol/g of CO2 by Li-6-CP.

AMP-activated kinase regulates porcine reproductive and respiratory syndrome virus infection in vitro.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen in the pig industry worldwide. Many viruses manipulate their cellular metabolism to replicate themselves and cause infection. A conserved cellular energy sensor, 5'-AMP-activated protein kinase (AMPK), maintains cellular energy homeostasis. We found that PRRSV infection caused significant AMPK activation in a time-dependent manner via the ROS-calcium/calmodulin-dependent protein kinase-2 pathway. RNA interference-mediated AMPK knockdown could increase PRRSV replication in MARC-145 cells, suggesting that AMPK contributed to PRRSV infection regulation. Moreover, investigation of the effect of AMPK activity on PRRSV replication showed that PRRSV replication could be suppressed by the pharmacological agonists 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside and A769662. Conversely, an AMPK inhibitor, compound C, markedly enhanced PRRSV infection. Furthermore, the AMPK agonist A769662 was found to exert no effect on PRRSV entry, assembly, and release, suggesting that A769662 may hinder the PRRSV genome replication in MARC-145 cells. In conclusion, AMPK may be a promising antiviral drug target against PRRSV infection.

Quantitative Proteomic Analysis of Global Protein Acetylation in PRRSV-Infected Pulmonary Alveolar Macrophages.

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), is a serious viral disease affecting global swine industry. Due to the lack of effective vaccines, new antiviral strategies to compensate for the inefficacy of available vaccines are urgently required. Lysine acetylation, as an important post-translational modification during infection, plays a key regulatory role in host antiviral responses. In this study, the global acetylome is profiled using acetylation specific antibody-based enrichment and tandem mass tag label high-affinity purification liquid chromatography-mass spectrometry in PRRSV-infected pulmonary alveolar macrophages (PAMs). As a result, 3731 lysine acetylation sites on 1421 cellular proteins are identified. Bioinformatics analysis of the different acetylated proteins revealed their involvement in various biological processes, including the host immune response and energy metabolism. These findings will contribute to the understanding of PRRSV pathogenesis and identify new cellular targets for anti-PPRSV therapeutics.

Aerosols in an arid environment: The role of aerosol water content, particulate acidity, precursors, and relative humidity on secondary inorganic aerosols.

Meteorological conditions, gas-phase precursors, and aerosol acidity (pH) can influence the formation of secondary inorganic aerosols (SIA) in fine particulate matter (PM2.5). Most works related to the influence of pH and gas-phase precursors on SIA have been laboratory research, but field observation research is very scarce, especially in arid environments. The relationship among SIA, pH, gas-phase precursors, and meteorological conditions are investigated in Hohhot, a major city in China with an arid environment. Secondary inorganic species, e.g., SO42-, NO3-, were typically found at low levels, reflecting the low level of secondary aerosol. It is interesting to note that the level of SO2 in Hohhot was higher than in other cities while SO42- was relatively lower than in other cities. Multiple receptor models were used to explore the contributions to the SIA and quantify the source impacts on the SIA. Annual average aerosol pH in Hohhot was 5.6 (range 1.1-8.4) which was estimated by a thermodynamic equilibrium model. Additionally, a statistical method was used to evaluate the influence of SIA sources on ambient aerosol concentrations. Aerosol water content and particulate acidity were found to be positively associated with secondary SO42-, while NO2 and RH had a significant impact on secondary NO3- in an arid atmosphere. The findings explain the relationship between gaseous precursors, relative humidity, aerosol pH and temperature in the arid city of Hohhot.

Embryonic exposure to ethanol increases the susceptibility of larval zebrafish to chemically induced seizures.

Prenatal ethanol exposure is known to cause neurodevelopmental disorders. While high prevalence of epilepsy is observed among the children whose mothers abused alcohol during pregnancy, the results from animal studies are conflicting. Here, we investigated whether embryonic exposure to ethanol can increase the susceptibility to pentylenetetrazole (PTZ)-induced seizures in larval zebrafish. Embryos at 3 hours post-fertilization (hpf) were exposed to ethanol at the concentrations ranging from 0.25% to 1% for 21 hours. Control and ethanol-exposed larvae were challenged with PTZ at 7 days post-fertilization (dpf) at the concentrations of 2.5, 5 or 15 mM. The seizure behavior of larvae was recorded and analyzed using EthoVision XT 11. We found that embryonic ethanol exposure increased the percentage of larvae exhibiting typical stage II and III seizure and resulted in a significant reduction in stage I, II and III seizure latency in an ethanol concentration-dependent manner. Embryonic exposure to ethanol also significantly increased the severity of PTZ-induced seizures in larvae, as demonstrated by increased total distance traveled and the duration of mobility. This is the first demonstration that ethanol exposure during early embryonic stage can reduce the threshold for chemically induced seizures and increase the severity of seizure behavior in larval fish.

Sirolimus alternative to blood transfusion as a life saver in blue rubber bleb nevus syndrome: A case report.

RATIONALE: Blue rubber bleb nevus syndrome (BRBNS) is a rare disease characterized by multiple venous malformations. The gastrointestinal bleeding and secondary iron deficiency anemia are the most common complications. There are currently no effective treatments for BRBNS. Here, we report a case of successful treatment with a small dose of sirolimus of a BRBN patient with a de novo gene mutation. PATIENT CONCERNS: A 12-year-old female was admitted to our hospital with multiple hemangiomas for 12 years. The patient often displayed melena; she recently received transfusion of 2 units of red blood cells once every 2 weeks. Multiple fist-sized hemangiomas were piled up on both sides and back of the neck, and were also noted on the arms, legs, chest, back, and on the tip of the tongue. The laboratory findings demonstrated severe anemia. Blood sample sequencing detected a heterozygous de novo mutation c.2545C > Tin the TEK gene. DIAGNOSES: Based on these findings, final diagnosis of Blue rubber bleb nevus syndrome (BRBNS) was made. INTERVENTIONS: After the diagnosis, low-dose sirolimus was orally administered. OUTCOMES: The patient's hemoglobin was increased after treatment with sirolimus for 1 month. Since the initial treatment with sirolimus, she had not received any blood transfusions. The skin and mucosal hemangioma decreased significantly, and new digestive tract hemorrhage, muscle hematoma, or adverse drug reactions were not observed. LESSONS: we report a case of a mutation in exon 15 of the TEK gene leading to BRBN. It was successfully treated with a small dose of sirolimus as an alternative to blood transfusion in order to save the of BRBN patient's life.

The role of T-type calcium channel genes in absence seizures.

The thalamic relay neurons, reticular thalamic nucleus, and neocortical pyramidal cells form a circuit that sustains oscillatory burst firing, and is regarded as the underlying mechanism of absence seizures. T-type calcium channels play a key role in this circuit. Here, we review the role of T-type calcium channel genes in the development of absence seizures, and emphasize gain or loss of function mutations, and other variations that alter both quantity and quality of transcripts, and methylation status of isoforms of T-type calcium channel proteins might be of equal importance in understanding the pathological mechanism of absence seizures.

[Bioimmunological characteristics of mature or immature murine dendritic cells].

OBJECTIVE: To explore the biological characteristics of immature and mature murine bone marrow-derived dendritic cells (DCs). METHODS: The murine bone marrow cells were cultured and induced in vitro into immature DCs (imDCs). Then ImDCs were incubated with TNF (tumor necrosis factor)-α to obtain mature DCs (mDCs). The DCs were purified by murine CD11c microbeads. The morphologies of DCs were observed by electron microscopy (EM). Such surface markers as MHC-II, CD80 and CD86 were tested by flow cytometry (FCM). The proliferation capacity of allogeneic T cells stimulated by DCs was examined by Cell Counting Kit-8 (CCK-8) in mixed lymphocyte reaction (MLR). The expressions of cell factors in mRNA of DCs were tested by real-time polymerase chain reaction (QPCR). The cytokines in supernatant were measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: As compared with mDCs, fewer, shorter spines and more phagocytic vesicles and lysosome were observed in imDCs under EM. The expressions of cell surface molecules in imDCs were significantly lower than those of mDC by FCM, [MHC-II (27.2%) vs (97.7%); CD80 (27.6%) vs (97.2%); CD86 (29.5%) vs (96.4%)]. In MLR, the capacity of same-reaction ratio imDCs group for stimulating the proliferation of T-cells was remarkably lower than that of mDCs group,[1:5 (1.63 ± 0.04) vs (2.21 ± 0.09); 1:10 (1.50 ± 0.08) vs (1.92 ± 0.02); 1:20 (1.28 ± 0.07) vs (1.64 ± 0.01); 1:40 (1.19 ± 0.04) vs (1.45 ± 0.06), P < 0.01]. In imDCs, the relative mRNA expressions of IL12p35 (0.66 ± 0.13), IL12p40 (0.57 ± 0.10) and IFN-γ (0.74 ± 0.08) were lower than those of mDCs (1.00 ± 0.00), (P < 0.05), but the expression of TGF-ß (1.35 ± 0.09) was higher than that of mDCs by QPCR (1.00 ± 0.00), (P < 0.05). The expressions of IL12p70 and IFN-γ in supernatants from imDCs were lower than those of mDCs [IL12p70: (6 ± 4) vs (120 ± 22); IFN-γ: (56 ± 15) vs (90 ± 15), P < 0.05] while the expression of TGF (transforming growth factor)-ß was higher than that of mDCs by ELISA [TGF-ß: (176 ± 23) vs (55 ± 18), P < 0.05]. CONCLUSION: ImDCs can induce anergic T cells and immune tolerance through a down-regulation of co-stimulatory molecules, MHC-II and Th1 cytokines and an up-regulation of Th2 cytokine (TGF-ß). And MDCs may activate naive T cells and stimulate immune responses by an up-regulation of co-stimulatory molecules, MHC-II and Th1 cytokines.

[Changes of body weight and galanin in epileptic children treated with topiramate].

OBJECTIVE: Topiramate is a new broad-spectrum anti-epileptic drug. Decreased body weight and appetite are common side effects of topiramate. The side effect affects the growth and development in children greatly. Little is known about the mechanisms of topiramate-induced weight loss and decreased appetite in children with epilepsy in China and abroad. galanin is one of factors that affect appetite. It is a neuroendocrine peptide and play an important role in the control of appetite and body weight in the mechanism of hormone release. The purpose of this study was to explore the mechanism of topiramate-induced weight loss in children with epilepsy and the relation of weight loss with change of galanin, thereby to provide evidences for improvement of quality of life, compliance to treatment and reduce side effects of growth and development in children with epilepsy. METHODS: Totally 61 patients with especial epilepsy were enrolled into this study and the disease was defined by clinical manifestations and electroencephalography (EEG). Among them 32 cases had generalized seizures and 29 had local seizures. Sixteen normal children were enrolled as control group. The patients' age ranged from 0.5 to 14 (4.76 +/- 4.05) years and the patients were instructed to take 0.5 - 1 mg/kg of topiramate per day, with 0.5 - 1 mg/kg every 3 - 5 d increased to maximum of 3 - 8 mg/kg per day. Patients continued receiving the doses for 4 months. All patients' serum galanin levels and body height and weight and hepatic function were detected before and after antiepileptic drugs treatment. The galanin was detected by using radioimmunoassay. RESULTS: After treatment with topiramate (61 cases) for 4 months, plasma galanin [(22.01 +/- 8.12) pg/ml] declined as compared with baseline [(26.56 +/- 9.35) pg/ml, t = 2.85, P < 0.01] in children with epilepsy. Twenty-two of 61 patients lost weight, their plasma galanin concentration was significantly lower [(26.51 +/- 10.00) pg/ml vs. (20.45 +/- 8.09) pg/ml, t = 2.91, P < 0.01], but there was no significant change in the weight-gained patients (39/61) and control group (n = 16). In children with epilepsy, the mean value of body weight decreased as compared with the pre-treatment values, but the difference was not significant; however, the body-mass index (BMI) was significantly lower than that obtained before treatment (t = 8.628, P < 0.01). Eighteen of 22 patients who lost weight had decreased appetite, but only five of 39 patients who gained weight showed decreased appetite (chi(2) = 28.50, P < 0.001). The mean value of plasma galanin declined after treatment in patients (23 cases) with decreased appetite [(18.35 +/- 7.80) pg/ml vs. (27.28 +/- 6.90) pg/ml, t = 4.84, P < 0.001]; while plasma galanin did not change significantly after treatment in patients (38 cases) without decreased appetite [(24.23 +/- 7.66) pg/ml vs. (26.12 +/- 5.49) pg/ml, t = 1.04, P > 0.05]. CONCLUSION: Topiramate treatment may lower the body weight and reduce appetite in part of children with epilepsy which may be mediated by the reduced plasma galanin level.