Pathological variations and immune response in Channa argus infected with pathogenic Nocardia seriolae strain.

Nocardia seriolae pathogen causes chronic granulomatous disease, reportedly affecting over 40 species of marine and freshwater cultured fish. Hence, research is required to address and eliminate this significant threat to the aquaculture industry. In this respect, a reliable and reproducible infection model needs to be established to better understand the biology of this pathogen and its interactions with the host during infection, as well as to develop new vaccines or other effective treatment methods. In this study, we examined the pathogenicity of the pathogen and the immune response of snakehead (Channa argus) juvenile to N. seriolae using a range of methods and analyses, including pathogen isolation and identification, histopathology, Kaplan-Meier survival curve analysis, and determination of the median lethal dose (LD50) and cytokine expression. We have preliminarily established a N. seriolae - C. argus model. According to our morphological and phylogenetic analysis data, the isolated strain was identified as N. seriolae and named NSE01. Eighteen days post-infection of healthy juvenile C. argus with N. seriolae NSE01, the mortality rate in all four experimental groups (intraperitoneally injected with 1 × 105 CFU/mL - 1 × 108 CFU/mL of bacterial suspension) (n = 120) was 100 %. The LD50 of N. seriolae NSE01 for juvenile C. argus was determined to be 1.13 × 106 CFU/fish. Infected juvenile C. argus had significant pathological changes, including visceral tissue swelling, hemorrhage, and the presence of numerous nodules of varying sizes in multiple tissues. Further histopathological examination revealed typical systemic granuloma formation. Additionally, following infection with N. seriolae NSE01, the gene expression of important cytokines, such as Toll-like receptor genes TLR2, TLR13, interleukin-1 receptor genes IL1R1, IL1R2, and interferon regulatory factor IRF2 were significantly upregulated in different tissues, indicating their potential involvement in the host immune response and regulation against N. seriolae. In conclusion, juvenile C. argus can serve as a suitable model for N. seriolae infection. The establishment of this animal model will facilitate the study of the pathogenesis of nocardiosis and the development of vaccines.

Noninvasive detection of diabetes mellitus based on skin fluorescence and diffuse reflectance spectroscopy.

There is an urgent need for a mass population screening tool for diabetes. Skin tissue contains a large number of endogenous fluorophores and physiological parameter markers related to diabetes. We built an excitation-emission spectrum measurement system with the excited light sources of 365, 395, 415, 430, and 455 nm to extract skin characteristics. The modeling experiment was carried out to design and verify the accuracy of the recovery of tissue intrinsic discrete three-dimensional fluorescence spectrum. Blood oxygen modeling experiment results indicated the accuracy of the physiological parameter extraction algorithm based on the diffuse reflectance spectrum. A community population cohort study was carried out. The tissue-reduced scattering coefficient and scattering power of the diabetes were significantly higher than normal control groups. The Gaussian multi-peak fitting was performed on each excitation-emission spectrum of the subject. A total of 63 fluorescence features containing information such as Gaussian spectral curve intensity, central wavelength position, and variance were obtained from each person. Logistic regression was used to construct the diabetes screening model. The results showed that the area under the receiver operating characteristic curve of the model for predicting diabetes was 0.816, indicating a high diagnostic value. As a rapid and non-invasive detection method, it is expected to have high clinical value.

Effect of Temperature Cycling Pretreatment on the Thermal Stability of Sm2(Co, Fe, Zr, Cu)17 Magnets in the Mild Temperature Range.

The irredeemable magnetic losses of Sm(Co, Fe, Zr, Cu)7.8 permanent magnets caused by oxidation are very important for their practical application. In this work, the simulated results with R2 ≥ 98% based on the data of the temperature cycling test and the long-term isothermal test for the original samples confirmed that the magnetic flux losses reached 9.38% after the 5000th cycle in range R.T.-300 °C, and 7.15% after oxidated at 180 °C for 10 years, respectively. Demagnetization curves showed that the low-temperature oxidation mainly led to the remanence attenuation, while the coercivity remained relatively stable. SEM observation and EDS analysis revealed that an oxide outer layer with a thickness of 1.96 µm was formed on the surface of the original sample at 180 °C for 180 days, in which there was no enrichment or precipitation of metal elements. However, once a Cu, O-rich outer layer with a thickness of 0.72 µm was grown by using a temperature cycling from -50-250 °C for three cycles, the attenuation of magnetic properties could be inhibited under the low-temperature oxidation. This work suggested that the magnetic attenuation of Sm2Co17-type permanent magnets in the low-temperature field could not be ignored, and provided a simple method to suppress this attenuation of magnetic properties below 300 °C.

Identification of key pathways and genes in the progression of silicosis based on WGCNA.

Silicosis, induced by inhaling silica particles in workplaces, is one of the most common occupational diseases. The prognosis of silicosis and its consequent fibrosis is extremely poor due to limited treatment modalities and lack of understanding of the disease mechanisms. In this study, a Wistar rat model for silicosis fibrosis was established by intratracheal instillation of silica (0, 50, 100 and 200 mg/mL, 1 mL) with the evidence of Hematoxylin and Eosin (HE) and Masson staining and the expressions of inflammatory and fibrotic proteins of rats' lung tissues. RNA of lung tissues of rats exposed to 200 mg/mL silica particles and normal saline for 14 d and 28 d was extracted and sequenced to detect differentially expressed genes (DEGs) and to identify silicosis fibrosis-associated modules and hub genes by Weighted gene co-expression network analysis (WGCNA). Predictions of gene functions and signaling pathways were conducted using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. In this study, it has been demonstrated the promising role of the Hippo signaling pathway in silicosis fibrosis, which will be conducive to elucidating the specific mechanism of pulmonary fibrosis induced by silica and to determining molecular initiating event (MIE) and adverse outcome pathway (AOP) of silicosis fibrosis.

An innovative S-scheme AgCl/MIL-100(Fe) heterojunction for visible-light-driven degradation of sulfamethazine and mechanism insight.

The development of high efficient photocatalysts for antibiotics contamination in water remains a severe challenge. In this study, a novel step-scheme (S-scheme) photocatalytic heterojunction nanocomposites were fabricated from integrating AgCl nanoparticles on the MIL-100(Fe) octahedron surface through facile multi-stage stirring strategy. The S-scheme heterojunction structure in AgCl/MIL-100(Fe) (AM) nanocomposite provided a more rational utilization of electrons (e-) and holes (h+), accelerated the carrier transport at the junction interface, and enhanced the overall photocatalytic performance of nanomaterials. The visible-light-driven photocatalysts were used to degrade sulfamethazine (SMZ) which attained a high removal efficiency (99.9%). The reaction mechanisms of SMZ degradation in the AM photocatalytic system were explored by electron spin resonance (ESR) and active species capture experiments, which superoxide radical (•O2-), hydroxyl radical (•OH), and h+ performed as major roles. More importantly, the SMZ degradation pathway and toxicity assessment were proposed. There were four main pathways of SMZ degradation, including the processes of oxidation, hydroxylation, denitrification, and desulfonation. The toxicity of the final products in each pathway was lower than that of the parent according to the toxicity evaluation results. Therefore, this work might provide new insights into the environmentally-friendly photocatalytic processes of S-scheme AM nanocomposites for the efficient degradation of antibiotics pollutants.

A quadruple protection procedure for resuming pig production in small-scale ASFV-positive farms in China.

African swine fever (ASF) outbreak has caused serious economic losses in Asia since 2018. As ASF is a new emerging disease, many farmers hesitate to raise pigs before biosafety procedures were evaluated to be effective. To support small-scale farms in resuming pig production, a comprehensive procedure, called the quadruple protection procedure (QPP), was tested in 35 small farms which had been confirmed with African swine fever virus (ASFV). The QPP takes care of the farms' construction, environmental disinfection, regular immunization, and feed quality. Qualified daily management was supplemented as well. During a one-year survey four disinfectants and one piece of equipment were used in higher frequency. A 7- or 15-day empty period after the disinfection was suitable when it was combined with the rest of the protection measures from QPP. Totally 18,730 porkers and 3,006 sows were healthy by the end of the study with percentage of 100 and 98.8, respectively, indicating that QPP could protect pigs in small-scale farms from pathogens within China. This study developed an effective protective procedure system for small-scale farms to produce pigs under the risk of ASF outbreak.

Mirror-like Bright Al-Mn Coatings Electrodeposition from 1-Ethyl-3 Methylimidazolium Chloride-AlCl3-MnCl2 Ionic Liquids with Pyridine Derivatives.

The effects of three pyridine derivative additives, 4-hydroxypyridine, 4-picolinic acid, and 4-cyanopyridine, on Al-Mn coatings were investigated in 1-ethyl-3-methylimidazolium chloride-AlCl3-MnCl2 (EMIC-AlCl3-MnCl2) ionic liquids. The smooth mirror-like bright Al-Mn coatings were obtained only in the EMIC-AlCl3-MnCl2 ionic liquids containing 4-cyanopyridine, while the matte Al-Mn coatings were electrodeposited from EMIC-AlCl3-MnCl2 without additives or containing either 4-hydroxypyridine or 4-picolinic acid. The scanning electron microscope and X-ray diffraction showed that the bright Al-Mn coatings consisted of nanocrystals and had a strong (200) preferential orientation, while the particle size of matte Al-Mn coatings were within the micron range. The brightening mechanism of 4-cyanopyridine is due to it being adsorbed onto the cathode to produce the combined effect of (1) generating an overpotential to promote Al-Mn nucleation; (2) inhibiting the growth of the deposited nuclei and enabling them grow preferentially, making the coating composed of nanocrystals and with a smooth surface. The brightening effect of 4-cyanopyridine on the Al-Mn coatings was far better than that of the 4-hydroxypyridine and the 4-picolinic acid. In addition, the bright Al-Mn coating was prepared in a bath with 6 mmol·L-1 4-cyanopyridine and displayed superior corrosion resistance relative to the matte coatings, which could be attributed to its unique nanocrystalline structure that increased the number of grain boundaries and accelerated the formation of the protective layer of the corrosion products.

Enhancing the antifouling and rejection properties of PVDF membrane by Ag3PO4-GO modification.

Ultrafiltration is an environmentally friendly water treatment technology, but membrane fouling significantly impacts membrane performance and service life. Photocatalytic modification of membrane is regarded as an effective way for membrane fouling control. In this study, graphite oxide (GO), Ag3PO4 and Ag3PO4-GO nanomaterials were applied in polyvinylidene fluoride (PVDF) ultrafiltration membranes modification, and the membranes was denoted as P-GO, P-AgP and P-AgP@GO, respectively. Filtration of humic acid (HA) at different operating conditions was adopted in evaluation of membrane performance. Among them, P-AgP@GO had the best permeation, rejection and antifouling performances, and could maintain excellent properties when operation conditions (HA concentration, operation pressure, pH and ionic strength) were changed. Furthermore, the effect of photocatalysis on the self-cleaning performance and its mechanism were revealed. The overall performance of P-AgP@GO could be enhanced by visible light irradiation, and extending the visible illumination time during the filtration was conducive to the reusability.

Development of a non-invasive method for skin cholesterol detection: pre-clinical assessment in atherosclerosis screening.

BACKGROUND: Establishing a high-accuracy and non-invasive method is essential for evaluating cardiovascular disease. Skin cholesterol is a novel marker for assessing the risk of atherosclerosis and can be used as an independent risk factor of early assessment of atherosclerotic risk. METHODS: We propose a non-invasive skin cholesterol detection method based on absorption spectroscopy. Detection reagents specifically bind to skin cholesterol and react with indicator to produce colored products, the skin cholesterol content can be obtained through absorption spectrum information on colored products detected by non-invasive technology. Gas chromatography is used to measure cholesterol extracted from the skin to verify the accuracy and reliability of the non-invasive test method. A total of 342 subjects were divided into normal group (n = 115), disease group (n = 110) and risk group (n = 117). All subjects underwent non-invasive skin cholesterol test. The diagnostic accuracy of the measured value was analyzed by receiver-operating characteristic (ROC) curve. RESULTS: The proposed method is able to identify porcine skin containing gradient concentration of cholesterol. The values measured by non-invasive detection method were significantly correlated with gas chromatography measured results (r = 0.9074, n = 73, p < 0.001). Bland-Altman bias was - 72.78 ± 20.03 with 95% limits of agreement - 112.05 to - 33.51, falling within the prespecified clinically non-significant range. We further evaluated the method of patients with atherosclerosis and risk population as well as normal group, patients and risk atherosclerosis group exhibited higher skin cholesterol content than normal group (all P < 0.001). The area under the ROC curve for distinguishing Normal/Disease group was 0.8642 (95% confidence interval, 0.8138 to 0.9146), meanwhile, the area under the ROC curve for distinguishing Normal/Risk group was 0.8534 (95% confidence interval, 0.8034 to 0.9034). CONCLUSIONS: The method demonstrated its capability of detecting different concentration of skin cholesterol. This non-invasive skin cholesterol detection system may potentially be used as a risk assessment tool for atherosclerosis screening, especially for a large population.

The Key Regulator of Necroptosis, RIP1 Kinase, Contributes to the Formation of Astrogliosis and Glial Scar in Ischemic Stroke.

Necroptosis initiation relies on the receptor-interacting protein 1 kinase (RIP1K). We recently reported that genetic and pharmacological inhibition of RIP1K produces protection against ischemic stroke-induced astrocytic injury. However, the role of RIP1K in ischemic stroke-induced formation of astrogliosis and glial scar remains unknown. Here, in a transient middle cerebral artery occlusion (tMCAO) rat model and an oxygen and glucose deprivation and reoxygenation (OGD/Re)-induced astrocytic injury model, we show that RIP1K was significantly elevated in the reactive astrocytes. Knockdown of RIP1K or delayed administration of RIP1K inhibitor Nec-1 down-regulated the glial scar markers, improved ischemic stroke-induced necrotic morphology and neurologic deficits, and reduced the volume of brain atrophy. Moreover, knockdown of RIP1K attenuated astrocytic cell death and proliferation and promoted neuronal axonal generation in a neuron and astrocyte co-culture system. Both vascular endothelial growth factor D (VEGF-D) and its receptor VEGFR-3 were elevated in the reactive astrocytes; simultaneously, VEGF-D was increased in the medium of astrocytes exposed to OGD/Re. Knockdown of RIP1K down-regulated VEGF-D gene and protein levels in the reactive astrocytes. Treatment with 400 ng/ml recombinant VEGF-D induced the formation of glial scar; conversely, the inhibitor of VEGFR-3 suppressed OGD/Re-induced glial scar formation. RIP3K and MLKL may be involved in glial scar formation. Taken together, these results suggest that RIP1K participates in the formation of astrogliosis and glial scar via impairment of normal astrocyte responses and enhancing the astrocytic VEGF-D/VEGFR-3 signaling pathways. Inhibition of RIP1K promotes the brain functional recovery partially via suppressing the formation of astrogliosis and glial scar.

KIF15 facilitates gastric cancer via enhancing proliferation, inhibiting apoptosis, and predict poor prognosis.

BACKGROUND: Kinesin superfamily proteins (KIFs) can transport membranous organelles and protein complexes in an ATP-dependent manner. Kinesin family member 15 (KIF15) is overexpressed in various cancers. However, the function of KIF15 in gastric cancer (GC) is still unclear. METHODS: GC patients' data from The Cancer Genome Atlas (TCGA) were analyzed by bioinformatics methods. The expression of KIF15 was examined in GC and paracarcinoma tissues from 41 patients to verify the analysis results. The relationship between KIF15 expression and clinical characteristics were also observed by bioinformatics methods. Kaplan-Meier survival analysis of 122 GC patients in our hospital was performed to explore the relationship between KIF15 expression levels and GC patients' prognosis. KIF15 was downregulated in GC cell lines AGS and SGC-7901 by transfecting a lentivirus-mediated shRNA plasmid targeting KIF15. In vitro, GC cell proliferation and apoptosis were detected by MTT assay, colony formation assay, and Annexin V-APC staining. In vivo, xenograft experiments were used to verify the in vitro results. Furthermore, Human Apoptosis Antibody Array kit was used to screen possible targets of KIF15 in GC cell lines. RESULTS: The bioinformatics results showed that KIF15 expression levels were higher in GC tissues than in normal tissues. IHC showed same results. High expression of KIF15 was statistical correlated with high age and early histologic stage. Kaplan-Meier curves indicated that high KIF15 expression predict poor prognosis in patients with GC. MTT assay and colony formation assay showed that KIF15 promote GC cell proliferation. Annexin V-APC staining found that KIF15 can inhibit GC cell apoptosis. Xenograft experiments reveal that downregulating KIF15 can inhibit GC tumor growth and promote GC apoptosis. Through detection of 43 anti-apoptotic proteins by the Human Apoptosis Antibody Array kit, it was confirmed that knocking down KIF15 can reduce seven anti-apoptotic proteins expression. CONCLUSIONS: Taken together, our study revealed a critical role for KIF15 to inhibit GC cell apoptosis and promote GC cell proliferation. KIF15 may decrease anti-apoptotic proteins expression by regulating apoptosis pathways. High expression of KIF15 predicts a poor prognosis in patients with GC. KIF15 might be a novel prognostic biomarker and a therapeutic target for GC.

Lessons learned from the 2019-nCoV epidemic on prevention of future infectious diseases.

Only a month after the outbreak of pneumonia caused by 2019-nCoV, more than forty-thousand people were infected. This put enormous pressure on the Chinese government, medical healthcare provider, and the general public, but also made the international community deeply nervous. On the 25th day after the outbreak, the Chinese government implemented strict traffic restrictions on the area where the 2019-nCoV had originated-Hubei province, whose capital city is Wuhan. Ten days later, the rate of increase of cases in Hubei showed a significant difference (p = 0.0001) compared with the total rate of increase in other provinces of China. These preliminary data suggest the effectiveness of a traffic restriction policy for this pandemic thus far. At the same time, solid financial support and improved research ability, along with network communication technology, also greatly facilitated the application of epidemic prevention measures. These measures were motivated by the need to provide effective treatment of patients, and involved consultation with three major groups in policy formulation-public health experts, the government, and the general public. It was also aided by media and information technology, as well as international cooperation. This experience will provide China and other countries with valuable lessons for quickly coordinating and coping with future public health emergencies.

Association of total oxidant status, total antioxidant status, and malondialdehyde and catalase levels with psoriasis: a systematic review and meta-analysis.

Although oxidative stress plays a major role in psoriasis, the association between oxidative stress biomarker levels and psoriasis in humans remains controversial. Relevant articles were retrieved by searching the following databases: PubMed, Web of Science, and EMBASE, without any time limit (updated March 10th, 2019). The pooled weighted mean difference (WMD) and 95% confidence interval (CI) for the total oxidant status (TOS), total antioxidant status (TAS), malondialdehyde (MDA), and catalase (CAT) were calculated for each study. Heterogeneity test, publication bias analysis, and sensitivity analysis were performed. A total of 28 case-control studies, containing a combined total of 2724 subjects (1485 psoriasis patients and 1239 healthy controls), were enrolled in this meta-analysis. The combined results showed a significant difference in the TAS levels (WMD = - 0.213 mmol/L, p < 0.001, 95% CI = - 0.216 to - 0.165), TOS levels (WMD = 2.196 µmol/L, p < 0.001, 95% CI = 1.667 to 2.726), MDA levels (WMD = 1.854 nmol/L, p < 0.001, 95% CI = 1.494 to 2.215), and CAT levels (WMD = - 22.341 kU/L, p = 0.008, 95% CI = - 38.934 to - 5.748) between psoriasis patients and controls. Moreover, the combined results showed a significant difference in the TAS, MDA, and CAT levels in patients with mild vs. moderate psoriasis and moderate vs. severe psoriasis. TAS and CAT levels in psoriasis patients were significantly lower than in healthy controls, whereas the TOS and MDA levels were significantly higher. Furthermore, the TAS, MDA, and CAT levels are associated with the severity of disease. These results indicate that redox imbalances play a major role in the pathogenesis of psoriasis.

Migration and transformation of nitrogen in bioretention system during rainfall runoff.

Bioretention systems have been extensively studied as a highly efficient technical measure to tackle the global threat of nitrogen pollution during global rainfall runoff. However, the migration and transformation of various forms nitrogen in bioretention system is unclear. So, in this paper, the bioretention systems with different flow regimes and planted configurations were designed to study the nitrogen removal performance and migration and transformation mechanism. The dynamic changes of NH4+-N and NO3--N were continuously monitored within 60 h after rainfall, and the abundance of 15N isotopes in soil layer NH4+-N was simultaneously measured. The results indicated that NH4+-N was mainly intercepted in soil layer in four constructed bioretention systems with similar removal efficiencies (95.42-97.69%). However, NO3--N was retained in submerged layer with significant different removal efficiencies (43.03-83.00%). After fitting calculation, the nitrification rate of NH4+-N (0.0626 mg kg-1 h-1) in soil was 5.31 times higher than that of the accumulation rate of NO3--N (0.0118 mg kg-1 h-1). During the elimination process of residual NH4+-N in soil, 41.46% removed by denitrification and plant absorption assimilation, another 57.28% stored in the form of organic nitrogen or inorganic nitrogen, only 1.26% leaked out. Based on this, the content variation of TN, NH4+-N and NO3--N could be analyzed by a system-wide and established the nitrogen balance model, which provides a new insight into the enhancement of nitrogen removal in the bioretention system.

Who eats what? Unravelling microbial conversion of coal to methane.

Microbial communities in subsurface coal seams are responsible for the conversion of coal organic matter to methane. This process has important implications for both energy production and our understanding of global carbon cycling. Despite the environmental and economic importance of this process, little is known about which components of the heterogeneous coal organic matter are biodegradable under methanogenic conditions. Similarly, little is known about which taxa in coal seams carry out the initial stages of coal organics degradation. To identify the biodegradable components of coal and the microorganisms responsible for their breakdown, a subbituminous coal was fractionated into a number of chemical compound classes which were used as the sole carbon source for growth by a coal seam microbial community. This study identifies 65 microbial taxa able to proliferate on specific coal fractions and demonstrates a surprising level of substrate specificity among members of this coal-degrading microbial consortia. Additionally, coal kerogen, the solvent-insoluble organic component of coal often considered recalcitrant to microbial degradation, appeared to be readily converted to methane by microbial degradation. These findings challenge our understanding of coal organic matter catabolism and provide insights into the catabolic roles of individual coal seam bacteria.

Circular RNA circ-DONSON facilitates gastric cancer growth and invasion via NURF complex dependent activation of transcription factor SOX4.

BACKGROUND: Circular RNAs (circRNAs) are a novel type of noncoding RNAs and play important roles in tumorigenesis, including gastric cancer (GC). However, the functions of most circRNAs remain poorly understood. In our study, we aimed to investigate the functions of a new circRNA circ-DONSON in GC progression. METHODS: The expression of circ-DONSON in gastric cancer tissues and adjacent normal tissues was analyzed by bioinformatics method, qRT-PCR, Northern blotting and in situ hybridization (ISH). The effects of circ-DONSON on GC cell proliferation, apoptosis, migration and invasion were measured by using CCK8, colony formation, EdU, immunofluorescence (IF), FACS and Transwell assays. qRT-PCR and Western blotting were utilized to validate how circ-DONSON regulates SOX4 expression. ChIP, DNA fluorescence in situ hybridization (DNA-FISH) and DNA accessibility assays were used to investigate how circ-DONSON regulates SOX4 transcription. The interaction between circ-DONSON and NURF complex was evaluated by mass spectrum, RNA immunoprecipitation (RIP), pulldown and EMSA assays. Xenograft mouse model was used to analyze the effect of circ-DONSON on GC growth in vivo. RESULTS: Elevated expression of circ-DONSON was observed in GC tissues and positively associated with advanced TNM stage and unfavorable prognosis. Silencing of circ-DONSON significantly suppressed the proliferation, migration and invasion of GC cells while promoting apoptosis. circ-DONSON was localized in the nucleus, recruited the NURF complex to SOX4 promoter and initiated its transcription. Silencing of the NURF complex subunit SNF2L, BPTF or RBBP4 similarly attenuated GC cell growth and increased apoptosis. circ-DONSON knockdown inhibited GC growth in vivo. CONCLUSION: circ-DONSON promotes GC progression through recruiting the NURF complex to initiate SOX4 expression.

Schwann cells protect against CaMKII- and PKA-dependent Acrylamide-induced Synapsin I phosphorylation.

OBJECTIVES: To explore the effects of Acrylamide (ACR), as well as the influence of Schwann cells (SCs), on the signal transduction pathway and phosphorylation of Synapsin I in a Human neuroblastoma cell line (NB-1). METHODS: NB-1s, NB-1s co-cultured with SCs, and a negative control group (NB-1 cells without ACR) were exposed to gradient concentrations of ACR for 48 h. Cell proliferation and viability were determined by MTT. Protein and mRNA expression levels of typical kinases (i.e., cAMP-dependent protein kinase [PKA], calcium/calmodulin-dependent protein kinase II [CaMKII], and mitogen-activated protein kinase-extracellular signal-regulated kinases [MAPK-Erk]), their phosphorylation status, as well as Synapsin I and its phosphorylation status, were tested by western blotting and polymerase chain reaction, respectively. Further, the effect of SCs on ACR-induced NB-1 cell toxicity was evaluated. RESULTS: (1) The MTT assay showed a sustained, dose- and time-dependent inhibition of NB-1s exposed to ACR. (2) ACR exposure increased the phosphorylation of CaMKII and PKA, which subsequently increased the phosphorylation of Synapsin I (at Serine603 [a substrate site of CaMKII] and Serine9 [a substrate site of PKA]). Pretreatment with CaMKII and PKA inhibitors blocked the ACR-mediated increase in phosphorylation. The above-described results were all significantly different when compared to the control group (p < 0.05). (3) When co-cultured with SCs, ACR-induced NB-1 inhibition was obviously decreased, and the trend of change of phosphorylated CaMKII, PKA, and Synapsin I were changed (first slightly increased and then decreased), which was inconsistent with what we observed in NB-1s cultured alone. CONCLUSIONS: The toxic effects of ACR on neurons may be mediated by CaMKII and PKA-dependent signaling pathways in which Synapsin I may act as a downstream effector. Furthermore, glial cells (SCs) may be able to prevent a certain degree of ACR-induced neuronal damage.

Analysis of TTSuV1b antibody in porcine serum and its correlation with four antibodies against common viral infectious diseases.

BACKGROUND: The purpose of the present study was to evaluate the correlation between Torque teno sus virus 1b (TTSuV1b) infection and other viral infections or vaccine immunization in conventional pigs. METHODS: With overexpressed and purified viral protein TTSuV1b as antigen, an indirect enzyme-linked immunosorbent assay (ELISA) method for detecting TTSuV1b antibody was established, which demonstrated great specificity and reproducibility. Porcine serum samples (n = 212) were tested using ELISA. Meanwhile, the antibodies against Classical Swine Fever Virus (CSFV), Pseudorabies Virus (PRV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and Porcine Circovirus 2 (PCV2) were also examined using the commercial kits. RESULTS: Statistical analysis indicated that the level of anti-TTSuV1b antibody was positively correlated with the level of anti-PCV2 antibody in a lesser extent; the level of antibodies against TTSuV1b or PCV2 were significantly lower in porcine serum with low level of TTSuV1b virus, implicating the potential consistency and synchronization in the mechanism of TTSuV1b and PCV2 infection. Whereas, antibodies against PRRSV or CSFV showed no statistical significance on comparison with anti-TTSuV1b antibody, implicating that in conventional pigs, the antibody level for PRRSV and CSFV were not significantly influenced by TTSuV1b infection. CONCLUSION: In conclusion, examination of anti-TTSuV1b antibody in porcine serum with the presently established ELISA method would serve as a supplementary approach for etiological investigation, and the combined statistical analysis of the antibodies against four other viruses might help to further understand the TTSuV1b infection as well as its pathogenicity.

[Research on olaquindox induced endoplasmic reticulum stress related apoptosis on nephrotoxicity].

OBJECTIVE: Renal tubular epithelial cell were exposed to olaquindox and detected the ROS and apoptosis related proteins, to investigate the renal tubular epithelial cell apoptosis through endoplasmic reticulum stress mediated pathway induced by olaquindox. METHODS: MTT assay (1, 2, 3, 4, 5, 6, 7and 8 µmol/ml olaquindox exposure) was used to detect the effects of olaquindox on renal tubular epithelial cell proliferation to determine test concentrations. Hoechst-33258 was used to detect morphological changes on apoptotic cells in each group. Flow cytometry method was applied to detect the apoptosis rate and intracellular reactive oxygen, and western blot assay was performed to detect the levels of endoplasmic reticulum stress-related apoptosis proteins, GRP78, GRP94 and CHOP. RESULTS: According to results of the MTT test, 1, 2, 3 and 4 µmol/ml olaquindox concentrations were determined for apoptosis analysis. With the increase of olaquindox concentration, apoptosis rate and levels of endoplasmic reticulum stress related apoptosis pathway protein GRP78, GRP94 and CHOP increased, levels of ROS were increased in every groups (P < 0.05) in 2 µmol/ml olaquindox groups and above. With the prolongation of olaquindox exposure, apoptosis rate and levels of endoplasmic reticulum stress related apoptosis pathway protein GRP78 and GRP94 increased in 12 and 24 h olaquindox exposure groups, whereas in groups of olaquindox exposed for 6, 12 and 24 h, levels of ROS and endoplasmic reticulum stress related apoptosis pathway protein CHOP increased (P < 0.05). CONCLUSION: Olaquindox can induce renal tubular epithelial cells to apoptosis and cause the renal toxicity, and the endoplasmic reticulum stress-related apoptosis maybe the associated toxicity pathway.

[Serological tests of the truncated TTSuV2 ORF1 recombinant proteins with the porcine sera].

OBJECTIVE: We studied the serological response between the special regions on the Torque teno sus virus 2 (TTSuV2) ORF1 coded protein and the porcine sera from conventional pigs. METHODS: Based on a Chinese TTSuV2 strain from Guangdong province, two overlapped virus proteins were expressed from Escherichia coli. Then, purified recombinant TTSuV2 ORF1a and TTSuV2 ORF1ab proteins were used as the antigens in the Western Blotting and ELISA assay. RESULTS: The recombinant TTSuV2 ORF1a and TTSuV2 ORF1ab proteins were identified with the special tag monoclonal antibody. The results of the ELISA tests shown that there were significant relationships between two groups of dates from the recombinant TTSuV2 ORF1a and TTSuV2 ORF1ab proteins antigenic assay. The results of the following Western Blotting assay indicated that the TTSuV2-specific IgG antibodies were contained in pig sera. CONCLUSION: The truncated TTSuV2 ORF1a protein (positions 168 to 346 corresponding to TTSuV2 GDIMA1) contains important B cell epitopes which can stimulate immune system antibody secretion. The truncated TTSuV2 ORF1a protein could be effective in TTSuV2 immunodiagnosis.