Spatial and Single-Cell Transcriptomics Reveals the Regional Division of the Spatial Structure of MASH Fibrosis.

OBJECTIVE: To elucidate the regional distribution of metabolic dysfunction-associated steatohepatitis (MASH) fibrosis within the liver and to identify potential therapeutic targets for MASH fibrosis. METHODS: Liver sections from healthy controls, patients with simple steatosis and MASH patients were analysed using spatial transcriptomics integrated with single-cell RNA-seq. RESULTS: Spatial transcriptomics analysis of liver tissues revealed that the fibrotic region (Cluster 9) was primarily distributed in lobules, with some fibrosis also found in the surrounding area. Integration of the single-cell-sequencing data set (GSE189175) showed a greater proportion of inflammatory cells (Kupffer cells and T cells) and myofibroblasts in MASH. Six genes, showing high- or low-specific expression in Cluster 9, namely, ADAMTSL2, PTGDS, S100A6, PPP1R1A, ASS1 and G6PC, were identified in combination with pathology. The average expression levels of ADAMTSL2, PTGDS and S100A6 on the pathological HE staining map were positively correlated with the increase in the degree of fibrosis and aligned strongly with the distribution of fibrosis. ADAMTSL2+ myofibroblasts play a role in TNF signalling pathways and in the production of ECM structural components. Pseudotime analysis indicated that in the early stages of MASH, infiltration by T cells and Kupffer cells triggers a significant inflammatory response. Subsequently, this inflammation leads to the activation of hepatic stellate cells (HSCs), transforming them into myofibroblasts and promoting the development of liver fibrosis. CONCLUSION: This study is the first to characterise lineage-specific changes in gene expression, subpopulation composition, and pseudotime analysis in MASH fibrosis and reveals potential therapeutic targets for this condition.

Application of DynaCT angiographic reconstruction in balloon pulmonary angioplasty.

OBJECTIVES: To investigate the feasibility and accuracy of balloon pulmonary angioplasty (BPA) using DynaCT angiographic reconstruction guidance. METHODS: Thirty-four BPAs (23 CTEPH patients) targeting 175 pulmonary arteries were included. Eleven BPAs (2D group) were guided by DSA two-dimensional angiography. Another twenty-three BPAs (3D group) were guided using DynaCT angiographic reconstruction. The volume rendering (VR) method was used to obtain a three-dimensional image of the blood vessels. This image was used as a reference to continue BPA treatment under the guidance of vascular three-dimensional reconstruction technology. Procedure durations and radiation exposure data were compared between the two groups using Mann-Whitney U test. RESULTS: Using the DynaCT angiographic reconstruction technique, more target vessels were treated in a single BPA procedure (5.83 ± 2.33 vs 3.73 ± 1.10 vessels per BPA, p = 0.008) in a shorter operation time (3.58 ± 0.61 vs 4.49 ± 0.91 h, p = 0.002). Overall, the dose area product (DAP) was significantly higher for the 2D group than for the 3D group (13,901.82 ± 5549.69 vs 4682.82 ± 1950.64, p < 0.001). The use of the DynaCT angiographic reconstruction technique to guide BPA required a lower dose of contrast agent (225.22 ± 48.70 vs 292.73 ± 76.82 mL, p = 0.013) and less radiation exposure. CONCLUSIONS: The use of DynaCT angiographic reconstruction guidance in patients undergoing BPA is feasible and accurate. Images of DynaCT angiographic reconstruction may be beneficial for optimizing the operative process in BPA with reduced radiation exposure. KEY POINTS: • BPA guidance by DynaCT angiographic reconstruction is feasible and accurate. • DynaCT angiographic reconstruction may be beneficial for optimizing the operative process. • DynaCT angiographic reconstruction can reduce patient radiation dose due to multi-times of BPA sessions.

Mutation screening of Chinese Treacher Collins syndrome patients identified novel TCOF1 mutations.

Treacher Collins syndrome (TCS) (OMIM 154500) is a rare congenital craniofacial disorder with an autosomal dominant manner of inheritance in most cases. To date, three pathogenic genes (TCOF1, POLR1D and POLR1C) have been identified. In this study, we conducted mutational analysis on Chinese TCS patients to reveal a mutational spectrum of known causative genes and show phenotype-genotype data to provide more information for gene counselling and future studies on the pathogenesis of TCS. Twenty-two TCS patients were recruited from two tertiary referral centres, and Sanger sequencing for the coding exons and exon-intron boundaries of TCOF1, POLR1D and POLR1C was performed. For patients without small variants, further copy number variations (CNVs) analysis was conducted using high-density SNP array platforms. The Sanger sequencing overall mutation detection rate was as high as 86.3% (19/22) for our cohort. Fifteen TCOF1 pathogenic variants, including ten novel mutations, were identified in nineteen patients. No causative mutations in POLR1D and POLR1C genes and no CNVs mutations were detected. A suspected autosomal dominant inheritance case that implies germinal mosaicism was described. Our study confirmed that TCOF1 was the main disease-causing gene for the Chinese TCS population and revealed its mutation spectrum. We also addressed the need for more studies of mosaicism in TCS cases, which could explain the mechanism of autosomal dominant inheritance in TCS cases and benefit the prevention of TCS.

Clinical Features of Non-Alcoholic Fatty Liver Disease in the Non-Lean Population.

INTRODUCTION: The prevalence of non-alcoholic fatty liver disease (NAFLD) in non-lean patients is significantly increased, and obesity significantly increases the risk of cirrhosis and HCC in NAFLD patients. However, whether there is a difference in clinical manifestations of NAFLD between overweight and obesity remains unclear. The objective of this study was to assess the clinical and histological features of NAFLD among a non-lean population. METHODS: Current study enrolled consecutive non-lean (body mass index [BMI] >23 kg/m2) patients with NAFLD and available liver biopsy results. Patients were stratified by BMI into two groups for the comparison of their clinical and histological variables, which included the overweight (BMI 23∼<28 kg/m2) and the obese (BMI ≥28 kg/m2). Risk factors for moderate to severe fibrosis (stage >1) were also analyzed through the logistic regression model. RESULTS: Among 184 non-lean patients with metabolic-associated fatty liver disease enrolled, 65 and 119 were overweight and obese, respectively. Patients in the obesity group had a significantly lower level of gamma-glutamyl transpeptidase, higher levels of platelet, glucose, prothrombin time, and more common of moderate to severe inflammatory activity when compared to those in the overweight group. However, a significant low frequency of moderate to severe fibrosis was found in the obesity group versus the overweight group (19.33% vs. 40.00%, p = 0.002). Binary logistics regression analysis of fibrosis found that aspartate transaminase (AST), BMI, alanine transaminase (ALT), and cholesterol (CHOL) were independent predictors for moderate to severe fibrosis in non-lean patients with NAFLD. Compared with the traditional fibrosis-4 (AUC = 0.77) and aminotransferase to platelet ratio index (AUC = 0.79) indexes, the combined index based on AST, BMI, ALT, and CHOL was more accurate in predicting moderate to severe fibrosis in non-lean patients with NAFLD (AUC = 0.87). CONCLUSIONS: Clinical and histological features differed between obesity and overweight patients with NAFLD. When compared to the traditional serum markers, the combination index including AST, BMI, ALT, and CHOL provided a better model to predict moderate to severe fibrosis in non-lean patients with NAFLD.

[The correlation between brain natriuretic peptide and invasive hemodynamic parameters and their value in prognosis of patients with noncardiac shock].

OBJECTIVE: To examine the correlation between the plasma level of brain natriuretic peptide (BNP) and the hemodynamic parameters collected through Swan-Ganz flowing balloon catheter procedure in patients with noncardiac shock, in order to evaluate the potential for BNP to be used as prognostic indicator. METHODS: The plasma BNP and invasive hemodynamic parameters data [central venous pressure (CVP), pulmonary arterial pressure (PAP), pulmonary capillary wedge pressure (PCWP), and cardiac output (CO)] were collected from 21 noncardiac shock patients received Swan-Ganz catheterization throughout a continuous surveillance for 3 days. The BNP, CVP, PAP, PCWP, CO in survivors (n = 8) and non-survivors (n = 13) were compared and the correlation between the value of BNP and the invasive hemodynamic parameters were analyzed using multiple regression. RESULTS: The mean value of BNP (ng/L) was significantly higher in non-survivors (708.06 ± 242.58 vs. 317.05 ± 140.21, P < 0.05). In day 1, no significant difference was found in any hemodynamic parameters between non-survivors and survivors. But in day 3, the non-survivors were found to have significantly higher CVP (mm Hg, 1 mm Hg = 0.133 kPa: 13.64 ± 4.00 vs. 9.92 ± 1.26, P < 0.05) and lower CO (L/min: 4.61 ± 2.06 vs. 6.95 ± 1.28, P < 0.05). The differences in PAP (mm Hg: 20.84 ± 8.48 vs. 16.82 ± 4.97) and PCWP (mm Hg: 13.60 ± 5.71 vs. 12.72 ± 4.98) remained insignificant (both P > 0.05) between the two groups. The correlation between BNP and the invasive hemodynamic parameters was modest there was no correlation between BNP and CVP, PAP, PCWP, CO (r = 0.157, 0.306, 0.229, -0.269, P = 0.16, 0.25, 0.09, 0.12). CONCLUSIONS: In patients with shock, both plasma BNP and invasive hemodynamic examination showed certain prognostic value. But in noncardiac shock cases, the increased BNP did not correlate with heart function, therefore it could not replace the Swan-Ganz catheter data to guide the treatment in these patients.

[Acupoint selection rules of post-stroke cognitive impairment treated with acupuncture based on ancient and modern medical record cloud platform].

OBJECTIVE: To analyze the acupoint selection rules of post-stroke cognitive impairment (PSCI) treated with acupuncture by ancient and modern medical record cloud platform (V1.5). METHODS: The published randomized controlled trial (RCT) literature of PSCI treated with acupuncture was retrieved from CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase, Cochrane Library and Web of Science, and the data was extracted to establish prescription database. The ancient and modern medical record cloud platform (V1.5) was used to analyze the data. RESULTS: A total of 185 articles was included, involving 91 acupoints, the total using frequency was 1174 times. The top 5 acupoints in frequency of PSCI treated with acupuncture were Baihui (GV 20, 151 times), Shenting (GV 24, 97 times), Sishencong (EX-HN 1, 83 times), Neiguan (PC 6, 69 times) and Sanyinjiao (SP 6, 64 times); the most involved meridian was the governor vessel, and the generally used acupoints were mainly distributed in the head, face and neck. The top 5 acupoint combinations in frequency were Baihui (GV 20)-Shenting (GV 24, 89 times), Baihui (GV 20)-Sishencong (EX-HN 1, 79 times), Baihui (GV 20)-Neiguan (PC 6, 59 times), Baihui (GV 20)-Sanyinjiao (SP 6, 56 times) and Baihui (GV 20)-Zusanli (ST 36, 51 times). The acupoint combination with the strongest association was Shenting (GV 24)→Baihui (GV 20). There were 6 acupoint cluster groups according to the cluster analysis, and the main core prescription was Baihui (GV 20), Shenting (GV 24), Sishencong (EX-HN 1), Zusanli (ST 36), Neiguan (PC 6) and Sanyinjiao (SP 6). CONCLUSION: Acupoints on the governor vessel, and distributed in head, face and neck are the main acupoints for PSCI treated with acupuncture, Baihui (GV 20), Shenting (GV 24), Sishencong (EX-HN 1), Zusanli (ST 36), Neiguan (PC 6) and Sanyinjiao (SP 6) can be used as the main acupoints, but it is still necessary to combine with syndrome differentiation.

SfgA Renders Aspergillus flavus More Stable to the External Environment.

sfgA is known as a key negative transcriptional regulator gene of asexual sporulation and sterigmatocystin production in Aspergillus nidulans. However, here, we found that the homolog sfgA gene shows a broad and complex regulatory role in governing growth, conidiation, sclerotia formation, secondary metabolism, and environmental stress responses in Aspergillus flavus. When sfgA was deleted in A. flavus, the fungal growth was slowed, but the conidiation was significantly increased, and the sclerotia formation displayed different behavior at different temperatures, which increased at 30 °C but decreased at 36 °C. In addition, sfgA regulated aflatoxin biosynthesis in a complex way that was associated with the changes in cultured conditions, and the increased production of aflatoxin in the ∆sfgA mutant was associated with a decrease in sclerotia size. Furthermore, the ∆sfgA mutant exhibited sensitivity to osmotic, oxidative, and cell wall stresses but still produced dense conidia. Transcriptome data indicated that numerous development- and secondary-metabolism-related genes were expressed differently when sfgA was deleted. Additionally, we also found that sfgA functions downstream of fluG in A. flavus, which is consistent with the genetic position in FluG-mediated conidiation in A. nidulans. Collectively, sfgA plays a critical role in the development, secondary metabolism, and stress responses of A. flavus, and sfgA renders A. flavus more stable to the external environment.

Accounting and red uction path of carbon emission firom facility agriculture in China.

Facility agriculture, a typical agricultural production management mode, could affect carbon cycle due to its unique production environmental conditions and highly intensive utilization. With the five main sources as accounting objects, including agricultural film investment, energy consumption, pesticide and fertilizer application, CO2 fertilizer application, and facility soil, we estimated the amount and intensity of carbon emission of three facility agriculture (continuous greenhouse, solar greenhouse, and plastic greenhouse) in 31 provinces in 2018. The results showed that the total carbon emission of facility agriculture in China was 210.3817 million t CO2e, with the three facility agriculture types of plastic greenhouse, solar greenhouse, and continuous greenhouse accounting for 60.2%, 37.4% and 2.4%, respectively. Carbon emission of facility agriculture was mainly contributed by soil greenhouse gas, agricultural film and supplies investment. Carbon emission per unit area of continuous greenhouses was significantly lower than that of solar and plastic greenhouses. The scientific capital allocation rate and facility agriculture scale were the two main factors influencing the carbon emission in facility agriculture. Based on all the results, we presented the carbon emission reduction path from the three perspectives of improving the scientific investment, material consumption utilization rate, and facility area utilization rate of facility agriculture.

Effects of different acupuncture treatment methods on post-stroke cognitive impairment: study protocol for a multicenter randomized controlled trial.

BACKGROUND: Cognitive impairment is a common dysfunction after stroke that seriously affects the overall recovery of patients. Cognitive rehabilitation training is currently the main treatment to improve cognitive function, but its curative effect is limited. Acupuncture is a core component of traditional Chinese medicine (TCM), and some previous clinical studies have shown that it might be effective in treating post-stroke cognitive impairment (PSCI), but further evidence from large-sample studies is needed. The overall objective of this trial is to obtain further data to develop an optimized acupuncture treatment for PSCI by comparing the effects of different acupuncture treatment methods on cognitive function in PSCI patients. METHODS/DESIGN: In this multicenter, prospective, randomized controlled trial, 206 eligible stroke inpatients who meet the trial criteria will be randomly assigned to 2 groups: an electroacupuncture (EA) plus needle retention (NR) group and an EA group. Both groups of patients will undergo the same routine cognitive rehabilitation treatments. All treatments will be given 5 times per week for 8 weeks. The primary outcomes will be assessed using the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment scale (MOCA). The secondary outcome will be measured by the Barthel Index (BI). All outcomes will be evaluated at baseline, week 4, week 8, and the third and sixth month after the end of treatment. DISCUSSION: Our aim is to evaluate the effects of two different acupuncture treatment methods for treating PSCI patients. This study is expected to provide data to be used in developing an optimized acupuncture treatment method for PSCI treatment. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR1900027849. Registered on 30 November 2019, http://www.chictr.org.cn/showproj.aspx?proj=46316.

Electroacupuncture improves learning and memory functions in a rat cerebral ischemia/reperfusion injury model through PI3K/Akt signaling pathway activation.

Electroacupuncture has been widely used to treat cognitive impairment after cerebral ischemia, but the underlying mechanism has not yet been fully elucidated. Studies have shown that autophagy plays an important role in the formation and development of cognitive impairment, and the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway plays an important role in autophagy regulation. To investigate the role played by the PI3K/Akt signaling pathway in the electroacupuncture treatment of cerebral ischemia/reperfusion rat models, we first established a rat model of cerebral ischemia/reperfusion through the occlusion of the middle cerebral artery using the suture method. Starting at 2 hours after modeling, electroacupuncture was delivered at the Shenting (GV24) and Baihui (GV20) acupoints, with a dilatational wave (1-20 Hz frequency, 2 mA intensity, 6 V peak voltage), for 30 minutes/day over 8 consecutive days. Our results showed that electroacupuncture reduced the infarct volume in a rat model of cerebral ischemia/reperfusion injury, increased the mRNA expression levels of the PI3K/Akt signaling pathway-related factors Beclin-1, mammalian target of rapamycin (mTOR), and PI3K, increased the protein expression levels of phosphorylated Akt, Beclin-1, PI3K, and mTOR in the ischemic cerebral cortex, and simultaneously reduced p53 mRNA and protein expression levels. In the Morris water maze test, the latency to find the hidden platform was significantly shortened among rats subjected to electroacupuncture stimulation compared with rats without electroacupuncture stimulation. In the spatial probe test, the number of times that a rat crossed the target quadrant was increased in rats subjected to electroacupuncture stimulation compared with rats without electroacupuncture stimulation. Electroacupuncture stimulation applied to the Shenting (GV24) and Baihui (GV20) acupoints activated the PI3K/Akt signaling pathway and improved rat learning and memory impairment. This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Henan University of Traditional Chinese Medicine, China (approval No. 8150150901) on March 10, 2016.

The comparative effectiveness of traditional Chinese medicine exercise therapies in elderly people with mild cognitive impairment: A protocol for a network meta-analysis.

BACKGROUND: Mild cognitive impairment (MCI) in the elderly is a health problem worldwide. Several clinical trials indicated that traditional Chinese medicine (TCM) exercise therapies can effectively improve MCI, such as Tai Ji, Baduan jin exercise, Liuzi jue, and finger exercise. However, there is still controversy over which therapy is the best for elderly MCI patients. In this study, we aimed to systematically evaluate and compare the effectiveness and safety of these 4 TCM exercise therapies in elderly patients with MCI. METHODS: The Web of Science, PubMed, EMBASE, Cochrane Central Register of Controlled Trials, Clinical Trials, China National Knowledge Infrastructure, Wangfang database, and Chinese Biomedical Medicine will be comprehensively searched to collect all randomized controlled trials which included elderly participants with MCI receiving TCM exercise therapies through July 2020. Two reviewers will independently screen and evaluate each included study and extract the outcome indexes. ADDIS 1.16.8 software will be used for the network meta-analysis and STATA 14 software will be used for drawing network evidence plots and funnel plots. RESULTS: We will use the Bayesian statistical model to conduct a network meta-analysis to rank the effectiveness and safety of these 4 interventions, and use the GRADE approach to interpret the results. CONCLUSION: This network meta-analysis will find out the optimal treatment plan for MCI and provide evidence-based bias for clinical treatments decision-making. PROTOCOL REGISTRATION NUMBER: INPLASY202070006.

The Kinetochore Protein Spc105, a Novel Interaction Partner of LaeA, Regulates Development and Secondary Metabolism in Aspergillus flavus.

Nuclear protein LaeA is known as the global regulator of secondary metabolism in Aspergillus. LaeA connects with VeA and VelB to form a heterotrimeric complex, which coordinates fungal development and secondary metabolism. Here, we describe a new interaction partner of LaeA, the kinetochore protein Spc105, from the aflatoxin-producing fungus Aspergillus flavus. We showed that in addition to involvement in nuclear division, Spc105 is required for normal conidiophore development and sclerotia production of A. flavus. Moreover, Spc105 positively regulates the production of secondary metabolites such as aflatoxin and kojic acid, and negatively regulates the production of cyclopiazonic acid. Transcriptome analysis of the Δspc105 strain revealed that 23 backbone genes were differentially expressed, corresponding to 19 of the predicted 56 secondary metabolite gene clusters, suggesting a broad regulatory role of Spc105 in secondary metabolism. Notably, the reduced expression of laeA in our transcriptome data led to the discovery of the correlation between Spc105 and LaeA, and double mutant analysis indicated a functional interdependence between Spc105 and LaeA. Further, in vitro and in vivo protein interaction assays revealed that Spc105 interacts directly with the S-adenosylmethionine (SAM)-binding domain of LaeA, and that the leucine zipper motif in Spc105 is required for this interaction. The Spc105-LaeA interaction identified in our study indicates a cooperative interplay of distinct regulators in A. flavus, providing new insights into fungal secondary metabolism regulation networks.

Clinical study of optical coherence tomography in the diagnosis of peripheral pulmonary artery thrombus.

BACKGROUND: Peripheral pulmonary artery thrombus (PPT) is common in the clinic. However, due to the lack of an ideal diagnostic tool, PPT cannot be quickly diagnosed and effectively treated at present. Optical coherence tomography (OCT) is a new intravascular imaging technique that is characterized by high image resolution. This technique is suitable for small vessel imaging and has the ability to distinguish red and white thrombi. OBJECTIVE: This study aimed to evaluate the value of OCT in the diagnosis of PPT and in identifying the nature of thrombi by comparing the difference in sensitivity between OCT and selective pulmonary angiography (SPA). METHODS: Highly suspected PPT patients were enrolled in this study. Pulmonary ventilation/perfusion (V/Q) mismatch pulmonary segments or peripheral pulmonary arteries were chosen. SPA was performed first, followed by OCT imaging. The diagnostic standard of thrombus with SPA was an intraluminal filling defect. The procedure and criteria for OCT diagnosis of thrombus were previously introduced in intracoronary OCT thrombus images. The diameter of the imaging vessels was measured, and they were grouped according to diameter. The diameter was <2mm in the distal segment group, 2-3mm in the middle segment group, and >3mm in the proximal segment group. The recognition abilities of intravascular thrombus with the different diameters of these two techniques were compared. Patients with obvious clinical symptoms and more red thromboses revealed by OCT were given standardized anticoagulant therapy for half a year. The clinical symptoms, 6-minute walking test and changes in the thrombus in the OCT images of these patients before and after treatment were observed. RESULTS: A total of 22 patients with highly suspected PPT were suggested to undergo V/Q inspection. Finally, 12 patients were eligible for the study. SPA and OCT were performed in 61 peripheral pulmonary arteries in all 12 patients. The ideal SPA and OCT images obtained from a total of 76 blood vessel segments were suitable for comparative analysis. A total of 62 thrombi were found by SPA. Among these, eight thrombi were in the distal segment, 29 thrombi were in the middle segment, and 25 thrombi were in the proximal segment. A total of 81 thrombi were found by OCT, among which 22 thrombi were in the distal segment, 31 were in the middle segment, and 28 were in the proximal segment. There was a significant difference between two groups in the distal segment group (P=0.013), while there was no significant difference between two groups in the middle segment group or the proximal segment group (P>0.05). In addition to all the thrombi found by SPA, OCT found other thrombi that were missed by SPA. According to previous OCT images for determining the nature of thrombi, OCT revealed 81 thrombi, of which 48 (59%) were red thrombi and 33 (41%) were white thrombi. Then, seven patients who had obvious clinical symptoms and more red thrombi in the peripheral pulmonary artery were given anticoagulant therapy for six months. After treatment, these symptoms were improved, oxygenation indexes increased, and the six-minute walking test was extended in all these patients. After anticoagulation therapy, five patients underwent OCT review. These OCT images were matched and compared before and after anticoagulation therapy. The results revealed that most of the thrombi had disappeared, and a small amount of red thrombi turned white as the volume reduced. The mean lumen area before and after treatment was 2.05±1.03mm2 and 2.86±1.24mm2, respectively, and the difference was statistically significant (P=0.035). CONCLUSION: OCT can clearly show the structure of the lumen and the wall of the peripheral pulmonary artery tube. The sensitivity of the diagnosis of PPT with a diameter of <2mm was higher than that of SPA. Moreover, OCT has the ability to distinguish between red and white thrombi, which is of guiding significance in anticoagulant therapy.

The Antioxidant Gallic Acid Inhibits Aflatoxin Formation in Aspergillus flavus by Modulating Transcription Factors FarB and CreA.

Aflatoxin biosynthesis is correlated with oxidative stress and is proposed to function as a secondary defense mechanism to redundant intracellular reactive oxygen species (ROS). We find that the antioxidant gallic acid inhibits aflatoxin formation and growth in Aspergillus flavus in a dose-dependent manner. Global expression analysis (RNA-Seq) of gallic acid-treated A. flavus showed that 0.8% (w/v) gallic acid revealed two possible routes of aflatoxin inhibition. Gallic acid significantly inhibited the expression of farB, encoding a transcription factor that participates in peroxisomal fatty acid β-oxidation, a fundamental contributor to aflatoxin production. Secondly, the carbon repression regulator encoding gene, creA, was significantly down regulated by gallic acid treatment. CreA is necessary for aflatoxin synthesis, and aflatoxin biosynthesis genes were significantly downregulated in ∆creA mutants. In addition, the results of antioxidant enzyme activities and the lipid oxidation levels coupled with RNA-Seq data of antioxidant genes indicated that gallic acid may reduce oxidative stress through the glutathione- and thioredoxin-dependent systems in A. flavus.

A cytosine methyltransferase ortholog dmtA is involved in the sensitivity of Aspergillus flavus to environmental stresses.

DNA methylation is an important epigenetic modification that depends on DNA methyltransferases (DMT). However, the filamentous fungus Aspergillus flavus has no detectable methylation, and role of a DMT homologue, DmtA, is undefined. Here we describe the role of the dmtA gene responding to changes in the environment by comparing knockout, point mutation, over-expression and wild type strains. Deletion of dmtA differentially affected conidia development in a media-dependent fashion, which suggests that dmtA plays an important role in conidiation. Furthermore, ΔdmtA strains lost the capacity to form the resistant structure, sclerotia, and alleviated sensitivity to several stress conditions, such as high osmotic pressure, hypoxia, low water activity and a high calcium concentration. We also noticed that deletion of dmtA and mutation C377S in DmtA negatively affected aflatoxin production and down regulated the expression of some early (fas-1, pksA, nor-1), middle and late (nor-A, ver-1, avnA, omtB) genes in the aflatoxin biosynthetic cluster. Finally, we found that all tested strains showed a similar phenotype when treated with 5-azacytidine. Our results indicate that the dmtA gene is important in regulation of aflatoxin biosynthesis and for A. flavus to adapt to stressful environments and for survival, although it may hold no apparent function in DNA methylation.

Minocycline attenuates cognitive impairment and restrains oxidative stress in the hippocampus of rats with chronic cerebral hypoperfusion.

OBJECTIVE: Nitric oxide (NO) was speculated to play an important role in the pathophysiology of cerebral ischemia. Minocycline, a tetracycline derivative, reduced inflammation and protected against cerebral ischemia. To study the neuroprotection mechanism of minocycline for vascular dementia, the influences of minocycline on expressions of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) were observed in the brains of Wistar rats. METHODS: The vascular dementia rat model was established by permanent bilateral common carotid arteries occlusion (BCCAO). Wistar rats were divideded into 3 groups randomly: sham-operation group (S group), vascular dementia model group (M group), and minocycline treatment group (MT group). The behaviour was tested with Morris water maze and open-field task. Expressions of iNOS and eNOS were measured by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR). The optical density value was measured by imaging analysis. Percentage of positive cells with iNOS and eNOS expression was analyzed with optical microscope. RESULTS: Minocycline attenuated cognitive impairment. Inducible NOS was significantly down-regulated in MT group, compared with that in M group (P < 0.01), while eNOS was significantly up-regulated, compared with that in M group (P < 0.01). The expressions of iNOS and eNOS in M and MT groups were higher than those in S group (P < 0.01). CONCLUSION: Minocycline can down-regulate the expression of iNOS and up-regulate the expression of eNOS in vascular dementia, which restrains apoptosis and oxidative stress to protect neural function.