NRF1-mediated mitochondrial biogenesis antagonizes innate antiviral immunity.

Mitochondrial biogenesis is the process of generating new mitochondria to maintain cellular homeostasis. Here, we report that viruses exploit mitochondrial biogenesis to antagonize innate antiviral immunity. We found that nuclear respiratory factor-1 (NRF1), a vital transcriptional factor involved in nuclear-mitochondrial interactions, is essential for RNA (VSV) or DNA (HSV-1) virus-induced mitochondrial biogenesis. NRF1 deficiency resulted in enhanced innate immunity, a diminished viral load, and morbidity in mice. Mechanistically, the inhibition of NRF1-mediated mitochondrial biogenesis aggravated virus-induced mitochondrial damage, promoted the release of mitochondrial DNA (mtDNA), increased the production of mitochondrial reactive oxygen species (mtROS), and activated the innate immune response. Notably, virus-activated kinase TBK1 phosphorylated NRF1 at Ser318 and thereby triggered the inactivation of the NRF1-TFAM axis during HSV-1 infection. A knock-in (KI) strategy that mimicked TBK1-NRF1 signaling revealed that interrupting the TBK1-NRF1 connection ablated mtDNA release and thereby attenuated the HSV-1-induced innate antiviral response. Our study reveals a previously unidentified antiviral mechanism that utilizes a NRF1-mediated negative feedback loop to modulate mitochondrial biogenesis and antagonize innate immune response.

DIST: spatial transcriptomics enhancement using deep learning.

Spatially resolved transcriptomics technologies enable comprehensive measurement of gene expression patterns in the context of intact tissues. However, existing technologies suffer from either low resolution or shallow sequencing depth. Here, we present DIST, a deep learning-based method that imputes the gene expression profiles on unmeasured locations and enhances the gene expression for both original measured spots and imputed spots by self-supervised learning and transfer learning. We evaluate the performance of DIST for imputation, clustering, differential expression analysis and functional enrichment analysis. The results show that DIST can impute the gene expression accurately, enhance the gene expression for low-quality data, help detect more biological meaningful differentially expressed genes and pathways, therefore allow for deeper insights into the biological processes.

Randomized Double-Blind Study of the Effect of Injectate Temperature on Intrathecal Bupivacaine Dose Requirement in Spinal Anesthesia for Cesarean Delivery.

BACKGROUND: Increasing the temperature of intrathecal local anesthetics has been shown to increase the speed of onset and block height of spinal anesthesia. However, how this influences dose requirement has not been fully quantified. The aim of this study was to determine and compare the effective dose for anesthesia for cesarean delivery in 50% of patients (ED50) of intrathecal bupivacaine given at temperatures of 37 °C (body temperature) or 24 °C (room temperature). METHODS: Eighty healthy parturients having elective cesarean delivery under combined spinal-epidural anesthesia were randomly assigned to receive intrathecal hyperbaric bupivacaine stored at 37 °C (body temperature group) or 24 °C (room temperature group). The first subject in each group received a bupivacaine dose of 10 mg. The dose for each subsequent subject in each group was varied with an increment or decrement of 1 mg based on the response (effective or noneffective) of the previous subject. Patients for whom the dose was noneffective received epidural supplementation after data collection with lidocaine 2% as required until anesthesia was sufficient for surgery. Values for ED50 were calculated using modified up-down sequential analysis with probit analysis applied as a backup sensitivity analysis. These values were compared and the relative mean potency was calculated. RESULTS: The ED50 (mean [95% confidence interval, CI]) of intrathecal hyperbaric bupivacaine was lower in the body temperature group (6.7 [5.7-7.6] mg) compared with the room temperature group (8.1 [7.7-8.6] mg) (P < .05). The relative potency ratio for intrathecal bupivacaine for the room temperature group versus the body temperature group was 0.84 (95% CI, 0.77-0.93). CONCLUSIONS: Warming hyperbaric bupivacaine to body temperature reduced the dose requirement for spinal anesthesia for cesarean delivery by approximately 16% (95% CI, 7%-23%).

Visual psychosocial profiling of Chinese temporomandibular disorder pain patients and correlations with somatosensory function.

BACKGROUND: Psychosocial function of Chinese temporomandibular disorders (TMD) pain patients and the correlation with somatosensory function has not been sufficiently studied. OBJECTIVE: The study aims at assessing the psychosocial function of Chinese TMD pain patients by visualisation method and evaluating the correlations with somatosensory function quantitatively. METHODS: The Symptom Checklist 90 (SCL-90) questionnaire and standardised quantitative sensory testing (QST) were administered to 70 Chinese TMD pain patients and age- and gender-matched healthy controls (HCs). Of these, 40 TMD arthralgia patients received QST before and after medication. Psychosocial and somatosensory parameters were transformed into standardised scores. Differences within groups were assessed through t tests. Correlations between psychosocial and somatosensory profiles were explored through correlation analyses with Bonferroni correction for multiple comparisons. RESULTS: 100% of the Chinese TMD pain patients exhibited psychosocial distress in contrast to HCs. Anger and hostility showed negative correlation with the thermal nonnociceptive parameter (thermal sensory limen, p =.002) and nociceptive parameters (cold pain threshold and pain pressure threshold, p<.001). Correlation analysis indicated that cold detection threshold was negatively correlated with somatization and mechanical pain sensitivity had a negative correlation with anger and hostility through medical treatment (p <.001). CONCLUSIONS: Visual psychosocial profiles provided an easy overview of psychosocial function in Chinese TMD pain patients. Anger and hostility was associated with increased thermal nonnociceptive and nociceptive sensitivity to stimuli. Psychosocial distress might be negatively associated with TMD treatment response which indicated a possible need for psychological intervention during treatment.

Association of night-time sleep and daytime napping with painful temporomandibular disorder.

BACKGROUND: Painful temporomandibular disorder (TMD) is the common cause of chronic oro-facial pain, which may interfere with sleep. Previous studies have documented an association between sleep and TMD. OBJECTIVES: This study aimed to further explore the association of night-time sleep and daytime napping with painful TMD. METHODS: A total of 419 patients (aged 31.88 ± 11.54 years with women forming 85.4%) from a TMD/Orofacial Pain center were enrolled. Patients' sleep conditions were evaluated with the Pittsburgh Sleep Quality Index (PSQI) questionnaire, and information on night-time sleep duration, napping duration and napping frequency was interviewed. TMD was diagnosed according to the Diagnostic Criteria for TMD protocol and stratified into myalgia (muscle pain), arthralgia (joint pain) and combined (muscle and joint pain) subgroups. The severity of TMD was measured with the Fonseca Anamnestic Index (FAI) questionnaire. Restricted cubic spline (RCS) regression models were established to explore relationships between sleep and painful TMD subgroups. RESULTS: Patients with poor sleep quality (PSQI≥6) had higher FAI scores (median 60, p < .001) and higher proportions of painful TMDs. The myalgia subgroup had higher PSQI scores (median 8, p < .001) than the arthralgia subgroup. The RCS models indicated a non-linear relationship between night-time sleep duration and myalgia (p < .001), which was not observed in arthralgia. However, there were no significant findings concerning napping and painful TMD subgroups. CONCLUSION: This study found that the association between sleep and TMD is mainly related to painful TMD conditions, which are associated with night-time sleep duration.

Highly Sensitive and Selective Detection of Inorganic Phosphates in the Water Environment by Biosensors Based on Bioluminescence Resonance Energy Transfer.

The accurate detection of phosphate in water is very important to prevent water eutrophication and ensure the health of water quality. However, traditional phosphomolybdenum blue spectrophotometry is not sensitive, is time-consuming, and demands large amounts of chemical reagents. Therefore, highly sensitive, rapid, and environmentally friendly Pi detection methods are urgently needed. Here, we developed a bioluminescence resonance energy transfer (BRET)-based biosensor, which can detect Pi in water quickly, highly sensitively, and highly selectively. The NanoLuc and the Venus fluorescent protein were selected as the bioluminescence donor and energy acceptor, respectively. The best-performing BRET sensor variant, VenusΔC10-PΔC12-ΔN4Nluc, was identified by Pi-specific binding protein (PiBP) screening and systematic truncation. Single-factor experiments optimized the key parameters affecting the detection performance of the sensor. Under the optimal detection conditions, the detection limit of this method was 1.3 µg·L-1, the detection range was 3.3-434 µg·L-1, and it had excellent selectivity, repeatability, and stability. This low-cost and environment-friendly BRET sensor showed a good application prospect in real water quality detection.

Diagnostic accuracy of fused CBCT images in the evaluation of temporomandibular joint condylar bone resorption.

OBJECTIVES: To evaluate the diagnostic accuracy of fused CBCT images for patients with condylar bone resorption of temporomandibular joint (TMJ) osteoarthrosis. MATERIALS AND METHODS: Forty-two TMJs from twenty-one patients were included. Bone resorption of condyles evaluated by three experts was used as the reference standard. Three oral and maxillofacial radiology residents evaluated the resorption of condyles with a five-point scale for the four sets of images (two consecutive CBCT images without fusion, fused 2D cross-sectional images, fused 3D images, and combining fused 2D cross-sectional images and fused 3D images) randomly and independently. Each set of images was evaluated at least 1 week apart, and a second evaluation was performed 4 weeks later. Intraclass correlation coefficients were calculated to assess the intra- and inter-observer agreement. The areas under the ROC curves (AUCs) were compared among the four image sets using the Z test. RESULTS: Twenty-four TMJs were determined as condylar bone resorption, and eighteen were determined as no obvious change. The average AUC values from the three observers for the three fused image sets (0.94, 0.93, 0.93) were significantly higher than the image set without fusion (p < 0.01). The intra- and inter-observer agreement on the three fused image sets (0.70-0.89, 0.91-0.92) was higher than the image set without fusion (0.37-0.63, 0.75). CONCLUSIONS: Fused CBCT images of TMJ osteoarthrosis patients can intuitively display the condylar bone resorption and significantly improve the diagnostic accuracy. CLINICAL RELEVANCE: Fused CBCT images can help clinicians intuitively observe bone changes of the condyle in TMJ osteoarthrosis patients.

Antibacterial and anti-biofilm activities of probiotic Lactobacillus curvatus BSF206 and Pediococcus pentosaceus AC1-2 against Streptococcus mutans.

Streptococcus mutans (S. mutans) is the most important oral pathogenic bacterial cause of dental caries. Here we investigated the abilities of probiotic lactobacillus strains of Lactobacillus curvatus (L. curvatus) BSF206 and Pediococcus pentosaceus (P. pentosaceus) AC1-2 to control S. mutans. Both probiotic strains are acid and bile salt tolerant and are resistant to hydrogen peroxide and lysozyme to promote their survival within the oral environment. In addition, both strains are highly hydrophobic and are also capable of engaging in electrostatic interactions. These properties enhance abilities of both strains to adhere to gingival epithelial cells and HT-29 for improved colonization of oral tissues, while also enabling these probiotics auto-aggregate and to form aggregates with S. mutans that both may prevent S. mutans from colonizing oral tissues and facilitate the clearance of the cariogenic bacteria from the mouth during swallowing of food and saliva. Furthermore, results presented herein revealed that L. curvatus BSF206 and P. pentosaceus AC1-2 effectively inhibited S. mutans activities (biofilm formation, secretion of extracellular matrix components, synthesis of water-insoluble glucans) and led to downregulation of expression of key S. mutans genes involved in biofilm production (gtfA, gtfB, ftf, brpA). Taken together, these results indicate that L. curvatus BSF206 and P. pentosaceus AC1-2 can inhibit S. mutans biofilm formation as a new strategy for preventing dental caries.

Analysis of MR Signs to Distinguish Between ARCO Stages 2 and 3A in Osteonecrosis of the Femoral Head.

BACKGROUND: MRI is the most effective diagnostic tool of osteonecrosis of the femoral head (ONFH), especially for early diagnosis, but its detection of subchondral or cortical fractures is less accurate than CT. Therefore, it is difficult to accurately stage ONFH in the peri-collapse period by MRI. PURPOSE: To improve the accuracy of MR for distinguishing between Association Research Circulation Osseous (ARCO) stages 2 and 3A in ONFH. STUDY TYPE: Retrospective. SUBJECTS: One hundred and fifty five cases of ARCO stage 2/3A of ONFH underwent MR examinations, M/F = 72/83. CT was used as reference standard for collapse, which was decided by an orthopedist and a radiologist in consultation. FIELD STRENGTH/SEQUENCE: 3 T/axial and coronal T1 -weighted Turbo Spin Echo (T1 W TSE) sequence, axial T2 -weighted fat-saturated (T2 W FS) TSE sequence, and coronal proton density-weighted imaging (PDWI)-FS-Dixon fat/water image. ASSESSMENT: Five potential MR signs (the maximum width of the necrotic-viable interface, bone marrow edema (BME), irregular articular surface of the femoral head, T2 heterogeneous high signal, and the absence of a necrotic-viable interface with the morphology of closed loop) were evaluated blindly by five radiologists independently and the total scores of different combinations of MR signs were calculated. STATISTICAL TESTS: Mann-Whitney U test and Chi-square test were used to evaluate age, gender, and MR signs differences between the two groups. ROC curve was used to access the distinguishing value of MR signs. The consistency of the five radiologists was analyzed by intraclass correlation coefficient. RESULTS: The area under the curve of the combined MR signs 2 for distinguishing between ARCO stages 2 and 3A was the greatest (0.967), sensitivity and specificity were 100.00% and 88.71% respectively, and greater than 1 was the threshold. DATA CONCLUSION: Combined MR signs 2 has great values in distinguishing between ARCO stages 2 and 3A in ONFH, thus helping clinical therapy. LEVEL OF EVIDENCE: 4 TECHNICAL EFFICACY: Stage 2.

Diagnostic value of serum sphingolipids in patients with colorectal cancer.

Background: Abnormal lipid metabolism affects the regulation of tumor progression, though use of serum lipids and sphingolipids for disease progression identification is uncertain. Methods: Serum samples from 51 healthy volunteers and 76 patients were collected and analyzed by liquid chromatography tandem mass spectrometry. Results: Levels of serum total cholesterol and high-density lipoprotein were significantly lower in colorectal cancer patients. Multivariate analysis demonstrated distinct sphingolipid profiles between healthy individuals and patients. Of 106 sphingolipids, 15 metabolites that showed statistical significance were selected, and receiver operating characteristic analysis of these metabolites yielded an area under the curve of 0.868 to 0.9 by machine learning algorithms for distinguishing colorectal cancer from a healthy status. Conclusions: Healthy individuals, polyps patients and colorectal cancer patients have different serum sphingolipid signatures. Serum sphingolipids might be used as biomarkers for early detection or prediction of colorectal cancer.

Association Between Thromboelastography and Coronary Heart Disease.

BACKGROUND Thromboelastography (TEG) is a novel blood viscoelasticity detection method revealing blood coagulation status and has been reported to be helpful in predicting clinical outcomes in patients with cardiovascular diseases (CVD). In this study, we aimed to investigate the association between TEG and CVD. MATERIAL AND METHODS A single-center case-control study was performed. Individuals who took TEG tests at Tongji Hospital in Wuhan, China from 2015 to 2019 were included. The nearest-neighbor Mahalanobis matching with replacement, within propensity score calipers of 0.25 was used to control the covariate imbalance between CVD patients and controls. Logistic regression analyses were conducted to assess the relationship between TEG and CVD. Subgroup and sensitivity analyses were performed to evaluate the robustness of the association between TEG and CVD. RESULTS After matching, a total of 151 participants were included in this study, with 83 patients having CVD (49 patients having coronary heart disease [CHD] and 34 patients having an ischemic stroke). By comparison, CHD patients had a significantly higher maximum amplitude (MA) (P=0.02) than controls. After multivariable adjustment, MA (OR 1.11, 95% CI 1.01-1.24, P=0.04) was independently associated with CHD. The association between MA and CHD remained robust across subgroups and in sensitivity analyses. CONCLUSIONS The current study suggests that MA is significantly associated with CHD. Enhanced platelet reactivity as described by high MA might be associated with risk of CHD. The exact role of MA in the measurement of CHD risk needs to be further examined in large-scale prospective cohort studies.

Characterizing dynamic changes of plasma cell-free Echinococcus granulosus DNA before and after cystic echinococcosis treatment initiation.

Over one million people are living with cystic echinococcosis (CE) and alveolar echinococcosis (AE). For CE, long-term albendazole treatment is often needed, which requires regular follow-up. Follow-up is mainly through imaging which is insensitive to subtle changes and subjective to experience. We investigated the changes of Echinococcus granulosus (Eg) cell-free DNA (cfDNA) in plasma of CE patients before and after albendazole treatment to evaluate its potential as an objective marker for treatment follow-up. Plasma samples of nine CE patients were collected before and after treatment. We identified Eg cfDNA from every sample through high-throughput sequencing. Eg cfDNA concentration and fragment length increased significantly after the treatment period. Ultrasound examination before and after the treatment initiation reflected the drug effects to a certain extent, as the cyst size of four patients reduced. Our findings indicated that Eg cfDNA from plasma could be a potential marker in the monitoring of CE treatment.

Genetic characteristics of human papillomavirus type 16, 18, 52 and 58 in southern China.

Persistent infections of high-risk human papillomaviruses (HPVs) are the leading cause of cervical cancers. We collected cervical exfoliated cell samples from females in Changsha city, Hunan Province and obtained 338 viral genomes of four major HPV types, including HPV 16 (n = 82), 18 (n = 35), 52 (n = 121) and 58 (n = 100). The lineage/sublineage distribution of the four HPVs confirmed previous epidemiological reports, with the predominant prevailing sublineage as A4 (50%), A1 (37%) and A3 (13%) for HPV16, A1 (83%) for HPV18, B2 (86%) for HPV52 and A1 (65%), A3 (19%) and A2 (12%) for HPV58. We also identified two potentially novel HPV18 sublineages, i.e. A6 and A7. Virus mutation analysis further revealed the presence of HPV16 and HPV58 sublineages associated with potentially high oncogenicity. These findings expanded our knowledge of the HPV genetic diversity in China, providing valuable evidence to facilitate HPV DNA screening, vaccine effectiveness evaluation and control strategy development.

The variants at FLNA and FLNB contribute to the susceptibility of hypertension and stroke with differentially expressed mRNA.

BACKGROUND: Filamin A and filamin B were involved in vascular development and remodeling. Herein, it is important to explore the associations of FLNA and FLNB variants with hypertension and stroke. METHODS: The associations of two single-nucleotide polymorphisms (SNPs) at FLNA and five SNPs at FLNB with hypertension and stroke were examined in two case-control studies and a cohort study in Chinese Han population. Risks were estimated as odds ratio (OR) and hazard ratio (HR) by Logistic and Cox regression analysis respectively. In addition, filamin B, FLNA and FLNB mRNA expression were measured. RESULTS: In the case-control study of hypertension, FLNA rs2070816 (CT + TT vs. CC) and rs2070829 (CG + GG vs. CC) were significantly associated with hypertension in <55 years group (OR = 1.338, P = 0.018; OR = 1.615, P = 0.005) and FLNB rs839240 (AG + GG vs. AA) was significantly associated with hypertension in females (OR = 0.828, P = 0.041) and nonsmokers (OR = 0.829, P = 0.020). In the follow-up study, rs2070829 GG genotype carriers presented a higher risk of hypertension than CC/CG in males (HR = 1.737, P = 0.014) and smokers (HR = 1.949, P = 0.012). In the case-control study of stroke, FLNB rs1131356 variation was significantly associated with ischemic stroke (IS) and intracerebral hemorrhage (ICH), ORs of additive model were 1.342 and 1.451, with P values of 0.001 and 0.007. The FLNA transcript 2, FLNB transcript 3, transcript 4 mRNA, and filamin B expression levels were significantly different between IS cases and hypertension controls and among the genotypes of rs839240 in hypertensive individuals (P < 0.05). CONCLUSIONS: Our findings support the genetic contribution of FLNA and FLNB to hypertension, and stroke with differentially mRNA expression.

Calorie restriction conferred improvement effect on long-term rehabilitation of ischemic stroke via gut microbiota.

Calorie restriction can modulate the gut microbiota and protect against many diseases including ischemic stroke. However, the role of calorie-restriction-induced microbiota alteration remained unknown in ischemic stroke rehabilitation. Here we conducted 30% reduction of caloric intake on mice for four weeks, to evaluate its role on ischemic stroke rehabilitation. Significantly, this calorie restriction led to better long-term rehabilitation in comparison of normal control. Notably, the transplantation of gut microbiome from calorie-restriction-treated mice to post-stroke mice was eligible to obtain better long-term rehabilitation of stroke mice. Bifidobacterium identified by 16 S ribosomal RNA sequencing were enriched in those of calorie-restriction mice. Then we administrated Bifidobacterium to stroke mice and found Bifidobacterium treatment could successfully improve the long-term rehabilitation of cerebral ischemia mice. Furthermore, the metabolomics analysis revealed a panel of upshifting metabolites, suggesting that calorie restriction greatly altered the gut microbiota composition and its metabolism. Hence, we discovered the novel effect of CR on long-term rehabilitation of ischemic stroke and the underlying role of gut microbiota, which might provide novel thoughts for the clinical post-stroke rehabilitation.

Roxarsone transformation and its impacts on soil enzyme activity in paddy soils: A new insight into water flooding effects.

The aromatic arsenical roxarsone (ROX) has been used as feed additive for decades worldwide. The past or present application of animal manure containing ROX in paddy fields results in arsenic (As) accumulation in rice grain. However, the degradation and transformation mechanisms of ROX in paddy soil which determine As bioavailability and uptake by rice are still unclear. The current study investigated the variation of As speciation and soil enzyme activities in ROX-treated soils under flooded and non-flooded conditions for six months. Our results showed that 70.2% of ROX persisted in non-flooded paddy soils after 180 d while ROX degraded completely within 7 d in flooded soils. The rapid degradation of ROX under flooded conditions owed to the enhanced biotic transformation that was caused by the low Eh and the predominant presence of Clostridium spp. and Bacillus spp. ROX was not only transformed to As(III) and As(V) in non-flooded soils but also to 3-amino-4-hydroxyphenylarsonic acid and methyl arsenicals in flooded soils. The degradation products significantly inhibited soil enzyme activities for 7-30 d, but the inhibition effects disappeared after 90 d due to the sorption of transformed As products to amorphous Fe oxides. This study provides new insights into the flooding effect on ROX fate in paddy fields, which is important for the management of animal waste and risk control on polluted sites.

4D-QSAR Molecular Modeling and Analysis of Flavonoid Derivatives as Acetylcholinesterase Inhibitors.

Flavonoids are potential strikingly natural compounds with antioxidant activity and acetylcholinesterase (AChE) inhibitory activity for treating Alzheimer's disease (AD). In present study, in line with our interests in flavonoid derivatives as AChE inhibitors, a four-dimensional quantitative structure-activity relationship (4D-QSAR) molecular model was proposed. The data required to perform 4D-QSAR analysis includes 52 compounds reported in the literature, usually analogs, and their measured biological activities in a common assay. The model was generated by a complete set of 4D-QSAR program which was written by our group. The best model was found after trying multiple experiments. It had a good predictive ability with the cross-validation correlation coefficient Q2 = 0.77, the internal validation correlation coefficient R2 = 0.954, and the external validation correlation coefficient R2pred = 0.715. The molecular docking analysis was also carried out to understand exceedingly the interactions between flavonoids and the AChE targets, which was in good agreement with the 4D-QSAR model. Based on the information provided by the 4D-QSAR model and molecular docking analysis, the idea for optimizing the structures of flavonoids as AChE inhibitors was put forward which maybe provide theoretical guidance for the research and development of new AChE inhibitors.

HTRA1 Variants and the Interaction with Smoking Confer the Genetic Susceptibility to Ischemic Stroke.

High temperature requirement protein A1 (HtrA1) was identified as the causative gene of autosomal recessive arteriopathy and associated with lacunar ischemic stroke (IS) in European. This study aimed at evaluating the association of HTRA1 with IS and four tagging single-nucleotide polymorphisms (SNPs) were genotyped in a cohort of 4,098 Chinese. The mRNA level of HTRA1 in 72 IS cases and 72 hypertension controls were measured and compared. In whole population, SNP rs2268350 (C>T) was significantly associated with IS incidence (P=0.034). Stratification analysis observed significant association of rs2268350 in male, smoking and drinking populations, rs2672587 (C>G) in smoking and nonsmoking populations and rs3793917 (C>G) in smoking, nonsmoking and nondrinking populations with stroke respectively (P<0.05). The additive interaction and multiplicative interaction between rs2268350 and smoking were both of significant (P<0.05) after adjustment for the covariates. There was a cumulated risk of IS among genotypes of rs3793917 (P=0.009) and rs2672587 (P=0.047) in smoking population. The mRNA level of HTRA1 in non-smokers with rs2268350 CC was significantly higher than smokers with rs2268350 CT/TT (P=0.046) in IS cases. Our findings support that HTRA1 confers the genetic susceptibility to IS and smoking might modify the genetic effect of HTRA1 on IS by suppressing HTRA1 mRNA expression.

Molecular Cloning and Characterization of FAD6 Gene from Chia (Salvia hispanica L.).

Plastidial Δ12 fatty acid desaturase (FAD6) is a key enzyme for linoleic acid (LA) and α-linolenic acid (ALA) biosynthesis. Chia (Salvia hispanica L.) is a revived omega-3 plant source that is richest in ALA level. In this study, based on the RACE method, one full-length cDNA sequence encoding FAD6, named ShFAD6, was isolated from chia. There exist three alternative transcription start sites and five alternative poly(A) tailing sites in ShFAD6. The 5'UTR of ShFAD6 contains a purine-stretch of 44 bp. ShFAD6 has an ORF of 1335 bp encoding a 444 aa protein of 51.33 kDa. ShFAD6 contains a conserved Delta12-FADS-like domain together with three strong trans-membrane helices and three histidine motifs. There also exists a chloroplast transmit peptide in ShFAD6 N-terminal. Phylogenetic analyses validated its identity of dicot FAD6 protein and suggested some critical evolutionary features of plant FAD6 genes. Heterologous yeast expression confirmed the catalytic activity of ShFAD6. The qRT-PCR assay showed that ShFAD6 is mainly expressed in leaves, stems, flowers, buds and early-stage seeds, and also responded to various stresses and hormone treatments. Under Sclerotinia infection, qRT-PCR and fluorescence imaging illustrated the possible correlation of ShFAD6 expression and photosynthesis. This study provides insight for further function study of ShFAD6 in oil quality improvement in staple oilseed crops as well as stress response and adaptation in plants.

Dynamic sulfur-iron cycle promoted phosphorus mobilization in sediments driven by the algae decomposition.

Direct evidence of the algae bloom in eutrophic freshwater lakes on sulfur cycle and the subsequent iron oxide reduction and the iron oxides-bound phosphate (Fe-P) release in sediments is lacking. In this study, microcosms experiment was carried out to investigate the dynamic variations of S, Fe and P species in the water column and sediments as well as the sulfate reducing bacteria (SRB) abundance variation in the sediments during algae decomposition. The sulfate reduction was stimulated by the algae decomposition, which resulted in dramatic sulfate decline, sulfide increase and SRB growth. In addition, large amounts of acid volatile sulfide (AVS), pyrite sulfur (Pyrite-S) and elemental sulfur (S0) accumulated in the sediment. In particular, the contents of sedimentary Fe(II) and Pyrite-S in surface sediments continuously accumulated until the end of the experiment. Moreover, the terminal Fe-P content reduced by 35.4% compared with the initial concentration at high algae density group. These results suggested the irreversible reduction of iron oxides and revealed iron chemical reduction mediated by sulfide during algae decomposition. In addition, the connection of sulfur-iron cycle and the significant promotion of Fe-P mobilization in sediments was established, which should be paid more attention in the eutrophic freshwater ecosystems.