The effects of Bacillus subtilis on the immunity, mucosal tissue morphology, immune-related gene transcriptions, and intestinal microbiota in flounder (Paralichthys olivaceus) with two feeding methods: Continuous versus discontinuous feeding.

Probiotics as dietary additives can improve weight gain, feed efficiency, and disease resistance in cultured fish. In this research, we evaluated and compared the effects of Bacillus subtilis on immunity, mucosal tissue morphology, immune-related gene transcriptions, and intestinal microbiota in flounder (Paralichthys olivaceus) by a 30-day feeding experiment based on a continuous feeding schedule (E1) and a discontinuous feeding schedule (E2). As a result, the use of B. subtilis exerted the best positive effects on survival rate, enzyme activity, mucosal tissue morphology, immune-related gene transcriptions, and intestinal microbiota in flounders. Alkaline phosphatase (AKP), lysozyme (LZM), and superoxide dismutase (SOD) activities in the liver of E2 were higher than those of E1 (P < 0.05). Furthermore, the villi length in the intestinal tract and the fold length in the stomach of E2 were also higher than in E1 (P < 0.05). The il-1 expression levels in the spleen were significantly increased in E2 (P < 0.05) compared to E1. We performed 16 S rRNA sequencing analysis to find that Bacillus in E1 (1.06%) and E2 (1.01%) had higher relative abundances than in E0 (0.053%) at the end of the experiments, indicating that short-term application of B. subtilis with the continuous or discontinuous feeding method can allow both the adaptation of the ecosystem to the presence of probiotics by the establishment of new species in the gut microbiota and the ability these new probiotic species to perform corresponding functions. No significant differences in the ability of probiotic establishment were observed between E1 and E2. Our findings provided a unique perspective to explore the mechanism of immune enhancement with probiotics and to screen the optimal administration strategy in aquaculture application for probiotic use. Together, these results point to some level of enhancement in immune status by continuous and discontinuous feeding after a short-term feeding period, which could be used as a prophylactic strategy for flounder health management.

Effects of the water-soluble fraction of diesel oil on the sera biochemical indicators, histological changes, and immune responses of black rockfish Sebastes schlegelii.

In order to understand the physiological and immune responses of Sebastes schlegelii to the water-soluble fraction of diesel oil (WSD), S. schlegelii were used as the experimental objects to study the effect of WSD on the sera biochemical indicators, histological changes, and immune responses. Significant differences in sera biochemical indicators were observed in S. schlegelii after WSD exposure. The alkaline phosphatase (ALP), glucose (GLU), and globulin (GLB) were reduced by 3.51-fold, 3.12- fold, and 1.58-fold, respectively; however, K+ was increased by 3.55-fold. The results of HE staining showed that interstitial congestion was observed in the liver; the secondary lamellae deformity and hyperplasia, epithelial lifting, the primary lamellae hyperplasia, and aneurism were observed in the gill. Epidermis thickness increased, and epidermal hyperplasia in the skin was shown. The length of the secondary lamellae shortened significantly after WSD exposure. The results of AB-PAS staining showed that three different types of mucous cells were observed in the gill, and a significant increase in the number of all three types of mucous cells was observed after WSD exposure (P < 0.05). In addition, the results of the relative mRNA expressions in the liver of eleven immune-related genes showed that the relative expression levels of IL-1ß, IL-8, TNF receptor, BAFF, C1s, C1r, and MyD88 in the WSD group were substantially higher than those in the LPS group (P < 0.05), and the relative expression of caspase 10 was significantly lower than that in the LPS group (P < 0.05). At the same time, no significant differences were observed in the relative expression levels of IL-1, TNFα, and C1inh between the two groups (P > 0.05). This study was expected to provide essential data for health assessments of S. schlegelii and establish the foundation for the immune-related researches of S. schlegelii after WSD exposure.

Comparative study of peripheral blood cells in two marine fishes (Synechogobius hasta and Sebastes schlegelii): Morphological and cytochemical characterization.

The morphology, cell size and relative number of peripheral blood cells in two teleosts, Synechogobius hasta and Sebastes schlegelii, were compared using different staining methods. The results showed significant differences in cell size. The percentage of monocytes in S. hasta was greater than that in S. schlegelii (P < 0.01); however, the opposite results were obtained in the percentage of lymphocytes and thrombocytes. The two fishes shared common cytochemical-staining results, which showed that both erythrocytes were negative for all cytochemical staining; monocytes were strongly positive for PAS and positive for SBB and NAE; lymphocytes were negative for SBB, POX and NAE; neutrophils were positive for SBB and NAE; and thrombocytes were negative for SBB, ALP, POX and NAE. However, species specificity existed in the cytochemical properties. For S. hasta, monocytes were strongly positive for ALP and ACP; lymphocytes were strongly positive for ALP and weakly positive for ACP; neutrophils were strongly positive for ACP and POX; and thrombocytes were weakly positive for PAS and positive for ACP. Unlike S. hasta, monocytes were strongly positive for ACP and positive for ALP in S. schlegelii; lymphocytes were positive for ALP and partially positive for PAS; neutrophils were positive for ACP and POX; and thrombocytes were negative for PAS and ACP. The POX activity of monocytes in S. hasta was greater than that in S. schlegelii (P < 0.05), while the POX activity of neutrophils and the NAE activity of monocytes in S. hasta were significantly greater than those in S. schlegelii (P < 0.01). The results of this study can be used as a reference for the construction of haematological parameters in S. hasta and S. schlegelii for the assessment of fish health and can provide a research basis for fish diseases and immunity.

Identification and characterization of MYC transcription factors in Taxus sp.

Myelocytomatosis (MYC) transcription factors (TFs) are key regulators of the jasmonic acid (JA) signaling pathway. In cell cultures, methyl jasmonate (MeJA) can improve the production of taxol, which is a complex terpenoid compound with an intense antitumor activity. However, the functions of MYC genes in Taxus sp. (yew trees) remain poorly known. Based on Taxus sp. transcriptome changes induced by MeJA, a TcMYC gene was isolated in a previous study. Here, we further characterized the TcMYC TF encoded by that gene and four other yew MYC TFs previously obtained. Three yew MYC TFs had the typical basic helix-loop-helix (bHLH)-MYC_N region, but the other two MYC did not, although all five presented the bHLH domain. TcMYC was localized to the nuclei, and phylogenetic analysis indicated that the yew MYC TFs were closely related to Arabidopsis thaliana MYC1/2 and maize R protein. The yeast one-hybrid assay showed that TcMYC binds the G-box of the promoter of taxane 5α-hydroxylase. Transcript levels of TcMYC revealed that TcMYC was highly expressed in xylem and leaves, and up-regulated by drought and high-salinity stresses. Coronatine (COR) has recently been used as a new elicitor to improve the production of taxol in cell cultures; TcMYC was strongly expressed at 2 and 4 h after COR treatment, but decreased at 12 and 24 h. Overall, the results obtained here provide new insights into the potential regulatory roles of MYC TFs on taxol biosynthesis in yew trees.

The value of hip circumference/heightx ratio for identifying childhood hypertension.

To investigate the value of hip circumference related indexes for identifying childhood hypertension. In 2011, 1,352 Han children aged 7-12 years were recruited in our study. Hypertension was defined as systolic blood pressure or diastolic blood pressure ≥95th percentile for all three screenings. We set the power value of the hip circumference/heightx ratio (x = 0, 0.8, 1 and 1.5) and studied the association with blood pressure. Hip circumference, hip circumference/height0.8, hip circumference/height and hip circumference/height1.5 all showed a positive correlation with systolic blood pressure and diastolic blood pressure(P < 0.05). Area under the curve (AUC) was used to evaluate the abilities of hip circumference related indexes. Hip circumference/height0.8, hip circumference/height and hip circumference/height1.5 were not superior to hip circumference. The present study demonstrates that hip circumference measurement is a helpful tool to detect the presence of hypertension in Han children 7-12 years old.

Unified Deep Learning Architecture for Modeling Biology Sequence.

Prediction of the spatial structure or function of biological macromolecules based on their sequences remains an important challenge in bioinformatics. When modeling biological sequences using traditional sequencing models, long-range interaction, complicated and variable output of labeled structures, and variable length of biological sequences usually lead to different solutions on a case-by-case basis. This study proposed a unified deep learning architecture based on long short-term memory or a gated recurrent unit to capture long-range interactions. The architecture designs the optional reshape operator to adapt to the diversity of the output labels and implements a training algorithm to support the training of sequence models capable of processing variable-length sequences. The merging and pooling operators enhances the ability of capturing short-range interactions between basic units of biological sequences. The proposed deep-learning architecture and its training algorithm might be capable of solving currently variable biological sequence-modeling problems under a unified framework. We validated the model on one of the most difficult biological sequence-modeling problems, protein residue interaction prediction. The results indicate that the accuracy of obtaining the residue interactions of the model exceeded popular approaches by 10 percent on multiple widely-used benchmarks.

Intravoxel incoherent motion diffusion-weighted imaging for discriminating the pathological response to neoadjuvant chemoradiotherapy in locally advanced rectal cancer.

To investigate the usefulness of intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) in discriminating the pathological complete response (pCR) to neoadjuvant chemoradiotherapy (nCRT) in locally advanced rectal cancer (LARC), 42 patients underwent preoperative IVIM-DWI before (pre-nCRT) and after nCRT (post-nCRT). The values of pre-nCRT and post-nCRT IVIM-DWI parameters (ADC, D, D* and f), together with the percentage changes (∆% parametric value) induced by nCRT, were compared between the pCR (tumour regression grade [TRG] 4) and non-pCR (TRG 0, 1, 2 or 3) groups and between the GR (TRG 3 or 4) and PR (TRG 0, 1 or 2) groups based on the Dworak TRG system. After nCRT, the ADC and D values for LARC increased significantly (all P < 0.05). The TRG score revealed a positive correlation with pref (r = 0.357, P = 0.020), postD (r = 0.551, P < 0.001) and Δ%D (r = 0.605, P < 0.001). The pCR group (n = 10) had higher preD*, pref, postD, ∆%ADC and ∆%D values than the non-pCR group (n = 32) (all P < 0.05). The GR group (n = 15) exhibited higher postD, ∆%ADC and ∆%D values than the PR group (n = 27) (all P < 0.05). Based on ROC analysis, ∆%D had a higher area under the curve value than ∆%ADC (P = 0.009) in discriminating the pCR from non-pCR groups. In conclusion, IVIM-DWI may be helpful in identifying the pCR to nCRT for LARC and is more accurate than traditional DWI.

Intravoxel incoherent motion MRI for predicting early response to induction chemotherapy and chemoradiotherapy in patients with nasopharyngeal carcinoma.

BACKGROUND: To investigate the value of intravoxel incoherent motion (IVIM) diffusion-weighted magnetic resonance imaging (DW-MRI) in predicting the early response to induction chemotherapy (IC) and chemoradiotherapy (CRT) in nasopharyngeal carcinoma (NPC). METHODS: Fifty NPC patients who received IC and CRT underwent an IVIM DW-MRI on a 1.5-Tesla MRI scanner. The pretreatment and posttreatment (20 days after IC initiation) IVIM-based parameters (ADC, D, D*, and f), and their percentage changes (△%), were compared between the effective (complete response or partial response) and ineffective (stable disease) groups based on RECIST 1.1, and between the residual and nonresidual groups. RESULTS: None of the perfusion-related parameter' values showed significant differences between the effective and ineffective groups (p values for pref, postf, △%f, preD*, postD*, and △%D* were 0.364, 0.129, 0.792, 0.804, 0.167, and 0.428, respectively), or between the residual and nonresidual groups (P values for pref, postf, △%f, preD*, postD*, and △%D* were 0.328, 0.776, 0.546, 0.558, 0.214, and 0.414, respectively). The ineffective group exhibited higher preADC, higher preD and lower △%D values than the effective group (all P < 0.001). The nonresidual group had lower preD, lower preADC and higher △%D values (all P < 0.05) than the residual group. △%D had the highest area under curve (0.859) in predicting the response to IC, whereas preD had the highest area under curve (0.841) in predicting tumor residue after CRT. CONCLUSION: Diffusion-related IVIM-based parameters might be more helpful than perfusion-related parameters in predicting the early effects of IC and CRT for NPC.

[Clinical analysis of relapsing anti-N-methyl-D-aspartate receptor encephalitis].

OBJECTIVE: With the discovery of more patients with anti-N-methyl-D-aspartate (anti- NMDAR) encephalitis, frequent clinical relapses pose a new challenge to neurologists. METHODS: Retrospective reviews were conducted for 16 hospitalized patients with relapsing anti-NMDAR encephalitis at our hospital from June 2011 until November 2014. Their clinical data including symptoms, cerebrospinal fluid (CSF) profiles, neuroimaging findings and relapsing treatment were compared with those initial episodes. RESULTS: There were 11 females and 5 males with a mean onset time of 21.2 (10-34) years. For initial episodes, the mean number of major symptoms was 5. 8 and the mean modified Rankin score (mRS) 4.56. And 7 (43.8%) cases were admitted into intensive care unit (ICU). All received first-line immunotherapy and only one case second-line immunotherapy. Ovarian teratoma was detected and resected in only one case of initial episode. Among 32 relapses, 8 cases (50% ) had multiple (2-4) relapses. There was a median delay of 5.0 (0.5-18) months for relapses. Relapses were common upon pausing or reducing immunotherapy, usually monotherapy with corticosteroids. Compared with initial episodes, relapses were less severe (mean mRS 2.69, mean number of symptom 2.59) and only 2 cases were admitted into ICU during relapses. Presentation of relapses were partial symptoms of initial episode. However, two patients had new symptoms of brain stem involvement. Brain magnetic resonance imaging (MRI) of 8 cases showed abnormality initially during initial episode and disappearance at relapses while new lesions appeared in 7 patients including 3 cases with CNS demyelinating features of central nervous system ( CNS) on MRI. The positivity rate of anti-NMDAR antibody was 100% in CSF and 53.1% in sera during relapses. Anti-AQP4 and NMO-Ig were positive in one case with brain stem involvement. All cases received first-line immunotherapy and 12 chronic second-line immunotherapy. Two cases of ovarian teratoma were detected on reassessment during relapse and then resected. CONCLUSION: Inadequacy of second-line and chronic immunotherapy, occult teratoma and potential demyelination may contribute to a relapse of anti-NMDAR encephalitis. And its proper management should follow the recommendations of guidelines.

Role of salivary anti-SSA/B antibodies for diagnosing primary Sjögren's syndrome.

UNLABELLED: The diagnosis of primary Sjögren's syndrome (pSS) is complex, and the saliva test is a potential method to improve the existing diagnostic criteria. OBJECTIVE: To estimate the diagnostic accuracy of salivary anti-SSA/B antibodies in primary Sjögren's syndrome (pSS), and to analyze their correlations with clinical and laboratory profiles. STUDY DESIGN: This study enrolled 100 pSS patients and 140 non-pSS controls, including 40 rheumatoid arthritis (RA) patients, 40 systemic lupus erythematosus (SLE) patients, and 60 healthy controls. Unstimulated whole saliva and stimulated parotid saliva samples were collected from the subjects. Salivary anti-SSA/B antibodies were measured using an enzyme-linked immunosorbent assay (ELISA). Clinical and laboratory data were retrieved from the medical records. RESULTS: In the pSS group, the sensitivity of anti-SSA and anti-SSB antibodies in whole saliva was 49% and 29%, respectively, and the specificity was 87.5% and 95%. The sensitivity of anti-SSA and anti-SSB antibodies in parotid saliva was 32% and 8%, respectively, and the specificity was 95.52% and 97.86%, respectively. In the pSS group, the diagnostic accuracy of anti-SSA/B antibodies in whole saliva was significantly higher than in parotid saliva (p<0.05), but was significantly lower than in serum (p<0.05). The salivary flow rate in the pSS group positive for whole salivary anti-SSA was significantly lower than in the negative group (p<0.05). The prevalence of rheumatoid factor and antinuclear factor were significantly higher in salivary SSB-positive pSS patients than in SSB-negative patients (p<0.05). CONCLUSIONS: Compared to parotid saliva, whole saliva is a more suitable diagnostic fluid. Using salivary anti-SSA/B antibodies as a single test item is insufficient given the relatively low sensitivity. Further studies should investigate the possibility of combining tests for different salivary autoantibodies as a method for diagnosing pSS.

Stimuli-responsive one-dimensional copolymer nanostructures fabricated by metallogel template polymerization and their adsorption of aspirin.

pH responsive poly(N,N'-methylenebisacrylamide-co-4-vinylpyridine) (P(MBA-4VP)) one dimensional (1D) nanostructures have been prepared by metallogel template copolymerization, which was carried out in an Ag(i)-coordinated organogel with benzoyl peroxide (BPO) as the initiator. The product has been characterized using infrared spectroscopy, scanning electron microscopy and transmission electron microscopy. The experimental results reveal that the gel fiber is a crucial template for polymerization. Due to the degradation of the template in copolymerization, nanofibers of metallogel were transcribed to copolymer nanowires. The introduction of co-monomer 4-vinylpyridine (4VP) imparts to the 1D copolymer nanostructures pH sensitivity and the possible use as an adsorption material of aspirin. Adsorbed 1D copolymer nanostructures could be regenerated using proton solvent, acid medium and salt solution. In addition, silver nanoparticle loaded copolymer nanowires have been produced from the reduction of silver ions instead of template removal, where silver ions act both as the template and as the nanoparticle growth substrate.

The expression of LytM is down-regulated by RNAIII in Staphylococcus aureus.

LytM is one kind of autolysin expressed in Staphylococcus aureus. It can hydrolyze peptide bond of pentaglycine, which is the component of cell wall. Until now, the regulation on the expression of LytM is still unclear. Here, we found that the level of LytM in the RNAIII mutant was increased. RNAIII is an important small regulatory RNA in S. aureus. Our further investigation revealed that RNAIII can interact with the 5'UTR of lytM mRNA and block the ribosome binding sites [RBS]. So our results identified that LytM was the new target of RNAIII.

[Clinical assessment of oral diagnostic items in 2002 classification criteria for primary Sjogren's syndrome].

OBJECTIVE: To evaluate the diagnostic value of each item in 2002 international classification criteria for primary Sjogren's syndrome (pSS) in clinically diagnosed cases. METHODS: All patients were from the Department of Oral Medicine of Peking University School of Stomatology from 2005 to 2010. Their clinical manifestations and lab testing results met the standard criteria and the diagnosis was established according to international classification criteria (2002). We retrospectively collected all of the clinical and information and did the data analysis. RESULTS: A total of 148 pSS patients were included in the study. When the oral evaluations were performed, 98.0% of the patients complained dry mouth, 96.6% of them had decreased non-stimulated salivary flow rate, and 60.1% had positive parotid sialography results, which was consistent with diagnosis. It was found out in the lab testing that 79.7% of the patients had positive anti-SSA antibody, and 75.0% of them had elevated globulin level. 20.3% of the patients were finally diagnosed by performing the lower lip biopsy. CONCLUSION: When applying 2002 international classification criteria for the diagnosis of pSS patients, oral symptoms and unstimulated salivary flow rate are feasible and sensitive, which play an important and indicative role in the diagnosis of pSS.

G1359A polymorphism in the cannabinoid receptor-1 gene is associated with the presence of coronary artery disease in patients with type 2 diabetes.

OBJECTIVE: Previous studies demonstrated that G1359A polymorphism of cannabinoid receptor-1 (CNR1) was associated with cardiovascular risk factors including obesity, insulin resistance, dyslipidemia, and inflammation, which are also risk factors for developing type 2 diabetes mellitus (T2DM). Therefore, this study was aimed to determine whether G1359A polymorphism of CNR1 is associated with T2DM and the presence of coronary artery disease (CAD) in patients with T2DM. METHODS: A total of 450 patients with T2DM (259 patients with CAD and 191 patients without CAD) and 94 healthy subjects were genotyped using polymerase chain reaction and restriction fragment length polymorphism method. RESULTS: No significant differences in genotype frequency of CNR1 were found between normal controls and patients with T2DM without CAD. GG genotype frequency of CNR1 was significantly higher in the patients with T2DM with CAD compared with those without CAD and healthy subjects (P = 0.003 and P = 0.005, respectively). Unconditional logistic regression analysis revealed that GG genotype was significantly associated with the presence of CAD in the patients with T2DM compared with GA and AA genotypes (odds ratio, 2.632; 95% confidence interval, 1.481-4.678; P < 0.001). In addition, GG genotype of CNR1 was significantly correlated with elevated levels of body mass index, systolic blood pressure, homeostasis model assessment of insulin resistance, and C-reactive protein, as well as decreased levels of high-density lipoprotein cholesterol in patients with T2DM. CONCLUSIONS: G1359A polymorphism of CNR1 may be not associated with T2DM but may contribute to the genetic risk for the presence of CAD in patients with T2DM of Chinese Han population.

The study of insulin resistance and soluble intercellular adhesion molecule-1 in normotensive adolescents with a family history of hypertension.

To investigate the changes of insulin resistance and soluble intercellular adhesion molecule-1 (sICAM-1) in normotensive adolescents with a family history of hypertension (FH) and to analyse the related factors. The study included 142 normotensive adolescents aged 13-15 years. On the basis of an FH, the samples were divided into positive FH (FH(+) group) and negative FH groups (FH(-) group). Anthropometric measurements included height, weight, waist circumference (WC) and blood pressure (BP). Blood samples were collected and fasting plasma glucose (FPG), serum lipids, true insulin (TI) and sICAM-1 were assayed. The FH(+) group had significantly higher levels of FPG, TI, homeostasis model assessment of insulin resistance (HOMA-IR) and sICAM-1 than the FH(-) group (P<0.01). HOMA-IR showed positive correlation with body mass index (BMI; r=0.405), WC (r=0.416), systolic BP (SBP; r=0.165), FPG (r=0.427), triglyceride (r=0.195), TI (r=0.993) and negative correlation with high density lipoprotein cholesterol (r=-0.196) (P<0.05). sICAM-1 showed a positive correlation with triglyceride (r=0.239, P<0.01). In multiple linear regression analysis, WC (beta=0.024, P=0.000) and FH (beta=0.467, P=0.000) were significant independent predictors for HOMA-IR, and triglyceride (beta=0.328, P=0.007) and FH (beta=0.395, P=0.016) were significant independent predictors for sICAM-1. These results indicate that insulin resistance and endothelial activation were found in normotensive adolescents with an FH. We should make great efforts to alleviate the insulin resistance and endothelial activation by diet and exercise, and it would reduce the incidence of hypertension in adolescents with an FH.

The prevalence of LRRK2 Gly2385Arg variant in Chinese Han population with Parkinson's disease.

We conducted a case-control study to determine the prevalence of the LRRK2 Gly2385Arg variant in patients with Parkinson's disease in Han population in mainland China. Heterozygous LRRK2 Gly2385Arg variant was identified in 14 of 235 patients with Parkinson's disease (5.69%), but not in 214 unrelated healthy controls. Multivariate analysis indicated the frequency of Gly2385Arg variant in the female patients with early age at onset is higher than their male counterparts. The founder haplotype analysis showed the variant carriers shared the same founder. Clinically, the LRRK2 Gly2385Arg carriers presented with classical Parkinson's disease symptoms. Our study indicates that the LRRK2 Gly2385Arg variant is a potential ethnic-specific genetic risk factor of Parkinson's disease within Chinese Han ethnicity.