Whole genome sequencing and analysis of Bacillus sp. TTMP2, a tetramethylpyrazine-producing bacterium.

BACKGROUND: Tetramethylpyrazine has been extensively studied as an anticancer substance and a flavor substance in the fields of medicine and food industry. A strain with high tetramethylpyrazine production was screened from the fermented grains of Danquan winery. Genome sequencing can reveal the potential roles of bacteria by thoroughly examining the connection between genes and phenotypes from a genomic perspective. METHODS AND RESULTS: In this study, whole genome of this strain was sequenced and analyzed. This paper summarized the genomic characteristics of strain TTMP2 and analyzed genes related to the synthesis of tetramethylpyrazine. Bacillus sp. TTMP2 has a complete metabolic pathway for acetoin and tetramethylpyrazine metabolism. Gene function was analyzed by COG annotation, GO annotation, KEGG annotation and functional annotations for lipoproteins, carbohydrate-active enzymes, and pathogen-host interactions. Phylogenetic analysis indicated that Bacillus velezensis had the high homology with Bacillus sp. TTMP2. Genomes of 16 Bacillus species cover all genes of Bacillus, suggesting that genus Bacillus has an open pan-genome and can survive in diverse environments. CONCLUSION: The analysis of genome sequencing data from Bacillus sp. TTMP2 showed that its metabolic characteristics could be deeply understood, indicating that this bacterium had a particular role in tetramethylpyrazine synthesis.

Causal relationship between asthma and inflammatory bowel disease : A two-sample bidirectional mendelian randomization analysis.

BACKGROUND: Based on the findings of current observational studies, asthma and inflammatory bowel disease (including Crohn's disease and ulcerative colitis) are associated; however, their causal association cannot be established due to methodological limitations. OBJECTIVES: we use two-sample bidirectional mendelian randomization (MR) to overcome the confounding factors and explore the causal link between asthma and inflammatory bowel disease. METHODS: After selecting asthma and IBD-related genome-wide association studies (GWAS) data and screening single nucleotide polymorphisms (SNPs), MR analysis was performed by four methods: inverse variance weighted (IVW), MR-Egger, maximum likelihood, and weighted median (WM), while Cochran's Q test was used to detect heterogeneity and MR-Egger intercept to detect horizontal pleiotropy. Finally, we used the leave-one-out method and funnel plot to perform sensitivity analysis. RESULTS: We screened 57, 59, and 60 SNPs in the association analysis of asthma and IBD, CD, and UC, respectively. The results of MR analysis showed that asthma only increased the risk of CD (IVW: OR = 1.1712, 95% CI = 1.0418-1.3167, P value = 0.0082; maximum likelihood: OR = 1.1739, 95% CI = 1.0428-1.3215, P value = 0.0080). Neither forward nor reverse MR analysis revealed heterogeneity or horizontal pleiotropy. Similarly, we did not find potential directional pleiotropy by funnel plot, and the leave-one-out method did not suggest a significant effect of a single SNP on the overall results. CONCLUSIONS: we found a negative correlation between asthma and Crohn's disease, but more research is needed to confirm this.

Optimization of Molasses and Soybean Meal Content to Enhance Tetramethylpyrazine Yield by Bacillus sp. TTMP20.

Microbial fermentation for the production of tetramethylpyrazine (TTMP) is considered to be the most promising method, and the development of a cheap fermentation substrate is of great importance for large-scale TTMP production. In this study, inexpensive by-products from the food industry, i.e., molasses and soybean meal (instead of glucose and tryptone), were used as substrates for TTMP fermentation. The pretreatment of soybean meal was explored in order to achieve a better fermentation effect. The contents of each component in the fermentation medium were optimized by central composite design (CCD). The optimum contents were as follows: 72.5 g/L of molasses, 37.4 g/L of diammonium hydrogen phosphate (DAP), 53.4 g/L of soybean meal, and 5 g/L of yeast powder. The software predicted a maximum TTMP yield of 1469.03 mg/L, and the actual TTMP yield was 1328.95 mg/L for the validation experiment in the optimum medium. Under the optimum conditions (72.5 g/L of molasses, 37.4 g/L of DAP, 53.4 g/L of soybean meal, and 5 g/L of yeast powder), the actual maximum TTMP yield (1328.95 mg/L) in this study was much higher than the TTMP yield (895.13 mg/L) under the conditions (150 g/L of molasses, 30 g/L of DAP, 30 g/L of tryptone, and 10 g/L of yeast powder) of our previous study published in Molecules. In this study, the TTMP yield improved by 48.46%, with decreased molasses (more than half), decreased yeast powder (half) and by-product soybean meal instead of tryptone compared to our previous study. In summary, the cheaper fermentation medium had a higher TTMP yield in this study, which improves the application potential of Bacillus sp. TTMP20.

Meta-linguistic awareness skills in Chinese-speaking children with hyperlexia: A single-case study.

Purpose: The aim of the study is to examine the meta-linguistic awareness skills contributing to reading aloud in a Chinese-speaking child with hyperlexia. Methods: Case study approach was used with one case of hyperlexia (TYH) and two control groups: typically developing (TD) children matched for chronological age (CA) and TD children matched for mental ability (MA). A battery of phonological, morphological, and orthographic awareness skill tests were administered. Results: Results from the modified t-test found that the hyperlexic child did not demonstrate advanced meta-linguistic awareness skills in comparison with the two control groups. On the contrary, TYH's morphological awareness skills were even lower than the CA control group. Also, in the orthographic awareness test, TYH demonstrated weaker knowledge of character structure and components than the two control groups although his ability in the recognition of real words is intact. In addition, the predictability of orthographic awareness skill was comparable to the CA group with predicted score showed no difference to his obtained score, while TYH achieved a significantly higher reading score than what his morphological awareness skills should predict with reference to TD children of similar age; as well as what his phonological awareness skill predict with reference to the MA group. Conclusion: The findings suggest that TYH can achieve advanced reading ability with comparable phonological and orthographic awareness skill, despite his weakness in morphological awareness. It is concluded that the hyperlexic reading in Chinese might be achieved through the direct mapping between the whole character and the sound.

Production of Tetramethylpyrazine from Cane Molasses by Bacillus sp. TTMP20.

2,3,5,6-Tetramethylpyrazine (TTMP) is an active ingredient of Ligusticum wallichii Franch. It can be used in medicine and food fields. In this study, Bacillus sp. TTMP20 was applied to produce TTMP using cane molasses as a carbon source. After pretreatment with phosphoric acid, 170 mL/L treated molasses, combined with 10 g/L yeast powder, 30 g/L tryptone and 30 g/L (NH4)2HPO4 were used for fermentation. After 36 h, TTMP output reached the highest value of 208.8 mg/L. The yield of TTMP using phosphoric acid-treated molasses as carbon source was 145.59% higher than control. Under the sulfuric acid treatment process of molasses (150 g), the maximum yield of TTMP was 895.13 mg/L, which was 183.18% higher than that of untreated molasses (316.1 mg/L). This study demonstrated that molasses is a high-quality and inexpensive carbon source for the manufacture of TTMP, laying the groundwork for the future industrial production of TTMP.

Optimization of Extraction Conditions of Carotenoids from Dunaliella parva by Response Surface Methodology.

Extraction conditions can exert a remarkable influence on extraction efficiency. The aim of this study was to improve the extraction efficiency of carotenoids from Dunaliella parva (D. parva). Dimethyl sulfoxide (DMSO) and 95% ethanol were used as the extraction solvents. The extraction time, extraction temperature and the proportions of mixed solvent were taken as influencing factors, and the experimental scheme was determined by Central Composite Design (CCD) of Design Expert 10.0.4.0 to optimize the extraction process of carotenoids from D. parva. The absorbance values of the extract at 665 nm, 649 nm and 480 nm were determined by a microplate spectrophotometer, and the extraction efficiency of carotenoids was calculated. Analyses of the model fitting degree, variance and interaction term 3D surface were performed by response surface analysis. The optimal extraction conditions were as follows: extraction time of 20 min, extraction temperature of 40 °C, and a mixed solvent ratio (DMSO: 95% ethanol) of 3.64:1. Under the optimal conditions, the actual extraction efficiency of carotenoids was 0.0464%, which was increased by 18.19% (the initial extraction efficiency of 0.03926%) with a lower extraction temperature (i.e., lower energy consumption) compared to the standard protocol.

An Analysis on Promoting Prefabrication Implementation in Construction Industry towards Sustainability.

As a game-changing technology with significant environmental, economic, and social benefits, prefabricated technology has attracted attention and has been increasingly adopted in the construction industry. Although multitudinous studies have investigated various aspects of prefabrication in construction, a thorough review of its current development state that synthesized environmental, economic, and social sustainability dimensions remains overdue. Therefore, this study aims to fill this research gap by constructing a systematic framework, analyzing the research status quos, and providing recommendations for future research. This study first conducted a holistic review of 768 references with NVivo. A research foci framework that represented the body of knowledge in prefabrication in construction was developed with five levels, which were advantages, hindrances, stakeholders, promotion policies, and strategy spectrum. Following the framework, the in-depth analyses from the perspectives of environmental, economic, social sustainability, technologies development, and promotion strategies were performed. The current research domains were further linked with potential research directions for promoting prefabricated construction towards sustainability. The study is of value in both offering references for policy formulation and stakeholder practice and providing recommendations for future research.

[Expression and Clinical Significance of CTGF in Patients with Multiple Myeloma].

OBJECTIVE: To detect the relationship between CTGF in the bone marrow of MM patients and osteolytic lesion of myeloma, moreover, to investigate the clinical significance of CTGF in MM. METHODS: Fifity-four MM patients treated in our hospital from March 2019 to April 2020 were enrolled, and 28 healthy volunteers were selected as the control group. The plasma in bone marrow of the patients was collected, and the ELISA was used to detect the level of CTGF in bone marrow plasma and the relationship between its and clinical characteristics were statistically analyzed. RESULTS: The CTGF level of MM patients was significantly higher than those in the healthy control group (P<0.001); the CTGF level in male patients was higher than that in female patients (P=0.007); the CTGF level in MM patients with osteolytic lesions was significantly higher than patients without osteolytic lesions and controls (P=0.007, P=0.001). The CTGF level in MM patients was positively correlated with the number of bone lesions (P<0.001, r=0.52). CTGF levels in patients with ≥3 bone lesions were significantly higher than those with <3 bone lesions and without bone lesions (P=0.014, P=0.002). ROC curve result showed that CTGF expression level shows a significant diagnostic value for MM bone disease (P<0.001). CONCLUSION: The abnormally high expression of CTGF level in MM patients is related to the degree of myelomas osteolytic lesions and can reflect the progress of MM.

Roles of strontium and hierarchy structure on the in vitro biological response and drug release mechanism of the strontium-substituted bioactive glass microspheres.

The strontium (Sr) substituted bioactive glasses (BGs) have inimitable advantages for bone generation, but the influences of Sr substitution on bioactivity and biocompatibility are still in debate. A brand novel porous microstructure of Sr-substituted BG microspheres was prepared by an electro-spraying technique combined with phase inversion, in which in vitro biological response (bioactivity, cell viability, alkaline phosphatase (ALP) activity and extracellular matrix (ECM) mineralization) and drug release were clarified in view of physical chemistry and ionic release behavior. The electro sprayed bioactive glass (ESBG) microspheres involved three characteristic pores: <100 nm, 100-1000 nm and >1000 nm. The Sr substitution on molar basis hindered the bioactivity of the samples in some extent but the cell behavior of the MC3T3-E1 cells was not significantly discouraged. The drug release profile was also controlled by amount of Sr substitution. However, the porous structure of the microspheres conferred improvement in bioactivity and provided a distinct three-stage drug release mode. Therefore, the Sr substituted ESBG microspheres may provide an effective way to deliver a steady supply of therapeutic ions and drugs in bone implantation patients.

Epidemiologic features and intervention effect of fall injury among rural school-aged children in southwest China: a short-term cohort study.

Falls are the top one type in all unintentional injuries. In this study, we aim to explore the epidemiological characteristics of falls and assess the intervention effect. Our research had interviewed 2854 rural children in southwest China. Then, we used School-Family-Individual (SFI) comprehensive education model to conduct an intervention among 1506 children and follow up them for one year. The changes in injury knowledge and incidence rate before and after intervention were compared. We found the fall injury was 37.32% (178/477) and ranked top one in the total injuries. After intervention, the children's fall-injuries-related knowledge was significantly increased by 15.29 percent (P < 0.001). While falls incidence significantly decreased after- intervention (6.24% vs. 3.93%; P < 0.001). From the results we concluded that the falls rate was high and was the prior reason of all injuries. SFI intervention model can effectively reduce the incidence of the fall injury.

Effect of a multi-level education intervention model on knowledge and attitudes of accidental injuries in rural children in Zunyi, Southwest China.

OBJECTIVE: To explore the effect of a school-family-individual (SFI) multi-level education intervention model on knowledge and attitudes about accidental injuries among school-aged children to improve injury prevention strategies and reduce the incidence of pediatric injuries. METHODS: The random sample of rural school-aged children were recruited by using a multistage, stratified, cluster sampling method in Zunyi, Southwest China from 2012 to 2014, and 2342 children were randomly divided into intervention and control groups. Then children answered a baseline survey to collect knowledge and attitude scores (KAS) of accidental injuries. In the intervention group, children, their parents/guardians and the school received a SFI multi-level education intervention, which included a children's injury-prevention poster at schools, an open letter about security instruction for parents/guardians and multiple-media health education (Microsoft PowerPoint lectures, videos, handbooks, etc.) to children. Children in the control group were given only handbook education. After 16 months, children answered a follow-up survey to collect data on accidental injury types and accidental injury-related KAS for comparing the intervention and control groups and baseline and follow-up data. RESULTS: The distribution of gender was not significantly different while age was different between the baseline and follow-up survey. At baseline, the mean KAS was lower for the intervention than control group (15.37 ± 3.40 and 18.35 ± 5.01; p < 0.001). At follow-up, the mean KAS was higher for the intervention than control group (21.16 ± 3.05 and 20.02 ± 3.40; p < 0.001). The increase in KAS in the intervention and control groups was significant (p < 0.001; KAS: 5.79 vs. 1.67) and suggested that children's injury-related KAS improved in the intervention group. Moreover, the KAS between the groups differed for most subtypes of incidental injuries (based on International Classification of Diseases 10, ICD-10) (p < 0.05). Before intervention, 350 children had reported their accident injury episodes, while after intervention 237 children had reported their accidental injury episodes in the follow-up survey. CONCLUSIONS: SFI multi-level education intervention could significantly increase KAS for accidental injuries, which should improve children's prevention-related knowledge and attitudes about such injuries. It should help children change their risk behaviors and reduce the incidence of accidental injuries. Our results highlight a new intervention model of injury prevention among school-aged children.

Features and risk factors of nonfatal injury among the rural children: a survey of seven schools in a mountain area in Southwest China.

OBJECTIVE: We aimed to investigate the patterns and risk factors of nonfatal injuries among rural mountain-area children in southwest China. METHODS: A stratified sampling method was used to recruit rural children aged 8 to 17 years (mainly 9-14 years) from 7 schools. Self-reported injuries during the past 12 months and relevant concerns were collected from June to December 2012 by using a structured questionnaire in a class interview. RESULTS: The mean age of the 2,854 children was 12.2 ± 1.5 years. The probability of annual injury was 16.7% (95% confidence interval [95% CI] 15.3-18.1%), with slightly higher injury risk for boys than girls (17.7% vs. 16.0%; P>0.05). The top 3 causes of injuries were falls (37.3%), animal-related incidents (20.6%), and burns (14.9%). The main injury risk factors included being involved in a violent episode (odds ratio [OR] 1.34, 95% CI 1.08-1.66, P = 0.007), maltreatment by parents or guardians (1.42, 1.17-1.72, P<0.001), and being from a single-child family (1.30, 1.10-1.66, P = 0.039). Older age was a protective factor (0.81, 0.76-0.87, P<0.001). CONCLUSIONS: The incidence of nonfatal injury among rural children was high, and falls were the leading cause. Younger children and boys from poor-care and poor-living environments were at increased risk of injury, which requires urgent attention. Injury prevention programs targeting these issues are needed in this mountain area and similar rural regions of China.

Influence of chronic intermittent hypoxia on growth associated protein 43 expression in the hippocampus of young rats.

This study aimed to explore the pathological change to hippocampal neurons and the expression of growth associated protein 43 in 21-day-old young rats following chronic intermittent hypoxia. Hematoxylin-eosin staining results showed varying degrees of degeneration and necrosis in hippocampal neurons depending on the modeling time. Immunohistochemistry revealed that growth associated protein 43 expression in young rats following chronic intermittent hypoxia decreased, but that levels were still higher than those of normal rats at each time point, especially 4 weeks after modeling. During 1-5 weeks after modeling, a slow growth in rat weight was observed. Experimental findings indicate that chronic intermittent hypoxia may induce growth dysfunction and necrosis of hippocampal neurons, as well as increase the expression of growth associated protein 43 in young rats.

Development of a gold nanoparticles based chemiluminescence imaging assay and its application.

In this paper, a novel gold nanoparticles based protein immobilization method was designed. Biocomposites of gold nanoparticles and proteins were successfully coated on poly(methyl methacrylate) (PMMA) plates and polystyrene microtiter plates. The proteins could be immobilized on solid materials with high density and better bioactivity. Based on above design, chemiluminescence (CL) imaging assay for determination of H(2)O(2) and recombinant human interleukin-6 (rHu IL-6) was developed. The linear range and the loading capability were greatly improved when compared with imaging assay performed with direct proteins immobilization. Under the selected experimental conditions, a linear relationship was obtained between the CL intensity and the concentration of H(2)O(2) in the range of 1.0 x 10(-6) to 1.0 x 10(-4) mol L(-1), and rHu IL-6 in the range of 2.0-312.0 pg mL(-1). The detection limits were 2 x 10(-7) mol L(-1) (3sigma) for H(2)O(2) and 0.5 pg mL(-1) for rHu IL-6 with relative standard deviation of 3.8% for 3.0 x 10(-5) mol L(-1) H(2)O(2), and 4.4% for 39.0 pg mL(-1) rHu IL-6. This method has been applied to the determination of rHu IL-6 in human serum with satisfactory results.

Chemiluminescent imaging analysis of interferon alpha in serum samples.

Enzyme-linked immunosorbent assay (ELISA), horseradish peroxidase (HRP)-catalyzed fluorescent reaction, and oxalate chemiluminescence imaging analysis have been combined to develop a sensitive, simple, and rapid method for analysis of interferon alpha (alpha-IFN) in human serum samples. A typical "sandwich type" immunoassay was used. Reaction of o-phenylenediamine (OPD) with hydrogen peroxide (H(2)O(2)), catalyzed by HRP, produced 2,3-diaminophenazine (PDA), which was detected by chemiluminescence imaging analysis with the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H(2)O(2)-glyoxaline-PDA chemiluminescent system. The TCPO chemiluminescent imaging system is more sensitive and the chemiluminescence quantum yield is at least five times higher than for the luminol-H(2)O(2)-HRP-PIP (p-iodophenol) chemiluminescent imaging system. The results showed there was a very good linear correlation between response and amount of alpha-IFN in the range 1.3-156.0 pg mL(-1) (R = 0.9991) and the detection limit was 0.8 pg mL(-1) (S/N=3). The relative standard deviation (n = 9) was 4.7%. The proposed method has been used for successful analysis of the amount of alpha-IFN in human serum. The results obtained compared well with those obtained by conventional colorimetric ELISA and luminol chemiluminescent ELISA.

The study of a chemiluminescence immunoassay using the peroxyoxalate chemiluminescent reaction and its application.

The paper presented a novel chemiluminescence (CL) immunoassay method, which combines the advantages of traditional enzyme-linked immunosorbent assays (ELISA) and bis (2,4,6-trichlorophenyl) oxalate (TCPO)-hydrogen peroxide CL detection system. A fluorescent product 2,3-diaminophenazine (DAPN) was produced by reaction between o-phenylenediamine (OPDA, 1,2-diaminobenzene) and H(2)O(2) catalyzed by horseradish peroxidase (HRP). DAPN was excited by the reactive intermediate of TCPO-H(2)O(2) chemiluminescent reaction, and led to CL. The dependence of the CL intensity on the concentrations of antigen was studied. As analytical application, the proposed method was used for determination of recombinant human interleukin 6 (rHu IL-6) and beta-human chorionic gonadotropin (beta-HCG). Under the selected experimental conditions, a linear relationship was obtained between the CL intensity and the concentration of rHu IL-6 in the range of 4.0-625.0pg/ml, and beta-HCG in the range of 12.5-400.0mIU/ml. The detection limits were 0.5pg/ml for rHu IL-6 and 3mIU/ml for beta-HCG with relative standard deviation of 2.3 for 78.0pg/ml rHu IL-6, and 3.9 for 50.0mIU/ml beta-HCG. This method has been applied to the determination of rHu IL-6 in human serum and beta-HCG in urine with satisfactory results.

Sensors based on galvanic cell generated electrochemiluminescence and its application.

In this paper, a novel electrochemiluminescence (ECL) imaging sensor array was developed for determination of hydrogen peroxide (H2O2), which was based on Cu/Zn alloy galvanic cell generated ECL. In alkaline solution, Cu/Zn galvanic cell was formed because of corrosion effect, the galvanic cell could supply stable potential for ECL generation of luminol, and the weak ECL emission could be enhanced by H(2)O(2). The galvanic cell sensor array was designed by putting Cu/Zn alloy in 96-well microtiter plates separately. The relative ECL intensity was proportional with the concentration of hydrogen peroxide in the range of 1.0 x 10(-6) to 1.0 x 10(-4) mol l(-1) and the detection limit was 3.0 x 10(-7) mol l(-1) (3sigma), the relative standard deviation (R.S.D.) for 11 parallel measurements of 1.0 x 10(-5)mol l(-1) H2O2 was 4.0%.

[Comparison of curative effect of arthroscopic and open surgeries for retrograde intramedullary interlocking nails for femoral supracondylar fracture].

OBJECTIVE: To compare the curative effect of arthroscopic and open surgeries for retrograde intramedullary interlocking nails(RIIN) in the treatment of femoral supracondylar fracture. METHODS: A total of 40 patients with femoral supracondylar fracture underwent either arthroscopic (n=20) or open surgeries (n=20) for rRIIN. RESULTS: The operation time, surgical blood loss and bone union time in arthroscopic group was less than that in the open surgery group (P<0.05). Follow-up of the patients for 10.3+/-3.6 months revealed excellent results in 15 cases, good in 3 cases, and acceptable in 1 case in the arthroscopic group, according to the Kolmert criteria, while excellent in 7, good in 14, acceptable in 4 and poor in 2 cases in the open surgery group, without statistically significances in the rate of good or excellent outcomes between the two groups (90.0% vs 75.0%, P=0.246). CONCLUSION: Compared with open surgery, RIIN shortens the operation time, reduces intraoperative blood loss and accelerates bone union, thus benefiting the recovery of knee joint functions.

A micro-chemiluminescence determination of cyanide in whole blood.

A reactant volume self-controlled micro-device was presented and applied to the flow injection chemiluminescence (CL) for determination of cyanide in whole blood. A mini distiller was fabricated for cyanide extraction from the blood samples with the extraction efficiencies of cyanide > or = 98%. A fluidic control platform with air driving was fabricated. The described system showed the features of easy fabrication, undiluted sample injection, safe analysis operation, and suitability for automatic cyanide analysis. The calibration curve showed linearity in the cyanide concentration range of 5.0 x 10(-7) to 5.0 x 10(-5) mol l(-1) with the detection limits (3sigma) of 2.3 x 10(-7) mol l(-1). CL peak-height precision was 1.9% R.S.D. (n = 11) at the 1.0 x 10(-6) mol l(-1) cyanide level. The new devices were applied to the analysis of cyanide in rabbit whole blood samples and the results agreed well with those obtained from official method.

An on-line galvanic cell-generated electrochemiluminescence and flow injection determination of calcium in milk and vegetables.

An on-line Ag/Al galvanic cell was studied and employed to generate electrochemiluminescence (ECL) of calcein blue. The potential of the galvanic cell could be adjusted by varying the components of flow reagent or by using different metals to substitute for Ag or Al. The reported cell exhibited perfect capability of supplying a stable potential for ECL generation. Because the weak ECL of calcein blue could be greatly sensitized in the presence of calcium in alkaline solution, calcium contents in milk and vegetable samples were assayed; the results were validated with ICP-AES method. The method gave linear results in 1.0 x 10(-4) mol L(-1) to 8.0 x 10(-6) mol L(-1) calcium concentration range and the 3(sigma)limit of detection was to be 2.0 x 10(-6) mol L(-1). Experiment results imply that this model of ECL detection could be applied for instrument miniaturization with easy fabrication.