Appraising the Effect of Potential Risk Factors on Thyroid Cancer: A Mendelian Randomization Study.

CONTEXT: Various risk factors have been associated with the risk of thyroid cancer in observational studies. However, the causality of the risk factors is not clear given the susceptibility of confounding and reverse causation. OBJECTIVE: A 2-sample Mendelian randomization approach was used to estimate the effect of potential risk factors on thyroid cancer risk. METHODS: Genetic instruments to proxy 55 risk factors were identified by genome-wide association studies (GWAS). Associations of these genetic variants with thyroid cancer risk were estimated in GWAS of the FinnGen Study (989 cases and 217 803 controls). A Bonferroni-corrected threshold of P = 9.09 × 10-4 was considered significant, and P < 0.05 was considered to be suggestive of an association. RESULTS: Telomere length was significantly associated with increased thyroid cancer risk after correction for multiple testing (OR 4.68; 95% CI, 2.35-9.31; P = 1.12 × 10-5). Suggestive associations with increased risk were noted for waist-to-hip ratio (OR 1.85; 95% CI, 1.02-3.35; P = 0.042) and diastolic blood pressure (OR 1.60; 95% CI, 1.08-2.38; P = 0.019). Suggestive associations were noted between hemoglobin A1c (HbA1c) (OR 0.20; 95% CI, 0.05-0.82; P = 0.025) and decreased risk of thyroid cancer. Risk of thyroid cancer was not associated with sex hormones and reproduction, developmental and growth, lipids, diet and lifestyle, or inflammatory factors (All P > 0.05). CONCLUSION: Our study identified several potential targets for primary prevention of thyroid cancer, including central obesity, diastolic blood pressure, HbA1c, and telomere length, which should inform public health policy.

Exploration of interaction between angiotensin I-converting enzyme (ACE) and the inhibitory peptide from Wakame (Undaria pinnatifida).

The interaction between angiotensin I-converting enzyme (ACE) and the inhibitory peptide KNFL from Wakame was explored using isothermal titration calorimetry, multiple spectroscopic techniques and molecular dynamics simulations, and an inhibition model was established based on free energy binding theory. The experiments revealed that the binding of KNFL to ACE was a spontaneous exothermic process driven by enthalpy and entropy and occurred via multiple binding sites to form stable complexes. The complexes may be formed through multiple steps of inducing fit and conformational selection. The peptide KNFL had a fluorescence quenching effect on ACE and its addition not only affected the microenvironment around the ACE Trp and Tyr residues, but also increased the diameter and altered the conformation of ACE. This study should prove useful for improving our understanding of the mechanism of ACE inhibitory peptides.

Correction to: Effects of long-term straw incorporation on lignin accumulation and its association with bacterial laccase-like genes in arable soils.

The first funding No. should be 41671298 instead of 41301298.

Effects of long-term straw incorporation on lignin accumulation and its association with bacterial laccase-like genes in arable soils.

In this study, we aimed to investigate lignin accumulation and its relationship with the composition of bacterial laccase-like genes in three arable lands (i.e., upland limestone soil (UL), upland red soil (UR), and upland-paddy rotation red soil (UPR)), which are subjected to long-term straw incorporation. After 9-13 years of straw incorporation, the lignin content significantly increased from 337.1, 414.5, and 201.6 mg/kg soil to 2096.5, 2092.4, and 1972.2 mg/kg soil in UL, UR, and UPR, respectively. The dominant lignin monomer changed from vanillyl (V)-type to cinnamyl (C)-type in UR. Both V- and C-types were the dominant monomers in UPR, and V-type monomer remained the dominant monomer in UL. Compared with the treatment without straw, straw incorporation significantly promoted the activity of laccase enzyme and the abundance of bacterial laccase-like genes in all soils. The redundancy analysis showed that the main influencing factors on lignin accumulation patterns with straw incorporation were the laccase enzyme activity, nitrogen availability, and some specific bacterial communities possessing the laccase-like genes (e.g., Thermotogae and Acidobacteria). The variation partitioning analysis confirmed that the strongest influencing factor on lignin accumulation was the composition of bacterial laccase-like genes (explained 31.4% of variance). The present study provides novel insights into the importance of bacterial laccase-like genes in shaping lignin monomer accumulation with straw incorporation in arable soils.

Lignin and cellulose dynamics with straw incorporation in two contrasting cropping soils.

Incorporation of crop residues is essential to enhance soil organic matter in arable ecosystems. Here, we monitored the dynamics of cellulose and lignin, the most abundant constituents of plant residues, and their relationships with enzyme activities, microbial gene abundances and soil properties after 13-year long-term and one-year short-term crop straw incorporation into upland and upland-paddy soils in a field-based experiment. Lignin, rather than cellulose, accumulated in both soils following straw incorporation. Cellulose was almost completely converted into non-cellulose forms within 6 and 3 months after straw incorporation into upland and upland-paddy rotation soils, respectively. Whereas, lignin accumulated at the rate of 129 and 137 mg kg-1 yr-1 within 13 years' straw incorporation in upland and upland-paddy rotation, respectively. The predominance of recalcitrant vanillyl monomers in upland-paddy rotation indicated a high stability of lignin. Structural equation models revealed that the key factor driving cellulose and lignin dynamics was available nitrogen, followed by enzymes activities (cellobiohydrolases and laccases) and functional genes abundances (cbhI and laccase-like) as mediated by soil pH. Our findings highlighted that upland might have higher carbon sequestration rate, whereas upland-paddy rotation system was more beneficial for accumulation of recalcitrant organic fractions under crop residue incorporation.

Bacterial microbiota of Kazakhstan cheese revealed by single molecule real time (SMRT) sequencing and its comparison with Belgian, Kalmykian and Italian artisanal cheeses.

BACKGROUND: In Kazakhstan, traditional artisanal cheeses have a long history and are widely consumed. The unique characteristics of local artisanal cheeses are almost completely preserved. However, their microbial communities have rarely been reported. The current study firstly generated the Single Molecule, Real-Time (SMRT) sequencing bacterial diversity profiles of 6 traditional artisanal cheese samples of Kazakhstan origin, followed by comparatively analyzed the microbiota composition between the current dataset and those from cheeses originated from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy. RESULTS: Across the Kazakhstan cheese samples, a total of 238 bacterial species belonging to 14 phyla and 140 genera were identified. Lactococcus lactis (28.93%), Lactobacillus helveticus (26.43%), Streptococcus thermophilus (12.18%) and Lactobacillus delbrueckii (12.15%) were the dominant bacterial species for these samples. To further evaluate the cheese bacterial diversity of Kazakhstan cheeses in comparison with those from other geographic origins, 16S rRNA datasets of 36 artisanal cheeses from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy were retrieved from public databases. The cheese bacterial microbiota communities were largely different across sample origins. By principal coordinate analysis (PCoA) and multivariate analysis of variance (MANOVA), the structure of the Kazakhstan artisanal cheese samples was found to be different from those of the other geographic origins. Furthermore, the redundancy analysis (RDA) identified 16 bacterial OTUs as the key variables responsible for such microbiota structural difference. CONCLUSION: Our results together suggest that the diversity of bacterial communities in different groups is stratified by geographic region. This study does not only provide novel information on the bacterial microbiota of traditional artisanal cheese of Kazakhstan at species level, but also interesting insights into the bacterial diversity of artisanal cheeses of various geographical origins.

Effect of feeding Lactobacillus plantarum P-8 on the faecal microbiota of broiler chickens exposed to lincomycin.

Poultry and eggs are nutritious and healthy foods that contain high-quality proteins and low levels of fat compared with other meats. Recent studies have shown that poultry performance is closely associated with colonic health. Antibiotics are commonly used in the poultry industry to control diseases and enhance survival rates. However, antibiotic use can also result in host gut dysbiosis and immune dysregulation. Such imbalances compromise poultry health and growth performance. Thus, our study investigated the effect of Lactobacillus (L.) plantarum P-8 on the gut microbiome of chickens co-inoculated with the antibiotic lincomycin. Principal coordinate analysis showed that L. plantarum P-8 treatment shifted the faecal bacterial population structure. At the metagenomic level, the Clusters of Orthologous Groups (COGs) functional categories of P, C, N and A were overrepresented in the probiotic group. Additionally, the relative gene abundances of metabolic pathways involved in flagellar assembly, bacterial chemotaxis, nitrogen metabolism, sulfur metabolism, cofactor and vitamin biosynthesis were also higher in the probiotic group than the control; in contrast genes related to galactose degradation, carbon fixation, multiple sugar transport systems and ribosomes were underrepresented in the probiotic group. Our data suggest that feeding L. plantarum P-8 has the potential to improve metabolic activity and nutrient utilization of poultry. Furthermore, the faecal antibiotic resistomes of the two groups could be separated using principal components analysis, indicating that the probiotic treatment may modulate the intestinal antibiotic resistance gene pool by changing the population structure of the gut microbiota. This study has provided interesting insights into the application of probiotics in the poultry industry.

Variations in the patterns of soil organic carbon mineralization and microbial communities in response to exogenous application of rice straw and calcium carbonate.

The addition of exogenous inorganic carbon (CaCO3) and organic carbon has an important influence on soil organic carbon (SOC) mineralization in karst soil, but the microbial mechanisms underlying the SOC priming effect are poorly understood. We conducted a 100-day incubation experiment involving four treatments of the calcareous soil in southwestern China's karst region: control, (14)C-labeled rice straw addition, (14)C-labeled CaCO3 addition, and a combination of (14)C-labeled rice straw and CaCO3. Changes in soil microbial communities were characterized using denaturing gradient gel electrophoresis with polymerase chain reaction (PCR-DGGE) and real-time quantitative PCR (q-PCR). Both (14)C-rice straw and Ca(14)CO3 addition stimulated SOC mineralization, suggesting that organic and inorganic C affected SOC stability. Addition of straw alone had no significant effect on bacterial diversity; however, when the straw was added in combination with calcium carbonate, it had an inhibitory effect on bacterial and fungal diversity. At the beginning of the experimental period, exogenous additives increased bacterial abundance, although at the end of the 100-day incubation bacterial community abundance had gradually declined. Incubation time, exogenous input, and their interaction significantly affected SOC mineralization (in terms of priming and the cumulative amount of mineralization), microbial biomass carbon (MBC), and microbial community abundance and diversity. Moreover, the key factors influencing SOC mineralization were MBC, bacterial diversity, and soil pH. Overall, these findings support the view that inorganic C is involved in soil C turnover with the participation of soil microbial communities, promoting soil C cycling in the karst region.

Laccase activity is proportional to the abundance of bacterial laccase-like genes in soil from subtropical arable land.

Laccase enzymes produced by both soil bacteria and fungi play important roles in refractory organic matter turnover in terrestrial ecosystems. We investigated the abundance and diversity of fungal laccase genes and bacterial laccase-like genes in soil from subtropical arable lands, and identified which microbial group was associated with laccase activity. Compared with fungal laccase genes, the bacterial laccase-like genes had greater abundance, richness and Shannon-Wiener diversity. More importantly, laccase activity can be explained almost exclusively by the bacterial laccase-like genes, and their abundance had significant linear relationship with laccase activity. Thus, bacterial laccase-like gene has great potential to be used as a sensitive indicator of laccase enzyme for refractory organic matter turnover in subtropical arable lands.

Microbial Pollution Tracking of Dairy Farm with a Combined PCR-DGGE and qPCR Approach.

Animal husbandry is a traditional industry with regional characteristic in the Inner Mongolia of China. Recent years, animal breeding has been one of the main pollution sources in this area, followed by domestic sewage and industrial wastewater. The pollution of livestock farm feces may accelerate the development of pathogens and antibiotic resistance genes which pose health risks to humans and animals. In present research, culture-independent molecular ecological methods based on DGGE combined with qPCR were used to investigate the pollution to surrounding environment with different degrees of livestock farm. The cluster analysis of DGGE patterns showed that the livestock farm feces from point pollution source flowed with wastewater discharge has resulted in an impacted range of at least 3000 m, but it did not cause pollution to residential water delivered from upstream of sewage drain outlet. qPCR results revealed that 5 common pathogens (Escherichia coli, Enterococcus, Staphylococcus aureus, Shigella, and Salmonella) presented decreased trend as the sampled distance from point pollution source increased. Also, qPCR assays of 10 common antibiotic resistance genes (tetO, tetL, rpp, rpoB, sul2, sulA, floR, yidY, mphA, and ermC) which cause resistance to tetracycline, rifampicin, fluoroquinolone, quinolone, and erythromycin have been found in the environmental samples. This study clearly indicates the livestock farm discharge pollutants contaminated to the surrounding environment. Our data have provided important information to pollution control in the future.

[Effect of long-term fertilization on lignin accumulation in typical subtropical upland soil].

To investigate the effect of long-term fertilization on lignin accumulation and clarify its influencing factors in subtropical agricultural upland soils, alkaline CuO oxidation and gas chromatography was performed to quantify the amount of lignin and its monomers components (V, S and C). The soil samples were collected from the fertilization treatments of NPK and NPKS (NPK combined with straw) in Huanjiang County, Guangxi Province (limestone soil) and Taoyuan County, Hunan Province (red soil). The results showed that NPK had no significant effect on the lignin content (Sumvsc) of limestone soil, whereas the content in red soil significantly increased by (55 ± 1)%. For the NPKS treatment, the lignin content in limestone and red soil increased by (328 ± 4)% and (456 ± 9)%, respectively. After the same fertilization treatment, the proportion of cinnamyl (C)-type significantly increased in red soil, while a significant increase of vanillyl (V)-type monomers occurred in limestone soil, indicating that lignin degradation in agricultural soils was monomer specific. Furthermore, the acid-to-aldehyde ratios of syringyl-type [(Ac/Al)] or vanillyl-type [(Ac/Al)v] monomers tended to decrease after long-term fertilization with the higher value for limestone soil, suggesting the degree of lignin degradation in limestone was higher than that in red soil. Soil organic matter and total nitrogen were not correlated with lignin content, but were significantly correlated with the composition of VSC monomers. Meanwhile, the available nutrient content in the soil (available nitrogen, phosphorus, and potassium) was closely related to the contents and components of V, S, and C-type monomers (P<0.05). It indicated that the availability of soil nutrition should be considered as a key factor for the accumulation of lignin.

[Effects of Slope Position and Soil Horizon on Soil Microbial Biomass and Abundance in Karst Primary Forest of Southwest China].

To explore the effects of slope position and soil horizon on soil microbial biomass and abundance, chloroform fumigation extraction methods and real-time fluorescence-based quantitative PCR (Real-time PCR) were adopted to quantify the changes of soil microbial biomass C, N and abundance of bacteria and fungi, respectively. Soil samples were harvested from three horizons along profile, i. e., leaching horizon (A, 0-10 cm), transitional horizon (AB, 30-50 cm) and alluvial horizon (B, 70-100 cm), which were collected from the upper, middle and lower slope positions of a karst primary forest ecosystem. The results showed that slope position, soil horizon and their interaction significantly influenced the soil microbial biomass and abundance (P < 0.05). Different from A horizon, where SMBC was greater in lower than in upper slope position (P < 0.05), SMBC in AB and B horizons were highest in middle slope position. Similarly, SMBN was greater in lower than in upper slope position for A, AB and B horizons. Besides soil bacterial abundance in B horizon and fungal abundance in AB layer, the middle slope position had the highest value for all the three soil horizons (P < 0.05). Stepwise regression analysis showed that soil organic carbon, available nitrogen and pH were the key factors responsible for SMBC and SMBN variation, respectively, while the important factors responsible for the variation of bacteria abundance were available nitrogen and available phosphorus, and that for fungi abundance variation were available potassium.

[Basidiomycetous laccase gene diversity in two subtropical forest soils].

As one of the key enzymes involved in lignin decomposition of forest litter, laccase plays an important role in the carbon cycling in forest ecosystem. By using TA cloning and sequencing, a comparative study was conducted on the basidiomycetous laccase gene diversity in the O horizon (litter layer) and A horizon (surface soil layer, 0-20 cm) in two subtropical forests (a primeval evergreen deciduous broadleaved mixed forest and an artificial masson pine forest). For the same soil horizons, the basidiomycetous laccase gene diversity and richness were higher in the primeval forest than in the masson pine forest; for the same forest ecosystems, the basidiomycetous laccase gene diversity and richness in the primeval forest were slightly higher in O horizon than in A horizon, but those in the masson pine forest were apparently lower in O horizon than in A horizon. The two forest soils had the same dominant laccase gene-containing basidiomycetous populations, and most of the populations had high similarity of amino acid sequence to Mycena sp. or Pleurotus sp. belonging to Agaricales. Comparing with the A horizon in primeval forest and the O horizon in masson pine forest, the O horizon in primeval forest and the A horizon in masson pine forest had a relatively uniform distribution of basidiomycetous populations. The nucleotide sequence similarity of basidiomycetous laccase gene between the O and A horizons in the masson pine forest was higher than that in the primeval forest. This study showed that vegetation and soil horizon had significant effects on the basidiomycetous laccase gene diversity and community structure, and the discrepancies in the substrate availability for basidiomycetes and in the soil pH induced by the vegetation and soil horizon could be the driving forces.

Multicellular genesis of leaf primordium was demonstrated via chimaeric transgenic plant of maize (Zea mays L.) regenerated from Type II calli.

Type-II embryonic calli were induced from immature embryos of maize (Zea mays L.) genotype YD and bombarded with beta-glucuronidase gene. Bombarded calli were proliferated on normal N6 medium for 2 weeks at 26°C in the dark and selected on N6 medium containing 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 mg/l phosphinothricin (PPT) but without casamino acids and proline under the same conditions for 14 days. Regeneration was carried out on hormone-free MS medium containing 5 mg/l phosphinothricin at 26°C under 3000 lux illumination. Plants over 8 cm were transplanted into soil and sprayed with 250 mg/l phosphinothricin when two new leaves appeared. Except normal transgenic plants, chimaeric transgenics also were regenerated in the present work. The expression pattern of beta-glucuronidase gene in leaves of chimaeric transgenic plant revealed that more than one cell formed leaf primordium at the initial stage, and filial cells stemed from each cell in leaf primordium arranged in a row longitudinally from leaf base to leaf apex. There was a clear boundary as a straight line between the area formed by transformed cells and the area formed by normal cells. A hypothesis was put forward that the primitive cells in leaf primordium divided in a longitudinal style, resulted in leaf elongation, then the filial cells divided transversally and synchronously toward the outside to broaden the leaf.