Characterization, antioxidant activity, and mineral profiling of Auricularia cornea mushroom strains.

Background: Mushrooms are considered as next-generation healthy food components. Owing to their low-fat content, high-quality proteins, dietary fiber, and rich source of nutraceuticals. They are ideally preferred in formulation of low-caloric functional foods. In this view, the breeding strategies of mushroom Auricularia cornea (A. cornea) focusing on high yield and higher quality with rich nutritional values and health benefits are still needed. Materials and methods: A total of 50 strains of A. cornea were used to analyze the bio efficiency and the time required for fruiting body formation following the cultivation experiment. The calorimetric method was used to evaluate the antioxidant activity and quantify the crude polysaccharides and minerals content thereafter. Results: The results showed that the time required for fruiting body formation and biological efficiency varied significantly among the selected strains. Noticeably, the wild domesticated strain Ac13 of A. cornea mushroom showed the shortest fruit development time (80 days). Similarly, the hybrid strains including Ac3 and Ac15 possessed the highest biological efficiency (82.40 and 94.84%). Hybrid strains Ac18 (15.2%) and cultivated strains Ac33 (15.6%) showed the highest content of crude polysaccharides, while cultivated strains Ac1 and Ac33, demonstrated the highest content of total polysaccharides in the fruiting body (216 mg. g-1 and 200 mg. g-1). In the case of mineral content, the highest zinc contents were observed from the cultivated strain Ac46 (486.33 mg·kg-1). The maximum iron content was detected from the hybrid strain Ac3 (788 mg·kg-1), and the wild domesticated strain Ac28 (350 mg·kg-1). The crude polysaccharides of the A. cornea strain showed significant antioxidant potential, and the ability of Ac33 and Ac24 to scavenge DPPH radicals and ABTS, which was significantly improved compared to other strains, respectively. Principal component analysis was applied to examine the agronomic traits and chemical compounds of various strains of A. cornea mushrooms. The results revealed that cultivated, wild domesticated, and hybrid strains of A. cornea exhibited distinct characteristics in terms of growth, yield, and nutritional properties. Conclusion: The crude polysaccharides from A. cornea mushroom strains act as natural antioxidants, the wild, hybrid, and commercial A. cornea mushroom strains can achieve rapid growth, early maturation, and high yields. The evaluation of biochemical indexes and nutritional characteristics of strains with excellent traits provided a scientific basis for initiating high-quality breeding, provided germplasm resources for the production of "functional food" with real nutritional and health value.

Six-month repeated irradiation of 650 nm low-level red light reduces the risk of myopia in children: a randomized controlled trial.

PURPOSE: To evaluate whether the six-month repeated irradiation of 650 nm low-level red light (LLRL) decreases the risk of myopia onset in children. METHODS: This was a single-masked, randomized controlled trial. A total of 112 children (aged 6-12 years) were enrolled and randomized to the treatment group or control group in a 1:1 ratio. The cycloplegic spherical equivalent error (SER) of children at baseline was -0.5 diopter (D) to 3D. Children in the treatment group were irradiated with the 650 nm LLRL for 6 min daily. No intervention was given to the control. The primary outcomes are myopia incidence, change in cycloplegic SER, and change in axial length (AL). RESULTS: For the treatment group and control group, the six-month myopia incidence rates were 1.8% (95% confidence interval, CI: 0.2-4.9%) and 12.5% (95% CI: 5.5-21.9%), respectively. The difference was significant (p = 0.028). The median changes in AL for the treatment group and control group were -0.02 (interquartile range, IQR: -0.12 to 0.06) mm, and 0.09 (IQR: 0-0.18) mm, respectively. The difference was significant (p < 0.001). The median changes in cycloplegic SER for the treatment group and control group were 0 (IQR: 0-0.25) D, and -0.125 (IQR: -0.375 to 0) D, respectively. The difference was significant (p < 0.001). There was no adverse event. CONCLUSION: The repeated irradiation of 650 nm LLRL may have a strong effect for myopia prevention in children, without risk of adverse events. TRIAL REGISTRATION: this trial is retrospectively registered in the Chinese Clinical Trial Registry ( http://www.chictr.org.cn/ ), the registration number is ChiCTR2200058963.

Investigation of the Efficacy and Safety of 650 nm Low-Level Red Light for Myopia Control in Children: A Randomized Controlled Trial.

INTRODUCTION: To evaluate the 6-month efficacy and safety of 650 nm low-level red light (LLRL) for myopia control in children. METHODS: This was a single-center, single-masked randomized controlled trial. A total of 224 children aged 6-12 years with spherical equivalent error (SER) of - 6 diopter (D) to - 0.5 D were enrolled, and were randomized to LLRL group or control group. Children in the LLRL group underwent treatment twice daily, each lasting for 3 min, there was an interval of at least 4 h between treatments. Children in both groups were allowed to wear single-vision spectacles; no additional intervention was given to the control. The primary outcomes included change in cycloplegic SER and change in axial length (AL) during 6 months. RESULTS: The median 6-month changes in AL of the LLRL and control groups were - 0.06 mm (interquartile range, IQR - 0.15, 0) and 0.14 mm (IQR 0.07, 0.22), respectively. The difference between groups was significant (Z = 10.021, p < 0.001). The median 6-month changes in SER were 0.125 D (IQR 0, 0.375) and - 0.25 D (IQR - 0.5, 0) for the LLRL and control groups, respectively. The difference between groups was significant (Z = 8.827, p < 0.001). Compared with the control, the proportion of children with hyperopic shift in the LLRL group was higher (51.65% vs. 3.41%, p < 0.001), and the proportion of children with shortened AL in the LLRL group was higher (63.74% vs. 2.27%, p < 0.001). No adverse event was observed. CONCLUSION: 650 nm LLRL significantly slowed down the myopia progression in children aged 6-12 years, and there was no observable side effect in the short term.

RPA-Cas12aDS: A visual and fast molecular diagnostics platform based on RPA-CRISPR-Cas12a method for infectious bursal disease virus detection.

Infectious bursal disease (IBD), a major disease of birds, is caused by infectious bursal disease virus (IBDV). The disease can lead to immunosuppression, resulting in huge economic losses in the poultry industry. A specific, rapid, and simple detection method is important for the early diagnosis and prevention and control of IBDV. In this study, we established a naked-eye visual IBDV detection method, named "RPA-Cas12aDS", by combining recombinase polymerase amplification (RPA) with CRISPR-Cas12a-based nucleic acid detection. The detection process can be accomplished in 50 min, and uncapping contamination can be avoided. The detection results can be observed under blue or UV light. We used the RPA-Cas12aDS method to detect IBDV in bursa of Fabricius tissue samples of chickens, and the results were consistent with those obtained using commercial RT-PCR kits. This method presents great potential for visual, rapid, and point-of-care molecular diagnostics of IBDV in poultry.

Analysis of the Genome Sequence of Strain GiC-126 of Gloeostereum incarnatum with Genetic Linkage Map.

Gloeostereum incarnatum has edible and medicinal value and was first cultivated and domesticated in China. We sequenced the G. incarnatum monokaryotic strain GiC-126 on an Illumina HiSeq X Ten system and obtained a 34.52-Mb genome assembly sequence that encoded 16,895 predicted genes. We combined the GiC-126 genome with the published genome of G. incarnatum strain CCMJ2665 to construct a genetic linkage map (GiC-126 genome) that had 10 linkage groups (LGs), and the 15 assembly sequences of CCMJ2665 were integrated into 8 LGs. We identified 1912 simple sequence repeat (SSR) loci and detected 700 genes containing 768 SSRs in the genome; 65 and 100 of them were annotated with gene ontology (GO) terms and KEGG pathways, respectively. Carbohydrate-active enzymes (CAZymes) were identified in 20 fungal genomes and annotated; among them, 144 CAZymes were annotated in the GiC-126 genome. The A mating-type locus (MAT-A) of G. incarnatum was located on scaffold885 at 38.9 cM of LG1 and was flanked by two homeodomain (HD1) genes, mip and beta-fg. Fourteen segregation distortion markers were detected in the genetic linkage map, all of which were skewed toward the parent GiC-126. They formed three segregation distortion regions (SDR1-SDR3), and 22 predictive genes were found in scaffold1920 where three segregation distortion markers were located in SDR1. In this study, we corrected and updated the genomic information of G. incarnatum. Our results will provide a theoretical basis for fine gene mapping, functional gene cloning, and genetic breeding the follow-up of G. incarnatum.

Genetic linkage map construction and quantitative trait loci mapping of agronomic traits in Gloeostereum incarnatum.

Gloeostereum incarnatum is an edible medicinal mushroom widely grown in China. Using the whole genome of G. incarnatum, simple sequence repeat (SSR) markers were developed and synthetic primers were designed to construct its first genetic linkage map. The 1,048.6 cm map is composed of 10 linkage groups and contains 183 SSR markers. In total, 112 genome assembly sequences were anchored, representing 16.43 Mb and covering 46.41% of the genome. Selfing populations were used for quantitative trait loci (QTL) targeting, and the composite interval mapping method was used to co-localize the mycelium growth rate (potato dextrose agar and sawdust), growth period, yield and fruiting body length, and width and thickness. The 14 QTLs of agronomic traits had LOD values of 3.20-6.51 and contribution rates of 2.22-13.18%. No linkage relationship was found between the mycelium growth rate and the growth period, but a linkage relationship was observed among the length, width and thickness of the fruiting bodies. Using NCBI's BLAST alignment, the genomic sequences corresponding to the QTL regions were compared, and a TPR-like protein candidate gene was selected. Using whole-genome data, 138 candidate genes were found in four sequence fragments of two SSR markers located in the same scaffold. The genetic map and QTLs established in this study will aid in developing selective markers for agronomic traits and identifying corresponding genes, thereby providing a scientific basis for the further gene mapping of quantitative traits and the marker-assisted selection of functional genes in G. incarnatum breeding programs.

Diffusion Mechanism of Cinnamon Essential Oils Release from Calcium Alginate Based Controlled Release Films in Contact with Food Simulating Solvent.

Calcium alginate based controlled release films with moderate mechanical properties were fabricated in this paper. The diffusion mechanism of these films contacting food simulating solvent (FSS) was explored in some detail. With the increase of glycerol content, the diffusion coefficient (D) values of cinnamon essential oils (CEOs) diffusing to ethanol first increased slowly (0.3-0.6 mL), then vigorously (0.6-0.9 mL), and then mildly (0.9-1.2 mL). The D values of the CEOs diffused to water are all in the order of magnitude of 10-10 cm2/s. The D values of CEOs diffused from films EG3 and EGC1 to aqueous ethanol altered enormously at a small moisture percentage (w = 0.3), then continuously varied vigorously, and at last altered mildly in the range of w = 0.3-1. All the results above indicate that, considering the FSS, the diffusion ability of molecules is jointly determined by the size and distribution of free volume in the system (polymer + diffusive substance + solvents), the intermolecular interaction, and the partition coefficient of the solvents. In addition, several pairs of D values, such as DEG and DGA, are very close to each other, indicating that different kinds of interactions between different groups may have the same effect on the diffusion ability of molecules. The correlation between D1 and D2 indicates that polymeric emulsifier chains also exist in the polymer-rich layer. All the findings and analysis could provide the theoretical basis and data support for further molecular dynamic simulation and could guide the design of controlled release food packaging for food protection.

A Double-Switch Temperature-Sensitive Controlled Release Antioxidant Film Embedded with Lyophilized Nanoliposomes Encapsulating Rosemary Essential Oils for Solid Food.

In order to match the solid food oxidation during logistics and storage process under severe high temperature, a double-switch temperature-sensitive controlled release antioxidant film embedded with lyophilized nanoliposomes encapsulating rosemary essential oils (REOs) was prepared. The double switch temperature at 35.26 and 56.98 °C was achieved by development of a temperature sensitive polyurethane (TSPU) film. With biaxially oriented polyethylene terephthalate (BOPET) as a barrier layer, the intelligent complex film was prepared via coating the TSPU embedded with lyophilized nanoliposomes encapsulating REOs on BOPET. The results indicate that the REO is well encapsulated in nanoliposomes with encapsulation efficiency (EE) of 67.3%, high stability and lasting antioxidant effect during 60 days. The incorporation of lyophilized nanoliposomes containing REOs into TSPU remains the double-switch temperature-sensitive characteristic of the prepared TSPU. In agreement with porosity and WVTR results, the diffusion coefficient (D) of the antioxidant complex film sharply increases respectively at two switching temperatures, indicating that the intelligent double-switch temperature-sensitive controlled release property is functioning. Furthermore, compared with films directly added with REO, the lower Ds of films added with lyophilized nanoliposomes encapsulating REOs provides a longer-lasting antioxidant activity. Thus, the acquired controlled release antioxidant film sensitive to temperature at 39.56 and 56.00 °C can be potentially applied for protection of solid food during distribution and storage process under severe high temperatures.

Map-based cloning of genes encoding key enzymes for pigment synthesis in Auricularia cornea.

Color is an important quality attribute of fungi, and a useful marker for classification, genetic, and molecular research. However, there is much debate over which enzymes play key regulatory roles in pigment synthesis pathways among different fungi and even within the same species. Auricularia cornea is the most widely cultivated mushroom in the genus Auricularia; 1.834 million tons of this mushroom were produced in 2016 in China. Thus, systematic studies on its color inheritance and the genes encoding key enzymes for pigment synthesis have high scientific and economic value. In this study, the white strain ACW001 and the purple strain ACP004 of A. cornea were used as dikaryotic parents. Selfing populations of ACW001 and ACP004 were constructed with their monokaryotic strains. The fruiting body color of the two populations was consistent with that of their parents, confirming that the two parents were color homozygotes. All strains in the hybrid population of the two parents produced purple fruiting bodies. A robust hybrid strain (ACW001-33×ACP004-33) was selected from the hybrid population, and 87 monokaryotic strains of ACW001-33×ACP004-33 were obtained as a mapping population. Finally, a testcross population was constructed by crossing the mapping population with the test strain ACW001-9. The color genotype of each monokaryotic strain in the mapping population was identified by a fruiting test. The genomes of the two monokaryotic strains ACW001-33 and ACP004-33 were sequenced, and then simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) molecular marker primers were developed. Then, 88 pairs of primers that could distinguish the genotypes of the mapping population were used to construct a genetic linkage map. The genetic linkage map consisted of 12 linkage groups (LGs) spanning 1315.2 cM. The color control locus was preliminarily located at 24.5 cM of the 11th LG. Fine-mapping primers were designed based on sequence differences between ACW001-33 and ACP004-33 in the primary location region. Four color control candidate genes were located in an 8.2-kb region of ACW001-33_contig733 and a 9.2-kb region of ACP004-33_contig802. Homologous alignment and prediction of conserved domain analyses indicated that two of the color control candidate genes encoded proteins with unknown function, and the other two, ACP004_g11815 and ACP004_g11816, encoded glutamyl aminotransferases. These two genes were consecutively arranged on ACP004-33_contig802, and were likely to encode key enzymes in the γ-glutamine-4-hydroxy-benzoate (GHB) pigment synthesis pathway. Primers were designed from the flanking sequences of the two genes and used to analyze the testcross population. Products were amplified only from the 30 testcross strains with purple fruiting bodies, confirming the accuracy of the localization results. We discuss the deficiencies and advantages of map-based cloning in fungi vs. plants, and summarize the steps and requirements of the map-based cloning method for fungi. This study has provided novel ideas and methods for locating functional genes in fungi.

Development of a Molecular Marker for Fruiting Body Pattern in Auricularia auricula-judae.

The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then constructed two different near-isogenic pools. Then, we developed sequence characterized amplified region (SCAR) molecular markers associated with the fruiting body pattern based on sequence-related amplified polymorphism (SRAP) markers. Ten different bands (189-522 bp) were amplified using 153 pairs of SRAP primers. The SCAR marker "SCL-18" consisted of a single 522-bp band amplified from the cluster-type strains, but not the chrysanthemum strains. This SCAR marker was closely associated with the cluster-type fruiting body trait of A. auricula-judae. These results lay the foundation for further research to locate and clone genes controlling the fruiting body pattern of A. auricula-judae.

Construction of a genetic linkage map and QTL mapping of agronomic traits in Auricularia auricula-judae.

Auricularia auricula-judae is a traditional edible fungus that is cultivated widely in China. In this study, a genetic linkage map for A. auricula-judae was constructed using a mapping population consisting of 138 monokaryons derived from a hybrid strain (A119-5). The monokaryotic parent strains A14-5 and A18-119 were derived from two cultivated varieties, A14 (Qihei No. 1) and A18 (Qihei No. 2), respectively. In total, 130 simple sequence repeat markers were mapped. These markers were developed using the whole genome sequence of A. auricula-judae and amplified in A14-5, A18- 119, and the mapping population. The map consisted of 11 linkage groups (LGs) spanning 854 cM, with an average interval length of 6.57 cM. A testcross population was derived from crossing between the monokaryon A184-57 (from the wild strain A184 as a tester strain) and the mapping population. Important agronomic trait-related QTLs, including mycelium growth rate on potato dextrose agar for the mapping population, mycelium growth rate on potato dextrose agar and sawdust for the testcross population, growth period (days from inoculation to fruiting body harvesting), and yield for the testcross population, were identified using the composite interval mapping method. Six mycelium growth raterelated QTLs were identified on LG1 and LG4, two growth period-related QTLs were identified on LG2, and three yieldrelated QTLs were identified on LG2 and LG6. The results showed no linkage relationship between mycelium growth rate and growth period. The present study provides a foundation for locating genes for important agronomic characteristics in A. auricula-judae in the future.

Weibull kinetic modeling and nutritional effects of high-hydrostatic-pressure sterilization of soft-packing boiled bamboo shoots.

Application of high-hydrostatic-pressure (HHP) sterilization technology to boiled bamboo shoots was preliminarily discussed. An improved Weibull model that considered pressure and time as independent variables simultaneously for simulating the sterilization result was obtained by fitting the microbial lethal curves with high precision (R 2>0.98). HHP sterilization parameters could be calculated using this model with a certain disinfection rate. For a sterilization rate of 99.5%, treatment with a pressure of 400 MPa for 6 min or 500 MPa for 3 min at room temperature could exterminate all pathogenic microorganisms in boiled bamboo shoots, and control the total number of colonies at a low level below 100 CFU/g. Comparisons of the effects of HHP, high-pressure steam and microwave on the softness and contents of soluble protein and vitamin C of samples were also made. The results showed that the extent of damage of the products' original quality caused by HHP sterilization was less than that caused by the thermal and microwave treatment processes with equivalent sterilization percentages.

[Modeling of CO2 fluxes at cropland by using SiB3 model].

The aim is to study the influence of different vegetation types on CO2 fluxes at the same site, taking farmland as the object by using the simple biosphere model (SiB3) , select different vegetation types in Suzhou Dongshan site as input parameters of model to simulate CO2 fluxes, and compare with the observational data in Suzhou Dongshan site during 2011-04-16 to 2011-06-30. Results show that using corn as the site vegetation type input model to simulate CO2 fluxes showed a very good diurnal variation agreement compared to the measured data, but selecting the ordinary crop type input model to simulate CO2 fluxes, CO2 fluxes during the daytime in April and May were undervalued, and CO2 fluxes in June were overvalued; when we select the tea type to input the model, the simulated results significantly overestimated CO2 fluxes during the daytime in May and June. In addition, when using SiB3 model to simulate the daily CO2 fluxes, all three vegetation types input model can effectively simulate daily CO2 fluxes, but not significantly. It shows that choosing the right type vegetation input SiB3 model can effectively simulate the diurnal variation of CO2 fluxes, but can not effectively improve the daily CO2 fluxes simulation.

[Water intake of primary and middle school students in four cities of China].

OBJECTIVE: To investigate total intake of drinking water of primary and middle school students in four cities of China, and to provide scientific evidence for developing standard of intake of drinking water for primary and middle school students in China. METHODS: A total of 5914 primary and middle school students were selected from Beijing, Shanghai, Chengdu and Guangzhou using multi-stage random sampling method and 5868 subjects completed the survey. The information on the amounts of daily drinking water was recorded for seven consecutive days using a 24 hour measurement. The amounts of daily drinking water among different cities and ages and between boys and girls or urban and rural were analyzed. RESULTS: The average daily total drinking water of subjects was (1089 ± 540) ml, with significant differences among the four cities ((1165 ± 549), (1126 ± 544), (866 ± 421), (1185 ± 568) ml in Beijing, Shanghai, Chengdu and Guangzhou, respectively; F = 114.28, P < 0.05). The average daily drinking water was significantly higher in boys (1157 ± 575) ml than in girls (1026 ± 498) ml (Z = 8.88, P < 0.05), and in urban (1185 ± 578) ml than in rural (991 ± 480) ml (Z = 13.94, P < 0.05). The difference among different ages was statistically significant (F = 91.53, P < 0.05) with ascending order in 7 - 10, 11 - 13 and 14 - 17 years old ((953 ± 483), (1134 ± 551) and (1170 ± 557) ml respectively). CONCLUSION: The daily consumption of total drinking water is different in primary and middle school students among different cities, regions, genders and ages.

[Determination of heavy metals in ceramic food packaging containers by ICP-MS].

The direct determination method of heavy metals, Pb, Cd, Cr, Co, Ni, Zn, As and Sb in food packaging ceramics was studied by using inductive coupled plasma mass spectrometry (ICP-MS). Factors influencing the determination were also studied in detail. By choosing the optional operating parameters and appropriate isotopes of the element to be detected, the influence of mass spectrometry interference was conquered effectively. The detection limits were 0.023 microg x L(-1) for Pb, 0.004 microg x L(-1) for Cd, 0.030 microg x L(-1) for Cr, 0.025 microg x L(-1) for Ni, 0.012 microg x L(-1) for Sb, 0.180 microg x L(-1) for Zn, 0.017 microg x L(-1) for As, and 0.002 microg x L(-1) for Co. The recovery rates of the method were between 95.8% and 104.4%. The RSDs were between 1.1% and 3.3%. The method was simple, rapid, precise, convenient and suitable for daily inspection of import and export ceramic ware.

[Effects of applying endophytic fungi on the soil biological characteristics and enzyme activities under continuously cropped peanut].

A pot experiment was conducted to investigate the effects of applying endophytic fungi Phomopsis liquidambari strain B3, Phomopsis sp. strain NJ4.1, and Ceratobasidum stevensii strain B6 on the soil biological characteristics and enzyme activities under continuously cropped peanut at its different growth stages. Compared with the control, applying B3 increased the peanut yield significantly by 19.8%, and applying NJ4.1, B3 and B6 increased the peanut nodule number significantly by 20.4%, 29.3% and 27.6%, respectively. In the three treatments of applying endophytic fungi, the average population of soil bacteria and actinomycetes in the whole growth period of peanut was higher than that of the control, and the soil microbial biomass carbon was significantly greater at germination and seedling stages. The soil microbial biomass nitrogen increased at germination stage, but decreased at flowering stage. The DGGE analysis indicated that at flowering stage, the soil bacteria and fungi in treatment B3 had the largest band number and diversity. From germination stage to maturing stage, the three treatments of applying endophytic fungi had higher activities of soil invertase and catalase than the control, but less difference in soil urease activity. It was suggested that applying endophytic fungi could improve the peanut continuous cropping soil environment, and applying B3 had the best effect.

[Knowledge on drinking water of adults in four cities of China].

OBJECTIVE: To understand the status on knowledge of drinking water among adults aged 18 - 60 yrs in Beijing, Shanghai, Chengdu and Guangzhou of China. METHODS: A total of 1483 adults aged 18 - 60 yrs from the four cities (Beijing, Shanghai, Chengdu and Guangzhou) were selected using multiple-stage random sampling method. The information was collected by face-to-face interview. Through face to face interview, investigators surveyed the daily water intake, the harm of dehydration, time of drinking water and healthy drinks knowledge using a questionnaire. Knowledge awareness was compared through χ(2) test for different cities, urban and rural, genders. RESULTS: The unawareness rate of minimum drinking water volume (1200 ml/d) was 28.4% (421/1483). The rate in Chengdu was the highest (41.8%, 156/373). The rate in women (32.4%, 241/745) was significantly higher than that in men (24.4%, 180/738) (χ(2) = 11.55, P < 0.01). The unawareness rate of the harm of dehydration was 14.4% (213/1483). The rate in Chengdu (18.5%, 69/373) and Guangzhou (18.3%, 68/371) was higher than that in the other cities. The rate in rural (18.6%, 137/736) was significantly higher than that in urban (10.2%, 76/747) (χ(2) = 21.47, P < 0.01). The people who considered drinking water when they felt thirsty as healthy behavior accounted for 48.3% (716/1483). The rate in Chengdu was the highest (59.0%, 220/373). The unawareness rate of plain water being the best accounted for 18.9% (280/1483), while 17.9% (50/280) of them considered beverages as the best. CONCLUSIONS: The knowledge of drinking water among 18 - 60 yrs residents in the four cities is not comprehensive. We should enhance propaganda of healthy drinking water.

[Roles of PYK2 and caveolin-1 in alcoholic cardiomyopathy and the effect of drug intervention].

OBJECTIVE: To explore the protein expressions of PYK2 (protein-rich tyrosine kinase-2) and caveolin-1 in alcoholic cardiomyopathy (ACM) dog model and the effect of early drug intervention. METHODS: A total of 28 adult dogs were randomly divided into 4 groups: (i): alcohol-fed; (ii): alcohol plus valsartan, an angiotensin receptor blocker (ARB); (iii): alcohol plus carnitine; (iv): control group. At Month 6, the cardiac functions of all animals were evaluated by echocardiography. The concentrations of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in cardiac tissue were detected by chronometry. The protein expressions of collagen types I and III were determined by ELISA (enzyme-linked immunosorbent assay) while those of PYK2 and caveolin-1 evaluated by immunohistochemistry. The internal relationship of PYK2, caveolin-1, MDA and collagen types I & III was analyzed. RESULTS: Compared with the control group, the contents of SOD and GSH-Px significantly decreased in the alcohol, valsartan and carnitine groups while MDA significantly increased in the alcohol and carnitine groups. The relative contents in each group were as follows: MDA [alcohol: (28 ± 5) U/mg Pro; valsartan: (33 ± 13) U/mg Pro; carnitine: (33 ± 10) U/mg Pro], GSH-Px [alcohol: (188 ± 37) U/g Pro; valsartan: (362 ± 29) U/g Pro; carnitine: (282 ± 29) U/g Pro and MDA [alcohol: (19.4 ± 3.0) nmol/mg Pro; carnitine: (10.8 ± 2.1) nmol/mg Pro]. Compared with the control group, the concentration of collagen typeI significantly increased in the alcohol, valsartan and carnitine groups. In the alcohol-fed group, the protein expression of PYK2 gradually increased with the progression of ACM while that of caveolin-1 stayed at a low level. Through the interventions of valsartan and carnitine, the protein expression of PYK2 significantly increased while that of caveolin-1 significantly decreased (all P < 0.01). CONCLUSION: Alcohol promotes the myocardial oxidative stress and fibrosis through an elevated expression of PYK2 and a lowered expression of caveolin-1. Then a deterioration of cardiac structures and functions occur.

[Effect of valsartan and carnitine on cardiomyocyte Calpain-1 and Bcl-xl expressions of dogs with chronic alcohol intake-induced cardiomyopathy].

OBJECTIVE: To evaluate the effects of valsartan and carnitine on cardiomyocyte Calpain-1 and Bcl-xl expressions of dogs with chronic alcohol intake-induced cardiomyopathy. METHODS: Dogs were randomly assigned into 4 groups (n = 7 each): (1) alcohol fed (free access to 5%, 1(st) week; 10% 2(nd) week; 500 ml 25% bolus plus free access to 5% from 3 to 24 weeks, A); (2) alcohol + valsartan (5 mg×kg(-1)×d(-1), B); (3) alcohol + carnitine (300 mg×kg(-1)×d(-1), C); (4) Control (D). After six months, all animals were assessed for left ventricular (LV) function by echocardiography. The Bad and Bcl-xl protein expressions were evaluated by immunohistochemistry. The expression of Calpain-1 protein was determined with Western blot. Myocardial morphology was quantified on HE stained slices and under electron microscopy. The terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) was performed for apoptosis analysis. RESULTS: Compared with group D, LVEDD and LVESD were significantly increased while EF and FS significantly decreased in group A. In alcohol fed group, expressions of Bad and Calpain-1 protein were significantly increased while Bcl-xl protein expression was downregulated, all changes could be significantly attenuated by intervention with valsartan and carnitine (all P < 0.05). CONCLUSION: These data suggest that alcohol could promote cardiac myocyte apoptosis, reduce cardiac function and aggravate myocardial remodeling which valsartan and carnitine could reduce alcoholic cardiomyopathy by downregulating Calpain-1 and Bad protein expression and upregulating expression of Bcl-xl protein.

Involvement of semicarbazide-sensitive amine oxidase-mediated deamination in lipopolysaccharide-induced pulmonary inflammation.

Semicarbazide-sensitive amine oxidase (SSAO) resides on the vascular endothelium and smooth muscle cell surface and is capable of deaminating short chain aliphatic amines and producing toxic aldehydes and hydrogen peroxide. The enzyme, also known as a vascular adhesion protein-1, is involved in the inflammation process. This intriguing protein with dual functions is increased in the serum of diabetic and heart failure patients. In the present study we assessed the involvement of SSAO in a lipopolysaccharide-induced pulmonary inflammation model using transgenic mice that overexpress human vascular adhesion protein-1. Overexpression of SSAO activity increased the formation of protein-formaldehyde deposits in tissues. Lysine residues of proteins were the primary targets for cross-linkage with formaldehyde derived from deamination of methylamine. Lipo-polysaccharide-induced increases in inflammatory cells in the bronchoalveolar lavage (BAL) fluid were significantly higher in the transgenic than in the nontransgenic mice. BAL cell counts were also higher in the untreated transgenic than in nontransgenic mice. Blocking SSAO activity with a selective inhibitor significantly reduced the number of neutrophils as well as levels of macrophage inflammatory protein-1alpha, granulocyte colony-stimulating factor, tumor necrosis factor-alpha, and interleukin-6 in the BAL fluid. Inhalation of methylamine also increased BAL neutrophil counts. Together, these results suggest a role for SSAO-mediated deamination in pulmonary inflammation.