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To elucidate the variations in fungal diversity and cultivable yeast in different types of Daqu, third-generation sequencing technology was employed to analyze the fungal communities in high-temperature and medium-high-temperature Daqu. Additionally, traditional microbial pure culture methods and molecular biology techniques were utilized to investigate the yeast resources within Daqu. Alpha diversity analysis revealed significant differences (P

In this study, medium-high temperature Daqu (MHTD), the primary saccharification and fermentation agent in the production of strong-aroma Baijiu, was investigated across different fermentation stages. Employing national standard methods, artificial intelligence sensing technology, and high-throughput sequencing techniques, a comprehensive analysis was conducted to investigate the physicochemical properties, flavor, taste, and changes in fungal community structure during the MHTD fermentation process. The correlation between quality indicators of MHTD and dominant fungal genera was also explored. The results revealed that, during fermentation, the moisture content, levels of organic sulfur compounds, terpenes, hydroxides, and astringency gradually decreased in MHTD, while saltiness steadily increased. The fungal community structure of MHTD was established in the early stages of fermentation. However, as fermentation progressed, fungal community diversity gradually decreased, accompanied by significant changes in the relative abundance of certain genera. Specifically, the relative abundance of Saccharomycopsis gradually decreased, while Thermoascus and Candida increased, becoming dominant in the later stages of fermentation. Procrustes analysis and correlation analyses indicated a significant correlation (P

In order to analyze the high-temperature Daqu bacterial taxa and their physicochemical properties in Linyi area, Shandong Province, Illumina MiSeq high-throughput sequencing technology and pure culture technology were used to analyze the bacterial taxa of high-temperature Daqu in Lanling wine, and then the physicochemical indexes were determined by conventional detection methods in this study. Finally, the correlation between bacterial taxa and physicochemical indexes in Daqu was explored. The results showed that the main genera of bacteria in Daqu were Kroppenstedtia, Thermoactinomyces, Saccharopolyspora, Staphylococcus, Rhodococcus, Weissella and Bacillus, et al. Bacillus subtilis, Bacillus licheniformis and Bacillus coagulans were cultureable dominant bacterial isolates. There was a large difference in saccharification, esterification, alcoholization and liquefaction between the 15 parts of high-temperature Daqu. Kroppenstedtia, Staphylococcus, Leucobacter, Rhodococcus and Weissella were significantly positively correlated with esterification, fermentation, alcoholization, esterification and liquefaction (P

In order to clarify the differences in the community structure and gene function of medium-high temperature Daqu bacteria in Linyi, Shandong Province and Zhoukou, Henan Province. MiSeq high-throughput sequencing technology was used to analyze the bacterial community structure of medium-high temperature Daqu in the two regions, and the potential functions of bacterial taxa were analyzed based on the clusters of orthologous groups of proteins (COG). The result of α diversity showed that there were no significant differences in richness and diversity (P>0.05). Compared with Daqu in Zhoukou area, at the phylum level, the average relative abundance of Actinobacteria and Bacteroidetes in Daqu in Linyi area were significantly lower (P

In order to analyze the correlation between fungus and physicochemical characteristics of medium-temperature Daqu in Xiangyang, Hubei Province, high-throughput sequencing and pure culture techniques were used to analyze the samples from Hubei Longzhong Wine Industry, and physicochemical and sensory indexes were evaluated, then the influence of fungus on its quality was revealed. The result showed that all the physicochemical indexes of Daqu samples met the requirements of QB/T 4259-2011 "Fragrant Daqu", and there were great differences in esterification and saccharification among samples (P

In this study, the physicochemical characteristics and bacterial taxa of 11 fen-flavor Daqu samples from Changzhi, Shanxi Province were analyzed using a combination of physicochemical analysis and MiSeq high-throughput sequencing, and the correlation between them was revealed. The physicochemical analysis showed that the variation coefficients of amino acid nitrogen, liquefaction power and alcoholization power of different fen-flavor Daqu samples were greater than 15%, with the largest difference in liquefaction power and the smallest difference in ash. MiSeq high-throughput sequencing analysis found that the bacterial genera presented in all samples with an average relative content >1.0% were Aquabacterium, Pelomonas, Bacillus, Ralstonia, Sphingomonas, Burkholderia, Methylobacterium and Bradyrhizobium, and a large number of bacterial taxa were shared among the different samples. Correlation analysis revealed that Sphingomonas showed a significant positive correlation with protein content (P

On the basis of using electronic tongue and electronic nose to evaluate the quality of Zhaguangjiao, MiSeq high-throughput sequencing technology was used to analyze the bacterial diversity and explore the effect of different salt concentration on its quality. The results showed that with the increase of salt concentration, the taste and flavor indexes changed regularly. At the gate level, Firmicutes (84.87%) gradually increased, while Proteobacteria (13.65%) decreased gradually. At the genus level, the relative content of Lactobacillus increased gradually, and the average relative content was as high as 60.20%. When the salt concentration was 5%, the flora structure of Zhaguangjiao tended to be stable, and strength of good indicators (umami) in flavor and taste was moderate, while the relative intensity of defective indicators (sourness) was generally low. Thus, it can be seen that adding 5% salt to the Zhaguangjiao can significantly improve the quality of the Zhaguangjiao.

In this study, to reveal the physicochemical properties and microbial taxa of Hong Qu produced in Shandong, China three Hong Qu samples were collected from a Shandong Qu-making company, and based on the determination of their physicochemical indexes and fermentation characteristics, the microbial taxa were resolved using MiSeq high-throughput sequencing technology, and the Bacillus and lactic acid bacteria contained in them were isolated and identified. The results showed that the moisture content of the three Hong Qu samples ranged from 8.27% to 10.30%, acidity ranged from 1.0 to 1.3 mmol/10 g, amino acid nitrogen content ranged from 131.65 to 172.76 mg/100 g, saccharification power ranged from 476 to 764 U/g, esterification power ranged from 331 to 548 U/g, and fermentation power, liquefaction power and alcoholization power were less than 10 U/g. The bacterial taxa of Hong Qu were mainly composed of Weissella (41.93%) and Acetobacter (9.99%), and the fungal taxa were mainly composed of Pichia (55.83%) and Monascus (21.24%), where the correlation between bacteria and alcoholization power was significant, and while the correlation between fungi and saccharification power was significant (P

In this study, Illumina MiSeq sequencing technology was used to analyze the bacterial community structure of Zha-Chili from Xingshan and Baojing. The α-diversity analysis showed that the community diversity and abundance of Zha-Chili in Xingshan were significantly lower than those in Baojing (P

To explore the effects of three pepper varieties on bacterial taxa and quality of Zhaguangjiao, Zhaguangjiao prepared using Capsicum frutescens Linn., Capsicum annuum L., and Capsicum annuum L. var. grossum (L.) Sendt. for the test group, while two control groups without the addition of any vegetables and with the addition of only Apium graveolens var. rapaceum were set up, and based on that, the quality of Zhaguangjiao was digitally evaluated and its bacterial taxa were resolved by high-throughput sequencing. Finally, the correlations between Zhaguangjiao quality and dominant bacterial genera were revealed. It was found that, compared with Zhaguangjiao prepared using Capsicum frutescens Linn., both the bacterial diversity and abundance of Zhaguangjiao prepared using Capsicum annuum L. and Capsicum annuum L. var. grossum (L.) Sendt. were higher. Zhaguangjiao prepared using Capsicum annuum L. and Capsicum annuum L. var. grossum (L.) Sendt. were dominated by Lactobacillus with average relative abundances of 77.53% and 94.41%, respectively, while Zhaguangjiao prepared using Capsicum frutescens Linn. was dominated by Pseudomonas with an average relative abundance of 67.35%. Compared with Zhaguangjiao prepared using Capsicum frutescens Linn., the sourness and content of aromatic compounds of Zhaguangjiao prepared using Capsicum frutescens Linn. was dominated by Pseudomonas were significantly higher, while the contents of organic sulfides, terpenes and nitrogen oxides among volatile substances were significantly lower. After Pearson correlation analysis, it was found that elevating the abundance of Lactobacillus in the fermentation system of Zhaguangjiao could obviously suppress the undesirable flavor and improve the content of aroma substances in Zhaguangjiao. Zhaguangjiao is popular for its sour and spicy taste, due to Capsicum annuum L. var. grossum (L.) Sendt. is less spicy, choosing Capsicum annuum L. as the main raw material to produce Zhaguangjiao has a positive effect enhancing the overall quality of the product.

In order to compare and study the quality difference of Suanjiangshui between purebred fermentation and starter fermentation, Lactobacillus fermentum pure-bred fermentation and old brine fermentation were used to make Suanjiangshui, and the sensory quality was detected by bionic equipment such as electronic tongue, electronic nose and colorimeter. The results showed that the umami of pure fermentation of Lactobacillus fermentum was obviously higher than that of fermentation with old brine, while the sourness and astringency showed opposite trend. In terms of volatile flavor, the intensity of aromatic substances in Suanjiangshui fermented by pure lactic acid bacteria was significantly higher than that fermented by starters (P

In order to characterize and identify the chemical components in different parts of Artemisia argyi(roots, stems, leaves, and seeds), compounds with antioxidant activity were screened. In this study, ultra-performance liquid chromatography-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt-quadrupole time-of-flight-tandem mass spectrometry(UPLC-ABTS-Q-TOF-MS) was used as an online combination technique. Poroshell 120 SB-Aq(3.0 mm×150 mm, 2.7 μm) was used as the column, and acetonitrile(A)-0.2% formic acid water(B) was adopted as the mobile phase to perform gradient elution and was scanned in positive and negative ion modes. MassLynx software was utilized, and combined with reference substances and related literature, the chemical components of different parts of A. argyi were identified and compared. The antioxidant active components were detected by using the online detection system, and the antioxidant activities of active components of different parts of A. argyi were compared and evaluated by scavenging efficiency. As a result, a total of 87 compounds were identified from extracts of different parts of A. argyi, and 38, 72, 85, and 33 components were identified from roots, stems, leaves, and seeds. 22 compounds with antioxidant activity were screened, and 14, 17, 20, and 11 compounds with antioxidant activity were identified from roots, stems, leaves, and seeds. The results show that there are certain differences in chemical components and antioxidant components of different parts of A. argyi, which provides data support for the resource utilization and further research and development of A. argyi.

Soil microorganisms dominate the biogeochemical cycles of elements in glacier forelands, which continue to expand due to the climate warming. We analyzed the soil microbial functional characteristics among three types of glacier forelands on the Tibetan Plateau: Yulong Glacier (Y), a temperate glacier; Tianshan Urumqi Glacier No.1 (T), a sub-continental glacier; and Laohugou Glacier No.12 (L), a continental glacier. Here, soil microbial functional genes were quantified using quantitative microbial element cycling technology (QMEC). We found that, in the three glacier forelands, the abundances of soil microbial functional genes related to hemicellulose degradation and reductive acetyl-CoA pathway were highest compared with other carbon-related functional genes. The main nitrogen cycling genes were involved in ammonification. The functional genes of the phosphorus cycle and sulfur cycle were related to organic phosphate mineralization and sulfur oxidation. Furthermore, the soils of the temperate glacier foreland with better hydrothermal conditions had the most complex microbial functional gene structure and the highest functional potentials, followed by those of the soils of continental glacier foreland with the driest environment. These significant differences in soil microbial functional genes among the three types of glacier forelands verified the impacts of geographic difference on microbial functional characteristics, as well as providing a basis for the study of soil microbial functions and biogeochemical cycles in glacier forelands.

Wetlands are an important global source and sink of methane. However, human activities and climatic conditions are causing serious degradation of wetlands in China. In response to this, the relevant departments have progressively carried out wetland restoration projects over the past few years. To investigate the response of microbial communities of bacteria, methanogens, and methanotrophs during degradation and restoration of wetlands, soil samples were collected from undegraded reed wetlands, degraded reed wetlands, and restored reed wetlands in the Songnen Plain. Microbial diversity and community composition were studied by high-throughput sequencing based on the 16S rRNA gene of bacteria, the mcrA gene of methanogens, and the pmoA gene of methanotrophs. The results indicate that the degradation of reed wetlands results in a decrease in bacterial and methanogenic α -diversity and an increase in methanotrophic α -diversity. Bacterial α -diversity and methanogenic α -diversity were both significantly positively correlated with soil water content. At different taxonomic levels, higher relative abundances of Rhizobiales and Methanobacteriaceae were detected in the undegraded wetland soils. Wetland degradation decreased the relative abundance of Rhizobiales but increased that of the pathogenic bacteria Burkholderiaceae and microorganisms resistant to harsh and extreme environments including Sphingomonas , Rubrobacter , Methylobacter , Methylomonas , and Methylococcus . In the restored wetland soils, the relative abundances of Bacillus , Methanosarcinaceae, Methanomicrobiaceae, and the type Ⅱ methanotroph Methylocystis were higher. Therefore, different wetland conditions can indirectly change soil properties and, consequently, change the community structure of methanogens and methanotrophs.

In this study, we investigated the effectiveness and microbial mechanism of Bacillus amyloliquefaciens biofertilizer on reducing ammonia volatilization in farmland soil. Pot experiments were carried out to explore the effects of B. amyloliquefaciens biofertilizer (BB) and chemical fertilizer on soil ammonia volatilization, crop yield and quality, and soil microbial community. Four fertilization strategies were tested, namely no fertilizer (CK), 100% chemical fertilizer (C), 50% BB and 50% chemical fertilizer (B1), and 100% BB (B2). The dynamic flow-through chamber method was used to determine the soil ammonia volatilization flux after fertilization. The soil bacterial community during the peak period of ammonia volatilization was analyzed using 16S rDNA high-throughput sequencing. The results showed that the amount of ammonia volatilization in B1 and B2 decreased by 79.5% and 84.8%, respectively, as compared with treatment C. B2 had the lowest nitrate content and the highest yield; the yield of B2 increased by 50.5% and 12.3% as compared to that of CK and C, respectively. B1 had the highest content of vitamin C, which was 67.6 mg ·kg -1 . The application of BB improved the diversity and richness of soil bacterial community, especially the relative abundance of Bacillus and Nitrospira . This shows that BB plays an important role in preventing air pollution and improving nitrogen utilization.

Glaciers are constantly retreating because of global warming. In this study, three soil samples along the forefield of Urumqi Glacier No. 1 were collected. The effects of warming on the microbial community in the glacier forefield were investigated through a 150-day laboratory experiment. In this experiment, two temperature treatments were performed at 5℃and 15℃. The results showed that with increasing deglaciation age, the concentrations of carbon and nitrogen increased and the abundance and alpha diversity of microbial communities increased in the original samples. The 150-day laboratory experiment indicated that warming insignificantly changed the copy number of archaea and bacteria. Furthermore, it changed the microbial community composition, and the changes varied in different sampling sites. Based on the analysis of abundant OTUs changing significantly with warming, the sampling sites with shorter deglaciation age had stronger response with warming, representing an increase in the abundance of genus Thiobacillus. Furthermore, these results revealed that warming caused different effects on microbes along glacier forefield and thus, it could provide important characteristics of the microbial community with warming in alpine glacier regions.

Excessive reclamation leads to rapid degradation of wetland ecosystems. Microbial changes in wetland soils under the influence of human activities can sensitively indicate degradation of soil quality and ecosystem functions. To study the effects of different land use patterns on microbial community structure of wetlands, the Sanjiang Wetland Protected Area of Fuyuan, Heilongjiang Province, was selected as the research area. Soil samples were collected from replanting legume crop area, rice wetland, and primitive peat wetland. Then, the bacterial community structure in the soil was investigated with high-throughput sequencing based on the 16S rRNA gene. The relationship between bacterial community and environmental factors was further explored. The results indicated that, based on the bacterial phylum, there are no significant differences between the microbial community structures of soils under different land use patterns. Nevertheless, at the genus level, higher abundance of Blastocatella , Coxiella , and Rickettsia were detected in the legume rhizospheric soil. In the paddy soil, the relative abundances of Massilia , Nitrosomonas , and Bradyrhizobium are higher, while in the peatand soil, the higher contents are of Rhizomicrobium , Arthrobacter , and Bacillus . The results of Chao1 and Shannon index indicate that the microbial diversity of the paddy soil was higher than in the legume rhizospheric soil and peatland soils. However, no significant differences on bacterial diversity between the legume rhizospheric soil and peatland soils were observed. The results of the correlation analysis indicate that soil reclamation triggers a shift in microbial community mainly because of its influence on soil pH, moisture, and nutrients.

Fine particles(PM 2.5 ) collected by two middle volume air samplers in the suburbs of Baoding and the urban area of Beijing during Jan 10 th to 17 th , 2015, were used to compare the community characteristics of cultivable bacteria in the two sites. In this study, we observed the particle morphology of PM 2.5 using a field emission scanning electron microscope (FESEM), analyzed the air mass transportation source of the two sampling sites by the NOAA/ARL HYSPLIT-4 backward trajectory model, and measured the concentrations of the major water-soluble ions and heavy metal elements in the PM 2.5 samples using ion chromatography, continuous flow analyzer and ICP-MS. The results indicated that, the cultivable bacteria of PM 2.5 contained three phyla, which were Firmicutes, Actinobacteria and α -Proteobacteria. Nine genera and 17 species of cultivable airborne bacteria were isolated and identified. The most abundant phylum was Firmicutes. The Gram-positive bacteria accounted for more than 90%. The spore-forming Bacillus which was the dominant species contributed 68.15% and 75% to the total bacteria in urban Beijing and the suburbs of Baoding, respectively. The difference in the community structure of PM 2.5 's cultivable bacteria in the two sampling areas may be affected by PM 2.5 's physical and chemical properties and air mass transportation.

Uncontrolled release and arbitrary irrigation reuse of rural wastewater may lead to water pollution, and the microbial pathogens could threaten the safety of freshwater resources and public health. To understand the microbial community structure of rural wastewater and provide the theory for microbial risk assessment of wastewater irrigation, microbial community diversities in the Membrane Bio-Reactor (MBR) process for rural wastewater treatment was studied by terminal restriction fragment length polymorphism (T-RFLP) and 16S rDNA gene clone library. Meanwhile, changes of Arcobacter spp. and total bacteria before and after treatment were detected through real-time quantitative PCR. The clone library results showed that there were 73 positive clones included Proteobacteria (91. 80%), Firmicutes (2. 70%), Bacteroidetes (1. 40%), and uncultured bacteria (4. 10%) in the untreated wastewater. The typical pathogenic genus Arcobacter belonging to e-Proteobacteria was the dominant component of the library, accounting for 68. 5% of all clones. The main groups and their abundance in different treatments were significantly distinct. The highest values of species abundance (S), Shannon-Wiener (H) and Evenness (E) were observed in the adjusting tank, which were 43. 0, 3. 56 and 0. 95, respectively. The real-time quantitative PCR results showed that the copy number of Arcobacter spp. was (1. 09 ± 0. 064 0) x 10(11) copies.L-1 in the untreated sewage, which was consistent with the result of 16S rDNA gene clone library. Compared to untreated wastewater, bacterial copy number in the treated effluent decreased 100 to 1 000 times, respectively, suggesting that MBR treatment system could remove the microbial quantity in such scale. In the recycled water, the physicochemical parameters and indicator bacteria met the water quality standard of farmland irrigation. However, further research is needed to estimate the potential health risks caused by residual pathogenic microorganisms in future.

The dynamics of rhizospheric fungal diversity and biomass at different sampling stages associated with two transgenic insectresistant cottons expressing Cry1Ac protein and their control varieties were studied under greenhouse conditions, followed by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real-time polymerase chain reaction (Q-PCR), in order to evaluate the ecological security of planting transgenic cotton expressing Cry1Ac protein. The results indicated that the fungal superior bands in rhizosphere of transgenic Bt cotton were similar with that of control cotton at four sampling stages, the more obvious difference in the blurred bands among transgenic Bt cotton, JM20 and SHIYUAN321 was detected. The rhizospheric fungal biomass of transgenic Bt cotton SGK321 was significantly lower than that of its parental control cotton at seedling stage, while the slight decrease in fungal biomass of transgenic Bt cotton XP188 was detected at boll forming stage, the ill-defined decrease, even growing tendency in two transgenic Bt cottons was detected at other stages. However, the difference of rhizospheric fungal community compositions and biomass was not only existed between transgenic cotton and its control, but also between SHIYUAN321 and JM20, and the same phenomenon was also detected between transgenic Bt cotton SGK321 and XP188. Hence, Bt protein is not the only incentive resulting in the difference in fungal community composition and diversity, the decrease in biomass between transgenic cotton and untransgenic cotton, different cotton varieties has an effect on them.

Blood stasis theory (BST) is widely used in the department of Chinese medical dermatology. Skin lesion we often see and modern medical examination results can be used as evidence for diagnosing BST and indications for using it. Better efficacy could also be obtained by using BST in treating wind evil or heat evil induced skin disease, and itching, hemorrhagic and stubborn dermatoses as well.

Cellulose was usually degraded by microbial communities in natural habitats. Construction of a simple cellulolytic consortium is necessary to understand the underlying interaction within microorganisms involved in cellulose conversion. A screening approach was developed to obtain a simple microbial community with the ability of cellulose degradation to methane. This technique was based on the method of enrichment culture accompanying with denaturing gel gradient electrophoresis (DGGE) fingerprint detection technology and roll-tube method. Moreover, a four-strain mixed culture capable of degrading cellulose to methane was isolated from Zoige alpine wetland of the Tibetan Plateau. The results showed that the microbial consortia consisted of three functional groups: the cellulolytic bacterium Clostridium glycolicum, the non-celluloytic bacteria group of Trichococcus flocculiformis and Parabacteroides merdae, and the methanogenic bacterium Methanobacterium subterraneum. This four-strain co-culture can convert cellulose to methane. In the future, the isolated cellulolytic consortia could provide a platform for controlling metabolic pathways and genetic modification involved in methane production from cellulose.

To prepare processed products with different methods, in order to study the impact of auxiliary materials and temperature on chemical components of Euphorbia ebracteolata, and establish specific chromatograms of different processed products. Wel-chorm-C18 column (4.6 mm x 250 mm, 5 microm) was used and eluted with a gradient program, with acetonitrile (A)-water(B). The column temperature was 25 degrees C, and the detection wave length was set at 226 nm. The aim was to determine the content of effective components in different processed products--ebracteolata cpd B, ebracteolata cpd C and jolkinolide B and establish respective characteristic fingerprints to compare with similarity. The results showed that the content of ebracteolata cpd B, ebracteolata cpd C first increased and then decreased with the rise in temperature. Different processed products showed significant difference in HPLC spectrograms, with a low similarity. This study showed great impacts of auxiliary materials and temperature on chemical components of E. ebracteolata. As the vinegar processing method had higher attenuation and and synergistic effects than other methods, the auxiliary material vinegar cannot be replaced by chemical reagent acetic acid.

Nitritation is an important part of the biological nitrogen removal process, and the performance of the process was determined by the microbial community structure. To explore the microbial adaptability to different sewage, the microbial diversity and the amount of bacteria were investigated in a high ammonia wastewater treatment process and a sewage treatment process using the clone library of bacterial 16S rDNA, the phospholipid fatty acid method (PLFA) and the quantitative PCR. The clone library results showed that there was a significantly difference in bacterial community structure of these two processes, although the dominant bacteria belong to the Proteobacteria and Bacteroidete, there were more clusters in the sewage treatment process. The PLFAs results showed that the microbial diversity index and the evenness index of the high ammonium wastewater treatment process were significantly low. The quantitative PCR results showed that amounts of ammonia oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) in the high ammonium wastewater treatment process were higher than these in sewage treatment process. The copy number of AOB was higher than the copy number of NOB in the high ammonia wastewater treatment process by three orders magnitude. The copy number of AOB was higher than the copy number of NOB in sewage treatment process by two orders of magnitude.

A microbial whole-cell biosensor CB10 for the bioavailability assessing of Cr6+ was constructed by molecular biotechnology. The regulatory gene and promoter of CB10 was from the chromium resistance system of plasmid pMOL28 from Cupriavidus metallidurans CH34, and the reporter gene of CB10 was luc which was derived from Photinus pyralis. Finally, its response characteristic was discussed under different incubation conditions e. g. pH and temperature. The results showed that a microbial whole-cell biosensor CB10 had been successfully constructed which could respond to Cr6+ within 30 min, with a LOD for Cr6+ of 2 micromol x L(-1). When the incubation concentration of Cr6+ was between 20 micromol x L(-1) and 200 micromol x L(-1), the luc activity of the CB10 biosensor was in linear correlation with the concentration of Cr6+. When the concentration of heavy metal was in the range of 10-50 micromol x L(-1), the response of CB10 was relatively more specific. Moreover, high concentrations of Pb2+, Mn2+ and Sb2+ could also induce CB10. By analyzing the response characteristic of CB10 biosensor, we could draw the conclusion that 15-30 degrees C and pH 4-7 were appropriate for CB10, and 30 degrees C and pH 7 were the optimal conditions for the incubation of the CB10 biosensor. The microbial whole-cell biosensor CB10 for the detection of Cr6+ was fast-responding, specific, sensitive and stable under various conditions. In prospective, it could be used in the fast detection of Cr6+ in water and assessment of the bioavailability of Cr6+ in soil.

A whole cell sensor, Pseudomonas fluorescens A506 (pTS), was immobilized by sodium alginate and the factors of cell density, immobilization time and beads usage were optimized. The performance of the immobilized cells was compared with that of the free cells. After 2 h immobilization,the increasing speed of fluorescent signal of immobilized cells was 2.26 times as high as that of the free cells,and the peak value was 2.23 times as high during the detection time ranging from 1.5 to 6.0 h. The constantly lower growth and density of the immobilized cell implied the enhanced signal intensity of single cells after immobilization. Meanwhile, the cell density decreased as the immobilization time prolonged. Cell density and immobilization time were the dominant factors affecting the detection signal. For benzene at higher concentrations, the immobilized biosensor showed more rapid signal response at the early period of detection.

Microbial whole-cell biosensor is an excellent tool to assess the bioavailability of heavy metal in soil and water. However, the traditional physicochemical instruments are applied to detect the total metal. Furthermore, microbial whole-cell biosensor is simple, rapid and economical in manipulating, and is thus a highly qualified candidate for emergency detection of pollution incidents. The biological component of microbial whole-cell biosensor mostly consists of metalloregulatory proteins and reporter genes. In detail, metalloregulatory proteins mainly include the MerR family, ArsR family and RS family, and reporter genes mainly include gfp, lux and luc. Metalloregulatory protein and reporter gene are related to the sensitivity, specificity and properties in monitoring. The bioavailability of heavy metals is alterable under different conditions, influenced by pH, chelate and detection methods and so on. Increasing the accumulation of intracellular heavy metal, modifying the metalloregulatory proteins and optimizing the detecting conditions are important for improving the sensitivity, specificity and accuracy of the microbial whole-cell biosensor. The future direction of microbial whole-cell biosensor is to realize the monitoring of pollutions in situ and on line.

Objective: To study the variation regularity of chemical constituents contained in Euphorbia ebracteolata after vinegar processing., Method: The colorimetric method was adopted for determining the variation of total lactone content in toxic constituents contained in E. ebracteolata decoction, with Kedde as the coloring reagent. The HPLC method was used for detecting the content variation of jolkinolide B and jolkinolide C, both were active constituents contained in E. ebracteolata decoction, before and after roasting with vinegar, in which Kromasil-ODS column (4.6 mm x 250 mm, 5 microm) was adopted, with the detection wavelength of 290 nm, column temperature at 25 degrees C , gradient elution with acetonitrile and water and the flow rate of 1.0 mL x min(-1)., Result: After roasting with vinegar, the total lactone content in E. ebracteolata was reduced from 0.60 to 0.45 mg x g(-1) , with active constituents, jolkinolide B and jolkinolide C, increased to varying degrees. The established chromatographic fingerprint contained risk information and could reflect the overall variation regularity of chemical constituents after roasting with vinegar., Conclusion: The chemical constituents of E. ebracteolata show significantly changes, especially a reduced toxicity, after roasting with vinegar. The increase in its efficacy may be related to the variation of these constituents.

Microbial communities of different depths (30, 60, 100, 150, 200cm) from the unexploited oilfield, gas field and control area were studied by PCR-DGGE and sequencing methods. The objectives of this study were to understand the microbial distribution in the regions of unexploited oil and gas fields, and to investigate the potential microbial indicators of oil and gas resources. The results showed that the Dice coefficients between different depths were very low (26-69.9). The microbial communities in the soil of 150 cm and 200 cm depth had greater richness (S > or = 19), diversity (H > or = 2.69) and evenness (E > or = 0. 90). The results of sequencing demonstrated that the bands from oilfield were mainly grouped into alpha-Proteobacteria, gamma-Proteobacteria, Actinobacteria, Acidobacteria with the predominance of gamma-Proteobacteria (75%). Most of the bands were related to oil-associated and hydrocarbon degrading bacteria, such as Methylophaga and Alcanivorax. While the gas field had alpha, beta, gamma, delta-Proteobacteria and Bacteroidetes, and gamma-Proteobacteria accounted for only 24%. More strains showed relativity to methanotrophs, such as Methylocystaceae. Thus, 150 cm and 200 cm were more suitable as the oil-gas exploration sampling depth. Methylocystaceae may act as potential indicators for gas resources, Methylophaga and Alcanivorax for oil.

The indigenous microbial communities within the plant phyllosphere are highly diverse and include many different species of bacteria, filamentous fungi, yeasts and algae which play important ecological roles. This study was the first attempt to assess the impact of abamectin treatments on microbial communities of broccoli phyllosphere using two culture-independent techniques of phospholipid fatty acid analysis (PLFA) and terminal restriction fragment length polymorphism (T-RFLP). Results showed that low concentration of abamectin treatments did not affect the microbial biomass and microbial community structure of broccoli phyllosphere significantly. However, high concentration of abamectin treatments significantly change the microbial community structure including a decrease of total and bacterial biomass, and a decrease in the ratio of Gram-positive bacteria to Gram-negative bacteria, but did not change the fungal biomass. Moreover, PLFA suggested that the number of unsaturated and cyclopropane phospholipid fatty acids (PLFAs: 16: 1omega9t, 18: 1omega7, cy17:0 and cy19:0) increased with high concentration abamectin treatment, while the saturated PLFAs i15:0, a15:0, i16:0 and a17:0 decreased. The appearance terminal restriction fragments (T-RFs: 58, 96, 236 and 420 bp) indicated that some bacteria might play a significant role in abamectin degradation in broccoli phylosphere, while the disappeared T-RFs (51, 89, 99, 338, 66, 223 and 482 bp) implied some other bacteria might potentially serve as microbial indicator of abamectin exposure.

The re-suspended road dust samples from 53 sites that could cover basically the entire Beijing in summer, and aerosol samples from three representative sites in Beijing in four seasons were collected to study concentrations and spatial variations of elements and ions in resuspended road dust and its contribution to the air-borne particulate pollution. Ca, S, Cu, Zn, Ni, Pb and Cd were main pollution elements, and Ca2+, SO4(2-), Cl-, K+, Na+ and NO3- were major ions in re-suspended road dust. Al, Ti, Sc, Co and Mg in re-suspended road dust were mainly originated from crustal source, while Cu, Zn, Ni and Pb were mainly originated from traffic emissions and coal burning, and Fe, Mn and Cd were mainly from industrial emissions and coal combustion. Ca2+ and SO4(2-) mainly came from construction activities, construction materials and secondary gas-particle conversions, Cl- and Na+ were derived from industrial wastewater disposal and chemical industrial emissions, and NO3- and K+ were from vehicle emissions, photochemical reactions of NO,, biomass and vegetable burning. The contribution of mineral aerosol from inside Beijing to the total mineral aerosols was 30% in spring of 2002, 70% in summer of 2002, 80% in autumn of 2003, 20% in winter of 2002. The pollution levels of some major pollution species, Ca, S, Cu, Zn, Ni, Pb, Fe, Mn, and Cd in resuspended road dust reached 76%, 87%, 75%, 80%, 82%, 90%, 45%, 51%, and 94%, respectively, and their contributions to the related elements in PM10 were 20%-45%, 5%-18%, 4%-50%, 2%-46%, 4%-52%, 5%-20%, 30%-60%, 20%-40% and 2%-25%. Re-suspended road dust from the traffic and construction activities is one of the major sources of the air-born particulate pollution in Beijing.

Aim: To investigate the action and mechanism of Syn-1A in reversing the activation of K(ATP) channel induced by weak acidic pH., Methods: The patches excised from Kir6.2/SUR2A expressing HEK-293 cells were used to establish inside-out configuration. To examine the actions of weak acidic pH in activation of the channel and the reverse action of Syn-1A on it, the inside-out patches were continuously perfused with the solution of pH from 7.4, 7.0, 6.8, 6.5 to 6.0 with or without Syn-1A. In vitro binding was employed to study the influence of different pH to the binding of Syn-1A to SUR2A subunit., Results: Syn-1A blocked pH 6.5, 6.8 and 7.0 induced activation of the channel, and Syn-1A binding to SUR2A were increased by reducing pH from 7.4 to 6.0., Conclusion: Syn-1A would assert some inhibition of the KATP channels, which might temper the fluctuation of acidic pH-induced K(ATP) channel opening that could induce fatal re-entrant arrhythmias.

A newly isolated Bacillus cereus strain HQ-1 was found to possess high capability to absorb silver ions. The study showed that the biosorption process could be described well by pseudo-second-order kinetic model and Freundlich isotherm model. Higher cell concentration was favorable to the biosorption. Temperature's effect on the biosorption was not obvious. The oxygenous and nitrogenous functional groups on the cell wall played a very important role in the process of biosorption. Microdeposits were formed by interaction between silver ions and biopolymers from the cell wall (such as polysaccharides, proteins and some hydrolysis).

To develop a new fusion protein (RGD)3/tTF for the therapy of the selective thrombosis of tumor blood vessels. The fused gene (RGD) 3/tTF was reconstructed by PCR, was cloned into vector pET22 b(+), and expressed in E. coli BL21 (DE3). The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was detected by clotting assay and F X activation assay. The specific binding of (RGD) 3/tTF to alphavbeta3 was analyzed by indirect ELISA. The recombinant plasmid pET22 b(+)/(RGD)3/tTF was obtained and expressed in E. coli BL21 (DE3). The purified fusion protein could induce blood coagulation, activiate F X. The ability of (RGD) 3/tTF binding specifically to alphavbeta3 was increased by 32%, compared with RGD/tTF. A new fusion protein (RGD) 3/tTF was successfully expressed in E. coli BL21 (DE3). The expressed proteins retained tTF activity and showed a higher binding to alphavbeta3 than that of RGD/tTF.

Aim: To study the cytotoxic effects of doxorubicin on apoptosis in glioma cell lines U343, U138, U373 induced by anti-human DR4/DR5 monoclonal antibodies (FMU1.4/FMU1.5) and the underlying mechanism., Methods: Expression of DR4/DR5 was quantitated by flow cytometry. Cytotoxicity exerted by FMU1.4/FMU1.5 on three cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis. The expression of cytochrome C, FLIP and Ca2+ concentration were also measured., Results: Following the treatment of doxorubicin DR4 and DR5 were highly expressed on the cell surface; The apoptosis of U138 and U373 induced by FMU1.4 and FMU1.5 was stronger. expression of cytochrome C and Ca2+ concentration were enhanced, whereas the expression of FLIP was downregulated., Conclusion: Subtoxic doxorubicin applied with antibodies caused higher cell death rate of glioma cells, which may be relevant to DR4/DR5, the release of cytochrome C and FLIP and Ca2+ concentration.

Aim: To construct the gene of humanized single-domain antibody hu3D3V(H) against human lung cancer, to express it in E.coli and analyze its activity., Methods: The mAb3D3V(H) was humanized by using CDRs grafting technique. The hu3D3V(H) gene was assembled by overlapping PCR. The expression vector pET22(b+)/hu3D3V(H) was constructed and transformed into E.coli BL21(DE3) and the hu3D3V(H) protein was expressed under IPTG induction. After purification by Ni(2+)-affinity chromatographic column, the activity of hu3D3V(H) protein was analyzed by indirect ELISA and competitive inhibition ELISA., Results: The target gene with correct sequence was obtained by overlapping PCR. The hu3D3V(H) protein was expressed as inclusion bodies with the yield of more than 30% of total bacterial proteins. After purification, the purity of hu3D3V(H) was more than 95%. The reactivity of purified protein was the same as parent antibody, and could inhibit the binding of mAb3D3 to L342 cells., Conclusion: The hu3D3V(H) still retains the reactivity and specificity of mAb3D3, which lays the foundation for its further clinical application.