Inhibitory Potential of Bifidobacterium longum FB1-1 Cell-Free Supernatant against Carbapenem-Resistant Klebsiella pneumoniae Drug Resistance Spread.

The widespread dissemination of carbapenem-resistant Klebsiella pneumoniae (CRKP) and its drug resistance transfer poses a global public health threat. While previous studies outlined CRKP's drug resistance mechanism, there is limited research on strategies inhibiting CRKP drug resistance spread. This study investigates the potential of Bifidobacterium longum (B. longum) FB1-1, a probiotic, in curbing the spread of drug resistance among CRKP by evaluating its cell-free supernatant (CFS) for antibacterial activity. Evaluating the inhibitory effect of FB1-1 CFS on CRKP drug resistance spread involved analyzing its impact on drug resistance and virulence gene expression; drug resistance plasmid transfer FB1-1 CFS exhibited an MIC range of 125 µL/mL against CRKP. After eight hours of co-culture, CFS achieved a 96% and 100% sterilization rate at two and four times the MIC, respectively. At sub-inhibitory concentrations (1/2× MIC), FB1-1 CFS reduced the expression of the bla_KPC gene, which is pivotal for carbapenem resistance, by up to 62.13% across different CRKP strains. Additionally, it markedly suppressed the expression of the uge gene, a key virulence factor, by up to 91%, and the fim_H gene, essential for bacterial adhesion, by up to 53.4%. Our study primarily focuses on determining the inhibitory effect of FB1-1 CFS on CRKP strains harboring the bla_KPC gene, which is a critical resistance determinant in CRKP. Furthermore, FB1-1 CFS demonstrated the ability to inhibit the transfer of drug resistance plasmids among CRKP strains, thus limiting the horizontal spread of resistance genes. This study highlights FB1-1 CFS's inhibitory effect on CRKP drug resistance spread, particularly in strains carrying the bla_KPC gene, thus offering a novel idea and theoretical foundation for developing antibacterial drugs targeting CRKP resistance.

Current Advances in Structure-Function Relationships and Dose-Dependent Effects of Human Milk Oligosaccharides.

HMOs (human milk oligosaccharides) are the third most important nutrient in breast milk. As complex glycans, HMOs play an important role in regulating neonatal intestinal immunity, resisting viral and bacterial infections, displaying anti-inflammatory characteristics, and promoting brain development. Although there have been some previous reports of HMOs, a detailed literature review summarizing the structure-activity relationships and dose-dependent effects of HMOs is lacking. Hence, after introducing the structures and synthetic pathways of HMOs, this review summarizes and categorizes identified structure-function relationships of HMOs. Differential mechanisms of different structural HMOs utilization by microorganisms are summarized. This review also emphasizes the recent advances in the interactions between different health benefits and the variance of dosage effect based on in vitro cell tests, animal experiments, and human intervention studies. The potential relationships between the chemical structure, the dosage selection, and the physiological properties of HMOs as functional foods are vital for further understanding of HMOs and their future applications.

Unusual (2R,6R)-bicyclo[3.1.1]heptane ring construction in fungal α-trans-bergamotene biosynthesis.

Bergamotenes are bicyclo[3.1.1]heptane sesquiterpenes found abundantly in plants and fungi. Known bergamotene derivatives all possess (2S,6S)-bergamotene backbone. In this study, two (+)-α-trans-bergamotene derivatives (1 and 2) with unusual (2R,6R) configuration were isolated and elucidated from marine fungus Nectria sp. HLS206. The first (+)-α-trans-bergamotene synthase NsBERS was characterized using genome mining and heterologous expression-based strategies. Based on homology search, we characterized another (+)-α-trans-bergamotene synthase LsBERS from Lachnellula suecica and an (+)-α-bisabolol synthase BcBOS from Botrytis cinerea. We proposed that the cyclization mechanism of (+)-α-trans-bergamotene involved endo-anti cyclization of left-handed helix farnesyl pyrophosphate by (6R)-bisabolyl cation, which was supported by molecular docking. The biosynthesis-based volatiles (3-6) produced by heterologous fungal expression systems elicited significant electroantennographic responses of Helicoverpa armigera and Spodoptera frugiperda, respectively, suggesting their potential in biocontrol of these pests. This work enriches diversity of sesquiterpenoids and fungal sesquiterpene synthases, providing insight into the enzymatic mechanism of formation of enantiomeric sesquiterpenes.

[Problems and strategy of further development of Chinese medicinal materials with edible values].

"Medicine and food homology" is a precious health care concept rooted in the culture of Chinese medicine and plays an important role in the development of the national health industry. It is consistent with the current global trend that food and medicine are mutually penetrating. Accordingly, the Chinese medicinal materials with edible values have an increasing production scale. Especially, in the context that the development of Chinese medicinal materials is switching from pursuing quantity to quality, the food field has become the main market for the new production capacity of Chinese medicinal materials, which has presented a broad prospect. However, the quality standards of raw materials, production methods, and administration ways vary between the materials for edible and medicinal purposes. Specifically, the food for medicinal use on the market cannot meet the quality standards of medicinal mate-rials, while the medicinal materials fail to meet the taste requirements as food. As a result, these problems cause difficulties in market circulation and supervision. In this paper, we analyzed the formation of Chinese medicinal materials with edible values, compared the food with medicinal value, common food, and functional food, and analyzed the different quality requirements of Chinese medicinal materials used in different scenarios. Further, we advised the differential development of Chinese medicinal materials in different directions(edible or medicinal use) from production to supervision. Including:(1) In the variety registration of Chinese medicinal materials with edible values, the variety breeding direction should be announced according to the requirements that medicinal materials care more about the content of active ingredients and food use materials preferentially need to meet the requirements of edible palatability.(2) Differentiation can be reflected in the selection of cultivation mode and planting and processing technology of medicinal materials, The differential production technical specification of medicinal materials with edible values should be developed. Such as the "simulated cultivation" mode is encouraged in the plant of medicinal materials to ensure its quality and the strict management of inputs and sufficient cultivation years should be guaranteed. While for edible medicinal materials, more kinds of cultivation techniques can be selected according to their processing methods.(3) The market supervision of medicinal materials with edible values should be guided by the purpose of their sales and use, which depends on the accurately recognize of the relationship between the properties of medicinal materials with edible values and the situation of pharmacopeia collection.(4) During publicity, when used as ordinary food and health food medi-cinal materials, it should be noticed that the publicity of the product's efficacy must meet the requirements of corresponding regulations.

[Cloning and functional verification of carboxyl CoA ligases(AeCCLs) in Arnebia euchroma].

Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.

[Analysis of economic benefits of Chinese medicine eco-agriculture based on multiple stakeholders].

As the most advanced environment-friendly production model in the international society, ecological agriculture of Chinese materia medica(CMM) is the only way for the development of modern agriculture. With the proposal of the declaration on ecolo-gical agriculture of CMM, "Don't grab land from farmland, don't be enemies of grass and insects, don't be afraid of barren slopes and forests, and live up to the green and green mountains", the ecological planting of CMM has blossomed all over the country, and formed a scientific theory, technology and model. Based on the theory and method of economics, this paper expounds the comprehensive benefits and development advantages of ecological agriculture of CMM from the perspectives of farmers(producers), patients(consumers) and the country. From the perspective of medicinal farmers, the input and output income of conventional agriculture and ecological agriculture of CMM such as Panax ginseng, Astragalus propinquus, Atractylodes lancea, and Bupleurum chinense were compared, and it was found that ecological agriculture of CMM had obvious advantages in net income, average annual income and input-output ratio, which could better promote farmers' income. From the perspective of patients, according to the same dose, the content of active ingredients in ecologically planted CMMs is significantly higher than that in conventionally-planted herbs, and the amount of effective substances taken by patients is also higher, so as to achieve better therapeutic effect. At the national level, ecological planting of CMM is the key to ensuring the high-quality development of CMM industry, increasing farmers' income, ensuring the safety of people's drug use and promoting the sustainable development of agriculture. It is also an important part of realizing the harmonious development of economy, society and environment and promoting ecological civilization. In general, the declaration on ecological agriculture of CMM embo-dies the core characteristics and goals of ecological agriculture, and also points of the path and vision of ecological agriculture of CMM in the future. The declaration will guide production practice, promote the benefit of farmers, and lay the foundation for the sustainable development of CMM industry.

[Effect of combined application of inorganic and organic fertilizers on growth and quality of Salvia miltiorrhiza].

The study is aimed through field experiments to study the effect of combined application of organic and chemical fertilizers on the growth and quality of Salvia miltiorrhiza, provide ideas for reducing fertilization while increasing the efficiency as well as improving the quality of produces. The experiment included 6 treatments viz., no fertilization(CK), full application of chemical fertilizer(F), 25% orga-nic fertilizer with 75% chemical fertilizer(M25), 50% organic fertilizer with 50% chemical fertilizer(M50), 75% organic fertilizer with 25% chemical fertilizer(M75), and fully apply organic fertilizer(M100). The results showed that:(1)from the perspective of yield and economic benefits, M75 was the best and M100 second;(2)for effective components, the combined application of organic and chemical fertilizers increased the content of main water-soluble components and the total content of effective components, among which M25 and M50 were better.

[Cloning and functional analysis of caffeic acid and rosmarinic acid glycosyltransferases from Arnebia euchroma].

Caffeic acid and its oligomers are the main water-soluble active constituents of the traditional Chinese medicine(TCM) Arnebiae Radix. These compounds possess multiple biological activities such as antimicrobial, antioxidant, cardiovascular protective, liver protective, anti-liver fibrosis, antiviral and anticancer activities. The phenylpropanoid pathway in plants is responsible for the biosynthesis of caffeic acid and its oligomers. Glycosylation can change phenylpropanoid solubility, stability and toxic potential, as well as influencing compartmentalization and biological activity. In view of the important role played by de-glycosylation in the regulation of phenylpropanoid homeostasis, the biosynthesis of caffeic acid and its oligomers are supposed to be under the control of relative UDP-glycosyltransferases(UGTs). Through the data mining of Arnebia euchroma transcriptome, we cloned 15 full-length putative UGT genes. After recombinant expression using the prokaryotic system, the crude enzyme solution of the putative UGTs was examined for the glycosylation activities towards caffeic acid and rosmarinic acid in vitro. AeUGT_01, AeUGT_02, AeUGT_03, AeUGT_04 and AeUGT_10 were able to glycosylate caffeic acid and/or rosmarinic acid resulting in different mono-and/or di-glycosylated products in the UPLC-MS analyses. The characterized UGTs were distantly related to each other and divided into different clades of the phylogenetic tree. Based on the observation that each characterized UGT exhibited substrate or catalytic similarity with the members in their own clade, we supposed the glycosylation abilities towards caffeic acid and/or rosmarinic acid were evolved independently in different clades. The identification of caffeic acid and rosmarinic acid UGTs from A. euchroma could lead to deeper understanding of the caffeic acid oligomers biosynthesis and its regulation. Furthermore, these UGTs might be used for regiospecific glycosylation of caffeic acid and rosmarinic acid to produce bioactive compounds for potential therapeutic applications.

[Establishment of RNA interfered hairy root system of two CYP450 genes in Arnebia euchroma and its influence].

In this study, based on the transcriptome database of suspension cells of Arnebia euchroma, we explored two candidate cytochrome P450 enzyme genes that might relate to the shikonin biosynthesis downstream pathway when CYP76B74 sequence was referenced. We constructed interference-type hairy roots of candidate genes and cultured them. We measured the fresh weight, dry weight, total naphthoquinone content, shikonin and its derivatives content and expression levels of key enzyme genes involved in shikonin biosynthesis pathway. The effects of candidate genes on the growth and shikonin production of A. euchroma hairy roots were discussed, and the possible regulatory mechanisms that candidate genes affected shikonin synthesis were discussed. Through local Blast and phylogenetic analysis, two candidate CYP450 genes(CYP76B75 and CYP76B100) with high homology to CYP76B74 in A. euchroma were screened, and corresponding interference hairy roots were constructed. Compared with the control(RNAi-control), the fresh weight of CYP76B75 interfered hairy root(RNAi-CYP76B75) and CYP76B100 interfered hairy root(RNAi-CYP76B100) were significantly reduced, while dry weight were not affected, so the dry rate increased significantly. Except for ß-acetoxyisovalerylalkannin, which is high in three groups of hairy roots, the contents of shikonin, deoxyshikonin, acetylshikonin, ß,ß'-dimethacrylicalkannin, ß-hydroxyisovalerylshikonin,ß-hydroxyisovalerylshikonin, isobutyrylshikonin and total naphthoquinones showed a consistent pattern: RNAi-CYP76B75>RNAi-CYP76B100>RNAi-control. Among them, the synthesis of ß-hydroxyisovalerylshikonin was most significantly promoted by interfering with the expression of CYP76B75. The content of ß-hydroxyisovalerylshikonin in RNAi-CYP76B75 was 11.7 times that of RNAi-control. RESULTS:: of real-time qPCR analysis showed that compared to RNAi-control, the expression levels of AePGT gene in RNAi-CYP76B75 and RNAi-CYP76B100 were not changed significantly, and the expression levels of CYP76B74 and AeHMGR were up-regulated. In addition, the expression level of CYP76B100 in RNAi-CYP76B75 was down-regulated, whereas in RNAi-CYP76B100, the expression of CYP76B75 was significantly up-regulated. Therefore, this study confirmed that when the expression of CYP76B75 and CYP76B100 were interrupted, the growth of hairy roots were suppressed, but the synthesis of shikonin were promoted. They might increase the shikonin biosynthesis by up-regulating the expression of CYP76B74 in the hairy roots of A. euchroma.

[Strategies for medicinal plants adapting environmental stress and "simulative habitat cultivation" of Dao-di herbs].

This paper analyzed life form, habitats and environmental stresses of medicinal plants and algal fungi collected in Chinese Pharmacopoeia(2015). ①It was found that only 0.94% of the medicinal plants mainly cultivated in field. The most common habitats of medicinal plants are divided into two types: those whose natural habitats are forest margins/undergrowth(about 42.53%) and those whose natural habitats are roadside, hillside, wasteland/sand(about 43.78%). The former mainly faces environmental stresses such as weak light, pests and diseases; the latter often faces the main environmental stresses of drought, strong light, ultraviolet radiation, high temperature, low temperature(day and night or annual temperature difference is large), nutrient deficiency, pests and so on. ②Based on analyzing the strategies of medicinal plants to adapt to environmental stresses, it is pointed out that the synthesis and accumulation of secondary metabolites are the most important strategies of medicinal plants to protect against environmental stresses. In the process of long-term adaptation to specific stress, the accumulation of relevant genetic variation and epigenetic inheritance has become an important condition for the formation of quality of medicinal plants. ③It is proposed that "simulative habitat cultivation" has obvious advantages in balancing growth and secondary metabolism and guaranting the quality of traditional Chinese medicinal materials.

[Pattern and matching technology of ecological planting for Chinese materia medica based on system level].

The ecological agriculture of Chinese materia medica(CMM) has become the most dynamic and promising new field in the global ecological agriculture. The development of ecological planting of CMM has become the national strategy of Chinese traditional medicine agriculture. It has been highly valued and has flourished throughout the country, and has formed some more mature ecological planting models of CMM. Based on the system level, this paper sorts out the common ecological cultivation patterns of CMM, and obtains five basic patterns: landscape pattern at the ecological landscape level, circulation pattern at the ecosystem level, stereo model at the bio-community level, biodiversity patterns at the level of biological populations and well-established models at the level of biological individuals. On this basis, eight common ecological planting techniques of CMM were obtained, includingwild tending techniques, fine agricultural farming techniques, directional cultivation techniques, soil improvement techniques, soil testing and fertilization techniques, mycorrhizal cultivation techniques, green control technology for pests and diseases and facility cultivation techniques.This paper aims to provide theoretical basis for scientific research and popularization and application of CMM ecological planting.

[Pattern of ecological planting for Chinese materia medica based on regional distribution].

As an environment-friendly agriculture, ecological agriculture of Chinese materia medica(CMM) is being implemented in all parts of the country. Due to the stronger dependence on natural environmental conditions, ecological agriculture of CMM shows obvious regional differences in production practice. More mature CMM ecological planting patterns representative of each region were collected. It was found that common types of patterns in various regions of the country mainly included intercropping,intercropping,rotation planting mode, undergrowth planting mode, wild tending planting mode and landscape ecological planting mode. Based on the Construction Plan of National Dao-di Herbs Production Base(2018-2025) and Chinese Medicine Division, this paper systematically sorts out the pattern of ecological planting of CMM in the 8-avenue medicinal materials production areas according to the varieties and regions. The specific pattern of ecological planting of CMM included the ginseng undergrowth planting pattern in northeastern China, the bionics wild ecological planting of the Forsythia suspensa in northern China, the Fritillaria thunbergii-rice rotation in eastern China, the imitation wild planting pattern under the Polygonatum cyrtonema in central China, the planting pattern of the Fructus amomi under forest in southern China, the Ligusticum chuanxiong-rice rotation pattern in the Southwest, wild tending of Glycyrrhiza uralensis in the Northwest, and rhubarb imitation wild planting pattern in Qinghai-Tibet area. Finally, it is expected to provide reference for the screening and popularization of ecological planting patterns of other CMMs in various distribution areas.

[Cultivation strategy on cluster brand of ecological agriculture of Dao-di herbs].

The cluster brand is the embodiment of the core competitiveness of an industry. Developing and cultivating cluster brand of ecological agriculture of Dao-di herbs not only helps to optimize the value chain of the Chinese medicinal materials(CMMs) industry cluster, realize the value-added of the CMMs industry cluster, but also enhance the visibility and influence of the industrial cluster, enhance the core competitiveness of the industrial cluster. This has important practical significance for promoting the "orderly" "safe" and "effective" development of the Dao-di herbs. Based on the industry development status of CMMs, this article introduces several concepts related to cluster brands and their relationships, and focuses on the cultivation models and strategies of cluster brand in the CMMs industry. Based on the current status of the development of the CMMs industry, this article introduces several concepts related to cluster brands and their interrelationships. It discusses the cultivation models and strategies of cluster brands in the CMMs industry, industry associations, Chinese medicine companies and individual growers as the support, insists on the ecological cultivation of authentic medicinal materials and the cultivation of cluster brands. Finally, it points out the direction for the high-quality development of the ecological agriculture of CMMs.

[Core position of secondary metabolism of medicinal plants in ecological planting of Chinese materia medica and its utilization].

This paper summarized the effects of ecological planting on secondary metabolism firstly and pointed out that ecological planting can increase the content of secondary metabolites in plants, especially the content of defensive secondary metabolites. The possible mechanism was analyzed subsequently. Then, we reviewed the induction and utilization of secondary metabolism in the ecological planting of Chinese materia medica from the perspectives of biological control of pests and diseases, promotion of beneficial microorganism accumulation, optimization of mixed planting, regulation of no-tillage and straw cover. In this article, we pointed out that paying close attention to secondary metabolism is the most important feature of ecological planting of Chinese materia medica. Ecological planting can promote the accumulation of secondary metabolites of Chinese materia medica which means can improve the quality of Chinese materia medica, beneficial to the prevention and control of diseases, insects and weeds. Furthermore, lacking of systemic researches,the extensive verifications and systematic in-depth researches on the ecological planting of Chinese materia medica should be carry out urgently.

Functional characterization of three flavonoid glycosyltransferases from Andrographis paniculata.

Andrographis paniculata is an important traditional medicinal herb in South and Southeast Asian countries with diverse pharmacological activities that contains various flavonoids and flavonoid glycosides. Glycosylation can transform aglycones into more stable, biologically active and structurally diverse glycosides. Here, we report three glycosyltransferases from the leaves of A. paniculata (ApUFGTs) that presented wide substrate spectra for flavonoid glycosylation and exhibited multi-site glycosylation on the substrate molecules. They acted on the 7-OH position of the A ring and were able to glycosylate several other different types of compounds. The biochemical properties and phylogenetic analysis of these glycosyltransferases were also investigated. This study provides a basis for further research on the cloning of genes involved in glycosylation from A. paniculata and offers opportunities for enhancing flavonoid glycoside production in heterologous hosts. These enzymes are expected to become effective tools for drug discovery and for the biosynthesis of derivatives via flavonoid glycosylation.

[Research progress in aromatic prenyltransferases originated from microorganisms].

The prenylation of aromatic compounds plays an important role in the natural product research because it not only gives rise to an astounding diversity of primary and secondary metabolites in plants, fungi and bacteria but also enhances the bioactivities and bioavailabilities of these compounds. However, further investigation of prenylated aromatic compounds is frequently hindered due to their low content in nature and difficulties in chemical synthesis. Cloning aromatic prenyltransferase genes followed by heterologous expression would be attractive tools for the chemoenzymatic synthesis of bioactive molecules. This review summarizes the classifications, structural investigations, enzymatic catalysis and other progress in aromatic prenyltransferases originated from microorganisms.

[Chemical constituents from cell suspension cultures of Cudrania tricuspidata].

Ten compounds were isolated and purified from cell suspension cultures of Cudrania tricuspidata with silica-gel column chromatography, semi-preparative HPLC and Sephadex LH-20. On the base of their physicochemical properties and spectral data, their structures were identified as 1, 3, 5-trihydroxy4-(3, 3-dimethylallyl) xanthone (1), wighteone (2), 6-prenylapigenin (3), licoflavone C(4), cudraflavanone C(5), erythrivarone A (6), derrone (7), carthamidin (8), genistein (9) and aromadendrin (10). Among them, compounds 2-10 were flavonoids, and compound 1 was a xanthone which was isolated from the plant for the first time.

[Changes of dietary pattern among adults in Liaoning province, 1989 to 2006].

OBJECTIVE: To study the changes of dietary pattern among adult residents in different areas of Liaoning province from 1989 to 2006. METHODS: Healthy adults (6213 subjects) at age of 18 - 65 years from 480 households in three cities (Shenyang, Yingkou, Wafangdian) and three counties (Qingyuan, Huanren, Chaoyang) were selected with stratified multiple cluster random sampling. The information on nutrient intake of the subjects were collected from datasets of Chinese Health and Nutrition Survey conducted in 1989, 1991, 1993, 2000, 2004, and 2006. Different food intake, the nutrients intake percentages for recommended nutrition intake (RNI) and appropriate intake (AI), and the percentages of total energy and protein from grain, animal product, bean and its product were calculated to assess the residents' dietary pattern and nutrition status. The changes of dietary pattern among adult residents were analyzed. RESULTS: Among the residents, there were a 38.1% of decreased intake for grain (from 601.9 to 372.5 g/d), 20.5% for potato (from 75.6 to 60.1 g/d), 25.1% for beans (from 38.7 to 29.0 g/d), and a 77.2% of increased intake for fish and shrimp (from 25.0 to 44.3 g/d), 36.9% for livestock and poultry (from 65.6 to 89.8 g/d), 47.7% for fruit (from 70.7 to 104.4 g/d), and intake of milk product (from 5.8 to 21.3 g/d), egg (from 17.3 to 35.7 g/d), vegetable (from 296.1 to 316.3 g/d) were also increased from 1989 to 2006. During the period, the intake percentages of energy and protein from grain decreased from 67.5% (8.7 MJ/12.8 MJ per day) to 51.5% (5.0 MJ/9.6 MJ per day) and from 72.0% (66.2 g/91.9 g per day) to 59.7% (45.3 g/75.9 g per day), and on the contrary those from animal products increased from 8.9% (1.1 MJ/12.8 MJ per day) to 14.8 (1.4 MJ/9.6 MJ per day) and from 15.9% (14.6 g/91.9 g per day) to 27.9% (21.2 g/75.9 g per day), respectively. In 2006, the intake of vitamin A (508.9 µg/d) was 67.6% of it's RNI, intake of vitamin B(2) (0.9 mg/d) was 64.6% and the intake of calcium (453.7 mg/d) was 52.5% of it's AI among the residents. CONCLUSION: The intake of plant food decreased and that of animal food increased from 1989 to 2006 and the dietary intakes of calcium, vitamin A, vitamin B(2) need to be increased among adult population of Liaoning province.

[Identification and clone of human Alzheimer's disease related gene nicastrin promoter].

OBJECTIVE: To identify the promoter of human nicastrin (NCT) gene, a major component of gamma-secretase which is closely related with pathogenesis of Alzheimer's disease. METHODS: Promoter of human Alzheimer's disease related gene, nicastrin, a 1768 bp fragment was firstly isolated from human genomic DNA by PCR. This fragment's 3 flanking end was 4 bp upstream to the start codon ATG (+1) of the gene. This fragment was used as template, a series of deleted fragments were amplified and constructed to the pGL3-Enhancer plasmid with the artificial designed linkers. The relative activity of their promoter in Hela cells was studied by dual-luciferase assay. RESULTS: The 420 bp fragment showed the strongest activity, and the 237 bp fragment was the minimal fragment in length with activity. CONCLUSION: The promoter of NCT is located at -432/-133 region upstream the translational start codon, while its basal promoter is between -359/-90 that drives the transcription of reporter gene in Hela cells.