Long-term Care Facility and its Elderly Chronic Diseases in Jishou: Insights into Underdeveloped Area of China.

Insight into the current status of long-term care facilities (LTCFs) and chronic diseases in underdeveloped areas in China is scant. Using a census method to survey older adults ≥60 years old (154 older adult residents) in all LTCFs in Jishou area. The International Resident Assessment Instrument for Long-Term Care Facilities (interRAI-LTCF) was used to collect information and analyze the current status of chronic diseases among older adult residents. There were 62 187 residents ≥60 years old in Jishou area. According to the survey, there were only 154 older adult residents living in the LTCFs of Jishou, with a ratio was 0.3% of all older adult residents, which was much lower than China's 3.0%. Of respondents (109 older adult residents), the prevalence of chronic diseases was 70.6%. The prevalence increases with age, as well as in the female is higher than in the male. The top 3 chronic diseases were hypertension (41.3%), bone and joint disease (12.8%) and cerebrovascular disease (12.8%). It was found that different age groups and genders lead to differences in the prevalence and the order of chronic diseases. The prevalence of older adults with impaired balance ability, sleep disturbance and swallowing disturbance was higher than that of normal older adults. The results of the one-way analysis of variance showed that the age and balance ability of the older adults with chronic diseases were statistically significant (P < .05). In addition, the prevalence of chronic diseases in the LTCFs older adult was higher than the home care (HC) older adults in Jishou. The age and the prevalence of chronic diseases of LTCFs in older adults with professional nursing staff were higher than in those without. This study provides a theoretical basis for the healthcare distribution, prevention and treatment of chronic diseases in underdeveloped areas. The undeveloped area has lower LTCFs staying rate and chronic disease prevalence of older adults than relatively developed areas. More public health attention and capital investment are needed to increase the LTCFs number and strengthen disease testing and screening. As well as, the proportion of professional nursing staff and specialist doctors in LTCFs and improve the quality of care and medical treatment for older adults.

Tabrizicola alkalilacus sp. nov., isolated from alkaline Lake Dajiaco on the Tibetan Plateau.

A novel Gram-stain-negative, aerobic, non-motile and rod-shaped bacterium was isolated from Lake Dajiaco on the Tibetan Plateau. Strain DJCT grew without NaCl and tolerated up to 3 % (w/v) NaCl. Growth occurred at pH 6.0-10.0 (optimum, pH 7.0-8.0) and 15-37 °C (optimum, 25-30 °C). Vitamins were not required for growth. The main polar lipids of strain DJCT were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The predominant respiratory quinone was Q-10. The major fatty acid was C18 : 1ω7c. Genome sequencing revealed a genome size of 4.61 Mbp and a G+C content of 62.9 mol%. Analysis of 16S rRNA sequences showed that strain DJCT belonged to the genus Tabrizicola, with the closest neighbour Tabrizicola aquatica RCRI19T (97.5 %). DNA-DNA relatedness between strain DJCT and the closest phylogenetically related strain T. aquatica RCRI19T was 40.8 %. Stain DJCT was clearly distinguished from the type strain mentioned above through phylogenetic analysis, DNA-DNA hybridization, fatty acid composition data and a range of physiological and biochemical characteristic comparisons. Based on its phenotypic and chemotaxonomic characteristics, strain DJCT could be classified as a representative of a novel species of the genus for which the name Tabrizicola alkalilacus sp. nov. is proposed. The type strain is DJCT (=CICC 24242T=KCTC 62173T).

Optimization of Ultrasound-Assisted Extraction, HPLC and UHPLC-ESI-Q-TOF-MS/MS Analysis of Main Macamides and Macaenes from Maca (Cultivars of Lepidium meyenii Walp).

Ultrasound-assisted extraction (UAE), using petroleum ether as the solvent, was systematically applied to extract main macamides and macaenes from Maca hypocotyls. Extraction yield was related with four variables, including ratio of solution to solid, extraction temperature, extraction time, and extraction power. On the basis of response surface methodology (RSM), the optimal conditions were determined to be the ratio of solution to solid as 10:1 (mL/g), the extraction temperature of 40 °C, the extraction time of 30 min, and the extraction power of 200 W. Based on the optimal extraction method of UAE, the total contents of ten main macamides and two main macaenes of Maca cultivated in twenty different areas of Tibet were analyzed by HPLC and UHPLC-ESI-Q-TOF-MS/MS. This study indicated that UAE was able to effectively extract macamides alkaloids from Maca hypocotyls. Quantitative analysis showed that geographical origins, not ecotypes, played a more important role on the accumulation of active macamides in Maca.

New Germplasms of the Culinary-Medicinal Button Mushroom, Agaricus bisporus (Agaricomycetes): Two Wild Strains from the Tibetan Plateau (China).

Agaricus bisporus is one of the most important commercially cultivated culinary-medicinal mushrooms worldwide. In China, most of the cultivated strains of the fungus were introduced from other countries and cultivated in the eastern provinces. In this study, 2 wild strains of A. bisporus, 2091 and 2094, isolated from fresh specimens collected from the Tibetan Plateau, were domesticated and cultivated alongside a commercial hybrid strain, As2796, in Lhasa, China, for comparison in order to provide new germplasms for cultivation. Basic characteristics, mushroom yield, dry weight, polysaccharide contents, and antioxidant activities of the tested strains were analyzed. Compared with strain As2796, the 2 wild strains displayed good values for mycelial growth, time to fruiting, mushroom yield, dry weight, and polysaccharide contents, and their basidiomata had distinct morphological characteristics (e.g., brown or pale brown caps with some white scales). In addition, the antioxidant activities (reducing power and DPPH radical scavenging effect) of strain 2094 were significantly higher than those of the other 2 strains. Domestication of the 2 wild strains would add more genetic variation into the germplasm of A. bisporus for cultivation, especially in China, and might help to address the problem inherent to the nearly monoculture crop lacking genetic diversity in China.

Bacterial diversity in native habitats of the medicinal fungus Ophiocordyceps sinensis on Tibetan Plateau as determined using Illumina sequencing data.

Ophiocordyceps sinensis is one of the most well-known traditional Chinese medicinal fungi. In this study, bacterial diversity in the soils of native habitats of O. sinensis was investigated using Illumina sequencing data. A total of 525,000 sequences of V6-16S rRNA were analyzed. The number of OTUs from each sample ranged from 13,858 to 15,978 at 97% sequence similarity cut-off. The results demonstrated that the deep sequencing approach provides improved access to rare genotypes. Richness indices and Shannon's diversity index did not differ significantly between samples collected from locations where O. sinensis was present (Os1-3) and not present (NOs1-3). Classified bacterial sequences were grouped into 23 phyla including Proteobacteria, Actinobacteria, Acidobacteria, Verrucomicrobia, etc. The Venn diagram revealed that 7183 OTUs belonging to 14 phyla were shared by Os, NOs and MP (mycelial pellicle wrapping the sclerotium of O. sinensis) samples, possibly representing a core microbiome existing in native habitats of O. sinensis, and that 863 belonging to 12 phyla were shared by Os and MP samples, possibly related to the occurrence of O. sinensis. Overall, the results revealed a high bacterial diversity in the soil samples and the relationships between the bacterial diversity and O. sinensis merit further investigation.

Two new macrosporin dimers from the fungus Alternaria sp. XZSBG-1.

Two new macrosporin dimers (1-2) along with four known compounds (3-6) were isolated from the extracts of the fungal strain Alternaria sp. XZSBG-1 from the sediment of the salt lake in the Bange, Tibetan, China. Their structures were determined by spectroscopic methods, mainly by 2D NMR spectra. Compounds 1 and 2 are new macrosporin dimers with symmetric chemical structures. In the cytotoxicity assay and inhibited alpha-glucosidase activity assay, all these compounds showed no notable inhibitory activity.

Extracts from the roots of Incarvillea younghusbandii on antioxidant effects and life span prolonging in Drosophila melanogaster.

AIM: To investigate antioxidant activities and life span prolonging effects of the extracts from the roots of Incarvillea younghusbandii Sprague, and to study the correlations between these activities and the polar intensity of the extracts. METHOD: Five extracts (IYS1, IYS2, IYS3, IYS4 and YS5) with different polar intensity were prepared. Antioxidant activities in vitro were determined by LPO inhibitory and free radicals scavenging experiments. Life span prolonging effects in vivo were evaluated by feeding Drosophila melanogaster. RESULT: Total phenolic content in extracts were solvent-dependent and decreased in the order of IYS4 > IYS1 >> IYS3 > IYS5 > IYS2. Organic extracts (IYS1 and IYS4) showed excellent LPO inhibitory activity, O(2)(· -) and ·OH scavenging activity compared to ascorbic acid (or benzoic acid, or BHT), while aqueous extracts (IYS2, IYS3 and IYS5) did not. The antioxidant activities (in vitro) were solvent dependent and decreased in the order of IYS4 > IYS1 > IYS3 > IYS5 ≥ IYS2. Drosophila melanogaster was fed with organic extracts (IYS1 or IYS4) at 5.0 mg mL(-1). The mean life span were increased by 24.4% (IYS1) or 23.0% (IYS4) in female and 15.3% (IYS1) or 16.9% (IYS4) in male; the maximum life span were increased by 8.4% (IYS1) or 11.2% (IYS4) in female and 9.7% (IYS1) or 15.8% (IYS4) in male, and the survival curves were significantly shifted to the right after fifteen days in both sexes survival period. Feeding aqueous extracts (IYS2, IYS3 or IYS5) at 5.0 mg·mL(-1), the significant life span prolonging effects were not achieved. The life span prolonging effects of the extracts were solvent-dependent and decreased in the order of IYS4 ≥ IYS1 >> IYS3 > IYS2 > IYS5. CONCLUSION: Extracts from the roots of Incarvillea younghusbandii Sprague showed excellent antioxidant activities and significant life span prolonging effects in Drosophila melanogaster. Positive correlations existed between the antioxidant activities and total phenolic content. Life span prolonging effect was positively correlated with the total phenolic content or antioxidant activities. The extracts possess better life span prolonging effect in females than in males.

A survey of the geographic distribution of Ophiocordyceps sinensis.

Ophiocordyceps sinensis is one of the best known fungi in Traditional Chinese Medicine. Many efforts have been devoted to locating the production areas of this species resulting in various reports; however, its geographic distribution remains incompletely understood. Distribution of O. sinensis at the county level is clarified in this work based on both a literature search and fieldwork. More than 3600 publications related to O. sinensis were investigated, including scientific papers, books, and online information. Herbarium specimens of O. sinensis and field collections made by this research group during the years 2000-2010 were examined to verify the distribution sites. A total of 203 localities for O. sinensis have been found, of which 106 are considered as confirmed distribution sites, 65 as possible distribution sites, 29 as excluded distribution sites and three as suspicious distribution sites. The results show that O. sinensis is confined to the Tibetan Plateau and its surrounding regions, including Tibet, Gansu, Qinghai, Sichuan, and Yunnan provinces in China and in certain areas of the southern flank of the Himalayas, in the countries of Bhutan, India and Nepal, with 3,000 m as the lowest altitude for the distribution. The fungus is distributed from the southernmost site in Yulong Naxi Autonomous County in northwestern Yunnan Province to the northernmost site in the Qilian Mountains in Qilian County, Qinghai Province, and from the east edge of the Tibetan Plateau in Wudu County, Gansu Province to the westernmost site in Uttarakhand, India. The clarification of the geographic distribution of O. sinensis will lay the foundation for conservation and sustainable use of the species.

Isolation, purification and structure identification of two phenolic glycosides from the roots of Incarvillea younghusbandii Sprague and their antioxidant activities.

Using a bioassay-guided fractionation technique, two compounds were isolated from the roots of Incarvillea younghusbandii Sprague through silica gel, reverse-phase C18 column chromatography and reverse-phase HPLC. Their structures were identified as acteoside (1) and isoacteoside (2) by ESI-MS, GC-MS, 1D- and 2D-NMR. 1 and 2 showed *OH scavenging capacity similar with benzoic acid, higher O2*- (or *OH) scavenging capacity than ascorbic acid, far higher hepatic LPO inhibitory activities than 2, 6-di-tert-butyl-4-methylphenol (BHT) or ascorbic acid, and more powerful effect on protecting erythrocytes from oxidative damage than ascorbic acid. The *OH scavenging capacity was positively proportional to the concentrations of 1 and 2 ranging from 0.015 6 to 0.500 0 mg x mL(-1). The hepatic LPO inhibitory activities increased with the increasing concentrations of 1 and 2 from 0.001 9 to 0.250 0 mg x mL(-1), but decreased slightly with the increasing concentration from 0.250 0 to 1.0000 mg x L(-1).

Composition analysis and anti-hypoxia activity of polysaccharide from Brassica rapa L.

Crude water-soluble polysaccharides (BRP) were extracted from the root of Brassica rapa L. using boiling-water. The polysaccharides were successively purified by chromatography on DEAE-cellulose and Sephadex G-100 column, giving three major polysaccharide fractions termed BRP1-1, BRP2-1, BRP3-1. The gel permeation chromatography (GPC) analysis showed that the average molecular weight (Mw) of polysaccharides (BRP1-1, BRP2-1, BRP3-1) were approximately 5.53×10(3) Da, 3.35×10(4) Da and 3.37×10(4) Da, respectively. Monosaccharide components analysis indicated that BRP1-1 was composed of arabinose and glucose in a molar ratio of 1.66:98.34. BRP2-1 was composed of arabinose, galactose and glucose in a molar ratio of 9.3:14.63:76.07. BRP3-1 was composed of arabinose, rhamnose, galactose and glucose in a molar ratio of 24.98:24.10:44.09:6.83. The evaluation of anti-hypoxia activity in vivo revealed that BRP is a novel potential anti-hypoxia agent.

[Bacterial diversity of disused thermal vents in Tibetan Yangbajing Region].

OBJECTIVE: To study the bacterial diversity in sediment and water from two disused thermal vents in Yangbajing, Tibet, China. METHODS: We constructed 16S rRNA gene libraries of the total DNA from three samples. Sediment sample A and water sample A were from thermal vent A. Sediment sample B was from thermal vent B. Positive clones from the libraries were analyzed randomly by amplified ribosomal DNA restriction analysis. Positive clones' sequences of every Operational Taxonomic Units from libraries were determined, and sequence data were submitted to GenBank and contrasted to those known sequences. Phylogenetic trees were built up by using MEGA4.0 program. RESULTS: Most of bacteria communities of the two thermal vents were typical thermophilc inhabitants, the predominant proteobacteria communities were found in thermal vent A and thermal vent B (ratios in sediment A, water sample A and sediment B were 41.08%, 38.00% and 42.57%) One of the sub-predominant bacteria communities, Deinococcus-Thermus, was found in thermal vent A and thermal vent B (ratios in sediment A, water sample A and sediment B were 10.71%, 20.00% and 21.30%). Moreover, one of the sub-predominant bacteria communities, Acidobacteria that was rarely found in hot springs or thermal vents, was present both in sediments of thermal vent A and B (16.07%, 19.15%). The third sub-predominant community in sediment of thermal vent A was Eubacterium sp. (14.28%), belonging to phylum Firmicutes whereas no Acidobacteria was found in water sample A. Instead, Hydrogenobacter, belonging to phylum Aquificae, was another sub-predominant community (16.00%) in water sample A. In addition, Chloroflexi, Cyanobacteria and CFB group ( Cytophaga-Flexibacter-Bacteroides) were detected in both thermal vents. CONCLUSION: Compared with and contrasted to the references, we found that bacteria communities in Yangbajing thermal field were similar to that in some hot springs and thermal vents around the world but slightly abnormal. Some bacteria communities in thermal vent A and thermal vent B were not very popular, such as Vibrio sp., Bacteriovorax sp., HolophagalAcidobacterium, Verrucomicrobium.

Isolation, purification and structure identification of antioxidant compound from the roots of Incarvillea younghusbandii Sprague and its life span prolonging effect in Drosophila melanogaster.

For the purpose of preventative treatment for oxidative stress-mediated diseases and anti-aging, a high-antioxidant compound was isolated from the roots of Incarvillea younghusbandii Sprague for the first time through silica gel column chromatography, reverse-phase C18 column chromatography and reverse-phase semi-preparative HPLC using a bioassay-guided fractionation technique, and was identified as acteoside by ESI-MS, GC-MS, 1D- and 2D-NMR. Feeding Drosophila melanogaster with acteoside, significant life span prolonging effect were achieved at the dosage range from 0.64 to 2.56 mg mL(-1). Positive relationships existed between the dosage and the life span prolonging effect. Otherwise, acteoside possess better life span prolonging effect in females than in males.

[Experimental study on the antiviral mechanism of Ceratostigma willmattianum against herpes simplex virus type 1 in vitro].

OBJECTIVE: To study the antiviral effect and mechanisms of the liquid extract from Ceratostigma willmattianum against herpes simplex virus type 1 (HSV-1) in vitro. METHOD: C. willmattianum in various concentration was applied to different steps of HSV-1 replication cycle. 50% Tissue culture infective dose (TCID50), cytopathic effect (CPE), MTT staining method, dot blotting and Northern blotting analysis were used to estimate index of antiviral activity. RESULT: 50% Toxic concentration (TC50) was 1077 mg x L(-1), IC50 29.46 mg x L(-1) and therapeutic index (TI) 36.56 in C. willmattianum. TC50 330 mg x L(-1), 50% Inhibiting concentration (IC50) 9.12 mg x L(-1) and TI 36.18 in ACV by MTT staining method. The liquid extract from C. willmattianum had remarkable effect on inhibiting HSV-1 in vitro. Ceratostigma could interfere absorption of HSV-1 to Vero cells to prevent HSV-1 infectivity, inhibit HSV-1 gD DNA replication and HSV-1 gD mRNA expression. CONCLUSION: C. willmattianum possesses strong anti-HSV-1 activity in vitro. The antiviral mechanisms are related to inhibiting virus absorption, HSV-1 gD gene replication and HSV-1 gD gene transcription.