Transcriptome Reveals Long Non-coding RNAs and mRNAs Involved in Primary Wool Follicle Induction in Carpet Sheep Fetal Skin.

Murine primary hair follicle induction is driven by the communication between the mesenchyme and epithelium and mostly governed by signaling pathways including wingless-related integration site (WNT), ectodysplasin A receptor (EDAR), bone morphogenetic protein (BMP), and fibroblast growth factor (FGF), as observed in genetically modified mouse models. Sheep skin may serve as a valuable system for hair research owing to the co-existence of sweat glands with wool follicles in trunk skin and asynchronized wool follicle growth pattern similar to that of human head hair follicles. However, the mechanisms underlying wool follicle development remain largely unknown. To understand how long non-coding RNAs (lncRNAs) and mRNAs function in primary wool follicle induction in carpet wool sheep, we conducted high-throughput RNA sequencing and revealed globally altered lncRNAs (36 upregulated and 26 downregulated), mRNAs (228 elevated and 225 decreased), and 80 differentially expressed novel transcripts. Several key signals in WNT (WNT2B and WNT16), BMP (BMP3, BMP4, and BMP7), EDAR (EDAR and EDARADD), and FGF (FGFR2 and FGF20) pathways, and a series of lncRNAs, including XLOC_539599, XLOC_556463, XLOC_015081, XLOC_1285606, XLOC_297809, and XLOC_764219, were shown to be potentially important for primary wool follicle induction. GO and KEGG analyses of differentially expressed mRNAs and potential targets of altered lncRNAs were both significantly enriched in morphogenesis biological processes and transforming growth factor-ß, Hedgehog, and PI3K-Akt signaling, as well as focal adhesion and extracellular matrix-receptor interactions. The prediction of mRNA-mRNA and lncRNA-mRNA interaction networks further revealed transcripts potentially involved in primary wool follicle induction. The expression patterns of mRNAs and lncRNAs of interest were validated by qRT-PCR. The localization of XLOC_297809 and XLOC_764219 both in placodes and dermal condensations was detected by in situ hybridization, indicating important roles of lncRNAs in primary wool follicle induction and skin development. This is the first report elucidating the gene network of lncRNAs and mRNAs associated with primary wool follicle early development in carpet wool sheep and will shed new light on selective wool sheep breeding.

Defining Key Genes Regulating Morphogenesis of Apocrine Sweat Gland in Sheepskin.

The apocrine sweat gland is a unique skin appendage in humans compared to mouse and chicken models. The absence of apocrine sweat glands in chicken and murine skin largely restrains further understanding of the complexity of human skin biology and skin diseases, like hircismus. Sheep may serve as an additional system for skin appendage investigation owing to the distributions and histological similarities between the apocrine sweat glands of sheep trunk skin and human armpit skin. To understand the molecular mechanisms underlying morphogenesis of apocrine sweat glands in sheepskin, transcriptome analyses were conducted to reveal 1631 differentially expressed genes that were mainly enriched in three functional groups (cellular component, molecular function and biological process), particularly in gland, epithelial, hair follicle and skin development. There were 7 Gene Ontology (GO) terms enriched in epithelial cell migration and morphogenesis of branching epithelium that were potentially correlated with the wool follicle peg elongation. An additional 5 GO terms were enriched in gland morphogenesis (20 genes), gland development (42 genes), salivary gland morphogenesis and development (8 genes), branching involved in salivary gland morphogenesis (6 genes) and mammary gland epithelial cell differentiation (4 genes). The enriched gland-related genes and two Kyoto Encyclopedia of Genes and Genomes pathway genes (WNT and TGF-ß) were potentially involved in the induction of apocrine sweat glands. Genes named BMPR1A, BMP7, SMAD4, TGFB3, WIF1, and WNT10B were selected to validate transcript expression by qRT-PCR. Immunohistochemistry was performed to localize markers for hair follicle (SOX2), skin fibroblast (PDGFRB), stem cells (SOX9) and BMP signaling (SMAD5) in sheepskin. SOX2 and PDGFRB were absent in apocrine sweat glands. SOX9 and SMAD5 were both observed in precursor cells of apocrine sweat glands and later in gland ducts. These results combined with the upregulation of BMP signaling genes indicate that apocrine sweat glands were originated from outer root sheath of primary wool follicle and positively regulated by BMP signaling. This report established the primary network regulating early development of apocrine sweat glands in sheepskin and will facilitate the further understanding of histology and pathology of apocrine sweat glands in human and companion animal skin.

Glucose intolerance associated with hypoxia in people living at high altitudes in the Tibetan highland.

OBJECTIVES: To clarify the association between glucose intolerance and high altitudes (2900-4800 m) in a hypoxic environment in Tibetan highlanders and to verify the hypothesis that high altitude dwelling increases vulnerability to diabetes mellitus (DM) accelerated by lifestyle change or ageing. DESIGN: Cross-sectional epidemiological study on Tibetan highlanders. PARTICIPANTS: We enrolled 1258 participants aged 40-87 years. The rural population comprised farmers in Domkhar (altitude 2900-3800 m) and nomads in Haiyan (3000-3100 m), Ryuho (4400 m) and Changthang (4300-4800 m). Urban area participants were from Leh (3300 m) and Jiegu (3700 m). MAIN OUTCOME MEASURE: Participants were classified into six glucose tolerance-based groups: DM, intermediate hyperglycaemia (IHG), normoglycaemia (NG), fasting DM, fasting IHG and fasting NG. Prevalence of glucose intolerance was compared in farmers, nomads and urban dwellers. Effects of dwelling at high altitude or hypoxia on glucose intolerance were analysed with the confounding factors of age, sex, obesity, lipids, haemoglobin, hypertension and lifestyle, using multiple logistic regression. RESULTS: The prevalence of DM (fasting DM)/IHG (fasting IHG) was 8.9% (6.5%)/25.1% (12.7%), respectively, in all participants. This prevalence was higher in urban dwellers (9.5% (7.1%)/28.5% (11.7%)) and in farmers (8.5% (6.1%)/28.5% (18.3%)) compared with nomads (8.2% (5.7%)/15.7% (9.7%)) (p=0.0140/0.0001). Dwelling at high altitude was significantly associated with fasting IHG+fasting DM/fasting DM (ORs for >4500 and 3500-4499 m were 3.59/4.36 and 2.07/1.76 vs <3500 m, respectively). After adjusting for lifestyle change, hypoxaemia and polycythaemia were closely associated with glucose intolerance. CONCLUSIONS: Socioeconomic factors, hypoxaemia and the effects of altitudes >3500 m play a major role in the high prevalence of glucose intolerance in highlanders. Tibetan highlanders may be vulnerable to glucose intolerance, with polycythaemia as a sign of poor hypoxic adaptation, accelerated by lifestyle change and ageing.

[Echinococcosis prevalence in elderly population from Haiyan County of Qinghai Province].

Echinococcus infection in elderly population (60-87 years old) was investigated by serological assay, medical imaging and epidemiology history in August, 2008. A total of 234 subjects from different nationalities were examined. Seven cases were serologically positive for echinococcosis and confirmed by medical imaging. Out of 30 suspected sera-positive cases, 3 were positive by medical imaging. There were 10 confirmed echinococcosis cases with a positive rate of 4.3%. There was significant difference on the prevalence among different towns (P < 0.05), but no significant statistical difference on the prevalence among nationalities, careers and sex. The prevalence in females (4.8%) was higher than that of males (3.6%).

Comprehensive geriatric assessment of elderly highlanders in Qinghai, China I: activities of daily living, quality of life and metabolic syndrome.

AIM: To reveal the comparison of comprehensive geriatric functions of elderly highlanders in Qinghai Plateau in China among three different ethnic groups. METHODS: Activities of daily living (ADL), screening-based depression, quality of life (QOL) and checking-up of metabolic syndrome including community-based oral glucose tolerance test were assessed in 393 community-dwelling elderly subjects aged 60 years or more (247 Han elderly subjects, 49 Mongolian ones and 97 Tibetan ones). RESULTS: Tibetan elderly highlanders were more disabled in ADL, but had higher QOL than Han elderly ones in Qinghai Plateau. Blood pressure measurements, rate of hypertension and hemoglobin concentrations in Tibetan elderly highlanders were lower than Han ones. Rates of diabetes and impaired glucose tolerance in elderly highlanders were relatively lower than other Asian elderly lowlanders. CONCLUSION: Prevalence of metabolic syndrome in elderly highlanders in Qinghai was still not high, however, we should pay attention to its tendency related with socialglobalism in the near future. Further investigation on physiological adaptability to hypoxic environment and human ageing phenomena in a global context may open a new research frontier for ageing science.

Comprehensive geriatric assessment of elderly highlanders in Qinghai, China II: the association of polycythemia with lifestyle-related diseases among the three ethnicities.

AIM: The objective of this study is to disclose the association of polycythemia with lifestyle-related diseases (hypertension, obesity and glucose intolerance) among the three ethnicities in Qinghai, China. METHODS: The subjects were 393 elderly people (247 Han, 97 Tibetan and 49 Mongolian) aged 60 years and more living in Qinghai (3000 m a.s.l.) in China. The associated factors with polycythemia were analyzed in the subjects. Excessive polycythemia was defined as hemoglobin concentration over 20 mg/dL. RESULTS: Polycythemia was associated with men, hypoxemia, obesity and high diastolic blood pressure (DBP) in the elderly in Qinghai. Male sex was associated with polycythemia in all ethnicities. Obesity was associated with Han and Tibetan men. Glucose intolerance and activities of daily living were not directly associated with polycythemia after adjustment for sex. There were 7.9% with excessive polycythemia. Independently-associated factors for excessive polycythemia were male sex, body mass index of 25 or more, SpO(2) of less than 85%, DBP of 85 mmHg or more and Han ethnicity (vs Tibetan) by multiple logistic regression. CONCLUSION: There was a close association of polycythemia with diastolic hypertension and obesity in lifestyle-related diseases in high-altitude elderly people. Han people had a higher hemoglobin concentration after adjustment of lifestyle-related diseases compared with Tibetan people. The difference of hemoglobin concentration may be due to Tibetans undergoing a much longer period of adaptation than Han people. Further study is needed to disclose the association between the difference of hypoxic adaptation, lifestyle-related diseases and chronic mountain sickness for their prevention.

Comprehensive geriatric assessment of elderly highlanders in Qinghai, China, III: oxidative stress and aging in Tibetan and Han elderly highlanders.

BACKGROUND: Although there are several factors which may contribute to oxidative stress at high altitude, little is known about the association between oxidative stress and aging in the community-dwelling elderly in the Tibetan Plateau. METHODS: Reactive oxygen species (ROS) and comprehensive geriatric functions were examined among 235 community-dwelling elderly subjects aged 60 years or more (146 Hans and 89 Tibetans). As a marker of ROS, the levels of reactive oxygen metabolites (ROM) were measured using the d-ROM test. RESULTS: The rate of dependence of basic activities of daily living (basic ADL) among Tibetan elderly highlanders was significantly higher than that among Han elderly highlanders. The d-ROM level was higher among the Tibetan elderly than those among the Han elderly (Tibetan 465.6 +/- 97.9 Carr U, Han 415.3 +/- 72.0 Carr U, P = 0.003). The ROM level was higher among women than those among men. Stepwise multiple regression analysis showed that being Tibetan, female, and oxygen saturation were independent predictors of increasing d-ROM level (Tibetan beta, 0.241; female beta, 0.206; oxygen saturation beta, 0.218). The high levels of ROM (d-ROM >500 Carr U) were significantly associated with dependence of basic ADL after adjustment for age, sex and ethnicity (odds ratio = 2.51, P = 0.028). CONCLUSION: The findings of this study imply the possibility that ROS is higher among Tibetan elderly highlanders than that of Han, which related to the geriatric items. Further studies are needed to show the impact of oxidative stress on the aging of highlanders.

Comprehensive geriatric assessment of elderly highlanders in Qinghai, China IV: comparison of food diversity and its relation to health of Han and Tibetan elderly.

AIM: To examine the association between food diversity and health status of Han and Tibetan elderly highlanders in Qinghai Plateau, China. METHODS: The study population consisted of 240 community-dwelling elderly subjects aged 60 years or more (176 Han elderly subjects, 64 Tibetan ones). Food diversity was determined using an 11-item Food Diversity Score Kyoto (FDSK-11). Subjects were interviewed on health status including activities of daily living (ADL), screening-based depression and quality of life (QOL). Blood chemical investigation was carried out in association with food diversity. RESULTS: ADL was significantly lower in both Han and Tibetan elderly with lower food diversity than those with higher diversity. In Han elderly with lower food diversity, QOL was significantly lower in the items of subjective sense of health, relationship with family and subjective happiness, but not significant in Tibetan elderly. A close association was found between lower food diversity and lower financial satisfaction in both Han and Tibetan subjects. No association was found between food diversity and age or body mass index. Higher food diversity was associated with lower blood glucose level in Han elderly subjects, but the opposite association was found in Tibetan ones. CONCLUSION: Food diversity was associated with ADL and QOL in highlanders in Qinghai, China. Food assessment is very important as a useful indicator to establish the actual condition of diet and its relation to health status of community-dwelling elderly as well as the change of economic background in the Qinghai highlands.