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The objective of this study was to assess the genetic diversity, phylogenetic relationship and population structure of five goat breeds in Shanxi, China. High genetic diversities were found in the five populations, among which, Licheng big green goat (LCBG) has the highest genetic diversity, while Jinlan cashmere goat (JLCG) population has the lowest genetic diversity. Bottleneck analysis showed the absence of recent genetic bottlenecks in the five goat populations. Genetic differentiation analysis shows that the closest genetic relationship between LCBG and LLBG (Lvliang black goat) was found, and the genetic distance between JLCG and the other four populations is the largest. The population structure of JLCG is different from the other four populations with K = 2, while LCBG and LLBG have high similarity population structure as the K value changes. Knowledge about genetic diversity and population structure of indigenous goats is essential for genetic improvement, understanding of environmental adaptation as well as utilization and conservation of goat breeds.
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Variação Genética , Genética Populacional , Animais , Variação Genética/genética , Filogenia , Cabras/genética , Repetições de Microssatélites/genéticaRESUMO
In this paper, a monolayer SiO2 microsphere (MS) array was self-assembled on a silicon substrate, and monolayer dense silver nanoparticles (AgNPs) with different particle sizes were transferred onto the single-layer SiO2 MS array using a liquid-liquid interface method. A double monolayer "Ag@SiO2" with high sensitivity and high uniformity was prepared as a surface-enhanced Raman scattering (SERS) substrate. The electromagnetic distribution on the Ag@SiO2 substrate was analyzed using the Lumerical FDTD (finite difference time domain) Solutions software and the corresponding theoretical enhancement factors were calculated. The experimental results show that a Ag@SiO2 sample with a AgNPs diameter of 30 nm has the maximal electric field value at the AgNPs gap. The limit of detection (LOD) is 10-16 mol/L for Rhodamine 6G (R6G) analytes and the analytical enhancement factor (AEF) can reach ~2.3 × 1013. Our sample also shows high uniformity, with the calculated relative standard deviation (RSD) of ~5.78%.
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Nanopartículas Metálicas , Prata , Nanopartículas Metálicas/química , Microesferas , Dióxido de Silício/química , Prata/química , Análise Espectral Raman/métodosRESUMO
BACKGROUND The aim of this study was to investigate the effect of ribosomal protein L22 (RPL22) on gastric cancer (GC) cell proliferation, migration, and apoptosis, and its correlation with the murine double minute 2-protein 53 (MDM2-p53) signaling pathway. MATERIAL AND METHODS The RPL22 expression in GC tissues and cells was detected by quantitative reverse transcription-polymerase chain reaction and western blotting. RPL22 was overexpressed in the MKN-45 cells by the transfection of a vector, pcDNA3.1 (pcDNA)-RPL22, whereas it was silenced in the MGC-803 cells by the transfection of short interfering (si) RNA (si-RPL22). Flow cytometric analysis, cell viability assays, wound healing assays, and transwell assays were utilized to explore the influences of RPL22 on the apoptosis, proliferation, migration, and invasion. Nutlin-3 (an MDM2-p53 inhibitor) was used to inhibit MDM2-p53 signaling. RESULTS The RPL22 expression was downregulated in GC tissues and cells. It was significantly lower in the advanced GC tissues than in the early GC tissues, and was significantly lower in the lymphatic metastatic tissues than in the non-lymphatic metastatic tissues. The transfection of si-RPL22 accelerated the ability of GC cells to proliferate and metastasize, whereas apoptosis was dampened. The transfection of pcDNA-RPL22 exerted the opposite effect on the GC cells; MDM2 expression was upregulated in RPL22-silenced GC cells, while the expression of p53 was downregulated. In vitro, treatment with nutlin-3 reversed the promoting effects of si-RPL22 on GC progression. CONCLUSIONS In vitro, the silencing of RPL22 aggravates GC by regulating the MDM2-p53 signaling pathway.
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Proteínas Proto-Oncogênicas c-mdm2 , Proteínas de Ligação a RNA/metabolismo , Proteínas Ribossômicas/metabolismo , Neoplasias Gástricas , Proteína Supressora de Tumor p53/metabolismo , Proteínas Reguladoras de Apoptose/genética , Carcinogênese/genética , Linhagem Celular Tumoral , Ensaios de Migração Celular/métodos , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Humanos , Imidazóis/farmacologia , Piperazinas/farmacologia , Proteínas Proto-Oncogênicas c-mdm2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , RNA Interferente Pequeno/análise , Transdução de Sinais/efeitos dos fármacos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Células Tumorais CultivadasRESUMO
A tapered fiber decorated by Ag nanoparticles is prepared as a surface-enhanced Raman scattering (SERS) substrate. There are two key parameters during the preparation process, the fiber cone angle and the density of decorated AgNPs on the fiber tip surface. Their theoretical analysis on the forming mechanism and the optimization process is studied in detail. The tapered fibers with angles from 0.5 to 30° are successfully prepared, with a chemical method in a small tube using a bending interface. AgNPs with different densities are decorated on the surface of the tapered fibers with an electrostatic adsorption method. The optimized tapered fiber SERS probe with an angle of 12° and AgNPs density of 26.67% provides the detection of Rhodamine 6G (R6G) with 10-10 mol/L.
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Atrial fibrillation (AF) and ischemic heart disease (IHD) represent the two most common clinical cardiac diseases, characterized by angina, arrhythmia, myocardial damage, and cardiac dysfunction, significantly contributing to cardiovascular morbidity and mortality and posing a heavy socio-economic burden on society worldwide. Current treatments of these two diseases are mainly symptomatic and lack efficacy. There is thus an urgent need to develop novel therapies based on the underlying pathophysiological mechanisms. Emerging evidence indicates that oxidative DNA damage might be a major underlying mechanism that promotes a variety of cardiac diseases, including AF and IHD. Antioxidants, nicotinamide adenine dinucleotide (NAD+) boosters, and enzymes involved in oxidative DNA repair processes have been shown to attenuate oxidative damage to DNA, making them potential therapeutic targets for AF and IHD. In this review, we first summarize the main molecular mechanisms responsible for oxidative DNA damage and repair both in nuclei and mitochondria, then describe the effects of oxidative DNA damage on the development of AF and IHD, and finally discuss potential targets for oxidative DNA repair-based therapeutic approaches for these two cardiac diseases.
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Fibrilação Atrial/etiologia , Reparo do DNA , Suscetibilidade a Doenças , Isquemia Miocárdica/etiologia , Estresse Oxidativo , Animais , Fibrilação Atrial/metabolismo , Fibrilação Atrial/terapia , Biomarcadores , Núcleo Celular/genética , Núcleo Celular/metabolismo , Gerenciamento Clínico , Expressão Gênica , Humanos , Modelos Biológicos , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/terapia , Miocárdio/metabolismo , Oxirredução , Espécies Reativas de Oxigênio/metabolismoRESUMO
OBJECTIVES: To investigate adaptive strategies of Tupaia belangeri to environmental factors in different populations, 12 locations were selected, including higher and lower altitude areas. RESULTS: Total of 96 and 90 metabolites were annotated in serum and liver respectively, which were mainly concentrated in primary metabolites. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that the locations were divided into two groups in serum metabolites, but each group had a few samples overlap. The samples of each group overlap to some degree in the liver metabolites. The tricarboxylic acid (TCA) cycle occupies a central position in metabolism. The concentrations of TCA intermediates, lipid metabolites and amino acid metabolites were higher in higher altitude areas, and the concentrations of carbohydrate and glycolysis intermediates were higher in lower altitude areas. CONCLUSIONS: Different areas adapted to the changes of environmental and altitude by regulating the concentration of metabolites in serum and liver, and revealed the adaptive mechanism of T. belangeri in different living environments.
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Biomarcadores/sangue , Metaboloma/genética , Metabolômica , Tupaia/sangue , Aminoácidos/sangue , Aminoácidos/genética , Animais , China , Cromatografia Gasosa-Espectrometria de Massas , Lipídeos/sangue , Fígado/metabolismo , Tupaia/genéticaRESUMO
Simple D-type plastic optical fiber (POF) probes (i.e., sensor, reference, and photochemical probes) were created to accurately monitor the progression and phenol degradation of a Chlorella vulgaris biofilm. The sensor and reference probes were used to monitor the biofilm growth (thickness). The sensor probe, which consisted of a D-shaped POF and Canada balsam doped with GeO2 (CBG) coating, was developed to monitor the biofilm growth and change in the liquid-phase composition and its concentration inside the biofilm. The reference probe, which comprised a D-shaped POF, CBG coating, and glass fiber membrane (to separate the liquids from Chlorella vulgaris), was used to measure the response to changes in the liquid phase. A model was developed to demonstrate the accurate measurement of the biofilm thickness. The photochemical POF probe was coupled with a high-permselectivity phenol polymer membrane to monitor the phenol concentration and analyze the degradation time of 50 mg/L phenol with microalgal biofilms. A fixed relationship was obtained between the biofilm sensor output information and biofilm thickness for a biofilm thickness range of 0-290 µm with a periodic supply of 50 mg/L phenol solution. The highest phenol degradation rate occurred at a biofilm thickness of 191-222 µm. The proposed system can be used to investigate microalgal biomass and can provide a promising avenue for research on renewable resources and pollutant degradation.
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Biofilmes/crescimento & desenvolvimento , Tecnologia de Fibra Óptica , Microalgas/metabolismo , Fenol/metabolismo , Tecnologia de Fibra Óptica/instrumentação , Fenol/químicaRESUMO
A photochemical fiber-optic sensor was developed by integrating a plastic optical fiber (POF), polymer membrane, gold mirror, and TiO2-based composite, and was shown to sensitively and selectively detect phenol in aqueous solution. The sensing element consisted of a thinned POF and visible-light-driven SiO2/N-doped TiO2 coating. The gold mirror was used to develop a reflective POF probe. The polymer membrane with high phenol permselectivity was employed to form a micro-channel between the membrane and probe. Our findings highlight the sensor's capability of phenol detection in aqueous solutions with high sensitivity of 0.294×10-3 (mg·L-1)-1, pH immunity ranging from 2.0 to 14.0, and high selectivity with a limit of detection of 30 µg·L-1.
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In this work, we present a luminous-exothermic hollow optical element (LEHOE) that performs spectral beam splitting in the visible spectral range for the enhancement of biofilm growth and activity. The LEHOE is composed of a four-layer structure with a fiber core (air), cladding (SiO2), coating I (LaB6 film), and coating II (SiO2-Agarose-Medium film). To clarify the physical, optical and photothermal conversion properties of the LEHOE, we determined the surface morphology and composition of the coating materials, and examined the luminous intensity and heating rate at the LEHOE surface. The biofilm activity on the biocompatible LEHOE is far greater than that of commercial fibers, and the biofilm weight on the LEHOE is 4.5 × that of the uncoated hollow optical element.
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In this study, a simple wet etching technique is developed by employing aqueous solutions of acetic acid and ultrasonic irradiation for the fabrication of a high-quality plastic optical fiber (POF) sensor. The effects of acetic acid concentration and temperature and ultrasonic power on the etching rate and surface morphology of the etched POFs are investigated. The transmission spectrum and sensitivity of the etched POF sensors are evaluated using glucose solutions. We discovered that the POF sensors, which are fabricated using an aqueous solution of acetic acid with a concentration of 80 vol. % under an ultrasonic power of 130 W and temperature of 25°C, exhibit good light transmission and a high sensitivity of 9.10 [(RIU)(g/L)]-1 in the glucose solutions.
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In this study, we present a novel surgical method that utilizes the ultrasonic bone scalpel (UBS) for the removal of large retrovertebral osteophytes in anterior cervical discectomy and fusion (ACDF) and evaluate its safety and efficacy in comparison to the traditional approach of using high-speed drill (HSD). A total of 56 patients who underwent ACDF for retrovertebral osteophytes were selected. We recorded patients' baseline information, operation time, intraoperative blood loss, complications, JOA and VAS scores, and other relevant data. The mean operation time and the mean intraoperative blood loss in the UBS group were less than those in the HSD group (P < 0.05). Although both groups exhibited considerable improvements in JOA and VAS scores following surgery, there was no statistically significant difference between the two groups (P > 0.05). Additionally, no significant disparities were found in bone graft fusion between the two groups at 6- and 12-months postsurgery. Notably, neither group exhibited complications such as dura tear or spinal cord injury. Our study found that the use of UBS reduced operative time, minimized surgical bleeding, and led to clinical outcomes comparable to HSD in ACDF. This technique offers an effective and safe method of removing large retrovertebral osteophytes.
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Osteófito , Fusão Vertebral , Humanos , Estudos Retrospectivos , Osteófito/cirurgia , Perda Sanguínea Cirúrgica , Ultrassom , Fusão Vertebral/métodos , Resultado do Tratamento , Discotomia/efeitos adversos , Discotomia/métodos , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgiaRESUMO
Melatonin (MEL) is an indole hormone synthesized and secreted by the pineal gland at night, which is involved in the regulation of body mass and thermogenesis in small mammals. To test the effects of exogenous MEL on body mass and thermogenic ability in two different red-backed vole (Eothenomys miletus) populations from two different regions (Kunming [KM] and Dali [DL]) with different annual variation in climatic variables, such as temperature, sunshine and rainfall. we traced the changes of energy balance in E. miletus from KM and DL, which were placed at 25 ± 1°C with photoperiod of 12 L:12 D, intraperitoneal injection of MEL was performed daily for 28 days. The results showed that body mass and food intake were significantly decreased, while resting metabolic rate (RMR) and nonshivering thermogenesis (NST) were significantly increased after MEL injection; Contents of total protein, mitochondrial protein, the activities of cytochrome C oxidase (COX) and α-glycerophosphate oxidase (α-PGO) in liver and brown adipose tissue (BAT) were enhanced; the activity of thyroxin 5'-deiodinase (T4 5'-DII) and uncoupling protein 1 (UCP1) in BAT were also increased. Serum leptin, triiodothyronine (T3 ) levels and T3 /T4 ratio were significantly increased, thyroxine (T4 ) levels was significantly decreased. Moreover, body mass and food intake in E. miletus from KM were higher than those from DL, but RMR and NST were lower than those from DL. Changes of body mass, food intake and thermogenic activity of KM were higher than those of DL when exposed to injection of MEL, indicating that E. miletus in KM were more sensitive to MEL. Furthermore, MEL was involved in the regulation of body mass and thermogenesis in E. miletus between KM and DL.
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Melatonina , Animais , Melatonina/farmacologia , Arvicolinae/fisiologia , Termogênese/fisiologia , Tri-Iodotironina/farmacologia , MamíferosRESUMO
Physiological adaptation of tree shrews (Tupaia belangeri) to changing environmental temperature has been reported in detail. However, the T. belangeri origin (mainland or island), population history, and adaptation to historical climate change remain largely unknown or controversial. Here, for the first time, we sequenced the simplified genome of 134 T. belangeri individuals from 12 populations in China and further resequenced one individual from each population. Using population genomic approaches, we first observed considerable genetic variation in T. belangeri. Moreover, T. belangeri populations formed obvious genetic structure and reflected different demographic histories; they generally exhibited high genetic diversity, although the isolated populations had relatively low genetic diversity. The results presented in this study indicate that T. b. modesta and T. b. tonquinia were separated recently and with a similar population dynamics. Second, physical barriers rather than distance were the driving factors of divergence, and environmental heterogeneity may play an important role in genetic differentiation in T. belangeri. Moreover, our analyses highlight the role of historical global climates in the T. belangeri population dynamics and indicate that the decrease of the T. belangeri population size may be due to the low temperature. Finally, we identified the olfaction-associated adaptive genes between different altitude populations and found that olfactory-related genes of high-altitude populations were selectively eliminated. Our study provides demographic history knowledge of T. belangeri; their adaption history offers new insights into their evolution and adaptation, and provides valuable baseline information for conservation measures.
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Tupaia , Tupaiidae , Animais , Tupaia/genética , Metagenômica , Aclimatação , ChinaRESUMO
An important objective of evolutionary biology has always been to grasp the evolutionary and genetic processes that contribute to speciation. The present work provides the first detailed account of the genetic and physiological adaptation to changing environmental temperatures as well as the reasons causing intraspecific divergence in the Eothenomys miletus from the Hengduan Mountain (HM) region, one of the biodiversity hotspots. One hundred sixty-one E. miletus individuals from five populations in the HM region had their reduced-representation genome sequenced, and one additional individual from each community had their genomes resequenced. We then characterized the genetic diversity and population structure of each population and compared the phenotypic divergence in traits using neutral molecular markers. We detected significant phenotypic and genetic alterations in E. miletus from the HM region that were related to naturally occurring diverse habitats by combining morphometrics and genomic techniques. There was asymmetric gene flow among the E. miletus populations, indicating that five E. miletus populations exhibit an isolation-by-island model, and this was supported by the correlation between F ST and geographic distance. Finally, P ST estimated by phenotypic measures of most wild traits were higher than differentiation at neutral molecular markers, indicating directional natural selection favoring different phenotypes in different populations must have been involved to achieve this much differentiation. Our findings give information on the demographic history of E. miletus, new insights into their evolution and adaptability, and literature for studies of a similar nature on other wild small mammals from the HM region.
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BACKGROUND: Circulating tumor DNA (ctDNA)-based minimal residual disease (MRD) detection, which can identify disease relapse ahead of radiological imaging, has shown promising performance. The objective of this study was to develop and validate OriMIRACLE S (Minimal Residual Circulating Nucleic Acid Longitudinal Detection in Solid Tumor), a highly sensitive and specific tumor-informed assay for MRD detection. METHODS: Tumor-specific somatic single nucleotide variants (SNVs) were identified via whole exome sequencing of tumor tissue and matched germline DNA. Clonal SNVs were selected using the OriSelector algorithm for patient-specific, multiplex PCR-based NGS assays in MRD detection. Plasma-free DNA from patients with gastrointestinal tumors prior to and following an operation, and during monitoring, were ultradeep sequenced. RESULTS: The detection of three positive sites was sufficient to achieve nearly 100% overall sample level sensitivity and specificity and was determined by calculating binomial probability based on customized panels containing 21 to 30 variants. A total of 127 patients with gastrointestinal tumors were enrolled in our study. Preoperatively, MRD was positive in 18 of 26 patients (69.23%). Following surgery, MRD was positive in 24 of 82 patients (29.27%). The positivity rate for MRD was 33.33% (n = 18) for gastric adenocarcinoma and 32.26% (n = 62) for colorectal cancer. Twenty (20) of 59 patients (34.48%) experienced a change in MRD status over the monitoring period. Patients 8 and 31 responded to 3 cycles of systemic therapy, after which levels for all ctDNA dropped below the detection limit. Patient 53 was an example of using MRD to predict tumor metastasis. Patient 55 showed a weak response to treatments first and respond to new systemic therapy after tumor progression. CONCLUSION: Our study identified a sensitive and specific clinical detection method for low frequency ctDNA, and explored the detection performance of this technology in gastrointestinal tumors.
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Carcinoma , DNA Tumoral Circulante , Neoplasias Gastrointestinais , Humanos , DNA Tumoral Circulante/genética , Neoplasia Residual/genética , Recidiva Local de Neoplasia , Neoplasias Gastrointestinais/diagnóstico , Neoplasias Gastrointestinais/genéticaRESUMO
Environmental factors, such as photoperiod and temperature, play an important role in the regulation of an animal's physiology and behavior. In the present study, we examined the effects of short photoperiod (SD, 8L:16D) on body mass as well as on several physiological, hormonal, and biochemical measures indicative of thermogenic capacity, to test our hypothesis that short photoperiod stimulates increases thermogenic capacity and energy intake in tree shrews. At the end, these tree shrews (SD) had a significant higher body mass, energy intake, cytochrome C oxidase (COX) activity and uncoupling protein-1 (UCP1) content, serum tri-iodothyronine (T(3)) and thyroxine (T(4)) compared to LD (16L:8D) tree shrews. However, there were no significant differences in serum leptin and melatonin between the two groups. Together, these data suggest tree shrews employ a strategy of maximizing body growth and increasing energy intake in response to cues associated with short photoperiod.
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Adaptação Fisiológica , Leptina/sangue , Melatonina/sangue , Fotoperíodo , Tecido Adiposo Marrom/enzimologia , Tecido Adiposo Marrom/fisiologia , Animais , Metabolismo Basal , Composição Corporal , Respiração Celular , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Ingestão de Energia , Canais Iônicos/metabolismo , Masculino , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Tamanho do Órgão , Estações do Ano , Termogênese , Tupaiidae , Proteína Desacopladora 1 , Aumento de PesoRESUMO
Environmental cues play important roles in the regulation of an animal's physiology and behavior. The purpose of the present study was to test the hypothesis that ambient temperature is a cue to induce adjustments in body mass, energy intake and thermogenic capacity, associated with changes in serum leptin levels in tree shrews (Tupaia belangeri). We found that tree shrews increased basal metabolic rate (BMR), energy intake and subsequently showed a significant decrease in body mass after being returned to warm ambient temperature. Uncoupling protein 1 (UCP1) content in brown adipose tissue (BAT) increased during cold acclimation and reversed after rewarming. The trend of energy intake increased during cold acclimation and decreased after rewarming; the trend of energy intake during cold acclimation was contrary to the trend of energy intake during rewarming. Further, serum leptin levels were negatively correlated with body mass. Together, these data supported our hypothesis that ambient temperature was a cue to induce changes in body mass and metabolic capacity. Serum leptin, as a starvation signal in the cold and satiety signal in rewarming, was involved in the processes of thermogenesis and body mass regulation in tree shrews.
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Aclimatação , Peso Corporal/fisiologia , Metabolismo Energético , Termogênese , Tupaiidae/metabolismo , Tecido Adiposo Marrom/anatomia & histologia , Animais , Metabolismo Basal , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Ingestão de Energia , Canais Iônicos/metabolismo , Leptina/sangue , Fígado/anatomia & histologia , Mitocôndrias Hepáticas/enzimologia , Mitocôndrias Hepáticas/metabolismo , Proteínas Mitocondriais/metabolismo , Tupaiidae/fisiologia , Proteína Desacopladora 1RESUMO
Global warming is becoming the future climate trend and will have a significant impact on small mammals, and they will also adapt at the physiological levels in response to climate change, among which the adaptation of energetics is the key to their survival. In order to investigate the physiological adaptation strategies in Tupaia belangeri affected by the climate change and to predict their possible fate under future global warming, we designed a metabonomic study in T. belangeri between two different places, including Pianma (PM, annual average temperature 15.01°C) and Tengchong (TC, annual average temperature 20.32°C), to analyze the differences of liver metabolite. Moreover, the changes of resting metabolic rate, body temperature, uncoupling protein 1content (UCP1) and other energy indicators in T. belangeri between the two places were also measured. The results showed that T. belangeri in warm areas (TC) reduced the concentrations of energy metabolites in the liver, such as pyruvic acid, fructose 6-phosphate, citric acid, malic acid, fumaric acid etc., so their energy metabolism intensity was also reduced, indicating that important energy metabolism pathway of glycolysis and tricarboxylic acid cycle (TCA) pathway reduced in T. belangeri from warmer habitat. Furthermore, brown adipose tissue (BAT) mass, UCP1 content and RMR in TC also decreased significantly, but their body temperature increased. All of the results suggested that T. belangeri adapt to the impact of warm temperature by reducing energy expenditure and increasing body temperature. In conclusion, our research had broadened our understanding of the physiological adaptation strategies to cope with climate change, and also provided a preliminary insight into the fate of T. belangeri for the future global warming climate.
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In order to investigate the effects of different areas on intestinal bacterial diversity and body mass regulation in Eothenomys miletus from Hengduan mountain regions, and to explore the community structure and diversity of intestinal microflora and their role in body mass regulation. E. miletus was collected from five areas including Deqin (DQ), Xianggelila (XGLL), Lijiang (LJ), Jianchuan (JC), and Dali (DL), we used 16S rRNA sequencing technology combined with physiological and morphological methods to study the intestinal microbiota diversity, abundance and community structure of the intestinal bacteria in winter, and to explore the influence of geographical factors, physiological indicators including food intake, resting metabolic rate (RMR), non-shivering thermogenesis (NST), neuropeptide Y (NPY), Agouti-Related Protein (AgRP), proopiomelanocortin (POMC), cocaine and amphetamine regulated transcription peptide (CART), and morphological indicators including body mass, body length and other nine indicators on the intestinal microflora diversity in E. miletus. The results showed that there were significant differences in metabolic indexes such as RMR, NST, NPY, AgRP, and morphological indexes such as body length, tail length and ear length among the five regions. Bacterial community in intestinal tract of E. miletus mainly includes three phyla, of which Firmicutes is the dominant phyla, followed by Bacteroidetes and Tenericutes. At the genus level, the dominant bacterial genera were S24-7(UG), Clostridiales (UG), and Lachnospiraceae (UG), etc. α diversity of intestinal microorganisms in DL and JC were significantly different from that in the other three regions. Genera of intestinal microorganisms in DL and JC were also the most. Moreover, Bacteroides, Ruminococcus, and Treponema could affect energy metabolism in E. miletus, which were closely related to the environment in which they lived. All of these results indicated that different areas in Hengduan Mountain had certain effects on the structure of intestinal microbial community in E. miletus, which were responded positively to changes in food abundance and other environmental factors. Furthermore, Firmicutes and Bacteroidetes play an important role in the body mass regulation in E. miletus.
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Leptin is a hormone mainly synthesized and secreted by white adipose tissue (WAT), which regulates various physiological processes. To investigate the role of leptin in energy balance and thermoregulation in Eothenomys miletus, voles were randomly divided into leptin-injected and PBS-injected groups and placed at 25°C ± 1°C with a photoperiod of 12 L:12 D. They were housed under laboratory conditions for 28 days and compared in terms of body mass, food intake, water intake, core body temperature, interscapular skin temperature, resting metabolic rate (RMR), nonshivering thermogenesis (NST), liver and brown adipose tissue (BAT) thermogenic activity, and serum hormone levels. The results showed that leptin injection decreased body mass, body fat, food intake, and water intake. But it had no significant effect on carcass protein. Leptin injection increased core body temperature, interscapular skin temperature, resting metabolic rate, non-shivering thermogenesis, mitochondrial protein content and cytochrome C oxidase (COX) activity in liver and brown adipose tissue, uncoupling protein 1 (UCP1) content and thyroxin 5'-deiodinase (T45'-DII) activity in brown adipose tissue significantly. Serum leptin, triiodothyronine (T3), thyrotropin-releasing hormone (TRH) and corticotropin-releasing hormone (CRH) concentrations were also increased significantly. Correlation analysis showed that serum leptin levels were positively correlated with core body temperature, body mass loss, uncoupling protein 1 content, thyroxin 5'-deiodinase activity, nonshivering thermogenesis, and negatively correlated with food intake; thyroxin 5'-deiodinase and triiodothyronine levels were positively correlated, suggesting that thyroxin 5'-deiodinase may play an important role in leptin-induced thermogenesis in brown adipose tissue. In conclusion, our study shows that exogenous leptin is involved in the regulation of energy metabolism and thermoregulation in E. miletus, and thyroid hormone may play an important role in the process of leptin regulating energy balance in E. miletus.