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Background It is widely accepted that high mobility group box 1 (HMGB1) is a key inflammatory mediator in the pathogenesis of sepsis. HMGB1 promotes inflammation during sterile and infectious damage and plays a crucial role in disease development. Mobilization from the nucleus to the cytoplasm is the first important step in the release of HMGB1 from activated immune cells. Sirtuin 2 (SIRT2) is a member of the sirtuin family, nicotinamide adenine dinucleotide+-dependent deacylases, which participates in modulation of cell cycle control and neurodegeneration Methods A sepsis mouse modle was established using C57BL/6 and SIRT2-/- mice by LPS induction. Assessment of microglia polarization and pro-inflammatory markers, such as tumor necrosis factor IL-1β, IL-6, and HMGB1. Immunofluorescent staining and western blot analysis were used to detect HMGB1 nuclear translocation. The impact of SIRT2 deacetylase activity on HMGB1 acetylation and the interaction between HMGB1 and SIRT2. Results The results of our study showed that HMGB1 was reduced after SIRT2 knockdown. SIRT2 overexpression increased HMGB1. SIRT2 affected the translocation of HMGB1 from the nucleus to the cytoplasm. In addition, SIRT2 interacts with HMGB1 and deacetylates the K43 lysine sites. In vivo, a decrease in HMGB1 due to the absence of SIRT2. SIRT2 depletion increased HMGB1 ubiquitination and had an impact on the nucleoplasmic shuttle. Conclusion These discoveries are the first to shed light on the SIRT2 nucleoplasmic shuttle, which influences HMGB1 and its degradation, hence revealing novel therapeutic targets and avenues for sepsis treatment.

Objective: O-GlcNAcylation, a nutrient sensing pathway involved in a myriad of cellular processes, it plays a key role in metabolism homeostasis. Cold stress disturbs cellular metabolic and energy homeostasis and is one of the causes of stress-induced illnesses. It is reported that O-GlcNAcylation has been shown to be upregulated during cold stress. Skeletal muscle and its residential mitochondria play an important role in maintaining the metabolic homeostasis of the whole body. Nevertheless, the mechanism of which O-GlcNAcylation adapts to cold stress in skeletal muscle remains unknown.Methods: To characterize the effect of O-GlcNAcylation on mouse skeletal muscle during cold exposure, this research investigated the effects of skeletal muscle structure, function, mitochondrial homeostasis, the deacetylation activity of SIRT1, acetylation expression levels and oxidative stress levels in Ogt mKO mice by cold exposure (4 ℃, 3 h a day, for 1 week). In order to understand the mechanism of O-GlcNAcylation on skeletal muscle homeostasis under cold stress, C2C12 cells was used as a model in vitro. C2C12 cells were treated with OGT inhibitor (Alloxan) and OGA inhibitor (Thiamet G) to decrease and enhance O-GlcNAcylation signals in mild hypothermia, respectively, and then recapitulated in vivo phenotype. Finally, the interaction between OGT and SIRT1 was demonstrated, and the O-GlcNAcylation of SIRT1 played an important role in the imbalance of skeletal muscle homeostasis induced by cold exposure.Results: Results showed that in addition to enhance cold stress-induced mitochondrial abnormalities, cell senescence, and collagen accumulation, Ogt deficiency was found to induce autophagy, mitophagy and oxidative stress level in skeletal muscle during cold exposure. And the results showed that the expression level of SIRT1 was reduced, concomitant with increased expression and acetylation of FoxO1 in wild type mice challenged with cold stress. Because the depletion of Ogt further reduced SIRT1, and increased FoxO1 in muscle challenged with cold stress. Furthermore, this study showed that OGT had a physiological interaction with SIRT1. Thr160 and Ser161 sites of SIRT1 could be O-GlcNAcylation, and overexpression of SIRT1 saved mitochondrial defects in C2C12 cells caused by cold exposure.Conclusion: Cold stress can cause skeletal muscle damage, and O-GlcNAcylation of SIRT1 is an important protection mechanisms of skeletal muscle’s adaptation to cold stress.

The spiny frog Quasipaa robertingeri is endemic to a narrow region of southwest China and its taxonomic validation is still controversial. Based on COI gene sequences of 110 individuals from seven ...

A new species of the genus Tylototriton sensu lato from Tongzi County, Guizhou Province, China was described. Molecular phylogenetic analyses based on mitochondrial 16S and ND2 gene sequences indicated the new species as the most closely related species of T. dabienicus in Henan. The new species could be identified from its congeners by a combination of the following morphological characters: (1) body size medium (TOL 120.5–135.1 mm and SVL 61.1–65.9 mm in males, and TOL 123.5–127.6 mm and SVL 66.7–69.2 mm in females); (2) gular fold present; (3) the tail length shorter than the snout-vent length; (4) the distal ends and ventral surfaces of digits, peripheral area of cloaca, and the lower margin of tail orange; (5) the distal tips of the limbs greatly overlapping when the fore and hind limbs being pressed along the trunk; (6) fingertips reaching to the level beyond the snout when the forelimbs being stretched forward; (7) nodule-like warts on body sides continuous and no obvious. The new species is known only from the montane forests of Huanglian Nature Reserve, Tongzi County, Guizhou Province, China. We recommend the new species to be listed as Critically Endangered.

A new species of the genus Megophrys is described from Guizhou Province, China. Molecular phylogenetic analyses based on mitochondrial DNA indicated the new species as a clade clustered into the Megophrys clade. The new species can be distinguished from its congeners by a combination of the following characters: body size moderate (SVL 40.0–45.5 mm in males and 48.9–51.2 mm in females); vomerine teeth absent; tongue not notched behind; tympanum distinctly visible, oval; a small horn-like tubercle at the edge of each upper eyelid; two metacarpal tubercles in hand; toes with rudimentary webbing; heels overlapping when thighs are positioned at right angles to the body; tibiotarsal articulation reaching the level of mid-eye when leg stretched forward; in breeding males, an internal single subgular vocal sac present and brownish nuptial pads, made up of black nuptial spines, present on the dorsal base of the first two fingers.

A new species of the Asian leaf litter toad genusLeptobrachellafrom Guizhou Province, China is described based on molecular phylogenetic analyses, morphological comparisons, and bioacoustics data. Phylogenetic analyses based on the mitochondrial 16S rRNA gene sequences supported the new species as an independent clade nested into theLeptobrachellaclade and sister toL. bijie. The new species could be distinguished from its congeners by a combination of the following characters: small body size (SVL 30.8–33.4 mm in seven adult males, and 34.2 mm in one adult female); dorsal skin shagreened, some of the granules forming longitudinal short skin ridges; tympanum distinctly discernible, slightly concave; internasal distance longer than interorbital distance; supra-axillary, femoral, pectoral and ventrolateral glands distinctly visible; absence of webbing and lateral fringes on fingers; toes with rudimentary webbing and shallow lateral fringes, relative finger lengths II < IV < I < III; heels overlapped when thighs are positioned at right angles to the body; and tibia-tarsal articulation reaches the tympanum.

Cold stimulation reduces the quality of animal products and increases animal mortality, causing huge losses to the livestock industry in cold regions. Long non-coding RNAs (lncRNAs) take part in many biological processes through transcriptional regulation, intracellular material transport, and chromosome remodeling. Although cold stress-related lncRNAs have been reported in plants, no research is available on the characteristic and functional analysis of lncRNAs after cold stress in rats. Here, we built a cold stress animal model firstly. Six SPF male Wistar rats were randomly divided to the acute cold stress group (4 °C, 12 h) and the normal group (24 °C, 12 h). lncRNA libraries were constructed by high-throughput sequencing (HTS) using rat livers. 2,120 new lncRNAs and 273 differentially expressed (DE) lncRNAs were identified in low temperature environments. The target genes of DElncRNA were predicted by cis and trans, and then functional and pathway analysis were performed to them. GO and KEGG analysis revealed that lncRNA targets were mainly participated in the regulation of nucleic acid binding, cold stimulation reaction, metabolic process, immune system processes, PI3K-Akt signaling pathway and pathways in cancer. Next, a interaction network between lncRNA and its targets was constructed. To further reveal the mechanism of cold stress, DElncRNA and DEmRNA were extracted to reconstruct a co-expression sub-network. We found the key lncRNA MSTRG.80946.2 in sub-network. Functional analysis of key lncRNA targets showed that targets were significantly enriched in fatty acid metabolism, the PI3K-Akt signaling pathway and pathways in cancer under cold stress. qRT-PCR confirmed the sequencing results. Finally, hub lncRNA MSTRG.80946.2 was characterized, and verified its relationship with related mRNAs by antisense oligonucleotide (ASO) interference and qRT-PCR. Results confirmed the accuracy of our analysis. To sum up, our work was the first to perform detailed characterization and functional analysis of cold stress-related lncRNAs in rats liver. lncRNAs played crucial roles in energy metabolism, growth and development, immunity and reproductive performance in cold stressed rats. The MSTRG.80946.2 was verified by network and experiments to be a key functional lncRNA under cold stress, regulating ACP1, TSPY1 and Tsn.

The Music frog genus Nidirana is widely distributed in East and South Asia. Here, a new species of the genus is described from southwestern China. Phylogenetic analyses based on the mitochondrial 16S rRNA and COI gene sequences supported the new species as a clade closely related to N. leishanensis, N. hainanensis, N. chapaensis, N. daunchina, and N. yaoica. The new species could be distinguished from its congeners by a combination of the following characters: body of medium size (SVL 41.2–43.5 mm in males and 44.7 mm in female); lateroventral groove only present on toes; relative finger lengths: II < IV < I < III; three metatarsal tubercles on palm; heels overlapping when hindlimbs flexed at right angles to axis of body; tibiotarsal articulation reaching the level of eye when leg stretched forward; a pair of subgular internal vocal sacs at corners of throat in male; nuptial pad present on the inner side of base of fingers I in breeding male; tadpole labial tooth row formula with 1:1+1/1+1:2; in males, the advertisement call contains two kinds of notes and one call contains 2–6 repeated regular notes.

Stress induces many different sex-specific physiological and psychological responses during adolescence. Although the impact of certain brain stressors has been reported in the literature, the influence of cold stress on the mechanisms underlying hippocampal neurotransmitter disorder and neuroinflammation remain unstudied. Adolescent male and female C57BL/6 mice were exposed to 4 °C temperatures, 3 h per day for 1 week. Serum CORT and blood gas analysis was then used to assess body status. Using western blotting, immunofluorescence and immunohistochemistry we also assessed glial cell number and microglial activation, as well as inflammatory cytokine levels and related protein expression levels. The phenomena of excessive CORT, microglial activation, increased acetylate-HMGB1 levels, NF-κB signaling pathway activation, pro-inflammatory cytokine release, neuronal apoptosis and neurotransmitter disorder were demonstrated in mouse hippocampal tissue following cold exposure. We believe that these phenomena are mediated by the HMGB1/TLR4/NFκB pathway. Finally, the male inflammatory response in hippocampal tissue was more severe and the influence of cold exposure on neurotransmitter was greater in females.

Stress induces many non-specific inflammatory responses in the mouse brain, especially during adolescence. Although the impact of stress on the brain has long been reported, the effects of cold stress on hippocampal neuroinflammation in adolescent mice are not well understood; furthermore, whether these effects are gender specific are also not well established. Adolescent male and female C57BL/6 mice were exposed to 4 °C temperatures for 12 h, after which behavior was assessed using the open field test. Using western blotting and immunohistochemistry we also assessed glial cell numbers and microglial activation, as well as inflammatory cytokine levels and related protein expression levels. We found that in mice subjected to cold stress: 1) There were significant behavioral changes; 2) neuronal nuclei densities were smaller and total cell numbers were significantly decreased; 3) nuclear factor (NF)-κB and phosphorylated AKT were upregulated; 4) pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-α were also upregulated; and 5) microglia were activated, while glial fibrillary acid protein and ionized calcium-binding adapter molecule 1 protein expression increased. Taken together, these results indicate that cold stress induces pro-inflammatory cytokine upregulation that leads to neuroinflammation and neuronal apoptosis in the hippocampi of adolescent mice. We believe that these effects are influenced by a GABAB/Rap1B/AKT/NF-κB pathway. Finally, male mice were more sensitive to the effects of cold stress than were female mice.

A new species of the genus Megophrys is described from Guizhou Province, China. Molecular phylogenetic analyses based on mitochondrial DNA and nuclear DNA sequences all strongly supported the new species as an independent clade sister to M. minor and M. jiangi. The new species could be distinguished from its congeners by a combination of the following characters: body size moderate (SVL 43.4–44.1 mm in males, and 44.8–49.8 mm in females; vomerine teeth absent; tongue not notched behind; a small horn-like tubercle at the edge of each upper eyelid; tympanum distinctly visible, rounded; two metacarpal tubercles on palm; relative finger lengths II < I < V < III; toes without webbing; heels overlapping when thighs are positioned at right angles to the body; tibiotarsal articulation reaching the level between tympanum and eye when leg stretched forward; in breeding males, an internal single subgular vocal sac in male, and the nuptial pads with black spines on dorsal surface of bases of the first two fingers.

Chronic stress can damage homeostasis and induce various primary diseases. Although chronic cold stress is becoming an increasing problem for people who must work or live in extreme environments, risk-induced diseases in the central nervous system remain unstudied. Male C57BL/6 mice were exposed to an environment of 4 &deg, C, 3 h per day for 1, 2, and 3 weeks and homeostasis in the hippocampus and neuronal apoptosis were evaluated by Western blotting, immunohistochemistry, TdT-mediated dUTP Nick-End Labeling (TUNEL) staining, and immunofluorescence. The phenomena of oxidation stress, MAPK signaling pathway activation, anti-oxidation protein release, neuronal apoptosis increases, and neuronal proliferation inhibition were demonstrated in the CA1 and CA3 regions of mouse hippocampal tissues following cold exposure. We speculated that these phenomena were mediated by the MAPK pathway and were closely linked with oxidative stress in the hippocampus. This study provides novel concepts regarding neurodegenerative diseases, suggesting that chronic cold stress may be a critical factor to induce neurodegenerative diseases.

The Music frog genus Nidirana is widely distributed in East and South Asia. Here, a new species of the genus is described from southwestern China. Phylogenetic analyses based on the mitochondrial 16S rRNA and COI gene sequences supported the new species as a clade closely related to N. leishanensis, N. hainanensis, N. chapaensis, N. daunchina, and N. yaoica. The new species could be distinguished from its congeners by a combination of the following characters: body of medium size (SVL 41.2–43.5 mm in males and 44.7 mm in female); lateroventral groove only present on toes; relative finger lengths: II < IV < I < III; three metatarsal tubercles on palm; heels overlapping when hindlimbs flexed at right angles to axis of body; tibiotarsal articulation reaching the level of eye when leg stretched forward; a pair of subgular internal vocal sacs at corners of throat in male; nuptial pad present on the inner side of base of fingers I in breeding male; tadpole labial tooth row formula with 1:1+1/1+1:2; in males, the advertisement call contains two kinds of notes and one call contains 2–6 repeated regular notes.

Cold stress can induce neuroinflammation in the hippocampal dentate gyrus (DG), but the mechanism underlying neuronal apoptosis induced by cold stress is not well-understood. To address this issue, male and female C57BL/6 mice were exposed to a temperature of 4 &deg, C for 3 h per day for 1 week, and glial cell activation, neuronal apoptosis, and neuroinflammation were evaluated by western blotting, immunofluorescence, terminal deoxynucleotidyl transferase 2&rsquo, deoxyuridine 5&rsquo, triphosphate (dUTP) nick end labeling, Nissl staining, and immunohistochemistry. Additionally, BV2 cells were treated with different concentrations of cortisol (CORT) for 3 h to mimic stress and molecular changes were assessed by western blotting, immunofluorescence, and co-immunoprecipitation. We found that excess CORT activated glial cells and increased neuroinflammation in the DG of mice exposed to cold temperatures, which was associated with increased acetylation and nuclear factor-&kappa, B signaling. These effects were mediated by the acetylation of lysine 9 of histone 3 and lysine 310 of p65, which resulted in increased mitogen-activated protein kinase phosphorylation, nuclear translocation of p65, microglia activation, and acetylation of high-mobility group box 1. Neuroinflammation was more severe in male compared to female mice. These findings provide new insight into the mechanisms of the cold stress response, which can inform the development of new strategies to combat the effects of hypothermia.

Prenatal cold stress is one of the earliest factors affecting mammalian health, and is associated with neonatal growth retardation and immune dysfunction, thus increasing disease susceptibility. The mechanisms underlying these observations remain unclear; hence, the objective of this study was to elucidate placental responses to cold stress. 60 maternal rats were randomly allocated to either stressed (n = 30) or non-stressed (control, n = 30) treatment conditions and 30 pubs (n=15) were used for the pups analysis. We found that maternal exposure to cold stress resulted in decreased body temperature, increased food intake without body weight gain, and a high level of plasma corticosterone (CORT) between gestational day (GD) 14 and GD21. In addition, gestation cold stress induced the placental expression of heat shock protein 70 (HSP70), IκBα, glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2), interferon (IFN) regulatory factor 3 (IRF3), Caspase-3 proteins and altered the ratio of B-cell lymphoma-extra large (Bcl-xL) to Bcl-associated x (Bax) proteins on gestational GD15, GD17, GD19, and GD21, also resulted in the production of interleukin (IL)-1β. Next, gestational cold stress provoked a decrease in plasma GH levels of 21-day-old offspring, and the body weights of offspring were have no differences from postnatal day (PD) 1–21. Taken together, our results indicate that gestational cold stress induces placental apoptosis and the activation of NF-kB via HSP70/TLR4/NF-κB signaling pathways in the placenta, these changes may affect placental function and fetus development.

A new species of the microhylid frog genus Microhyla is described from the Fanjing Mountain of Guizhou Province, China. Phylogenetic analyses based on DNA sequences of the the mitochondrial 12S rRNA, 16S rRNA and COI genes supported the new species as an independent lineage, closely related to M. beilunensis, M. mixtura and M. okinavensis. The uncorrected genetic distance on 16S rRNA gene between the new species and its closest congeners M. beilunensis, M. mixtura and M. okinavensis are 3.5%, 4.6% and 4.6% respectively. The new species is distinguished from its congeners by a combination of the following morphological characters: (1) body of medium size (SVL 19.0–22.7 mm in males and 22.5–23.0 mm in females); (2) disk and dorsal median longitudinal groove on finger tips absent; (3) toe with rudimentary webbing at base; (4) disk with dorsal median longitudinal groove present at toe tips except for the toe I; (5) two metatarsal tubercles on palm; (6) tibiotarsal articulation reaching the level between eye to nostril when leg stretched forward; (7) a distinct V-shaped white stripe on the upper midsection. The new species is known only from the type locality, and thus the finding of it has contributed to the endemic species diversity of China. It is needed to take strategy to protect the species and habitats due to the increased threats of environmental changes.

The Asian music frog genus Nidirana is widely distributed in East and Southeastern Asia. Systematic profiles of the group remain on debate, and cryptic species are expected especially in the species with wide distributional range. Here, we describe a new species of the genus from Southwestern China. Phylogenetic analyses based on mitochondrial DNA and nuclear DNA supported the new species as an independent clade nested into the Nidirana clade and sister to N. hainanensis. Morphologically, the new species could be distinguished from its congeners by a combination of the following characters: a large body size in males (SVL > 49 mm); the presence of lateroventral grooves both on fingers and toes; relative finger lengths: II 1/3 II 2 –22/3 III 31/2 –32/3 IV 32/3 –3V. The findings provided a better knowledge on phylogenetic assignments of the genus Nidirana, and indicated future deeper investigations necessarily for exploring systematic settings of the group.

Maternal stress is a key risk factor in the development of offspring. We previously identified prenatal cold stress-induced anxiety-like behavior reduced in the offspring of rats along with negative feedback regulation from the maternal hippocampus on the hypothalamic-pituitary-adrenal (HPA) axis during prenatal cold stress. However, the precise function of the maternal hypothalamus response to cold stress during late pregnancy in rats has not yet been determined. Therefore, we examined proteins in the hypothalamus that respond to aldosterone, neurodevelopment, inflammation and apoptosis. Our results show that prenatal cold stress induced the expression of mineralocorticoid receptors (MR) and 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2), suggesting prenatal cold stress may promote the elevation of aldosterone levels in the hypothalamus. Remarkably, increased expression of brain derived neurotrophic factor (BDNF) helped to replenish intracellular peptidergic stores and ensure homeostatic balance during prenatal cold stress. Furthermore, prenatal cold stress reduced the expression of c-Fos via STAT3 and ERK1/2 pathways in the hypothalamus. Moreover, prenatal cold stress induced NF-κB phosphorylation at Ser536, then promoted the expression of inducible nitric oxide synthase (iNOS) and induced an apoptosis-related protein response. Together, this study confirms that changes in the maternal hypothalamus during cold stress in late pregnancy are directly reflective of the response of the HPA to cold stress and demonstrates how the hypothalamus coordinates cold stress. We suggest mechanisms which might explain how these states might be linked with an abnormal stress response.

Quinalizarin, a bioactive and highly selective compound, is known to promote apoptosis in colon and lung cancer cells. However, studies evaluating quinalizarin-induced apoptosis in melanoma cells have not been conducted. In the present study, we investigated the underlying mechanisms of antimelanoma activity of quinalizarin in human melanoma A375 cells. The MTT assay and Trypan blue staining were used to evaluate the cell viability. The flow cytometry was used to detect cell cycle, apoptosis and reactive oxygen species (ROS). Western blot was used to detect the expression of cell cycle and apoptosis-related proteins, MAPK, and STAT3. The results revealed a significant dose and time dependent effect of quinalizarin on inhibiting proliferation in three kinds of human melanoma cells, and had no significant toxic effects on normal cells. Moreover, quinalizarin triggered G2/M phase cell arrest by modulating the protein expression levels of CDK 1/2, cyclin A, cyclin B, p21 and p27, and induced apoptosis by down-regulating the antiapoptotic protein Bcl-2 and upregulating the proapoptotic protein BAD, leading to the activation of caspase-3 and PARP in the caspase cascade in A375 cells. Quinalizarin treatment led to apoptosis of A375 cells via activation of MAPK and inhibition of STAT3 signaling pathways. In addition, quinalizarin increased the level of ROS, but ROS scavenger NAC inhibited quinalizarin-induced apoptosis by regulating MAPK and STAT3 signaling pathways. In summary, quinalizarin induces cell cycle arrest and apoptosis via ROS-mediated MAPK and STAT3 signaling pathways in human melanoma A375 cells, and quinalizarin may be used as a novel and effective antimelanoma therapeutic.

Stress induces many non-specific responses in the hippocampus, especially during adolescence. Low environmental temperature is known to induce stress, but its influence on the hippocampus, especially in adolescent mice is not clear. We compared apoptotic-related protein levels and MAPK signaling pathway activation in hippocampal neurons of adolescent mice under low temperature conditions (4 °C for 12 h) with western blotting and immunohistochemistry. Western bolt results demonstrated that the levels of phospho-JNK, phospho-p38, and cleaved-caspase 3 significantly increased, while the ratio of Bcl-XL/Bax decreased, in the cold stress group. The results of immunohistochemistry (IHC) and Nissl staining demonstrated that the protein optical density of caspase 3 increased and Nissl bodies decreased in the cold stress group compared with controls. Thus, we conclude that cold exposure initiates activation of the MAPK signaling pathway and subsequently induces the upregulation of pro-apoptotic proteins in the hippocampi of adolescent mice. Overall our study reveals the relationship between cold stress and apoptosis in adolescent mice.

Stress is a nonspecific response to adverse circumstances and chronic stress can destroy homeostasis, leading to various primary diseases. Although chronic cold stress is becoming increasingly important for individuals living or working in extreme environments, the risk of associated disorders of the central nervous system remains unstudied. Here, male C57BL/6 mice were exposed to a temperature of 4 °C, for three hours each day for one, two or three weeks. Glial cell activation, neuronal structure, and neuroinflammation were then evaluated by western blotting, immunofluorescence, Nissl staining and co-immunoprecipitation. Microglial activation, accompanied by activation of the NF-κB signaling pathway, release of pro-inflammatory cytokines and loss of Nissl bodies, was observed in mouse hippocampal tissue following cold exposure. We speculate that these phenomena are mediated by the HMGB1/TLR4/NF-κB pathway and closely associated with acetylation of HMGB1 in the hippocampus. These findings provide new insights into the mechanisms of the cold stress response, which should inform the development of new strategies to combat the effects of hypothermia.

To investigate the change of apoptosis protein (Caspase 3) expression when RNA binding motif protein 3 (RBM3) overexpression in swine testicle (ST) cell line under cold stress (32℃) condition.In present study, RBM3 overexpression lentiviral vector (pLenti6/V5-GW/EmGFP-RBM3) and empty viral vector (pLenti6/V5-GW/EmGFP-DEST) with green fluorescent protein (GFP) that were successfully constructed in our laboratory were transfected into ST cells as overexpression virus group (OEV), empty vector virus (EVV) group and establishment of wildtype cell (WTC) group as the control group. The real-time fluorescence quantitative RCR (qPCR) and Western blot were used to detect expression of RBM3 mRNA and protein in each group, then cells in each group were cultured at 37℃ or 32℃ (2 h, 4 h, 8 h), the changes of Caspase 3 expression in each group were detected by enzyme linked immunosorbent assay (ELISA).Relative RBM3 gene and protein expression in OEV group were significantly higher than those in EVV group and WTC group. At the temperature of 37℃ and at 32℃ (2 h, 4 h, 8 h) in cold stress experiment, Caspase 3 expression in OEV group was significantly lower than those in EVV group and WTC group.RBM3 overexpression in ST cell line exposured to cold can significantly decreased the level of Caspase 3 expression, this study provides an experimental basis of RBM3 resistance against apoptosis of ST cells induced by low temperature.

Oreolalax xiangchengensis is an alpine toad species of Megophryidae. We determined its complete mitogenome. The mitogenome has a length of 17,110 bp, containing 13 protein-coding genes, 2 rRNA genes, 23 tRNA genes, and a control region. It has a tandem duplication of tRNA-Met gene and a translocation of tRNA-Trp gene compared with typical vertebrate mitogenomes. Phylogenetic analyses based on the mitogenome sequences of 18 Megophryidae species were conducted to access phylogenetic assignments of O. xiangchengensis.

Cold stress can induce neuronal apoptosis in the hippocampus, but the internal mechanism involving neuronal loss induced by cold stress is not clear. In vivo, male and female C57BL/6 mice were exposed to 4 &deg, C, 3 h per day for 1 week. In vitro, HT22 cells were treated with different concentrations of cortisol (CORT) for 3 h. In vivo, CORT levels in the hippocampus were measured using ELISA, western blotting, and immunohistochemistry to assess the neuronal population and oxidation of the hippocampus. In vitro, western blotting, immunofluorescence, flow cytometry, transmission electron microscopy, and other methods were used to characterize the mechanism of mitochondrial damage induced by CORT. The phenomena of excessive CORT-mediated oxidation stress and neuronal apoptosis were shown in mouse hippocampus tissue following cold exposure, involving mitochondrial oxidative stress and endogenous apoptotic pathway activation. These processes were mediated by acetylation of lysine 9 of histone 3, resulting in upregulation involving Adenosine 5&lsquo, monophosphate (AMP)-activated protein kinase (APMK) phosphorylation and translocation of Nrf2 to the nucleus. In addition, oxidation in male mice was more severe. These findings provide a new understanding of the underlying mechanisms of the cold stress response and explain the apoptosis process induced by CORT, which may influence the selection of animal models in future stress-related studies.

The genusOdorranais widely distributed in the mountains of East and Southeastern Asia. An increasing number of new species in the genus have been recognized especially in the last decade. Phylogenetic studies of theO. schmackerispecies complex with wide distributional range also revealed several cryptic species. Here, we describe a new species in the species complex from Guizhou Province of China. Phylogenetic analyses based on mitochondrial DNA indicated the new species as a monophyly clustered into theOdorranaclade and sister toO. schmackeri, and nuclear DNA also indicated it as an independent lineage separated from its related species. Morphologically, the new species can be distinguished from its congeners based on a combination of the following characters: (1) having smaller body size in males (snout-vent length (SVL)

In this study, we intend to confirm our hypothesis that cold inducible RNA-binding protein (CIRP) can inhibit neuronal apoptosis through suppressing the formation of oxygen free radicals under hypothermia. Primary rat hippocampal neurons were isolated and cultured in vitro, and were divided into five groups: (1) normal control group (37 °C), (2) cells infected by empty viral vector group, (3) CIRP over-expressed group, (4) CIRP knock-down group, and (5) hypothermia control group. Cells in groups 2-5 were cultured under 32 °C, 5% CO2. Apoptosis of hippocampal neurons were detected by Annexin V-FITC/PI staining and flow cytometry; Expression of CIRP was determined by Western blot; Redox-related parameters (T-AOC, GSH-Px, SOD, MDA) were detected by ELISA kits. Results showed that CIRP expression levels were significantly increased (P0.01) and the apoptotic rates were significantly decreased (P0.01) in hypothermia control group and CIRP over-expressed group when compared with normal control group. On the other hand, the apoptotic rate was significantly increased (P0.05) in CIRP knock-down group compared with that in hypothermia control group. The levels of redox parameters in hypothermia control group and CIRP over-expressed group were significantly changed in comparison with those in normal control group, CIRP knock-down group and empty viral vector infected group, respectively (P0.05 or P0.01). These results suggest that up-regulation of CIRP by hypothermia treatment can protect the neuron from apoptosis through suppressing the formation of oxygen free radicals.

We report a feasible method that can precisely control the fission of droplets by modulating the flow resistance using pneumatic valves. Multilayer soft lithography was used to fabricate the valves. They can be used as variable microfluidic resistor (VMR) to dramatically change the flow resistance. A simulation has been done to forecast the behavior of droplets. We used this technique to control break-up of generated droplets and direct their motion. Droplets with different volume ratios were obtained in one chip. To investigate the mechanism, an equivalent electrical circuit was introduced to compare with the fluid network. This method could potentially be applied to different geometries, especially for the microfluidic network consisting of a set of two parallel channels with a common inlet and different outlets in bifurcating channels. Besides, manipulation of bubbles was also demonstrated.

Three Asian green pit vipers were collected in August 2013 during a field trip in Fanjin Mt. National Conservation Area, Guizhou. These specimens were identified as Sinovipera sichuanensis, based on subsequent examination and comparison. This is a new record of the genus Sinovipera and S. sichuanensis in Guizhou, and the first time that male specimens have been collected in the field.

One hundred primers were used to analyze the polymorphism of kinship of Sanjiang White pig and its parents, Landrace and Dongbei Min pig, Yorkshire, French Hybrids (Fh 019 strain), French Yorkshire by RAPD. Amplification results of 15 primers showed obvious polymorphisms, which were analyzed by clustering. The results indicate that the genetic relationship of Sanjiang White Pig, and Landrace, and Dongbei Min pig are respectively closer(d=0.090,,0.175), and that of Sanjiang White pig and Yorkshire (d=0.263), French Yorkshire (d = 0.223), Fh 019 strain (d = 0.580) are respectively further.