Preparation of the monoclonal antibody against Nile tilapia Igλ and study on the Igλ+ B cell subset in Nile tilapia.

Immunoglobulins (Igs) are important effector molecules that mediate humoral immunity. A typical Ig consists of two heavy and two light chains. In teleosts, three Ig heavy chain isotypes (Igµ, Igδ and Igτ) and three Ig light chain isotypes (Igκ, Igλ and Igσ) have been identified. Compared to the heavy chains, teleost Ig light chains have been poorly studied due to the lack of antibodies. In this study, a mouse anti-Nile tilapia Igλ monoclonal antibody (mAb) was prepared, which could specifically recognize Igλ in serum and Igλ+ B cells in tissues. Further, the composition of IgM+ and Igλ+ B cell subsets was analyzed using this antibody and a mouse anti-tilapia IgM heavy chain mAb. The ratio of IgM+Igλ+ B cells to total IgM+ B cells in head kidney and peripheral blood was about 30%, while that in spleen was about 50%; the ratio of IgM-Igλ+ B cells to total Igλ+ B cells in head kidney and peripheral blood was about 45%, while that in spleen was about 25%. The IgM-Igλ+ B cells was speculated to be IgT+ B cells. Finally, we detected an increase in the level of specific antibodies against the surface antigen-Sip of Streptococcus agalactiae in serum after S. agalactiae infection, indicating that mouse anti-tilapia Igλ mAb can be used to detect the antibody level after immunization of Nile tilapia, which lays a foundation for the evaluation of immunization effect of tilapia vaccine.

Design, synthesis and bioactivities evaluation of novel oleanolic acid derivatives as potent PI3K inhibitors.

Oleanolic acid has previously been shown to possess PI3K inhibitory activity, thus, the purpose of this work was to generate a series of derivatives that improve the potency. Twenty rationally designed oleanolic acid derivatives were synthesized and tested the cytotoxicity and PI3K inhibitory activity. The results suggested that attachment of additional structural elements such as association of thiazole group to A ring and insertion of phenylurea group was important for increasing activities. The most active derivative was compound II2, which exhibited PI3K inhibitory activity (IC50 = 58.42 nmol/l) and improved interaction with activity site of PI3K according with docking studies.

Iron-Catalyzed Oxidative Amination of Benzylic C(sp3)-H Bonds with Anilines.

Iron-catalyzed oxidative amination of benzylic C(sp3)-H bonds with anilines bearing electron-withdrawing groups (EWGs) or electron-donating groups (EDGs) is realized based on simple variations of N-substituents on imidazolium cations in novel ionic Fe(III) complexes. The structural modification of the imidazolium cation resulted in regulation of the redox potential and the catalytic performance of the iron metal center. Using DTBP as oxidant, [HItBu][FeBr4] showed the highest catalytic activity for anilines bearing EWGs, while [HIPym][FeBr4] was more efficient for EDG-substituted anilines. This work provides alternative access to benzylamines with the advantages of both a wide substrate scope and iron catalysis.

Molecular Characterization and Expression Analysis of Intercellular Adhesion Molecule-1 (ICAM-1) Genes in Rainbow Trout (Oncorhynchus mykiss) in Response to Viral, Bacterial and Parasitic Challenge.

The intercellular adhesion molecule-1 (ICAM-1), known as CD54, is a transmembrane cell surface glycoprotein that interacts with two integrins (i.e., LFA-1 and Mac-l) important for trans-endothelial migration of leukocytes. The level of ICAM-1 expression is upregulated in response to some inflammatory stimulations, including pathogen infection and proinflammatory cytokines. Yet, to date, our knowledge regarding the functional role of ICAM-1 in teleost fish remains largely unknown. In this study, we cloned and characterized the sequence of ICAM-1 in rainbow trout (Oncorhynchus mykiss) for the first time, which exhibited that the molecular features of ICAM-1 in fishes were relatively conserved compared with human ICAM-1. The transcriptional level of ICAM-1 was detected in 12 different tissues, and we found high expression of this gene in the head kidney, spleen, gills, skin, nose, and pharynx. Moreover, upon stimulation with infectious hematopoietic necrosis virus (IHNV), Flavobacterium columnare G4 (F. columnare), and Ichthyophthirius multifiliis (Ich) in rainbow trout, the morphological changes were observed in the skin and gills, and enhanced expression of ICAM-1 mRNA was detected both in the systemic and mucosal tissues. These results indicate that ICAM-1 may be implicated in the mucosal immune responses to viral, bacterial, and parasitic infections in teleost fish, meaning that ICAM-1 emerges as a master regulator of mucosal immune responses against pathogen infections in teleost fish.

Design and synthesis of VEGFR-2 inhibitors based on oleanolic acid moiety.

In this study, twenty-four oleanolic acid (OA) derivatives were rationally designed based on molecule docking studies and their VEGFR-2 inhibitory activities were tested by Homogeneous time-resolved fluorescence (HTRF) method in vitro. All of the synthesized compounds were identified as new compounds, and the structures of these compounds were determined by 1H-NMR and ESI-MS. In the screening for VEGFR-2 inhibitors, compounds I6 and I7 exhibited excellent inhibitory effect. The results indicated that insertion of phenylurea group with a linker at position C-28 of OA can increase the activity against VEGFR-2 significantly. [Formula: see text].

Different elevational patterns of rodent species richness between the southern and northern slopes of a mountain.

Studies on elevational gradients in biodiversity have accumulated in recent decades. However, few studies have compared the elevational patterns of diversity between the different slopes of a single mountain. We investigated the elevational distribution of rodent diversity (alpha and beta diversity) and its underlying mechanisms along the southern and northern slopes of Mt. Taibai, the highest mountain in the Qinling Mountains, China. The species richness of rodents on the two slopes showed distinct distribution patterns, with a monotonically decreasing pattern found along the southern slope and a hump-shaped elevational pattern evident along the northern slope. Multi-model inference suggested that temperature was an important explanatory factor for the richness pattern along the southern slope, and the mid-domain effect (MDE) was important in explaining the richness pattern along the northern slope. The two slopes also greatly differed in the elevational patterns of species turnover, with the southern slope demonstrating a U-shaped curve and the northern slope possessing a roughly hump-shaped pattern. Our results suggest that even within the same mountain, organisms inhabiting different slopes may possess distinct diversity patterns, and the underlying mechanisms may also differ. The potential role of the factors associated with slope aspect in shaping diversity, therefore, cannot be ignored.

Phylogeographical structure and demographic expansion in the endemic alpine stream salamander (Hynobiidae: Batrachuperus) of the Qinling Mountains.

The Qinling Mountains of China provide an excellent study area for assessing the effect of Pleistocene climatic oscillations and paleogeological events on intraspecific diversification. To assess genetic diversity of an endemic stream salamander, Batrachuperus tibetanus, for its conservation, a phylogeographical survey was performed based on mitochondrial DNA and morphological data. The mitochondrial data revealed three lineages of B. tibetanus in the Qinling Mountains. A lineage present in the northwestern Qinling Mountains groups with the Tibet lineage of B. tibetanus, and the remaining Qinling populations are eastern and western lineages that separated ~3-4 million years ago (Ma). The eastern and western Qinling lineage delineation is supported by three morphological variables (snout length, eye diameter and axilla-groin length). The divergence of the two major lineages was likely caused by orogenesis of the Qinling Mountains during the late Cenozoic, and the two lineages were subsequently affected at different levels by Pleistocene climatic oscillations showing different signals of demographic expansion. A large suitable area of B. tibetanus through the Qinling Mountains since the last glacial maximum (LGM) indicated the adaptation of this species to the climatic changes. However, low genetic diversity within populations indicate the urgency of preserving the vulnerable populations and endemic lineages.

Effects of cattle grazing on small mammal communities in the Hulunber meadow steppe.

Small mammals play important roles in many ecosystems, and understanding their response to disturbances such as cattle grazing is fundamental for developing sustainable land use strategies. However, how small mammals respond to cattle grazing remains controversial. A potential cause is that most of previous studies adopt rather simple experimental designs based solely on the presence/absence of grazing, and are thus unable to detect any complex relationships between diversity and grazing intensity. In this study, we conducted manipulated experiments in the Hulunber meadow steppe to survey small mammal community structures under four levels of grazing intensities. We found dramatic changes in species composition in native small mammal communities when grazing intensity reached intermediate levels (0.46 animal unit/ha). As grazing intensity increased, Spermophilus dauricus gradually became the single dominant species. Species richness and diversity of small mammals in ungrazed and lightly grazed (0.23 animal unit/ha) area were much higher than in intermediately and heavily grazed area. We did not detect a humped relationship between small mammal diversity and disturbance levels predicted by the intermediate disturbance hypothesis (IDH). Our study highlighted the necessity of conducting manipulated experiments under multiple grazing intensities.

Role of New Nature Reserve in Assisting Endangered Species Conservation - Case Study of Giant Pandas in the Northern Qionglai Mountains, China.

The creation of nature reserves is the most direct way to save endangered species populations and their habitat. Development of the giant panda (Ailuropoda melanoleuca) nature reserve network in China was initiated in the 1960s, though the effort to create new reserves boomed considerably after the year 2000. Given this rapid development of protected areas in panda habitats, and the potential conflicting interests between conservation administrations and local economic development, it is essential to assess the role of new nature reserves in the overall giant panda conservation effort and reserve network. We utilized data from national giant panda surveys conducted in 2000 and 2012 to compare the size, spatial use, and distribution of panda populations, as well as the habitat suitability and connectivity in the Northern Qionglai Mountains between the two survey years. Our results show that although the total giant panda population in the study area did not change remarkably, local changes did occur. Most notably, the population in Wolong Nature Reserve declined by 27.3% (N = 39) and the population in Caopo Nature Reserve increased by 71.4% (N = 29) over the 12-year study period. We also found habitat suitability and availability decreased in both Wolong (12.4%) and Caopo (7.4%), but that the relative density of giant pandas declined (19.2%) and increased (84.6%) at each site, respectively. The distance between centers of high IUA were more distant in 2012 (14.1±1.9km) than that in 2000 (6.1±0.9km; t = -7.4, df = 5, p = 0.001), showing a scattered spatial pattern. Habitat availability decreased by 42% within the corridor between the two reserves, however panda occurrences in the corridor increased 24.6%. Compared to the total number of encounters, the proportion of the corridor increased 45.76%. Our results show the importance and success of the newly established Caopo to the conservation of giant pandas, and how crucial it is to identify and repair reserve corridors. Furthermore, we propose criteria for future nature reserve network management and investment, which is applicable for other endangered species conservation practices.

Effects of Different Anesthetic Methods on Cellular Immune and Neuroendocrine Functions in Patients With Hepatocellular Carcinoma Before and After Surgery.

BACKGROUND: Many anesthesia methods have been studies in hepatocellular carcinoma (HCC). We aimed to explore the effects of combined intravenous and inhalation anesthesia and combined general and epidural anesthesia on cellular immune function and neuroendocrine function in patients with HCC before and after surgery. METHODS: Between September 2012 and April 2014, 72 patients who underwent a hepatectomy in our hospital were enrolled. RESULTS: Compared with the combined intravenous and inhalation anesthesia group, the combined general and epidural anesthesia group demonstrated increased CD4+ /CD8+ T cells 0 hr after surgery, increased CD3+ , CD4+ , CD4+ /CD8+ cells, and IFN-γ levels 12 hr after surgery, and increased CD3+ , CD4+ , and CD4+ /CD8+ cells 24 hr after surgery (all P < 0.05). At 72 hr after surgery, the levels of ACTH and Cor in the combined general and epidural anesthesia group, and the levels of CD3+ , CD4+ , CD4+ /CD8+ cells, and IFN-γ in both the combined intravenous and inhalation anesthesia and the combined general and epidural anesthesia groups decreased to pre-surgery levels. Significant differences were observed in the comparisons of CD3+ , IL-6, and IL-10 between the combined intravenous and inhalation anesthesia and the combined general and epidural anesthesia groups 72 hr after surgery (all P < 0.05). CONCLUSION: Our results revealed that combined general and epidural anesthesia plays a crucial role in hepatectomy via the mitigation of the inhibition of immunologic function in HCC patients during the perioperative period. Combined general and epidural anesthesia also hastens the recovery of immunologic suppression after surgery, which can provide a certain reference for the selection of clinical anesthesia in the treatment of HCC.

[Advances in study of Survivin inhibitors].

Survivin, a member of the inhibitor of apoptosis proteins family, is considered to be an important target of anticancer treatment for its key role in the control of cell division and cell apoptosis. Currently, only a few Survivin inhibitors have been developed, and most of them reduce Survivin level by interacting with other biomolecules instead of directly interacting with Survivin protein. This review summarizes the structure of Survivin protein, the mechanism of action and research progress of Survivin inhibitors, which may has a great significance in the study of selective Survivin inhibitors in the future.

Spectroscopic investigation on the sonodynamic damage to protein in the presence of eosine B.

In this work, bovine serum albumin (BSA) and eosine B (EB) were selected as a model protein and sonosensitizer, respectively. The sonodynamic damage to protein in the presence of EB and its mechanism were studied by means of absorption and fluorescence spectra. The results indicated that the synergistic effects of ultrasound and EB can efficiently damage the BSA molecules, and the damage of protein could be mainly due to the generation of reactive oxygen species (ROS). The damage degree of protein increased with the increase of ultrasonic time and EB concentration because of the increased quantities of ROS. Hydroxyl free radical (OH) was the major mediators of the ultrasound-inducing proteins damage in the presence of EB. In addition, the quantities of ROS from the diphenylcarbazide (DPCI)-EB solutions and the DPCI-fluorescein (FS) solutions with or without ROS scavengers were contrasted, respectively. The results show that FS mainly produce OH, but the quantities of ROS in the presence of FS were lower than those of EB, which indicates that the nitro and bromine substituent groups on the benzene ring of FS increase the quantity of ROS, but do not change the kinds of ROS.

Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity.

Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher's attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56±7.3nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

Habitat selection by Eld's deer following relocation to a patchy landscape.

An emerging issue in wildlife conservation is the re-establishment of viable populations of endangered species in suitable habitats. Here, we studied habitat selection by a population of Hainan Eld's deer (Cervus eldi) relocated to a patchy landscape of farmland and forest. Hainan Eld's deer were pushed to the brink of extinction in the 1970s, but their population expanded rapidly from 26 to more than 1000 individuals by 2003 through effective reserve protection. As part of a wider relocation and population management strategy, 131 deer were removed from the reserve and reintroduced into a farmland-forest landscape in 2005. Habitat use under a context of human disturbance was surveyed by monitoring 19 radio-collared animals. The majority of deer locations (77%) were within 0.6-2 km of villages. Annual home ranges of these collared deer averaged 725 ha (SD 436), which was 55% of the size of the reserve from which they had originated. The annual home ranges contained 54% shrub-grassland, 26% forest and 15% farmland. The relocated deer population selected landscape comprising slash-and-burn agriculture and forest, and avoided both intensively farmed areas and areas containing only forest. Within the selected landscape, deer preferred swiddens and shrub-grasslands. Forests above 300 m in elevation were avoided, whereas forests below 300 m in elevation were overrepresented during the dry season and randomly used during the wet season. Our findings show that reintroduced deer can utilize disturbed habitats, and further demonstrate that subsistence agroforest ecosystems have the capacity to sustain endangered ungulates.

Passive movement improves the learning and memory function of rats with cerebral infarction by inhibiting neuron cell apoptosis.

Passive movement has been found to improve evidently ischemic stroke patients' impaired capacity of learning and memory, but the optimal time window of initiating the therapy and the underlying mechanism are not fully understood. In this study, the effect of passive movement at different time windows on learning and memory of rats with cerebral infarction was detected. The results showed that the expression of caspase-3 and escape latency in the passive movement group were all considerably lower than those in the model group (P < 0.05), while the expression of Bcl-2 mRNA was significantly higher than those in the model group (P < 0.05). Moreover, we found that there were most significant changes of escape latency and expressions of Bcl-2 mRNA and caspase-3 when the therapy started at 24 h after focal cerebral infarction. These results suggest that passive movement is able to contribute to the recovery of learning and memory of rats with cerebral infarction, which is partially mediated by inhibiting neuron cell apoptosis, and the optimal therapeutic time is at 24 h after cerebral infarction.

Synthesis and antitumor activity evaluation of new asiatic acid derivatives.

Twelve novel asiatic acid (AA) derivatives were designed and synthesized. Their structures were confirmed using NMR, MS, and IR spectra. Their in vitro cytotoxicities on various cancer cell lines (HeLa, HepG2, BGC-823, and SKOV3) were evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Most of the derivatives were found to have stronger cell growth inhibitory activity than AA. Among them, compounds 5-8 and 11 with substituted amide group at C-28 exhibited more potent cytotoxicity than AA, Gefitinib, and etoposide (positive control).

Effect of HbA1c combined FPG on screening diabetes in health check-up.

OBJECTIVE: To appraise the effectiveness of HbA(1c) and fasting plasma glucose (FPG) on screening diabetes in health check-up. METHODS: A total of 1,337 individuals (male 850, female 487), aged 27 to 91 years with HbA(1c) test were included. Participates with HbA(1c) ≥6.0% or FPG≥6.1 mmol/L underwent oral glucose tolerance test (OGTT). Diabetes mellitus was diagnosed according to the criteria of WHO in 1999, FPG≥7.0 mmol/L and/or OGTT 2 h-postload plasm glucose (2 h-PG)≥11.1 mmol/L. The sensitivity and specificity of HbA(1c) thresholds and FPG or combination test on screening of diabetes were analyzed. RESULTS: A total of 842 subjects had HbA(1c) <6.0%, in which 32 had isolated FPG≥6.1 mmol/L, of 495 had HbA(1c)≥6.0%. Subjects with HbA(1c)≥6.0% had significant increased disorder indexes than those with HbA(1c)<6.0%. 527 subjects who had HbA(1c)≥6.0% or FPG≥6.1 mmol/L underwent OGTT. A total of 234 subjects were newly diagnosed diabetes, including 123 (123/234, 52.56%) with FPG≥7.0 mmol/L, and 111 subjects (111/234, 47.43%) with isolated 2 h-PG≥11.1 mmol/L. Among 234 new diabetes, 91.88% (215 subjects) had HbA(1c)≥6.3%, and 77.40% (181 subjects) had HbA(1c)≥6.5%. HbA(1c)≥6.3% combined FPG ≥7.0 mmol/L increased the positive rate of newly diagnosed diabetes from 91.88% to 96.58%. CONCLUSIONS: HbA(1c) is a practical and convenient tool for screening undiagnosed diabetes in routine health check-up of a large population. Combined use of HbA(1c)≥6.3% and/or FPG≥7.0 mmol/L is efficient for early detection of diabetes.

Does a temperate ungulate that breeds in summer exhibit rut-induced hypophagia? Analysis of time budgets of male takin (Budorcas taxicolor) in Sichuan, China.

Mammals maximize fitness by optimizing time and energy allocation between reproduction and survival. Describing time budgets is a way to understand a species' constraints in energy allocation. We describe a time budget for male takin (Budorcas taxicolor) in Tangjiahe Nature Reserve, China, to better understand rut-induced hypophagia, which is frequently observed in temperate ungulates that breed in autumn or in winter. Observations generally occurred at two elevations (1200-1600m and 2600-3200m), using 20-min focal animal scan sampling from 2007 to 2009. Feeding behaviors accounted for the majority in takin's time budget (61.1%) during daylight hours, relative to the other observed behaviors, such as rest (14.1%), alert behavior (10.2%) and locomotion (6.8%). We found a negative correlation between feeding behavior and rutting behavior during the rutting season. A ratio of feeding time to resting time increased from pre-rut to rut, while resting behavior did not change significantly across seasons. These results suggest the "energy saving" hypothesis could explain reduced foraging in male takin during the rut, but aspects of the species biology suggest that hypotheses for rut-induced hypophagia developed for other temperate ungulates do not apply to takin. We suggest that the unusual summer rutting season of takin releases males from the energy constraints encountered by temperate ungulates that breed in the autumn and has other benefits for offspring survival. Further research should be conducted on ungulates that exhibit rut during the summer and tropical ungulates that might not experience limited food availability following the mating season to improve our understanding on rut-induced hypophagia.

Foraging behavior of the midday gerbil (Meriones meridianus): combined effects of distance and microhabitat.

We used the giving-up density (GUD) method and direct observation to study the combined effects of travel distance and microhabitat on foraging behavior of the midday gerbil (Meriones meridianus), which often acts as a central place forager. We provided animals with artificial seed trays in which dry and unhusked pumpkin seeds were mixed with fine sand. Gerbils practiced an eat-and-carry strategy in patches of bush microhabitat that were far from central places (BF patches), and tended to carry all seeds back in the other three treatments. Resource protection, predation risk avoidance and the balance between future and present value of food items may contribute to the eat-and-carry strategy. When distance was held constant, GUDs in open patches were higher than in bush patches, which was consistent with most studies. When microhabitat was held constant, GUDs in nearer patches were normally lower than in farther patches. In most cases, gerbils preferred to carry more seeds back rather than consume them immediately. We concluded that this tendency was due to the gerbils balancing the factors of future value and present value, and individual fitness and inclusive fitness.