Chromosome-level assembly of Gymnocypris eckloni genome.

Gymnocypris eckloni is widely distributed in isolated lakes and the upper reaches of the Yellow River and play significant roles in the trophic web of freshwater communities. In this study, we generated a chromosome-level genome of G. eckloni using PacBio, Illumina and Hi-C sequencing data. The genome consists of 23 pseudo-chromosomes that contain 918.68 Mb of sequence, with a scaffold N50 length of 43.54 Mb. In total, 23,157 genes were annotated, representing 94.80% of the total predicted protein-coding genes. The phylogenetic analysis showed that G. eckloni was most closely related to C. carpio with an estimated divergence time of ~34.8 million years ago. For G. eckloni, we identified a high-quality genome at the chromosome level. This genome will serve as a valuable genomic resource for future research on the evolution and ecology of the schizothoracine fish in the Qinghai-Tibetan Plateau.

Method of Setting Environmental Administrative Fine Amounts.

In China, there are currently different degrees of arbitrariness in setting environmental administrative fines, and in many areas the faults are not equal to the penalties. To construct a more reasonable and feasible environmental punishment strategy where violators are fined in accordance with the severity of their actions, we use mathematical models to determine the specific range of environmental administrative fines based on the idea of realizing the appropriate balance between the interest of the violators and those of the public, meanwhile, law enforcement officers are allowed to use their discretion within a certain range. We use an example to prove that the punishment scheme provided by our models can be used to more effectively supervise violators' illegal behaviors than the penalty clause prescribed by law, and through sensitivity analysis and comparison, we prove that the described models are stable and feasible, and provide advantages over the existing methods. We hope our approach provides intellectual support for maintaining legal order, regulating the environmental administrative fine process, guiding business behaviors, and, most importantly, achieving the goal of protecting the environment.

[Difference in oxygen uptake in skeletal muscles between plateau zokor (Myospalax rufescens baileyi) and plateau pika (Ochotona curzoniac)].

To investigate the difference between the functions of oxygen uptake in skeletal muscle and living habits of plateau zokor (Myospalax rufescens baileyi) and plateau pika (Ochotona curzoniac), the microvessel densities (MVD) of skeletal muscle of plateau zokor, plateau pika and Sprague-Dawley (SD) rat were measured by immunohistochemical staining; the numerical density on area (N(A)) of mitochondria, and surface density (S(V), external surface area density of mitochondria per unit volume of skeletal muscle fiber) were obtained by stereo microscope technique; mRNA levels of myoglobin (Mb) in skeletal muscle were determined by real-time PCR, and the contents of Mb protein in skeletal muscle were determined by spectro-photometer. The results showed that MVD, N(A) and S(V) of mitochondria in skeletal muscle of plateau pika were significantly lower than those of plateau zokor and SD rat (P<0.05). The mRNA levels of Mb gene in skeletal muscle of plateau zokor and plateau pika were notably higher than that of SD rat (P<0.05). There were significant differences in the contents of Mb among these three species, and plateau zokor and SD rat presented the highest and the lowest value, respectively (P<0.05). The results suggest that even though plateau zokor inhabits in the hypoxia environment, most of its skeletal muscle fiber are red muscle fiber. While most of skeletal muscle fibers of plateau pika are white muscle fibers. This kind of white muscle has low MVD, N(A) and S(V) of mitochondria and less content of Mb compared with the red one, suggesting it obtains most energy from aerobic oxidation. The above-mentioned differences in skeletal muscles may be related to not only the different species, but also the different living habits of these two high altitude species.

[Hypoxic adaptation of the hearts of plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae)].

Plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzniae) are native to the Qinghai-Tibet plateau. To study their adaptive mechanisms, the ratios of heart weight to body weight (HW/BW) and right to left ventricular plus septum weights [RV/(LV+S)] were determined; the microvessel density (MVD) of cardiac muscle were measured by immunohistochemical staining; the numerical density on area (N(A)), volume density (V(V)), specific surface (δ), and surface density (S(V)) of mitochondria were obtained by microscopy and stereology; the contents of myoglobin (Mb) and lactic acid (LD), and the activity of lactate dehydrogenase (LDH) in cardiac muscle were analyzed by spectrophotometer. The results showed that the HW/BW of plateau zokor [(4.55±0.26)%] and plateau pika [(4.41±0.38)%] was significantly greater than that of Sprague-Dawley (SD) rat [(3.44±0.41)%] (P<0.05), but the RV/(LV+S) [(22.04±1.98)%, (25.53±3.41)%] was smaller than that of SD rats [(44.23±3.87)%] (P<0.05). The MVD and N(A) of cardiac muscle were 1688.631±250.253 and 0.768±0.123 in SD rat, 2002.888±367.466 and 0.868±0.159 in plateau pika and 2 990.643±389.888 and 1.012±0.133 in plateau zokor. The V(V) of mitochondria in plateau zokor (0.272±0.045) was significantly lower than that in plateau pika (0.343±0.039) and SD rat (0.321±0.048) (P<0.05), while the δ of mitochondria in plateau zokor (9.409±1.238) was higher than that in plateau pika (6.772±0.892) and SD rat (7.287±1.373) (P<0.05). The S(V) of mitochondria in plateau pika (2.322±0.347) was not obviously different from that in plateau zokor (2.468±0.380) and SD rat (2.227±0.377), but that in plateau zokor was significantly higher than that in SD rat (P<0.05). The contents of Mb in cardiac muscle of plateau zokor [(763.33±88.73) nmol/g] and plateau pika [(765.96±28.47) nmol/g] were significantly higher than that of SD rat [(492.38±72.14) nmol/g] (P<0.05), the content of LD in plateau zokor [(0.57±0.06) mmol/L] was obviously higher than that in plateau pika [(0.45±0.06) mmol/L] and SD rat [(0.48±0.02) mmol/L] (P<0.05), and the activity of LDH in plateau zokor [(16.90±2.00) U/mL] and plateau pika [(20.55±2.46) U/mL] were significantly lower than that in SD rat [(38.26±6.78) U/mL] (P<0.05). The percentage of LDH-H in cardiac muscle decreased in order in plateau zokor, plateau pika and SD rat. In conclusion, plateau zokor and plateau pika adapt better to hypoxia than SD rat by increasing the SV of mitochondria, MVD and content of Mb in the cardiac muscle. However, the parameters of mitochondria in the two high-altitude animals are different possibly because of the differences of habitats and habits.

Characterization and phenanthrene sorption of organic matter fractions isolated from organic and mineral soils.

Sorption of phenanthrene (PHE) to humic acid (HA) and humin (HM) fractions isolated from organic and mineral soils was investigated to better understand sorption processes in varying soil types. Samples were characterized by elemental analysis, X-ray photoelectron spectroscopy, 13C nuclear magnetic resonance, and CO2 adsorption. No clear correlation was found between the distribution coefficient (Kd) and the bulk polarity of the soil organic matters (SOMs). By contrast, PHE Kd values generally increased with increasing surface polarity of the tested SOMs, implying that surface polarity may play a more important role in PHE sorption than the bulk one. The organic carbon (OC)-normalized Kd values (Koc) of HMs were higher than those of HAs as a result of the higher aliphatic C contents of HMs. For SOMs isolated from mineral soil (MI-SOMs), part of the aliphatic domains may be tightly associated with minerals and were not accessible to PHE molecules, resulting in lower PHE Koc values of MI-SOMs than the corresponding fractions extracted from the organic soil. This study implies that both chemical characteristics and physical conformation of SOMs are paramount considerations when investigating sorption process of hydrophobic organic compounds in soils.

Studies of the cryopreservation condition of Gymnocypris przewalskii spermatozoa.

The endemic naked carp (Gymnocypris przewalskii Kessler, 1876) plays an important role in the maintenance of the distinctive ecological system of Lake Qinghai at 3.2km altitude on the Qinghai-Tibet Plateau in China. This study aimed to develop a cryopreservation protocol for Gymnocypris przewalskii spermatozoa. Semen was collected from mature individuals during migration and frozen using the liquid nitrogen vapor method. The influence of different cryoprotectants and three extenders on the post-thaw quality of the sperm was analyzed. The highest sperm motility rate and longest motility time after cryopreservation were achieved by combining Ringer's solution with 15% ethylene glycol (P<0.05). The fertilization rate of this cryopreserved semen was 15.26±4.54%. This study thus provides a valuable method for the cryopreservation of the sperm of this important endangered fish species.

Repeated administration of pioglitazone attenuates development of hyperalgesia in a rat model of neuropathic pain.

Recent studies indicate the central neuroimmune and neuroinflammation activation play a critical role in the pathological states of pain. Pioglitazone, a potent synthetic agonists of PPARgamma, has shown to control neuroinflammation in many nervous system-related disorders. The present study was designed to explore the effects of pioglitazone in treating neuropathic pain and its possible neuroimmune mechanisms in the neuropathic pain using lumbar 5 (L5) spinal nerve transection rat model. L5 spinal nerve transection was done to produce hyperalgesia in rats. Pioglitazone (2.5, 5, and 10 mg/kg) was orally administered daily for 14 days, beginning from 1 hour before nerve transection. Mechanical hyperalgesia was measured using Von-Frey filament tests before and after the surgery. Rats were then sacrificed on day 14 postsurgery. The mRNA of inflammatory cytokines such as tumor necrosis factor (TNF-alpha), interleukin (IL-1beta) and nuclear factor kappa B (NF-kappaB) activity in brain were detected using reverse transcription-polymerase chain reaction and electrophoretic mobility shift assay. We found that pioglitazone (5 and 10 mg/kg) can markedly attenuate mechanical hyperalgesia produced by nerve transection, most significantly on the 14th day. The elevated TNF-alpha, IL-1beta, and NF-kappaB in brain were accordingly reduced. Our data could conclude that pioglitazone has ameliorative potential in attenuating the painful state associated with L5 nerve transection, which may further be attributed to inhibiting cerebral proinflammatory cytokines production and NF-kappaB activation in central nervous system.