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This study employs stable isotope analysis to investigate the mechanisms of cadmium (Cd) and zinc (Zn) interaction in the metal hyperaccumulating plant species Sedum plumbizincicola. To this end, the Cd and Zn isotope compositions of root, stem, leaf, and xylem sap samples were determined during metal uptake and translocation at different Cd and Zn concentrations. The enrichment of light isotopes of both elements in plants during uptake was less pronounced at low metal supply levels, likely reflecting the switch from a low-affinity to a high-affinity transport system at lower levels of external metal supply. The lower δ114/110Cd values of xylem sap when treated with a metabolic inhibitor decreasing the active Cd uptake further supports the preference of heavier Cd isotopes during high-affinity transport. The Δ66Znplant-initial solution or Δ66Znplant-final solution values were similar at different Cd concentrations, indicating negligible interaction of Cd in the Zn uptake process. However, decreasing Zn supply levels significantly increased the enrichment of light Cd isotopes in plants (Δ114/110Cd = -0.08‰) in low-Cd treatments but reduced the enrichment of light Cd isotopes in plants (Δ114/110Cd = 0.08‰) under high Cd conditions. A systematic enrichment of heavy Cd and light Zn isotopes was found in root-to-shoot translocation of the metals. The Cd concentrations of the growth solutions thereby had no significant impact on Zn isotope fractionation during root-to-shoot translocation. However, the Δ114/110Cdtranslocation values hint at possible competition between Cd and Zn for transporters during root-to-shoot transfer and this may impact the transport pathway of Cd. The stable isotope data demonstrate that the interactions between the two metals influenced the uptake and transport mechanisms of Cd in S. plumbizincicola but had little effect on those of Zn.

Green plants provide food, energy and oxygen sources for human beings and animals on Earth through photosynthesis, which is essential to maintain regional ecological balance. However, few studies have focused on the natural oxygen supply capacity of urban green spaces. As a companion to photosynthesis in leaves, solar-induced chlorophyll fluorescence (SIF) contains abundant photosynthetic information. Currently, satellite-based SIF observations are considered to be a rapid and nondestructive ‘indicator’ of plant photosynthesis, which provides an alternative way to quantitatively assess the spatio-temporal dynamics of oxygen supply capacity in urban green spaces. This study examined the spatial patterns, long-term trends, and environmental control factors of SIF in the nine central cities in China from 2001 to 2020 based on the time-series of the global reconstructed GOSIF-v2 SIF dataset. The results were as follows: (1) There was a contrasting spatial difference between southern and northern cities in China, and multi-year mean SIF values of the southern cities were generally higher than those of the northern cities; (2) The interannual dynamics of SIF in each city generally showed an upward trend, with fluctuations, and the intraannual seasonal differences were more significant in northern cities than those in the southern cities; (3) The spatial trend analysis showed that Beijing, Guangzhou, and Chongqing have had the most significant improvements, followed by Xi’an, Wuhan, Chengdu, and Zhengzhou, while Tianjin and Shanghai have had the least improvements; and (4) The expansion of construction land has exerted significant impacts on the dynamics of the SIF trend in several cities, but it is not the only factor. All analyses indicated that the improvement of vegetation structure and function in the area can offset its negative effect.

We established a model based on the “Malmquist index” to evaluate the dynamic performance of scientific and technological innovation in seven Chinese universities, which included five categories: changes in pure technical efficiency, changes in production technology, changes in scale efficiency, changes in organizational management performance, and changes in comprehensive efficiency. Seven domestic first-class universities were studied, and empirical analysis was performed based on data related to their scientific and technological innovation during three time periods (2013-2014, 2014-2015, and 2015-2016). Our results demonstrated that the contribution rate for each index was different for each university across the three time periods, and that Chinese universities is gradually transitioning from resource allocation to technical efficiency. In this paper, we explored the transformation from effect to efficiency and static to dynamic in the evaluation of the scientific and technological output of colleges and universities, and constructed systematic and unique multi-evaluation system for the scientific and technological output of universities.

As a main desert plant from arid regions of Central Asia, Populus euphratica always encounters with nitrogen shortage in its long life, apart from salt or drought stress. However, it remains unknown how this species responds to low nitrogen and combined stresses of low nitrogen and salinity. Thus, saplings of P. euphratica with uniform size were exposed to normal or low nitrogen condition (150 and 15 ppm ammonium nitrate separately) individually or in combination with salinity. Under low nitrogen conditions we found a positive effect on P. euphratica root growth, which could be associated to high level of nitrogen allocation to support root growth and effective regulation of nitrogen assimilation in comparison with the other poplar species reported before. Under salt stress the root growth of P. euphratica was significantly inhibited, with the side effects of oxidative stress, as saplings stored higher Na

Background: Interleukin-33 (IL-33) has been implicated in nociceptive pain behaviors. However, the underlying molecular and cellular mechanisms remain unclear. Methods: Using electrophysiological recording, immunoblot analysis, immunofluorescence labeling, reverse transcription-PCR, siRNA-mediated knockdown approach and behavior tests, we determined the role of IL-33 in regulating sensory neuronal excitability and pain sensitivity mediated by A-type K+ channels. Results: IL-33 decreased A-type transient outward K+ currents (IA) in small-sized DRG neurons in a concentration-dependent manner, whereas the delayed rectifier currents (IDR) remained unaffected. This IL-33-induced IA decrease was dependent on suppression of the tumorigenicity 2 (ST2) receptor and was associated with a hyperpolarizing shift in the steady-state inactivation. Antagonism of Syk abrogated the IL-33-induced IA response, while inhibition of JAK2 and PKA elicited no such effect. Exposure of DRG cells to IL-33 increased the activity of Akt, but surprisingly, neither Akt nor PI3K influenced the IL-33-induced IA response. IL-33 increased the level of phosphorylated p38 mitogen-activated protein kinase (MAPK). Chemical inhibition of p38 and genetic siRNA knockdown of p38 beta (p38β), but not p38α, abrogated the IA response induced by IL-33. Moreover, IL-33 increased neuronal excitability of DRG neurons and facilitated peripheral pain sensitivity in mice; both of these effects were occluded by IA blockade. Conclusions: Our present study reveals a novel mechanism by which IL-33/ST2 suppresses IA via a Syk-dependent p38β signaling pathway. This mechanism thereby increases DRG neuronal excitability and pain sensitivity in mice. Targeting IL-33/ST2-mediated p38β signaling may represent a therapeutic approach to ameliorate pain behaviors.

microRNA (miRNA)–mediated gene regulation has been studied as a therapeutic approach, but its functional regulatory mechanism in neuropathic pain is not well understood. Here, we identify that miRNA-32-5p (miR-32-5p) is a functional RNA in regulating trigeminal-mediated neuropathic pain. High-throughput sequencing and qPCR analysis showed that miR-32-5p was the most down-regulated miRNA in the injured trigeminal ganglion (TG) of rats. Intra-TG injection of miR-32-5p agomir or overexpression of miR-32-5p by lentiviral delivery in neurons of the injured TG attenuated established trigeminal neuropathic pain. miR-32-5p overexpression did not affect acute physiological pain, while miR-32-5p down-regulation in intact rats was sufficient to cause pain-related behaviors. Nerve injury increased the methylated histone occupancy of binding sites for the transcription factor glucocorticoid receptor in the miR-32-5p promoter region. Inhibition of the enzymes that catalyze H3K9me2 and H3K27me3 restored the expression of miR-32-5p and markedly attenuated pain behaviors. Further, miR-32-5p–targeted Cav3.2 T-type Ca2+ channels and decreased miR-32-5p associated with neuropathic pain caused an increase in Cav3.2 protein expression and T-type channel currents. Conversely, miR-32-5p overexpression in injured TG suppressed the increased expression of Cav3.2 and reversed mechanical allodynia. Together, we conclude that histone methylation-mediated miR-32-5p down-regulation in TG neurons regulates trigeminal neuropathic pain by targeting Cav3.2 channels.

Soil salinity is a widespread stress in semi-arid forests worldwide, but how to manage nitrogen (N) nutrition to improve plant saline tolerance remains unclear. Here, the cuttings of a widely distributed poplar from central Asia, Populus russikki Jabl., were exposed to either normal or low nitrogen (LN) concentrations for two weeks in semi-controlled greenhouse, and then they were added with moderate salt solution or not for another two weeks to evaluate their physiological, biochemical, metabolites and transcriptomic profile changes. LN-pretreating alleviated the toxicity caused by the subsequent salt stress in the poplar plants, demonstrated by a significant reduction in the influx of Na+ and Cl- and improvement of the K+/Na+ ratio. The other salt-stressed traits were also ameliarated, indicated by the variations of chlorophyll content, PSII photochemical activity and lipid peroxidation. Stress alleviation resulted from two different processes. First, LN pretreatment caused a significant increase of non-structural carbohydrates (NSC), allowed for an increased production of osmolytes and a higher potential fueling ion transport under subsequent salt condition, along with increased transcript levels of the cation/H+ ATPase. Second, LN pretreatment enhanced the transcript levels of stress signaling components and phytohormones pathway as well as antioxidant enzyme activities. The results indicate that early restrictions of N supply could enhance posterior survival under saline stress in poplar plants, which is important for plantation programs and restoration activities in semi-arid areas. This research was supported by Natural Science Foundation of China ( 31770644 and 31270660 ), Project of Innovation research team in Sichuan Education Administration in China (No. 13TD0023 ).

The emerging Non-Volatile Memory (NVM) attracts attentions in recent years because of its fast reading/writing speed, high density, low idle power and persistence. With such characteristics, NVM can be applied to application-specific embedded systems to improve performance such as automotive systems. However, NVM has limitation on writing operations and is consequently not suitable to be applied in writing sensitive systems. In this paper, we propose a Vehicle Data Management (VDM) approach based on a hybrid DRAM and NVM architecture for automotive systems. VDM approach includes automotive dynamic data management, vehicle-specific wear-levelling and vehicle fault tolerance. Automotive dynamic data management can distinguish the hot or cold data from vehicle Electronic Control Units (ECUs) and sensors, and then manage the data to reduce the writing operations on NVM for the hybrid main memory. Vehicle-specific wear-levelling can balance the wear to prolong the lifetime of NVM. A fault tolerance mechanism is also provided for NVM to recover the fault blocks. The experimental results show that both automotive dynamic data management and vehicle-specific wear-levelling can effectively reduce the writing operations compared with other approaches. Vehicle fault tolerance can achieve a high recovery rate and a low time overhead when error rate is not very high.

Background Cholecystokinin (CCK) is implicated in the regulation of nociceptive sensitivity of primary afferent neurons. Nevertheless, the underlying cellular and molecular mechanisms remain unknown. Methods Using patch clamp recording, western blot analysis, immunofluorescent labelling, enzyme-linked immunosorbent assays, adenovirus-mediated shRNA knockdown and animal behaviour tests, we studied the effects of CCK-8 on the sensory neuronal excitability and peripheral pain sensitivity mediated by A-type K+ channels. Results CCK-8 reversibly and concentration-dependently decreased A-type K+ channel (I A) in small-sized dorsal root ganglion (DRG) neurons through the activation of CCK type B receptor (CCK-BR), while the sustained delayed rectifier K+ current was unaffected. The intracellular subunit of CCK-BR coimmunoprecipitated with Gαo. Blocking G-protein signaling with pertussis toxin or by the intracellular application of anti-Gβ antibody reversed the inhibitory effects of CCK-8. Antagonism of phosphatidylinositol 3-kinase (PI3K) but not of its common downstream target Akts abolished the CCK-BR-mediated I A response. CCK-8 application significantly activated JNK mitogen-activated protein kinase. Antagonism of either JNK or c-Src prevented the CCK-BR-mediated I A decrease, whereas c-Src inhibition attenuated the CCK-8-induced p-JNK activation. Application of CCK-8 enhanced the action potential firing rate of DRG neurons and elicited mechanical and thermal pain hypersensitivity in mice. These effects were mediated by CCK-BR and were occluded by I A blockade. Conclusion Our findings indicate that CCK-8 attenuated I A through CCK-BR that is coupled to the Gβγ-dependent PI3K and c-Src-mediated JNK pathways, thereby enhancing the sensory neuronal excitability in DRG neurons and peripheral pain sensitivity in mice.

alpha-Melanocyte-stimulating hormone (-MSH) has been shown to be involved in nociception, but the underlying molecular mechanisms remain largely unknown. In this study, we report that -MSH suppresses the transient outward A-type K+ current (I-A) in trigeminal ganglion (TG) neurons and thereby modulates neuronal excitability and peripheral pain sensitivity in rats. Exposing small-diameter TG neurons to -MSH concentration-dependently decreased I-A. This -MSH-induced I-A decrease was dependent on the melanocortin type 4 receptor (MC4R) and associated with a hyperpolarizing shift in the voltage dependence of A-type K+ channel inactivation. Chemical inhibition of phosphatidylinositol 3-kinase (PI3K) with wortmannin or of class I PI3Ks with the selective inhibitor CH5132799 prevented the MC4R-mediated I-A response. Blocking G(i/o)-protein signaling with pertussis toxin or by dialysis of TG neurons with the G-blocking synthetic peptide QEHA abolished the -MSH-mediated decrease in I-A. Further, -MSH increased the expression levels of phospho-p38 mitogen-activated protein kinase, and pharmacological or genetic inhibition of p38 abrogated the -MSH-induced I-A response. Additionally, -MSH significantly increased the action potential firing rate of TG neurons and increased the sensitivity of rats to mechanical stimuli applied to the buccal pad area, and both effects were abrogated by I-A blockade. Taken together, our findings suggest that -MSH suppresses I-A by activating MC4R, which is coupled sequentially to the G complex of the G(i/o)-protein and downstream class I PI3K-dependent p38 signaling, thereby increasing TG neuronal excitability and mechanical pain sensitivity in rats.

Cotton fabric was modified with Na2EDTA using a hydrothermal method to introduce surface hydroxyl groups, and subsequent coordination of Fe(III) ions to produce an Na2EDTA modified cotton-fiber-Fe complex (Fe-EDTA-Cotton). After characterization by SEM and FTIR, Fe-EDTA-Cotton was used as a heterogeneous photocatalyst to reduce Cr(VI) and degrade Reactive Red 195 oxidatively, to evaluate its catalytic activity. The results indicated that high Na2EDTA and/or NaH2PO4 concentrations, and/or high hydrothermal temperature increased the carboxyl group content (QCOOH) of the cotton fiber; high Fe content and elevated temperature could enhance coordination. Fe-EDTA-Cotton had a significant photocatalytic function in reducing Cr(VI). Increasing the QFe value of the complex or stronger irradiation favored photocatalytic reduction of Cr(VI) species. Increasing the system's pH did not help the reduction reaction. Fe-EDTA-Cotton could also remove Cr(VI) and Reactive Red 195 simultaneously from aqueous solution, and showed strong photocatalytic capacity. The reduction or oxidation reaction could be regulated by changing Na2S2O8 concentration in the reaction system, which might provide a new way to treat wastewater containing organic pigments and Cr(VI) species. Highlight Hydrothermal process enhanced EDTA modification of cotton fabric EDTA-modified cotton fiber Fe complex was prepared Cr(VI) removal was improved by its synergistic effect Fe complexes can remove Cr (VI) and organic dyes from aqueous solution in one bath The work might provide a new way to purify the wastewater containing organic pigments and Cr (VI) species.

Background: Neuromedin B (Nmb) is implicated in the regulation of nociception of sensory neurons. However, the underlying cellular and molecular mechanisms remain unknown. Methods: Using patch clamp recording, western blot analysis, immunofluorescent labelling, enzyme-linked immunosorbent assays, adenovirus-mediated shRNA knockdown and animal behaviour tests, we studied the effects of Nmb on the sensory neuronal excitability and peripheral pain sensitivity mediated by Cav3.2 T-type channels. Results: Nmb reversibly and concentration-dependently increased T-type channel currents (IT) in small-sized trigeminal ganglion (TG) neurons through the activation of neuromedin B receptor (NmbR). This NmbR-mediated IT response was Gq protein-coupled, but independent of protein kinase C activity. Either intracellular application of the QEHA peptide or shRNA-mediated knockdown of Gβ abolished the NmbR-induced IT response. Inhibition of protein kinase A (PKA) or AMP-activated protein kinase (AMPK) completely abolished the Nmb-induced IT response. Analysis of phospho-AMPK (p-AMPK) revealed that Nmb significantly activated AMPK, while AMPK inhibition prevented the Nmb-induced increase in PKA activity. In a heterologous expression system, activation of NmbR significantly enhanced the Cav3.2 channel currents, while the Cav3.1 and Cav3.3 channel currents remained unaffected. Nmb induced TG neuronal hyperexcitability and concomitantly induced mechanical and thermal hypersensitivity, both of which were attenuated by T-type channel blockade. Moreover, blockade of NmbR signalling prevented mechanical hypersensitivity in a mouse model of complete Freund's adjuvant-induced inflammatory pain, and this effect was attenuated by siRNA knockdown of Cav3.2. Conclusions: Our study reveals a novel mechanism by which NmbR stimulates Cav3.2 channels through a Gβγ-dependent AMPK/PKA pathway. In mouse models, this mechanism appears to drive the hyperexcitability of TG neurons and induce pain hypersensitivity.

Objective: The Coronavirus disease 2019 (COVID-19) vaccine is currently available. This timely survey was conducted to provide insight into on the willingness of healthcare workers (HCWs)to receive the vaccine and determine the influencing factors.Methods: This was a cross-sectional online survey. An online questionnaire was provided to all participants and they were asked if they would accept a free vaccine. The questionnaire gathered general demographic information, and included the General Health Questionnaire (GHQ-12); Myers-Briggs Type Indicator questionnaire (MBTI); Depression, Anxiety, and Stress Scales (DASS-21); and the 12-item Short Form Health Survey (SF-12). The data were collected automatically and electronically. Univariate analysis was done between all the variables and our dependent variable. Multivariable logistic regression models were employed to examine and identify the associations between the acceptance of the COVID-19 vaccine with the associated variables.Results: We collected 505 complete answers. The participants included 269 nurses (53.27%), 206 clinicians (40.79%), 15 administrative staff (2.97%), and 15 other staff (2.97%). Of these, 76.63% declared they would accept the vaccine. The major barriers were concerns about safety, effectiveness, and the rapid mutation in the virus. Moreover, four factors were significantly associated with the willingness to receive the vaccine: (a) “understanding of the vaccine” (odds ratio (OR):2.322; 95% confidence interval [CI]: 1.355 to 3.979); (b) “worried about experiencing COVID-19” (OR 1.987; 95% CI: 1.197–3.298); (c) “flu vaccination in 2020” (OR 4.730; 95% CI: 2.285 to 9.794); and (d) “living with elderly individuals” (OR 1.928; 95% CI: 1.074–3.462).Conclusions: During the vaccination period, there was still hesitation in receiving the vaccine. The results will provide a rationale for the design of future vaccination campaigns and education efforts concerning the vaccine.

Aminoglycoside antibiotics, such as gentamicin, are known to have vestibulotoxic effects, including ataxia and disequilibrium. To date, however, the underlying cellular and molecular mechanisms are still unclear. In this study, we determined the role of gentamicin in regulating the sustained delayed rectifier K

Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) is of important clinical significance. In this study, a novel aptamer-based fluorometric assay was developed for detection of MRSA in clinical samples by coupling with immunomagnetic separation . The S. aureus cells in clinical specimens were enriched by magnetic separation. Following lysis by staphylococcal lysin , the PBP2a proteins were released from S. aureus cells and detected by the aptamer-based fluorometric assay. Without lengthy period of bacteria cultivation in the traditional susceptibility testing, this test has an overall testing time of only 2 h with the detection limit of 2.63 × 103 and 1.38 × 103 CFU/mL in PBS and spiked nasal swab, respectively. Since it is simple, rapid and sensitive, this method could be used for the detection of MRSA in various clinical samples.

Nitrogen plays an important role in plant tolerance to salt stress, but it remains largely unknown how the young and old leaves in woody plants differently respond to salinity under different nitrogen supplies. In this study, the young saplings of Populus bolleana Lauche were exposed to moderate salt condition and different nitrogen levels, and the young and old leaves were separately analysed to compare their responses in ion balance, carbon/nitrogen balance, and redox balance. NaCl stress alone profoundly affected the old leaves but only exerted mild effects on young leaves, showing in the former lower K+/Na+ ratio, ion imbalance, lipid peroxidation, chlorophyll degradation, and reduction of carbon and nitrogen concentration. There is an important strategy for the effective distribution of nutrition (K+ and NO3−) to young leaves when P. bolleana suffer salt stress, displayed by the differential regulation of K+ / Na+ and NO3- transporters across leaf ages. When salt stress was accompanied by nitrogen deficiency, the young leaves displayed similar damaged effects with old leaves, with a comparable K+/Na+ ratio in all the damaged leaves. Conversely, high nitrogen supply modestly alleviated salt toxicity in both leaf type, accompanying with modest change of K+/Na+ concentrations. The present study demonstrated that K+/Na+ ratio plays a vital role against salt stress across nitrogen levels in both young and old leaves, and ion imbalance in young leaves can serve as an indicator related to the survival of plants under salt stress. Nitrogen deficiency significantly endanger the survival of salt-stressed poplar for its protection strategy on the young leaves has been lost in this condition.

Objective To compare the outcomes of patients with non-traumatic cardiac arrest (CA) who received early versus late mechanical cardiopulmonary resuscitation (CPR) with the Lund University Cardiac Assist System (LUCAS) device in the emergency department (ED). Methods This was a retrospective observational study in the ED of a single medical center performed from May 2018 to December 2019; 68 patients with CA were eligible. We grouped the patients according to the time to initiating LUCAS use after CA into an early group (≤4 minutes) and late group (>4 minutes). Results The rate of return of spontaneous circulation (ROSC) was higher in the early group vs the late group (69.2% vs 52.4%, respectively). The 4-hour survival rate was significantly higher in the early group vs the late group (83.3% vs 45.5%, respectively), and CPR duration was significantly shorter in the early group (23.3 ± 12.5 vs 31.1 ± 14.8 minutes, respectively). Conclusion Early mechanical CPR can improve the success of achieving ROSC and the 4-hour survival rate in patients with non-traumatic CA in the ED, considering that more benefits were observed in patients who received early vs late LUCAS device therapy.

The present study aimed to investigate the role of homeobox B4 (HOXB4) in breast cancer. Analysis of The Cancer Genome Atlas data revealed that HOXB4 expression was positively associated with expression of the StAR-related lipid transfer domain protein 13 (STARD13), and the overall survival of patients with breast cancer. Immunohistochemistry and quantitative polymerase chain reaction assays demonstrated that HOXB4 expression was downregulated in breast cancer tissues compared with adjacent normal tissues and was additionally positively associated with STARD13 expression. HOXB4 promoted STARD13 expression in breast cancer cells. Chromatin immunoprecipitation and luciferase reporter assays confirmed that HOXB4 directly bound to the STARD13 promoter. Additionally, HOXB4 inhibited breast cancer cell migration and the epithelial-mesenchymal transition through the STARD13/Ras homolog (Rho) family member A/Rho associated protein kinase signaling pathway. HOXB4 overexpression enhanced the sensitivity of breast cancer cells to doxorubicin and reversed resistance in doxorubicin-resistant cells. Overall, the results indicated that HOXB4 inhibited breast cancer cell migration and enhanced the sensitivity of breast cancer cells to doxorubicin by targeting STARD13.

Based on imidazo[1,2- a ]pyridine derivatives, a series of mononuclear and binuclear fluorine-boron complexes 1a , b , 4a , b , and 5a , b have been designed and synthesized. X-ray crystal structures for single boron-core complexes 1b , 4a , 4b , and dual boron-core imidazo[1,2- a ]pyridine-BODIPY dyad 5b were obtained, and intermolecular C H⋯F interactions were observed in their solid state. High absorption coefficients, strong emissions, and high photostabilities have been demonstrated for these boron complexes. DFT and TD-DFT studies reveal efficient electronic interactions between the imidazo[1,2- a ]pyridine moiety and BODIPY plane for dyads 4a , b , and 5a , b . Cell imaging of 4a was tested to show good cellular uptake ability.

Phosphatidylcholine (PC) is a rare membrane lipid in bacteria but crucial for virulence of various plant and animal pathogens. The pcs- mutant lacking PC in bacterial membranes of Pseudomonas syringae pv. syringae van Hall 1336 displayed more ampicillin resistance. Ampicillin susceptibility tests gave an IC50 (half maximal inhibitory concentration) of 52 mg/ml for Pseudomonas syringae pv. syringae van Hall 1336, 53 mg/ml for the complemented strain 1336 RM (pcs-/+) and 90 mg/ml for the 1336 pcs- mutant. Activity assay of β-lactamase in periplasmic extracts gave 0.050 U/mg for the 1336 wild type, 0.052 U/mg for the 1336RM (pcs-/+), 0.086 U/mg for the 1336 pcs- mutant. Analysis by western blotting showed that the content of AmpC enzyme was markedly different in periplasmic extracts between the wild-type and pcs- mutant strains. Reverse transcriptase PCR also showed that the presence or absence of PC in bacterial membranes did not affect the transcription of ampC gene. The phenotype of the pcs- mutant was able to be recovered to the wild type by introducing a wild-type pcs gene into the pcs- mutant. Similar results were also obtained from the soil-dwelling bacterium Pseudomonas sp. 593. Our results demonstrate that the absence of PC in bacterial membranes facilitates the translocation of Sec-dependent β-lactamase AmpC from cytoplasm to periplasm, and the enhanced ampicillin-resistance in the pcs- strains mainly comes from effective translocation of AmpC via Sec-pathway.

A new freshwater crab of the genus Sinopotamon Bott, 1967, S. baokangense sp. nov. (Potamidae), is described from Baokang County, Hubei Province, P.R. China. The new species closely resembles two other Chinese species: S. cochlearidigitum Dai, Chen, Zhang & Lin, 1986, from Fang and Zhushan Counties; and S. introdigitum Dai, Chen, Zhang & Lin, 1986, from Fang and Yun Counties and Shennongjia, in general morphology and their distinctive spoon-tipped cheliped fingers. The new species, however, can easily be distinguished by a number of cheliped, male abdominal, and male first gonopod characters.

IntroductionLarge artery intracranial occlusive disease including middle cerebral artery (MCA) is a major contributor to the incidence of stroke in China. The data on the prognosis of symptomatic atherosclerotic MCA occlusions (MCAO) are limited. We aimed to investigate the related factors of unfavorable outcomes in patients with stroke associated with MCAO.Material and methodsA total of 119 patients with MCAO symptom were enrolled in this retrospective longitudinal cohort study. All patients met inclusion criteria of cerebral angiography by CT angiography or magnetic resonance angiography. Stroke severity was assessed on admission using the National Institutes of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS).ResultsWe showed an average follow-up time of 46.8 months, within which 20 cases (19.6%) died and 14 cases (13.7%) had stroke recurrence. Using mRS as an evaluation index, the patients were divided into an unfavourable outcome group (mRS > 2, 48 cases) and a favourable outcome group (mRS ≤ 2, 54 cases). Logistic regression analysis suggests that age and NIHSS score were independent risk factors for a poor outcome value. Coexisting other cerebral vascular occlusion was an independent risk factor for stroke recurrence. Age was an independent risk factor for death.ConclusionsThe prognosis of patients with MCAO was generally optimistic, with higher survival rate and longer survival time. As compared, elder age and higher NIHSS score both tend to be associated with worse prognosis of survival. MCAO patients with other extracranial or intracranial vascular occlusion have higher risk of recurrent stroke. Death rate increases with age among the MCAO patients.

Background Migraine is a debilitating neurological disorder involving abnormal trigeminovascular activation and sensitization. However, the underlying cellular and molecular mechanisms remain unclear. Methods A rat model of conscious migraine was established through the electrical stimulation (ES) of the dural mater surrounding the superior sagittal sinus. Using patch clamp recording, immunofluorescent labelling, enzyme-linked immunosorbent assays and western blot analysis, we studied the effects of ES on sensory neuronal excitability and elucidated the underlying mechanisms mediated by voltage-gated ion channels. Results The calcitonin gene-related peptide (CGRP) level in the jugular vein blood and the number of CGRP-positive neurons in the trigeminal ganglia (TGs) were significantly increased in rats with ES-induced migraine. The application of ES increased actional potential firing in both small-sized IB4-negative (IB4 −) and IB4 + TG neurons. No significant changes in voltage-gated Na+ currents were observed in the ES-treated groups. ES robustly suppressed the transient outward K+ current (I A) in both types of TG neurons, while the delayed rectifier K+ current remained unchanged. Immunoblot analysis revealed that the protein expression of Kv4.3 was significantly decreased in the ES-treated groups, while Kv1.4 remained unaffected. Interestingly, ES increased the P/Q-type and T-type Ca2+ currents in small-sized IB4 − TG neurons, while there were no significant changes in the IB4 + subpopulation of neurons. Conclusion These results suggest that ES decreases the I A in small-sized TG neurons and increases P/Q- and T-type Ca2+ currents in the IB4 − subpopulation of TG neurons, which might contribute to neuronal hyperexcitability in a rat model of ES-induced migraine.

Background Cholecystokinin (CCK) is implicated in the regulation of nociceptive sensitivity of primary afferent neurons. Nevertheless, the underlying cellular and molecular mechanisms remain unknown. Methods Using patch clamp recording, western blot analysis, immunofluorescent labelling, enzyme-linked immunosorbent assays, adenovirus-mediated shRNA knockdown and animal behaviour tests, we studied the effects of CCK-8 on the sensory neuronal excitability and peripheral pain sensitivity mediated by A-type K+ channels. Results CCK-8 reversibly and concentration-dependently decreased A-type K+ channel (IA) in small-sized dorsal root ganglion (DRG) neurons through the activation of CCK type B receptor (CCK-BR), while the sustained delayed rectifier K+ current was unaffected. The intracellular subunit of CCK-BR coimmunoprecipitated with Gαo. Blocking G-protein signaling with pertussis toxin or by the intracellular application of anti-Gβ antibody reversed the inhibitory effects of CCK-8. Antagonism of phosphatidylinositol 3-kinase (PI3K) but not of its common downstream target Akts abolished the CCK-BR-mediated IA response. CCK-8 application significantly activated JNK mitogen-activated protein kinase. Antagonism of either JNK or c-Src prevented the CCK-BR-mediated IA decrease, whereas c-Src inhibition attenuated the CCK-8-induced p-JNK activation. Application of CCK-8 enhanced the action potential firing rate of DRG neurons and elicited mechanical and thermal pain hypersensitivity in mice. These effects were mediated by CCK-BR and were occluded by IA blockade. Conclusion Our findings indicate that CCK-8 attenuated IA through CCK-BR that is coupled to the Gβγ-dependent PI3K and c-Src-mediated JNK pathways, thereby enhancing the sensory neuronal excitability in DRG neurons and peripheral pain sensitivity in mice.

From the view point of the automotive multi-value-chain in collaborative businesses, different customers will produce unequal values in the future services. To strive for the continuous contribution of high-value customers, and optimize the collaborative business efficiency in parts supplier value chain and service multi-value-chain, in this paper, we will mention a customer analysis method for service multi-value-chain in automotive industry. Firstly, according to the related data in our automotive service collaborative business platform, we selected some evaluation indicators to describe the customer’s current value and potential value. Then, a customer value analysis model was offered based on a multi-classification SVM, which would give a classification recommendation according to the potential value of customs. Finally, we randomly selected 40 customers’ 36 months related information from our platform, to verify our method by some experiments. The experimental result shows that the customer classification was close to the actual situation. Among the 40 testing samples, the accuracy rate of customer value classification reaches to 87.5%. The results of customer value forecasts can be used to guide parts supplier and service providers to make a more effective policy, such as giving specific offers for outstanding customers. It will expand the synergy of multi-value-chain outside the Three-guarantee period of serving period of automobile service, and realize the strategic management of service life cycle centered on customer value.

Auto parts in the after-sales service flow across parts supplier, manufacturer, and transit store as well as service provider. The accurate and timely demand for auto parts plays a key role in the value-added process of the parts multiple-value chains. This paper presents a collaboration mechanism and evaluation method for auto parts multiple-value chains based on sales prediction. First, we study the composition of inner value chain of independent accounting unit, and discuss factors of value increment of the inner value chain. Moreover, dominated elements are selected as main factors for future value. Second, a colored petri nets model is created for each independent accounting unit all over the multiple-value-chains, of which value-added process maps to Transition, repository to Place and auto parts to Place’s Token. Such model is utilized to simulate the value-added process over the multiple-value chains. Finally, a collaboration mechanism for multiple-value chains is designed based on the prediction amount for auto parts. By means of randomly generating a sequence of custom’s arrival and Monte Carlo method, we use CPN Tools to simulate the value-added process over the multiple-value chains. Experiments show that the offered auto parts collaboration mechanism can fully utilize existed resources in the multiple-value chains, gets maximized rise in value, and further seek the fundamental path to value increment. The model implemented in this paper is able to provide some quantification references for the multiple-value chains.

Aminoglycoside antibiotics, such as gentamicin, are known to have vestibulotoxic effects, including ataxia and disequilibrium. To date, however, the underlying cellular and molecular mechanisms are still unclear. In this study, we determined the role of gentamicin in regulating the sustained delayed rectifier K+ current (IDR) and membrane excitability in vestibular ganglion (VG) neurons in mice. Our results showed that the application of gentamicin to VG neurons decreased the IDR in a concentration-dependent manner, while the transient outward A-type K+ current (IA) remained unaffected. The decrease in IDR induced by gentamicin was independent of G-protein activity and led to a hyperpolarizing shift of the inactivation Vhalf. The analysis of phospho-c-Jun N-terminal kinase (p-JNK) revealed that gentamicin significantly stimulated JNK, while p-ERK and p-p38 remained unaffected. Blocking Kv1 channels with α-dendrotoxin or pretreating VG neurons with the JNK inhibitor II abrogated the gentamicin-induced decrease in IDR. Antagonism of JNK signaling attenuated the gentamicin-induced stimulation of PKA activity, whereas PKA inhibition prevented the IDR response induced by gentamicin. Moreover, gentamicin significantly increased the number of action potentials fired in both phasic and tonic firing type neurons; pretreating VG neurons with the JNK inhibitor II and the blockade of the IDR abolished this effect. Taken together, our results demonstrate that gentamicin decreases the IDR through a G-protein-independent but JNK and PKA-mediated signaling pathways. This gentamicin-induced IDR response mediates VG neuronal hyperexcitability and might contribute to its pharmacological vestibular effects.

Sinopotamon Bott, 1967 is the most speciose and widely distributed freshwater crab genus in East Asia. Our extensive sampling includes about 76% of the known Sinopotamon taxa, and nearly covers its entire distribution area. Based on mitochondrial cytochrome oxidase I (COI) and 16S rRNA, as well as nuclear 28S rRNA and histone H3, we reconstructed the Sinopotamon phylogeny using maximum likelihood and Bayesian approaches. The divergence time was estimated and multiple methods were used to conduct diversification analyses. The ancestral geographic distribution and character state were reconstructed. Three main clades (Clades I, II and III) that roughly correspond to their main geographic distribution ranges were recovered. Our results challenge the current view of the four major species groups based on the morphological differences in the male first gonopod (G1). The most recent common ancestor of Sinopotamon most likely originated from the Sichuan Basin and surrounding mountains (SBSM) and subsequently dispersed throughout central and eastern China. The exceptionally rapid, recent diversification was detected in Clade II. The high incidence of species-level non-monophyly found in Clade II can be explained by recent rapid radiation. Climatic changes, morphological innovations, range expansion and geographical heterogeneity may all contribute to the diversification in Sinopotamon. This study contributes to our knowledge on diversification of freshwater benthic macro-invertebrates in the East Asian inland ecosystem.

The knowledge regarding the effects of metal or metal oxide nanoparticles on soil microbial metabolic activity and key ecological functions is limited, relative to the information about their species diversity. For this reason, the responses of soil microbial metabolic activity to silver (AgNPs) and iron oxide (FeONPs) nanoparticles, along concentration gradients of each, were evaluated by microcalorimetry and soil nitrification potential. The changes in abundances of bacteria, eukaryotes and ammonia-oxidizing bacteria were measured by real time quantitative PCR. It was found that AgNP (at 0.1, 1 and 10 mg kg(-1) soil) amendments decreased soil microbial metabolic activity, nitrification potential and the abundances of bacteria and ammonia-oxidizing bacteria; on the contrary, FeONPs had the positive effects on soil microbial metabolic activity (at 1 and 10 mg kg(-1) soil) and soil nitrification potential (at 0.1 and 1 mg kg(-1) soil). Specific microbial metabolic activity and specific nitrification potential further revealed that metal or metal oxide nanoparticles could change the C and N cycles of the agricultural soil through influencing soil microbial metabolism. These findings could deepen the understanding of the influence of NPs on soil microorganisms and their driven soil ecology process.

Introduction: Internal carotid artery occlusion (ICAO) causes high annual rates of mortality and morbidity. It has been established that atherosclerosis is the normal cause of ICAO. As the pathogenesis of atherosclerosis may involve blood lipids, inflammatory factors and other biomarkers, the aim of this study was to assess the changes in these biomarkers and investigate the relationship between these biomarkers and the development of ICAO in stroke patients. Material and methods: A total of 89 ischaemic stroke inpatients with ICAO (ICAO group) and 89 without ICAO (control group) were studied, retrospectively. The serum was collected from each patient on the 3 rd day of admission, to measure the lipid parameters and biomarkers, e.g. high-sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR), and lactic acid (LA). Histories were taken including age, gender, smoking history, and disease history. Additional analysis was carried out to compare between the genders and evaluate the association between certain biomarkers and ICAO. Results: Among the 89 ICAO cases in this study, the serum levels of hs-CRP, ESR and LA were significantly higher than those in the control group (p ≤ 0.001). No significant differences were found in the mean levels of total cholesterol, triacylglycerol, HDL cholesterol or glucose, or the known risk factors. Gender also had no influence on these biomarkers. Logistic regression analysis indicated that hs-CRP, ESR and LA were significantly associated with ICAO (p ≤ 0.05). Conclusions: These results suggest that hs-CRP, ESR and LA are associated with ICAO in ischaemic stroke patients, but gender has no effect. Therefore, Hs-CRP, ESR and LA may be useful in the early detection of patients with ICAO.

α-Melanocyte–stimulating hormone (α-MSH) has been shown to be involved in nociception, but the underlying molecular mechanisms remain largely unknown. In this study, we report that α-MSH suppresses the transient outward A-type K(+) current (I(A)) in trigeminal ganglion (TG) neurons and thereby modulates neuronal excitability and peripheral pain sensitivity in rats. Exposing small-diameter TG neurons to α-MSH concentration-dependently decreased I(A). This α-MSH–induced I(A) decrease was dependent on the melanocortin type 4 receptor (MC4R) and associated with a hyperpolarizing shift in the voltage dependence of A-type K(+) channel inactivation. Chemical inhibition of phosphatidylinositol 3-kinase (PI3K) with wortmannin or of class I PI3Ks with the selective inhibitor CH5132799 prevented the MC4R-mediated I(A) response. Blocking G(i/o)-protein signaling with pertussis toxin or by dialysis of TG neurons with the G(βγ)-blocking synthetic peptide QEHA abolished the α-MSH–mediated decrease in I(A). Further, α-MSH increased the expression levels of phospho-p38 mitogen-activated protein kinase, and pharmacological or genetic inhibition of p38α abrogated the α-MSH–induced I(A) response. Additionally, α-MSH significantly increased the action potential firing rate of TG neurons and increased the sensitivity of rats to mechanical stimuli applied to the buccal pad area, and both effects were abrogated by I(A) blockade. Taken together, our findings suggest that α-MSH suppresses I(A) by activating MC4R, which is coupled sequentially to the G(βγ) complex of the G(i/o)-protein and downstream class I PI3K-dependent p38α signaling, thereby increasing TG neuronal excitability and mechanical pain sensitivity in rats.

Glycophytic plants suffer from severe stress and injury when roots are exposed to high salinity in the rhizosphere. In contrast, the euhalophyte Salicornia europaea grows well at 200 mM NaCl and can withstand up to 1000 mM NaCl in the root zone. Analysis of gene expression profiles and the underlying molecular mechanisms responsible for this tolerance have been largely overlooked. Using the Illumina sequencing platform and the short-reads assembly programme Trinity, we generated a total of 40 and 39 million clean reads and further 140,086 and 122,728 unigenes from the 200 mM NaCl and 0 mM NaCl treated tissues of S. europaea roots, respectively. All unigenes in this study were functionally annotated within context of the COG, GO and KEGG pathways. Unigene functional annotation analysis allowed us to identify hundreds of ion transporters related to homeostasis and osmotic adaptation as well as a variety of proteins related to cation, amino acid, lipid and sugar transport. We found significant enrichment in response to stress including the functional categories of “antioxidant activity”, “catalytic activity” and “response to stimuli”. These findings represent for a useful resource for the scientific community working on salt tolerance mechanisms. Conversely, a total of 8639 EST-SSRs from 131,594 unigenes were identified and 4539 non-redundant SSRs primers pairs were developed. These data provide a good foundation for future studies on molecular adaptation mechanisms of euhalophytes roots under saline environments and will likely facilitate the identification of critical salt tolerance traits to be transferred in economically important crops.

Additional file 1: Figure S1. Recombination hotspots in 11 LGs. Figure S2. Distribution of r2 values between markers for Jatropha. Figure S3. Genome-wide LD decay (mean r2) estimated using 3422 markers that are mapped on 11 LGs of Jatropha ultra-high density genetic map. Table S1. Recombination hotspots in 11 LGs. Table S2. Genome-wide LD estimates among marker-pairs mapped across 11 LGs of ultra-high density genetic map for Jatropha.

Recent studies implicate melatonin in the antinociceptive activity of sensory neurons. However, the underlying mechanisms are still largely unknown. Here, we identify a critical role of melatonin in functionally regulating Cav3.2 T-type Ca

Objective This study aimed to reveal the mitochondrial genomes (mtgenomes) of Tetrix japonica and Alulatettix yunnanensis, and the phylogenetics of Orthoptera species. Methods The mtgenomes of A. yunnanensis and T. japonica were firstly sequenced and assembled through partial sequences amplification, and then the genome organization and gene arrangement were analyzed. Based on nucleotide/amino acid sequences of 13 protein-coding genes and whole mtgenomes, phylogenetic trees were established on 37 Orthoptera species and 5 outgroups, respectively. Results Except for a regulation region (A+T rich region), a total of 37 genes were found in mtgenomes of T. japonica and A. yunnanensis, including 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes, which exhibited similar characters with other Orthoptera species. Phylogenetic tree based on 13 concatenated protein-coding nucleotide sequences were considered to be more suitable for phylogenetic reconstruction of Orthoptera species than amino acid sequences and mtgenomes. The phylogenetic relationships of Caelifera species were Acridoidea and Pamphagoidea > Pyrgomorphoidea > Pneumoroidea > Eumastacoidea > Tetrigoidea > Tridactyloidea. Besides, a sister-group relationship between Tettigonioidea and Rhaphidophoroidea was revealed in Ensifera. Conclusion Concatenated protein-coding nucleotide sequences of 13 genes were suitable for reconstruction of phylogenetic relationship in orthopteroid species. Tridactyloidea was a sister group of Tetrigoidea in Caelifera, and Rhaphidophoroidea was a sister group of Tettigonioidea in Ensifera. Electronic supplementary material The online version of this article (doi:10.1186/s40659-017-0132-9) contains supplementary material, which is available to authorized users.

Background As an important biofuel plant, the demand for higher yield Jatropha curcas L. is rapidly increasing. However, genetic analysis of Jatropha and molecular breeding for higher yield have been hampered by the limited number of molecular markers available. Results An ultrahigh-density linkage map for a Jatropha mapping population of 153 individuals was constructed and covered 1380.58 cM of the Jatropha genome, with average marker density of 0.403 cM. The genetic linkage map consisted of 3422 SNP and indel markers, which clustered into 11 linkage groups. With this map, 13 repeatable QTLs (reQTLs) for fruit yield traits were identified. Ten reQTLs, qNF-1, qNF-2a, qNF-2b, qNF-2c, qNF-3, qNF-4, qNF-6, qNF-7a, qNF-7b and qNF-8, that control the number of fruits (NF) mapped to LGs 1, 2, 3, 4, 6, 7 and 8, whereas three reQTLs, qTWF-1, qTWF-2 and qTWF-3, that control the total weight of fruits (TWF) mapped to LGs 1, 2 and 3, respectively. It is interesting that there are two candidate critical genes, which may regulate Jatropha fruit yield. We also identified three pleiotropic reQTL pairs associated with both the NF and TWF traits. Conclusion This study is the first to report an ultrahigh-density Jatropha genetic linkage map construction, and the markers used in this study showed great potential for QTL mapping. Thirteen fruit-yield reQTLs and two important candidate genes were identified based on this linkage map. This genetic linkage map will be a useful tool for the localization of other economically important QTLs and candidate genes for Jatropha. Electronic supplementary material The online version of this article (10.1186/s13068-017-1004-9) contains supplementary material, which is available to authorized users.

As an indispensable part of Internet of Things (IoT), wireless sensor networks (WSNs) are more and more widely used with the rapid development of IoT. The neighbor discovery protocols are the premise of communication between nodes and networking in energy-limited self-organizing wireless networks, and play an important role in WSNs. Because the node energy is limited, neighbor discovery must operate in an energy-efficient manner, that is, under the condition of a given energy budget, the neighbor discovery performance should be as good as possible, such that the discovery latency would be as small as possible and the discovered neighbor percentage as large as possible. The indirect neighbor discovery mainly uses the information of the neighbors that have been found by a pairwise discovery method to more efficiently make a re-planning of the discovery wake-up schedules of the original pairwise neighbor discovery, thereby improving the discovery energy efficiency. The current indirect neighbor discovery methods are mainly divided into two categories: one involves removing the inefficient active slots in the original discovery wake-up schedules, and the other involves adding some efficient active slots. However, the two categories of methods have their own limitations. The former does not consider that this removal operation destroys the integrity of the original discovery wake-up schedules and hence the possibility of discovering new neighbors is reduced, which adversely affects the discovered neighbor percentage. For the latter category, there are still inefficient active slots that were not removed in the re-planned wake-up schedules. The motivation of this paper is to combine the advantages of these two types of indirect neighbor discovery methods, that is, to combine the addition of efficient active slots and the removal of inefficient active slots. To achieve this goal, this paper proposes, for the first time, the concept of virtual nodes in neighbor discovery to maximize the integrity of the original wake-up schedules and achieve the goals of adding efficient active slots and removing inefficient active slots. Specifically, a virtual node is a collaborative group that is formed by nodes within a small range. The nodes in a collaborative group share responsibility for the activating task of one member node, and the combination of these nodes&rsquo, wake-up schedules forms the full wake-up schedule of a node that only uses a pairwise method. In addition, this paper proposes a set of efficient group management mechanisms, and the key steps affecting energy efficiency are analyzed theoretically to obtain the energy-optimal parameters. The extended simulation experiments in multiple scenarios show that, compared with other methods, our neighbor discovery protocol based on virtual nodes (VN-NDP) has a significant improvement in average discovery delay and discovered neighbor percentage performance at a given energy budget. Compared with the typical indirect neighbor discovery algorithm EQS, a neighbor discovery with extended quorum system, our proposed VN-NDP method reduces the average discovery delay by up to 10 . 03 % and increases the discovered neighbor percentage by up to 18 . 35 % .

Authorship attribution refers to the task of automatically determining the author based on a given sample of text. It is a problem with a long history and has a wide range of application. Building author profiles using language models is one of the most successful methods to automate this task. New language modeling methods based on neural networks alleviate the curse of dimensionality and usually outperform conventional N-gram methods. However, there have not been much research applying them to authorship attribution. In this paper, we present a novel setup of a Neural Network Language Model (NNLM) and apply it to a database of text samples from different authors. We investigate how the NNLM performs on a task with moderate author set size and relatively limited training and test data, and how the topics of the text samples affect the accuracy. NNLM achieves nearly 2.5% reduction in perplexity, a measurement of fitness of a trained language model to the test data. Given 5 random test sentences, it also increases the author classification accuracy by 3.43% on average, compared with the N-gram methods using SRILM tools. An open source implementation of our methodology is freely available at https://github.com/zge/authorship-attribution/., International Conference on Pattern Recognition Application and Methods (ICPRAM) 2016

Recent studies implicate melatonin in the antinociceptive activity of sensory neurons. However, the underlying mechanisms are still largely unknown. Here, we identify a critical role of melatonin in functionally regulating Cav3.2 T-type Ca2+ channels (T-type channel) in trigeminal ganglion (TG) neurons. Melatonin inhibited T-type channels in small TG neurons via the melatonin receptor 2 (MT2 receptor) and a pertussis toxin-sensitive G-protein pathway. Immunoprecipitation analyses revealed that the intracellular subunit of the MT2 receptor coprecipitated with Gαo . Both shRNA-mediated knockdown of Gαo and intracellular application of QEHA peptide abolished the inhibitory effects of melatonin. Protein kinase C (PKC) antagonists abolished the melatonin-induced T-type channel response, whereas inhibition of conventional PKC isoforms elicited no effect. Furthermore, application of melatonin increased membrane abundance of PKC-eta (PKCη ) while antagonism of PKCη or shRNA targeting PKCη prevented the melatonin-mediated effects. In a heterologous expression system, activation of MT2 receptor strongly inhibited Cav3.2 T-type channel currents but had no effect on Cav3.1 and Cav3.3 current amplitudes. The selective Cav3.2 response was PKCη dependent and was accompanied by a negative shift in the steady-state inactivation curve. Furthermore, melatonin decreased the action potential firing rate of small TG neurons and attenuated the mechanical hypersensitivity in a mouse model of complete Freund's adjuvant-induced inflammatory pain. These actions were inhibited by T-type channel blockade. Together, our results demonstrated that melatonin inhibits Cav3.2 T-type channel activity through the MT2 receptor coupled to novel Gβγ -mediated PKCη signaling, subsequently decreasing the membrane excitability of TG neurons and pain hypersensitivity in mice.

A total of twenty microsatellite loci included dinucleotide, trinucleotide, and tetranucleotide microsatellite loci were isolated for Sinopotamon yangtsekiense using the FIASCO protocol. Genotyping of 30 individuals showed that the number of alleles ranged from 2 to 22, with observed heterozygosity ranging from 0.103 to 0.900 and expected heterozygosity ranging from 0.100 to 0.957. The polymorphism information content per locus ranged from 0.093 to 0.932. Cross-species amplification was investigated in seven related taxa. The results suggested that the developed microsatellite loci are specific to Sinopotamon and have potential for limited use in cross-genus amplification within Potamidae. These microsatellite markers should prove valuable tools for population genetic studies of the species Sinopotamon.