SNAP25/syntaxin4/VAMP2/Munc18-1 Complexes in Spinal Dorsal Horn Contributed to Inflammatory Pain.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) have been implicated in the trafficking of postsynaptic glutamate receptors, including N-methyl-d-aspartate (NMDA)-subtype glutamate receptors (NMDARs) that are critical for nociceptive plasticity and behavioral sensitization. However, the components of SNAREs complex involved in spinal nociceptive processing remain largely unknown. Here we found that SNAP25, syntaxin4, VAMP2 and Munc18-1 were localized at postsynaptic sites and formed the complex in the superficial lamina of spinal cord dorsal horn of rats. The complex formation between these SNAREs components were accelerated after intraplantar injection of complete Freund's adjuvant (CFA), pharmacological removal of GABAergic inhibition or activation of NMDAR in intact rats. The increased SNAP25/syntaxin4/VAMP2/Munc18-1 interaction facilitated the surface delivery and synaptic accumulation of NMDAR during inflammatory pain. Disruption of the molecular interaction between SNAP25 with its SNARE partners by using a blocking peptide derived from the C-terminus of SNAP25 effectively repressed the surface and synaptic accumulation of GluN2B-containing NMDARs in CFA-injected rats. This peptide also alleviated inflammatory mechanical allodynia and thermal hypersensitivity. These data suggested that SNAREs complex assembly in spinal cord dorsal horn was involved in the inflammatory pain hypersensitivity through promoting NMDAR synaptic trafficking.

mGluR5/ERK signaling regulated the phosphorylation and function of glycine receptor α1ins subunit in spinal dorsal horn of mice.

Inhibitory glycinergic transmission in adult spinal cord is primarily mediated by glycine receptors (GlyRs) containing the α1 subunit. Here, we found that α1ins, a longer α1 variant with 8 amino acids inserted into the intracellular large loop (IL) between transmembrane (TM)3 and TM4 domains, was expressed in the dorsal horn of the spinal cord, distributed at inhibitory synapses, and engaged in negative control over nociceptive signal transduction. Activation of metabotropic glutamate receptor 5 (mGluR5) specifically suppressed α1ins-mediated glycinergic transmission and evoked pain sensitization. Extracellular signal-regulated kinase (ERK) was critical for mGluR5 to inhibit α1ins. By binding to a D-docking site created by the 8-amino-acid insert within the TM3-TM4 loop of α1ins, the active ERK catalyzed α1ins phosphorylation at Ser380, which favored α1ins ubiquitination at Lys379 and led to α1ins endocytosis. Disruption of ERK interaction with α1ins blocked Ser380 phosphorylation, potentiated glycinergic synaptic currents, and alleviated inflammatory and neuropathic pain. These data thus unraveled a novel, to our knowledge, mechanism for the activity-dependent regulation of glycinergic neurotransmission.

A synthetic peptide disturbing GluN2A/SHP1 interaction in dorsal root ganglion attenuated pathological pain.

Src Homology 2 domain-containing protein tyrosine phosphatase 1 (SHP1) interacts specifically with GluN2A subunit of N-methyl-D-aspartate (NMDA) subtype of glutamate receptors in spinal cord dorsal horn. This molecular interaction is involved in the development of GluN2A-dependent spinal sensitization of nociceptive behaviors. Intrathecal application of a GluN2A-derived polypeptide (short for pep-GluN2A) has been shown to disturb spinal GluN2A/SHP1 interaction and inhibit inflammatory pain. Here we found that SHP1 was also located at dorsal root ganglion (DRG) neurons and formed complexes with GluN2A subunit. Peripheral inflammation activated SHP1 in DRG neurons, which promoted GluN2A tyrosine phosphorylation. The SHP1 binding to GluN2A facilitated the glutamate release from primary afferent fibers and exaggerated nociceptive synaptic transmission onto postsynaptic spinal cord neurons. Our data showed that intradermal application of pep-GluN2A disrupted GluN2A/SHP1 interaction in DRG neurons, attenuated the ability of GluN2A subunit-containing NMDA receptors to regulate the presynaptic glutamate release and more importantly, alleviated the pain hypersensitivity caused by carrageenan, complete Freund's adjuvant and formalin. The neuropathic pain induced by spared nerve injury was also ameliorated by intradermal pep-GluN2A application. These data suggested that disruption of GluN2A/SHP1 interaction in DRG neurons generated an effective analgesic action against pathological pain.

Ubiquitination and inhibition of glycine receptor by HUWE1 in spinal cord dorsal horn.

Glycine receptors (GlyRs) are pentameric proteins that consist of α (α1-α4) subunits and/or ß subunit. In the spinal cord of adult animals, the majority of inhibitory glycinergic neurotransmission is mediated by α1 subunit-containing GlyRs. The reduced glycinergic inhibition (disinhibition) is proposed to increase the excitabilities and spontaneous activities of spinal nociceptive neurons during pathological pain. However, the molecular mechanisms by which peripheral lesions impair GlyRs-α1-mediated synaptic inhibition remain largely unknown. Here we found that activity-dependent ubiquitination of GlyRs-α1 subunit might contribute to glycinergic disinhibition after peripheral inflammation. Our data showed that HUWE1 (HECT, UBA, WWE domain containing 1), an E3 ubiquitin ligase, located at spinal synapses and specifically interacted with GlyRs-α1 subunit. By ubiquitinating GlyRs-α1, HUWE1 reduced the surface expression of GlyRs-α1 through endocytic pathway. In the dorsal horn of Complete Freund's Adjuvant-injected mice, shRNA-mediated knockdown of HUWE1 blunted GlyRs-α1 ubiquitination, potentiated glycinergic synaptic transmission and attenuated inflammatory pain. These data implicated that ubiquitin modification of GlyRs-α1 represented an important way for peripheral inflammation to reduce spinal glycinergic inhibition and that interference with HUWE1 activity generated analgesic action by resuming GlyRs-α1-mediated synaptic transmission.

Activity-dependent Synaptic Recruitment of Neuroligin 1 in Spinal Dorsal Horn Contributed to Inflammatory Pain.

Neuroligin 1 (NLGN1), a cell adhesion molecule present at excitatory glutamatergic synapses, has been shown to be critical for synaptic specialization and N-methyl-d-aspartate (NMDA)-subtype glutamate receptor-dependent synaptic plasticity. Whether and how NLGN1 is engaged in nociceptive behavioral sensitization remains largely unknown. In this study, we found an activity-dependent regulation of NLGN1 synaptic expression in pain-related spinal cord dorsal horns of mice. The enhancement of neuronal activity by pharmacological activation of NMDA receptor (NMDAR) or removal of GABAergic inhibition in intact mice significantly increased NLGN1 concentration at synaptosomal membrane fraction. Intraplantar injection of complete Freund's adjuvant (CFA) also increased the NLGN1 expression at synapses. NMDAR might act through Ca2+/calmodulin-dependent protein kinase II (CaMKII) and Src-family protein tyrosine kinase member Fyn to induce the synaptic redistribution of NLGN1. We also found that one of the important roles of NLGN1 was to facilitate the clustering of NMDAR at synapses. The NLGN1-targeting siRNA suppressed the synaptic expression of GluN2B-containing NMDAR in CFA-injected mice and meanwhile, attenuated the inflammatory mechanical allodynia and thermal hypersensitivity. These data suggested that tissue injury-induced synaptic redistribution of NLGN1 was involved in the development of pain hypersensitivity through facilitating the synaptic incorporation of NMDARs.

[Effects of precipitation changes on the precipitation use efficiency and aboveground productivity of alpine steppe-meadow on northern Tibetan Plateau, China.] / 降水变化对藏北高寒草原化草甸降水利用效率及地上生产力的影响.

Total amount, distribution pattern and occurrence time of precipitation determine the water condition of alpine vegetation growth on the Tibetan Plateau, China. Precipitation use efficiency (the ratio of aboveground productivity to precipitation, PUE) is an effective indicator for the relationship between precipitation and vegetation productivity. In this study, we calculated aboveground net primary productivity (ANPP) of an alpine steppe-meadow in northern Tibetan Plateau from 2000 to 2016 based on the correlation model between long-term monitoring biomass data and enhanced vegetation index (EVI) in Damxung. Meteorological data during the same period was also analyzed. Growing season precipitation (GSP) represented the amount of precipitation, improved precipitation concentrated index (PCI) indicated their distribution pattern in the growing season, and precipitation centroid (PC) represented the time of precipitation occurrence. Structural equation model was used to explore the effects of climatic factors on PUE and ANPP, with the consideration of growing season temperature (GST). Results showed that ANPP of this alpine steppe-meadow was mainly controlled by precipitation during the growing season. GSP and ANPP showed significantly positive correlation, whereas the correlation of GST with PUE and ANPP was not significant. PCI and PUE showed significantly positive correlation, which indicated that concentrated distribution of precipitation was beneficial to the increase of PUE. Indirect coefficient between PCI and ANPP was greater than direct coefficient, indicating that PCI influenced ANPP through PUE. Changes of precipitation concentrated time did not show significant effect on PUE and ANPP. Our results suggested that under the rapid climate warming on the Tibetan Plateau, future changes of precipitation and their concentration would have important impacts on aboveground productivity of the alpine grassland.

[Effects of warming and nitrogen addition on community production and biomass allocation in an alpine meadow.] / å¢žæ¸©æ–½æ°®å¯¹é«˜å¯’è‰ç”¸ç”Ÿäº§åŠ›åŠç”Ÿç‰©é‡åˆ†é çš„å½±å“.

A long-term experiment, involving exogenous N addition and simulated warming, was conducted in an alpine meadow in Damxung, northern Tibet, to study how warming and N addition influence community production and biomass allocation. The results showed that warming resulted in a warm but dry microsite, that was, air temperature increased by 1.6 ℃ and soil surface temperature increased by 1.4 ℃, and soil water content decreased by 4.7%. Under no N addition treatments, warming significantly decreased plant aboveground biomass by 61.5%, 108.8% and 77.1% in 2012, 2013 and 2014, respectively. Under high N treatments (40 and 80 kg N·hm-2·a-1), warming had no significant effect on aboveground biomass. These findings indicated that the effect of warming might be dependent on N addition level, and N addition could compensate for soil N loss caused by warming. Warming led to an increase in root/shoot by 98.6%, 60.7% and 97.8% in 2012, 2013 and 2014 under no N addition treatments, respectively. Under the ambient condition, the change percentages of aboveground and belowground biomass of plant communities first increased and then decreased along an N gradient, with the saturation thresholds of above- and below-ground biomass for N addition 56.0 and 55.5 kg N·hm-2·a-1, respectively. Under the warming condition, above- and belowground biomass increased linearly with increasing N addition. These findings suggested that warming modulated the response patterns of alpine meadows to exogenous N input, which was mainly caused by decreased soil inorganic N under warming. The estimation of N thresholds highlights that alpine meadows are more sensitive to future N deposition than other types of grasslands.

[Responses of normalized difference vegetation index (NDVI) to precipitation changes on the grassland of Tibetan Plateau from 2000 to 2015.] / 2000­2015年青藏高原草地归一化植被指数对降水变化的响应.

Precipitation change is an important factor in the inter-annual variation of grassland growth on the Tibetan Plateau. The total amount, distribution pattern and concentration time are three basic characteristics of precipitation change. The temporal and spatial characteristics of precipitation change were analyzed based on climate data of 145 meteorological stations on the Tibetan Plateau and nearby areas from 2000 to 2015. The total precipitation amount was characterized by annual precipitation, distribution pattern of precipitation during the year was characterized by improved precipitation concentration index (PCI), and precipitation centroid (PC) was defined to indicate the change in precipitation concentrated time. To better illustrate the response of grassland to precipitation change, vegetation growth status was characterized by the maximum value of normalized difference vegetation index (NDVImax). Results indicated that the annual precipitation and PCI had an apparent gradient across the whole plateau and the latest PC occurred in the southern plateau. NDVImax of alpine shrub grassland was significantly correlated with the change of PCI,increased with even distribution of precipitation during growth period, and limited by the total annual precipitation. Alpine meadow did not show significantly correlations with these three indices. The inter-annual variability of NDVImax of steppe was controlled by both PCI and PC. NDVImaxof alpine desert grassland was mainly controlled by annual precipitation. In addition to annual total amount of precipitation, the distribution characteristics of precipitation should be further considered when the influence of precipitation change on different types of vegetation on the Qinghai Tibet Plateau was studied.

Enhanced Activities of δ Subunit-containing GABAA Receptors Blocked Spinal Long-term Potentiation and Attenuated Formalin-induced Spontaneous Pain.

The δ subunit-containing γ-Aminobutyric acid type A receptors (δ-GABAARs) are located at extrasynaptic sites and persistently active in the control of neuronal excitability. Here we recorded primary afferent C fiber-evoked field potentials in the superficial dorsal horn of rat spinal cords in vivo and investigated the possible influence of δ-GABAARs activities on nociceptive synaptic transmission. We found that δ-GABAARs-preferring agonist 4,5,6,7-tetrahydroisoxazolol [4,5-c] pyridine-3-ol (THIP), when topically applied onto spinal cord dorsum, inhibited the basal synaptic responses in a dose-dependent manner. Low-frequency stimulation (LFS) of sciatic nerves elicited long-term potentiation (LTP) of C fiber transmission, a synaptic correlate of central sensitization. Pretreatment with THIP before LFS delivery blocked the induction of LTP. When applied at 30 min and 180 min post-LFS, THIP reduced the magnitudes of established LTP. Intraplantar injection of formalin naturally evoked LTP in anesthetized rats. Spinal administration of THIP not only reversed formalin-induced LTP, but alleviated the spontaneous painful behaviors and mechanical hyperalgesia. Biochemical analysis demonstrated that δ-GABAARs activation by THIP decreased the synaptic expression and phosphorylation of AMPA receptor GluA1 subunit in formalin-injected rats, and meanwhile, increased synaptic GluA2 content, allowing the switch of GluA2-lacking AMPA receptors to GluA2-containing ones at synapses. THIP also suppressed the synaptic accumulation and phosphorylation of NMDA receptor GluN1 subunit in formalin-injected rats. Our data suggested that enhanced δ-GABAARs activities blunted the initiation and maintenance of spinal LTP, which correlated with the amelioration of central sensitization of nociceptive behaviors.

[Responses of plant community structure and species composition to warming and N addition in an alpine meadow, northern Tibetan Plateau, China]. / 藏北高寒草甸群落结构与物种组成对增温与施氮的响应.

Global climate warming and increasing nitrogen (N) deposition, as controversial global environmental issues, may distinctly affect the functions and processes of terrestrial ecosystems. It has been reported that the Qinghai-Tibet Plateau has been experiencing significant warming in recent decades, especially in winter. Previous studies have mainly focused on the effects of warming all the year round; however, few studies have tested the effects of winter warming. To investigate the effects of winter warming and N addition on plant community structure and species composition of alpine meadow, long-term N addition and simulated warming experiment was conducted in alpine meadow from 2010 in Damxung, northern Tibet. The experiment consisted of three warming patterns: Year-round warming (YW), winter warming (WW) and control (NW), crossed respectively with five N gradients: 0, 10, 20, 40, 80 kg N·hm-2·a-1. From 2012 to 2014, both warming and N addition significantly affected the total coverage of plant community. Specifically, YW significantly decreased the total coverage of plant community. Without N addition, WW remarkably reduced the vegetation coverage. However, with N addition, the total vegetation coverage gradually increased with the increase of N level. Warming and N addition had different effects on plants from different functional groups. Warming significantly reduced the plant coverage of grasses and sedges, while N addition significantly enhanced the plant coverage of grasses. Regression analyses showed that the total coverage of plant community was positively related to soil water content in vigorous growth stages, indicating that the decrease in soil water content resulted from warming during dry seasons might be the main reason for the decline of total community coverage. As soil moisture in semi-arid alpine meadow is mainly regulated by rainfalls, our results indicated that changes in spatial and temporal patterns of rainfalls under the future climate change scenarios would dramatically influence the vegetation coverage and species composition. Additionally, the effects of increasing atmospheric N deposition on vegetation community might also depend on the change of rainfall patterns.

Fever as a first manifestation of advanced gastric adenosquamous carcinoma: a case report.

Gastric adenosquamous carcinoma (ASC) is a rare type of gastric cancer. It is a mixed neoplasm, consisting of glandular cells and squamous cells. It is often diagnosed at an advanced stage, thus carrying a poor prognosis. We describe a case of a 73-year-old male, who presented with refractory fever and an intra-abdominal mass on imaging. He underwent a laparoscopic exploration followed by a successful totally laparoscopic total gastrectomy with D2 lymphadenectomy for gastric cancer. Postoperative pathology revealed primary gastric ASC (T4aN0M0). The patient received adjuvant radiotherapy and chemotherapy with S1 and is alive 20 mo after surgery without recurrence. This is the first case of advanced gastric ASC with fever as the initial presentation treated with totally laparoscopic total gastrectomy reported in the English literature.

[Effects of nitrogen and phosphorous fertilization on community structure and productivity of degraded alpine meadows in northern Tibet, China].

Abstract: Fertilization is an effective management measure for recovery of degraded grasslands. To better understand the effects of fertilization on community structure and productivity of lightly and severely degraded alpine meadows, we conducted a fertilization experiment in northern Tibet since 2008. The treatments were addition of nitrogen (N) alone (50 kg N x hm(-2) x a(-1), LN; 100 kg N x hm(-2) x a(-1), HN) or addition of both phosphorus (P) and N (50 kg N x hm(-2) x a(-1) +50 kg P x hm(-2) x a(-1), LN+P; 100 kg N x hm(-2) x a(-1) +50 kg P x hm(-2) x a(-1), HN+P) in each of the two types of degraded alpine meadows. N addition alone significantly affected plant community coverage or productivity in neither the slightly nor the severely degraded alpine meadow, while addition of both N and P significantly increased plant community coverage, aboveground and below- ground biomass of the alpine meadows. This suggested that productivity of this alpine meadow is co-limited by N and P. HN and HN+P significantly decreased species richness and evenness in the lightly degraded grassland, indicating that HN was not beneficial for the lightly degraded grassland to maintain species diversity and community stability. N addition significantly reduced the root to shoot ratio in the severely degraded meadow. In the lightly degraded meadow, N addition alone, especially with a high amount (HN) , enhanced the importance values (IV) and biomass of grasses, while fertilization with both N and P increased those of sedges. In the severely degraded meadow, fertilization had little effect on IV of grasses or sedges, but improved biomass of forbs. The results suggested that LN+P could be employed in recovery of lightly degraded alpine meadows, but other management measures such as fencing and reseeding may be needed for recovery of severely degraded alpine meadows.

[Analysis on vegetations spectral characteristics along the altitudinal gradients in south-facing slope of Dangxiong Valley].

The present study focused on variation of vegetation types and canopy spectra along the altitudinal gradients in south-facing slope of Dangxiong valley in Tibet. Spectral extraction methods including red edge analysis and vegetation indices were used for vegetation spectral characteristics analysis. Through the hierarchical clustering analysis based on the vegetation spectral features, the feasibility of remote sensing classification of vegetation types along the elevation gradients in the experimental area was evaluated. The experimental results showed that: there were significant differences in spectral features including water index (WI), red edge POSITION (REP), and normalized difference vegetation index (NDVI) in different plots along elevation gradients in the study area, and there were strong correlations between WI and leaf water content, REP and dry biomass, NDVI and vegetation coverage. The hierarchical clustering analysis result of 12 vegetation samples along the altitudinal gradients is consistent with the ground survey, which shows that the selected vegetation spectral features can characterize the vertical distribution of vegetation types in the experimental area. The vegetation spectral analysis in this study can provide the priori knowledge support of spectral characteristics for the vegetation vertical distribution information extraction in the Tibet Plateau.

Primary adenosquamous carcinoma of the jejunum.

Adenosquamous carcinomas of the small intestine are extremely rare, with only three documented jejunal and three ileal cases being reported in the English-language medical literature. Presented herein is a case of primary jejunal adenosquamous carcinoma in an 80-year-old woman. The jejunal carcinoma consisted predominantly of a squamous component throughout the tumor but peritoneal nodules carrying metastases from the adenocarcinoma element were noted, making it the first case of jejunal adenosquamous carcinoma with metastases from the adenocarcinoma component. The finding that metastases could arise from the minor component of a jejunal adenosquamous carcinoma indicates that an accurate diagnosis must be based upon thorough examination of both the primary and the metastases, not just mesenteric nodule biopsy alone. Histological foci of closely intermingled squamous and glandular components with apparent morphological transition were noted, indicating the pathogenetic possibility that the squamous component might arise by transformation from the glandular element. The squamous component was strongly positive with immunostaining for p63 (nuclear staining) and for cytokeratin 10/13 (cytoplasmic staining), while the adenocarcinoma element was negative. The immunohistochemical results suggest that p63 and cytokeratin 10/13 might be useful in identifying squamous differentiation in jejunal carcinoma.

[Ecological water requirement of forests in Loess Plateau].

Converting degraded farmlands to forest or grass lands is the best approach to reduce the soil erosion, the Loess Plateau is faced with most serious ecological disaster. Loess Plateau located in the arid and semi-arid regions is a fragile region with the characteristics of little precipitation and intensive evapotranspiration. Therefore, water is the most important factor limiting the eco-restoration and construction of the vegetation in the region. According to the latest digital land use map and with GIS, the ecological water requirement for forests in Loess Plateau was estimated, and by the water balance of the forest ecosystem in their growing season, the ecological water shortage was calculated. The results revealed that the minimum and suitable ecological water requirement of the forests in Loess Plateau were approximately 262.49 x 10(8) m3 and 421.34 x 10(8) m3 respectively. Accordingly, if it is taken the minimum ecological water requirement as the quota, the area of forests suffered water shortage was about 7,639.09 km2, 9.1% of all, and the ecological water shortage was amounted to 4.77 x 10(8) m3; if it is taken the suitable ecological water requirement as the quota, the forest land area suffered water shortage was about 57.7% of all, and the ecological water shortage was roughly 58.55 x 10(8) m3.