Distinct Circuits From the Central Lateral Amygdala to the Ventral Part of the Bed Nucleus of Stria Terminalis Regulate Different Fear Memory.

BACKGROUND: The ability to differentiate stimuli that predict fear is critical for survival; however, the underlying molecular and circuit mechanisms remain poorly understood. METHODS: We combined transgenic mice, in vivo transsynaptic circuit-dissecting anatomical approaches, optogenetics, pharmacological methods, and electrophysiological recording to investigate the involvement of specific extended amygdala circuits in different fear memory. RESULTS: We identified the projections from central lateral amygdala (CeL) protein kinase C δ (PKCδ)-positive neurons and somatostatin (SST)-positive neurons to GABAergic (gamma-aminobutyric acidergic) and glutamatergic neurons in the ventral part of the bed nucleus of stria terminalis (vBNST). Prolonged optogenetic activation or inhibition of the PKCδCeL-vBNST pathway specifically reduced context fear memory, whereas the SSTCeL-vBNST pathway mainly reduced tone fear memory. Intriguingly, optogenetic manipulation of vBNST neurons that received the projection from PKCδCeL neurons exerted bidirectional regulation of context fear, whereas manipulation of vBNST neurons that received the projection from SSTCeL neurons could bidirectionally regulate both context and tone fear memory. We subsequently demonstrated the presence of δ and κ opioid receptor protein expression within the CeL-vBNST circuits, potentially accounting for the discrepancy between prolonged activation of GABAergic circuits and inhibition of downstream vBNST neurons. Finally, administration of an opioid receptor antagonist cocktail on the PKCδCeL-vBNST or SSTCeL-vBNST pathway successfully restored context or tone fear memory reduction induced by prolonged activation of the circuits. CONCLUSIONS: Together, these findings establish a functional role for distinct CeL-vBNST circuits in the differential regulation and appropriate maintenance of fear.

A Deep Mesencephalic Nucleus Circuit Regulates Licking Behavior.

Licking behavior is important for water intake. The deep mesencephalic nucleus (DpMe) has been implicated in instinctive behaviors. However, whether the DpMe is involved in licking behavior and the precise neural circuit behind this behavior remains unknown. Here, we found that the activity of the DpMe decreased during water intake. Inhibition of vesicular glutamate transporter 2-positive (VGLUT2+) neurons in the DpMe resulted in increased water intake. Somatostatin-expressing (SST+), but not protein kinase C-δ-expressing (PKC-δ+), GABAergic neurons in the central amygdala (CeA) preferentially innervated DpMe VGLUT2+ neurons. The SST+ neurons in the CeA projecting to the DpMe were activated at the onset of licking behavior. Activation of these CeA SST+ GABAergic neurons, but not PKC-δ+ GABAergic neurons, projecting to the DpMe was sufficient to induce licking behavior and promote water intake. These findings redefine the roles of the DpMe and reveal a novel CeASST-DpMeVGLUT2 circuit that regulates licking behavior and promotes water intake.

The Effect of Continuous Passive Motion in Patients Treated With Total Knee Arthroplasty for Osteoarthritis: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

OBJECTIVE: The aim of the study was to identify the effectiveness of the continuous passive motion application on clinical outcomes after total knee arthroplasty, based on evidence from recently published high-quality randomized controlled trials. DESIGN: Two reviewers retrieved platforms of PubMed, Embase, and CENTRAL independently, for identifying eligible randomized controlled trials evaluating the effect of continuous passive motion applied after total knee arthroplasty for knee osteoarthritis. Subgroup meta-analyses were performed for all syntheses based on the follow-up intervals. RESULTS: A total of 10 randomized controlled trials, involving 841 patients, were finally included. Data were available for 15 different outcomes (including active/passive knee extension/flexion/full range of motion, Western Ontario and McMaster Universities Osteoarthritis Index-pain/physical function/stiffness/total score, visual analogue scale, time up and go, knee girth, Knee Society Scale-function/knee score), at several time points. In general, most of the pools demonstrated similar outcome between continuous passive motion and noncontinuous passive motion groups. Exclusively, the active knee extension at 1 wk (mean difference = 3.00, 95% confidence interval = 0.5-5.5, P = 0.019*), passive knee extension at 1 wk (mean difference = 3.00, 95% confidence interval = 0.28-5.72, P = 0.031*), and 3 mos (mean difference = 3.00, 95% confidence interval = 0.5-5.5, P = 0.019*) were shown to be significantly slightly different between two groups. CONCLUSIONS: This study demonstrated a limited role of continuous passive motion in patients operated with total knee arthroplasty. Thus, there is at this stage no indication for continuous passive motion procedures in patients operated with total knee arthroplasty as a standard postoperative care.

[Effects of fertilization and planting patterns on soil aggregate and carbon distribution in farmland of the Loess Plateau, Northwest China]. / ç§æ¤æ¨¡å¼å’Œæ–½è‚¥å¯¹é»„åœŸæ—±å¡¬å†œç”°åœŸå£¤å›¢èšä½“åŠå ¶ç¢³åˆ†å¸ƒçš„å½±å“.

Based on a long-term experiment in the Changwu Agro-ecological Experimental Station in Xianyang, Shaanxi, China, we examined the effects of fertilization and planting patterns on soil aggregate quantity, aggregate stability and total carbon and organic carbon distribution in different aggregate fractions through dry and wet sieving methods, as well as the TOC combustion method. There were ten treatments, including uncultivated (R), wheat continuous cropping (CK/W), wheat-corn rotation (L), and nitrogen fertilizer (N), phosphorus fertilizer (P), nitrogen and phosphorus fertilizer (NP), organic fertilizer (M), nitrogen and organic fertilizer (NM), phosphorus and organic fertilizer (PM), nitrogen and phosphorus and organic fertilizer (NPM) for CK/W. The results showed that fertilizer application and planting patterns affected soil aggregate distribution and stability, the contents and contribution rates of total C and organic C. Force-stable aggregate was mainly constituted by >0.25 mm aggregate (>67%), which was reduced by fertilization. Continuous cropping decreased micro-aggregate while rotation facilitated it and the effect was larger than fertilization. Water-stable aggregate was mainly comprised of micro-aggregate (<0.25 mm), the contribution of which was larger than 61%. Both fertilizer application and planting pattern reduced water-stable micro-aggregate. Fertilizer application and planting pattern decreased the percentage of aggregate destruction rate (PAD) and increased macro-aggregate (>0.25 mm, R0.25) content. Organic fertilizer significantly improved total C and organic C concentrations in all the fractions of force-stable aggregates. Continuous cropping and rotation cropping increased total C concentration in all the aggregate fractions while rotation cropping significantly decreased organic C concentration. Single N and P fertilization decreased soil total C concentration, while mixed application of N and P fertilizers, and organic fertilizer significantly increased soil total C concentration. The effect of planting patterns on soil total C was lower than that of fertilization. Both continuous cropping and rotation cropping increased soil total C. Mixed application of N and P fertilizers, and organic fertilizer signifi-cantly increased soil organic C concentration while single N and P fertilization decreased it. The effect of planting patterns on soil organic C was lower than that of fertilization, while rotation cropping did not facilitate soil organic C. Micro-aggregate was the most notable size fraction to total carbon and organic C, with the contribution being 21.2%-33.6%. Fertilization and planting pattern increased the contribution rate of micro-aggregate in soil total C. NP and NPM significantly increased the contribution rate of micro-aggregate in soil total C and soil organic C. The effect of rotation cropping was most obvious in driving the contribution rate of micro-aggregate in soil total C and soil organic C.

[Effects of long-term fertilization on soil organic phosphorus fractions and wheat yield in farmland of Loess Plateau]. / 长期施肥对黄土旱塬农田土壤有机磷组分及小麦产量的影响.

Understanding the effects of long-term fertilization on soil organic phosphorus fractions and wheat yield in the Loess Plateau can provide theoretical support for improving phosphorus conversion, utilization, and rational use of fertilizer. We examined the effects of different fertilizer treatments on soil organic phosphorus fractions, wheat yield and soil properties of a farmland in the long-term (1984-2016) positioning test station of Changwu loess soil. There were eight treatments, including no fertilizer (CK), single application of nitrogen fertilizer (N), single application of phosphorus fertilizer (P), application of nitrogen and phosphorus fertilizer (NP), single application of organic fertilizer (M), nitrogen combined with organic fertilizer (MN), phosphorus combined with organic fertilizer (MP), nitrogen and phosphorus combined with organic fertilizer (MNP). The results showed that the range of soil organic phosphorus content was 244.7-429.1 mg·kg-1 after long-term fertilization. Except for the N treatment, organic phosphorus content was significantly increased by 15.4%-47.9% compared to CK. Long-term application of phosphorus fertilizer changed the content of organic phosphorus fractions in the surface soil (0-20 cm). The treatments of MP and MNP significantly increased the contents of labile organic phosphorus and moderately labile organic phosphorus. The treatments of N, P and NP significantly reduced the content of moderately stable organic phosphorus. The treatments of N, P, NP, MN, MP, MNP all significantly increased the highly stable organic phosphorus. The ratio of soil organic phosphorus fractions to total organic phosphorus content was in order of moderately labile organic phosphorus > highly stable organic phosphorus > labile organic phosphorus > moderately stable organic phosphorus. After long-term fertilizer application, the combination of nitrogen and phosphorus fertilizers, especially with organic fertilizers, significantly increased wheat biomass yield and grain yield. Among all the examined soil properties, organic matter, Olsen-P and total inorganic phosphorus were significantly positively correlated with wheat yield. MP and M could significantly increase the content of Olsen-P, total phosphorus, total inorganic phosphorus, labile organic phosphorus and moderately labile organic phosphorus in the loess soil of Loess Plateau. Our results indicated that the organic and phosphorus fertilizers could improve soil phosphorus components that could be more easily absorbed by crops. In summary, the combination of nitrogen and phosphorus fertilizers, especially with organic fertilizers, could increase soil phosphorus supply in the region and promote the wheat yield, which is important for improving soil quality in the Loess Plateau.

[Effects of long-term fertilization on soil microbial abundance in farmland of the Loess Plateau, China]. / 长期施肥对黄土旱塬农田土壤微生物丰度的影响.

Based on a long-term experiment in the Changwu Agro-ecological Experimental Station, the real-time PCR analysis was used to examine the soil microbial abundance and to reveal the effects on soil microbial community under different long-term fertilization systems. The results showed that compared to the CK (barren field), the population of bacteria increased by 21% and archaea by 32% in treatment with inorganic fertilizer application. The abundance of bacteria in the treatment of chemical fertilizer combined with organic fertilizer increased by 37% and archaea by 36%. The treatment with chemical fertilizer mixed with organic fertilizer significantly increased the abundance of bacteria and archaea. The soil AOB increased by 7.13 times while the soil AOA only by 0.2 folds after 30-year application of chemical nitrogen fertilizer. AOB was highly responsive to fertilizer application, especially to nitrogen fertilizer. Compared with the single nitrogen application and the application of nitrogen fertilizer mixed with organic fertilizer, phosphorus fertilizer significantly increased the abundance of nifH and pmoA. The content of nifH, nirS cd and pmoA in the abandoned land was significantly higher than that in the cultivated soil. Results from the correlation analysis on soil basic physical and chemical properties indicated that soil pH, total nitrogen and organic carbon were key factors affecting soil microbial community abundance. In conclusion, long-term fertilization significantly changed soil microbial abundance, and fertilization patterns and cultivating methods had significant effect on microbial community abundance.

Rostral and Caudal Ventral Tegmental Area GABAergic Inputs to Different Dorsal Raphe Neurons Participate in Opioid Dependence.

Both the ventral tegmental area (VTA) and dorsal raphe nucleus (DRN) are involved in affective control and reward-related behaviors. Moreover, the neuronal activities of the VTA and DRN are modulated by opioids. However, the precise circuits from the VTA to DRN and how opioids modulate these circuits remain unknown. Here, we found that neurons projecting from the VTA to DRN are primarily GABAergic. Rostral VTA (rVTA) GABAergic neurons preferentially innervate DRN GABAergic neurons, thus disinhibiting DRN serotonergic neurons. Optogenetic activation of this circuit induces aversion. In contrast, caudal VTA (cVTA) GABAergic neurons mainly target DRN serotonergic neurons, and activation of this circuit promotes reward. Importantly, µ-opioid receptors (MOPs) are selectively expressed at rVTA→DRN GABAergic synapses, and morphine depresses the synaptic transmission. Chronically elevating the activity of the rVTA→DRN pathway specifically interrupts morphine-induced conditioned place preference. This opioid-modulated inhibitory circuit may yield insights into morphine reward and dependence pathogenesis.

[Effects of fertilizing regime and planting age on soil calcium decline in Luochuan apple orchards]. / æ–½è‚¥æ–¹å¼å’Œå›­é¾„å¯¹æ´›å·è‹¹æžœå›­åœŸå£¤é’™ç´ é€€åŒ–çš„å½±å“.

This study was conducted to assess the effects of fertilizing regime and orchard planting age on soil calcium contents and stocks in the apple orchards on the Loess Plateau. The apple orchards in Luochuan County, one of the best regions for apple plantation in the world, were selec-ted in this study. The contents of calcium carbonate,water-soluble calcium and exchangeable cal-cium at 0-100 cm soil layer under different fertilizing regimes and various planting ages were mea-sured, their stocks were calculated and their variation features were analyzed. The results showed that soil in the apple orchards in the study region was characterized by the decline in calcium contents. The decline was more serious in apple orchards with long-term application of chemical fertili-zer than in those with combined application of chemical fertilizer and farmyard manure. The average contents of calcium carbonate, water-soluble calcium and exchangeable calcium at 0-100 cm soil layer in apple orchards with long-term application of chemical fertilizer decreased by 38.8%, 25.4% and 5.6% respectively than those in the apple orchards with long-term application of both chemical fertilizer and farmyard manure. The stocks of calcium carbonate, water-soluble calcium and exchangeable calcium decreased by 36.4%, 26.0% and 4.3%, respectively. The decline of soil cal-cium was aggravated with the increase of orchard planting age. The contents of calcium carbonate, water-soluble calcium and exchangeable calcium at 0-100 cm soil layer in orchards of more than 25 years of planting age decreased by 48.8%, 69.4% and 39.5% respectively, compared with orchards of less than 10 years of planting age, and the stocks decreased by 40.8%, 64.1% and 33.0%, respectively. These results indicated that either long-term application of chemical fertilizer or long-term plantation of apple trees obviously depleted soil calcium carbonate, water-soluble calcium and exchangeable calcium. Therefore, it was recommended that application of chemical fertilizer and farmyard manure should be combined to mitigate soil calcium decline, and calcium management should be strengthened in apple orchards of more than 25 years of planting age. The fertilizing regime was a driving factor of soil calcium decline which had a significant temporal (orchard planting age) and spatial (soil depth) effect.

[Construction of T vectors based on Xcm I recognition site and optimization of PCR fragments for ligation].

OBJECTIVE: To construct T vectors based on Xcm I recognition site and optimize the PCR fragments for its ligation. METHODS: We firstly cloned the human histone H4 cDNA containing one Xcm I recognition site at both its 5' and 3' end into pCDNA 3.0 vector and then generated T vector with pCDNA 3.0 backbone by cutting the recombinant plasmid with Xcm I. To increase the ligation efficiency, the primers were firstly phosphorylated before DNA fragments amplification and then the PCR products were treated with Taq DNA polymerase and dATP after PCR amplification. Two DNA fragments with the length of 312 bp and 1 329 bp were ligated to it and the ligation mixture was transformed into E. coli DH5α competent cells and the positive rates of the transformants were evaluated by PCR and DNA agarose gel electrophoresis. RESULTS: Our results showed that the T vector produced by our method could ligate to the target DNA fragments with high efficiency. Besides, the phosphorylation state of the primers used for PCR amplification is also an important factor determining the cloning efficiency. What's more, as for longer DNA fragments, retreatment with Taq DNA polymerase and dATP after PCR amplification and purification could improve the ligation efficiency significantly. CONCLUSION: Our protocol may overcome the dependence on blue/white screening to get positive clones and provide a potent way to generate T vectors and ligate them to the target PCR fragment.

Adaptive power projection method for accumulative EEG classification.

For the dynamic classification of motor imagery mind states in the brain-computer interface (BCI), we propose a power projection based feature extraction method to classify the electroencephalogram (EEG) signals by combining information accumulative posterior Bayesian approach. This method improves the classification accuracy by maximizing the average projection energy difference of the two types of signals. The experimental results on two BCI competition datasets show that the classification accuracy is about 90%. The results of the classification accuracy and mutual information demonstrate the effectiveness of this method.

[Promoting effect of hyperhomocysteinemia on vascular calcification in rats].

AIM: To explore the effect of hyperhomocysteinemia on vascular calcification and the underlying mechanism of it. METHODS: Arterial calcification of Sprague-Dawley rats was induced by vitamin D3 plus nicotine. Hyperhomocysteinemia was established by feeding high methionine diet for six weeks and was assessed b y plasma total homocysteine (tHcy) level detected by HPLC method. Calcification was confirmed by von Kossa staining, measurement of calcium content, alkaline phosphatases (ALP) activity and osteocalcin (OC) concentration of vascular tissue. Lipid conjugated dienes formation were determined to reflecting the production of lipid peroxide. RESULTS: The results showed that there were mass black granules deposited in aortic wall of the calcified rats by von Kossa staining. Calcium content, ALP activity, OC concentration in calcified rats increased by 8.09-fold, 45.57% and 2.81-fold compared with those of the control group (P < 0.01). Calcium content in calcified rats with high methionine diet increased by 34.29% compared with that of the calcified rats, while ALP activity and OC content decreased by 29.13% and 74.69% compared with that of the calcified rats. Lipid conjugated dienes formation in plasma of the rat with high methionine diet and of calcified rats with high methionine diet increased by 1.93 and 2.89-fold compared with those of the control group, respectively (P < 0.01), and in calcified rats with high methionine diet group was increased by 32.90% compared with that of high methionine diet group (P < 0.01). CONCLUSION: Hyperhomocysteinemia could promote vascular calcification, which might be mediated through the production of lipid peroxide.