The gastrointestinal tract is the largest digestive organ and the largest immune organ and detoxification organ, which is vital to the health of the body. Drosophila is a classic model organism, and its gut is highly similar to mammalian gut in terms of cell composition and genetic regulation, therefore can be used as a good model for studying gut development. target of rapmaycin complex 1 (TORC1) is a key factor regulating cellular metabolism. Nprl2 inhibits TORC1 activity by reducing Rag GTPase activity. Previous studies have found that nprl2 mutated Drosophila showed aging-related phenotypes such as enlarged foregastric and reduced lifespan, which were caused by over-activation of TORC1. In order to explore the role of Rag GTPase in the developmental defects of the gut of nprl2 mutated Drosophila, we used genetic hybridization combined with immunofluorescence to study the intestinal morphology and intestinal cell composition of RagA knockdown and nprl2 mutated Drosophila. The results showed that RagA knockdown alone could induce intestinal thickening and forestomach enlargement, suggesting that RagA also plays an important role in intestinal development. Knockdown of RagA rescued the phenotype of intestinal thinning and decreased secretory cells in nprl2 mutants, suggesting that Nprl2 may regulate the differentiation and morphology of intestinal cells by acting on RagA. Knockdown of RagA did not rescue the enlarged forestomach phenotype in nprl2 mutants, suggesting that Nprl2 may regulate forestomach development and intestinal digestive function through a mechanism independent of Rag GTPase.
Animals ; Drosophila/genetics* ; Mechanistic Target of Rapamycin Complex 1/metabolism* ; Mammals/metabolism* ; Carrier Proteins ; Tumor Suppressor Proteins/metabolism* ; Drosophila Proteins/genetics*

To study the molecular mechanism of salt stress response of peanut small GTP binding protein gene AhRabG3f, a 1 914 bp promoter fragment upstream of the start codon of AhRabG3f gene (3f-P) from peanut was cloned. Subsequently, five truncated fragments (3f-P1-3f-P5) with lengths of 1 729, 1 379, 666, 510 and 179 bp were obtained through deletion at the 5' end, respectively. Plant expression vectors where these six promoter fragments were fused with the gus gene were constructed and transformed into tobacco by Agrobacterium-mediated method, respectively. GUS expression in transgenic tobacco and activity analysis were conducted. The gus gene expression can be detected in the transgenic tobacco harboring each promoter segment, among which the driving activity of the full-length promoter 3f-P was the weakest, while the driving activity of the promoter segment 3f-P3 was the strongest. Upon exposure of the transgenic tobacco to salt stress, the GUS activity driven by 3f-P, 3f-P1, 3f-P2 and 3f-P3 was 3.3, 1.2, 1.9 and 1.2 times compared to that of the transgenic plants without salt treatment. This suggests that the AhRabG3f promoter was salt-inducible and there might be positive regulatory elements between 3f-P and 3f-P3 in response to salt stress. The results of GUS activity driven by promoter fragments after salt treatment showed that elements included MYB and GT1 between 1 930 bp and 1 745 bp. Moreover, a TC-rich repeat between 682 bp and 526 bp might be positive cis-elements responsible for salt stress, and an MYC element between 1 395 bp and 682 bp might be a negative cis-element responsible for salt stress. This study may facilitate using the induced promoter to regulate the salt resistance of peanut.
Arachis/genetics* ; Fabaceae/genetics* ; GTP-Binding Proteins/metabolism* ; Gene Expression Regulation, Plant ; Glucuronidase/metabolism* ; Plant Proteins/metabolism* ; Plants, Genetically Modified/genetics* ; Salt Stress ; Stress, Physiological/genetics* ; Tobacco/genetics*

Objective:To prepare urokinase targeted thrombus microbubbles combined with low frequency ultrasound to dissolve thrombus in rabbit femoral artery, and to explore the mechanism of thrombolysis through the change of urokinase concentration.Methods:Twenty-four rabbits with thrombosed femoral artery were randomly divided into four treatment groups: urokinase alone (UK) group, ultrasound with non-targeted microbubble and urokinase (US+ M+ UK) group, platelet-targeted microbubble with urokinase (R+ UK) group, ultrasound with platelet-targeted microbubble and urokinase (US+ R+ UK) group. Thrombus-targeted microbubbles were injected through the ear vein and irradiated by ultrasound for 30 minutes, and the pulsed Doppler blood flow meter was used to continuously monitor the blood flow at 0 min, 10 min, 20 min, 30 min, 60 min, 90 min and 120 min after injection. At the same time, the changes in urokinase concentration were monitored, and the characteristics of blood flow and urokinase concentration changes were analyzed.Results:UK and US+ M+ UK groups failed to show recanalization at 120 minutes after treatment.In contrast, the US+ R+ UK and R+ UK groups both achieved persistent recanalization( P<0.001). Compared with the basal state. Both the R+ UK group and R+ UK+ US group showed a decrease at 60 minutes, and the difference was statistically significant( P<0.05). Conclusions:In the thrombolysis process of low-frequency ultrasound combined with a targeted contrast agent carrying urokinase, the targeting can make the local urokinase concentration of thrombus reach the highest level. At the same time, ultrasound and targeted microbubbles promote urokinase to enter the thrombus, and finally achieve the strongest thrombolytic effect.

Objective? To explore the application effects of the nursing team led multidisciplinary cooperative intervention in postoperative cognitive rehabilitation of patients with ischemic stroke. Methods? From May 2016 to June 2017,80 patients with cognitive dysfunction after ischemic stroke in a ClassⅢ Grade A hospital in Beijing were selected and randomly divided into observation group and control group based on random number table,with 40 patients in each group. The control group received the routine nursing care and on the basis of that, the observation group received the nursing team led multidisciplinary cooperative intervention. The cognitive function and quality of life in two groups were assessed by the Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and World Health Organization Quality of Life in Brief Version (WHOQOL-BRIEF) before and 6 months after intervention. Results? Six months after intervention,the MMSE and MoCA scores of the observation group were (29.35±0.58) and (26.70±2.03) scores, which were statistically different from the control group (t=5.64, 4.15;P＜0.05). The total score,scores of physiological field,psychological field and social field in the quality of life of the observation group were (64.59±5.14), (16.94±1.43),(17.10±1.55), and (15.60±1.07)scores,which were significantly higher than the control group (t=4.82, 5.60,7.41,3.23;P＜0.01). Conclusions? The nursing team led multidisciplinary cooperative intervention can improve the cognitive function and quality of life in the patients after ischemic stroke and accelerate postoperative recovery forming a virtuous circle.

Objective To explore the express of von willebrand factor(vWF) and tissue factor(TF) in ultrasound combined with urokinase and RGDS-targeted microbubbles for recanalization of occluded rabbit femoral artery.Methods A total of 42 rabbits with platelet-rich thrombi in the femoral artery were randomized into 7 treatment groups (n =6 in each group):①ultrasound alone (US) group;②ultrasound,non-targeted microbubbles (US + M) group;③urokinase alone (UK) group;④ ultrasound,non-targeted microbubble and urokinase (US + M + UK) group;⑤ ultrasound,platelet-targeted microbubble (US + R) group;⑥ platelet-targeted microbubble,urokinase (R + UK) group;⑦ ultrasound,platelet-targeted microbubble and urokinase (US + R + UK) group.A total of 6 ml of infusion liquor of urokinase,RGDS and microbubbles (SonoVue) were mixed by the direct conjugation method,infusion via vein within 20 min.Ultrasound was conducted to lyse the clot for 30 min.The recanalization and the velocity tracing change of blood flow in thrombolytic process and the express of vWF and TF were evaluated at 120 min post treatment.Results For US,UK,US + M,US + R and US + M + UK groups,recanalization were failed.The R + UK and US + R + UK groups were partly recanalizated or completed recanalized (P ＜0.001).vWF and TF were positive in US,US + M,US + R and R + UK group.vWF and TF were negative in US + M + UK,UK and US + R + UK groups.Conclusions The combined effects of low frequency ultrasound and targeted microbubble combined with urokinase can inhibit the expression of vWF and TF,and then promote the thrombolysis,however it can affect the expression of vWF and TF after reperfusion.

Objective To explore the correlation between matrix metalloproteinase‐9(MMP‐9) and cardiac function ,aneu‐rysm volume in the process of left ventricular aneurysm (LVA) formation .Methods Rabbit models of LVA were established in 20 New Zealand rabbits by lighting left anterior descending artery and left circumflex artery .Two‐dimensional echocardiography were performed at preoperative and postoperative 1 d ,2 d ,3 d ,1 w ,2 w ,3 w ,4 w .The survived animals which had LVA formed were en‐rolled .Real time three‐dimensional echocardiography was performed to obtain left ventricular volume ,LVEF and LVA volume ,The MMP‐9 serum concentration was measured by ELISA .Results (1)There were a significant increase in the serum concentration of MMP‐9 from postoperative 1 d and arrived at the peak at postoperative 3 d ,there were higher than preoperative at postoperative 4 w (P<0 .05) .(2)Compared with preoperative ,the LVEDV ,LVESV and LVAV volume at postoperative showed an increase trend (P<0 .05) ,while there was a decreasing trend in LVEF(P<0 .05) .(3)The MMP‐9 had the better relationship with LVEF at post‐operative 1 d ,2 d ,3 d ,1 w ,2 w ,3 w ,4 w ,in which MMP‐9 had the tightest relationship with LVEF at postoperative 3 d (r=0 .731 , P<0 .05) .MMP‐9 had the better relationship with LVA volume at postoperative 2 d ,3 d ,1 w ,2 w ,3 w ,4 w ,in which MMP‐9 had the tightest relationship with LVA volume at postoperative 3 d (r=0 .636 ,P<0 .05) .Conclusion The MMP‐9 had an effect on cardiac function and LVA volume in the process of LVA formation .

OBJECTIVE:To observe therapeutic efficacy and safety of Yinxing damo injection for neural function recovery af-ter hypertensive intracerebral hemorrhage (HICH) minimally invasive surgery. METHODS:84 HICH patients were randomly di-vided into control group and observation group with 42 patients in each group. Both groups received CT guiding minimally inva-sive aspiration. Control group was given western medicine baseline therapy,such as dehydration and intracranial decompression, controlling blood pressure,preventing infection,alimenting never,symptomatic treatment. Observation group was additionally giv-en Yinxing damo injection 20 ml,ivgtt,bid. Treatment course lasted for 14 d. NIHSS score,GCS score and Fugl-Meyer motor function assessment scale score were comducted before and after treatment. The serum levels of NSE,serum C3,C4 and hs-CRP were determined in 2 groups before and after treatment. RESULTS:After treatment,the effective rate of observation group (85.71%)was better than that of control group(66.67%),with statistical significance(P<0.05);after treatment,NIHSS score, Fugl-Meyer score and GCS score of 2 groups were all better than before,the observation group was better than the control group, the levels of C3 and C4 in observation group were lower than in control group,with statistical significance(P<0.01). There was one case of allergic reaction that the patient can tolerate. Magnesium sulfate for external use was given,which did not affect the treatment. CONCLUSIONS:Yinxing damo injection could improve neurologic impairment,promote the recovery of patients and have good safety.

Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78％ ± 0.52％ vs.0.47％ ± 0.15％,P＜ 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P＜ 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P＜ 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P＜ 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P＜ 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P ＜ 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P ＜ 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P ＜ 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.

Objective To study the change of thrombosis in rabbits arteries by electron microscope after thromb was lyses by urokinase targeted microbubbles.Methods A total of 24 rabbits with plateletrich thrombi in the femoral artery were randomized into 4 treatment groups (n = 6) :(1) ultrasound alone (US) ; (2) urokinase alone (UK); (3) ultrasound, non-targeted microbubble and urokinase (US + M + UK) ;(4) ultrasound, platelet-targeted microbubble and urokinase (US + R + UK).Urokinase targeted microbubbles were in conjunction with the surface of commercial microbubbles (SonoVue) by direct conjugation method.US was simultaneously applied transcutaneously over the thrombus up to 30 min.The thrombolytic effect was based on ultrasound, blood flow and histological observations at 120 min post treatment.Results US + R + UK group was completed recanalization (P ＜ 0.001).Scanning electron microscope examination showed thrombosis of the fiber network structure damage.Conclusions The main electron microscope change of RGDS urokinase targeted microbubbles dissolve thromb is thrombus fibrin network structure damage and fibrin dissolution.

Objective To determine the effect of the combined use of urokinase and glycoprotein Ⅱb/Ⅲa-targeted microbubbles prepared by direct conjugation method to dissolve the thromb in vivo and analyse the velocity tracing change of blood flow and explore the possible mechanism. Methods A total of 42 rabbits with platelet-rich thrombi in the femoral artery were randomized into 7 treatment groups ( n = 6 in each group): 1) ultrasound alone (US); 2) ultrasound plus non-targeted microbubbles ( US + M); 3) urokinase alone (UK) ;4) ultrasound, non-targeted microbubble and urokinase (US + M + UK); 5) ultrasound plus platelet-targeted microbubble ( US + R); 6) platelet-targeted microbubble plus urokinase (R + UK); 7)ultrasound, platelet-targeted microbubble and urokinase (US + R + UK). A total of 6 ml of infusion liquor of Urokinase,RGDS and microbubbles (SonoVue) were mixed by 1 ∶ 1 ∶ 1 ratio by the direct conjugation method, infusion via vein within 20 min. Ultrasound was conducted to lyse the clot for 30 min. The recanalization and the velocity tracing change of blood flow in thrombolytic process were evaluated at 120 min post treatment. Results For US, UK, US + M, US + R and US + M + UK groups, recanalization was failed. The R + UK and US + R + UK was recanalizated ( P ＜0.001 ). The blood flow velocity tracing was small and low width in US,UK, US + M, US + R and US + M + UK groups. The wave was high width and disorderly under the thrombolysis therapy in the R + UK and US + R + UK. The thrombolytic effect was demonstrated by the high-width and disorderly resonance changes in the blood flow spectrum during the thrombolytic therapy of US + R + UK. Conclusions The blood flow spectrum of groups had different characteristics in vivo when thrombus was issolved,ultrasonic resonance might be the possible mechanism.