Identification of the Interaction Between Gravin and {mu}-Opioid Receptors

The phosphorylation and desensitization of G-protein coupled receptors involve several protein kinases. Two of the major ones are protein kinase A (PKA) and protein kinase C (PKC). It has been reported that gravin, an A-kinase anchoring protein, plays a role in regulating p2-adrenergic receptor desensitization, resensitization, and internalization by recruiting both PKA and PKC, phosphatase, |3-arrestin, and clathrin to the receptor. Similarities between the regulation of p-opioid receptor and p2-adrenergic receptor suggest that a similar complex involving gravin may play a role in regulating p- opioid receptor signal transduction. To investigate this possibility, the current study examined the distribution of gravin, p-opioid receptor and p2-adrenergic receptor in SH- SY5Y cells as well as in AN3 CA cells cotrans fected with gravin and p-opioid receptor or p2-adrenergic receptor vector constructs. This study also examined the distribution of gravin, p-opioid receptor and p2-adrenergic receptor transgene in transfected AN3 CA cells after the agonist stimulation. Our data showed that gravin is distributed along the membrane of SH-SY5Y cells and partially colocalized with the p-opioid receptor and the p2-adrenergic receptor. The gravin transgene was also concentrated along the membrane of AN3 CA cells. Immunofluorescent microscopy showed extensive colocalization of gravin and either the p-opioid receptor transgene or the p2-adrenergic receptor transgene before agonist treatment. However, after agonist treatment, the p-opioid receptor and p2- adrenergic receptor transgenes translocated from plasma membrane to a perinuclear location, but there was no redistribution of grav in. The current study provided evidence that the recombinant gravin and the receptor proteins were expressed and distributed in a similar way as the native proteins. This validated their use in studying p-opioid receptor and p2-adrenergic receptor signal transduction and the role of gravin in regulating the function of these receptors. The current study also provided evidence that gravin might not interact with the receptors during the internalization of the receptors

Growth of tomato and cucumber seedlings under different light environments and their development after transplanting

Selecting suitable light conditions according to the plant growth characteristics is one of the important approaches to cultivating high-quality vegetable seedlings. To determine the more favorable LED light conditions for producing high-quality tomato and cucumber seedlings in plant factories with artificial light (PFALS), the growth characteristics of tomato and cucumber seedlings under seven LED light environments (CK, B, UV-A, FR, B+UV-A, UV-A+FR, and B+FR) and the development of these seedlings after transplanting into a plastic greenhouse were investigated. The results showed that the seedling height and hypocotyl length increased in treatments with far-red light supplementation (FR, UV-A+FR, and B+FR), but decreased in the B treatment, in both varieties. The seedling index of tomato seedlings increased in the B+UV-A treatment, while that of cucumber seedlings increased in the FR treatment. After transplanting into a plastic greenhouse, tomato plants that radiated with UV-A had greater flower numbers on the 15th day after transplanting. In cucumber plants of the FR treatment, the flowering time was significantly delayed, and the female flower exhibited at a lower node position. By using a comprehensive scoring analysis of all detected indicators, light environments with UV-A and FR were more beneficial for improving the overall quality of tomato and cucumber seedlings, respectively

The role and characteristics of low-level jet during a persistent rainstorm in Guangxi

In 2022, Guangxi experienced the strongest Dragon Boat Precipitation since the founding of the People's Republic of China. The persistent rainstorm process from 17 June to 22 June 2022 had large accumulations and overlapping falling areas, leading to floods, torrential floods, landslides, and other disasters. The impacts of the low-level jet and its characteristics during this process were analyzed based on multi-source observation data and ERA5 reanalysis data. The results show that: (1) The low-level jet enhanced significantly during the night, and the convective system developed on the north side of the jet core. During the daytime, the low-level jet weakened and the convective system gradually disappeared, resulting in the heavy precipitation being mainly concentrated during the night. (2) During the night, the positive vorticity zone on the left of the 850 hPa low-level jet coincided with the exit zone of the 925 hPa boundary-level jet in the northeast of Guangxi, which combined with the mountain terrain barrier, and caused deep low-level convergence. The low-level convergence was conducive to the enhancement of the upward movement in northeast Guangxi and favored the continuous development of the convective system. At the same time, the convective instability in the lower atmosphere also increased rapidly, which provided a favorable unstable stratification environment for heavy precipitation. (3) The variation of the low-level jet can be well explained by the inertial oscillation mechanism. During the day, the surface heating in the central and southern regions of Guangxi led to the gradual enhancement of turbulent friction, resulting in jet deceleration with the characteristics of sub-geostrophic. During the night, the turbulent friction was weakened, and the jet accelerated, which gradually presented the characteristics of super-geostrophic. The Coriolis force acting on ageostrophic wind was the main contributor to the momentum of the jet, while friction dissipation and vertical transport were the momentum sinks

Etiology and Clinical Characteristics of Influenza-Like Illness (ILI) in Outpatients in Beijing, June 2010 to May 2011

BACKGROUND: Since May 2009, exposure of the population of Beijing, China to pH1N1 has resulted in an increase in respiratory illnesses. Limited information is available on the etiology and clinical characteristics of the influenza-like illness (ILI) that ensued in adults following the pH1N1 pandemic. METHODS: Clinical and epidemiological data of ILI in adults was collected. A total of 279 throat swabs were tested for twelve respiratory viruses using multiplex RT-PCR. Clinical characteristics of influenza A in outpatients versus test-negative patients were compared using Pearson's χ2 and the Mann-Whitney U test. 190 swabs were tested for pH1N1 by virus isolation. Consultation rates for ILI were compared between 2009 and 2010. RESULTS: One or two virus were detected in 29% of the samples. Influenza A virus (FLU-A) accounted for 22.9% (64/279). Other viruses were present at a frequency less than 3.0%. Cough was significantly associated with Influenza A virus infection (χ2, p<0.001). The positive rate of FLU-A was consistent with changes in the ILI rate during the same period and there was a significant reduction in the incidence of ILI in 2010 when compared to 2009. During the 2010-2011 influenza season, the incidence peaked in January 2011 in Beijing and north China. CONCLUSIONS: Exposure to pH1N1 had no impact on typical influenza seasonal peaks, although FLU-A remained the predominant virus for 2010 in Beijing. Symptomatically, cough was associated with FLU-A infection. The positive rate of influenza virus was consistent with changes in the ILI rate during the same period and there was a significant reduction in the incidence of ILI in 2010 when compared to that of 2009

γδT Cells Suppress Liver Fibrosis via Strong Cytolysis and Enhanced NK Cell-Mediated Cytotoxicity Against Hepatic Stellate Cells

Liver fibrosis is the excessive accumulation of extracellular matrix proteins, resulting from maladaptive wound healing responses to chronic liver injury. γδT cells are important in chronic liver injury pathogenesis and subsequent liver fibrosis; however, their role and underlying mechanisms are not fully understood. The present study aims to assess whether γδT cells contribute to liver fibrosis regression. Using a carbon tetrachloride (CCl4)-induced murine model of liver fibrosis in wild-type (WT) and γδT cell deficient (TCRδ−/−) mice, we demonstrated that γδT cells protected against liver fibrosis and exhibited strong cytotoxicity against activated hepatic stellate cells (HSCs). Further study show that chronic liver inflammation promoted hepatic γδT cells to express NKp46, which contribute to the direct killing of activated HSCs by γδT cells. Moreover, we identified that an IFNγ-producing γδT cell subset (γδT1) cells exhibited stronger cytotoxicity against activated HSCs than the IL-17-producing subset (γδT17) cells upon chronic liver injury. In addition, γδT cells promoted the anti-fibrotic ability of conventional natural killer (cNK) cells and liver-resident NK (lrNK) cells by enhancing their cytotoxicity against activated HSCs. The cell crosstalk between γδT and NK cells was shown to depend partly on co-stimulatory receptor 4-1BB (CD137) engagement. In conclusion, our data confirmed the protective effects of γδT cells, especially the γδT1 subset, by directly killing activated HSCs and increasing NK cell-mediated cytotoxicity against activated HSCs in CCl4-induced liver fibrosis, which suggest valuable therapeutic targets to treat liver fibrosis

Electrophysiology as a prognostic indicator of visual recovery in diabetic patients undergoing cataract surgery

Purpose: Visual outcomes after cataract surgery in diabetic patients with retinal or visual pathway disease are difficult to predict as the fundus may be obscured, and assessment of visual potential is challenging. This study assessed the value of visual electrophysiology as a prognostic indicator of visual recovery in diabetic patients with cataract, prior to cataract surgery. Methods: Forty-one diabetic patients (aged 52–80; 74 eyes) and 13 age-matched non-diabetic control patients (21 eyes) were examined prior to cataract surgery. Pre-surgical examinations included best-corrected visual acuity (BCVA), slit-lamp bio-microscopy, ISCEV-standard full-field electroretinography (ffERG), and flash visual evoked potential (flash VEP) testing. Electrophysiological assessments included quantification of the DA and LA ERG, oscillatory potentials (OPs; OP1, OP2, OP3, OP4) and flash VEP P1, P2, and P3 components. Post-operative BCVA was measured in all cases and the diabetic patients grouped according to the severity of visual acuity loss: mild (logMAR ≤ 0.1), moderate (0.1 < logMAR < 0.5), or severe (logMAR ≥ 0.5). A fourth group included those without diabetes. The pre-surgical electrophysiological data was compared between the four groups by analysis of variance. Results: The severity of post-surgical visual acuity loss in the diabetic patients was classified as mild (N=22 eyes), moderate (N=31 eyes), or severe (N=21 eyes). In the group without diabetes, post-surgical visual impairment was classified as mild (N=21 eyes). The pre-operative DA 10.0 ERG a-wave amplitudes, DA 3.0 ERG OP2 amplitudes, and the LA 3.0 a- and b-wave amplitudes showed best significant differences among the four groups. The flash VEP did not show significant difference between groups. Conclusion: Electrophysiological assessment of diabetic patients with cataract can provide a useful measure of retinal function. Full-field ERG components, including the DA 10.0 ERG a-wave, DA 3.0 ERG OP2 component, and the LA 3.0 a- and b-wave amplitudes, are of prognostic value in predicting post-surgical visual acuity, and may inform the surgical management of cataract patients with diabetes. [Figure not available: see fulltext.]

JNK Pathway-Associated Phosphatase/DUSP22 Suppresses CD4+ T-Cell Activation and Th1/Th17-Cell Differentiation and Negatively Correlates with Clinical Activity in Inflammatory Bowel Disease

This study aimed to investigate the role of JNK pathway-associated phosphatase (JKAP) in inflammatory bowel disease (IBD). JKAP expression was analyzed in the intestinal mucosa of 81 IBD patients and 25 healthy controls (HCs) by qPCR and immunoblotting. The correlations of JKAP with clinical activity and inflammatory cytokines were performed. JKAP expression before and after infliximab treatment was also measured. CD4+ T cells were isolated from peripheral blood in active IBD patient and HCs and transduced with lentivirus-encoding JKAP (LV-JKAP), anti-JKAP (LV-anti-JKAP), or empty vector (LV-scramble), and JKAP functions on IBD CD4+ T cells were subsequently investigated. JKAP expression was decreased in inflamed mucosa of active IBD patients and was negatively correlated with disease activity [Crohn’s disease activity index (CDAI), Mayo index, C-reactive protein, and erythrocyte sedimentation rate], interleukin-17, and tumor necrosis factor (TNF)-α levels. Anti-TNF-α treatment up-regulated JKAP expression in CD patients, and baseline JKAP expression was elevated in response patients than in failure patients. Transduction of LV-JKAP into CD4+ T cells inhibited the percentages of CD25+ and CD69+ cells and proliferation. Moreover, inhibition of JKAP promotes Th1/Th17 cell differentiation. Our data indicated that the decreased expression of JKAP in intestinal mucosa contributed to the pathogenesis of IBD, through facilitating CD4+ T-cell activation, proliferation, and Th1/Th17-cell differentiation

Cell cycle arrest mediated by Cd-induced DNA damage in Arabidopsis root tips

Accumulating evidence demonstrates that the aberrant expression of cell cycle regulation and DNA repair genes can result in abnormal cell proliferation and genomic instability in eukaryotic cells under different stresses. Herein, Arabidopsis thaliana (Arabidopsis) seedlings were grown hydroponically on 0.5 × MS media containing cadmium (Cd) at 0–2.5 mg L−1 for 5 d of treatment. Real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis revealed that expression of DNA damage repair and cell cycle regulation genes, including BRCA1, MRE11, WEE1, CDKA;1 and PCNA1, showed an inverted U-shaped dose-response. In contrast, notably reduced expression was observed for G1-to-S transition-related genes, Histone H4, E2Fa and PCNA2; DSB end processing, GR1; G2-to-M transition-related gene, CYCB1;1; and DNA mismatch repair, MSH2, MSH6 and MLH1 genes in root tips exposed to 0.125–2.5 mg/L Cd for 5 d. Flow cytometry (FCM) analysis revealed significant increases of cells with a 2C nuclear content and with a 4C and 8C nuclear content under Cd stresses of 0.125 and 1–2.5 mg L−1, respectively. Our results suggest that 0.125 mg L−1 Cd-induced DNA damage induced the marked G1/S arrest, leading to accelerated growth in root tips, while 1.0–2.5 mg L−1 Cd-induced DNA damage caused a notable G2/M arrest in root tips, leading to reduced growth in root tips. This may be a protective mechanism that prevents cells with damaged DNA from dividing under Cd stress