Cells infected with herpes simplex virus 1 export to uninfected cells exosomes containing STING,viral mRNAs,and microRNAs

STING (stimulator of IFN genes) activates the IFN-dependent innate immune response to infection on sensing the presence of DNA in cytosol. The quantity of STING accumulating in cultured cells varies; it is relatively high in some cell lines [e.g., HEp-2, human embryonic lung fibroblasts (HEL), and HeLa] and low in others (e.g., Vero cells). In a preceding publication we reported that STING was stable in four cell lines infected with herpes simplex virus 1 and that it was actively stabilized in at least two cell lines derived from human cancers. In this report we show that STING is exported from HEp-2 cells to Vero cells along with virions, viral mRNAs, microRNAs, and the exosome marker protein CD9. The virions and exosomes copurified. The quantity of STING and CD9 exported from one cell line to another was inoculum-size–dependent and reflected the levels of STING and CD9 accumulating in the cells in which the virus inoculum was made. The export of STING, an innate immune sensor, and of viral mRNAs whose major role may be in silencing viral genes in latently infected neurons, suggests that the virus has evolved mechanisms that curtail rather than foster the spread of infection under certain conditions.The stimulator of IFN genes (STING) is a sensor of cytoplasmic DNA and activates immune responses to intracellular pathogens (1–3). Knockout of STING exacerbates the pathogenicity of herpes simplex virus (HSV-1) and of other pathogens in mice (2, 3). Two observations reported earlier add to the role of STING in HSV-1 infected cells. Specifically, (i) STING was readily detectable and stable in four different cell lines infected with wild-type virus (4). The stability of STING in cells infected with an HSV-1 mutant lacking the gene encoding ICP0 (infected cell protein no. 0), a key viral regulatory protein, varied. STING was stable in ΔICP0 mutant virus infected cells derived from normal tissues but was rapidly degraded in infected cells derived from human cancers (4). Implicit in this observation is that ICP0 is required to stabilize STING, although STING could also be stabilized in the absence of ICP0 by a cellular function. (ii) Depletion of STING increased wild-type virus yield 10-fold in cells derived from normal tissues but decreased the yield by the same amount in cells derived from human cancer (4). Thus, at least in cells derived from normal tissues, HSV-1 expresses ICP0 that enables the stabilization of STING even though STING is inimical to virus growth. The results of these studies suggest that HSV-1 recruits STING for a specific function, even though the persistence of STING is not beneficial to virus growth, at least in cells derived from normal tissues (4).Here we report that STING, along with viral RNAs contained in structures that coprecipitate with exosome marker proteins, is exported from the cells in which virus is produced to uninfected cells. The quantities introduced into uninfected cells are dose-dependent and reflect the amounts produced in donor cells.Cells continuously secrete a large number of microvesicles, macromolecular complexes, and small molecules into the extracellular space (5, 6). Of the secreted microvesicles, exosomes are 30–120 nm in diameter, containing nucleic acid and proteins, and are perceived to be carriers of this cargo between diverse locations. They are distinguished in their genesis by being budded into endosomes to form multivesicular bodies (MVBs) in the cytoplasm. The exosomes are released to extracellular fluids by fusion of these multivesicular bodies with the cell surface, resulting in secretion (7–10).Several pathogens use the exosomes to manipulate their microenvironment (11, 12). Viruses, especially small retroviruses such as HIV, use the exosome pathway for egress and immune evasion (13–15). The hepatitis C virus uses exosomes for invasion and spread (16–18). EBV-infected cells release exosomes containing the latent membrane protein 1 to induce T-cell anergy (19–23). In the case of human cytomegalovirus the exosomes carrying viral antigens exacerbate the transplant rejection process (24). It is likely that viruses that establish long-term, latent, or chronic infections modulate exosomes to enhance their persistence (11).Here we report that the exosomes secreted by HSV-1–infected cells deliver to uninfected cells the innate immune sensor STING along with viral RNAs. We speculate that in the long run the strategy serves the virus to fulfill its mission: to spread effectively from person to person.     

股骨颈骨折内固定失败后行全髋关节置换术治疗的效果分析

目的分析探讨股骨颈骨折内固定失败后行全髋关节置换术(total hip arthroplasty,THA)治疗的临床疗效。方法回顾性分析2018年1月至2019年1月南阳市骨科医院关节科收治的股骨颈骨折内固定失败后行THA治疗的36例患者的病历资料,对比THA治疗前后髋关节Harris评分及简明健康状况调查量表(SF-36)评分变化情况,评估患者髋关节功能及生活质量。结果治疗后6个月患者髋关节Harris评分为(66.21±6.42)分,治疗后12个月为(88.45±7.19)分,均明显高于治疗前的(43.11±7.25)分(t=14.310、26.640,P均=0.000);治疗后12个月,患者简明健康状况调查量表(SF-36)中的躯体功能、躯体疼痛、生理职能、社会功能、精神健康、情感职能、精力、一般健康状况评分均明显高于治疗前(t=12.790、22.190、7.367、12.760、12.630、6.332、12.160、12.160,P均=0.000)。结论股骨颈骨折内固定失败后予以THA治疗,能够有效提高患者的髋关节功能及生活质量,临床应用价值较高。     

Depression in older patients with advanced colorectal cancer is closely connected with immunosuppressive acidic protein

Colorectal cancer (CRC) is one of the most common tumors. CRC patients are susceptible to suffering from depression. Whether the immune system of CRC patients with depression is impaired or stimulated is controversial. Possible reasons for this conflict are the involvement of confounding factors, such as the age of the patient, the stage of the CRC and the types of treatment in previous studies. To demonstrate clearly the relationship between depression and the immune system in the context of CRC, the present study included only older patients with advanced CRC who received only chemotherapy, and the study adopted immunosuppressive acidic protein (IAP) as an immune parameter for the first time. A total of 56 older patients with advanced CRC completed the Zung Self-Rating Depression Scale (SDS) and were divided into two groups according to SDS scores. The patients exhibiting depression were treated with fluoxetine until their symptoms remitted. The serum levels of IAP and the percentages of CD3-positive (CD3+), CD4+, CD8+ T lymphocytes and CD56+ natural killer (NK) cells and Neutrophil-lymphocyte ratio (NLR) were calculated at the time of enrollment and once the symptoms remitted. Correlation analyses revealed that the SDS score was positively associated with serum IAP levels but negatively associated with CD3 and CD4 levels. Among the depressed and non-depressed patients, serum IAP levels and the percentages of CD3 and CD4 cells were dramatically different. After the depression symptoms were treated, the IAP levels dramatically decreased, while the levels of CD3, CD4, CD8 and CD56 were unchanged. All of above suggested that IAP was closely correlated with depression and might be a relatively objective parameter for predicting depression.     

LipoxinA4 attenuates myocardial ischemia reperfusion injury via a mechanism related to downregulation of GRP-78 and caspase-12 in rats

This study aims to determine the effect of Lipoxin (LX)A₄ on myocardial ischemia reperfusion injury (MIRI) in rats and the related molecular mechanisms. Male SD rats were divided into six groups. The sham operation groups (groups C1, C2) were injected with 2 ml/kg normal saline before and after coronary artery threading, respectively. The MIRI group (groups I/R1, I/R2) were injected with normal saline before and after MIRI, respectively. The LXA₄ groups (groups LX1, LX2) were injected with LXA₄ before and after MIRI treatment, respectively. The hematoxylin–eosin staining and ultrastructural changes of cardiac muscle were observed. The serum levels of interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor (TNF) α and cardiac troponin I (cTnI) were measured before open-chest operation and at the end of the experiment. The mRNA and protein levels of GRP-78 and caspase-12 were determined in each group. The myocardial cell apoptosis, myeloperoxidase (MPO), superoxide dismutase (SOD), and malondialdehyde (MDA) contents were detected. The mRNA and protein levels of GRP-78 and caspase-12, the apoptosis, the serum IL-1β, IL-6, IL-10, TNF-α, and cTnI concentrations, MPO, SOD, MDA contents were significantly increased in groups I/R1, I/R2, LX1, and LX2 compared with those in groups C1 and C2 (P < 0.05). The mRNA and protein expression levels of GRP-78 and caspase-12 in groups LX1 and LX2 were lower than those in groups I/R1 and I/R2. Compared with group I/R1 and I/R2, the myocardial neutrophil infiltration and ultrastructure damage were significantly less in groups LX1 and LX2. GRP-78 and IL-10 are expressed both extracellularly and intracellularly, but are mainly expressed in the cytoplasms. In the absence of MIRI, LXA₄ has no detectable effect on GRP-78 and caspase-12 expression. Before and after MIRI, application of LXA₄ significantly inhibits neutrophil activation, and attenuates myocardial inflammatory injury and oxidative stress. LXA₄ downregulates the mRNA and protein expression of GRP-78 and caspase-12. LXA₄ could play a role in myocardial protection via a mechanism related to downregulation of GRP-78 and caspase-12, and inhibition of apoptosis.     

Vasodilatory Effect of a Novel Rho-Kinase Inhibitor,DL0805-2, on the rat Mesenteric Artery and its Potential Mechanisms

Purpose

In the present study, we investigated the vasodilatory effect of a novel scaffold Rho-kinase inhibitor, DL0805-2, on isolated rat arterial rings including mesenteric, ventral tail, and renal arteries. We also examined the potential mechanisms of its vasodilatory action using mesenteric artery rings.

Methods

A DMT multiwire myograph system was used to test the tension of isolated small arteries. Several drugs were employed to verify the underlying mechanisms.

Results

DL0805-2 (10^?7–10^?4 M) inhibited KCl (60 mM)-induced vasoconstriction in three types of small artery rings (pEC₅₀: 5.84?±?0.03, 5.39?±?0.03, and 5.67?±?0.02 for mesenteric, renal, and ventral tail artery rings, respectively). Pre-incubation with DL0805-2 (1, 3, or 10 μM) attenuated KCl (10–60 mM) and angiotensin II (AngII; 10^?6 M)-induced vasoconstriction in mesenteric artery rings. The relaxant effect on the rat mesenteric artery was partially endothelium-dependent (pEC₅₀: 6.02?±?0.05 for endothelium-intact and 5.72?±?0.06 for endothelium-denuded). The influx and release of Ca²⁺ were inhibited by DL0805-2. In addition, the increased phosphorylation levels of myosin light chain (MLC) and myosin-binding subunit of myosin phosphatase (MYPT1) induced by AngII were blocked by DL0805-2. However, DL0805-2 had little effect on K⁺ channels.

Conclusions

The present results demonstrate that DL0805-2 has a vasorelaxant effect on isolated rat small arteries and may exert its action through the endothelium, Ca²⁺ channels, and the Rho/ROCK pathway.     

Hemoglobin A1c as a marker for identifying diabetes and cardiovascular risk factors: the China Health and Nutrition Survey 2009

Hemoglobin A_1c (HbA_1c) has been recommended as an optional method for diagnosing diabetes. The impact of HbA_1c on the diagnosis of diabetes has not been evaluated in China, a country with the greatest number of people with diabetes in the world. Hence, we aim to examine how well HbA_1c performs as compared with fasting plasma glucose (FPG) for diagnosing diabetes in Chinese population. We conducted a cross-sectional analysis of 7,641 Chinese men and women aged ≥18 years using data from the China Health and Nutrition Survey 2009 in which FPG and standardized HbA_1c were measured. HbA_1c was measured with high-performance liquid chromatography system. Diabetes is defined as having FPG ≥7 mmol/l or HbA_1c ≥6.5 %. Overall, 5.0 and 5.8 % had undiagnosed diabetes by FPG ≥7 mmol/l and HbA_1c ≥6.5 %, respectively. Overlap between HbA_1c- and FPG-based diagnosis of diabetes was limited (n = 214, 34.9 %). Similar trends were noted in both genders, all age groups, urban/rural settings, regions, body mass index (BMI) categories, waist circumference (WC) groups, and blood pressure status. Solely HbA_1c-defined individuals exhibited higher levels of BMI, WC, total cholesterol, and hypersensitive C-reactive protein and lower levels of homeostasis model assessment of insulin resistance. We note limited overlap between FPG- and HbA_1c-based diagnosis of diabetes. The limited overlap between FPG- and HbA_1c-based diagnosis of diabetes persisted in each evaluated subgroup. HbA_1c criterion for the diagnosis of diabetes identifies individuals with a worse cardiovascular risk profile compared with FPG.     

Magnetic resonance imaging and risk factors for progression of lacunar infarct lesions in Chinese patients

Neuroradiology - The proportion of acute symptomatic lacunar infarction lesions that undergo cavitation and the factors influencing cavity formation are yet unclear, particularly in the Chinese...