An animal model of SARS produced by infection of Macaca mulatta with SARS coronavirus

A new SARS animal model was established by inoculating SARS coronavirus (SARS-CoV) into rhesus macaques (Macaca mulatta) through the nasal cavity. Pathological pulmonary changes were successively detected on days 5-60 after virus inoculation. All eight animals showed a transient fever 2-3 days after inoculation. Immunological, molecular biological, and pathological studies support the establishment of this SARS animal model. Firstly, SARS-CoV-specific IgGs were detected in the sera of macaques from 11 to 60 days after inoculation. Secondly, SARS-CoV RNA could be detected in pharyngeal swab samples using nested RT-PCR in all infected animals from 5 days after virus inoculation. Finally, histopathological changes of interstitial pneumonia were found in the lungs during the 60 days after viral inoculation: these changes were less marked at later time points, indicating that an active healing process together with resolution of an acute inflammatory response was taking place in these animals. This animal model should provide insight into the mechanisms of SARS-CoV-related pulmonary disease and greatly facilitate the development of vaccines and therapeutics against SARS.     

Immunohistochemical detection of macrophage-derived foam cells and macrophage colony-stimulating factor in pulmonary atherogenesis of cholesterol-fed rabbits

In order to investigate the role of monocyte/macrophages and their relationship to the expression of macrophage colony-stimulating factor (MCSF) in pulmonary atherosclerosis, lungs were excised from rabbits that had been fed for 60 and 90 days on a diet containing 0.5% cholesterol. In the lungs, fatty streaks and elevated foam cell lesions predominated in the large or medium-sized elastic pulmonary arteries, while massive accumulation of foam cells in the intima of muscular arteries produced marked luminal narrowing and nearly complete occlusion. In these lesions, most of the foam cells were reactive with RbM2, a monoclonal antibody (mAb) against rabbit macrophages, while smooth muscle cell-derived foam cells were detected by mAb against smooth muscle actin in the deeper area of elevated foam cell lesions of elastic arteries. Ultrastructural observation confirmed the presence of monocytes in the intima, their differentiation into macrophages, and their transformation into foam cells in the atherosclerotic lesions. lmmunohistochemical expression of MCSF was demonstrated in the endothelial cells, smooth muscle cells and foam cells. A minor macrophagederived foam cell population was demonstrated to possess a prolif-erative capacity. These data suggest that MCSF is involved in the differentiation of monocytes into macrophages, their transformation into foam cells, and their proliferation during pulmonary atherogenesis.     

Protection by a taurine supplemented diet from lead-induced deficits of long-term potentiation/depotentiation in dentate gyrus of rats in vivo

Previous studies have demonstrated that synaptic plasticity, which includes long-term potentiation (LTP) and depotentiation (DP) in hippocampus, is important for learning and memory. The purpose of this study is to evaluate the effect of taurine via drinking water on the lead-induced impairments of LTP and DP in rat dentate gyrus (DG) in vivo. The experiments were carried out in four groups of rats (control, lead-exposed, control and lead-exposed with a taurine-supplement diet, respectively). The input-output (I/O) function, excitatory postsynaptic potential (EPSP) and population spike (PS) amplitude were measured in the DG area of adult rats (60-90 days) in response to stimulation applied to the lateral perforant path. The results show that: 1. chronic lead exposure impaired LTP/DP measured on both EPSP slope and PS amplitude in DG area of the hippocampus; 2. in control rats, taurine had no effect on LTP/DP; 3. the amplitudes of LTP/DP of lead-exposed group were significantly increased by applying taurine. These results suggest that dietary taurine supplement could protect rats from the lead-induced impairments of synaptic plasticity and might be a preventive medicine to cure the cognitive deficits induced by lead.     

Effect of age on left ventricular systolic function in humans: a study of systolic isovolumic acceleration rate

The aim of this study was to examine the effect of age on left ventricular (LV) systolic function in normal healthy adults. Eighty consecutive subjects without cardiovascular disease underwent standard and tissue Doppler (TD) echocardiographic imaging. LV systolic function was assessed by load-dependent indices as ejection fraction (EF) and myocardial systolic velocities by TD as well as by the load-independent index, systolic isovolumic acceleration rate (IVA). None of the echocardiographic measurements of systolic function declined with age (mean IVA for the group, 286 +/- 123 cm s(-2); IVA vs. age, r = 0.21, P = 0.1). Likewise, LV end diastolic dimension, wall thickness, mass and left atrial maximum volume were not significantly related to age. On the other hand, as previously reported, echocardiographic indices of diastolic function showed a significant decline with age (P < 0.05). When the group was stratified by gender, isovolumic velocity and acceleration were higher in men than women, but the differences were not significant (P = 0.12 and 0.37, respectively). No significant relation was observed between age and measurements of LV systolic function by regression analysis in women (P > 0.1). However, in men, a positive correlation was noted between average IVA and age (r = 0.63, P = 0.007). In conclusion, age is not associated with a change in cardiac structure and LV systolic function, but is accompanied by a decline in echocardiographic indices of diastolic function. With respect to gender, age does not appear to influence LV systolic function in women, but is associated with an increase in IVA and septal systolic ejection in men.     

金丝桃抗脂质过氧化作用研究

为研究金丝桃的抗脂质过氧化作用,用分光光度法测定了金丝桃提取物对大鼠离体组织匀浆的脂质过氧化及由Fe2+-H2O2体系、Fe2+-VitC体系和Fe2+-Cys体系诱导的脂质过氧化的抑制作用。发现金丝桃提取物对大鼠肝、心、肾组织匀浆脂质过氧化和线粒体诱导的脂质过氧化具有抑制作用并表现出剂量依赖关系,而且提取物F003的抑制作用要强于F004、F005的抑制作用。认为金丝桃具有较好的抗脂质过氧化作用,具有进一步研究与开发价值。     

Probucol improves erectile function by regulating endoplasmic reticulum stress in rats with streptozotocin-induced diabetes

This study was to explore the effect and mechanism of Probucol on STZ-induced erectile dysfunction in diabetic rats. Thirty SD male rats aged 12 weeks were given intraperitoneal injection of STZ after fasting for 12 hr. Diabetic rats were haphazardly partitioned under two assemblies and administered 0 or 500 mg/kg probucol by oral gavage to 12 weeks. Control group was intraperitoneally injected with physiological saline, and saline was administered by oral gavage daily. Intracorporeal pressure was used to evaluate erectile function. Levels of proteins were detected using immunohistochemistry and Western blotting. α-SMA and vWF were detected using immunofluorescence staining. After treatment, erectile function in probucol group was significantly improved. Endoplasmic reticulum stress-related proteins were expressed higher in DM group than in sham group, while expression of these proteins decreased significantly in probucol group. However, α-SMA and vWF were expressed at lower levels in DM group than in sham group, and probucol treatment reversed this phenomenon. Finally, Bax and Caspase3 were expressed at higher levels and Bcl-2 was expressed at lower levels in DM group, while the opposite result was obtained in probucol group. In conclusions, probucol improves erectile function by reducing endothelial dysfunction and inhibiting PERK/ATF4/CHOP pathway in STZ-induced diabetic rats.     

瞬时感受器电位离子通道香草素受体4在睾丸缺血再灌注损伤中对GC-1细胞增殖及凋亡的作用及其机制

目的探讨瞬时感受器电位离子通道香草素受体4(TRPV4)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞增殖和凋亡的作用及其机制。方法建立睾丸GC-1细胞缺氧复氧模型,采用蛋白质印迹法(Western blot)检测不同复氧损伤时间点TRPV4的表达变化;分别采用噻唑蓝(MTT)实验、流式细胞术检测转染TRPV4对GC-1细胞增殖和凋亡的影响;采用Western blot法检测睾丸组织中转染TRPV4对GC-1细胞中半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和细胞色素C(Cyt-C)表达的影响,组间比较采用t检验,多组间比较采用单因素方差分析。结果对照组及缺氧复氧组(0、6、12、24、48和72 h)TRPV4的蛋白表达水平分别为0.19±0.02、0.35±0.03、0.42±0.04、0.46±0.04、0.62±0.05、0.54±0.05、0.45±0.04。缺氧复氧组TRPV4表达显著高于未缺氧GC-1细胞的对照组,并在复氧24 h达到峰值(F=6.898,P<0.05),差异有统计学意义;缺氧复氧组中细胞增殖水平明显低于对照组(52.32±4.58比100.00±7.63,t=-9.280,P<0.05),差异有统计学意义,细胞凋亡水平高于对照组(15.60±1.72比4.08±0.87,t=10.352,P<0.05),差异有统计学意义;过表达TRPV4组中细胞增殖水平低于其对照组(23.65±3.98比51.35±4.67,t=-7.820,P<0.05),差异有统计学意义,细胞凋亡水平高于其对照组(26.93±2.15比14.62±1.68),t=7.814,P<0.05),差异有统计学意义;沉默TRPV4组中细胞增殖水平高于其对照组(72.49±6.21比53.18±5.14,t=4.150,P<0.05),差异有统计学意义,细胞凋亡水平低于其对照组(9.71±1.25比15.07±1.64,t=-4.502,P<0.05),差异有统计学意义。缺氧复氧组中Caspase-3和Cyt-C表达水平高于对照组(0.70±0.06比0.20±0.02,t=13.693,P<0.05;0.74±0.07比0.26±0.03,t=10.917,P<0.05),差异有统计学意义;过表达TRPV4组中Caspase-3和Cyt-C表达水平高于其对照组(1.25±0.11比0.69±0.07,t=7.439,P<0.05;1.38±0.14比0.72±0.07,t=7.303,P<0.05),差异有统计学意义;沉默TRPV4组中Caspase-3和Cyt-C表达水平低于其对照组(0.46±0.05比0.68±0.06,t=-4.879,P<0.05;0.45±0.05比0.72±0.06,t=-5.988,P<0.05),差异有统计学意义。结论TRPV4在GC-1细胞中高表达,其可能通过改变GC-1细胞的增殖和凋亡能力,从而影响睾丸IRI的发生发展。     

Targeting NF-κB c-Rel in regulatory T cells to treat corneal transplantation rejection

The relevance of Tregs in the induction of tolerance against corneal allografts has been well established. Although it is well known that the conversion of Tregs into effector-like cells contributes to the loss of corneal immune privilege, the underlying mechanism is still not fully understood. Using heterologous penetrating keratoplasty model, we found that Tregs from corneal allograft rejected mice (inflam-Tregs) exhibit impaired function and characteristics of effector T cells. Further study showed that the expression of NF-κB c-Rel, a key mediator of effector T cell function, was significantly increased in inflam-Tregs. Mechanistic study revealed that elevated NF-κB c-Rel level in inflam-Tregs impaired Treg function through the promotion of inflammatory cytokine production and glycolysis. More importantly, we demonstrated that targeting NF-κB c-Rel was able to improve the immune suppressive function of inflam-Tregs in vitro and enhance the potential of them to suppress corneal transplantation rejection. Therefore, our current study identified NF-κB c-Rel as a key mediator of the conversion of Tregs into effector-like cells when under inflammatory environment.     

Predictive model containing PI-RADS v2 score for postoperative seminal vesicle invasion among prostate cancer patients

BackgroundSeminal vesicle invasion (SVI) is considered to be one of most adverse prognostic findings in prostate cancer, affecting the biochemical progression-free survival and disease-specific survival. Multiparametric magnetic resonance imaging (mpMRI) has shown excellent specificity in diagnosis of SVI, but with poor sensitivity. The aim of this study is to create a model that includes the Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) score to predict postoperative SVI in patients without SVI on preoperative mpMRI.MethodsA total of 262 prostate cancer patients without SVI on preoperative mpMRI who underwent radical prostatectomy (RP) at our institution from January 2012 to July 2019 were enrolled retrospectively. The prostate-specific antigen levels in all patients were <10 ng/mL. Univariate and multivariate logistic regression analyses were used to assess factors associated with SVI, including the PI-RADS v2 score. A regression coefficient-based model was built for predicting SVI. The receiver operating characteristic curve was used to assess the performance of the model.ResultsSVI was reported on the RP specimens in 30 patients (11.5%). The univariate and multivariate analyses revealed that biopsy Gleason grade group (GGG) and the PI-RADS v2 score were significant independent predictors of SVI (all P<0.05). The area under the curve of the model was 0.746 (P<0.001). The PI-RADS v2 score <4 and Gleason grade <8 yielded only a 1.8% incidence of SVI with a high negative predictive value of 98.2% (95% CI, 93.0–99.6%).ConclusionsThe PI-RADS v2 score <4 in prostate cancer patients with prostate-specific antigen level <10 ng/mL is associated with a very low risk of SVI. A model based on biopsy Gleason grade and PI-RADS v2 score may help to predict SVI and serve as a tool for the urologists to make surgical plans.