LDH enzyme activity in human saliva: The effect of exposure to cigarette smoke and its different components

ObjectiveAldehydes and reactive nitrogen species (RNS) are important chemically active agents in cigarette smoke (CS). Salivary lactate dehydrogenase (LDH) originates predominantly from oral epithelium and was identified as an oral state marker. Its activity in saliva decreases after CS exposure. The aims of the current study were to identify the specific damaging agents in CS responsible for this activity reduction and to understand the mechanisms participating in CS oxidative damage to the salivary enzymes.MethodsPurified and salivary LDH samples were exposed to different levels of CS, pure acrolein, acetaldehyde, peroxynitrite and RNS donors. Each response of the isolated agent to the exposure was examined by a spectrophotometric enzyme activity assay and a Western blot.ResultsCS exposure caused a 34% reduction in LDH activity. Isolated treatment with unsaturated-aldehydes (acrolein, 10 μmol) caused a 61% reduction, while saturated-aldehydes (acetaldehyde, 200 μmol), peroxynitrite (200 μM) and RNS donor (SIN-1, 2 mM) caused no substantial effect. All five LDH isoenzymes reacted similarly. The carbonyl immunoblotting assay revealed a fourfold increase in carbonyl content when treated with CS and a sevenfold increase when treated with acrolein.Conclusionα,β-Unsaturated-aldehydes were identified as the main CS ingredient responsible for salivary LDH activity diminution. The effect of saturated-aldehydes and RNS donors was negligible. Unsaturated-aldehydes are capable of introducing carbonyl group into proteins, causing their dysfunction. This provides a molecular explanation for a decrease in LDH enzymatic activity in saliva.     

Effects of electroacupuncture at lower he-sea points on interleukin-1β and high mobility group box 1 in model rats with ulcerative colitis

ObjectiveTo comparatively observe the effect of electroacupuncture at digestive system-related lower he-sea points on the expressions of serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) of colon tissues and high mobility group box 1 protein (HMGB 1) of ulcerative colitis (UC) model rats, and to explore whether there is relative specificity of electroacupuncture at Shàngjùxū (

ST 37), one of lower he-sea points of large intestine, in treatment of bowel diseases.MethodA total of 60 SD rats were randomly divided into control group, model group, ST 37 group, Zúsānlľ (

ST 36) group, Xiàjùxū (

ST 39) group and Yánglíngquán (

GB 34) group. There were ten rats in each group; five were males, and five were females. UC models were established by clysis with 2, 4, 6-trinitrobenzene sulfonic acid/alcohol solution. After modeling, treatment was conducted for ten days, specimens were collected, colonic ulcers and inflammation were inspected visually and scored. The content of serum IL-1β and the expressions of TNF-α and HMGB 1 in colon were detected through ELISA.Results

Compared with control group, the scores of colonic ulcers and inflammation, the content of serum IL-1β and the expressions of TNF-α (except ST 37 group) and HMGB 1 were all higher (P<0.05, P<0.01);

compared with model group, the scores of colonic ulcers in ST 36 group and ST 37 group were lower obviously (P<0.05, P<0.01); the expressions of IL-1β, TNF-α and HMGB 1 in the four treatment groups were lower obviously (P<0.01);

compared with ST 37 group, the expressions of IL-1β, TNF-α and HMGB 1 in other three treatment groups were higher obviously (P<0.05, P<0.01); and the scores of colonic ulcers in ST 39 group and GB 34 group were higher obviously (P<0.05).Conclusion

The score of colonic ulcers can be reduced through electroacupuncture at ST 37, ST 36, ST 39 and GB 34, which can also reduce the content of serum IL-1β and the expressions of TNF-α and HMGB 1, and effectively inhibit inflammatory response of colon caused by UC;

the effect trend of the four acupoints in treatment of UC is: ST 37>ST 36>ST 39>GB 34, and electroacupuncture at ST 37 has the best effect with relative specificity.     

基于药物体系质量评价模式的石菖蒲质量表征关联分析研究

目的基于药物体系的发现,并运用药物体系质量评价模式全面表征石菖蒲质量并关联分析,评价石菖蒲质量。方法采用高效液相色谱法同时测定石菖蒲中原儿茶酸、香草酸、2,4,5-三甲氧基苯甲酸、β-细辛醚、α-细辛醚等5种有效指标性成分的含量。结果基于药物体系质量评价模式对石菖蒲进行有效指标性成分含量及其相对比值的质量表征,并关联分析,使用非关联系数δ、非关联度、关联度等相关概念指标反映与基准饮片样品15质量的关联性。结果表明,样品1、6、9、12、14中有效指标性成分含量总体高于样品15,样品7中有效指标性成分含量与样品15接近。以样品15为基准计算各样品与样品15的非关联系数δ样品7、1、10与样品15质量关联度较高。综合质量表征及关联分析结果,样品1、7饮片质量优良度居前,其次为样品12、14、9、6。结论基于药物体系质量评价模式对石菖蒲质量进行关联分析研究,以具有确切药效的饮片为基准,结合关联度分析,可综合精准地评价出石菖蒲质量优次。     

Pathogenic molecular mechanisms in an animal model of fulminant hepatic failure: rabbit hemorrhagic viral disease

In this study we sought to determine whether molecular mechanisms involved in the pathogenesis of fulminant hepatic failure are present in rabbits experimentally infected with rabbit hemorrhagic disease virus (RHDV). The activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase, as well as bilirubin concentration, were found to be significantly increased 36 hours after infection. Infected animals also demonstrated significant decreases in factor VII activity, in the Fischer index, and in the deterioration of prothrombin time. The concentration of reduced glutathione was significantly decreased 36 hours after infection, and we noted a marked increase in the ratio of oxidized to reduced glutathione. Infected animals showed progressive decreases in liver activity of the antioxidant enzyme superoxide dismutase. Expression of hepatocyte growth factor and c-met was found to be progressively reduced from 24 hours after infection, during which time we detected no modification in messenger RNA (mRNA) levels of transforming growth factor (TGF)-alpha. TFG-beta 1 was overexpressed 24 and 36 hours after infection, and 36 hours after infection we detected a significant increase in TNF-alpha mRNA levels. Experimental RHDV infection also induced marked activation of nuclear factor-kappaB and a significant increase in inducible nitric oxide synthase mRNA levels from 24 hours after infection. Data obtained from this animal model support its usefulness in the investigation of potential novel therapeutical modalities aimed at neutralizing reactive oxygen species and hepatocyte growth inhibitors or enhancing hepatocyte responsiveness to mitogens.     

α-氰基丙烯酸酯类医用黏合剂的研究进展

α-氰基丙烯酸酯类化合物可作为快速黏合剂,广泛应用于医疗领域,用于止血、伤口黏合、骨骼黏接、医疗栓塞、无痛绝育等。α-氰基丙烯酸酯化合物的合成路线各有优缺点,应结合结构特点进行选择。通过结构修饰可期望改善性能,扩大应用。本文综述了α-氰基丙烯酸酯类化合物的合成方法,结构修饰以及在医疗领域的研究进展,并展望在军事医学方面的应用前景。     

Patient-specific decision modeling to guide the use of drotrecogin α (activated) in patients with severe sepsis

Purpose

The expected benefit of treating severe sepsis with drotrecogin α (activated) for an individual patient may depend upon several clinical factors including disease severity. Our objective was to create a decision support tool incorporating patient-specific inputs to estimate the balance between treatment risks and benefits for individual patients with severe sepsis.

Materials and Methods

Logistic regression models were developed to calculate patient-specific mortality risk with and without treatment, which were then used as inputs into a 75-state Markov model. Patient-specific inputs included patient age, sex, and 12 readily available clinical characteristics.

Results

The expected benefit from drotrecogin α (activated) treatment was most dependent upon the underlying disease severity. For example, for a 56-year-old white man with severe sepsis and a 28-day mortality risk of 29%, the model predicted a treatment-related gain of 1.2 quality-adjusted life years (17.3 vs 16.1). Probabilistic sensitivity analyses demonstrated that this patient would benefit from therapy 85% of the time.

Conclusions

A customizable decision model using patient-specific inputs can be used to inform the treatment decision when considering the use of drotrecogin α (activated) therapy by weighing the risks vs the benefits of therapy in the treatment of severe sepsis.     

低氧诱导因子-1α和血管内皮生长因子在玉树地震救援人员急性高原反应低氧性肺动脉高压发生中的作用

目的 观察低氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)在急性高原反应(AHAR)低氧性肺动脉高压(HPH)发生中的作用及返回低海拔后的变化。方法 选择2010年4月14日青海玉树大地震后由低海拔(1 500m)快速进入高海拔地区(3 700m)并从事重体力劳动的18～35岁男性官兵96名,根据AHAR症状评分分为无AHAR组(25名)、轻中度AHAR组(47名)和重度AHAR组(24名);在高海拔地区停留50 d后下撤前及返回低海拔地区后12h、15d分别测定平均肺动脉压(mPAP)和血清HIF-1α、VEGF水平。同时选择低海拔地区50名健康官兵作为对照。结果 高海拔无AHAR组mPAP(mm Hg,1 mm Hg=0.133 kPa)和血清HIF-1α(pg/L)、VEGF(ng/L)水平（分别为24.23±1.56、68.80±7.52、82.56±6.32）明显高于低海拔对照组(18.50±1.30、50.95±3.33、65.78±4.03);且随AHAR加重,各指标进一步升高,高海拔轻中度AHAR组分别为28.42±1.32、88.10±9.20、104.82±10.36,重度AHAR组分别为34.70±2.94、117,93±13.46、136.77±12.03,组间两两比较差异均有统计学意义(均P＜0.01)。高海拔AHAR总计分与mPAP、血清HIF-1α、VEGF水平均呈显著正相关(r值分别为0.672、0.737、0.634,均P＜0．01);mPAP与血清HIF- 1α、VEGF水平呈显著正相关(r值分别为0.706、0.638,均P＜0.01)。与低海拔对照组比较,96名官兵进入高海拔地区50 d时mPAP (mm Hg)和血清HIF- 1α (pg/I)、VEGF (ng/L)水平（分别为29.08±4.22、91.16±20.58、107.11±10.32）显著升高(均P＜0.01),返回低海拔地区12 h(分别为23.05±3.18、70.99±8.22、78.65±6.47)、15 d（分别为18.96±1.75、52.31±4.92、63.08±4.55）时各指标显著下降,且15d明显低于12 d(均P＜0．01),15d时各指标与低海拔对照组比较差异均无统计学意义(均P＞0.05)。结论 人体在高海拔低氧地区并从事重体力劳动时,AHAR越重,mPAP和血清HIF-1α、VEGF水平就越高,说明HPH与HIF-1α、VEGF水平升高有密切关系;返回低海拔地区后12h上述指标有显著改善,15d可恢复到正常水平。     

Tumor necrosis factor-α induces sensitization of meningeal nociceptors mediated via local COX and p38 MAP kinase actions

The proinflammatory cytokine TNF-α has been shown to promote activation and sensitization of primary afferent nociceptors. The downstream signaling processes that play a role in promoting this neuronal response remain however controversial. Increased TNF-α plasma levels during migraine attacks suggest that local interaction between this cytokine and intracranial meningeal nociceptors plays a role in promoting the headache. Here, using in vivo single unit recording in the trigeminal ganglia of anesthetized rats, we show that meningeal TNF-α action promotes a delayed mechanical sensitization of meningeal nociceptors. Using immunohistochemistry, we provide evidence for non-neuronal localization of the TNF receptors TNFR1 to dural endothelial vascular cells and TNFR2 to dural resident macrophages as well as to some CGRP-expressing dural nerve fibers. We also demonstrate that meningeal vascular TNFR1 is co-localized with COX-1 while the perivascular TNFR2 is co-expressed with COX-2. We further report here for the first time that TNF-α evoked sensitization of meningeal nociceptors is dependent upon local action of cyclooxygenase (COX). Finally, we show that local application of TNF-α to the meninges evokes activation of the p38 MAP kinase in dural blood vessels that also express TNFR1 and that pharmacological blockade of p38 activation inhibits TNF-α evoked sensitization of meningeal nociceptors. Our study suggests that meningeal action of TNF-α could play an important role in the genesis of intracranial throbbing headaches such as migraine through a mechanism that involves at least part activation of non-neuronal TNFR1 and TNFR2 and downstream activation of meningeal non-neuronal COX and the p38 MAP kinase.     

Short term exendin-4 treatment reduces markers of metabolic disorders in female offspring of obese rat dams

ObjectivesMaternal obesity imposes significant health risks in the offspring including diabetes and dyslipidemia. We previously showed that the hypoglycaemic agent exendin-4 (Ex-4) administered from weaning can reverse the maternal impact of ‘transmitted disorders’ in such offspring. However daily injection for six-weeks was required and the beneficial effect may lapse upon drug withdrawal. This study aimed to investigate whether short term Ex-4 treatment during suckling period in a rodent model can reverse transmitted metabolic disorders due to maternal obesity.MethodsMaternal obesity was induced in female Sprague Dawley rats by high-fat diet feeding for 6 weeks, throughout gestation and lactation. Female offspring were treated with Ex-4 (5 μg/kg/day) between postnatal day (P) 4 and 14. Female offspring were harvested at weaning (P20). Lipid and glucose metabolic markers were measured in the liver and fat. Appetite regulators were measured in the plasma and hypothalamus.ResultsMaternal obesity significantly increased body weight, fat mass, and liver weight in the offspring. There was an associated inhibition of peroxisomal proliferator activated receptor gamma coactivator 1α (PGC1α), increased fatty acid synthase (FASN) expression in the liver, and reduced adipocyte triglyceride lipase (ATGL) expression. It also increased the plasma gut hormone ghrelin and reduced glucagon-like peptide-1. Ex-4 treatment partially reversed the maternal impact on adiposity and impaired lipid metabolism in the offspring, with increased liver PGC1α and inhibition of FASN mRNA expression. Ex-4 treatment also increased the expression of a novel fat depletion gene a2-zinc-glycoprotein 1 in the fat tissue.ConclusionShort term Ex-4 treatment during the suckling period significantly improved the metabolic profile in the offspring from the obese mothers at weaning. Long-term studies are needed to follow such offspring to adulthood to examine the sustained effects of Ex-4 in preventing the development of metabolic disease.     

天然罗汉松二萜体外抗登革病毒活性研究

登革病毒(dengue virus,DENV)是造成热带和亚热带地区登革热和登革出血热季节性大爆发的蚊媒病原体,可导致严重威胁生命的疾病,因此亟需研发DENV疫苗和药物.本研究发现从石栗枝叶中分离的罗汉松型二萜(3α,5β,10α)-13-methoxypodocarpa-8,11,13-triene-3,12-dio...