Real time RT-PCR

实时 RT - PCR
  • 文章类型: Randomized Controlled Trial
    Introduction.我们的合生元(副干酪乳杆菌Shirota,短双歧杆菌菌株Yakult,和低聚半乳糖:LBG)有助于减轻接受新辅助化疗(NAC)的食道癌患者的严重不良事件,如发热性中性粒细胞减少(FN)和腹泻。不幸的是,LBG治疗并不能使所有患者受益。假设/差距声明。在化疗期间与不良事件有关的肠道微生物群物种的鉴定可以帮助预测不良事件的发生。影响LBG功效的肠道微生物群的鉴定也可以帮助建立诊断方法来鉴定在开始治疗之前对LBG有反应的患者。瞄准.确定NAC过程中与不良事件有关的肠道微生物群,并影响LBG治疗的疗效。方法论。这项研究是一项亲本随机对照试验的辅助研究,该试验招募了81名食道癌患者,并给予预防性抗生素或LBG联合肠内营养(LBGEN)。该研究包括81名患者中的73名,这些患者在NAC之前和之后都收集了粪便样本。使用16SrRNA基因扩增子测序分析肠道微生物群,并基于NAC相关不良事件的程度进行比较。此外,本研究还分析了确定的细菌计数和不良事件与LBG+EN缓解效果之间的关联.结果。无FN或仅轻度腹泻的患者的厌氧菌和假双歧杆菌的丰度明显高于FN或严重腹泻的患者(P<0.05)。此外,接受LBG+EN的患者的亚组分析表明,NAC之前的粪便A.hadrus计数与发生FN的风险显着相关(OR,0.11;95%CI,0.01-0.60,P=0.019)。NAC后粪便A.hadrus计数与肠道乙酸(P=0.0007)和丁酸(P=0.00005)的浓度呈正相关。结论。厌氧菌和假性芽孢杆菌可能参与改善不良事件,因此可用于事先识别NAC期间受益于LBGEN的患者。这些结果还表明LBG+EN可用于制定预防NAC期间不良事件的措施。
    Introduction. Our synbiotics (Lacticaseibacillus paracasei strain Shirota, Bifidobacterium breve strain Yakult, and galacto-oligosaccharides: LBG) helps mitigate serious adverse events such as febrile neutropenia (FN) and diarrhoea in oesophageal cancer patients receiving neoadjuvant chemotherapy (NAC). Unfortunately, LBG therapy does not benefit all patients.Hypothesis/Gap Statement. Identification of the gut microbiota species involved in adverse events during chemotherapy could help predict the onset of adverse events. Identification of the gut microbiota that influence the efficacy of LBG could also help establish a diagnostic method to identify patients who will respond to LBG before the initiation of therapy.Aim. To identify the gut microbiota involved in adverse events during NAC and that affect the efficacy of LBG therapy.Methodology. This study was ancillary to a parent randomized controlled trial in which 81 oesophageal cancer patients were recruited and administered either prophylactic antibiotics or LBG combined with enteral nutrition (LBG+EN). The study included 73 of 81 patients from whom faecal samples were collected both before and after NAC. The gut microbiota was analysed using 16S rRNA gene amplicon sequencing and compared based on the degree of NAC-associated adverse events. Furthermore, the association between the counts of identified bacteria and adverse events and the mitigation effect of LBG+EN was also analysed.Results. The abundance of Anaerostipes hadrus and Bifidobacterium pseudocatenulatum in patients with no FN or only mild diarrhoea was significantly higher (P<0.05) compared to those with FN or severe diarrhoea. Moreover, subgroup analyses of patients receiving LBG+EN showed that the faecal A. hadrus count before NAC was significantly associated with a risk of developing FN (OR, 0.11; 95 % CI, 0.01-0.60, P=0.019). The faecal A. hadrus count after NAC was positively correlated with intestinal concentrations of acetic acid (P=0.0007) and butyric acid (P=0.00005).Conclusion. Anaerostipes hadrus and B. pseudocatenulatum may be involved in the ameliorating adverse events and can thus be used to identify beforehand patients that would benefit from LBG+EN during NAC. These results also suggest that LBG+EN would be useful in the development of measures to prevent adverse events during NAC.
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  • 文章类型: Journal Article
    Antimicrobial polypeptides (AMPPs) are humoral components of the vertebrates and invertebrates innate immune system. Their potent broad spectrum antimicrobial activities have drawn the attention of the scientific community to their potential use not only as an alternative to antibiotics but also as functional targets for immunostimulants in order to enhance the host immunity. Fish synthesize a great number of these peptides but in European sea bass, an important fish species in the Mediterranean aquaculture, only a few AMPPs have been studied and these surveys have highlighted their functional role as predictive markers of stressful conditions. Many aspects concerning AMPP mode of action in the host during bacterial infections are still unknown. In this work a 72 h time course experiment, performed on juvenile sea bass intraperitoneally (i.p.) injected with a sub-lethal dose of Vibrio anguillarum, was aimed to investigate the mRNA expression of four specific AMPP genes and interleukin-1β (IL-1β) in skin, gills, spleen, and head kidney. AMPP genes were: dicentracin (DIC), histone-like protein 1 (HLP-1), histone-like protein 2 (HLP-2) and hemoglobin-like protein (Hb-LP). The delta-delta C(T) method in real-time RT-PCR allowed to gain more knowledge about temporal dynamics, preferential sites of expression as well as immunological and physiological role of these molecular markers. DIC was significantly up-regulated mainly in head kidney at 1.5-3 h post-infection (p.i.). HLP-1 showed an extended-time overexpression in gills and a significant up-regulation in spleen. HLP-2 was interestingly overexpressed in gills at 24 h p.i., while Hb-LP showed a significant up-regulation in skin for all the 72 h trial as well as lower but always significant values either in gills or in spleen. Different was the response of IL-1β that showed a dramatic up-regulation in spleen and head kidney at 8 h p.i. whilst in gills it displayed a severe inhibition. During this survey the i.p. stimulus surely conditioned the AMPP expression in skin and gills, especially as regards the DIC that as piscidin-related gene has an important defensive role in the mucosal tissues. However, two unconventional AMPP genes such as HLP-2 and Hb-LP, strictly related to the physiological mechanisms of fish, were less affected in terms of expression by the route of infection, being more evident in peripheral loci. These findings might suggest them as potential markers to be analyzed within plans of health survey in fish farms.
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  • 文章类型: Journal Article
    OBJECTIVE: To investigate the relationship between respiratory viral load and lung lesion severity of patients with pandemic H1N1 2009 influenza A pneumonia.
    METHODS: Cross-sectional observation study.
    METHODS: 24 consecutive H1N1 influenza patients with viral pneumonia (13 males, 11 females, mean age: 17.5 years) during their presentation to hospital were retrospectively analysed. Viral load were first measured on average 5.2 days after the onset of symptoms. The initial CT and viral load measurement was carried on the same day in 13 patients. The rest were carried out with a mean interval time of 1.5 days. All patients had viral load follow-up till turned negative. Thirteen patients had radiological follow-up.
    RESULTS: There was no significant correlation between the initial lung lesion severity and viral load (P=0.4). Both viral load and lung lesion severity decreased over time, being highest value at initial presentation. The patients had higher initial viral load or higher initial lung lesion severity tended to be slower in resolving. The lung lesion decreased at a slower rate than viral load.
    CONCLUSIONS: While there was no correlation between the initial viral load and lung lesion severity, these two indices provide valuable information for epidemiological control.
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