胸水细胞FAK和MMP-9表达在良恶性胸腔积液鉴别中的意义

目的：探讨胸水细胞黏着斑激酶（focaladhesionkinaseFAK）和基质金属蛋白酶-9（MMP-9）检测对良、恶性胸腔积液的鉴别诊断价值。方法：用荧光实时定量RT—PCR和免疫组化方法检测39例恶性、31例良性胸水细胞FAK和MMP-9基因mRNA和蛋白表达,并与胸水细胞学诊断结果进行比较。结果：恶性胸水组FAK和MMP-9mRNA表达水平较良性胸水组和对照明显增加,较良性胸水组分别增加71．9％（P〈0．01）和67．8％（P〈0．01）。免疫组化检测结果显示39例恶性胸水组中FAK基因表达阳性30例（76．9％）,MMP-9基因表达阳性33例（84．6％）,而31良性胸水组FAK基因表达阳性8例（25．8％）,MMP-9基因表达阳性5例（16．1％）（P〈0．01）。结论：胸水细胞MMP-9和FAK基因检测可作为细胞学检查的辅助手段,提高恶性胸腔积液的诊断率。     

Bacterial lipopolysaccharide regulates nociceptin expression in sensory neurons

Nociceptin/orphanin FQ (N/OFQ) is an opioid-related peptide that is markedly up-regulated in sensory neurons in vivo following peripheral inflammation and plays a key role in pain physiology. To identify substances that up-regulate N/OFQ expression in sensory neurons, we carried out an in vitro screen using purified adult mouse dorsal root ganglion (DRG) neurons and identified the potent proinflammatory agent bacterial lipopolysaccharide (LPS) as a very effective inducer of N/OFQ. The robust response of these neurons to LPS enabled us to identify the components of a putative neuronal LPS receptor complex. In contrast to the immune system, where the functional LPS receptor complex is composed of CD-14 together with either MD-2 and TLR4 on myeloid cells or the homologous receptors MD-1 and RP105 on mature B cells, DRG neurons express the unusual combination of CD-14, TLR4, and MD-1. Blocking antibodies against TLR4 and MD-1 prevented induction of N/OFQ by LPS, and, in immunoprecipitation experiments, MD-1 coprecipitated with TLR4. Our findings suggest that LPS regulates N/OFN expression in sensory neurons via a novel combination of LPS receptor components and demonstrate for the first time a direct action of a key initiator of innate immune responses on neurons.     

Effects of nociceptin/orphanin FQ on rats with cathartic colon

AIM To demonstrate the change and effect of nociceptin/orphanin FQ in the colon of rats with cathartic colon.METHODS The cathartic colon model was established by feeding rats rhubarb for 3 mo, the changes of colonic electromyography were investigated by both suspension muscle strips test and serosal recordings of colonic myoelectrical activity. Immunohistochemical staining (S-P methods) and image analysis were used to determine the changes of nociceptin/orphanin FQ in the proximal colon and distal colon of rats with cathartic colon.RESULTS Suspension muscle strips test in vitro showed OFQ (10-9-10-6 mol/L) concentration dependently caused an immediate tonic contraction in the isolated colon. But the increase of tension in cathartic colon was less than control groups (P＜0.01). Intravenous administration of OFQ (1μg/kg) caused phasic contractions in the proximal colon, while the amplitude of phasic contractions caused by OFQ in cathartic colon was much lower than that in the control groups (2.58 ± 0.41 vs 4.16± 0.53, t= -2.6, P = 0.012). OFQ was highly expressed in the myenteric plexus of the rat colon but not in the muscle cells. The immunoreactivity of OFQ in the proximal colon in cathartic colon rats decreased significantly compared with the control group (P = 0.001).CONCLUSION Colonic smooth muscle of cathartic colon showed low sensitivity to the stimulation of OFQ, suggesting that it might be caused by the abnormal distribution of OFQ or the abnormalities of receptors, leading to the disorganization of dynamic and incoordinated contractions.     

Transient receptor potential vanilloid type 1 antagonists: a patent review (2011 – 2014)

Importance of the field: The interactions of nociceptin/orphanin FQ (N/OFQ) and the opioid receptor-like receptor 1 (nociceptin opioid peptide – NOP) have been implicated in a variety of systems including cardiovascular, respiratory, immune, and the central and peripheral nervous systems.

Areas covered in this review: To elucidate the endogenous role of the N/OFQ–NOP system through the use of knockout and knockdown animal preparations, though most advances have been made using a host of synthetic agonists and antagonists. This review gives a brief history of the receptor–ligand discovery, the development of these agonists and antagonists within the last 10 years as published, and the therapeutic indications thereof focusing on pain.

What the reader will gain: The use of NOP ligands in pain has been controversial at best; however, there are indications that both agonists and antagonists have a place in the clinical setting for acute and chronic pain. NOP ligands have potential as novel therapeutics, interestingly, when incorporated into a rationally-designed multi-target agent.

Take home message: The discovery of N/OFQ and NOP opened a new option for the treatment of pain with the potential for a decreased side effect profile. Numerous compounds have been designed to target this system, the most promising of which have mixed profiles.     

Orphanin FQ antagonizes the inhibition of Ca2+ currents induced by μ-opioid receptors

Orphanin FQ (OFQ), an endogenous peptide ligand of opioid receptor-like receptors (ORLs), has properties similar to traditional opioids. This peptide inhibits adenylyl cyclase and voltage-gated calcium channels but stimulates inwardly rectifying potassium channels. Among other actions, however, OFQ also has pharmacological functions that are different from, or even opposite to, those of opioids. For example, OFQ antagonizes the behavioral analgesic effects mediated by κ- and μ-opioid receptors. In a previous paper, we reported that OFQ antagonizes inhibition of calcium channels mediated by κ-opioid receptors. We report here that OFQ also antagonizes the inhibition of calcium channels mediated by μ-opioid receptor. Further, single-cell RT-PCR reveals that the antagonistic effect of OFQ is correlated with the presence of ORL1 mRNA in individual cells.     

Pro-nociceptin/orphanin FQ and NOP receptor mRNA levels in the forebrain of food deprived rats

Forebrain injections of nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the NOP opioid receptor, previously referred to as ORL1 or OP4 receptor, stimulate feeding in freely feeding rats, while the NOP receptor antagonist [Nphe(1)]N/OFQ(1-13)NH(2) inhibits food deprivation-induced feeding. To further evaluate whether the N/OFQ-NOP receptor system plays a physiological role in feeding control, the present study evaluated forebrain mRNA levels for the N/OFQ precursor (pro-N/OFQ), as well as for the NOP receptor in food deprived rats. The results obtained show that food deprived rats have lower mRNA levels for the NOP receptor in several forebrain regions; a significant reduction was found in the paraventricular and lateral hypothalamic nuclei and in the central nucleus of the amygdala. Food deprived rats also exhibited lower pro-N/OFQ mRNA levels in the central amygdala. These results suggest that the N/OFQ-NOP receptor system may have a physiological role in feeding control. The observation that food deprivation reduces gene expression of the N/OFQ-NOP receptor system is apparently not consistent with a direct hyperphagic action for N/OFQ. Taking into account that N/OFQ exerts inhibitory actions at cellular level, the present results may be in keeping with the hypothesis that N/OFQ stimulates feeding by inhibiting neurons inhibitory for food intake; under conditions of food deprivation, these neurons may be silent and the N/OFQ-NOP receptor system, which controls them, may also be regulated at a lower level. Consistently, in the present study N/OFQ stimulated food intake in freely feeding rats, but did not further increase feeding in food deprived rats.     

Effect of opioid and cannabinoid receptor antagonism on orphanin FQ-induced hyperphagia in rats

Feeding induced in rats by cerebroventricular (i.c.v.) injection of orphanin FQ was potently and dose-dependently reversed by peripheral injection of either the opioid antagonist naloxone or the cannabinoid CB(1) receptor antagonist SR 141716[N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophelyl)-4-methyl-3-pyrazole-carboxamine]. The combination of these two agents inhibited food intake in a manner suggestive of additivity or supra-additivity.     

孤儿肽在自发性高血压大鼠和家兔心血管系统的分布

目的 研究孤儿肽（ｏｒｐｈａｎｉｎ ＦＱ,ＯＲＱ）在自发性高血压大鼠和家兔心血管系统的分布。方法 采用反转录－聚合酶链反应（ＲＴ－ＰＣＲ０法和免疫组织化学方法。结果 ＲＴ－ＰＣＲ的结果显示,ＯＦＱ的前体ｍＲＮＡ在自发性高血压大鼠的主动脉,肺动脉,肾动静脉均有较高的表达,其表达水平与脑组织表达水平相似,心房组织也有弱的表达,但在心室组织未见ＯＦＱ前体ｍＲＮＡ的表达。     

荧光定量检测HBV DNA与乙肝两对半结果对比分析研究

目的为了对乙肝患者体内HBV复制及传染性有更直接地了解,亦更有利于对临床HBV感染的诊断、治疗方案的选择及疗效判定.方法运用荧光定量PCR(FQ-PCR)和ELISA两种方法同时检测了310份肝炎患者血清,并对结果进行了对比分析.结果乙肝大三阳患者血清HBV DNA检出率为92.5%(74/80),平均拷贝数为4.10×1010/ml;小三阳患者血清HBV DNA检出率为32.35%(22/68),平均拷贝数为1.31×109/ml;HBsAg(+)、HBcAb(+)组血清HBV DNA检出率为45.71%(32/70),平均考贝数为4.08×108ml;HBV-M全阴性组血清HBV DNA检出率仍达6.67%(2/32),平均拷贝数仍达1.54×108/ml.结论HBV-M阴性患者仍有HBV DNA阳性者,因此对临床HBV感染、复制及传染性的判断以及指导治疗,除乙肝二对半标志物ELISA检测外,加做HBV DNA FQ-PCR检测同样具有重要意义.     

胎儿宫内窘迫孕妇静脉血及脐血孤啡肽的变化

目的探讨宫内窘迫脐血孤啡肽(OFQ)的变化及其临床意义。方法采用放射免疫法测定30例正常妊娠妇女(对照组)及23例胎儿宫内窘迫孕妇(缺氧缺血组)静脉血及脐血OFQ的含量,同时行脐动脉血血气分析。结果1.缺氧缺血组脐血OFQ(23.46±6.72)ng/L,对照组为(13.29±4.14)ng/L,两组相比有显著差异(P<0.05)。2.脐动脉血血气分析,pH 为(7.00±0.15),p(O2)为(1.70±0.42)kPa,p(CO2)为(8.80±0.69)kPa;OFQ与脐血pH、p(O2)呈显著负相关(r= -0.418,-0.437 P均<0.05),与p(CO2)呈显著正相关(r=0.442 P<0.05)。3.缺氧缺血组及对照组母血浆OFQ含量分别为(9.23±3.01)ng/L及(8.57±2.83)ng/L,两组相比无显著差异(P>0.05).4.脐血OFQ含量与母血OFQ含量无显著相关(r=0.287 P>0.05)。结论OFQ与缺氧缺血导致的胎儿窘迫及新生儿窒息的发生发展密切相关。