The effect of functional gastrointestinal disorders on psychological comorbidity and quality of life in patients with inflammatory bowel disease

Background  Symptoms of functional gastrointestinal disorders (FGIDs) are common in patients with inflammatory bowel disease (IBD). Psychological comorbidities of anxiety and depression are also highly prevalent in IBD.
Aim  To quantify the burden of FGIDs in a hospital-based cohort of patients with IBD and to determine whether there is any inter-relationship between the presence and number of FGIDs and patients' quality of life or psychological status.
Methods  A cross-sectional survey of 61 out-patients was performed. Data on psychological status, quality of life, disease activity and functional symptoms according to Rome III criteria were collected.
Results  Overall, 49 (80%) participants met Rome III criteria for a functional bowel disorder and 52% of participants met criteria for more than one FGID. Participants with no FGID had significantly better physical quality of life than those with more than two FGIDs ( P  = 0.025). However, there was no relationship among the number of FGIDs, mental quality of life, anxiety or depression.
Conclusions  Functional gastrointestinal disorders are highly prevalent in out-patients with IBD. Somewhat unexpectedly, the presence of anxiety and/or depression did not appear to correlate with either the presence or the number of FGIDs.     

Duodenal mastocytosis, eosinophilia and intraepithelial lymphocytosis as possible disease markers in the irritable bowel syndrome and functional dyspepsia

Background  Irritable bowel syndrome (IBS) and functional dyspepsia (FD) are common functional disorders without defined pathology. Mast cells and eosinophils interact with T lymphocytes and may alter enteric nerve and smooth muscle function.
Aim  To examine mast cell, eosinophil and intraepithelial lymphocyte populations in duodenal biopsies of subjects with IBS and FD.
Methods  A random sample of an adult Swedish population ( n  = 1001; mean age 54 years; 51% female) underwent upper endoscopy and biopsy; 51 cases with FD and 41 cases with IBS were compared with 48 randomly selected controls. Eosinophils were identified by light microscopy; mast cells by immunocytochemistry (CD117). Intraepithelial lymphocytes were counted per 100 enterocytes. Cell counts were quantified by counting the number per high power field (HPF) in 5HPFs in the bulb (D1) and second part of duodenum (D2), summed over 5HPFs at each site.
Results  Cases and controls showed similar demographics. Compared to controls, IELs in IBS-constipation were significantly increased ( P  = 0.005). Mast cells were significantly increased in IBS in D2 ( P  < 0.001), while eosinophils were significantly increased in FD in D1 and D2 ( P  < 0.001).
Conclusion  Duodenal mast cell hyperplasia is linked to IBS and eosinophilia to FD, and duodenal biopsy may identify subsets of these disorders.     

人巨细胞病毒感染的核酸检测比较及对肝脏损伤的研究

目的：应用实时荧光定量聚合酶链反应（ FQ-PCR）检测疑似人巨细胞病毒（ HCMV）感染的患儿尿液、血液及对应母亲乳汁中HCMV DNA含量,探讨其在HCMV感染诊断中的价值及对肝脏损伤情况。方法应用FQ-PCR方法检测104例疑似HCMV感染患儿新鲜尿液上皮细胞、血液及其对应的母亲乳汁中的HCMV DNA拷贝数,并同时检测患儿血液中天冬氨酸转氨酶（ AST）、丙氨酸转氨酶（ ALT）、谷氨酰转肽酶（GGT）、总胆红素水平。结果尿液、血液标本阳性检出率分别为52．9％（55/104）,24．0％（25/104）,尿液阳性率明显高于血液阳性率,二者差异有统计学意义（P＜0．01）。乳汁标本阳性检出率为81．7％（85/104）,对应患儿血液和/或尿液阳性率为62．4％（53/85）,二者差异有统计学意义（P＜0．05）。且患儿HCMV拷贝数越高,肝脏损伤的可能性越大。结论婴儿尿液中HCMV DNA检测率明显高于血液的检出率,且拷贝数越高,肝功能损伤的可能性就越大。尿液HCMV DNA检测对于诊断婴儿HCMV感染和治疗监测具有重要的价值。肝功能的检测对判断HCMV感染严重程度有一定的参考价值。     

Highly specific,quantitative polymerase chain reaction probe for the quantification of human cells in cynomolgus monkeys

Quantification of human cells may be performed using quantitative polymerase chain reaction (qPCR). In preclinical studies, the human Alu sequence is widely used as biomarker for human DNA. However, because the Alu gene is shared by primates, its use is limited to non-primate studies. The biodistribution of human cells in primates is also necessary for translational studies. Therefore, we aimed to design a novel, human-specific primer/probe that enables the quantification of human cells in primates and other animal models. A novel primer/probe set was successfully designed based on highly repetitive LINE1 sequences. qPCR efficiency (94.95–99.21%) and linearity of calibration curves (r² = 0.996–0.999) were confirmed in tissue homogenates of cynomolgus monkey. The lower limit of detection was 10 cells per 15-mg tissue sample, a sensitivity that is equivalent to existing Alu primers/probes. The set was also effective in other animal models such as mice, rabbits, pigs, and common marmosets.To our knowledge, this is the first study describing the successful design of a human-specific qPCR primer/probe for human cell quantification in various animals, including non-human primates, using LINE1 sequence. The excellent selectivity, sensitivity, and versatility of the LINE1 primers/probes make it a promising quantification tool in preclinical biodistribution studies.     

No evidence of involvement of Chlamydia pneumoniae in lung cancer by means of quantitative real-time polymerase chain reaction

Chlamydia pneumoniae, an obligate intracellular pathogen, is well-known as etiological agent of acute respiratory infections; the repeated or prolonged exposure to chlamydial antigens may promote the persistence of C. pneumoniae in the respiratory tract leading to chronic diseases, such as chronic obstructive pulmonary disease and asthma. The predilection of C. pneumoniae to cause respiratory tract infections combined with its persistent nature suggest that it might play a role in lung cancer. The aim of our study is to evaluate the involvement of C. pneumoniae in pathogenesis of lung cancer. We therefore investigated the presence of C. pneumoniae DNA in tumor lung tissues by using real-time PCR assay. Simultaneously, tumor and healthy tissues from the same patient with primary carcinoma lung were analyzed. C. pneumoniae DNA was not detected in a single lung tumor tissue by means of an highly sensitive, and specific real-time PCR assay based on FRET hybridization probes. In conclusion, this study does not support the involvement of C. pneumoniae in the pathogenesis of lung cancer, suggesting that further investigations are needed to clarify other potential causative factors for the development of this malignancy.     

荧光定量聚合酶链反应检测白细胞中结构分枝杆菌DNA

目的：检测外周血白细胞中结核分枝杆菌DNA在肺结核诊断中的价值。方法：应用荧光定量聚合酶链反应技术，对95例肺结核患者外周血标本，84例肺结核患者痰标本进行了结核分枝杆菌DNA检测。结果：外周血、痰标本中，结核分枝杆菌DNA阳性率为85％和57％；对44例患者外周血、痰标本配对同时检测总阳性率可达91％。结论：荧光定量聚合酶链反应检测外周血白细胞中结核杆菌DNA用于肺结核的诊断，可提高肺结核诊断的敏感性和准确性，明显优于痰标本，可作为肺结核诊断较可靠指标。     

Review article: gastrointestinal sensory and motor disturbances in inflammatory bowel disease - clinical relevance and pathophysiological mechanisms

Background It is well known that inflammation has a profound impact on the neuromuscular apparatus of the gastrointestinal tract during the inflammatory insult and in periods of remission, at the site of inflammation and at distance from this site. The importance of this interaction is illustrated by the higher prevalence of functional gut disorders in patients with inflammatory bowel disease. Aims To document the epidemiological and clinical significance of functional alterations of gut motility and sensitivity in patients with inflammatory bowel disease and to formulate potential pathophysiological mechanisms. Results and conclusions Functional gut disorders occur frequently in patients with inflammatory bowel disease, both during inflammatory episodes and in periods of remission, and have a major impact on their quality of life. The clinical manifestations of these motility and sensitivity disorders vary and are often difficult to treat, mainly because therapeutic guidelines and specific diagnostic tests to distinguish inflammatory bowel disease from functional gut disorders are lacking. Chronic bowel inflammation results in a complicated interaction between neuroendocrine serotonin‐predominant cells of the mucosa, inflammatory cells (particularly mast cells) in the submucosa, the intrinsic and extrinsic innervation and the muscular apparatus including the interstitial cells of Cajal. The outcome of this interaction is a perturbation of gastrointestinal motor function, both locally and at distance from the site of inflammation and during both acute inflammation and remission.     

Effects of a bicarbonate-alkaline mineral water on digestive motility in experimental models of functional and inflammatory gastrointestinal disorders

This study investigates the effects of Uliveto, a bicarbonate-alkaline mineral water, in experimental models of diarrhea, constipation and colitis. Rats were allowed to drink Uliveto or oligomineral water (control) for 30 days. Diarrhea and constipation were evoked by 16,16-dimethyl-prostaglandin E(2) (dmPGE(2)) or loperamide, respectively. Colitis was induced by 2,4-dinitrobenzenesulfonic acid (DNBS) or acetic acid. Gastric emptying, small-intestinal and colonic transit were evaluated. dmPGE(2)-induced diarrhea reduced gastric emptying and increased small-intestinal and colonic transit. In this setting, Uliveto water enhanced gastric emptying, and this effect was prevented by L-365,260 (gastrin receptor antagonist). Loperamide-induced constipation reduced gastric emptying, small-intestinal and colonic transit, and these effects were prevented by Uliveto water. L-365,260 counteracted the effects of Uliveto on gastric emptying, while alosetron (serotonin 5-HT(3) receptor antagonist) blunted the effect of Uliveto on colonic transit. Gastric emptying, small-intestinal and colonic transit were reduced in DNBS-induced colitis, and Uliveto water enhanced gastric emptying and normalized small-intestinal and colonic transit. Gastric emptying, small-intestinal and colonic transit were also reduced in acetic acid-induced colitis, and Uliveto increased both gastric emptying and small-intestinal transit. In conclusion, Uliveto water exerts beneficial effects on gastrointestinal motility in the presence of bowel motor dysfunctions. The effects of Uliveto water on gastric emptying depend on gastrin-mediated mechanisms, whereas the activation of serotonergic pathways accounts for the modulation of colonic functions.     

The selection and identification of compound housekeeping genes for quantitative real-time polymerase chain reaction analysis in rat fetal kidney

During quantitative real-time polymerase chain reaction (RT-qPCR) data analysis, the selection of optimal housekeeping gene is necessary to ensure the accuracy of results. It is noteworthy that housekeeping genes commonly used in adult studies may not be applicable for fetus. However, the stability analysis of housekeeping gene in fetal kidney has not been reported. This study intends to screen the applicable compound housekeeping genes in rat fetal kidney. In this study, eight housekeeping genes used in kidney studies based on literature reports (GAPDH, ACTB, 18S, HPRT, YWHAZ, HMBS, PPIA, and TBP) were selected as the research object. Their expression levels in the rat fetal kidney in physiological condition and the intrauterine growth retardation (IUGR) model induced by prenatal dexamethasone exposure (PDE) (0.2 mg/kg·day from gestation Days 9 to 20) was measured. Furthermore, these eight housekeeping genes were used to conduct relative quantitative analysis of nephrin expression in the fetal kidney in PDE-induced IUGR model, to compare the influence of choosing different housekeeping gene on data analysis of nephrin expression and to verify the reliability of selected compound housekeeping genes. In this study, stable housekeeping genes of fetal kidney tissues in PDE-induced IUGR model were identified: ACTB, GAPDH, TBP, and HMBS for males; ACTB, YWHAZ, and GAPDH for females. Besides, our results suggest that ACTB + GAPDH were the best compound housekeeping genes for normalization analysis in male fetal kidney studies, and ACTB + YWHAZ in females. This study will provide an experimental evidence basis for the selection of housekeeping genes in the RT-qPCR experiment in renal development toxicology-related models.     

The comparison of cultures,widal agglutination test and polymerase chain reaction as a diagnostic tool in typhoid fever

Typhoid fever caused by Salmonella typhi, paratyphi A and B, is an important cause of morbidity and mortality in many developing countries. A rapid and sensitive method for the detection of S. typhi is essential for early diagnosis of typhoid fever and effective therapy. In this study 45 febrile patients who were suspected to have enteric fever were enrolled, and the results of blood cultures, widal agglutination tests and Polymerase Chain Reaction in these cases were evaluated. Group I consisted of 11 patients with diseases other than salmonella infections, group II represented 6 patients with positive cultures, and group III represented 28 patients with negative blood cultures negative but who were clinically suspected cases that had a medical history of using variable antimicrobial agents. Two positive PCR results were present; one of them was in culture positive group (16,6%) and the other was in culture negative group (3,5%). In our study widal agglutination tests and cultures were found not to be helpful in differential dignosis. Although PCR based detection of S. typhi is reported to be a sensitive and specific test for the diagnosis of enteric fever, in our study the benefit of this method in the diagnosis of especially patients who were treated with antimicrobial therapy was not clearly determined. Other methods to increase sensitiviy and specificity to levels such as those of real time PCR should be developed and large-scaled studies should be done in endemic and non-epidemic regions.     

Infliximab and other immunomodulating drugs in patients with inflammatory bowel disease and the risk of serious bacterial infections

Background There remain concerns about the safety of infliximab therapy in patients with inflammatory bowel disease (IBD). Aim To assess the association between the initiation of infliximab and other immunomodulating drugs and the risk of serious bacterial infection in the treatment of IBD. Methods We assembled a cohort study of patients with IBD, including Crohn’s disease (CD) and ulcerative colitis (UC). All patients initiating an immunomodulating drug between January 2001 and April 2006 were identified in British Columbia from linked health care utilization databases. Exposure of interest was initiation of infliximab or corticosteroids compared with initiation of other immunosuppressive agents, including azathioprine, mercaptopurine (MP) and methotrexate (MTX). Outcome of interest was serious bacterial infections requiring hospitalization, including Clostridium difficile. Results Among 10 662 IBD patients, the incidence rate of bacteriaemia ranged from 3.8 per 1000 person‐years (95% confidence interval 2.1–6.2) for other immunosuppressive agents to 7.4 (3.3–19.3) for infliximab with slightly higher rate for serious bacterial infections resulting in an adjusted relative risk 1.4 (0.47–4.24). Clostridium difficile infections occurred in 0/1000 (0–5.4) among 521 infliximab initiations and 14/1000 (10.6–18.2) for corticosteroids. Corticosteroid initiation tripled the risk of C. difficile infections (RR = 3.4; 1.9–6.1) compared with other immunosuppressant agents. This corticosteroid effect was neither dose‐dependent nor duration‐dependent. Bacteriaemia and other serious bacterial infections were not increased by corticosteroids or infliximab (5 events). Conclusions In a population‐based cohort of patients with IBD, we found no meaningful association between infliximab and serious bacterial infections, although some subgroups had few events. Corticosteroid initiation increased the risk for C. difficile infections in these patients.     

多重PCR技术检测沙门菌两种四环素耐药基因

应用两对特异性引物同时检测四环素耐药基因tetB和tetC通过对35株沙门菌分离株的四环素耐药性检测，表明所有沙门菌分离株均含tetC基因，与药敏试验结果阳性符合率65．7％，8株同时含有tetB基因，与药敏试验结果阳性符合率100％，tetB基因和tetC基因双阳性的菌株与药敏试验结果阳性符合率也为100％。取其中部分菌株扩增出tetB和tetC基因片段进行序列分析，5株菌的tetB基因扩增产物序列完全相同，与质粒pRT11相应序列同源性达99．7％；14株菌的tetC基因扩增产物与质粒pBR322中的相应序列同源性为100％。证实沙门菌耐药基因普遍存在，且同时含有tetB和tetC基因的菌株表现耐药。多重PCR技术同时检测两种四环素耐药基因，适合大量样本的检测，对开展沙门菌四环素多种耐药基因的分子流行病学监测提供了有效途径。     

荧光定量PCR技术检测乳腺癌患者外周血CEA、CK19mRNA表达的意义

目的探讨荧光定量PCR(FQ-PCR)检测外周血癌胚抗原(CEA，eareinoembryonic antigen)、细胞角蛋白19(CK19，cytokeratin 19)m1RNA的表达在乳腺癌外周血微转移诊断中的应用。方法采用FQPCR法，并以β-actin为内对照，测定19例乳腺癌组织中特异性mRNA表达情况，以及测定28例健康女性体检者、11例良性乳腺疾病患者和51例乳腺癌患者的外周血中CEA、CK19mRNA的表达。结果19例乳腺癌组织中CEAmRNA的阳性率为78．9％(15／19)，CK19mRNA的表达阳性率为100％(19／19)。本研究应用FQ-PCR检测外周血CEA、CK19mRNA的表达来检测乳腺癌外周血微转移，以上有一项为阳性即判定为转移。正常对照组和良性乳腺疾病组CEA、CK19mRNA的表达均为阴性，乳腺癌组显著高于前2组。结论FQ-PCR技术是高度灵敏、高度特异的快速定量检测CEA、CK19mRNA的方法，可有助于乳腺癌的诊断和评估预后。