Unambiguous molecular detections with multiple genetic approach for the complicated chromosome 22q11 deletion syndrome.

BACKGROUND: Chromosome 22q11 deletion syndrome (22q11DS) causes a developmental disorder during the embryonic stage, usually because of hemizygous deletions. The clinical pictures of patients with 22q11DS vary because of polymorphisms: on average, approximately 93% of affected individuals have a de novo deletion of 22q11, and the rest have inherited the same deletion from a parent. Methods using multiple genetic markers are thus important for the accurate detection of these microdeletions. METHODS: We studied 12 babies suspected to carry 22q11DS and 18 age-matched healthy controls from unrelated Taiwanese families. We determined genomic variance using microarray-based comparative genomic hybridization (array-CGH), quantitative real-time polymerase chain reaction (qPCR) and multiplex ligation-dependent probe amplification (MLPA). RESULTS: Changes in genomic copy number were significantly associated with clinical manifestations for the classical criteria of 22q11DS using MPLA and qPCR (p < 0.01). An identical deletion was shown in three affected infants by MLPA. These reduced DNA dosages were also obtained partially using array-CGH and confirmed by qPCR but with some differences in deletion size. CONCLUSION: Both MLPA and qPCR could produce a clearly defined range of deleted genomic DNA, whereas there must be a deleted genome that is not distinguishable using MLPA. These data demonstrate that such multiple genetic approaches are necessary for the unambiguous molecular detection of these types of complicated genomic syndromes.

Intracellular acidification enhances neutrophil phagocytosis in chronic haemodialysis patients: possible role of CD11b/CD18.

BACKGROUND: We have demonstrated that uraemic neutrophils that exhibit a low intracellular pH (pHi) display enhanced phagocytosis. However, the underlying cellular mechanism is unclear. METHODS: We used neutrophils from three groups of haemodialysis (HD) patients before dialysis (Groups A, B and C) and also from age- and sex-matched healthy individuals to determine pHi, phagocytosis and expression of CD11b, CD18, CD14 and toll-like receptors (TLR)-2 and TLR-4. The patients were categorized based on three consecutive monthly pre-dialysis plasma bicarbonate concentrations(P(HCO3)) and pH values; Groups A, B and C had a constant pre-dialysis P(HCO3) of /=26 mmol/L (mEq/L), respectively. We also studied the effects induced by the correction of metabolic acidosis and monoclonal antibodies (mAbs) against CD11b/CD18 on neutrophils in Group A. Furthermore, we investigated the effect of intracellular acidification on uraemic neutrophils ex vivo. RESULTS: We observed that the neutrophils in Group A exhibited significantly increased phagocytosis and expression of CD11b/CD18 compared with those in Groups B and C. Additionally, our ex vivo studies demonstrated that the mAbs against CD11b/CD18 partially blocked the enhancement of neutrophil phagocytosis in Group A. Moreover, the pHi of uraemic neutrophils is inversely correlated with phagocytosis and expression of CD11b/CD18. CONCLUSIONS: HD patients with a low P(HCO3) exhibited low neutrophil pHi that in turn increased the expression of CD11b/CD18 compared with neutrophils with a normal or high pHi. This increased expression of CD11b/CD18 on the uraemic neutrophils may contribute to the pHi-mediated phagocytosis.

Is C-reactive protein a prognostic factor of colorectal cancer?

PURPOSE: C-reactive protein, a commonly used inflammation marker, has been reported to be a prognostic factor of colorectal cancer. This prospective study was designed to confirm the prognostic value of its preoperative levels and to observe their perioperative change. METHODS: Between January 2001 and September 2005, preoperative C-reactive protein levels were obtained for 212 consecutive patients (140 males) receiving elective open resection of colorectal cancer. A level higher than 0.5 mg/dl was defined as positive. They were analyzed against clinicopathologic factors. The survival of 158 curative resections was analyzed. Postoperative levels (at months 1, 3, and 6) were collected for analysis of changing trend, from the patients receiving curative surgeries. RESULTS: Median value of preoperative C-reactive protein was 0.54 mg/dl (48.6 percent positive). Positive rate was significantly correlated with ulcerative type, larger size, higher stage, and positive carcinoembryonic antigen (>5 ng/ml). In both univariate log-rank test and multiple Cox proportional hazards regression, stage (univariate P = 0.011, and multivariate P = 0.016; hazard ratio, 6.23; 95 percent confidence interval, 1.41-27.54), C-reactive protein (0.5 mg/dl; P = 0.005, and P = 0.016; hazard ratio: 6.51; 95 percent confidence interval: 1.41-30.05), and differentiation (P = 0.006, and P = 0.043; hazard ratio, 3.53; 95 percent confidence interval, 1.04-11.98) were significant factors. Analysis of disease-free interval showed C-reactive protein was significant (P = 0.03): as level rose, prognosis worsened. The quiescent inflammation-response group (< or =0.1 mg/dl) had excellent outcomes. Postoperatively, the C-reactive protein levels declined at the third postoperative month. CONCLUSIONS: Preoperative C-reactive protein is an independent prognostic factor. The levels declined postoperatively, although with a lag. These findings seem to support the response hypothesis regarding C-reactive protein.