BACKGROUND/PURPOSE:Pertussis has been considered a vaccine-preventable "childhood disease", but a shift in age distribution has been reported worldwide. We conducted a seroepidemiological study in 2013 in Taiwan to elucidate the seroprevalence of pertussis among elementary school children.METHODS:With a multilevel randomized method, which included 14 variables (4 population variables, 4 socio-educational variables, and 6 medical facilities' variables), the 29 executive districts of New Taipei City, Taiwan, were categorized into five strata. From each stratum, the number of school children as well as the number of elementary schools were proportionally selected. Enzyme immunoassay was applied for pertussis immunoglobulin-G measurement.RESULTS:A total of 936 children from 14 schools were recruited. Most participants (98.89%) received at least three doses of acellular diphtheria-tetanus-pertussis vaccine. The overall seropositive rate for pertussis was 33.97%. The seropositive rate was highest for students in Grade 1 (49.36%) and then declined with time, except for Grade 6 students. Students from Grade 1 to Grade 4 had a significant higher seropositive rate (37.18% vs. 27.56%, p = 0.002) than those from Grade 5 to Grade 6, but a lower geometric mean titer (18.71 NovaTec Unit/mL vs. 20.04 NovaTec Unit/mL, p = 0.20). For the class grades, geometric mean titers were positively correlated with seroprevalence (p < 0.005).CONCLUSION:Currently, almost one-third of elementary school children in Taiwan were seropositive for pertussis, a rate lower than expected. Seroprevalence declined with increasing class grades except for Grade 6. The current national immunization program may not provide adequate protection for children against pertussis.

BACKGROUND:To establish the seroepidemiologic data of hepatitis A virus (HAV) vaccine-preventable HAV diseases among school children (7-12 years old) attending elementary schools in New Taipei City, Taiwan. This is a pilot study of an ongoing nationwide study, and will be the reference for a national immunization program. METHODS:The school children were selected for samplings, based on a multistage stratified sampling method that included 14 variables (4 socioeducational variables, 4 socioeducational variables, and 6 medical facilities' variables). The 29 administrative districts of New Taipei City were categorized into five strata. In total, 936 school children from 14 schools were recruited and bled for the serologic tests of HAV by enzyme-linked immunosorbent assay method. RESULTS:The seropositive rate for HAV was 8.33% among the 936 children. From each school, the difference in the seropositive rate for HAV ranged 0-18.75%. There was no significant difference between each stratum (p = 0.059) or grade (p = 0.570); however, there was a difference between schools in the first stratum (p = 0.033) that was associated with different vaccination rates. This study also revealed a significantly greater seropositive rate in the vaccination group (p < 0.001) and in females (p = 0.02). CONCLUSION:The seropositive rate for the HAV was <10% and was mostly associated with the vaccination status. Because of the low HAV vaccination rate and low seropositive rate for the HAV, an effective hepatitis A vaccine is a useful tool to prevent HAV infection. It is worthy to discuss whether to include the HAV vaccine as part of a routine vaccination program in Taiwan.

Background: Adenovirus is a common pathogen in the pediatric population. No reports systemically evaluated central nervous system (CNS) dysfunction associated with adenovirus.Objective: To describe the detailed clinical features of adenovirus-associated CNS dysfunction in children. Study design: Of 3298 children with a throat virus culture positive for adenovirus treated at a university-affiliated hospital, from January 2000 to June 2008, in northern Taiwan, medical charts were retrospectively reviewed and those with CNS dysfunction were included.Results: 109 (3.3%) children had signs or symptoms of CNS dysfunction on admission. The median age was 3.6 years and 81 (76%) were less than 5 years of age. 64 (59%) were male. The most frequently cited CNS symptoms were seizure (64%), altered state of consciousness (13%), visual hallucination (9.3%) and lethargy (7.5%). The most common diagnoses included febrile seizure (48%), encephalitis (26%), afebrile seizure (11%) and meningitis/meningismus (8.3%). 31 (46%) of 67 patients receiving electroencephalogram examination had abnormal findings and 13 (36%) of 36 with image studies had abnormal findings. Ninety-nine (91%) children returned to normal health, 7 (6.4%) had a sequel of seizure disorder, and 3 (2.7%) had major sequelae and none died. Nine serotypes were identified, with three major serotypes (types 3, 2 and 1).Conclusion: CNS dysfunction was identified in 3.3% of children with adenoviral infection and most occurred in those <5 years old. The clinical prognosis seemed well. However, CNS dysfunction is a potentially serious complication of adenovirus infection in children. (C) 2013 Elsevier B.V. All rights reserved.

Background: Clinical features of radiologically confirmed pneumonia due to adenovirus in children have not been comprehensively evaluated.Objective: To describe the detailed clinical features of radiologically confirmed adenovirus-associated pneumonia in children.Study design: Of 3298 children with a throat virus culture positive for adenovirus treated at a university-affiliated hospital, from January 2000 to June 2008, in northern Taiwan, 80 hospitalized children (2.4%) with radiologically confirmed pneumonia were identified. From four children with incomplete medical records, only demographics were included for analysis.Results: The median age was 2.97 years, ranging from 25 days to 14 years. Seventy-three patients (96%) had fever, with a median duration of 7 days. The three most common respiratory symptoms were cough (99%), rhinorrhea (82%) and dyspnea (42%). Gastrointestinal symptoms were recorded in 80% of the patients, and neurologic symptoms in four children. Leukocytosis (WBC >= 15,000/mu L) was noted in 19 (25%) patients. Only six patients (8%) had a normal serum C-reactive protein (CRP) value (<5 mg/L), while 48 patients (63%) had a CRP level >40 mg/L. Seventeen (21%) children required intensive care. Seventy-three patients (96%) recovered uneventfully. Sequelae were seen in two patients and death in 1. Of the 69 isolates with serotyping determination, seven serotypes were identified, with a predominant serotype (type 3 for 73%).Conclusion: Less than 5% of the children with adenoviral infection had radiologically confirmed patch pneumonia. The manifestation of pneumonia caused by adenovirus was similar to that of bacterial pneumonia. (C) 2012 Elsevier B.V. All rights reserved.

Human metapneumovirus (hMPV) causes both upper and lower respiratory tract infection worldwide. Because little is known about the estimated level of exposure to hMPV in Taiwan, data for 1999 are presented. One thousand one hundred twenty sera from otherwise healthy community-based subjects aged 1 month to 99 years were tested in an enzyme-linked immunosorbent assay; the proportion of seropositive individuals was 68.1%. Moreover, 53.2% of preschool children were seronegative, whereas 88.3% of school-aged individuals and 93.7% of adults had antibodies to hMPV. In summary, the present study provides an earlier evidence of a high rate of exposure to hMPV characterized the seroprevalence among Taiwanese individuals in 1999. In addition, our findings suggest that preschool children in particular would benefit from vaccination against hMPV.

The susceptibility of five cell lines, RD, MRC-5, MK-2, Hep-2 and A549, for various serotypes of human enteroviruses was evaluated. RD cells were susceptible to most serotypes of enteroviruses, especially for human enterovirus A. Consequently, a high prevalence of human enterovirus A in Taiwan may reflect the relative importance of RD cells in the clinical virology laboratory. However, susceptibilities of RD cells to coxsackievirus A9 and A24, and echovirus 9 and 11 were exceptionally low. Alternatively, MRC-5 or MK-2 cells can improve the isolation of these four enteroviruses.

BACKGROUND:The pandemic (H1N1) 2009 has become a threat of public health. To manage rapidly increased infections and disease control, a simple and reliable first-line screening test for viral infection is on urgent demand. METHODS:Through comprehensive bioinformatics analysis, a single nucleotide polymorphism in nucleoprotein gene which differentiates swine lineage virus and human seasonal H1N1 virus was selected as target. A TaqMan probe-based allele discrimination analysis was designed to analyze clinical samples. In total, 93 clinical specimens and 39 viral isolates were used to test the assay efficacy. Traditional viral culture and molecular analysis were used as gold standard. RESULTS:The testing results showed that the established assay has high sensitivity and specificity (92% and 100%) for pandemic (H1N1) 2009. The assay could detect as low as 5 copies of NP gene of pandemic (H1N1) 2009 or 2 viral particles of human seasonal H1N1. CONCLUSION:This assay can be used as a first-line screening and confirmation test for pandemic (H1N1) 2009 virus and human seasonal flu in one-tube reaction. The assay can serve as a convenient method to reduce the burden of PCR manipulation for diagnostic laboratories when large amount of samples need to be analyzed in a short time.

We conducted a study during an outbreak of coxsackievirus B3 infection in 2005 and found that viral RNA could be detected in patients' blood specimens soon after the onset of fever, and the level of viral RNA was positively correlated with disease severity. Timely diagnosis is possible in severe neonatal enterovirus infection.

BACKGROUND/PURPOSE:Some isolated influenza strains are untypable. The possibility that a novel and potentially lethal strain, such as H5 virus, might be one of these untypable strains remains a major concern. This study investigated the nature of untypable influenza strains in Taiwan. METHODS:Subtyping for unsubtyped isolates was conducted by reverse transcription-polymerase chain reaction (RT-PCR) using different primer sets. Sequence determination and comparison, and characterization of viral growth rate were performed. RESULTS:Analysis of viral re-cultures with the different primer sets for RT-PCR revealed that all isolates were of the H3N2 subtype. All isolates resembled A/Fujian/411/02 (H3N2), and were genetically distinct from N2 of A/chicken/Taiwan/1209/03 (H5N2) on phylogenetic analysis. Real-time RT-PCR quantification of viral loads in the 16 clinical isolates revealed that negative results were partially caused by low RNA levels in the culture fluid. Comparison of replication kinetics in Madin-Darby canine kidney cells of recent influenza A H3N2 isolates showed that some Fujian-like isolates from 2003 to 2004 had significantly lower growth rate than Panama-like isolates from 2002. CONCLUSION:The results of this study suggest that some recent isolates may replicate poorly in tissue culture and fail to be typed by current methodology.

Chromosomal passenger proteins including Aurora B, Survivin, and Borealin/Dasra B, also called CDCA8/FLJ10468, are known to play crucial roles during mitosis and cell division. Inappropriate chromosomal segregation and cell division may cause auneuploidy leading to cancer. However, it is still unclear how the expression of chromosomal passenger proteins may be linked to cancer. In this study, we demonstrated that Borealin is a cell cycle-regulated gene and is upregulated at G2-M phases of the cell cycle. We showed that Borealin interacts with Survivin but not with Aurora B. The interaction domain of Survivin in Borealin was mapped to the N-terminal 92 amino-acid residues of Borealin. To examine the linkage between expression of Borealin and cancer, we performed immunohistochemistry analysis using anti-Borealin specific antibody on the paraffin-embedded gastric cancer tissues. Our results showed that Borealin expression is significantly correlated with Survivin (P = 0.003) and Ki67 (P = 0.007) in gastric cancer. Interestingly, an increased nuclear Borealin level reveals borderline association with a poor survival rate (P = 0.047). Taken together, our results demonstrated that Borealin is a cell cycle-regulated chromosomal passenger protein and its aberrant expression is linked to a poor prognosis for gastric cancer.

BACKGROUND:Human metapneumovirus (hMPV) was first recognized in the Netherlands in 2001 from nasopharyngeal aspirate samples and was associated with respiratory tract illness in the pediatric population. This was the first report of metapneumovirus infections in community-acquired pneumonia in Taiwan.METHODS:A total of 116 nasopharyngeal aspirate samples from patients with community-acquired pneumonia was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Other respiratory tract pathogens were also examined. The clinical characteristics and laboratory data were analyzed.RESULTS:Out of the 116 patients, potential causative agents were detected in 95 (81.9%) patients. A total of six human metapneumovirus RT-PCR positive samples was identified. All of these had evidence of coinfection with bacteria (3 Streptococcus pneumoniae, 2 Mycoplasma pneumoniae, 1 Chlamydia pneumoniae). Coinfection with other respiratory viruses was also observed in two cases (1 influenza A, 1 parainfluenza type 3). The age distribution was seven to 11 years except for one patient who was two years of age (Case 1). The most common clinical findings were fever (6/6, 100%), cough (6/6,100%), rhinorrhea (5/6, 83.3%), rales (5/6, 83.3%) and wheezing (1/6, 16.7%). Chest radiographs revealed four with lobar patches and two with interstitial infiltrations. The mean duration of hospital stay was 5.5 +/- 2.8 days. All patients made a complete recovery.CONCLUSIONS:hMPV was identified in 5.2% of patients with community-acquired pneumonia. Our data showed a high rate of coinfection with hMPV in community-acquired pneumonia. Human metapneumovirus infection, like other respiratory viruses, may predispose to secondary bacterial pneumonia.

Transforming growth factor-beta (TGF-beta) has been shown to induce apoptosis in normal hepatocytes and hepatoma cells both in vivo and in vitro. However, the mechanism by which TGF-beta induces apoptosis is not clear. The antiapoptotic activity of antioxidants including N-acetyl-L-cysteine (Ac-Cys), ascorbic acid and a novel free radical scavenger, carboxyfullerene (C60) on TGF-beta-treated human hepatoma Hep3B cells was examined. Only the water-soluble hexacarboxylic acid derivative of C60 was found to prevent TGF-beta-induced apoptosis. Antiapoptotic activity of C60 correlated its ability to eliminate TGF-beta-generated reactive oxygen species (ROSs). However, C60 did not interfere with TGF-beta-activated PAI-1 promoter activity in the Hep3B cells. These results indicate that the signaling pathway of TGF-beta-induced apoptosis may be related to the generation of ROSs and may be uncoupled from the TGF-beta-activated gene promoter activity. Furthermore, the regioisomer of C60 with a C3 symmetry was more potent in protecting cells from apoptosis than that with a D3 symmetry, and the C3 isomer had stronger interactions with lipid bilayers than the D3 isomer. The spectroscopic analysis revealed that the C3 isomer had stronger interactions with artificial lipid bilayers than the D3 isomer. Therefore, our study indicates that C60 may interact with membrane to eliminate TGF-beta-induced ROSs and to prevent apoptosis occur in human hepatoma cells.