Performance evaluation of two multiplex qualitative RT-PCR assays for detection of respiratory infection in paediatric population.

INTRODUCTION: Acute respiratory infection (ARI) contributes to significant mortality and morbidity worldwide and is usually caused by a wide range of respiratory pathogens. This study aims to describe the performance of QIAstat-Dx® Respiratory Panel V2 (RP) and RespiFinder® 2SMART assays for respiratory pathogens detection. MATERIALS AND METHODS: A total of 110 nasopharyngeal swabs (NPS) were collected from children aged one month to 12 years old who were admitted with ARI in UKMMC during a one-year period. The two qPCR assays were conducted in parallel. RESULTS: Ninety-seven samples (88.2%) were positive by QIAstat-Dx RP and 86 (78.2%) by RespiFinder assay. The overall agreement on both assays was substantial (kappa value: 0.769) with excellent concordance rate of 96.95%. Using both assays, hRV/EV, INF A/H1N1 and RSV were the most common pathogens detected. Influenza A/H1N1 infection was significantly seen higher in older children (age group > 60 months old) (53.3%, p-value < 0.05). Meanwhile, RSV and hRV/EV infection were seen among below one-year-old children. Co-infections by two to four pathogens were detected in 17 (17.5%) samples by QIAstat-Dx RP and 12 (14%) samples by RespiFinder, mainly involving hRV/EV. Bacterial detection was observed only in 5 (4.5%) and 6 (5.4%) samples by QIAstat-Dx RP and RespiFinder, respectively, with Mycoplasma pneumoniae the most common detected. CONCLUSION: The overall performance of the two qPCR assays was comparable and showed excellent agreement. Both detected various clinically important respiratory pathogens in a single test with simultaneous multiple infection detection. The use of qPCR as a routine diagnostic test can improve diagnosis and management.

[Clinical experience of REcanalisation and balloon-oriented puncture for Re-insertion of long- term dialysis catheter in nonpatent central veins].

It is difficult to insert long-term dialysis catheters after severe stenosis or occlusion of the internal jugular vein and innominate vein. We used REcanalisation and balloon-oriented puncture for Re-insertion of dialysis catheter in nonpatent central veins (REBORN) in seven patients with severe central venous lesions, and all patients were inserted with long-term dialysis catheters successfully. None had severe complications such as pneumothorax, hemothorax, or pulmonary embolism during operation. All catheters functioned well after postoperative follow-up of 2 months. REBORN provides a novel approach to establish difficult dialysis pathways.

[Effects of curcumin on neurobehavior and oxidative stress in hippocampus of rats exposed to manganese].

Objective: To establish the animal model of subchronic manganism, and to explore the effect of manganese on neurofunction of rats and the protective effect of curcumin on neurotoxicity of manganism rats. Methods: From July to December 2019, 80 SPF male SD rats were divided into 8 groups according to body weight by random number table method, which were blank control group, low, middle and high dose manganese exposure group, low, middle and high dose curcumin antagonistic group and curcumin group, with 10 rats in each group. The low, middle and high dose manganese groups were given intraperitoneal injection of 5 mg/kg, 10 mg/kg and 15 mg/kg MnCl(2)·4H(2)O respectively. The low, middle and high dose curcumin antagonistic groups were given 100 mg/kg, 200 mg/kg and 400 mg/kg curcumin orally along with 15 mg/kg MnCl(2)·4H(2)O intraperitoneal injection. Curcumin group was given 400 mg/kg curcumin orally. The rats were exposed to 5 days a week, once a day for 16 weeks. After exposure, neurobehavioral tests (balance beam test, Morris water maze, passive avoidance test) were carried out in each group. Hippocampus tissues were taken for pathological examination and oxidative stress indexes were detected. Results: The balance beam test results showed that, compared with the blank control group, the scores of balance beam of the rats in the middle and high dose manganese exposure groups increased (P<0.05) . Compared with the high dose manganese exposure group, the balance beam scores of the low, middle and high dose curcumin antagonistic groups were decreased (P<0.05) .The results of Morris water maze showed that, compared with the blank control group, the escape latency of middle and high dose manganese exposure groups was prolonged from the third day (P<0.05) , and the average number of crossing the platform area of each manganese exposure group was decreased (P<0.05) .Compared with the high dose manganese exposure group, the escape latency of the middle and high dose curcumin antagonistic groups was shortened (P<0.05) , and the average number of crossing the original platform was increased (P<0.05) . The results of passive avoidance test show that, compared with the blank control group, the number of errors were increased in middle and high dose manganese exposure groups (P<0.05) . Compared with the high dose manganese exposure group, the number of errors in the passive avoidance test in the middle and high dose curcumin antagonistic groups were decreased (P<0.05) . Pathological examination showed that the rats treated with manganses had different degrees of degeneration and necrosis of nerve cells, and the structure of nerve cells was blurred and the number of nerve cells decreased. The above phenomena were improved after curcumin antagonism. The results of oxidative stress index showed that, compared with blank control group, the activity of superoxide dismutase (SOD) decreased and the content of malondialdehyde (MDA) increased in the hippocampus of rats exposed to middle and high dose of manganese (P<0.05) . Compared with the high dose manganese exposure group, the SOD activity increased and the MDA content decreased in the middle and high dose antagonist group (P<0.05) . Conclusion: Subchronic manganese exposure can reduce the balance function, learning and memory ability of rats, and damage the hippocampal nerve cells in oxidative stress state. Curcumin can improve the balance function and learning and memory ability of rats with manganese poisoning, improve the hippocampal nerve damage caused by manganese exposure, and has a certain protective effect on manganese induced neurotoxicity.

Analysis of the gene-protein interaction network in glioma.

Glioma is the most aggressive type of brain tumor. Great progress has been achieved in glioma treatment, but the protein-protein interaction networks underlining glioma are poorly understood. We identified the protein-protein interaction network for glioma based on gene expression and predicted biological pathways underlying the molecular complexes in the network. Genes involved in glioma were selected from the Online Mendelian Inheritance in Man (OMIM) database. A literature search was performed using the Agilent Literature Search plugin, and Cytoscape was used to establish a protein-protein interaction network. The molecular complexes in the network were detected using the Clusterviz plugin, and pathway enrichment of molecular complexes was performed using DAVID online. There were 378 glioma genes in the OMIM database. The protein-protein interaction network in glioma contained 1814 nodes, 6471 edges, and 8 molecular complexes. There were 17 pathways (false discovery rate <1), which were related to cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway, chemokine signaling pathway, oocyte meiosis, progesterone-mediated oocyte maturation, transmembrane transport of small molecules, metabolism of amino acids, and notch signaling pathway, among others. Our results provide a bioinformatic foundation for further studies of the mechanisms of glioma.

Role of H3K18ac-regulated nucleotide excision repair-related genes in arsenic-induced DNA damage and repair of HaCaT cells.

Arsenic is an environmental poison and is a grade I human carcinogen that can cause many types of damage to the body. The skin is one of the main target organs of arsenic damage, but the molecular mechanisms underlying arsenic poisoning are not clear. Arsenic is an epigenetic agent. Histone acetylation is one of the earliest covalent modifications to be discovered and is closely related to the occurrence and development of tumors. To investigate the role of acetylated histone H3K18 (H3K18 ac) in arsenic-induced DNA damage, HaCaT cells were exposed to sodium arsenite (NaAsO2) for 24 h. It was found that arsenic induced the downregulation of xeroderma pigmentosum A, D, and F (XPA, XPD, and XPF-nucleotide excision repair (NER)-related genes) expression, as well as histone H3K18 ac expression, and aggravated DNA damage. Chromatin immunoprecipitation quantitative polymerase chain reaction (ChIP-qPCR) analysis showed that H3K18 acetylation in the promoter regions of XPA, XPD, and XPF was downregulated. In addition, the use of the histone deacetylase inhibitor trichostatin A (TSA) partially inhibited arsenic-induced DNA damage, inhibited deacetylation of H3K18 ac in the promoter regions of XPA, XPD, and XPF genes, increased acetylation of H3K18, and promoted the transcriptional expression of NER-related genes. Our study revealed that NaAsO2 induces DNA damage and inhibits the expression of NER-related genes, while TSA increases the H3K18 ac enrichment level and promotes the transcriptional expression of NER, thereby inhibiting DNA damage. These findings provide new ideas for understanding the molecular mechanisms underlying arsenic-induced skin damage.

Parental knowledge, attitudes and antibiotic use for acute upper respiratory tract infection in children attending a primary healthcare clinic in Malaysia.

INTRODUCTION: A study was carried out in a primary healthcare clinic in the Hulu Langat district of Malaysia to assess the parental knowledge, attitudes and antibiotic use for common childhood acute upper respiratory tract infection (URTI). METHODS: A cross-sectional study involving 421 parents, who were surveyed by using an interviewer-administered questionnaire, from April to June 2001. RESULTS: Approximately 59 percent of parents from this study believed that weather was the main cause of acute URTI of their children, 13 percent thought it was due to food, and only about 27 percent said it was caused by germs. Nearly 68 percent, 69 percent and 76 percent of them believed that antibiotics was helpful in treating the common cold, cough and fever, respectively. 29 percent of parents who thought that their child with acute URTI needed antibiotics were not prescribed with any. On the other hand, 17 percent believed that antibiotics were unnecessary when prescribed. 28 percent of parents had requested for antibiotics, and 93 percent received what they requested for their child with acute URTI. About 31 percent of parents who did not request any antibiotics claimed that private general practitioners habitually prescribed antibiotics. The antibiotic compliance was poor with only 74 percent completing the entire course, with 85 percent of them stopping once they improved symptomatically. 15 percent of parents gave "leftover" antibiotics, 24 percent gave "shared" antibiotics, and 5.5 percent bought antibiotics for their child with acute URTI without consulting a doctor. CONCLUSION: This study shows that parents often have inadequate knowledge and misconceptions on antibiotic use for acute URTI in children. Improved parental education may reduce unnecessary antibiotic prescription and antimicrobial resistance in the community.

General pediatricians, pediatric subspecialists, and pediatric primary care.

OBJECTIVE: To assess the respective roles of general pediatricians and pediatric subspecialists in the provision of primary pediatric care. DESIGN AND METHODS: A practice characteristics questionnaire that included questions about primary care was sent to a random sample of 1616 board-certified and board-eligible active Fellows of the American Academy of Pediatrics; 1145 (70.9%) responded. Analyses pertain to those pediatricians who provided ambulatory patient care and were not in graduate medical education training at the time of the survey. Respondents were divided into 2 groups for purposes of analysis: the 527 pediatricians whose practice was primarily in general pediatrics (defined as 80% of time spent in general pediatrics or any time spent in adolescent medicine) and the 213 pediatricians whose practice was subspecialty focused (all others). These groups were then further stratified according to whether they provided primary care. The resultant subgroups contained 518 general pediatricians and 98 subspecialists who provided primary care. RESULTS: Among the entire sample, general pediatricians indicated that general pediatricians provide 93% of the primary care delivered by their practice and that pediatric subspecialists provide 2% of the primary care. In contrast, pediatric subspecialists reported that general pediatricians provide 53% of the primary care delivered by their practice and that subspecialists provide 32% of such care (P<.001). Among the subsample of pediatricians who provide primary care, general pediatricians reported delivering 88% of the primary care received by their patients and subspecialists reported delivering 74% of the primary care received by their patients (P<.001). CONCLUSION: Perspectives on the degree to which pediatric subspecialists provide primary pediatric care vary depending on generalist vs subspecialist self-identification.

Modified fracture brace for tibial fracture with varus angulation: a case report.

Sarmiento introduced the functional fracture brace for the management of tibial shaft fracture in 1963. However, tibial angulation with varus deformity cannot be prevented or corrected by such a device. In this paper, a case of tibial shaft fracture with varus angulation treated with a modified below-knee fracture brace was reported.

Dopa responsive dystonia with diurnal fluctuation (Segawa's syndrome).

A 10-year-old girl presented with progressive dystonia with diurnal fluctuation. Response to low dose L-Dopa was dramatic and sustained with no complications. Recurrence of symptoms was observed on attempted withdrawal. Because of the dramatic response to therapy, dopa-responsive dystonia must be considered in the differential diagnosis of disorders presenting as gait disorders in childhood.

Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen.

The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G(0)-G(1) phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.

The correlation between craniovertebral angle, backpack weights, and disability due to neck pain in adolescents.

PURPOSES: To investigate the responses of the craniovertebral (CV) angle to backpack loadings in adolescents with and without neck pain and to explore the relationships between CV angle, relative backpack weight, neck pain and disability. METHODS: A cross-sectional single-blinded study was conducted on 60 adolescents (30 neck pain and 30 non-neck pain) aged from 13 to 18 years old. The verbal analog scale (VAS) and Chinese version of Northwick Park Neck Pain Questionnaire (NPQ) were used to assess neck pain severity and disability respectively. CV angle was measured in neutral and with backpack loadings of 5% to 30% of subject's body weight by using the Head Posture Spinal Curvature Instrument (HPSCI). RESULTS: In both groups, CV angles gradually decreased with increment of backpack loadings and the amount of decreases became significant from 10% body weight onwards (P < 0.05). Although the changes of CV angles did not show any significant differences at any point of comparison between the groups, the neck pain group showed a clinically significant decrease of CV angle (∼ 5°) at 10% relative loading whereas non-neck pain group did it at 15% relative loading. Change of CV angles did not show significant correlations with relative backpack weight, cervical pain and disability (P > 0.05). CONCLUSIONS: Our findings suggested a safety limit of 10% relative backpack load for adolescents. The results showed the tendency that the ability of maintaining good head posture in response to backpack loadings by non-neck pain subjects might be better than those with neck pain.

The correlation between craniovertebral angle, backpack weights, and disability due to neck pain in adolescents.

PURPOSES: To investigate the responses of the craniovertebral (CV) angle to backpack loadings in adolescents with and without neck pain and to explore the relationships between CV angle, relative backpack weight, neck pain and disability. METHODS: A cross-sectional single-blinded study was conducted on 60 adolescents (30 neck pain and 30 non-neck pain) aged from 13 to 18 years old. The verbal analog scale (VAS) and Chinese version of Northwick Park Neck Pain Questionnaire (NPQ) were used to assess neck pain severity and disability respectively. CV angle was measured in neutral and with backpack loadings of 5% to 30% of subject's body weight by using the Head Posture Spinal Curvature Instrument (HPSCI). RESULTS: In both groups, CV angles gradually decreased with increment of backpack loadings and the amount of decreases became significant from 10% body weight onwards (P < 0.05). Although the changes of CV angles did not show any significant differences at any point of comparison between the groups, the neck pain group showed a clinically significant decrease of CV angle ( approximately 5 degrees ) at 10% relative loading whereas non-neck pain group did it at 15% relative loading. Change of CV angles did not show significant correlations with relative backpack weight, cervical pain and disability (P > 0.05). CONCLUSIONS: Our findings suggested a safety limit of 10% relative backpack load for adolescents. The results showed the tendency that the ability of maintaining good head posture in response to backpack loadings by non-neck pain subjects might be better than those with neck pain.

Spinal cord arteriovenous malformation presenting as chest pain in a child.

STUDY DESIGN: A case report of spinal arteriovenous malformation presented as chest pain in a child. OBJECTIVES: To illustrate the atypical clinical presentation as right chest pain in a child with spinal cord arteriovenous malformation. SETTING: Chang Gung Memorial Hospital and Chang Gung University, Taiwan. CASE REPORT: An 8-year-old boy was diagnosed with spinal cord arteriovenous malformation after paraplegia. However, early diagnosis was delayed for the atypical initial clinical presentation as chest pain. After surgery and rehabilitation for 6 months, the patient could walk without assistance. CONCLUSIONS: Correct diagnosis of spinal cord arteriovenous malformations is sometimes difficult in children with atypical presentations. Early diagnosis relies on detailed examination and high index of suspicion.

Low-dose inhaled nitric oxide for neonates with pulmonary hypertension.

OBJECTIVE: Inhaled nitric oxide (iNO) has been shown to cause selective pulmonary vasodilatation and improve ventilation-perfusion matching and may be an important therapeutic option for the treatment of persistent pulmonary hypertension of the newborn (PPHN). We report our experience on the use of iNO in neonates with severe PPHN. METHODOLOGY: Inhaled NO was administered to 10 infants with PPHN and persistent hypoxaemia (meconium aspiration syndrome, n = 9; pneumonia, n = 1) after failure of conventional therapy to improve oxygenation. With the exception of one infant, iNO was commenced at 10 ppm. RESULTS: After 30 min exposure to iNO, the arterial oxygen tension (PaO2) rose from a median of 49 mmHg (6.5 kPa) [range 12-82 mmHg (1.6-10.9 kPa)] to 75 mmHg (10 kPa) [range 17-450 mmHg (2.3-60 kPa)] (P = 0.005), while the median oxygenation index fell (pre-iNO of 37 vs post-iNO 20) (P = 0.005) and median systemic arterial pressure rose (pre-iNO 46.5 mmHg (6.2 kPa) [range 32-63 mmHg (4.3 to 8.4 kPa vs post-iNO 54.5 mmHg (7.3 kPa) [range 36-74 kPa]) P = 0.005). All infants subsequently continued to receive iNO with the duration of exposure to iNO ranging from 12 to 168 h (median duration 100 h). Three infants died despite showing an initial beneficial response to iNO. The mean duration of intubation for survivors was 11.9 +/- 2.6 days. Methaemoglobinaemia and toxic levels of nitrogen dioxide were not seen during iNO administration. Of the seven survivors, 12 month follow up in two infants and 4 month follow up in four infants showed age-appropriate neurodevelopmental skills, with one infant having very mild hearing loss. CONCLUSIONS: Inhaled NO reduces the oxygenation index by improving the PaO2 and decreasing ventilation pressures, and appears to be clinically useful in severely hypoxaemic infants with PPHN refractory to conventional treatment.

Inhaled nitric oxide during emergency neonatal transportation.

Inhaled nitric oxide is currently being investigated as a selective pulmonary vasodilator for neonates with persistent pulmonary hypertension. The use of continuous inhaled nitric oxide during emergency transportation of three critically III neonates with meconium aspiration and pulmonary hypertension is described. The successful application of this technique may allow safer transportation of neonates who require high level intensive care including ongoing nitric oxide, high frequency ventilation and/or extracorporeal life support. Regionally based nitric oxide-equipped retrieval teams may relieve the pressure on smaller neonatal intensive care units to provide inhaled nitric oxide therapy and allow centralization of nitric oxide resources, thus facilitating development of expertise and the completion of meaningful research programs with substantial recruitment.

Children at risk: their health insurance status by state.

OBJECTIVE: To inform policymakers and child health advocates about children's health insurance coverage in each state as Congress debates proposals to restructure the Medicaid program amidst declining employer-based dependent health insurance coverage. METHODS: Multiple years of data from the March supplement of the Current Population Surveys were pooled to yield more reliable estimates of changes in children's health insurance coverage in each state from 1987 to 1993. RESULTS: Overall, the number of uninsured children grew by nearly 1 million between 1987 and 1993. The proportion of infants and children <7 years old without health insurance declined; the proportion of uninsured children between the ages of 7 and 22 increased. Between 1987-1990 and 1991-1993, six states experienced a significant increase in the number of uninsured children, only two states experienced a significant decrease in the number of uninsured children, although no progress was found in reducing the number of uninsured children in 43 states. Between 1987-1990 and 1991-1993, the proportion of children covered by employer-based private insurance decreased significantly in three-fourths of the states and the proportion of children with Medicaid coverage increased significantly in four-fifths of the states. CONCLUSIONS: In a period of fast-declining employer-based health insurance coverage for dependents, Medicaid expansions have worked to moderate the surge in the number of children without health insurance. Of the 30 million children without private health insurance in 1993, nearly 18 million were insured by Medicaid. As policymakers debate the future of the Medicaid program, they must consider seriously its role as the country's largest insurer of children's health and its efficacy as a vital safety net for the nation's children.

Intracavernous injection of prostaglandin E1 in spinal cord injured patients with erectile dysfunction. A preliminary report.

Our experience with intracavernous injection of prostaglandin E1 in spinal cord injured patients with neurogenic erectile dysfunction included 15 men. They received testing dosage starting from 5 micrograms with increasing dosage (maximum 20 micrograms) to achieve a rigid erection of Schramek's grade 5 and lasting for at least 20 min. All of them had achieved functional erection adequate for coitus after treatment except one patient who had been proved to have venogenic impotence. We found that intracavernous injection of prostaglandin E1 had significantly improved the erectile condition. No systemic side effect or any other complication was noted except that pain at the injection site was complained of in two patients with incomplete lesion.

Randomised trial of three doses of inhaled nitric oxide in acute respiratory distress syndrome.

BACKGROUND: Inhaled nitric oxide (iNO) is a potential therapeutic agent for the management of acute respiratory distress syndrome (ARDS). Concerns remain, however, regarding the potential toxicity from iNO and/or its oxidative derivatives and methaemoglobinaemia. AIMS: To determine the risk of toxicity from iNO, which includes worsening of lung injury, a prospective study evaluating the acute effects of three concentrations of iNO on gas exchange and haemodynamics in 12 children with ARDS was performed in a tertiary paediatric intensive care unit. INTERVENTION: iNO was administered for one hour at three concentrations (1, 10, and 20 parts per million (ppm)) in a random order of possible dosing schedules to avoid dose accumulation bias. Arterial blood gas, methaemoglobin concentrations, and haemodynamic parameters were obtained at baseline before commencement of iNO, at the end of each study hour, and after iNO was discontinued. Nitric oxide and nitrogen dioxide concentrations were continuously monitored during the study. RESULTS: iNO significantly improved the oxygenation ratio (Pao2/Fio2) from a mean (SEM) baseline of 11.9 (1.7) kPa to 20 (3.9) kPa, 24 (4.5) kPa, and 21.6 (3.9) kPa at 1, 10, and 20 ppm iNO, respectively. There was no significant difference in the improvement in oxygenation achieved between the three concentrations. Correspondingly, there was a significant improvement in oxygenation index (pre-iNO 28.3 (5) v post-iNO 18 (3) (1 ppm), 15 (3) (10 ppm), 16 (3) (20 ppm)). No toxicity from methaemoglobinaemia or nitrogen dioxide was seen during iNO administration. CONCLUSION: The results show that a low concentration of iNO (1 ppm) is as effective as higher concentrations (10 and 20 ppm) in improving oxygenation in children with ARDS and may be important in minimising toxicity during iNO use.