1.Stretch-induced Expression of CYR61 Increases the Secretion of IL-8 in A549 Cells via the NF-κβ/Iκβ Pathway
Yan ZHANG ; Ping GUI ; Shang-Long YAO ; Dong YANG ; Yang LV ; De-Fang DING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(4):672-678
Mechanical ventilation (MV) with large tidal volumes can increase lung alveolar permeability and initiate inflammatory responses,resulting in ventilator-induced lung injury (VILI).The mechanisms of the injurious effects of MV and the genetic susceptibility remain unclear.VILI-related genes such as cysteine-rich angiogenic inducer 61 (Cyr61)have been demonstrated to play a detrimental role in the aggressive ventilation strategies.In the present study,we investigated the involvement of Cyr61 in the VIM and the underlying mechanism.A549 cells were exposed to cyclic stretch of varying durations and then the mRNA and protein levels of Cyr61 were measured by real-time PCR and Western blotting,respectively.Additionally,after exposure ofA549 cells to cyclic stretch for 5 min to 1 h,the expression levels of nuclear factor kappaB (NF-κB) and IL-8 were detected by ELISA and Western blotting.Thereafter,Cyr61 expression was depressed in A549 cells with the siRNA pGenesill.1-Cyr61-3 before the cyclic stretch,and IL-8 secretion and the activation of NF-κB pathways were probed by ELISA and Western blotting,respectively.Moreover,a NF-κB inhibitor (PDTC) and an activator (TNF) were used before mechanical stretch.Realtime PCR and ELISA were performed to detect the mRNA and protein of IL-8,respectively.The results showed that the mechanical cyclic stretch led to increased Cyr61 expression at mRNA and protein levels in A549 cells.Additionally,cyclic stretch also mobilized NF-κB from the cytoplasm to the nucleus and increased IL-8 secretion in A549 cells.The inhibition of Cyr61 blocked the NF-κB activation and IL-8 secretion in response to cyclic stretch.Inhibition of NF-κB attenuated the mRNA and protein expression of IL-8 in A549 cells transfected with Cyr61 siRNA.It was suggested that Cyr61/NF-κB signaling pathway mediates the upregulation of IL-8 in response to cyclic stretch in A594 cells.These findings support the hypothesis that Cyr61 plays a critical role in acute lung inflammation triggered by mechanical strain.
2.Construction of small interfering RNA targeting heparanase gene and its inhibitory effect on the in-vasiveness of human malignant melanoma cell line A375 in vitro
Xiao-Yan LIU ; Hong FANG ; Zheng-Gang YANG ; Li-Ming RUAN ; De-Ren FANG ; Ying-Guo DING ; Yi-Na WANG ; Yu ZHANG ; Xiao-ling JIANG ;
Chinese Journal of Dermatology 2003;0(11):-
Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P
3.Effects of ICAM-1 gene K469E, K56M polymorphisms on plasma sICAM-1 expression levels in Chinese Yugur, Tibetan and Han nationalities.
Ming-Ying WANG ; De-Cheng BAI ; Ping ZHU ; Yu FU ; Ding-Fang BU ; Ying ZHANG
Journal of Experimental Hematology 2012;20(5):1205-1211
This study was purposed to investigate the intercellular cell adhesion molecule-1 (ICAM-1) gene K469E (A/G) (rs5498) and K56M (A/T) (rs5491) single nucleotide polymorphisms (SNP) and soluble ICAM-1 (sICAM-1) levels in plasma in three Chinese populations of Yugur, Tibetan and Han nationalities, to analyze comparatively the genotypes and allele frequencies distribution in different ethnic groups, and to explore the effects of ICAM-1 K469E and K56M polymorphism and sICAM-1 levels in plasma. EDTA-anticoagulant venous blood from Yugur(327 cases), Tibetan (400 cases) and Han (126 cases) people was collected, the DNA was extracted by using whole blood genomic DNA extraction kit, DNA SNP were analyzed by PCR-RFLP, genotype was judged by gel scan imaging system after agarose gel electrophoresis, the gene sequence was determined and the distribution of ICAM-1 genotypes and allele frequencies were compared among different ethnic groups, besides, the group representativeness was tested via the Hardy-Weinberg genetic equilibrium. Finally, the human sICAM-1 plasma levels were detected by using human ICAM-1 ELISA kit. The results showed that DNA sequencing result was consistent with PCR-RFLP analysis. In Yugur, Tibetan and Han nationalities, the KK, KE and EE three genotypes at ICAM-1 K469E gene locus were detected, the genotype distribution was not statistically significantly different, while the K, E allele frequency distribution was statistically significantly different (P < 0.05). Both of genotype and allele frequency distribution between Yugur, Tibetan and Han nationalities were statistically significantly different (P < 0.05). In K56M site only KK, KM two genotypes were detected, but the MM genotype was not detected in the three ethnic groups; the difference of two genotypes and K, M allele frequencies between Yugur and Han population was statistically significantly different (P < 0.05). Among three ethnic groups, the sex ratio and age distribution of K469E, K56M genotypes and allele frequencies of ICAM-1 gene were not significantly different, and distribution was in accordance with Hardy-Weinberg genetic equilibrium (P > 0.05). The plasma sICAM-1 level at ICAM-1 K469E allele locus in K individuals [(253 ± 122), (185 ± 97) µg/L] was higher than that at non-K allele [(145 ± 110) µg/L, P < 0.01]; the plasma sICAM-1 level of ICAM-1 K56M sites with KK genotype [(253 ± 122) µg/L] was higher than that of the KM genotypes [(168 ± 103) µg/L, P < 0.01]. In Yugur and Tibetan groups, the plasma sICAM-1 levels [(224 ± 80), (214 ± 111) µg/L] were higher than that in the Han group [(175 ± 125)µg/L, P < 0.05]. Pairwise comparison indicated that the plasma sICAM-1 levels between Yugur and Han group were statistically significantly different (P < 0.01), that was significantly different between Tibetan and Han group (P < 0.05). It is concluded that in Yugur, Tibetan and Han population, the genotypes and gene frequencies of two amino acid sites K469E and K56M in ICAM-1 were KK/KE-type, KK-type and K allele, moreover, the ratio of them in Yugur and Tibetan group was higher than that in Han, while there is not significant difference in sex ratio and age distribution, therefore, ICAM-1 genotype and allele frequency distribution in this study had ethnic representativeness. ICAM-1 gene K469E and K56M polymorphisms were likely to affect the plasma sICAM-1 expression level. K469E gene K allele may be a genetic risk factor, while K56M gene M allele a may be genetic protective factor for some diseases.
Adolescent
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Adult
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Aged
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Asian Continental Ancestry Group
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genetics
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Ethnic Groups
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genetics
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Female
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Gene Frequency
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Genotype
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Humans
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Intercellular Adhesion Molecule-1
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blood
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genetics
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Male
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Middle Aged
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Plasma
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metabolism
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Polymorphism, Genetic
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Young Adult
4.A1381T and -1793G/C polymorphisms of vWF gene impact the plasma vWF levels in Yugur, Tibetan and Han nationalities of China.
Yu FU ; Ping ZHU ; Ding-Fang BU ; Ying ZHANG ; Ming-Ying WANG ; Yi-Wen GONG ; De-Cheng BAI
Journal of Experimental Hematology 2012;20(5):1200-1204
The aim of this study was to investigate the similarities and differences of A1381T (rs216311) and -1793G/C (rs7966230) single nucleotide polymorphisms (SNP) in Chinese Yugur, Tibetan, and Han nationalities and their influence on plasma vWF concentration in order to explore the sensitivity of these 3 nationalities to vWF-related diseases. Peripheral venous blood was obtained from 322 Yugur, 399 Tibetan, and 120 Han healthy people. The DNA were then extracted. vWF gene A1381T and -1793G/C polymorphisms were analyzed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequenced when it was necessary. The vWF:Ag level in plasma was determined by ELISA. The results showed that the genotype distribution of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han nationalities was different with statistically significance (P < 0.05). GG genotype of A1381T locus accounted for 69.9% in Yugur nationality, which was much higher than 56.6% and 53.3% in Tibetan and Han nationalities respectively(P < 0.01); AA genotype of A1381T locus expressed a low level of vWF in plasma. For the -1793G/C locus, the proportion of CG genotype in Yugur was much higher than that in Han, CC genotype expressed a high level of vWF in plasma. The plasma vWF levels with different nationalities and the polymorphism of vWF gene were significantly different. It is concluded that the polymorphisms of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han are significantly different; the polymorphism of vWF gene influences the plasma vWF level; the plasma vWF levels in Yugur and Tibetan are significantly higher than that in Han, which may be associated with the living environment and habits.
Adolescent
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Adult
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Aged
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Asian Continental Ancestry Group
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genetics
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China
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Ethnic Groups
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genetics
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Female
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Gene Frequency
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Genotype
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Humans
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Male
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Middle Aged
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Plasma
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chemistry
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Polymorphism, Genetic
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Young Adult
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von Willebrand Factor
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genetics
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metabolism
5.Effectiveness and safety of different doses of pioglitazone in psoriasis: a meta-analysis of randomized controlled trials
Zhang JING-ZHAN ; Ding YUAN ; Xiang FANG ; Yu SHI-RONG ; Zhang DE-ZHI ; Guan MENG-MENG ; Kang XIAO-JING
Chinese Medical Journal 2020;133(4):444-451
Background:Pioglitazone may be beneficial in the treatment of psoriasis.However,based on the effectiveness and safety considerations,it has not been widely used.To fully evaluate the strength of evidence supporting psoriasis treatment with pioglitazone,we conducted a meta-analysis of existing published studies.Methods:PubMed,Ovid,Cochrane Library,Google Scholar,and Web of Science databases were systematically searched before February 2019.Randomized controlled trials (RCTs) of pioglitazone administration compared with placebo,administered to patients with psoriasis for at least 10 weeks,and published in English were included.Quality of the included RCTs was identified by the modified Jadad scale.The quality of evidence for each outcome was evaluated using the GRADEpro Guideline Development Tool online software.Primary outcomes were proportion of patents showing psoriasis area and severity index (PASI) score improvement (>75%) and the mean percent change in PASI score from baseline to the end of treatment.Dichotomous data were analyzed using odds ratios (ORs) corresponding to the 95% confidence interval (CI),whereas continuous variables,expressed as mean and standard deviation,were analyzed using the mean differences (MD) with the 95% CI.Results:Six RCTs were analyzed.Meta-analysis showed that pioglitazone reduced the PASI scores in patients with psoriasis compared with the control group when administered at 30 mg per day (P < 0.001,MD =-3.82,95% CI =-5.70,-1.93) and at 15 mg per day (P =0.04,MD =-3.53,95% CI =-6.86,-0.20).The PASI-75 of the pioglitazone group was significantly higher than that of the control group at 30 mg per day (P < 0.001,OR =8.30,95% CI =3.99,17.27) and at 15 mg per day (P =0.03,OR =2.96,95% CI =1.08,8.06).No statistically significant differences in total adverse events were observed between the groups.There were no significant differences in common adverse reactions such as weight gain and elevated liver enzymes between the two pioglitazone groups.Conclusions:Use of pioglitazone in the current treatment of psoriasis is beneficial.The therapeutic effect of the daily 30 mg dose may be greater than that of the 15 mg dose per day with no significant change in the frequency of adverse reactions.
6.Detection and clinical study on coronavirus HKU1 with acute lower respiratory tract infections of hospitalized children in Changsha
Ni-Guang XIAO ; Zhi-Ping XIE ; Qiong-Hua ZHOU ; Rong-Fang ZHANG ; Li-Li ZHONG ; Han-Chun GAO ; Xiao-Fang DING ; Jia LI ; Jing-Rong SONG ; Yun-De HOU ; Bing ZHANG ; Zhao-Jun DUAN
Chinese Journal of Experimental and Clinical Virology 2011;25(1):2-4
Objective In order to understand the epidemiological and virologic characteristics of coronavirus HKU1 infection in hospitalized children with acute respiratory tract infection (ARTI) in Changsha. Methods 1165 nasopharyngeal aspirates (NPA) specimens were collected from hospitalized children with ARTI between September 2007 and August 2008 in Changsha. Specimens were screened for pol gene of coronavirus HKU1 by polymerase chain reaction. All positive amplification products were confirmed by sequencing and compared with those in GenBank. Results Coronavirus HKU1 were detected in 12patients (1.03%) out of the 1165 children. The patients were from 8 days to 3 years. The most common clinical diagnosis was bronchopneumonia( 83. 33% ). Similarity of coronavirus HKU1 with those published in the GenBank at nucleotide levels was 98. 18%-100%. Conclusion Coronavirus HKU1 may be important pathogens in children with acute lower respiratory tract infection. Coronavirus HKU1 infections are common in children under 3 years old. There is no significant difference in the infectious rate between the boys and the girls. The peak of its prevalence is in spring and winter. A single genetic lineage of Coronavirus HKU1was revealed in human subjects in Changsha.
7.Viral etiology of 1165 hospitalized children with acute lower respiratory tract infection.
Ni-Guang XIAO ; Bing ZHANG ; Zhao-Jun DUAN ; Zhi-Ping XIE ; Qiong-Hua ZHOU ; Li-Li ZHONG ; Han-Chun GAO ; Xiao-Fang DING ; Sai-Zhen ZENG ; Han HUANG ; Yun-De HOU
Chinese Journal of Contemporary Pediatrics 2012;14(1):28-32
OBJECTIVETo explore the viral etiology of acute low respiratory tract infection (ALRTI) among hospitalized children in Changsha of Hunan Province of China.
METHODSNasopharyngeal aspirates were collected from 1165 hospitalized children with ALRTI in Changsha from September 2007 to August 2008. Respiratory syncytin virus (RSV), human rhinovirus (HRV), influenza virus A (IFVA), influenza virus B (IFVB), parainfluenza 1-3 (PIV 1-3), human metapneumovirus (hMPV), human coronaviruses NL63 (HCoV-NL63), and human coronaviruses HKU1 (HCoV-HKU1) were detected by reverse transcription polymerase chain reaction (RT-PCR). Adenovirus (ADV) and human bocavirus (HBoV) were detected by standard polymerase chain reaction (PCR). WU polyomaviruses (WUPyV) and KI polyomaviruses(KIPyV) were detected by nested PCR. The positive samples further underwent genetic sequencing.
RESULTSAmong the 1165 nasopharyngeal aspirates, viruses were detected in 871 samples (74.76%), among which RSV (27.03%) was the most common virus, followed by HRV (17.33%), PIV3 (13.73%), HBoV (8.67%) and hMPV (6.52%). The overall positive rate of viral detection showed no significant differences between males and females (X2=2.241, P=0.134), whereas the positive rates of PIV3, hMPV, and HBoV in males were higher than in females. The positive rate of viral detection showed significant differences among different age groups (X2=10.934, P=0.027), and the highest positive rate was noted in the age group of 6 months to 1 year. Furthermore, the overall positive rate of viral detection showed a significant difference in term of seasonal distribution, with a peak prevalence in winter.
CONCLUSIONSVirues predominate in the etiology of pediatric ALRTI in Changsha, and RSV, HRV and PIV3 are the main viruses for ALRTI. HBoV and hMPV have become increasingly important. Viral infection-associated ALRTI shows a prevail in the age group of 6 months to 1 year as well as in winter.
Adolescent ; Age Distribution ; Child ; Child, Hospitalized ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Nasopharynx ; virology ; Respiratory Tract Infections ; etiology ; virology ; Seasons ; Sex Distribution ; Viruses ; isolation & purification
8.Long distance-PCR for detection of factor VIII gene inversion in patients with severe hemophilia A.
Pei-Fang DING ; Wei-Sheng SUN ; Qin-You WANG ; De-Chun LIU ; Xue-Qin ZHANG ; Bin TENG ; Fa-Kui SHEN
Journal of Experimental Hematology 2003;11(4):390-392
The aim of current study was to detect intron 22 inversion of factor VIII gene in severe hemophilia A (HA) patients and screen the carriers of the gene inversion. Fifty-five cases of severe HA were involved and factor VIII gene inversion was detected and identified by long distance-PCR (LD-PCR) and 0.6% agarose gel electrophoresis. The 11 kb and 12 kb bands indicate the factor VIII gene inversion and non-inversion, respectively. Occurring of both 11 kb and 12 kb bands indicates a carrier of the inversion. The results showed that factor VIII gene inversion existed in 22 out of 55 cases, which accounted for about 40% of total detected patients. Five carriers of factor VIII gene inversion were diagnosed from the members in 15 families. In conclusion, LD-PCR assay is a simple, rapid and accurate method for detection of factor VIII gene inversion, and this approach is helpful in screening, carrier testing, and prenatal diagnosis of severe hemophilia A.
Adolescent
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Adult
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Antigens
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analysis
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Child
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Child, Preschool
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Chromosome Inversion
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Factor VIII
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genetics
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Hemophilia A
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blood
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genetics
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Humans
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Infant
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Male
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Polymerase Chain Reaction
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methods
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von Willebrand Factor
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immunology
9.Inductively coupled plasma mass spectrometry for the simultaneous determination of thirty metals and metalloids elements in blood samples.
Chun-guang DING ; Chun ZHU ; De-ye LIU ; Ming DONG ; Ai-hua ZHANG ; Ya-juan PAN ; Hui-fang YAN
Chinese Journal of Preventive Medicine 2012;46(8):745-749
OBJECTIVETo establish an inductively coupled plasma mass spectrometry(ICP-MS) method for determination of 30 trace elements including As, Ba, Be, Bi, Ni, Cd, Co, Cr, Cs, Cu, Ga, Mn, Pb, Sr, Tl, V, Ge, Mo, Nb, Ti, W, Te, Se, Zr, In, Sb, Hg, Ce, La, and Sm in human blood.
METHODThe blood samples were analyzed by ICP-MS after diluted 1/10 with 0.01% Triton-X-100 and 0.5% nitric acid solution. Y, Rh and Lu were selected as internal standard in order to correct the matrix interference of Cr, As, Se, and Hg by a hex pole-based collision-reaction cell. Other elements were determined with standard method. The limits of detection, precision and accuracy of the method were evaluated. The accuracy was validated by the determination of the whole blood reference material.
RESULTSAll the 30 trace elements have good linearity in their determination range, with the correlation coefficient > 0.9999. The limits of detection of the 30 trace elements were in the range of 1.19 - 2.15 µg/L and the intra-precision and inter-precision (relative standard deviation, RSD) were less than 14.3% (except Hg RSD < 21.2%, and Ni RSD < 15.4%). The spiked recovery for all elements fell within 59.3% - 119.2%. Among the 13 whole blood reference materials, V, Cr, Mn, Co, Ni, Cu, As, Se, Cd, Te, and Pb (1.45, 1.19, 18.40, 0.18, 1.57, 591.00, 2.97, 61.00, 0.35, 1.86, and 9.70 µg/L respectively) fell within the acceptable range and the detection results of Hg (0.59 µg/L) and Mo (1.59 µg/L) were slightly beyond the range.
CONCLUSIONThis method was simple, fast and effective. It can be used to monitor the multi-elementary concentration in human blood.
Humans ; Limit of Detection ; Mass Spectrometry ; methods ; Metalloids ; blood ; Metals ; blood ; Trace Elements ; blood
10.Stroke literacy in Singapore: data from a survey of public housing estate residents.
Weiyen LIM ; Ding Fang CHUANG ; Koy Min CHUE ; Damian Z LEE ; Nicholas J LEONG ; Zhi Guang NG ; Kailing PENG ; Xiayan SHEN ; Ye Ni THAM ; Kangjie WANG ; Deidre Anne De SILVA
Annals of the Academy of Medicine, Singapore 2014;43(9):454-463
INTRODUCTIONKnowledge of stroke symptoms is associated with seeking medical attention early, and knowledge of risk factors is an essential factor in stroke prevention. In this study, we evaluated the level of stroke literacy in Singapore.
MATERIALS AND METHODSA cross-sectional study of Singapore citizens and permanent residents aged 21 years and above was conducted in a public housing estate. Participants were randomly sampled using multi-stage stratified sampling. Assessment of awareness of stroke symptoms and risk factors was performed using open-ended questions. In total, 687 respondents were recruited, with a response rate of 69.7%.
RESULTSOverall, 52.4% of respondents identified the brain as the source of pathology, and 47.6% could cite at least 1 of the 3 FAST symptoms (facial droop, arm weakness and speech difficulty), while 40% could name 2 or more of 7 established risk factors for stroke (high blood pressure, high cholesterol, cigarette smoking, diabetes mellitus, older age, previous heart attacks and stroke). Respondents at higher risk of stroke (older individuals and those with stroke risk factors) did not have greater awareness of stroke symptoms and risk factors. The majority of respondents reported they would seek immediate medical care if they experienced stroke symptoms. Only 59.4% knew the emergency ambulance service telephone number.
CONCLUSIONIn a sample of Singaporean adults residing in a public housing estate, we found evidence of poor stroke literacy, highlighting the need for comprehensive population-based education efforts. There is a role for opportunistic education among those at higher risk of stroke.
Adult ; Aged ; Cross-Sectional Studies ; Female ; Health Literacy ; Humans ; Male ; Middle Aged ; Public Housing ; Risk Factors ; Singapore ; Socioeconomic Factors ; Stroke ; Surveys and Questionnaires ; Young Adult