類別:

龐貝氏症新生兒篩檢偽陽性事件對父母擔任親職之長期影響

2021-06-15T03:52:19Z

標題: 龐貝氏症新生兒篩檢偽陽性事件對父母擔任親職之長期影響; Long-term effect of false-positive Pompe disease newborn screening on parenting 作者: Hsiao-Ling Chen; 陳曉玲摘要: 新生兒篩檢是一種「以區域性人口族群」為背景的公共衛生政策，目的是降低某些先天代謝異常疾病的殘障率、疾病嚴重度或是死亡率。近年來，由於人類基因圖譜製作完成，衍生了許多新興科技、技術的發展與應用，也因此帶動了新生兒篩檢的發展，陸陸續續增加了新的疾病項目，龐貝氏症便是其中之一。本研究的目的是探討在某大醫院遺傳諮詢中心歷經龐貝氏症新生兒篩檢複檢及疾病確認事件後，篩檢結果為偽陽性之個案父母，其長期擔任親職時是否受到該複檢及疾病確認事件之影響。一共收集了研究組「有歷經龐貝氏症篩檢偽陽性的事件」之家長所填具的有效問卷19份，對照組「沒有歷經龐貝氏症篩檢偽陽性的事件」之家長所填具的有效問卷32份。研究的結果發現：研究組與對照組在「疾病不確定感」、「親職壓力」、「親職勝任感」三大主變項總分的比較中，在「疾病不確定感」(P=0.04)主變項達到顯著差異，對照組得分較高，表示研究組的疾病不確定感高於對照組。在「缺乏資訊」、「不明確」、「模糊不清」、「無法預測」、「親職愁苦」、「互動失調」、「困難兒童」、「效能」、「挫折」九項次級變項的比較中，在「缺乏資訊」(P=0.003)與「不明確」(P=0.006)次級變項達到顯著差異，且對照組得分高於研究組，顯示研究組在「缺乏資訊」與「不明確」向度上的疾病不確定感比對照組來的高。另外，將研究組分為「擔心」與「不擔心」篩檢結果2組，再作比較，結果發現「擔心」與「不擔心」在三大主變項的比較中，均未達顯著差異，但是在「親職壓力」主變項中有達到顯著差異的趨勢(P=0.055)；九項次級變項的比較中，則是在「互動失調」(P=0.008)次級變項中達到顯著差異，且在「親職愁苦」(P=0.057)次級變項中，有達到顯著差異的趨勢，且都是「不擔心」得分較高。此結果顯示「有歷經龐貝氏症篩檢偽陽性的事件」且在篩檢複檢與疾病確認過程中表達「擔心」的父母，在「互動失調」向度上的親職壓力，會比表達「不擔心」的父母高。將研究組與對照組之間在基本資料上達顯著差異的「家庭月收入」與「孩童年齡」項目，以Pearson’s Correlation分析所有的樣本，檢驗這2個項目與九項次級變項得分以及三大變項得分之間的的相關性；結果發現「孩童年齡」與九項次級變項之間皆未達顯著相關，只有與「缺乏資訊」(相關係數=-0.284)次級變項得分之間有達到顯著負相關的趨勢。「家庭月收入」則「困難兒童」(相關係數=+0.292)次級變項得分呈現顯著正相關，表示在本研究樣本中，「家庭月收入」越高，「困難兒童」次級變項的得分也會越高，也就是在「困難兒童」向度的親職壓力會越低。三大變項之間皆呈現顯著正相關，相關係數分列如下： *「疾病不確定感」與「親職壓力」之間相關係數為+0.456，「疾病不確定感」得分越高，「親職壓力」得分也越高，也就是不確定感越低，親職壓力越低。 *「疾病不確定感」與「親職勝任感」之間相關係數為+0.519，「疾病不確定感」得分越高，「親職勝任感」得分也越高，也就是不確定感越低，親職勝任感越高。 *「親職壓力」與「親職勝任感」之間相關係數為+0.729，「親職壓力」得分越高，「親職勝任感」得分也越高，也就是親職壓力越低，親職勝任感越高。顯示在本研究樣本中以「親職壓力」與「親職勝任感」之間的相關性最強，「疾病不確定感」與「親職勝任感」之間的相關性次之，「疾病不確定感」與「親職壓力」之間的相關性最弱。本研究中發現龐貝氏症新生兒篩檢偽陽性結果，長期來說並不會造成家長擔任親職時的親職壓力上升，但由於研究組與對照組在「疾病不確定感」主變項的比較中達到顯著差異，且本研究中也發現「疾病不確定感」、「親職壓力」、「親職勝任感」的得分有顯著正相關的關係；若是存有過高的不確定感，可能會造成親職勝任感降低與親職壓力變高。這些歷經篩檢偽陽性事件的家長們要面對的是孩子可能患有重大疾病的疑慮，以及後續是否進行基因檢測的選擇，因此醫療照護體系中的訊息傳達與關係建立可能還需要再特別加以關注，所以提供相關的遺傳諮詢服務是有必要的。; Newborn screening is a public health policy aims at reducing disability, severity of illness, or mortality of inborn errors of metabolism. After the success of the human genome project a few years ago, new science and technology allow more diseases to be screened at the newborn stage, like Pompe disease. The purpose of this study is to explore long-term effect of Pompe disease newborn screening on parenting. We conducted 19 questionnaires on the parents whose babies have experienced a false-positive Pompe disease screening (the study group). We also conducted 32 questionnaires on parents whose babies have a negative Pompe disease screening (the control group). We found that the total scores of 'Parenting Stress', and 'Parenting Sense of Competence' are not significantly different between the study group and the control group. There is significantly different between the study group and the control group in the total score of 'Uncertainty of Illness'. Among the nine sub-variables ('Lack of Information' , ' Lack of Clarity', 'Multiattributed Ambiguity', 'Unpredictability', 'Parental distress', ' Parent-Child dysfunctional interaction', 'Difficult Child', 'Efficiency', and 'Frustration'), we found that the scores of 'Lack of Information' and ' Lack of Clarity' sub-variable are higher in the control group than in the study group. This suggests that the study group is more uncertain about illness than the control group. This result is not related to whether the parents are 'worried' or 'not worried' about the screening result. But these parents, who expressed 'worried' about the screening result during false positive event, are perceiving more parenting stress in ' Parent-Child dysfunctional interaction' dimension than expressed 'not worried' about the screening result. One problem of the study is that the study group and the control group difference in both 'family income' and 'child age'. We analyze these two items in respective to either the three main variables or the nine sub-variables by Pearson's Correlation. We find that 'Family income' is positive correlated with 'Difficult Child'. Among the three main variables, each one is significantly positively correlated to the other two variables, respectively. In conclusion, a false positive Pompe disease newborn screening does not affect parenting significantly in 'Parenting Stress' and 'Parenting Sense of Competence', but the significantly different in 'Lack of Clarity' needs an attention. We need to be careful about delivering messages of newborn screening. Test instruction is very important, because the parents may face the fact having their children suffered from major illness. Service of genetic counseling is certainly needed.

高血脂症病人的分子基因學研究及相關脂蛋白之表現

2021-06-15T00:20:24Z

標題: 高血脂症病人的分子基因學研究及相關脂蛋白之表現; Molecular Analysis of the Taiwanese Patients with Hypertriglyceridemia and Expression of the Associated Apolipoprotein 作者: Yen-Hsin Chen; 陳彥心摘要: 由於現代人的飲食習慣與以往大為不同，加上忙碌的生活導致缺乏運動，以及一些抽菸、喝酒等生活習慣，使得代謝症候群的病人越來越多。其中常見的高血脂症(Hypertriglyceridemia，HTG)的發生不單單只受到後天環境因素的影響，先天遺傳基因也會造成三酸甘油脂(Triglycerides，TGs)升高；而TGs升高與心血管疾病(Coronary Artery Disease，CAD)及腦血管疾病(Cerebrovascular Disease)的相關性相當顯著(4,7,10,,17,24,31,55)。因此探究高血脂症的發生原因和先天性基因型的相互作用為我們重要之課題。首先，我們在代謝門診經專科醫師之診斷與協助，收集Controls (TGs<2.26mmol/L, n=285)，Moderate HTG (2.26≦TGs<5.65 mmol/L, n=216)，以及Severe HTG (TGs≧5.65mmol/L, n=169)三組之病患及對照組的檢體，並加以瞭解其基本資料和生活模式後，發現在年齡、BMI、腰圍、抽菸習慣和有無高血壓病史和HTG具有統計學的顯著相關(p-value<0.0001)，推論這些因素都可能會造成TGs升高，而引發高血脂症。接著，從臨床生化的檢驗數值發現，Severe HTG的病人在膽固醇(CHO)、TGs、飯前血糖(Glucose AC)和飯前胰島素(Insulin AC)上的表現比Moderate HTG和Control的病人較高(p-value<0.0001)，由此可見，血脂越高的病人在這些生化數值上的表現也都相對提高。探討後天環境的影響和臨床表徵之後，我們根據文獻選取與高血脂症發生有關的APOA5、APOE、APOC3三個候選基因(Candidate Gene)進行分析。若在APOA5基因上出現553G>T的polmorphism(APOA5:T(+))時，脂蛋白A5 (Apoliprotein A5)無法調節TGs的運輸，導致TGs升高；在APOC3基因的三端未轉錄區若有3238C>G的置換(S2基因型)時，脂蛋白C3 (Apolipoprotein C3)失去調控TGs代謝的功能，造成TGs升高；而在APOE基因上，倘若為APOE4的基因型態，在運送和代謝TGs、CHO的能力會變差，而使得血液中TGs的濃度升高。我們發現血脂表現越高，有APOA5之”T” allele及APOE4 allele的比例，在統計上明顯地增高(p-value<0.0001)。因為APOA5、APOE4和APOC3皆可能與高血脂症之發生有相關，所以利用Univariate and Multivariate Logistic Regression Aalysis去分析三個基因之間是否有交互作用，結果發現為APOA5:T(+)、APOC3:S2和APOE4基因型態時，最有機會造成高血脂症之發生，推論APOA5蛋白和APOE4同時存在，會造成TGs的調控不好，導致TGs升高。在脂蛋白元E4、C3和A5的表現上，發現脂蛋白C3的濃度與Lipid Profile(CHO、TGs、NHDL)的表現，具有統計意義(p-value<0.0001)，且脂蛋白元C3在TGs大於500 mg/dl的HTG表現會比血脂小於500 mg/dl的群體來的高。所以脂蛋白元C3的濃度與高血脂症有相關。在發現APOA5、APOC3和APOE4與高血脂症之關係後，我們可以建議高血脂症的病人進行基因型態之分析，幫助高血脂症病人之家屬改變飲食習慣和生活作息，以降低體內的血脂濃度，避免高血脂症之發生。對於病人，可以找到其高血脂症發生的原因，在併發症和情形尚未惡化之前，就接受治療，控制血脂，改善生活品質，並預防心血管疾病之發生。; The differences in eating habits and less exercise influence people’s health condition. The increasing people suffered from metabolic syndrome including Hypertriglyceridemia (HTG) which results from genetic and environmental factors. Some researches say that the elevated triglycerides (TGs) are related to coronary artery disease (CAD) and cerebrovascular disease. We supposed to do is consider what caused HTG and clarify the interaction of genotypes. We collect three groups of Controls (TGs<2.26mmol/L, n=285), Moderate HTG (2.26≦TGs<5.65 mmol/L, n=216), and Severe HTG (TGs≧5.65mmol/L, n=169) from Dr. Su’s metabolic clinic in National Taiwan University Hospital. Get information of their age, BMI, waist, smoking habit and the history of hypertension associated with HTG patients, and identify their significance in statistics (p-value<0.0001). And then consider the biochemical analysis, we find the expressions of cholesterol (CHO), TGs, Glucose AC, Insulin AC in severe HTG are higher than moderate HTG and Controls (p-valus<0.0001). According to previous papers, we pick up three candidate gene (APOA5, APOE and APOC3) to do genetic analysis. If we find the 553G>T polymorphism in APOA5 gene, apolipoprotin A5 cannot modulate the transport of TGs and cause TGs increase. The replacement of C to G occurred in 3’ untranslated region at 3238 site of APOC3 gene, which make apolipoprotein C3 fail to regulate TGs metabolism. The consequence is TGs elevated. And if the genotype is APOE4, the function of TGs transport and metabolism will not work well and the concentration of TGs will be high. There are statistic significances (p-value<0.0001) in high ratio of T allele in APOA5 and APOE4 allele. Consequently, APOA5, APOE4 and APOC3 may be related to HTG. Therefore we use univariate and multivariate logistic regression analysis to analyze the interaction between these three genes. We postulate the co-existence of apoliprotein A5 and apolipoprotein E4 influence the function of TGs regulation and then TGs increase. Otherwise, we also observe the protein level of apolipoprotein E4, C3 and A5. Then, we find that the concentration of apolipoprotein C3 is associated with lipid profile (p-value<0.0001). As a result, we assume that the level of apolipoprotein C3 is associated with TGs level in HTG patients. Clarify APOA5, APOE4, and APOC3 are associated with HTG. What we should do is suggest the patients with HTG do genetic analysis to confirm genotype, and then help them change their life style to control TGs level and avoid coronary artery disease.

高肌酸激酶血症新生兒篩檢技術之開發

2021-06-16T05:09:45Z

標題: 高肌酸激酶血症新生兒篩檢技術之開發; Development of a newborn screening technique for hyper-creatine kinase-emia 作者: Li-Lan Liu; 劉麗蘭摘要: 背景肌酸激酶(Creatine kinase, CK)是參與體內能量代謝的一種酵素，也是一種可逆磷酸化的酶，CK可形成高能量的三磷酸腺苷ATP，CK亦能供給肌肉能量的來源。在臨床上，測定血清CK檢查指的是血清中total CK的活性，由於在骨骼肌，心臟肌肉，和腦組織中含有豐富的激酸激酶。因此常用於肌肉相關疾病的檢查，如急性心肌梗塞，當這些部位受傷，血清中的CK會上升。在龐貝氏症、肢帶型肌肉失養症、心肌梗塞患者及肌營養不良症(如裘馨氏肌營養不良症及貝克型肌肉營養不良症)的血清中CK活性均呈現輕度至中度升高。而和CK的相關研究也顯示在裘馨氏肌肉失養症的男性患者CK活性會顯著升高，女性患者的CK活性則會輕微升高。方法本研究方法是利用血片檢體，做大規模的篩檢，以免疫螢光法檢測儀檢測血片中肌酸激酶的活性。結果本實驗總共分析有8,083個新生兒血片，其中男性有4,178個(51.7 %)，女性有3,905個(48.3%)，男女比例為1.07，在8,083個檢體中，CK值在1500 Flu/min以上共有10個檢體，佔0.12%，其中男性檢體有5人，女性檢體有5人，CK值在1,200至1,500 Flu/min有12個檢體，佔0.15%，男性檢體有7人，女性檢體有5人，CK值在1,000至1,200 Flu/min有23個檢體，佔0.28%，男性檢體有19人，女性檢體有4人。結論 CK是一種來自肌肉的酵素，因此在肌肉受傷時，會引起CK的上升，本實驗方法是利用新生兒篩檢血片進行大量、快速，且有效的CK活性篩檢，找出高CK疑陽性個案。我們的研究結果，CK Flu/min >= 1,000共計有45個檢體為高疑陽性個案。在CK與其他因子的關連性研究的結果顯示，在CK與性別的關連性上，其在統計上沒有顯著差異(P > 0.05)。在CK與出生體重的關連性上，以體重小於或大於等於2,200 g做分組研究，在統計上具有顯著差異(P < 0.05)。在CK與妊娠週數的關連性上，以妊娠週數小於或大於等於37週做分組研究，在統計上具有顯著差異(P < 0.05)。在CK與CAH的關連性上，以妊娠週數大於等於37週做分組研究，統計上具有顯著差異(P < 0.05)。CK活性的確有可能因生產過程的擠壓而略為升高，但應該不會影響高疑陽性個案的判斷。在CK與性G6PD的關連性上，結果顯示G6PD缺乏症患者的CK數值並不會有偏低之情形發生。G6PD缺乏症應該不會影響CK反應下降，造成偽陰性的判斷。; Background Creatine kinase, a reversible phosphorylation enzyme, is one of the enzymes involved in energy metabolism. It can form a high energy triphosphate ATP and provide the muscle energy. Clinically, serum CK examination refers to the total CK activity in serum. Due to the significant amount of kinase in skeletal and heart muscle, and brain.CK is commonly used as an indicator to examine the muscle-related disorders. When injuries of muscle, brain and heart occur, the CK level is usually elevated. In most patients with Pompe disease, limb-girdle muscular dystrophy, myocardial infarction, and muscular dystrophy (such as Duchenne muscular dystrophy and Becker muscular dystrophy), CK activity also showed mild to moderate elevations. In the cases of Duchenne muscular dystrophy, the serum CK activity is significantly increased in male patients but only slightly elevated in female patients. Method Our research method use the dried blood spot samples for mass screening to detect creatine kinase activity by fluorescence detector . Results In this study, a total of 8,083 newborns dried blood spots were analyzed. Among them, there were 4,178 male (51.7%) and 3,905 female (48.3%), and the male to female ratio was 1.07. The CK Flu / min values greater than 1,500 or more was detected in 10 cases (0.12%) of which male vs. females were equal. The CK Flu / min values of 1,200-1,500 was found in 12 samples (0.15%)indicated 7 male and 5 female subjects. The CK Flu / min values in 1,000-1,200 was detected in 23 samples (0.28%) 19 male and 4 female subjects. Conclusions Clinically, CK is commonly used as an indicator to examine the muscle-related disorders. In this study, we perform a rapid and effective high-through put screening using the newborn screening dried blood spots. From the results, there are 45 cases with higher CK activity (>=1,000 CK Flu/min) in the DBS. In the study of CK and other parameters, we find no relation between CK and Gender, but there are relationship between CK & Birth weight and CK & Gestational age. From the CK and CAH study, birth Stress may certainly make some raise in the CK level, but it may not influence the judgement of hyper-creatine kinase-emia. Although we use the reagents with G6PD enzyme, the G6PD enzyme deficiency in the DBS may not decrease the CK reaction obviously.

高滲透壓甘油反應磷酸化激酶調控鳥糞嘌呤核苷酸交換因子以活化第一腺嘌呤核苷二磷酸核醣化相似因子之探討

2021-06-17T02:30:07Z

標題: 高滲透壓甘油反應磷酸化激酶調控鳥糞嘌呤核苷酸交換因子以活化第一腺嘌呤核苷二磷酸核醣化相似因子之探討; Characterization of Hog1p protein kinase in regulating Syt1p-mediated Arl1p activation 作者: Yi-Hsun Wang; 王奕勛摘要: 腺嘌呤核苷二磷酸核醣化相似因子 (ARF-like protein 1, ARL1) 屬於腺嘌呤核苷二磷酸核醣化因子家族的成員之一 (ADP-ribosylation factors, ARFs)。在真核生物中，這類蛋白被報導參與許多重要的細胞功能，包含調控囊泡運輸、影響細胞骨架的組裝以及維持細胞內的離子恆定等等，但是作用機制的細節仍不甚清楚。在酵母菌的研究當中，已知鳥糞嘌呤核苷酸交換因子 (Guanine nucleotide exchange factor) Syt1p 會催化 Arl1p 的活性並使其前往反式高基氏體網路 (trans-Golgi network)。活化態的 Arl1p 將吸引高基氏體蛋白 (Imh1p) 至反式高基氏體網路並作用於下游的囊泡運輸。在我們先前的發表成果中，我們發現內質網壓力 (ER stress) 的誘發將促使內質網上的磷酸化激酶 Ire1p 活性上升，Ire1p將透過 Syt1p 的第416號絲氨酸磷酸化轉譯後修飾進一步大量活化 Arl1p。此外，我們也證實這條路徑的啟動有助於細胞抵擋內質網壓力。而在本篇研究中，我們發現在細胞經歷內質網壓力的過程中， Hog1p 是另一個調控 Syt1p的磷酸化激酶。不同於 Ire1p 的是，Hog1p 作用於 Syt1p 的第297號絲氨酸磷酸化轉譯後修飾以活化 Arl1p。同時，我們亦發現不受壓力激活的 Hog1p 也可以調控 Arl1p 的活化，說明在壓力環境以及正常生理狀態下，Hog1p 皆扮演活化 Arl1p 的重要角色。總結本篇除了再次說明 Syt1p 的磷酸化轉譯後修飾對於 Arl1p 活化的重要性，我們亦提出 Hog1p 在正常生理狀態中所執行的新功能。; ARF-like (ARL) proteins play important roles in regulating vesicular trafficking at the Golgi compartments, modulating cytoskeleton dynamics and maintaining ion homeostasis. In Saccharomyces cerevisiae, guanine-nucleotide exchange factor (GEF) Syt1p facilitates Arl1p activation to recruit golgin protein Imh1p to trans-Golgi network (TGN). Our previous findings have elucidated the induction of unfolded protein response (UPR) highly promotes activation of Arl1p and that ER-resident kinase Ire1p is responsible for the up-regulation through transcription alteration (Proc. Natl. Acad. Sci. U S A. 113:E1683-E1690). Moreover, the SYT1-ARL1-IMH1 signaling is required for ER stress resistance. In this study, we identified MAP kinase Hog1p as a new regulator of the ARL1 pathway for UPR. We found that MAP kinase controls Arl1p activation and non-stressed Hog1p mutant can rescue Imh1p Golgi localization in MAP kinase-deleted cells. We further showed that Hog1p might directly phosphorylate Syt1p at serine 297 to promote the activation of Arl1p, which performs a distinct route from Ire1p-mediated Syt1p phosphorylation at serine 416. Collectively, we demonstrate that Syt1p phosphorylation is regulated by multi-regulators, which is important for the activation and function of Arl1p under stress as well as normal growth conditions.