Y型聚乙二醇干扰素琢-2b注射液治疗HCV基因2/3型慢性丙型肝炎患者疗效和安全性的多中心随机对照试验研究

目的以标准剂量的聚乙二醇干扰素(Peg IFN)α-2a联合利巴韦林作为阳性对照,评价新型试验药物Y型Peg IFNα-2b注射液联合利巴韦林治疗2型/3型慢性丙型肝炎(CHC)患者的疗效和安全性。方法采用多中心、随机开放、阳性药对照的Ⅲ期临床试验,筛选符合要求的2型/3型CHC患者,按照2:1的比例随机分配到Y型Peg IFNα-2b组和Peg IFNα-2a组,同时口服利巴韦林,疗程24 w,停药随访24 w。采用Abbott Real Time HCV Genotype II检测HCV基因型,采用Cobas Taq Man实时定量PCR法检测血清HCV RNA水平。详细记录不良事件。主要疗效指标为持续病毒学应答(SVR),并进行非劣效检验。结果本试验实际入组2型/3型CHC患者255例,实际治疗241例。全分析集(FAS)数据显示,158例试验组和83例对照组患者SVR分别为85.4%(95%CI 79.94%~90.94%)和79.5%(95%CI 70.84%~88.20%,P=0.2402);对符合方案分析集(PPS)人群分析显示,试验组和对照组患者SVR分别为87.9%(95%CI 82.45%~93.27%)和85.9%(95%CI 77.82%~94.01%,P=0.7060),率差的95%可置信区间均符合非劣效标准;对PPS人群分析显示,85.8%受试者获得了早期病毒学应答(RVR),RVR的阳性预测值为90.1%;试验组和对照组不良事件发生率相似,分别为95.6%和95.2%,严重不良事件发生率分别为3.8%和3.6%。结论应用Peg IFNα联合利巴韦林治疗2型/3型CHC患者,新型试验药物Y型Peg IFNα-2b具有与对照药物Peg IFNα-2a相似的疗效和安全性。国家科技部“十二五”重大专项(编号:2012ZX10002-003)；“重大新药创制”十二五科技重大专项(编号:2012ZX09303019)

Docosahexaenoic acid enhances hepatic serum amyloid A expression via protein kinase A-dependent mechanism

血清澱粉狀蛋白A ( serum amyloid A, SAA) 能減少人類肝細胞和脂肪細胞中脂肪堆積並且SAA在這些細胞的表現會因二十二碳六烯酸 ( docosahexaenoic acid, DHA) 處理而增加。藉由了解DHA是否媒介SAA以及以何種機制來減少脂肪堆積將有助於發展對抗肥胖和脂肪肝的新策略。本試驗先對數種常用的肝癌細胞株進行篩選，結果顯示DHA處理後SAA1在SK-HEP-1表現會增加，在PLC/PRF/5和Hep3B會減少。因為先前發現富含DHA的飼糧會增加豬隻肝臟SAA的表現，所以SK-HEP-1便被用於下列的實驗。SK-HEP-1細胞處理100及200 μM的DHA會增加SAA1和其上游調控因子CCAAT/enhancer binding protein beta ( C/EBPβ ) mRNA的表現。利用啟動子刪除分析法可得知在SAA1啟動子上為於-242 and -102的片段對DHA所調控的SAA1表現很重要，而此片段包含有C/EBPβ結合點。當突變C/EBPβ結合點上的DNA序列使其失去與C/EBPβ結合的能力後，因DHA處理所增加的promoter activity就會受到抑制，這顯示出此C/EBPβ結合點參與DHA對SAA1的調控。本試驗亦發現因DHA處理所增加SAA和C/EBPβ蛋白質表現會因PKA抑制劑 (H89) 的添加而受到抑制，此可顯示C/EBPβ是經由PKA所媒介而參與DHA對SAA1的調控。除此之外對於DHA減少脂肪細胞和肝細胞中脂肪堆積的機制也進行相關研究。DHA處理會增加脂肪細胞甘油的釋出。為了解DHA對脂肪肝的效應，SK-HEP-1在5 mM oleic acid處理2天後，細胞內脂質堆積明顯增加。以100 μM DHA處理這些脂肪肝細胞後，其細胞內的三酸甘油脂會減少。DHA對抑制人類肝細胞和脂肪細胞中脂肪堆積的效應亦會因H89的添加而消失，顯示DHA對脂肪堆積的效應也是由PKA所媒介。綜言之，由於DHA對SAA1的正向調控和脂肪堆積的負向調控皆經由PKA這條路徑，所以可以推論DHA可能是藉由SAA1減少脂肪堆積。這些發現對於了解DHA所調控的脂肪代謝機制提供了新線索，此將有助於DHA在改善人類健康上的應用。Serum amyloid A (SAA) reduces fat deposition in adipocytes and hepatocytes. Human SAA1 mRNA is increased by docosahexaenoic acid (DHA) treatment in human cells. These studies asked whether DHA decreases fat deposition through SAA1 and explored the corresponding mechanisms. We demonstrated that DHA increased human SAA1 and C/EBPβ mRNA expression in human hepatoma cells, SK-HEP-1. Utilizing promoter deletion assay, we found that a C/EBPβ binding site in the SAA1 promoter region between -242 and -102 bp is critical for DHA-mediated SAA1 expression. When we mutated the putative C/EBPβ binding site, the DHA-induced SAA1 promoter activity was suppressed, suggesting that this binding sequence was very important for the DHA regulated transcription activation. The addition of the protein kinase A inhibitor, H89, negated the DHA-induced increase in C/EBPβ protein expression. The up-regulation of SAA1 mRNA and protein by DHA was inhibited by H89 treatment, as well. We further demonstrated that DHA increased protein kinase A (PKA) activities. These data suggest that C/EBPβ is involved in the DHA-regulated increase in SAA1 expression via PKA-dependent mechanisms. Furthermore, the suppressive effect of DHA on triacylglycerol accumulation was abolished by H89 treatment in hepatocytes and adipocytes indicating that DHA reduces lipid accumulation via PKA as well. The phenomena of increased SAA1 expression coupled with reduced fat accumulation mediated by DHA via PKA suggest that SAA1 is involved in DHA-induced triacylglycerol breakdown. These findings provide new insights into the complicated regulatory network in DHA-mediated lipid metabolism and may be useful in developing new therapeutic approaches to reduce body fat deposition and fatty liver.Contentsage試委員會審定書…………………………………………………………………… i cknowledgement…………………………………………………………………….. iihinese Abstract……………………………………………………………………... iiinglish Abstract………………………………………………………………………... ivontents……………………………………………………………………………....... viist of Figures………………………………………………………………………... viii ist of Tables………………………………………………………………………….. ixhapter 1 Literature Review………………………………………………………… 1.1 Introduction.2 n-3 PUFA on lipid metabolism.3 Inflammation mediators involved in n-3 PUFA-regulated lipid metabolism.4 Inflammation mechanisms of n-3 PUFA.5 Summaryhapter 2 Special aims………………………………………………………….. 23hapter 3 Materials and Methods……………………………………………….. 24 3.1 Hepatoma cells and culture media.2 Human adipose tissue samples and adipocytes isolation and differentiation .3 Real-time PCR.4 Western blotting and ELISA.5 Plasmid construction, Transient transfection and Dual Luciferase Assay.6 PKA activity.7 Staining.8 Triacylglycerol measurements.9 Statistical analysishapter 4 Results……………………………………………………………………. 34 4.1 Effect of DHA on SAA1 expression 4.2 Effect of DHA on SAA1 upstream regulatory transcription factor 4.3 Role of C/EBPβ in DHA-mediated SAA1 promoter activity 4.4 Role of PKA in DHA-mediated SAA1 and C/EBPβ expression 4.5 Effect of DHA on PKA activities 4.6 In vitro fatty hepatocyte model setup 4.7 Effect of DHA on triacylglycerol breakdownhapter 5 Discussion………………………………………………………………… 48hapter 6 References………………………………………………………………… 5

Study on Potential and Affected Area Analysis-An Example of Nantou County

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Experimental study on indoor thermal environment of active solar heating in new rural residential construction

Solving the problem of the new rural residential heating in winter is a technical barrier of green energy-saving building.This paper was studied the experimental comparison of two same toward the size of the new rural residential building indoor air temp

Evaluating the urate-lowering effects of different microbial fermented extracts in hyperuricemic models accompanied with a safety study

[[abstract]]Uric acid (UA) is an end product of purine metabolism by the enzyme xanthine oxidase (XOD). Hyperuricemia is characterized by the accumulation of serum UA and is an important risk factor for gout and many chronic disorders. XOD inhibitors or uricase (catalyzes UA to the more soluble end product) can prevent these chronic diseases. However, currently available hypouricemic agents induce severe side effects. Therefore, we developed new microbial fermented extracts (MFEs) with substantial XOD inhibition activity from Lactobacillus (MFE-21) and Acetobacter (MFE-25), and MFE-120 with high uricase activity from Aspergillus. The urate-lowering effects and safety of these MFEs were evaluated. Our results showed that MFE-25 exerts superior urate-lowering effects in the therapeutic model. In the preventive model, both MFE-120 and MFE-25 significantly reduced UA. The results of the safety study showed that no organ toxicity and no treatment-related adverse effects were observed in mice treated with high doses of MFEs. Taken together, the results showed the effectiveness of MFEs in reducing hyperuricemia without systemic toxicity in mice at high doses, suggesting that they are safe for use in the treatment and prevention of hyperuricemia