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Effect of intestinal ischemia/reperfusion injury on leptin and orexin-A levels

LIN Ji, YAN Guangtao, HAO Xiuhua, ZHANG Kai, GAO Xiaoning, LIAO Jie

《医学前沿(英文)》 2007年 第1卷 第1期   页码 87-92 doi: 10.1007/s11684-007-0017-3

摘要: The aim of this paper is to explore the effect of intestinal ischemia/reperfusion (I/R) injury on leptin and orexin-A levels in peripheral blood and central secretory tissues, and to examine the roles of leptin and orexin-A in acute inflammatory responses. An intestinal I/R injury model of rats was made; the rats were grouped according to the time of after 60 min ischemia. Radioimmunoassay was employed to detect the levels of leptin in serum and adipose tissue and orexin-A levels in plasma and hypothalamus. Reverse transcriptase-polymerase chain reaction was used to detect mRNA expressions of adipose leptin and hypothalamus orexin-A. Compared with the levels before the injury, serum leptin in 60 min ischemia/30 min reperfusion (I60´R30´) group decreased and that of I60´R360´ group increased. Compared with sham-operation group (sham group) after injury, serum leptin level of I60´R360´ group increased, adipose leptin levels of I60´R30´ and I60´R90´ decreased, and adipose leptin in I60´R360´ group increased. After the injury, adipose leptin mRNA expressions of I60´R30´, I60´R240´ and I60´R360´ increased, whereas that of I60´R150´ group decreased as compared with the sham group. There was no significant difference in the protein levels of orexin-A, either between plasma and hypothalamus or between pre- and post-I/R injury. Compared with sham group, hypothalamus orexin-A mRNA expressions of I60´R30´ and I60´R90´ decreased gradually after the injury, with that of I60´R150´ group reaching the lowest, and those of I60´R240´ and I60´R360´ recovering gradually, although they were still significantly lower than that of sham group. Leptin and orexin-A respond to intestinal I/R injury in a time-dependent manner, with leptin responding more quickly than orexin-A does, and both of them may contribute to the metabolic disorders in acute inflammation.

关键词: significant difference     intestinal I/R     transcriptase-polymerase     metabolic     central secretory    

Abnormal expression of c-Myc in human bronchial epithelial cells malignantly transformed by anti-BPDE

FU Juan, JIANG Yiguo, CHEN Xuemin

《医学前沿(英文)》 2008年 第2卷 第4期   页码 380-385 doi: 10.1007/s11684-008-0073-3

摘要: Anti-benzo[a]pyrene-7,8-diol-9,10-epoxide (anti-BPDE) is a metabolite of benzo[a]pyrene (B[a]P) and acts as a potent mutagen in mammalian systems. However, molecular mechanisms related to anti-BPDE-induced carcinogenesis are poorly understood. Here, we investigated the expression of proto-oncogene c- in human bronchial epithelial cells (16HBE-T) transformed by exposure to anti-BPDE. The levels of mRNA and protein of c-Myc were examined in the 16HBE-T and vehicle-treated control cells (16HBE-N) by using different methods respectively, including reverse transcriptase-polymerase chain reaction (RT-PCR), quantitative real-time PCR (Q-PCR), western blot and immunocytochemical methods. The level of c- mRNA appeared to be significantly increased in 16HBE-T, as compared with those of the 16HBE-N. Likewise, the expression of c-Myc protein was significantly enhanced as compared with those of the control cells. Moreover, the localization of c-Myc protein shows mainly nuclear staining in 16HBE-T. In conclusion, the abnormal expression of c-Myc was present in anti-BPDE malignantly transformed 16HBE cells, which may be involved in the carcinogenesis molecular mechanism of anti-BPDE.

关键词: transcriptase-polymerase     vehicle-treated     understood     molecular mechanism     different    

Effects of 3-aminobenzamide on poly (ADP-ribose) polymerase expression, apoptosis and cell cycle progression

DU Xiang, ZHAO Hongguang, GUO Wei, GONG Shouliang, WANG Wen

《医学前沿(英文)》 2008年 第2卷 第2期   页码 204-206 doi: 10.1007/s11684-008-0039-5

摘要: The aim of this paper is to study the changes of apoptosis and cell cycle progression in HeLa cells after the poly (ADP-ribose) polymerase (PARP) was inhibited by its inhibitor 3-aminobenzamide (3-AB) and the mechanisms of PARP action on HeLa cells damaged by irradiation. Flow cytometry (FCM) was used to examine the PARP expression and the percentage of apoptotic cells and cell cycle progression. The percentage of HeLa cells with positive expression of PARP protein 2, 4, 8 and 12 h after administrated with 3-AB was significantly lower than that of the control ( < 0.01). The percentages of apoptotic cells in the 3-AB plus irradiation group at the time points of 2, 8, 12 and 24 h after 2 Gy irradiation were higher than that in the irradiation group ( < 0.01 or < 0.05) and the percentage of G cells decreased significantly ( < 0.01 or < 0.05). It indicates that 3-AB can rapidly inhibit PARP expression of HeLa cells, promote cell apoptosis and block G arrest induced by irradiation.

关键词: control     irradiation     apoptotic     progression     3-AB    

Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

《环境科学与工程前沿(英文)》 2011年 第5卷 第1期   页码 21-27 doi: 10.1007/s11783-011-0303-9

摘要: New high-throughput technologies continue to emerge for studying complex microbial communities. In particular, massively parallel pyrosequencing enables very high numbers of sequences, providing a more complete view of community structures and a more accurate inference of the functions than has been possible just a few years ago. In parallel, quantitative real-time polymerase chain reaction (QPCR) allows quantitative monitoring of specific community members over time, space, or different environmental conditions. In this review, the principles of these two methods and their complementary applications in studying microbial ecology in bioenvironmental systems are discussed. The parallel sequencing of amplicon libraries and using barcodes to differentiate multiple samples in a pyrosequencing run are explained. The best procedures and chemistries for QPCR amplifications are also described and advantages of applying automation to increase accuracy are addressed. Three examples in which pyrosequencing and QPCR were used together to define and quantify members of microbial communities are provided: in the human large intestine, in a methanogenic digester whose sludge was made more bioavailable by a high-voltage pretreatment, and on the biofilm anode of a microbial electrolytic cell. The key findings in these systems and how both methods were used in concert to achieve those findings are highlighted.

关键词: polymerase chain reaction (PCR)     microbial communities     pyrosequencing     gut     microbial fuel cell     sludge    

Advances in newly developing therapy for chronic hepatitis C virus infection

null

《医学前沿(英文)》 2014年 第8卷 第2期   页码 166-174 doi: 10.1007/s11684-014-0334-2

摘要:

Chronic hepatitis C virus (HCV) infection afflicts a reported 170 million people worldwide and is often complicated by cirrhosis and hepatocellular carcinoma. Morbidity and mortality are decreased with the successful treatment of chronic HCV infection. Increased understanding of the HCV has allowed further development of new direct-acting antiviral (DAA) agents against the HCV and has also allowed the development of IFN-free oral treatment regimens. In late 2013 the first nucleotide polymerase inhibitor regimen with RBV alone for genotypes 2/3 and in combination with a 12-week regimen of PEG-IFN+RBV for genotypes 1, 4 was approved for use in the US. A number of promising new DAA regimens which are IFN-free are in phase 3 development and the first will likely be approved for use in the US in 2014. The currently approved regimens are discussed in detail and currently available data on future regimens are reviewed herein.

关键词: direct-acting antiviral (DAA)     nucleotide polymerase inhibitors     protease inhibitors    

低成本PCR微反应器阵列的温度测量与控制技术

李现明,张玉林,李建田

《中国工程科学》 2007年 第9卷 第7期   页码 76-81

摘要:

设计了一种用于聚合酶链式反应的低成本微反应器,其内部无微加热器、微传感器。针对这种低成本的微反应器,提出了一种温度测量与控制方案,其特点是将各反应器温度的集中控制与分散控制有机结合,能够同时完成上百只微反应器快速、准确的温度循环。重点阐述了温度传感器负荷效应的提取与补偿、动态误差补偿、微反应器温度间接测量误差评定、微反应器温度串级控制策略。

关键词: 微反应器     测量     控制     聚合酶链式反应    

Using RAPD in genotyping and transmission detection

ZHANG Tiejun, ZHOU Xiaoming, ZHANG Tao, YU Shunzhang, JIANG Qingwu, CHEN Yue

《医学前沿(英文)》 2008年 第2卷 第3期   页码 269-275 doi: 10.1007/s11684-008-0051-9

摘要: The aim of this paper is to develop an applicable Random Amplified Polymorphic DNAs (RAPD) method for genotyping strain, and discuss the possibility of using the RAPD method to trace strain transmission route. Four different pretreatment methods were used on the genomic DNA component, and the best adaptive extract method was selected for RAPD. Different RAPD primers sequence was used for amplification and their differentiating capabilities for strains were compared. Applicable RAPD primer was selected for genotyping and then applied into transmission detection. The results show that the so called cetyltrimethylammonium bromide (CTAB) method for extracting genomic DNA could give integrated genomic DNA and give out relatively better RAPD fingerprint maps, subsequently, using selected RAPD primer could give out a group of amplification polymerase chain reaction bands. The fingerprint maps from different strains were distinctive. Some main segments were common to all the strains tested. Some segments were different among the strains. According to the fingerprint maps and similarity index of different isolates, isolates from a pair of sex-partners were very similar. Based on these findings, the best extracting method and suitable RAPD primer were chosen. The RAPD fingerprint maps could type effectively and could be used as an additional approach in molecular epidemiology for tracing infection sources.

关键词: amplification polymerase     CTAB     sequence     cetyltrimethylammonium     RAPD    

Expression and clinical implication of PRL-1 and PRL-3 in transitional cell carcinoma of bladder

Bin HAO, Changwei LIU, Huixiang LI

《医学前沿(英文)》 2009年 第3卷 第2期   页码 197-203 doi: 10.1007/s11684-009-0036-3

摘要: The mRNA and protein expression of phosphatase of regenerating liver 1 (PRL-1) and phosphatase of regenerating liver 3 (PRL-3) in transitional cell carcinoma of bladder (BTCC) and normal epithelia of bladder was investigated, and the relationship between the BTCC and pathological changes was clarified. The expression of PRL-1 and PRL-3 mRNA was detected by using reverse transcription polymerase chain reaction (RT-PCR) in 30 cases of BTCC and 10 cases of normal bladder, and the expression of PRL-1 and PRL-3 protein was checked by using immunohistochemistry in 30 cases of BTCC and 15 cases of normal bladder. The expression levels of PRL-1 and PRL-3 mRNA and protein were higher in BTCC than those in normal bladder epithelia ( <0.05). The increased expression of PRL-1 and PRL-3 mRNA and protein was detectable in deep invasion and metastasis of BTCC ( <0.05). There was no correlation between the expression of PRL-1 and PRL-3 and gender, age or recurrence of BTCC (all >0.05). A significantly positive correlation was found between PRL-1 and PRL-3 in BTCC ( <0.05). PRL-1 and PRL-3 are expressed consistently and may contribute to the growth, differentiation, invasion and metastasis of BTCC.

关键词: transitional cell carcinoma of bladder     phosphatase of regenerating liver 1     phosphatase of regenerating liver 3     reverse transcription polymerase chain reaction     immunohistochemistry    

Seasonal microbial community shift in a saline sewage treatment plant

Qingmei YAN, Xuxiang ZHANG, Tong ZHANG, Herbert H P FANG

《环境科学与工程前沿(英文)》 2011年 第5卷 第1期   页码 40-47 doi: 10.1007/s11783-011-0304-8

摘要: Activated sludge was monthly sampled from a saline sewage treatment plant of Hong Kong (China) during June 2007 to May 2008 to analyze the microbial community shift along with environmental variations using denaturing gradient gel electrophoresis of polymerase chain reaction amplified 16S rDNA fragments. Environmental changes resulted into a seasonal microbial community shift characterized by alterations in species number and abundance in the sewage treatment plant. Correspondence analysis and cluster analysis on community structure profile showed that the 12 monthly samples fell into four groups, which is in accordance with season changing in Hong Kong. Canonical correspondence analysis revealed that and posed more significant effects on community structure than total phosphorus and total nitrogen, respectively. Compared with sludge retention time, influent total phosphorus had an inverse effect on the community structure shift, and chemical oxygen demand and showed a similar effects. Results of this study may contribute to the development of new knowledge involving the microbial community shift in sewage treatment plants.

关键词: sewage treatment plants     polymerase chain reaction (PCR)     denaturing gradient gel electrophoresis (DGGE)     canonical correspondence analysis     correspondence analysis    

Interaction and independence on methane oxidation of landfill cover soil among three impact factors: water, oxygen and ammonium

Pinjing HE, Na YANG, Wenjuan FANG, Fan Lü, Liming SHAO

《环境科学与工程前沿(英文)》 2011年 第5卷 第2期   页码 175-185 doi: 10.1007/s11783-011-0320-8

摘要: To understand the influence patterns and interactions of three important environmental factors, i.e. soil water content, oxygen concentration, and ammonium addition, on methane oxidation, the soils from landfill cover layers were incubated under full factorial parameter settings. In addition to the methane oxidation rate, the quantities and community structures of methanotrophs were analyzed to determine the methane oxidation capacity of the soils. Canonical correspondence analysis was utilized to distinguish the important impact factors. Water content was found to be the most important factor influencing the methane oxidation rate and Type II methanotrophs, and the optimum value was 15% (w/w), which induced methane oxidation rates 10- and 6- times greater than those observed at 5% (w/w) and 20% (w/w), respectively. Ambient oxygen conditions were more suitable for methane oxidation than 3% oxygen. The addition of of ammonium induced different effects on methane oxidation capacity when conducted at low or high water content. With regard to the methanotrophs, Type II was sensitive to the changes of water content, while Type I was influenced by oxygen content. Furthermore, the methanotrophic acidophile, , was detected in soils with a pH of 4.9, which extended their known living environments.

关键词: quantitative polymerase chain reaction (PCR)     denaturing gradient gel electrophoresis (DGGE)     principal component analysis (PCA)     canonical correspondence analysis (CCA)    

Cultivation of aerobic granular sludge in a conventional, continuous flow, completely mixed activated sludge system

Xi CHEN,Linjiang YUAN,Wenjuan LU,Yuyou LI,Pei LIU,Kun NIE

《环境科学与工程前沿(英文)》 2015年 第9卷 第2期   页码 324-333 doi: 10.1007/s11783-014-0627-3

摘要: Aerobic granules were formed in a conventional, continuous flow, completely mixed activated sludge system (CMAS). The reactor was inoculated with seed sludge containing few filaments and fed with synthetic municipal wastewater. The settling time of the sludge and the average dissolved oxygen (DO) of the reactor were 2 h and 4.2 mg·L , respectively. The reactor was agitated by a stirrer, with a speed of 250 r·min , to ensure good mixing.The granular sludge had good settleability, and the sludge volume index (SVI) was between 50 and 90 mL·g . The laser particle analyzer showed the diameter of the granules to be between 0.18 and 1.25 mm. A scanning electron microscope (SEM) investigation revealed the predominance of sphere-like and rod-like bacteria, and only few filaments grew in the granules. The microbial community structure of the granules was also analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Sequencing analysis indicated the dominant species were α, β, and γ- , , and . The data from the study suggested that aerobic granules could form, if provided with sufficient number of filaments and high shear force. It was also observed that a high height-to-diameter ratio of the reactor and short settling time were not essential for the formation of aerobic granular sludge.

关键词: aerobic granular sludge     completely mixed activated sludge system (CMAS)     continuous flow     shear force     filamentous bacteria     polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE)    

Design and use of group-specific primers and probes for real-time quantitative PCR

Juntaek LIM, Seung Gu SHIN, Seungyong LEE, Seokhwan HWANG

《环境科学与工程前沿(英文)》 2011年 第5卷 第1期   页码 28-39 doi: 10.1007/s11783-011-0302-x

摘要: Real-time quantitative polymerase chain reaction (qPCR) has gained popularity as a technique to detect and quantify a specific group of target microorganisms from various environmental samples including soil, water, sediments, and sludge. Although qPCR is a very useful technique for nucleic acid quantification, accurately quantifying the target microbial group strongly depends on the quality of the primer and probe used. Many aspects of conducting qPCR assays have become increasingly routine and automated; however, one of the most important aspects, designing and selecting primer and probe sets, is often a somewhat arcane process. In many cases, failed or non-specific amplification can be attributed to improperly designed primer-probe sets. This paper is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR assays. We demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic nutrient removal processes. qPCR assays using group-specific primers and probes designed with this method, have been used to successfully quantify 16S ribosomal Ribonucleic Acid (16S rRNA) gene copy numbers from target methanogenic and ammonia- oxidizing bacteria in various laboratory- and full-scale biologic processes. Researchers with a good command of primer and probe design can use qPCR as a valuable tool to study biodiversity and to develop more efficient control strategies for biologic processes.

关键词: absolute quantification     design guideline     primer     probe     real-time quantitative polymerase chain reaction (qPCR)    

基于探针图的并行型图顶点着色DNA计算模型 Article

许进, 强小利, 张凯, 张成, 杨静

《工程(英文)》 2018年 第4卷 第1期   页码 61-77 doi: 10.1016/j.eng.2018.02.011

摘要:
目前DNA 计算机研究中遇到的最大瓶颈是解空间指数爆炸问题,即随着问题规模的增大,所需要作为信息处理“数据”的DNA分子呈指数级增大。本文提出了一种新颖的图顶点着色DNA计算模型,该模型正是围绕着如何克服解空间指数爆炸问题以及如何提高运行速度而设计的。其主要贡献有:①通过如下三种方法来克服解空间指数爆炸问题:顶点颜色集的确定方法;子图分解方法以及子图中的顶点优化排序方法;②设计了一种并行型聚合酶链反应(PCR)操作技术,应用这种技术一次可以对图中多条边来删除非解,使得生物操作次数大大减少,极大地提高了运行速度。本文以一个3-着色的61 个顶点的图为例,实验表明,99% 的非可行解在构建初始解空间时就被删除,并利用DNA 自组装和并行PCR 方法,通过识别、拼接以及组装等技术得到解。由于该模型对任意61 个顶点的图是同样的操作方法,这就意味着,该模型的搜索能力可以达到O(359)。

关键词: DNA计算     图顶点着色问题     聚合酶链反应(PCR)    

A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host, Pomacea canaliculata

Fu-Rong WEI MS, Shan LV PhD, He-Xiang LIU, Ling HU, Yi ZHANG MS,

《医学前沿(英文)》 2010年 第4卷 第2期   页码 204-207 doi: 10.1007/s11684-010-0043-4

摘要: Pooling field specimens could reduce the number of assay and thus increase the efficiency in detecting and screening pathogen infections by polymerase chain reaction (PCR)-based assay. We investigated a pooling strategy in diagnosis of in Two settings of specimens were prepared, divided into portions and detected by multiplex PCR. Specimens A was 0.4490 g positive lung tissue of 28 larval nodes from four snails mixed with 1.310€g negative lung tissue from six snails and divided into 32 portions. Specimens B was 0.5448 g positive lung tissue with 26 larval nodes from two snails mixed with 1.092 g negative lung tissue from seven snails and divided into 48 portions. Repeated sampling was performed and sample size-accumulated positive rate curves were drawn. According to the sample size-accumulated positive rate curves, the appropriate sample size of the two specimens were 18 and 15, respectively, which is 0.36–0.58 to the total sample size. These test characteristics and the relevant factors to the sample size would need to be determined in much larger studies and more appropriately in field populations. The result indicates that the number of larval node is not the most important, nor the only factor to the sample size. It also implies the feasibility to detect in by pooling strategies.

关键词: Angiostrongylus cantonensis     Pomacea canaliculata     multiplex polymerase chain reaction     diagnosis     pooling    

Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

Junqin PANG, Masami MATSUDA, Masashi KURODA, Daisuke INOUE, Kazunari SEI, Kei NISHIDA, Michihiko IKE

《环境科学与工程前沿(英文)》 2016年 第10卷 第4期 doi: 10.1007/s11783-016-0846-x

摘要: Nitrogen-cycling microbial communities in municipal WWTPs were characterized. Numbers of , and genes were quantified by MPN-PCR. Diversity of whole nitrogen-cycling communities was analyzed with DNA microarray. CAS process retained diverse nitrogen cycling populations. Specific, limited populations may be dominated in nitrogen removal processes. To improve nitrogen removal performance of wastewater treatment plants (WWTPs), it is essential to understand the behavior of nitrogen cycling communities, which comprise various microorganisms. This study characterized the quantity and diversity of nitrogen cycling genes in various processes of municipal WWTPs by employing two molecular-based methods:most probable number-polymerase chain reaction (MPN-PCR) and DNA microarray. MPN-PCR analysis revealed that gene quantities were not statistically different among processes, suggesting that conventional activated sludge processes (CAS) are similar to nitrogen removal processes in their ability to retain an adequate population of nitrogen cycling microorganisms. Furthermore, most processes in the WWTPs that were researched shared a pattern:the and the bacterial genes were more abundant than the and archaeal genes, respectively. DNA microarray analysis revealed that several kinds of nitrification and denitrification genes were detected in both CAS and anaerobic-oxic processes (AO), whereas limited genes were detected in nitrogen removal processes. Results of this study suggest that CAS maintains a diverse community of nitrogen cycling microorganisms; moreover, the microbial communities in nitrogen removal processes may be specific.

关键词: DNA microarray analysis     Nitrogen cycling functional genes     Most probable number-polymerase chain reaction (MPN-PCR)     Wastewater treatment plants (WWTPs)    

标题 作者 时间 类型 操作

Effect of intestinal ischemia/reperfusion injury on leptin and orexin-A levels

LIN Ji, YAN Guangtao, HAO Xiuhua, ZHANG Kai, GAO Xiaoning, LIAO Jie

期刊论文

Abnormal expression of c-Myc in human bronchial epithelial cells malignantly transformed by anti-BPDE

FU Juan, JIANG Yiguo, CHEN Xuemin

期刊论文

Effects of 3-aminobenzamide on poly (ADP-ribose) polymerase expression, apoptosis and cell cycle progression

DU Xiang, ZHAO Hongguang, GUO Wei, GONG Shouliang, WANG Wen

期刊论文

Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

期刊论文

Advances in newly developing therapy for chronic hepatitis C virus infection

null

期刊论文

低成本PCR微反应器阵列的温度测量与控制技术

李现明,张玉林,李建田

期刊论文

Using RAPD in genotyping and transmission detection

ZHANG Tiejun, ZHOU Xiaoming, ZHANG Tao, YU Shunzhang, JIANG Qingwu, CHEN Yue

期刊论文

Expression and clinical implication of PRL-1 and PRL-3 in transitional cell carcinoma of bladder

Bin HAO, Changwei LIU, Huixiang LI

期刊论文

Seasonal microbial community shift in a saline sewage treatment plant

Qingmei YAN, Xuxiang ZHANG, Tong ZHANG, Herbert H P FANG

期刊论文

Interaction and independence on methane oxidation of landfill cover soil among three impact factors: water, oxygen and ammonium

Pinjing HE, Na YANG, Wenjuan FANG, Fan Lü, Liming SHAO

期刊论文

Cultivation of aerobic granular sludge in a conventional, continuous flow, completely mixed activated sludge system

Xi CHEN,Linjiang YUAN,Wenjuan LU,Yuyou LI,Pei LIU,Kun NIE

期刊论文

Design and use of group-specific primers and probes for real-time quantitative PCR

Juntaek LIM, Seung Gu SHIN, Seungyong LEE, Seokhwan HWANG

期刊论文

基于探针图的并行型图顶点着色DNA计算模型

许进, 强小利, 张凯, 张成, 杨静

期刊论文

A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host, Pomacea canaliculata

Fu-Rong WEI MS, Shan LV PhD, He-Xiang LIU, Ling HU, Yi ZHANG MS,

期刊论文

Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

Junqin PANG, Masami MATSUDA, Masashi KURODA, Daisuke INOUE, Kazunari SEI, Kei NISHIDA, Michihiko IKE

期刊论文