Objective: This study aims to establish a method for highly parallel multiplexed detection of genetic mutations in Chinese lung cancer samples through Agena i PLEX chemistry and matrix-assisted laser desorption ioniza...Objective: This study aims to establish a method for highly parallel multiplexed detection of genetic mutations in Chinese lung cancer samples through Agena i PLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis on Mass ARRAY mass spectrometry platform.Methods: We reviewed the related literature and data on lung cancer treatments. We also identified 99 mutation hot spots in 13 target genes closely related to the pathogenesis, drug resistance, and metastasis of lung cancer. A total of 297 primers, composed of99 paired forward and reverse amplification primers and 99 matched extension primers, were designed using Assay Design software. The detection method was established by analyzing eight cell lines and six lung cancer specimens. The proposed method was then validated through comparisons by using a Lung Carta^(TM) kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases.Results: The proposed method was able to detect multiplex genetic mutations in lung cancer cell lines. This finding was consistent with the observations on previously reported mutations. The proposed method can also detect such mutations in clinical lung cancer specimens. This result was consistent with the observations with Lung Carta^(TM) kit. However, an FGFR2 mutation was detected only through the proposed method. The measured sensitivity and specificity were 100% and 96.3%, respectively.Conclusions: The proposed Mass ARRAY technology-based multiplex method can detect genetic mutations in Chinese lung cancer patients. Therefore, the proposed method can be applied to detect mutations in other cancer tissues.展开更多
BACKGROUND Data that assess maternal and infant outcomes in hepatitis C virus(HCV)-infected mothers are limited.AIM To investigate the frequency of complications and the associated risk factors.METHODS We performed a ...BACKGROUND Data that assess maternal and infant outcomes in hepatitis C virus(HCV)-infected mothers are limited.AIM To investigate the frequency of complications and the associated risk factors.METHODS We performed a cohort study to compare pregnancy and fetal outcomes of HCVviremic mothers with those of healthy mothers.Risk factors were analyzed with logistic regression.RESULTS Among 112 consecutive HCV antibody-positive mothers screened,we enrolled 79 viremic mothers.We randomly selected 115 healthy mothers from the birth registry as the control.Compared to healthy mothers,HCV mothers had a significantly higher frequency of anemia[2.6%(3/115)vs 19.0%(15/79),P<0.001]during pregnancy,medical conditions that required caesarian section[27.8%(32/115)vs 48.1%(38/79),P=0.004],and nuchal cords[9.6%(11/115)vs 34.2%(27/79),P<0.001].In addition,the mean neonatal weight in the HCV group was significantly lower(3278.3±462.0 vs 3105.1±459.4 gms;P=0.006),and the mean head circumference was smaller(33.3±0.6 vs 33.1±0.7 cm;P=0.03).In a multivariate model,HCV-infected mothers were more likely to suffer anemia[adjusted odds ratio(OR):18.1,95%confidence interval(CI):4.3-76.6],require caesarian sections(adjusted OR:2.6,95%CI:1.4-4.9),and have nuchal cords(adjusted OR:5.6,95%CI:2.4-13.0).Their neonates were also more likely to have smaller head circumferences(adjusted OR:2.1,95%CI:1.1-4.3)and lower birth weights than the average(≤3250 gms)with an adjusted OR of 2.2(95%CI:1.2-4.0).The vertical transmission rate was 1%in HCV-infected mothers.CONCLUSION Maternal HCV infections may associate with pregnancy and obstetric complications.We demonstrated a previously unreported association between maternal HCV viremia and a smaller neonatal head circumference,suggesting fetal growth restriction.展开更多
Mitochondria play a kay role in various cell processes including ATP production, Ca^2＋ homeostasis, reactive oxygen species （ROS） generation, and apoptosis. The selective removal of impaired mitochondria by autopha...Mitochondria play a kay role in various cell processes including ATP production, Ca^2＋ homeostasis, reactive oxygen species （ROS） generation, and apoptosis. The selective removal of impaired mitochondria by autophagosome is known as mitophagy. Cerebral ischemia is a common form of stroke caused by insuf- ficient blood supply to the brain. Emerging evidence suggests that mitophagy plays important roles in the pathophysiological process of cerebral ischemia. This review focuses on the relationship between ischemic brain injury and mitophagy. Based on the latest research, it describes how the signaling pathways of mitophagy appear to be involved in cerebral ischemia.展开更多
Background： Drug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （TKIs） are continuously being disco...Background： Drug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （TKIs） are continuously being discovered. We aimed to establish a novel method for highly parallel multiplexed detection of genetic mutations related to EGFR TKl-resistant lung cancer using Agena iPLEX chemistry and niatrix-assisted laser desorption ionization time-of-flight analysis on the MassARRAY mass spectrometry platform. Methods： A review of the literature revealed 60 mutation hotspots in seven target genes （EGFR, KRAS, PIK3CA, BRAF, ERBB2, NRAS, and BIM） that are closely related to EGFR TKI resistance to lung cancer. A total of 183 primers comprised 61 paired forward and reverse amplification primers, and 61 matched extension primers were designed using Assay Design Software. The detection method was established by analyzing nine cell lines, and by comparison with LungCartaTM kit in ten lung cancer specimens. EGFR, KRAS. and BIM genes in all cell lines and clinical samples were subjected to Sanger sequencing for confirming reproducibility. Results： Our data showed that designed panel was a high-throughput and robust tool, allowing genotyping for sixty hotspots in the same run. Moreover, it made efficient use of patient diagnostic samples for a more accurate EGFR TKIs resistance analysis. The proposed method could accurately detect mutations in lung cancer cell lines and clinical specimens, consistent with those obtained by the LungCartaTM kit and Sanger sequencing. We also established a method for detection of large-fragment deletions based on single-base extension technology of MassARRAY platform. Conclusions： We established an effective method for high-throughput detection of genetic mutations related to EGFR TKI resistance based on the MassARRAY platform, which could provide more accurate information for overcoming cancers with de novo or acquired resistance to EGFR-targeted therapies.展开更多
基金supported by the Special Fund for Research in the Public Interest from the National Health and Family Planning Commission of PRC (Grant No. 201402031)the Key Lab System Project of the Guangdong Science and Technology Department (Grant No. 2012A061400006)the Special Fund for Research in the Public Interest and Capacity Building from the Guangdong Science and Technology Department (Grant No. 2014A020212225)
文摘Objective: This study aims to establish a method for highly parallel multiplexed detection of genetic mutations in Chinese lung cancer samples through Agena i PLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis on Mass ARRAY mass spectrometry platform.Methods: We reviewed the related literature and data on lung cancer treatments. We also identified 99 mutation hot spots in 13 target genes closely related to the pathogenesis, drug resistance, and metastasis of lung cancer. A total of 297 primers, composed of99 paired forward and reverse amplification primers and 99 matched extension primers, were designed using Assay Design software. The detection method was established by analyzing eight cell lines and six lung cancer specimens. The proposed method was then validated through comparisons by using a Lung Carta^(TM) kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases.Results: The proposed method was able to detect multiplex genetic mutations in lung cancer cell lines. This finding was consistent with the observations on previously reported mutations. The proposed method can also detect such mutations in clinical lung cancer specimens. This result was consistent with the observations with Lung Carta^(TM) kit. However, an FGFR2 mutation was detected only through the proposed method. The measured sensitivity and specificity were 100% and 96.3%, respectively.Conclusions: The proposed Mass ARRAY technology-based multiplex method can detect genetic mutations in Chinese lung cancer patients. Therefore, the proposed method can be applied to detect mutations in other cancer tissues.
基金Supported by The Ministry of Science and Technology of China for the National Five-Year Key Projects in Infectious Diseases,No. 2015ZX10004801
文摘BACKGROUND Data that assess maternal and infant outcomes in hepatitis C virus(HCV)-infected mothers are limited.AIM To investigate the frequency of complications and the associated risk factors.METHODS We performed a cohort study to compare pregnancy and fetal outcomes of HCVviremic mothers with those of healthy mothers.Risk factors were analyzed with logistic regression.RESULTS Among 112 consecutive HCV antibody-positive mothers screened,we enrolled 79 viremic mothers.We randomly selected 115 healthy mothers from the birth registry as the control.Compared to healthy mothers,HCV mothers had a significantly higher frequency of anemia[2.6%(3/115)vs 19.0%(15/79),P<0.001]during pregnancy,medical conditions that required caesarian section[27.8%(32/115)vs 48.1%(38/79),P=0.004],and nuchal cords[9.6%(11/115)vs 34.2%(27/79),P<0.001].In addition,the mean neonatal weight in the HCV group was significantly lower(3278.3±462.0 vs 3105.1±459.4 gms;P=0.006),and the mean head circumference was smaller(33.3±0.6 vs 33.1±0.7 cm;P=0.03).In a multivariate model,HCV-infected mothers were more likely to suffer anemia[adjusted odds ratio(OR):18.1,95%confidence interval(CI):4.3-76.6],require caesarian sections(adjusted OR:2.6,95%CI:1.4-4.9),and have nuchal cords(adjusted OR:5.6,95%CI:2.4-13.0).Their neonates were also more likely to have smaller head circumferences(adjusted OR:2.1,95%CI:1.1-4.3)and lower birth weights than the average(≤3250 gms)with an adjusted OR of 2.2(95%CI:1.2-4.0).The vertical transmission rate was 1%in HCV-infected mothers.CONCLUSION Maternal HCV infections may associate with pregnancy and obstetric complications.We demonstrated a previously unreported association between maternal HCV viremia and a smaller neonatal head circumference,suggesting fetal growth restriction.
基金This work was funded by the Faculty Research Grant of Hong Kong Baptist University （FRG2/15-16/022） and the Guandong Natural Science Foundation （2014A030313766 and 2016A030313008）.
文摘Mitochondria play a kay role in various cell processes including ATP production, Ca^2＋ homeostasis, reactive oxygen species （ROS） generation, and apoptosis. The selective removal of impaired mitochondria by autophagosome is known as mitophagy. Cerebral ischemia is a common form of stroke caused by insuf- ficient blood supply to the brain. Emerging evidence suggests that mitophagy plays important roles in the pathophysiological process of cerebral ischemia. This review focuses on the relationship between ischemic brain injury and mitophagy. Based on the latest research, it describes how the signaling pathways of mitophagy appear to be involved in cerebral ischemia.
文摘Background： Drug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （TKIs） are continuously being discovered. We aimed to establish a novel method for highly parallel multiplexed detection of genetic mutations related to EGFR TKl-resistant lung cancer using Agena iPLEX chemistry and niatrix-assisted laser desorption ionization time-of-flight analysis on the MassARRAY mass spectrometry platform. Methods： A review of the literature revealed 60 mutation hotspots in seven target genes （EGFR, KRAS, PIK3CA, BRAF, ERBB2, NRAS, and BIM） that are closely related to EGFR TKI resistance to lung cancer. A total of 183 primers comprised 61 paired forward and reverse amplification primers, and 61 matched extension primers were designed using Assay Design Software. The detection method was established by analyzing nine cell lines, and by comparison with LungCartaTM kit in ten lung cancer specimens. EGFR, KRAS. and BIM genes in all cell lines and clinical samples were subjected to Sanger sequencing for confirming reproducibility. Results： Our data showed that designed panel was a high-throughput and robust tool, allowing genotyping for sixty hotspots in the same run. Moreover, it made efficient use of patient diagnostic samples for a more accurate EGFR TKIs resistance analysis. The proposed method could accurately detect mutations in lung cancer cell lines and clinical specimens, consistent with those obtained by the LungCartaTM kit and Sanger sequencing. We also established a method for detection of large-fragment deletions based on single-base extension technology of MassARRAY platform. Conclusions： We established an effective method for high-throughput detection of genetic mutations related to EGFR TKI resistance based on the MassARRAY platform, which could provide more accurate information for overcoming cancers with de novo or acquired resistance to EGFR-targeted therapies.
