为建立检测F亚群禽白血病病毒(ALV-F)的SYBR Green I荧光定量PCR方法,本研究以ALV-F分离株FGD1803基因组为模板,PCR扩增其env基因,构建重组质粒pMD18-T-F作为标准品。利用该重组质粒建立了SYBR Green I荧光定量PCR方法(qPCR)的标准曲线...为建立检测F亚群禽白血病病毒(ALV-F)的SYBR Green I荧光定量PCR方法,本研究以ALV-F分离株FGD1803基因组为模板,PCR扩增其env基因,构建重组质粒pMD18-T-F作为标准品。利用该重组质粒建立了SYBR Green I荧光定量PCR方法(qPCR)的标准曲线,并经反应条件优化建立了检测ALV-F的qPCR方法。建立的标准曲线结果显示,重组质粒标准品的拷贝数与Ct值呈良好的线性关系,R^(2)为0.993,扩增效率为1.03。该方法各反应条件的优化结果为10μmol/L上下游引物各0.5μL,退火温度为55℃。特异性试验结果显示,该方法仅能特异性检测ALV-F,而对ALV-A、ALV-B、ALV-J、ALV-K、ALV-E、网状内皮增生病病毒、马立克氏病病毒、禽流感病毒、新城疫病毒、鸡支气管炎病毒等禽源病毒无交叉反应;敏感性试验结果显示,该方法对质粒标准品的最低检测限为1.16×10^(2)拷贝/μL,敏感性是常规PCR方法的100倍;以3个不同浓度重组质粒标准品为模板进行重复性试验,结果显示批内和批间重复性试验变异系数均小于5%;利用该方法与病毒培养方法分别检测30份七彩山鸡血浆样品(21份ALV阳性,9份阴性)的细胞培养物,结果显示该qPCR方法的阳性检出率为33.3%(10/30),常规PCR方法的阳性检出率为20.0%(6/30),病毒分离方法的阳性检出率为13.3%(4/30),该qPCR方法与病毒分离方法的符合率为80.0%。本研究在国内首次建立了用于ALV-F检测的qPCR方法,该方法具有良好的特异性、敏感性和重复性,可以用于临床样品的检测。为该病毒的检测提供了技术支撑。展开更多
DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the product...DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the production of hazards,limiting its practical applications.Here,we developed a DNA movable-type storage system that can utilize DNA fragments pre-produced by cell factories for data writing.In this system,these pre-generated DNA fragments,referred to herein as“DNA movable types,”are used as basic writing units in a repetitive way.The process of data writing is achieved by the rapid assembly of these DNA movable types,thereby avoiding the costly and environmentally hazardous process of de novo DNA synthesis.With this system,we successfully encoded 24 bytes of digital information in DNA and read it back accurately by means of high-throughput sequencing and decoding,thereby demonstrating the feasibility of this system.Through its repetitive usage and biological assembly of DNA movable-type fragments,this system exhibits excellent potential for writing cost reduction,opening up a novel route toward an economical and sustainable digital data-storage technology.展开更多
Background:Clinical and laboratory data were collected and analysed from patients with Ebola virus disease(EVD)in Jui Government Hospital in Freetown,Sierra Leone,where patients with EVD were received and/or treated f...Background:Clinical and laboratory data were collected and analysed from patients with Ebola virus disease(EVD)in Jui Government Hospital in Freetown,Sierra Leone,where patients with EVD were received and/or treated from October 1,2014 to March 21,2015 during the West Africa EVD outbreak.Methods:The study admitted 285 patients with confirmed EVD and followed them up till the endpoint(recovery or death).EVD was confirmed by quantitative RT-PCR assays detecting blood Ebola virus(EBOV).Results:Among the 285 lab-confirmed EVD cases in Jui Government Hospital,146 recovered and 139 died,with an overall survival rate of 51.23%.Patients under the age of 6 years had a lower survival rate(37.50%).Most non-survivors(79.86%)died within 7 days after admission and the mean hospitalization time for non-survivors was 5.56±6.11 days.More than half survivors(63.69%)turned blood EBOV negative within 3 weeks after admission and the mean hospitalization time for survivors was 20.38±7.58 days.High blood viral load(≥106 copies/ml)was found to be predictive of the non-survival outcome as indicated by the Receiver Operating Characteristic(ROC)curve analysis.The probability of patients’survival was less than 15%when blood viral load was greater than 106 copies/ml.Multivariate analyses showed that blood viral load(P=0.005),confusion(P=0.010),abdominal pain(P=0.003),conjunctivitis(P=0.035),and vomiting(P=0.004)were factors independently associated with the outcomes of EVD patients.Conclusions:Most death occurred within 1 week after admission,and patients at the age of 6 or younger had a lower survival rate.Most surviving patients turned blood EBOV negative within 1–4 weeks after admission.Factors such as high blood viral load,confusion,abdominal pain,vomiting and conjunctivitis were associated with poor prognosis for EVD patients.展开更多
BACKGROUND Gallbladder cancer is the most common malignant tumor in the biliary system,and it is characterized by high aggressiveness and an extremely poor prognosis.Current treatment for advanced gallbladder cancer r...BACKGROUND Gallbladder cancer is the most common malignant tumor in the biliary system,and it is characterized by high aggressiveness and an extremely poor prognosis.Current treatment for advanced gallbladder cancer remains unsatisfactory.Here,we report a patient with advanced gallbladder cancer who was cured by multidisciplinary treatment.CASE SUMMARY A 73-year-old male presented to our hospital with right abdominal pain for 3 d and was diagnosed with stage IVB gallbladder cancer with multiple liver metastases,peritoneum metastasis,diaphragm metastasis and lymph node metastases.The patient initially received chemotherapy,targeted therapy,125I seed implantation and immunotherapy,as there were no specific indications for radical surgery.During these palliative therapies,the level of tumor markers gradually decreased but remained higher than the normal level,lymph node metastases gradually disappeared,and liver metastasis was gradually limited to the left liver.Finally,the patient received radical surgery with left hepatectomy,radical lymphadenectomy and partial diaphragmatic resection.To date,the patient has survived for more than six years posttreatment,the levels of tumor markers are normal,and imaging examinations show no signs of tumor recurrence.CONCLUSION Currently,the prognosis of advanced gallbladder cancer remains unsatisfactory.A single treatment method is not sufficient for patients with advanced gallbladder cancer.Multidisciplinary individualized treatment is essential and should be utilized for advanced gallbladder cancer patients to further improve prognosis.展开更多
文摘为建立检测F亚群禽白血病病毒(ALV-F)的SYBR Green I荧光定量PCR方法,本研究以ALV-F分离株FGD1803基因组为模板,PCR扩增其env基因,构建重组质粒pMD18-T-F作为标准品。利用该重组质粒建立了SYBR Green I荧光定量PCR方法(qPCR)的标准曲线,并经反应条件优化建立了检测ALV-F的qPCR方法。建立的标准曲线结果显示,重组质粒标准品的拷贝数与Ct值呈良好的线性关系,R^(2)为0.993,扩增效率为1.03。该方法各反应条件的优化结果为10μmol/L上下游引物各0.5μL,退火温度为55℃。特异性试验结果显示,该方法仅能特异性检测ALV-F,而对ALV-A、ALV-B、ALV-J、ALV-K、ALV-E、网状内皮增生病病毒、马立克氏病病毒、禽流感病毒、新城疫病毒、鸡支气管炎病毒等禽源病毒无交叉反应;敏感性试验结果显示,该方法对质粒标准品的最低检测限为1.16×10^(2)拷贝/μL,敏感性是常规PCR方法的100倍;以3个不同浓度重组质粒标准品为模板进行重复性试验,结果显示批内和批间重复性试验变异系数均小于5%;利用该方法与病毒培养方法分别检测30份七彩山鸡血浆样品(21份ALV阳性,9份阴性)的细胞培养物,结果显示该qPCR方法的阳性检出率为33.3%(10/30),常规PCR方法的阳性检出率为20.0%(6/30),病毒分离方法的阳性检出率为13.3%(4/30),该qPCR方法与病毒分离方法的符合率为80.0%。本研究在国内首次建立了用于ALV-F检测的qPCR方法,该方法具有良好的特异性、敏感性和重复性,可以用于临床样品的检测。为该病毒的检测提供了技术支撑。
基金supported by the National Key Research and Development Program of China(2018YFA0900100)the Natural Science Foundation of Tianjin,China(19JCJQJC63300)Tianjin University。
文摘DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the production of hazards,limiting its practical applications.Here,we developed a DNA movable-type storage system that can utilize DNA fragments pre-produced by cell factories for data writing.In this system,these pre-generated DNA fragments,referred to herein as“DNA movable types,”are used as basic writing units in a repetitive way.The process of data writing is achieved by the rapid assembly of these DNA movable types,thereby avoiding the costly and environmentally hazardous process of de novo DNA synthesis.With this system,we successfully encoded 24 bytes of digital information in DNA and read it back accurately by means of high-throughput sequencing and decoding,thereby demonstrating the feasibility of this system.Through its repetitive usage and biological assembly of DNA movable-type fragments,this system exhibits excellent potential for writing cost reduction,opening up a novel route toward an economical and sustainable digital data-storage technology.
文摘Background:Clinical and laboratory data were collected and analysed from patients with Ebola virus disease(EVD)in Jui Government Hospital in Freetown,Sierra Leone,where patients with EVD were received and/or treated from October 1,2014 to March 21,2015 during the West Africa EVD outbreak.Methods:The study admitted 285 patients with confirmed EVD and followed them up till the endpoint(recovery or death).EVD was confirmed by quantitative RT-PCR assays detecting blood Ebola virus(EBOV).Results:Among the 285 lab-confirmed EVD cases in Jui Government Hospital,146 recovered and 139 died,with an overall survival rate of 51.23%.Patients under the age of 6 years had a lower survival rate(37.50%).Most non-survivors(79.86%)died within 7 days after admission and the mean hospitalization time for non-survivors was 5.56±6.11 days.More than half survivors(63.69%)turned blood EBOV negative within 3 weeks after admission and the mean hospitalization time for survivors was 20.38±7.58 days.High blood viral load(≥106 copies/ml)was found to be predictive of the non-survival outcome as indicated by the Receiver Operating Characteristic(ROC)curve analysis.The probability of patients’survival was less than 15%when blood viral load was greater than 106 copies/ml.Multivariate analyses showed that blood viral load(P=0.005),confusion(P=0.010),abdominal pain(P=0.003),conjunctivitis(P=0.035),and vomiting(P=0.004)were factors independently associated with the outcomes of EVD patients.Conclusions:Most death occurred within 1 week after admission,and patients at the age of 6 or younger had a lower survival rate.Most surviving patients turned blood EBOV negative within 1–4 weeks after admission.Factors such as high blood viral load,confusion,abdominal pain,vomiting and conjunctivitis were associated with poor prognosis for EVD patients.
基金Supported by National Natural Science Foundation of China,No. 81873156 and No. 82000075Liaoning Province Education Foundation,No. LZ2019051National Natural Science Foundation of Liaoning,No. 2020-BS-195
文摘BACKGROUND Gallbladder cancer is the most common malignant tumor in the biliary system,and it is characterized by high aggressiveness and an extremely poor prognosis.Current treatment for advanced gallbladder cancer remains unsatisfactory.Here,we report a patient with advanced gallbladder cancer who was cured by multidisciplinary treatment.CASE SUMMARY A 73-year-old male presented to our hospital with right abdominal pain for 3 d and was diagnosed with stage IVB gallbladder cancer with multiple liver metastases,peritoneum metastasis,diaphragm metastasis and lymph node metastases.The patient initially received chemotherapy,targeted therapy,125I seed implantation and immunotherapy,as there were no specific indications for radical surgery.During these palliative therapies,the level of tumor markers gradually decreased but remained higher than the normal level,lymph node metastases gradually disappeared,and liver metastasis was gradually limited to the left liver.Finally,the patient received radical surgery with left hepatectomy,radical lymphadenectomy and partial diaphragmatic resection.To date,the patient has survived for more than six years posttreatment,the levels of tumor markers are normal,and imaging examinations show no signs of tumor recurrence.CONCLUSION Currently,the prognosis of advanced gallbladder cancer remains unsatisfactory.A single treatment method is not sufficient for patients with advanced gallbladder cancer.Multidisciplinary individualized treatment is essential and should be utilized for advanced gallbladder cancer patients to further improve prognosis.
