1.Establishment of a TaqMan real-time fluorescence quantitative PCR for detection of murine polyomavirus
Xueqin YIN ; Wen YUAN ; Jing WANG ; Bihong HUANG ; Dan RAO ; Miaoli WU ; Yujun ZHU ; Shengpeng FENG ; Pengju GUO ; Yu ZHANG ; Ren HUANG
Chinese Journal of Comparative Medicine 2015;(6):53-58
Objective To establish a rapid,specific and sensitive TaqMan real-time fluorescence quantitative PCR assay for detection of murine polyomavirus ( MPyV) .Methods The specific primers and TaqMan probe were designed based on genome sequence of MPyV.The primers amplified a 69 bp fragment.After optimizing the reaction system and reaction condition, the standard curve was plotted by detecting recombinant plasmid standards.The specificity, sensitivity and reproducibility of this method were evaluated.In addition, samples of lungs, spleens and feces obtained from experimentally infected mice and 86 clinical samples were used to validate the efficacy of this real-time PCR assay.Results The specificity assay showed that this assay could specifically detect MPyV and the sensitivity for MPyV was about 100 copies/well.The coefficients of variation ( CV) of both intra-assay and inter-assay were less than 1.13%.All of the samples from experimentally infected mice were positive for MPyV and 3 out of 86 clinical samples were positive by this TaqMan-PCR detection with a positive rate of 3.5%.Conclusions The real-time fluorescence quantitative TaqMan-PCR assay established in this study has high specificity, sensitivity and stability.It can be used for clinical diagnosis, routine detection and epidemiological investigation of murine polyomavirus infections.
2.Safety and efficacy between endoscopic resection and esophagectomy for T1N0 esophageal neoplasm: A systematic review and meta-analysis
Liangliang CHEN ; Jing LU ; Yujun RAO ; Yusheng SHU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(02):158-165
Objective To compare the short- and long-term efficacy of surgery and endoscopy in the treatment of early esophageal cancer by a systematic review and meta-analysis. Methods We extracted data independently from The Cochrane Library, PubMed, EMbase, Web of Science for studies comparing surgery with endoscopy from 2010 to 2020. The primary outcomes including R0 resection rate, long-term overall survival (OS), disease-specific survival (DSS), major complications, recurrence, hospital stay and cost. Meta-analysis was performed using RevMan 5.3 and Engauge Digitizer was used to extract survival curves from relevant literature, and relevant data were calculated based on statistical methods. Results A total of 17 studies involving 3 705 patients were included. It was found that patients in the surgery group had a higher R0 resection rate compared with the endoscopic group (OR=0.13, 95%CI 0.07 to 0.27, P<0.001, I2=6%). The total complications rate of resection of esophageal cancer was higher than that of the endoscopic group (OR=0.28, 95%CI 0.16 to 0.50, P<0.001, I2=68%). The length of hospitalization in the endoscopic group was obviously shorter than that in the surgery group (MD=–8.28, 95%CI –12.44 to –4.13, P<0.001, I2=96%). The distant recurrence rate (OR=0.58, 95%CI 0.24 to 1.41, P=0.230, I2=0%) and the local recurrence rate after resection (OR=1.74, 95%CI 0.66 to 4.59, P=0.260, I2=40%) in the endoscopic group was similar to those of the surgery group. There was no significant difference in 5 year-OS rate between the two groups (HR=0.86, 95%CI 0.67 to 1.11, P=0.25, I2=0%), which was subdivided into two groups: adenocarcinoma (HR=0.55, 95%CI 0.15 to 2.05, P=0.37, I2=0%) and squamous cell carcinoma (HR=0.68, 95%CI 0.46 to 1.01, P=0.06, I2=0%), showing that there was no difference between the two subgroups. There was no significant difference in the DSS rate (HR=0.72, 95%CI 0.49 to 1.05, P=0.090, I2=0%) between the two groups. The cost of the surgery group was significantly higher than that of the endoscopic group (MD=–12.97, 95%CI –18.02 to –7.92, P<0.001, I2=93%). Conclusion The evidence shows that endotherapy may be an effective treatment for early esophageal neoplasm when considering the long-term outcomes whether it is squamous or adenocarcinoma, even though it is not as effective as surgery in the short-term efficacy.