OBJECTIVEDevelopment and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.

METHODSThe VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.

RESULTSIn this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.

CONCLUSIONThe results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Polyomavirus ; isolation & purification ; Real-Time Polymerase Chain Reaction ; methods ; Respiratory Tract Infections ; virology ; Sputum ; virology

Objective:To understand the radioactivity levels of water source in five key cities of Chengdu, Yibin, Mianyang, Guangyuan, Leshan in Sichuan province, and evaluate the radiation safety of these water sources.Methods:From 2016 to 2020, the measuring site selection, sampling and analysis were made for measuring the radioactivity levels of total alpha, total beta, 90Sr, 137Cs, 226Ra, U, Th in drinking water sources in these five cities. Reference was made, respectively, to the national standards Determination of total α radioactivity concentration in water thick source method (EJ/T 1075-1998), Determination of total β radioactivity in water evaporation method (EJ/T 900-1994), Radiochemical analysis method of strontium-90 in water and biological samples ash (HJ 815-2016), radiochemical analysis of cesium-137 in the ash of water and biological samples (HJ 816-2016), Analysis and determination of radium-226 in water (GB 11214-1989), Analysis of trace uranium in environmental samples -3 laser fluorescence law (HJ 840-2017), and Food safety in the test of radioactive substances in food determination of natural thorium and uranium (GB 14883.7-2016). Results:The activity concentration ranges from 0.011 to 0.076 Bq/L for total alpha, 0.027 to 0.098 Bq/L for total beta, 0.21 to 0.45 mBq/L for 137Cs, 1.0 to 2.4 mBq/L for 90Sr, 6.1 to 16.0 mBq/L for 226Ra, 0.06 to 0.21 μg/L for Th, and 0.73 to 3.30 μg/L for U. There were no significant differences in the concentrations of total alpha, total beta, 90Sr, 137Cs, 226Ra, U and Th in different water sources and different years ( P>0.05). There were no significant differences in the concentrations of total alpha, total beta, 90Sr, 137Cs and 226Ra during different water periods ( P>0.05). There were statistical differences in U activity concentration in different water periods ( H=16.53, P<0.05), and statistical differences in Th activity concentration in different water periods ( H=15.61, P<0.05). Conclusions:The survey showed that the total alpha, total beta, 90Sr, 137Cs, 226Ra, U and Th radioactivity levels in drinking water sources in Chengdu, Yibin, Mianyang, Guangyuan and Leshan of Sichuan province were at the same level as the relevant data in the Natural environmental radioactivity level in China published by the State Environmental Protection Bureau in 1995. All of there values were within the data range released by the 2013-2021 National Radiation Environment Monitoring Quality Report issued by the Radiation Environment Monitoring Technology Center of the Ministry of Ecology and Environment. The radioactivity level in water sources in the five key cities was at a safe level.

Objective To introduce the technique of interictal 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) examination and explore the value of 18F-FDG-PET in the localization of epileptogenic focus of temporal lobe epilepsy (TLE) confirmed by surgical result.Methods Clinical data were retrospectively analyzed in 82 TLE patients having received interictal 18F-FDG-PET preoperative evaluation and got EngleⅠ grade epileptic surgical outcome, and the sensitivity and specialty of interictal 18F-FDG-PET were compared with those of MRI and scalp video-EEG. Results Epileptogenic foci showed hypometabolism on 18F-FDG-PET, and the hypometabolism zones were localized in ipsilateral temporal lobe in 68 cases,beyond ipsilateral temporal lobe in 9 cases; the other 5 had no hypometabolism zone. Accuracy rate of localization of epileptogenic foci by interictal 18F-FDG-PET was 82.9% (68/82), significantly higher than that by MRI or EEG(P＜0.05).77.4%(41/53)epileptogenic foci where MRI showed negative and 75%(15/20)where EEG with imbedded electrode was applied were precisely localized by 18F-FDG-PET. The accuracy was higher in the cases with positive pathological result than in the ones with negative result. Conclusions Interictal 18F-FDG-PET possesses excellent sensitivity and specialty in preopemtive assessment for TLE, and is of good value in the localization of epileptogenic focus where MRI shows negative or invasive electrophysiologic monitoring is needed.Rational application may raise the accuracy rate.

This study was aimed to enhance clinical understanding the effect of nilotinib on CML patients with V299L mutation who were resistant to imatinib. Bone marrow specimens from 2 cases of CML with V299L mutation were collected before and after the treatment with nilotinib. ABL mutation was detected by nested reverse transcription polymerase chain reaction (PCR) followed by direct sequencing. The clinical characteristics of the two cases were analyzed. The results showed that both cases were resistant to imatinib presented V299L mutation. Out of them 1 case achieved complete haematological response (CHR) after treatment with nilotinib for 6 months and another case abstained obvious molecular response after using nilotinib for 7 month. V299L mutation of both cases was turned into negative after the treatment with nilotinib. It is concluded that the nilotinib can safely and effectively override tyrosine kinase inhibitor (TKI) resistance mediated by the V299L mutation. The safety and efficacy of nilotinib for treatment of CML patients with TKI resistance and V299L mutation are satisfactory.
Adult ; Aged ; Benzamides ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; Male ; Mutation ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; therapeutic use