The web-based multiplex PCR primer design software Ultiplex and the associated experimental workflow: up to 100- plex multiplicity

Background A large number of variants have been employed in various medical applications, such as providing medication instructions, disease susceptibility testing, paternity testing, and tumour diagnosis. A high multiplicity PCR will outperform other technologies because of its lower cost, reaction time and sample consumption. To conduct a multiplex PCR with higher than 100 plex multiplicity, primers need to be carefully designed to avoid the formation of secondary structures and nonspecific amplification between primers, templates and products. Thus, a user-friendly, highly automated and highly user-defined web-based multiplex PCR primer design software is needed to minimize the work of primer design and experimental verification. Results Ultiplex was developed as a free online multiplex primer design tool with a user-friendly web-based interface (http://ultiplex.igenebook.cn). To evaluate the performance of Ultiplex, 294 out of 295 (99.7%) target primers were successfully designed. A total of 275 targets produced qualified primers after primer filtration, and 271 of those targets were successfully clustered into one compatible PCR group and could be covered by 108 primers. The designed primer group stably detected the rs28934573(C > T) mutation at lower than a 0.25% mutation rate in a series of samples with different ratios of HCT-15 and HaCaT cell line DNA. Conclusion Ultiplex is a web-based multiplex PCR primer tool that has several functions, including batch design and compatibility checking for the exclusion of mutual secondary structures and mutual false alignments across the whole genome. It offers flexible arguments for users to define their own references, primer Tm values, product lengths, plex numbers and tag oligos. With its user-friendly reports and web-based interface, Ultiplex will provide assistance for biological applications and research involving genomic variants.

Primary and Metastatic Brain Tumours Assessed with the Brain and Torso [18F]FDG PET/CT Study Protocol—10 Years of Single-Institutional Experiences

According to the international societies’ recommendations, the 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography/computed tomography ([18F]FDG PET/CT) technique should not be used as the method of choice in brain tumour diagnosis. Therefore, the brain region can be omitted during standard [18F]FDG PET/CT scanning. We performed comprehensive literature research and analysed results from 14,222 brain and torso [18F]FDG PET/CT studies collected in 2010–2020. We found 131 clinically silent primary and metastatic brain tumours and 24 benign lesions. We concluded that the brain and torso [18F]FDG PET/CT study provides valuable data that may support therapeutic management by detecting clinically silent primary and metastatic brain tumours.

‘Hybrid’ oncocytoma: collecting duct (Bellini) carcinoma—the peril from this extremely rare intratumoural coexistence

We presented an extremely rare entity of ‘hybrid’ oncocytoma and collecting duct (Bellini) carcinoma. The intratumoural coexistence of benign and malignant cells may lead to false diagnosis and suboptimal treatment of an aggressive tumour. Diagnosis may be challenging if only based on imaging modalities. Even the established value of targeted renal biopsy may be questioned in such scarce cases. Consequently, active surveillance for small renal tumours shall not considered a widely safe management.

Testicular Sertoli cell tumour and potentially testicular Leydig cell tumour are features of DICER1 syndrome

DICER1 syndrome is a rare paediatric autosomal dominant inherited disorder predisposing to various benign and malignant tumours. It is caused by a germline pathogenic variant in DICER1, and the second hit for tumour development is usually a missense hotspot pathogenic variant in the DICER1 ribonuclease IIIb domain. While DICER1 predisposing variants account for about 60% of ovarian Sertoli-Leydig cell tumours, no DICER1-related testicular stromal tumours have been described. Here we report the first two cases of testicular stromal tumours in children carrying a DICER1 germline pathogenic variant: a case of Sertoli cell tumour and a case of Leydig cell tumour diagnosed at 2 and 12 years of age, respectively. A somatic DICER1 hotspot pathogenic variant was detected in the Sertoli cell tumour. This report extends the spectrum of DICER1-related tumours to include testicular Sertoli cell tumour and potentially testicular Leydig cell tumour. Diagnosis of a testicular Sertoli cell tumour should prompt DICER1 genetic testing so that patients with a DICER1 germline pathogenic variant can benefit from established surveillance guidelines. DICER1 genetic evaluation may be considered for testicular Leydig cell tumour. Our findings suggest that miRNA dysregulation underlies the aetiology of some testicular stromal tumours.

An Active and Low-Cost Microwave Imaging System design for Detection of Breast Cancer Using Back Scattered Signal

: A defected ground antenna with dielectric reflector is designed and investigated for breast tumour diagnosis. Ultra-wide band resonance (3.1 to 10.6 GHz) is achieved by etching two slots and adding a narrow vertical strip in a patch antenna. A high dielectric constant substrate is added below the antenna, which shows remarkable effect on performance. Antenna performance is verified experimentally on an artificially fabricated breast tissue and tumour. Malignant tissue has different dielectric properties than the normal tissue, that causes deviation in the scattered antenna power. Average value of backscattered signal variation and ground penetrating radar (GPR) algorithm is used to localize the tumour of radius 4mm in breast tissue.