Volume 03 · Issue 01 · 2020

COVID-19目前正在中国各地爆发。政府调拨了大量的人力物力来控制这次疫情的蔓延。然而，在重要时刻，涉及使用大量消毒剂的过度预防正在远离核心爆发区的地方发生，由此引发的次生问题绝不能被忽视。

2019年底，中国湖北省武汉市出现了一种新型冠状病毒病（COVID-19），并迅速蔓延到整个中国和20多个国家。^[1,2,3,4]这种疾病是由感染严重急性呼吸综合征冠状病毒2（新型冠状病毒）引起的，冠状病毒科的成员。^[5]野生动物很可能是新型冠状病毒病毒的宿主，通过野生动物贸易传播给人类^[4,6];然而，这种新型冠状病毒的自然宿主仍然未知。需要剖析制止这一流行病的挑战，并探索进一步的战略。

A sequencing panel consisting of 50 genes was customized to reveal the potential molecular land-scapes of essential thrombocytosis, polycythemia vera, and primary myelofibrosis in Chinese patients with myeloproliferative neoplasm (MPN).

Sixty-five MPN patients (38 with essential thrombocytosis, 21 with polycythemia vera, and 6 with primary myelofibrosis), including 12 triple-negative patients, were recruited and were screened for their mutational spectrum using next-generation sequencing technology in this retrospective observational study. This study was approved by the Institutional Review Board of Changhai Hospital, Naval Military Medical University, China.

In addition to the typical driver mutations in JAK2, CALR, and MPL, pathogenic mutations in 15 other genes were frequently detected among the 65 patients with MPN. The 15 mutated genes were TET2, EZH2, ASXL1, MIR662, MLH1, MLH3, SF3B1, MSH6, BARD1, DNMT3A, KIT, MSH2, RUNX1, TP53, and NRAS in this order according to the mutational frequency detected. The average number of mutated genes was 1.2 genes per patient, while in the 12 triple-negative patients with MPN (ie, patients that lack the JAK2, CALR, or MPL mutations), at least one of the 15 pathogenic mutations was detected for each patient. Interestingly, 4 single nucleotide polymorphisms (rs4858647, rs9376092, rs58270997, rs621940) that might be correlated to individual susceptibility to myeloproliferative neoplasm were identified among the 65 patients. We also found that single nucleotide polymorphism and/or single nucleotide variation mutations occurred in multiple loci of mismatch repair-related genes, which might contribute to the development of MPN.

Our study confirms the importance of the previously known MPN relative genes and, more importantly, provides some new and potentially valuable information about mutations associated with MPNs.

Second-generation antipsychotics are widely used in mental illness, but the treatment effects and side effects are affected by single nucleotide polymorphisms (SNPs) of related genes. Quetiapine and aripiprazole are two frequently used secondgeneration antipsychotic drugs. The aim of this study was to develop two different SNP detection methods for four SNP alleles associated with the pharmacokinetics of quetiapine and aripiprazole, based on high-resolution melting (HRM) and multicolor melting curve assay (MMCA) respectively.

Whole genome DNA samples were obtained from 240 healthy people (107 females and 133 males) without genetic diseases. HRM methods were established using four kinds of specific primers and a saturated fluorescent dye. Each SNP allele with their own primers was detected in a single reaction. In the MMCA method, a multiplex polymerase chain reaction with 4 different-colored fluorescent probes was established to detect four SNP alleles in a single reaction. All experimental protocols were approved by the Ethics Committee of the Shanghai Children’s Medical Center, China (SCMC-201015) on November 22, 2010.

Two detection methods for the pharmacogenomics of quetiapine and aripiprazole, based on HRM and MMCA respectively, were established in this study. The single-target HRM method can be completed in 96 minutes, whereas the quadruplex MMCA method takes 133 minutes. It was found that the results of HRM and MMCA for the four SNP alleles had 100% coincidence with Sanger sequencing in the 240 samples.

This study developed two methods for the detection of four pharmacogenomic SNP alleles that correlated with quetiapine and aripiprazole. Both methods are rapid, cost-saving, highly accurate and potentially facilitate rational use of second-generation antipsychotics for clinical medication.

The asparagine-linked glycosylation 13 homolog (Alg13) has been identified as causative for congenital disorders of glycosylation type I with epilepsy. The aim of this study was to determine whether mice carrying a mutated version of Alg13 could be used as a model for epileptic encephalopathies or congenital disorders of glycosylation type I.

A model of epileptic encephalopathy was established in C57BL/6 mice by introducing mutations in Alg13 via the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. All surgical procedures were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University (A2016084) on October 8, 2016.

Mice with 3 different mutations, Alg13^-54nt, Alg13^-5nt and Alg13^-4nt, all of which are located in Alg13 transcript variant 1, were created. The Alg13^-5nt mice exhibited spontaneous seizures similar to patients with Alg13 mutations, suggesting that they could be used as a model for epilepsy. Western blot analysis demonstrated that Alg13^-5nt mice had lower levels of Alg13 expression than wild-type mice. Video observations showed that two of the 17 Alg13^-5nt mice had stage 5 seizures involving jumping and falling, while 12 had stage 3 seizures with head nodding.

The Alg13 mouse model provides an outstanding tool for studying epileptic encephalopathies and investigating different aspects of defects in glycosylation or other post-translational modification that cannot be assessed in patients or cell culture systems.