Objective To study the FMS-like tyrosine kinase-3 (FLT3) gene, NPM1 gene and c-kit gene mutations in acute myeloid leukemia (AML) by extracting DNA from the storage of bone marrow slides, and to investigate the relationship between the three gene mutations and clinical features in AML. Methods The bone marrow slides of 55 patients diagnosed with AML were enrolled in this study. The PCR, DNA sequencing and molecular cloning were used to detect and analyse the FLT3-ITD, NPM1 and c-kit gene mutations. Patients' remission, progression and survival time were also recorded. Results The DNA was successfully extracted from the bone marrow slides with -20 ℃ frozen storage without Wright stained, chemically fixed, and room temperature storage Wright stained discoloured by phenol ∶ chloroform ∶ isoamyl alcohol method, which can be used in PCR, direct sequencing and molecular cloning sequencing analysis. 10 of the 55 cases (18.2 %) were FLT3-ITD positive, including 9 cases with heterozygous mutations and 1 case with homozygous mutation. FLT3-ITD positive group had lower complete remission (CR) rate, shorter event-free survival (EFS) time and overall survival (OS) time than the negative group (P< 0.05). 9 of the 55 cases (16.4 %) had NPM1 heterozygous gene mutations, all belonging to type A. The EFS rate of the patients with NPM1 mutation was higher in 10 months and the OS rate was higher in 19 months (P< 0.05). 3 of 9 NPM1 mutations patients were FLT3-ITD positive. The CR rates of the four groups after initial remission induction therapy in order were NPM1+FLT3-ITD-, NPM1-FLT3-ITD-, NPM1-FLT3-ITD+, NPM1+FLT3-ITD+(P<0.05). Besides, NPM1-FLT3-ITD+was a risk factor affecting the OS (RR=1.250, P=0.005). 2 of the 55 cases (3.6 %) had c-kit gene mutations, namely mutant D816H and mutant D816V. The c-kit gene mutations were not found in patients with FLT3-ITD and NPM1 mutations. Conclusions The FLT3-ITD mutation is a poor prognosis molecular marker in AML, and NPM1 mutation is a good factor for the prognosis. NPM1-FLT3-ITD+is a risk factor affecting OS. The rate of c-kit gene mutation is low in AML, without the overlap of FLT3 and NPM1 mutations.

Objective:To disclose the molecular genetic differences of Culex flavivirus among mosquitoes in Gansu province in 2011 and 2019.Methods:Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to obtain the nucleotide sequences of Culex flavivirus genomes from mosquitoes in Gansu province in 2011 and 2019, and viral molecular biology and bioinformatics method were used to analyze the genetic differences of the viruses.Results:Nucleotide sequences of 10 strains of Culex flavivirus were obtained, including 8 strains (all from Culex pipiens pallens) obtained in 2011 and 2 strains ( from Culex tritaeniorhynchus and Anopheles sinensis) in 2019. Homology analysis of nucleotide and amino acid sequences of virus E gene showed that the nucleotide sequence similarity and amino acid similarity of viruses isolated from Gansu in 2019 and 2011 ranged from 98.3%-100% and 95.4%-97.3%, respectively. Phylogenetic analysis of Culex flavivirus E gene sequence showed that two strains of Culex flavivirus isolated in Gansu province in 2019 (GS1975 and GS1976) and eight strains of Culex flavivirus isolated in 2011 all belonged to group B of genotype 1 of Culex flavivirus. Further analysis found that GS1975 virus isolated in 2019 was in a common evolutionary cluster with viruses isolated from Liaoning (2010 and 2011) and Inner Mongolia (2018), while GS1976 virus isolated in 2019 formed a coevolutionary cluster with viruses isolated from Inner Mongolia (2018) and Gansu (2011). Conclusions:Although both Culex flaviviruses isolated in Gansu province in 2011 and 2019 are genotype 1 virus, the two viruses isolated in 2019 distributed in two different evolutionary clusters, suggesting that the local mosquito virus genome changes over time, therefore, long-term monitoring of molecular differences is needed to carry out.

Objective:To perform virological and molecular biological identification of the virus (SX1943) isolated from specimens of Culex tritaeniorhynchus in Shaanxi province in 2019. Methods:Mosquito ground fluid was inoculated into tissue culture cells, and isolates were made virological and molecular genetic analysis.Results:A virus isolate (SX1943) was obtained from a specimen of Culex tritaeniorhynchus collected in Shaanxi province in 2019, which developed significant cytopathic effect (CPE) on day 3 after inoculation into C6/36 cells, manifested as cell pyknosis, aggregation and shedding, and the isolate could be stably passaged. However, no significant CPE was observed after three consecutive passages in BHK-21 cells. Electron microscopy (negative staining) of C6/36 cells inoculated with SX1943 virus showed a large number of round virus particles, about 25 nm in diameter. The result of amplification and sequencing of the SX1943 viral genome showed that the total length of the viral genome sequence was 3 594 nt, encoding a total of three proteins, which were non-structural proteins (NS1 and NS2) and capsid proteins (VP), and the gene lengths of the three proteins were 2 376 nt, 1 098 nt and 1 136 nt, respectively. Phylogenetic analysis of the virus revealed that the SX1943 virus were in the same evolutionary clade as Culex pipiens pallens Densovirus (CppDNV) in the genus Brevidensovirus in the subfamily Densovirus, and the above result suggested that SX1943 virus was CppDNV. Conclusions:CppDNV was isolated from Culex tritaeniorhynchus in Shaanxi province.

BACKGROUND: Intracerebral hemorrhage (ICH) is one of the most severe complications during veno-venous extracorporeal membrane oxygenation (VV-ECMO). This study aimed to determine the risk factors for ICH and mortality in such patients. METHODS: We analyzed the clinical data of 77 patients who received VV-ECMO due to severe respiratory failure from July 2013 to May 2019 at China-Japan Friendship Hospital. Demographical data, laboratory indices, imaging characteristics, and other clinical information were collected. Multivariable logistic regression analyses were performed to identify risk factors for ICH and mortality. RESULTS: Of 77 patients, 11 (14.3%) suffered from ICH, and 36 (46.8%) survived. The survival rate was significantly lower (18.2% [2/11] vs. 51.5% [34/66], P = 0.040) in patients with ICH than in those without ICH. Multivariable analysis revealed that factors independently associated with ICH were diabetes mellitus (adjusted odds ratio [aOR]: 12.848, 95% confidence interval [CI]: 1.129-146.188, P = 0.040) and minimum fibrinogen during ECMO (aOR: 2.557, 95% CI: 1.244-5.252, P = 0.011). Multivariable analysis showed that factors independently associated with mortality were acute hepatic failure during ECMO (aOR: 9.205, 95% CI: 1.375-61.604, P = 0.022), CO2 retention before ECMO (aOR: 7.602, 95% CI: 1.514-38.188, P = 0.014), and minimum platelet concentration during ECMO (aOR: 0.130, 95% CI: 0.029-0.577, P = 0.007). CONCLUSIONS Diabetes mellitus and minimum fibrinogen concentration during ECMO are risk factors for ICH in patients with severe respiratory failure managed using VV-ECMO. This indicated that anticoagulants use and nervous system monitoring should be performed more carefully in patients with diabetes when treated with VV-ECMO due to severe respiratory failure.
Adult ; Anticoagulants ; Extracorporeal Membrane Oxygenation ; Humans ; Intracranial Hemorrhages ; Respiratory Insufficiency/therapy* ; Retrospective Studies ; Risk Factors