Exploring the Biological and Molecular Characteristics of Resistance to Fludioxonil in Sclerotinia sclerotiorum From Soybean in China.

Sclerotinia sclerotiorum is one of the most damaging and economically important necrotrophic plant pathogens, infecting more than 400 plant species globally. Although the phenylpyrrole fungicide fludioxonil has high activity against S. sclerotiorum, reports indicate that there is also substantial potential for the development of fungicide resistance. However, the current study investigating five fludioxonil-resistant laboratory mutants found a significant fitness cost associated with fludioxonil resistance resulting in significantly (P < 0.05) reduced mycelial growth and sclerotia formation on potato dextrose agar as well as significantly (P < 0.05) lower pathogenicity on detached tomato leaves, with one mutant, LK-1R, completely losing the capacity to cause infection. In addition, all of the fludioxonil-resistant mutants had significantly (P < 0.05) increased sensitivity to osmotic stress (0.5 M of potassium chloride and 1.0 M of glucose), which is consistent with the proposed fludioxonil target sites within the high osmolarity glycerol stress response mitogen-activated protein kinase (HOG1-MAPK) signaling transduction pathway. Sequence analysis of six genes from this two-component pathway, including SsHk, SsYpd, SsSk1, SsSk2, SsPbs, and SsHog, revealed several mutations that may be associated with fludioxonil resistance. For example, six separate point mutations were found in SsHk that led to changes in the predicted amino acid sequence, including A136G, F249V, G353A, E560K, M610K, and K727R. Similarly, SsPbs had three mutations (D34G, S46L, and L337E), SsSk1 and SsYpd had two (S53G and A795V for SsSk1, and E67G and Y141H for SsYpd), and SsHog and SsSk2 had one each (V220A and S763P, respectively). To our knowledge, these constitute the first reports of amino acid changes in proteins of the HOG1-MAPK pathway being associated with fludioxonil resistance in S. sclerotiorum. This study also showed a positive cross-resistance between fludioxonil and dimethachlone and procymidone, but none with tebuconazole or carbendazim, indicating that the inclusion of tebuconazole within an integrated pest management program could reduce the risk of fludioxonil resistance developing in field populations of S. sclerotiorum and ensure the sustainable production of soybeans in China into the future.

[Four cases of acute respiratory distress syndrome patients transported with veno-venous extracorporeal membrane oxygenation].

Objective: To retrospectively review 4 cases diagnosed with pediatric acute respiratory distress syndrome (ARDS) who were transported with veno-venous (V-V) extracorporeal membrane oxygenation (ECMO) from April 2016 to March 2017. Methods: Four patients were transported to Bayi Children's Hospital Afflicted to the PLA Army General Hospital, with V-V ECMO. Their vital signs, blood-gas analysis and chest X-ray before and after transportation were compared. The length of ECMO, pediatric intensive care unit (PICU) stay and hospitalization, and the prognosis were analyzed. Results: All the four cases were transported to our hospital successfully from distances between 1 000 km to 1 210 km. The 4 cases were 4 to 6 years old with the body weight of 19 to 35 kg, of whom 3 were boys and 1 was girl. The catheters were inserted in the right jugular vein and femoral vein. The vital signs and blood-gas analysis after transportation did not change significantly compared to baseline. The length of ECMO for the four patients were 48, 754, 157 and 438 hours. They stayed in the PICU for 10, 32, 14 and 19 days, respectively. At last, 2 of them were successfully discharged from hospital without any complications; however, the other 2 died of multiple organ failure. Conclusion: Transporting ARDS patients with a satisfactory cardiac function under VV-ECMO by an experienced ECMO team is safe.

[Outcome of pediatric extracorporeal membrane oxygenation in a single center].

Objective: To investigate the application and outcome of pediatric extracorporeal membrane oxygenation (ECMO) in a single center. Methods: The clinical data of 52 pediatric patients with cardiopulmonary failure received ECMO support in Bayi Children's Hospital Affiliated to General Hospital of Beijing Military Command of PLA were collected from January 2012 to October 2016. All patients were divided into two stages by time. January 2012 to December 2014 was stage one. January 2015 to October 2016 was stage two. A retrospective analysis was done for these patients between two stages. In addition, all clinical data were compared with the data of extracorporeal life support organization (ELSO). The constituent ratio differences in different groups were tested by chi square test. Results: In 52 cases, there were 40 boys and 12 girls, aging from 1 day to 7 years, weighing from 2 to 20 kg. There were 35 cases who successfully weaned from ECMO (67%), and 25 cases were able to be discharged alive (48%). In stage one, there were 24 ECMO cases, 18 boys and 6 girls. There were 15 cases successfully weaned from ECMO (63%). Nine patients survived until discharge (38%). Complications were found in 15 cases during ECMO support (63%). In stage two, there were 28 ECMO cases, 22 were boys and 6 were girls. There were 20 cases successfully weaned from ECMO (71%). Sixteen patients survived until discharge (57%). Complications were found in 12 cases during ECMO support (43%). There was no significant difference in survival rates between two stages. However, the neonatal survival rate was higher in stage two than in stage one (71% (12/28) vs. 31% (5/24), χ(2)=5.107, P=0.038). The proportion of respiratory support was higher in stage two than in stage one (50% (14/28) vs. 21% (5/24), χ(2)=4.741, P=0.029), while the proportion of extracorporeal cardiopulmonary resuscitation (ECPR) decreased significantly (21% (6/28) vs. 67% (16/24), χ(2)=10.835, P=0.001). Application of peritoneal dialysis treatment in stage two was higher (6 vs. 0 cases, χ(2)=8.097, P=0.025). Mortality of ECMO was still higher than that of ELSO (48% (25/52) vs. 62% (34 655/55 886), χ(2)=4.281, P<0.05). The constituent ratio of different types of support varied between ECMO and ELSO patients (χ(2)=19.562, P<0.001). Conclusions: ECMO technology can provide effective support for severe cardiopulmonary failure in critically ill children. Due to the multidisciplinary nature of ECMO technology, the complexity and characteristics of pediatric patients, it takes long time to improve ECMO management and prognosis.

[Prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric acute myeloid leukemia].

Objective: To investigate the prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric patients with t (8;21) acute myeloid leukemia (AML) . Methods: The clinical features and RUNX1-RUNX1T1 transcript levels of 55 pediatric t (8;21) AML patients, newly diagnosed from Jan. 2010 to Apr. 2016, were analyzed retrospectively. The relationship between the minimal residual disease (MRD) and prognosis was analysed by dynamic monitoring of RUNX1-RUNX1T1 transcript levels using real-time quantitative PCR (RQ-PCR) technology. Results: The RUNX1-RUNX1T1 transcript levels in bone marrow cells at diagnosis was not related to relapse. After one course of induction therapy, patients with a more than 2 Log reduction of RUNX1-RUNX1T1 transcript levels (>2 Log) had lower 5 years cumulative incidence of relapse (CIR) [ (24.3±8.4) % vs (52.6±9.7) %, χ(2)=9.046, P=0.003], relapse-free survival (RFS) [ (71.6±12.7) % vs (48.1±13.2) %, χ(2)=5.814, P=0.016], and better overall survival (OS) [ (76.9±12.5) % vs (48.9±14.7) %, χ(2)=6.346, P=0.012], compared to patients with a less than 2 Log reduction (a<2 Log) . Multivariate Cox survival analysis suggested that a>2 Log reduction in RUNX1-RUNX1T1 transcript levels after a course of induction therapy was an independent prognostic factor for RFS (HR=0.263, 95%CI 0.081-0.851, P=0.026) and OS (HR=0.214, 95% CI 0.057-0.808, P=0.023) . During consolidation therapy and follow-up period, molecular relapse of 16 cases and hematologic relapse of 13 cases were identified by continuous dynamic monitoring of RUNX1-RUNX1T1 transcript levels, with a median interval of 4.0 (1.5-5.8) months from the molecular relapse to hematologic relapse. 2 cases of molecular relapse who received timely allogeneic hematopoietic stem cell transplantation did not experience hematologic relapse. Conclusion: Dynamic monitoring RUNX1-RUNX1T1 transcript levels by RQ-PCR technique can subdivide patients into relatively low and high risk group, early screen patients at high risk of relapse and provide a scientific basis for precision stratification and risk-adapted therapy for pediatric t (8;21) AML children.

SPE-UPLC-MS/MS for the determination of phthalate monoesters in rats urine and its application to study the effects of food emulsifier on the bioavailability of priority controlling PAEs.

This research was mainly focused on the effects of food emulsifier on the bioavailability of six priority controlling phthalate acid esters (PAEs) for the further accurate assessment of their toxic effects, using the corresponding phthalate acid monoesters (PAMEs) in rats urine as biomarkers. Glycerin monostearate was chosen as typical food emulsifier. A method was established to determine PAMEs in urine from rats either in experimental group (integrated gavaged with glycerin monostearate and PAEs) or in control group (gavaged with PAEs only), by using solid-phase extraction (SPE) coupled with ultra performance liquid chromatography tandem mass spectrometry (SPE-UPLC-MS/MS). Extraction recoveries were more than 75% for all the PAMEs; the calibration curve was linear in the range of 1.0-1000.0ng/mL with R(2)>0.995; the limits of detection (LOD) were 0.30ng/mL-0.50ng/mL. In addition, by analysing quality control (QC) urine samples in 3 days, it showed that the method was precise and accurate, for the intra-day and inter-day RSD within 16%, and the accuracy more than 82%. Internal exposure amount of all PAEs in experimental group was significantly higher than that in control group with p values of less than 0.05 except for butyl benzyl phthalates (BBP) (P=0.07). The bioavailability of all PAEs ranged from 5.03% to 109.35% with the presence of food emulsifiers glycerin monostearate, observably higher than that without glycerin monostearate (1.12% to 54.39%). It indicated that food emulsifiers increased the bioavailability of PAEs and may lead to potential food safety risk, which should bring awareness and be further studied.