Genetic Variation of Magnaporthe oryzae Population in Hunan Province.

Studies on the population structure and variation of Magnaporthe oryzae in fields are of great significance for the control of rice blast disease. In this study, a total of 462 isolates isolated from different areas of Hunan Province in 2016 and 2018 were analyzed for their population structure and variation tendency. The results showed that from 2016 to 2018, the concentration of fungal races of M. oryzae increased and the diversity decreased; furthermore, 218 isolates in 2016 belonged to ZA, ZB, ZC, ZE, ZF and ZG, with a total of 6 groups and 29 races, in which the dominant-population ZB group accounted for 66.2%; meanwhile, in 2018, 244 isolates were classified into 4 groups and 21 races, including ZA, ZB, ZC and ZG, in which the dominant-population ZB group accounted for 72.54%. In 2018, isolates of ZD, ZE and ZF populations were absent, and the number of total races and isolates of the ZA and ZC groups decreased. Fungal pathogenicity was identified, with 24 monogenic lines (MLs) carrying 24 major R genes. The resistance frequency of R genes to fungal isolates in 2018 decreased significantly, in which except Pikm was 64.5%, the other monogenic lines were less than 50%. Rep-PCR analysis for isolates of Guidong in Hunan also showed that fungal diversity decreased gradually. The influence of R genes on fungal variation was analyzed. The pathogenicity of isolates purified from Xiangwanxian 11 planted with monogenic lines was significantly more enhanced than those without monogenic lines. All the results indicated that in recent years, the fungal abundance in Hunan has decreased while fungal pathogenicity has increased significantly. This study will greatly benefit rice-resistance breeding and the control of rice blast disease in Hunan Province.

Alternaria alternata F3, a Novel Taxol-Producing Endophytic Fungus Isolated from the Fruits of Taxus cuspidata: Isolation, Characterization, Taxol Yield Improvement, and Antitumor Activity.

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4 µg L-1 by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1 g L-1 sodium acetate, 0.25 g L-1 salicylic acid, and 0.00125 g L-1 silver nitrate and inoculum size 2%, and incubated at 28 °C and 150 rpm for 8 days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC50 values of 3.98 µg mL-1 and 0.35 µg mL-1. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

FERONIA-like receptor 1-mediated calcium ion homeostasis is involved in the immune response.

Calcium (Ca2+) is the most abundant divalent cation in plants, and cellular levels of Ca2+, which functions as a nutrient and secondary messenger, play a critical role in plant immunity. In the present study, we found that FERONIA-like receptor 1 (FLR1) positively regulates Magnaporthe oryzae resistance and that expression of FLR1 is strongly induced in response to Ca2+ deficiency. In addition, the Ca content in the shoots of flr1 was lower than that in wild-type, and the M. oryzae-sensitive phenotype of the flr1 mutant was not rescued by exogenous application of Ca2+. Moreover, RNA sequencing revealed 2,697 differentially expressed genes (DEGs) in the flr1 mutant compared with wild-type, and some of these DEGs are involved in cellular metal ion homeostasis and transition metal ion homeostasis. Changes in expression of overlapping genes between the flr1 mutant and in plants under low-Ca2+ treatment were consistent in terms of direction, indicating that FLR1 is involved in Ca2+ homeostasis. In summary, we detected FLR1-mediated resistance to M. oryzae, a phenomenon associated with Ca2+ homeostasis.

Simultaneous quantification of fourteen characteristic active compounds in Eucommia ulmoides Oliver and its tea product by ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS).

Various kinds of bioactive compounds contribute to versatile health-promoting properties of Eucommia ulmoides Oliver (E. ulmoides). In present study, we developed a UPLC-QqQ-MS/MS method for simultaneous quantification of fourteen characteristic active compounds, including 3 lignans, 4 iridoids, 3 flavonoids and 4 phenolics in E. ulmoides and its tea product for the first time. The running time of the method is 6.5 min. It has good linearity, sensitivity, precision, accuracy, and stability. Using this high-throughput method, the distributions of fourteen characteristic active compounds in E. ulmoides and its tea product were clarified. Also, it was found that E. ulmoides tea exhibited superiority in contents of chlorogenic acid as compared with natural resources. Overall, the study provided a rapid, reliable, and efficient analysis method, which could be applied for the quality evaluation of E. ulmoides natural resources and their relative products in the field of food and medicine.

Effect of Bloodletting at Shaoshang and Shangyang Acupuncture Points on Outcome and Prognosis of Severe Community-Acquired Pneumonia in the Elderly.

OBJECTIVES: The aim of this study was to explore, whether treatment with bloodletting at Shaoshang and Shangyang acupuncture points would affect therapy outcome and prognosis for severe community-acquired pneumonia (SCAP) in the elderly. METHODS: A total of 62 patients, who met the diagnostic criteria for SCAP, were enrolled in the study and randomly divided into two groups, i.e., treatment group (n = 31) and control group (n = 31). All patients received a therapy according to the Chinese Clinical Practice and Expert Consensus of Emergency Severe Pneumonia from 2016. In addition to that, a bloodletting at Shaoshang (LU11) and Shangyang (LI1) acupuncture points was applied for the treatment group. This intervention was repeated for three times (ones daily), bloodletting a volume of 2-3 ml at each time point. Differences in a main index of clinical efficacy, body temperature (T), respiratory rate (RR), heart rate (Hr), white blood cell count (WBC), neutrophil percentage (N%), and C-reactive protein level (CRP) as well as different scores (CURB-65 score, SOFA score, and Apache II score) were compared between groups. Moreover, the 28-day mortality was compared between treatment and control group. The statistical methods involved in carrying out the current study include t-test, Wilcoxon test, and chi-square test. RESULTS: The clinical effective rate of the treatment group was 82.9%, which was significantly higher than the 17.1% in the control group (P < 0.05). After finishing the intervention, the treatment group showed significantly lower T (37.28 ± 0.54 vs. 37.82 ± 0.81), RR (20.06 ± 2.67 vs. 23.71 ± 6.85), Hr (81.71 ± 10.38 vs. 93.84 ± 15.39), CUBR-65 score (2.16 ± 0.74 vs. 3.03 ± 0.98), and SOFA score (5.84 ± 3.83 vs. 8.16 ± 4.2) compared to the control group (P < 0.05). The 28-day mortality rate of the treatment group was significantly lower than in the control group (12.9% vs. 45.2%, P = 0.05). CONCLUSIONS: Bloodletting at Shaoshang and Shangyang acupuncture points can support improving the clinical treatment efficacy for SCAP and reduce the 28-day mortality rate in the elderly.

Mutation at Different Sites of Metal Transporter Gene OsNramp5 Affects Cd Accumulation and Related Agronomic Traits in Rice (Oryza sativa L.).

OsNramp5 is a key gene involved in the control of the uptake of Cd, Mn, and other metal ions by rice root cells. The functional deficiency of this gene can significantly reduce the accumulation of Cd in rice grains, but the effects of its mutation on agronomic traits such as yield and quality have not been investigated comprehensively yet. In the present study, three Huanghuazhan-based OsNramp5 mutants [LCH1 (Low Cadmium Huanghuazhan 1), LCH2 (Low Cadmium Huanghuazhan 2), and LCH3 (Low Cadmium Huanghuazhan 3)] were obtained using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology. The mutation-type analysis showed that LCH1, LCH2, and LCH3 encoded defective OsNramp5 protein sequences containing only 76aa, 176aa, and 266aa, respectively. The determination of metal content and the statistics of related agronomic traits revealed that the functionally deficient OsNramp5 not only significantly reduced the accumulation of Cd in the grains of the mutants but also affected rice yield and quality. However, with the decrease of OsNramp5 mutation degree, its effects on chlorenchyma Mn accumulation, yield, and quality were also diminished. Additionally, we also found that the increase in the concentration of Mn in the soil restored the phenotype of the declined yield and quality due to the functional deficiency of OsNramp5. Our findings provide novel insights into and new materials for breeding rice varieties with low Cd accumulation and excellent agronomic traits under severe Cd pollution environment.

Effectiveness of Chinese Herbal Medicine Combined with Antibiotics for Extensively Drug-Resistant Enterobacteria and Nonfermentative Bacteria Infection: Real-Life Experience in a Retrospective Cohort.

Chinese herbal medicines (CHMs) have been successfully used in the treatment of infectious diseases, yet the effectiveness of CHMs for extensively drug-resistant enterobacteria (XDRE) infection remains unclear. Herein we developed a retrospective multicenter study including 766 patients with XDRE and nonfermentative bacteria (NFB) infection to investigate the effectiveness of CHMs combined with antibiotics in the treatment of XDRE infections in clinical daily practice in a cohort of patients and compared the regular antibiotics monotherapy. After 14-day treatment, the 547 patients accepted CHMs combined with antibiotics therapy indicating a more desirable effectiveness compared to the 219 patients treated with antibiotics monotherapy. The primary evaluation indexes included white blood cell count (WBC) and percentage of neutrophil (N%) in blood test. Secondary evaluation indexes consisted of body temperature, breath, heart rate, platelets, hemoglobin, red blood cell, albumin, creatinine, glucose, and 28-day survival rates. Briefly speaking, in our experience, CHMs combined with antibiotics therapy achieved more desirable effectiveness in treating XDRE infections compared with antibiotics monotherapy, and CHMs might be a potential huge resource in the field of XDRE infection management and enlighten the new antibiotics research and development. This trial is registered with ChiCTR-ORC-17011760.