Unintrusive multi-cancer detection by circulating cell-free DNA methylation sequencing (THUNDER): development and independent validation studies.

BACKGROUND: Early detection of cancer offers the opportunity to identify candidates when curative treatments are achievable. The THUNDER study (THe UNintrusive Detection of EaRly-stage cancers, NCT04820868) aimed to evaluate the performance of enhanced linear-splinter amplification sequencing, a previously described cell-free DNA (cfDNA) methylation-based technology, in the early detection and localization of six types of cancers in the colorectum, esophagus, liver, lung, ovary, and pancreas. PATIENTS AND METHODS: A customized panel of 161 984 CpG sites was constructed and validated by public and in-house (cancer: n = 249; non-cancer: n = 288) methylome data, respectively. The cfDNA samples from 1693 participants (cancer: n = 735; non-cancer: n = 958) were retrospectively collected to train and validate two multi-cancer detection blood test (MCDBT-1/2) models for different clinical scenarios. The models were validated on a prospective and independent cohort of age-matched 1010 participants (cancer: n = 505; non-cancer: n = 505). Simulation using the cancer incidence in China was applied to infer stage shift and survival benefits to demonstrate the potential utility of the models in the real world. RESULTS: MCDBT-1 yielded a sensitivity of 69.1% (64.8%-73.3%), a specificity of 98.9% (97.6%-99.7%), and tissue origin accuracy of 83.2% (78.7%-87.1%) in the independent validation set. For early-stage (I-III) patients, the sensitivity of MCDBT-1 was 59.8% (54.4%-65.0%). In the real-world simulation, MCDBT-1 achieved a sensitivity of 70.6% in detecting the six cancers, thus decreasing late-stage incidence by 38.7%-46.4%, and increasing 5-year survival rate by 33.1%-40.4%, respectively. In parallel, MCDBT-2 was generated at a slightly low specificity of 95.1% (92.8%-96.9%) but a higher sensitivity of 75.1% (71.9%-79.8%) than MCDBT-1 for populations at relatively high risk of cancers, and also had ideal performance. CONCLUSION: In this large-scale clinical validation study, MCDBT-1/2 models showed high sensitivity, specificity, and accuracy of predicted origin in detecting six types of cancers.

[Clinical value of transcatheter arterial chemoembolization therapy for hepatocellular carcinoma with blood supply from right adrenal artery].

Objective: To study the clinical value of transcatheter arterial chemoembolization (TACE) therapy for hepatocellular carcinoma with blood supply from right adrenal artery. Methods: An imaging and clinical data of HCC patients with blood supply from right adrenal artery were collected from 2012 to 2016 after TACE treatment in the Second Affiliated Hospital of Chongqing Medical University and the safety and therapeutic efficacy of complete embolization therapy was analyzed retrospectively. Results: Twenty hepatocellular carcinoma patients with blood supply from right adrenal artery had received 23 times treatment. All lesions had invaded and protruded from the exogenous development of liver capsule. There were 14 cases with lesions > 5 cm in diameter. Right adrenal artery was found to be involved in the blood supply of three cases of hepatocellular carcinoma during TACE treatment for the first time. In addition, the remaining 17 cases had also received TACE treatment for the second to sixth time. The superior, middle, and inferior adrenal arteries were involved in 13, 3, and 9 cases, respectively. Twenty-four right adrenal arteries (96.0%) superselectively cannulated were successfully embolized without any serious complications. The standard method for evaluating the efficacy of liver cancer in 20 solid tumors follow-up cases showed that three cases were completely relieved, nine cases were partially relieved, two cases were stable, and six cases were progressive. The effective rate of embolization with blood supply from right adrenal artery lesions was 60.0%, and the control rate of lesion development was 70.0%. Conclusion: The right adrenal artery is mainly located in the S5-S7 segments of the liver. TACE features are obvious to ascertain its safety and effectiveness in the treatment of right adrenal artery tumors.

First Report of Root Rot on Atractylodes macrocephala (Largehead Atractylodes Rhizome) Caused by Ceratobasidium sp. in China.

Atractylodes macrocephala is a perennial herbaceous plant (family Asteraceae) native to China. The biennial root, Largehead Atractylodes Rhizome (LAR), is the most commonly used Chinese herbal medicine to prevent early pregnancy loss due to miscarriage. From summer 2010 to spring 2012, symptoms of root rot were observed on LAR in Xianfeng county, Enshi city, Hubei Province, China. White mold on the root of LAR could be observed at an early growth stage in the field and the white mold spread over the entire plant after 10 days, which differs from root rot of LAR caused by Fusarium oxysporum and Rhizoctonia solani, neither of which are characterized as having mycelium spreading over the whole plant (4). Where root rot symptoms were present, rhizome yield was reduced by 15% on average, with up to 40% yield loss in some fields. Under humid conditions in mid-June, the disease in the field spread quickly and the rhizomes of LAR were completely rotted. After rainfall and increasing temperature from 16 to 35°C, white mycelium appeared and plants withered within a few weeks. In April 2011 and 2012, a fungus was consistently recovered from symptomatic rhizome samples after they were surface sterilized with 0.1% mercuric chloride solution and plated onto potato dextrose agar (PDA). Pale gray colonies with short aerial mycelia and brown sclerotia formed on PDA after 7 days incubation at 28°C. Binucleate cells were observed using light microscopy and the characteristics were matched with morphological characteristics of a Ceratobasidium sp (3). Genomic DNA of the culture was extracted, and the rDNA-internal transcribed spacer sequence (GenBank Accession No. JQ926741) showed 99% identity to Ceratobasidium sp (GenBank No. H269825.1). Mycelial plugs of the culture taken from PDA were inoculated onto 40 rhizomes of 1-year-old seedlings and plants were incubated with a 16-h photoperiod at 28°C and 90% relative humidity in an artificial climate chamber where they developed typical disease symptoms after 2 days. Ten rhizomes of 1-year-old seedlings and were treated with PDA plugs only. All seedlings inoculated with the pathogen were withered and the rhizomes were completely covered with gray mycelium 2 days after inoculation, which was similar to the symptoms observed in the field. After 7 days, the symptoms were more severe than those observed in the field, with seedlings rotted completely. The main stalk of all inoculated plants was covered with gray mycelia in 4 days, and the stalk became withered, which was similar to the symptoms observed in the field. No symptoms were observed on control seedlings and plants. Koch's postulates were fulfilled by successful reisolation of Ceratobasidium sp. from diseased seedlings. The pathogenicity tests were carried out twice. Ceratobasidium sp. has been reported to cause root rot of canola in Washington (2). It has also been observed on Rehmannia in China (1). To our knowledge, this is the first report of Ceratobasidium sp. causing root rot on LAR. References: (1) B. B. Chen et al. Chin. J. Chin. Material Medica (In Chinese) 9:1137, 2011. (2) K. L. Schroeder et al. Plant Dis. 96:591, 2012. (3) B. Sneh et al. Page 39 in: Identification of Rhizoctonia Species. The American Phytopathological Society, 1991. (4) S. X. Zang et al. J. Agric. Univ. Hebei (In Chinese) 28:73, 2005.

[Epidemiological trend of early-onset gastric cancer and late-onset gastric cancer in China from 2000 to 2019].

Objective: In order to understand the changing trends of gastric cancer incidence and mortality in early-onset and late-onset in China from 2000 to 2019. Methods: The Global Burden of Disease research data was collected, and Excel and R 4.2.1 softwares were used to examine the incidence rate, mortality rate, and disability-adjusted life years (DALY) of Chinese people from 2000 to 2019, with a focus on gender, age, and year. Results: In 2019, the crude incidence rates were 7.06/100 000 (95%UI: 6.63/100 000-7.59/100 000) and 114.52/100 000 (95%UI: 108.79/100 000-121.63/100 000) for early- and late-onset gastric cancer, respectively. The crude mortality rate for early-onset gastric cancer was 3.29/100 000 (95%UI: 3.11/100 000- 3.50/100 000), while the crude mortality rate for late-onset gastric cancer was 81.88/100 000 (95%UI: 78.15/100 000-86.04/100 000). Additionally, the crude DALY rates for these two types of gastric cancer were 156.48/100 000 (95%UI: 148.82/100 000-165.84/100 000) and 1 750.13/100 000 (95%UI: 1 661.21/100 000-1 852.99/100 000). The standardized incidence of early-onset gastric cancer decreased from 5.49/100 000 in 2000 to 4.76/100 000 in 2019, and that of late-onset gastric cancer decreased from 143.45/100 000 in 2000 to 123.02/100 000 in 2019.The standardized mortality rate of early-onset gastric cancer decreased from 4.16/100 000 in 2000 to 2.18/100 000 in 2019, and that of late-onset gastric cancer decreased from 140.82/100 000 in 2000 to 91.49/100 000 in 2019. The standardized DALY rate for early-onset gastric cancer in 2019 was 105.87/100 000 (95%UI: 87.98/100 000 -125.60/100 000), lower than 198.84/100 000 (95%UI: 179.47/100 000- 219.83/100 000) in 2000. The standardized DALY rate for late onset gastric cancer in 2019 was 1 821.11/100 000 (95%UI: 1 509.42/100 000-2 158.53/100 000), lower than 2 932.52/100 000 (95%UI: 2 665.92/100 000-3 252.60/100 000) in 2000. Conclusions: The standardized mortality rate of early-onset gastric cancer in China showed a decreasing trend from 2000 to 2019. The standardized mortality rate of late onset gastric cancer showed a trend of first increasing and then decreasing. Notably, the incidence, mortality, and DALY of late-onset gastric cancer were significantly higher than those of early-onset gastric cancer during this period. Additionally, male incidence, mortality, and crude DALY rates were higher than female.

Physiological dissection revealed that both uptake and assimilation are the major components regulating different growth responses of two tobacco cultivars to nitrogen nutrition.

K326 and HD represent major tobacco cultivars in China, which required large N fertiliser input but at different application rates. To understand primary components affecting tobacco N use physiology, we adopted these two varieties as valuable genetic material to assess their growth response to N nutrition. We established a hydroponic culture system to grow plants supplied with different N regimes. Plant biomass, N, ammonium, nitrate, arginine, GS and NR activity, N transfer and use efficiency as well as root uptake were examined. Our data revealed the preference of K326 and HD to utilise nitrate or ammonium nitrate but not ammonium alone, with 2 mm N supply probably sufficient and economical to achieve good biomass production at the vegetative stage. Moreover, both varieties were very sensitive to ammonium, perhaps due to lack of or abnormal signalling related to nitrate and/or arginine rather than impairment of N acquisition and initial assimilation; this was supported by measurements of the plant content of N, ammonium and activities of GS and NR. Notably, short-term 15 N root influx studies identified differential uptake kinetics of K326 and HD, with distinct affinities and transport rates for ammonium and nitrate. The data suggest that the growth adaptation of K326 or HD to higher or lower N may be ascribed to different competences for effective N uptake/translocation and assimilation. Thus, our work provides valuable information to prompt deeper investigation of the molecular basis controlling plant N use efficiency.