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.

MiR-16 inhibits hepatocellular carcinoma progression by targeting FEAT through NF-κB signaling pathway.

OBJECTIVE: MicroRNA-16 (miR-16) expression has been proved to take part in the initiation and development of several cancers, including hepatocellular carcinoma (HCC). However, its role and its molecular mechanism in HCC cells remain unclear. Our study aimed to elucidate miR-16 probable role and potential mechanism in HCC cells. PATIENTS AND METHODS: MiR-16 expression in HCC was measured by Real Time-Polymerase Chain Reaction (RT-PCR). MiR-16 mimic or inhibitor was applied to increase or decrease miR-16 expression in Huh7 cells separately. The cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). The invaded cells and migrated cells were detected by the transwell assay. The epithelial-mesenchymal transition (EMT) and the nuclear factor-κB (NF-κB) were performed using Western blot. The tumor growth was measured via xenograft tumor formation assay. Moreover, bioinformatical methods and luciferase reporter assay were carried out to confirm the miR-16 target gene. RESULTS: MiR-16 expression was downregulated in HCC tissues and cells. Furthermore, the increasing miR-16 expression was suppressed, whereas the decreasing miR-16 expression promoted cell proliferation, invasion, and migration in Huh7 cells. Moreover, miR-16 targeted FEAT in regulating HCC progression. FEAT was associated with a poor prognosis of HCC patients. Especially, miR-16 suppressed EMT and NF-κB pathway in HCC and inhibited the tumor growth in vivo. CONCLUSIONS: We stated that miR-16 suppressed HCC cell progression by targeting FEAT and inhibiting EMT and NF-κB pathway. MiR-16 may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.

[Comparison of the NoSAS score with four different questionnaires as screening tools for obstructive sleep apnea-hypopnea syndrome].

Objective: To evaluate the clinical utility of the NoSAS score in the screening of patients with obstructive sleep apnea-hypopnea syndrome(OSAHS), and to compare the performance of the NoSAS score with other tools including Epworth Sleepiness Scale(ESS), STOP, STOP-Bang(SBQ) and Berlin questionnaires. Methods: A total of 444 consecutive patients(328 males and 116 females) with suspected OSAHS who underwent an overnight polysomnography(PSG) were recruited into this study. Five questionnaires including the NoSAS score, ESS, STOP, SBQ and Berlin were completed. Based on the severity of OSAHS which was determined by apnea-hypopnea index(AHI), the patients were classified into 4 groups: normal(<5 events/h), mild(5-15 events/h), moderate(15-30 events/h) and, severe (≥30 events/h) OSA.Sensitivity, specificity, positive predictive values, negative predictive values and the area under the receiver operating characteristics curve of 5 questionnaires were calculated. Results: With AHI≥5 events/h as the standard diagnosis of OSAHS, the NoSAS score and SBQ questionnaire showed a moderate performance, with the NoSAS score having the largest area under the ROC curve(0.753, P<0.001), followed by the SBQ questionnaire (0.727, P<0.001). The performance of the ESS, Berlin, and the STOP questionnaire was not high. Using mild moderate-severe(≥5 events/h), moderate-severe(≥15 events/h), and severe(≥30 events/h)OSAHS as cutoffs, NoSAS had the highest specificity and positive predictive values(80.2% and 88%, 72% and 69.8%, 66.3% and 50.5%), and the sensitivity and negative predictive values were (51.5% and 36.9%, 56.5% and 59.1%, 66.3% and 74.2%) .SBQ had the highest sensitivity and the negative predictive values(80.2% and 88%, 72% and 69.8%, 66.3% and 50.5%), and the specific and positive predictive values were (45.7% and 81.0%, 39.1% and 61.9%, 34.8% and 44.4%). The NoSAS score ≥ 7 had higher sensitivity and negative predictive value(75.0% and 47.1%, 78.1% and 66.5%, 82.7% and 81.9%)than the NoSAS socre ≥ 8. With AHI≥5 events/h as the standard diagnosis of OSAHS, the NoSAS score and the SBQ questionnaire had a higher accuracy than the other 3 questionnaires as screening questionnaires for diagnosing OSAHS, and the value of DOR were 4.298 and 3.758 respectively. Conclusions: The NoSAS score and the SBQ questionnaire have a moderate performance in diagnosing OSAHS. The NoSAS score is a new screening tool, and it is similar to the SBQ questionnaire, being also simple and effective. While the SBQ questionnaire is more widely used, it is necessary to further evaluate the diagnostic value of NoSAS score.

An Intensive Diabetes Screening and Treatment Program Improves Diagnosis, Treatment and Outcomes of Diabetes in Patients Admitted with Cardiac Diseases.

Aim: Patients with cardiac diseases, especially ischemic heart disease, are known to have a high prevalence of diabetes mellitus (DM). They are at risk of having inadequate glucose control. An intensive diabetes screening and treatment program was developed to identify and treat DM in patients admitted with cardiac diseases. Methods: Adult inpatients of 2 cardiac wards, namely Ward-A and Ward-B, at Nanjing Hospital, Nanjing, China, were studied. Patients were randomly assigned into either ward. In addition to routine examination and treatment, an intensive screening and treatment program to identify and treat patients with DM or impaired glucose regulation (IGR) was only applied in Ward-A patients. The glycated serum protein concentration, the length of hospitalization, and medical and total hospital cost were compared between the 2 wards. Results: The prevalence of DM was 17.85% in Ward-B. With implementation of this program, DM was higher in Ward-A (29.7%) and the prevalence of IRG was 7.8%. The overall prevalence of abnormal glucose metabolism was 37.5% in Ward-A. This program is associated with significantly reduced medical cost and length of inhospital days in patients requiring percutaneous coronary intervention (PCI) and reduced both the medical and total hospital costs in patients without PCI of Ward-A as compared with those of Ward-B who received standard treatment. Conclusion: The intensive screening and treatment program increases diagnosis rate of DM and IRG in inpatient with cardiac diseases, more effectively controls hyperglycemia, and is associated with shorter length of inhospital days and lower medical and total hospital costs. The trial registry number: ChiCTR-IPR-15007487.