Pancreatic cancer is a highly lethal malignancy with poor prognosis due to the lack of early symptoms and resultant late diagnosis. Thus, it is extremely urgent to establish a simple and effective method for the early diagnosis of pancreatic cancer. Although some studies have provided positive evidence for the use of exosomal surface protein glypican-1 (GPC1) as a biomarker for early screening, its clinical application is still controversial. Here, we systematically verified the role of exosomal GPC1 as a potential screening biomarker. First, bottleneck problems of a stable detection method and an identification standard were systematically studied, and a Python-based standardized data processing method was established to analyze exosomal GPC1 expression. Second, a detection panel consisting of exosomal GPC1, exosomal cluster of differentiation 82 (CD82), and serum carbohydrate antigen 19-9 (CA19-9) was employed for pancreatic cancer detection. This panel exhibited excellent diagnostic results (AUC = 0.942) and could effectively distinguish healthy people from patients with pancreatic cancer (P value threshold = 0.2282) and patients with pancreatitis from patients with pancreatic cancer (P value threshold = 0.5467). Implications: These results indicate that the combined detection of exosomal GPC1, exosomal CD82, and serum CA19-9 shows great promise as a standard method for pancreatic cancer detection and that this panel could be further applied for screening pancreatic cancer in Chinese populations.

Background: Alectinib significantly improves survival of non-small cell lung cancer (NSCLC) patients with anaplastic lymphoma kinase (ALK)-rearrangement. In this study, we analyzed the effects of different ALK rearrangements and co-mutations on the efficacy of alectinib. Methods: Using the electronic medical record system, we reviewed in terms of clinical and pathological features patients with advanced (IIIB/IV stage) ALK-rearranged NSCLC at Shanghai Chest Hospital between January 2018 and December 2021 who were treated with alectinib in first or second line and were assessed for objective response rate (ORR), disease control rate (DCR), and progression-free survival (PFS). Results: A total of 66 patients were enrolled in the study, and 17 types of ALK rearrangements were detected, of which five types of ALK rearrangements responded well to alectinib. We classified ALK-rearrangements into four main types, namely echinoderm microtubule-associated protein-like 4 (EML4)-ALK (E6:A20), EML4-ALK (E13:A20), EML4-ALK (E20:A20), and others. There was no significant difference in ORR and DCR of these types (ORR: 31.3% vs. 13.0% vs. 18.2% vs. 17.6%, P=0.575; DCR: 93.8% vs. 95.6% vs. 100.0% vs. 88.2%, P=0.627). The 3-year PFS rates were 25.0% (4/16) vs. 13.0% (3/23) vs. 27.3% (3/11) vs. 18.8% (3/16) for EML4-ALK (E6:A20), EML4-ALK (E13:A20), EML4-ALK (E20:A20), and others, respectively (P=0.725). The results of co-mutation analysis showed that the median PFS (mPFS) for patients with tumors harboring TP53 mutations was 30.4 months, significantly shorter than that of patients with tumors without co-mutations and whose mPFS was not mature (P=0.026). TSC1 co-mutation was also identified as a detrimental factor in outcome, with a DCR of 60% vs. 100% (P=0.031), mPFS of 30.4 months vs. not applicable (P=0.160) in patients with vs. those without this co-mutation, respectively. The efficacy of alectinib in patients with brain metastases is comparable to that in patients without distant organ metastases. There were two cases with specific fusion types that also responded to alectinib; namely, double ALK-rearrangements: EML4-ALK (E13:A20) and MSH2-ALK (M7:A20), and with a rare fusion partner, SPECC1L-ALK (S8:A20). Their PFS were 8.7 and 38.0 months, respectively. Conclusions: In this study, the efficacy of alectinib in different types of ALK-rearrangements varied slightly. TP53 and TSC1 co-mutations were identified as detrimental factors affecting efficacy. This study provides references for the response to alectinib in patients with different types of ALK rearrangements and co-mutation.

Background: Valid cancer screening and treatment monitoring are critical for cancer patients. Although genitourinary system cancers have a high recurrence rate, when diagnosed, patients undergo surgery promptly. We sought to explore sensitive and noninvasive screening and postoperative recurrence monitoring methods, such as circulating tumor cells (CTCs) or tumor susceptibility gene detection, to determine their appropriateness for genitourinary system cancers. Methods: We adopted multiple detection methods. Enrichment-immunofluorescence in situ hybridization (SE-iFISH) was employed to detect CTCs from the peripheral blood of patients, and Agena Bioscience MassARRAY was used to detect single-nucleotide polymorphisms (SNPs) in tumor susceptibility genes from urine cells. Results: In our research, CTCs showed a 76.92% positivity rate among 26 genitourinary system cancer patients, and the number of CTCs was consistent with the stage of cancer. In monitoring for bladder cancer (BC) recurrence, CTCs were more prevalent than urine cytology (66.67% vs. 41.67%). To our surprise, urine cellular XPC (rs2228001, A2815C) and XRCC1 (rs25487, G1196A) polymorphisms were specifically found in cancer patients but not in patients with inflammation or in healthy individuals. XPC polymorphism (rs2228001, A2815C) rates were 30.77%, 40%, and 50% in bladder cancer, renal carcinoma, and prostate cancer patients, respectively, and those for XRCC1 (rs25487, G1196A) were 3.85%, 20%, and 25%, respectively. Conclusions: As a common biomarker, CTCs showed remarkable performance in cancer screening and monitoring. The noninvasive panel comprising CTCs and XPC (rs2228001, A2815C) and XRCC1 (rs25487, G1196A) polymorphisms showed high sensitivity (positive rate: 92.86%) and is suitable for genitourinary system cancer screening.

Background: Previous studies have suggested associations between testosterone, genetic factors, and a series of complex diseases, but the associations with the lifespan phenotype, such as health span, remain unclear.Methods: In this prospective cohort study, we analyzed 145,481 men and 147,733 women aged 38–73 years old from UK Biobank (UKB) to investigate the sex-specific associations of total testosterone (TT), free testosterone (FT), or polygenic risk score (PRS) with health span termination (HST) risk. At baseline, serum testosterone levels were measured. HST was defined by eight events strongly associated with longevity. PRS, an efficient tool combining the effect of common genetic variants to discriminate genetic risk of complex phenotypes, was constructed by 12 single-nucleotide polymorphisms related to health span from UKB (P ≤ 5.0 × 10⁻⁸). We used multivariable Cox regression models to estimate hazard ratios (HRs) and 95% confidence intervals (CIs).Results: With a median follow-up time of 7.70 years, 26,748 (18.39%) men and 18,963 (12.84%) women had HST. TT was negatively associated with HST in men [HR per standard deviation (SD) increment of log-TT: 0.92, 95% CI: 0.88–0.97]. Inversely, both TT (HR per SD increment of log-TT: 1.05, 95% CI: 1.02–1.08) and FT (HR per SD increment of log-FT: 1.08, 95% CI: 1.05–1.11) presented an increased risk of HST in women. PRS was positively associated with HST risk (quintile 5 versus quintile 1, men, HR: 1.19, 95% CI: 1.15–1.24; women, HR: 1.21, 95% CI: 1.16–1.27). Moreover, men with high TT and low genetic risk showed the lowest HST risk (HR: 0.80, 95% CI: 0.73–0.88), whereas HST risk for women with both high TT and genetic risk increased obviously (HR: 1.32, 95% CI: 1.19–1.46). Similar joint effects were observed for FT in both genders.Conclusions: We observed sex-specific associations that testosterone was negatively associated with HST risk in men and positively associated with HST risk in women. Genetic factors increased the HST risk, suggesting that participants with both high genetic risk and abnormal testosterone levels (high level in women or low level in men) should be the target for early intervention. Although our findings highlight the associations between testosterone and health span, further mechanistic studies and prospective trials are warranted to explore the causation behind.

Background. Circulating tumor-derived endothelial cell (CTEC) is a new potential tumor biomarker to be associated with cancer development and treatment efficacy. However, few evidences are available for breast cancer. Methods. Eighty-nine breast cancer patients were recruited, and preoperative and postoperative blood samples were collected. Besides, 20 noncancer persons were enrolled as controls. An improved subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH) method was adopted to codetect CD31+ aneuploid CTEC and CD31− aneuploid circulating tumor cell (CTC). Then, the clinical significance of CTCs and CTECs on breast cancer screening and prognosis prediction was evaluated and compared. Results. The positive rate of CTCs and CTECs in newly diagnosed breast cancer patients was 68.75% and 71.88%. Among detected aneuploid circulating rare cells, CTEC accounts for a greater proportion than CTC in breast cancer patients. CTEC-positive rate and level were significantly higher in breast cancer patients with lymph node metastasis (LNM) than those without LNM ( ), while there was no significant difference in CTC. CTEC (area under the curve, AUC = 0.859) had better performance than CTC (AUC = 0.795) to distinguish breast cancer patients from controls by receiver operator characteristic curve analysis. Preoperative CTEC count ≥ 2 was a significant risk factor for reducing PFS of breast cancer patients. Conclusions. CTECs may function as a reliable supplementary biomarker in breast cancer screening and prognosis prediction.

Circulating tumor cells (CTCs) are widely used in cancer screening and monitoring. The present study focused on investigating the optimal time for the postoperative CTC detection in patients with colorectal cancer (CRC) to obtain more accurate results and facilitate subsequent treatment. By subtraction enrichment immunofluorescence in situ hybridization detection of CTCs, the present study demonstrated that different postoperative detection times in CRC substantially influenced the CTC numbers. In total, 134 subjects were enrolled. Among 10 healthy individuals and 20 preoperative patients with CRC, no CTCs were identified in the healthy subjects, and CTCs were detected in 85% (17/20) of the preoperative patients. In total, 104 postoperative patients with CRC (53 males and 51 females) with a mean average age of 57.63 years were studied. The total CTC detection rate was 81.73% (85/104) and the mean average CTC numbers in patients with tumor stage (T) T1, T2, T3 and T4 were 4.00, 3.33, 5.90 and 5.64 per 7.50 ml of peripheral blood, respectively. The CTC number trends in these four tumor stages within 5, 6, 7 and 10 postoperative days were variable, and were the most stable at 7 days. Gradual upward trends in CTC numbers were observed after 5, 6 and 7 postoperative days, and this upward trend was more obvious after 7 days. Overall, the findings of the present study suggest that CTC detection in CRC should be performed after at least 7 postoperative days rather than within 7 postoperative days.