[Clinicopathological features and prognosis of early-onset gastric cancer: a large-scale retrospective real-world study].

Objective: To clarify the clinicopathological features, prognosis, and recurrence pattern of early-onset gastric cancer (EOGC). Methods: Using data from the gastric cancer database of Zhongshan Hospital, Fudan University, we performed a retrospective, large-scale, real-world study of 5046 patients with gastric cancer who had undergone redical or palliative gastrectomy from January 2013 to December 2018, including 425 patients with EOGC (age ≤45 years) and 4621 controls. All those patients were pathologically confirmed adenocarcinoma with complete follow-up of five years. Residue gastric cancer and patients without complete clinical or follow-up data were excluded. We used a combination of outpatient and telephone follow-up, ending in October 2022 (median duration of follow-up 60 months), and compared the clinicopathological features and prognosis of the two groups. Results: The clinicopathological features of EOGC included female predominance (61.1% [262/425 vs. 26.3% [1217/4621], χ2=234.215, P<0.001), fewer comorbidities (31.3% [133/425] vs. 58.5% [2703/4621], χ2=34.378, P<0.001), poorer differentiation (90.6% [385/425] vs. 78.2% [3614/4621], χ2=30.642, P<0.001), higher proportion of diffuse type (53.9% [229/425] vs. 18.3% [846/4621], χ2=274.474, P<0.001), higher proportion of T4 stage (44.7% [190/425] vs. 37.5% [1733/4621], χ2=17.535, P=0.001), more lymph node metastases (60.5% [257/425] vs. 53.9% [2491/4621], χ2=6.764, P=0.009), and higher proportion of pathological stage III/IV (47.5% [202/425] vs. 42.4% [1959/4621], χ2=4.093, P=0.043). The 5-year overall survival rates of the EOGC and control groups were 55.1% and 49.1%, respectively. Overall survival was significantly better in the EOGC than in the control group (P<0.001). According to subgroup analysis, the prognosis of pathological stage I/II/III EOGC was better than that of the control group. Recurrence rates were similar in the two groups, whereas patients with EOGC had a higher proportion of peritoneal recurrence (7.8% [33/425] vs. 3.2% [146/4621], χ2=23.741, P<0.001) and a lower proportion of distant metastasis (4.9% [21/425] vs. 8.3% [385/4621], χ2=6.247, P=0.012). Conclusion: EOGC has unique clinicopathological features and recurrence patterns and resectable EOGC has a better prognosis, suggesting that patients with EOGC should be actively treated with the focus on preventing peritoneal recurrence.

[Value of autocrine motility factors in the prediction of the disease progression of PBC- associated hepatocellular carcinoma].

Objective: To clarify the value of autocrine motility factor (ATX) in predicting the disease progression of primary biliary cholangitis (PBC)-associated hepatocellular carcinoma (HCC). Methods: A prospective cohort of 179 newly diagnosed autoimmune liver disease (PBC) patients admitted to the Department of Hepatology at the Fifth Medical Center of the People's Liberation Army General Hospital from January 2016 to January 2018 was selected. All PBC patients received ursodeoxycholic acid (UDCA) treatment and were followed up.The endpoint of the follow-up was the occurrence of primary liver cancer. The relationship between ATX and the clinical characteristics of patients and its significance in predicting disease progression and HCC were analyzed. Results: The peripheral blood ATX level was significantly higher in PBC patients than that of alcoholic cirrhosis (t = 3.278, P = 0.001) and healthy controls (t = 6.594, P < 0.001), but there was no significant difference in ATX levels compared with patients with non-PBC- associated HCC (t = -0.240, P = 0.811). The expression of ATX in liver tissue of PBC patients was significantly higher than that of healthy individuals (Z = -3.633, P < 0.001) and patients with alcoholic cirrhosis (Z = -3.283, P < 0.001), while the expression of ATX in the advanced stage was significantly higher than that in early-stage PBC patients (Z = -2.018, P = 0.034). There was a significant difference in baseline ATX levels between PBC patients without HCC and PBC patients with HCC (228.451 ± 124.093 ng/ml vs. 301.583 ± 100.512 ng/ml, t = 2.339, P = 0.021). Multivariate logistic regression analysis showed that ATX was an independent predictor of PBC progression to HCC (OR = 1.245, 95%CI 1.097-1.413). The baseline peripheral blood ATX level in predicting AUROC of PBC-associated HCC was 0.714, 95%CI 0.597-0.857 and the sensitivity and specificity were 84.6%, and 59.0%, respectively. The optimal cutoff value for predicting serum ATX levels in the occurrence of HCC was 235.254 ng/ml. Conclusion: Patients with PBC have significantly higher levels of ATX expression in their peripheral blood and liver tissue, which can be utilized to assess treatment effectiveness and predict disease progression.

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.

Peroxiredoxin1 promotes cell proliferation, migration and invasion of colorectal cancer via p38MAPK signaling.

OBJECTIVE: Peroxiredoxin1 (PRDX1), a class of thiol peroxidases, is a multifunctional protein. We aimed at analyzing the effect of PRDX1 on proliferation, apoptosis, migration and invasion of colorectal cancer and to investigate the potential mechanism. MATERIALS AND METHODS: Western blot and PCR were used to validate the silencing efficiency in SW480 cell by transfection of PRDX1-siRNA. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) test. Flow cytometry Annexin V/PI double staining was used to analyze cell apoptosis. Transwell and scratch test were used to detect the migration and invasion of cells. Signal pathway protein expression was analyzed by Western blot. RESULTS: The expression of PRDX1 in SW480 cells could be reduced by siRNA effectively. The cell proliferation, migration and invasion were reduced significantly compared with control group after down-regulation of PRDX1 (p<0.05), while the cell apoptosis was enhanced significantly (p<0.05). The ratio of phospho-p38 mitogen-activated protein kinases (p-p38) /p38 mitogen-activated protein kinases (p38) was down-regulated after the down-regulation of PRDX1 (p<0.05). The ratio of phospho-c-Jun N-terminal protein kinase (p-JNK)/c-Jun N-terminal protein kinase (JNK) and phospho-extracellular regulated protein kinases (p-ERK)/extracellular regulated protein kinases (ERK) showed changes with no significant difference (p>0.05). CONCLUSIONS: Down-regulation of PRDX1 in colorectal cancer SW480 cells could inhibit the cell proliferation, migration, invasion, and induce cell apoptosis. This is very likely to be achieved by activating the p38MAPK-signaling pathway.

Expression of the fetal hematopoiesis regulator FEV indicates leukemias of prenatal origin.

The origin of cancers is associated with etiology as well as therapeutics. Several studies reveal that malignancies in children can originate in utero. However, a diagnostic approach to distinguish between cancers initiated pre- or postnatally is absent. Here we identified a transcriptional factor FEV (fifth Ewing variant) that was expressed in fetal hematopoietic cells and became silent after birth. We characterized that FEV was essential for the self-renewal of hematopoietic stem cells (HSCs). We next found that FEV was expressed in most infant leukemia samples, but seldom in adult samples, in accord with the known prenatal origins of the former. We further determined the majority of pediatric acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML) were FEV positive. Moreover, FEV knockdown markedly impaired the leukemia-propagating ability of leukemic stem cells. We therefore identified FEV is unique to fetal HSCs and stably expressed in leukemic cells of prenatal origin. It may also provide a tractable therapeutic target.

Reduced miRNA-218 expression in pancreatic cancer patients as a predictor of poor prognosis.

microRNA-218 (miR-218) is a vertebrate-specific miRNA that plays a crucial role in tumorigenesis and tumor progression. This study analyzed the miR-218 expression level and clinical significance in pancreatic cancer. One hundred and seven pairs of pancreatic cancer and adjacent normal tissues were analyzed by quantitative reverse-transcriptase polymerase chain reaction. The correlation between miR-218 expression and clinicopathological characters was determined by the two-sample Student t-test. The survival correlations were analyzed by the Kaplan-Meier method and Cox proportional hazards model. The relative expression of miR-218 in pancreatic cancer tissues (2.63 ± 1.59) was significantly lower than that in matched noncancerous pancreatic tissues (6.52 ± 2.50, P < 0.001). The low expression of miR-218 in the pancreatic cancer tissues were strongly correlated with the TNM classification (P = 0.02), distant metastasis (P = 0.001), and tumor differentiation (P = 0.003). The low level of miR-218 expression was significantly correlated with the shorter overall survival time of pancreatic cancer patients (5-year overall survival rate: 7.5 vs 34.9%; log-rank test: P < 0.001). Multivariate analyses confirmed that a low level of miR-218 expression was an independent predictor of poor prognosis in pancreatic cancer patients (Hazard ratio: 7.24; 95% confidence interval: 2.01-18.28; P = 0.007). Our findings suggested a significant downregulation in the expression of miR-218; this might have considerable potential value in the prognosis for pancreatic cancer.

MicroRNA-141 inhibits tumor growth and metastasis in gastric cancer by directly targeting transcriptional co-activator with PDZ-binding motif, TAZ.

Gastric cancer (GC) is a biologically heterogeneous disease accompanying various genetic and epigenetic alterations, and the molecular mechanisms underlying this disease are complex and not completely understood. Increasing evidence shows that abnormal microRNA (miRNA) expression is involved in GC tumorigenesis, but the role of specific miRNAs involved in this disease remains elusive. MiR-141 was previously reported to act as tumor suppressors or oncogenes in diverse cancers. However, their accurate expression, function and mechanism in GC are largely unclear. Here we found that the expression of miR-141 was significantly reduced in GC compared with paired adjacent normal tissues and was significantly correlated with a more aggressive phenotype of GC in patients. Ectopic expression of miR-141 mimics in GC cell lines resulted in reduced proliferation, invasion and migration, and inhibition of miR-141 in GC cell lines promoted cell proliferation, invasion and migration in vitro. We further demonstrated that miR-141 acted as tumor suppressors through targeting transcriptional co-activator with PDZ-binding motif (TAZ) in GC. Moreover, the inverse relationship between miR-141 and its target was verified in patients and xenograft mice. Finally, overexpression of miR-141 suppressed tumor growth and pulmonary metastasis in nude mice. Take together, we identified that miR-141 is a potent tumor suppressor in the stomach, and its growth inhibitory effects are, in part, mediated through its downstream target gene, TAZ. These findings implied that miR-141 might be employed as novel prognostic markers and therapeutic targets of GC.

MicroRNA-25 promotes gastric cancer migration, invasion and proliferation by directly targeting transducer of ERBB2, 1 and correlates with poor survival.

Gastric cancer (GC) is one of the most common tumors and the molecular mechanism underlying its metastasis is still largely unclear. Here, we show that miR-25 was overexpressed in plasma and primary tumor tissues of GC patients with tumor node metastasis stage (III or IV) or lymph node metastasis. MiR-25 inhibition significantly decreased the metastasis, invasion and proliferation of GC cells in vitro, and reduced their capacity to develop distal pulmonary metastases and peritoneal dissemination in vivo. Furthermore, miR-25 repressed transducer of ERBB2, 1 (TOB1) expression by directly binding to TOB1-3'-UTR, and the inverse correlation was observed between the expressions of miR-25 and TOB1 mRNA in primary GC tissues. Moreover, the loss of TOB1 increased the metastasis, invasion and proliferation of GC cells, and the restoration of TOB1 led to suppressed metastasis, invasion and proliferation. The receiver operating characteristics analysis yielded an area under the curve value of 0.7325 in distinguishing the GC patients with death from those with survival. The analysis of optimal cutoff value revealed poor survival in GC patients with high plasma concentrations of miR-25 (>0.2333 amol/µl). Taken together, miR-25 promotes GC progression by directly downregulating TOB1 expression, and may be a noninvasive biomarker for the prognosis of GC patients.

Postoperative chemoradiotherapy improves survival in esophageal squamous cell cancer with extracapsular lymph node extension.

In esophageal squamous cell carcinoma (ESCC), extracapsular extention (ECE) in metastatic lymph nodes portends high rate of recurrence and poor prognosis. To our knowledge, the effectiveness of postoperative chemoradiotherapy (CRT) in these patients has never been investigated. In this retrospective study, we compared the outcomes of surgery with or without postoperative chemoradiotherapy in ESCC patients with ECE. From 2008 to 2009, 90 ECSS patients with ECE were included. Among those patients, 47 only received curative surgery alone, and 43 received additional postoperative concurrent CRT which consisted of radiotherapy (median dose 50 Gy) and chemotherapy (5-fluorouracil 1000 mg/m(2), days 1-4 and 29-32; cisplatinum 25 mg/m(2), days 1-3 and 29-31). Patients treated with postoperative CRT had significantly more T3/4 tumors (p=0.023). Based on log-rank stratified by T stage, postoperative adjuvant CRT significantly improved the overall survival (p=0.017) and progression free survival (p=0.002). In multivariate analysis, adjuvant CRT was identified as an independent prognostic factor (HR=0.494, CI 0.290-0.844, p=0.010). Compared with surgery alone, the CRT group had significantly fewer cases of regional recurrence (P=0.048) and overall recurrence (P=0.024). However, there was no significant difference in distant metastasis between two groups (P=0.755). In conclusion, our data suggest that the postoperative adjuvant CRT might be beneficial in selected subgroups of ESCC patients with ECE. To further verify these results, a prospective trial with a large sample size is needed.

Pathological analysis of extracapsular extension of metastatic lymph node and its potential impact on nodal clinical target volume in the radiotherapy of esophageal squamous cell carcinoma.

There is no consensus regarding the clinical target volume (CTV) margins which surround the gross tumor volume of metastatic lymph nodes (LN) in radiotherapy of esophageal squamous cell carcinoma (ESCC). This study retrospectively assessed the distance of extracapsular extension (ECE) of metastatic LN in thoracic ESCC and defined nodal CTV margins. Histological sections of metastatic LNs from 217 patients with thoracic ESCC were re-examined. The incidence and maximal distance of ECE of metastatic LNs were assessed. The relationships between ECE and clinicopathologic features were also investigated. The ECE was found in 37.3% of patients (81/217) and 23.1% of metastatic LN (159/689), and the incidences had a significant relationship with N stage and LN size. The median distance of ECE was 1.0 mm (range, 0.2-9.7 mm). The distance of ECE showed a positive correlation with LN size (Spearman's correlation coefficient = 0.419; p<0.001). The ECE distances of LN with <10 mm diameter were significantly smaller than LN with 10-30 mm diameter (p<0.001). The 95th percentiles of ECE distances for these two groups were 3 mm and 5 mm, respectively. For pathologic LN <10 mm in diameter, a 3-mm CTV margin appears to be adequate to encompass 95% of the microscopic ECE, and for LN 10-30 mm, a 5-mm CTV margin is recommended.

The expression of glutathione peroxidase-1 and the anabolism of collagen regulation pathway transforming growth factor-beta1-connective tissue growth factor in women with uterine prolapse and the clinic significance.

OBJECTIVES: To investigate the expression of the anabolism of collagen regulation pathways connective tissue growth factor (CTGF) -transforming growth factor-betal (TGF-beta1) and glutathione peroxidase-1 (GPx1) in women with uterine prolapse and a study of the clinic significance. MATERIALS AND METHODS: The expression of TGF-beta1, CTGF, and GPx1 was detected by immunohistochemical staining in pubocervical fascia tissue of 30 women with uterine prolapse, including ten cases of POP-QII, ten cases of POP-QIII, ten cases of POP-QIV, and 20 cases were control group with non-prolapse and non-malignant lesions. RESULTS: There was a negative correlation between the POP-Q and expression of TGF-beta1. With the increase of POP-Q degree, the expression degree of TGF-beta1 decreased correspondingly, which also applied to CTGF and GPx1. On the other hand, there was a positive correlation between TGF-beta1 and CTGF. The synergistic change trend was found between TGF-beta1 and CTGF. It could also be seen between CTGF and GPx1 and betweenTGF-beta1 and GPx1. CONCLUSION: The expression of the antioxidase GPx1 in pelvic support structure of POP women was decreased, which resulted in the antioxidation reduced. It could break the balance of oxidation and antioxidation in pelvic support structure, and may induce an increase of ROS level and the down-regulation of TGF-beta1-CTGF pathway. It could inhibit the anabolism of collagen and injury the pelvic support structure, thus promoting the occurrence and development of POP.

Single nucleotide polymorphisms in NOS2A and NOS3 genes are not associated with treatment response of non-small cell lung cancer patients following the definitive radiochemotherapy.

Nitric oxide (NO), is endogenously synthesized from L-arginine by nitric oxide synthase (NOS), exhibits a dual role in sensitivity to radiotherapy and chemotherapy of cancer cells. The aim of this study was to evaluate the influence of polymorphisms in NOS genes on treatment response of non-small-cell lung cancer (NSCLC) patients after radiochemotherapy. A cohort of 198 NSCLC patients treated with radiochemotherapy between 2009 and 2011 were included in this study. Genotyping analyses of 35 SNPs ( NOS2A, 21 and NOS3, 14) in each sample were conducted by using the Sequenom MassArray system. Unconditional logistic regression was performed to assess the association between treatment response and each genotype while adjusting or not for other covariates. Of 198 patients, 87 (43.9%) had objective responses, and 111(56.1%) did not respond. We observed no significant associations between treatment response and each genotype. While adjusting for other covariates, the associations were also not significant. Our results suggest that genetic variations within the NOS2A and NOS3 genes may not influence the treatment response in NSCLC patients with radiochemotherapy. Future studies in this problem are required to confirm our findings.

Toll-like receptor 4 signaling promotes the immunosuppressive cytokine production of human cervical cancer.

OBJECTIVES: To investigate the expression of TLR-4 (toll-like receptor) on human cervical cancer and find the biological function of the TLR-4 signal system. METHODS: The immunohistochemistry method was performed to study the protein expression and distribution of TLR-4. The viability of HeLa cells was determined by cell viability assay. Cell proliferation was detected by FCM, ELISA and Western blot were used to observe the gene and protein expression of IL-6 and TGF-beta1 in Hela cell lines. RESULTS: TLR-4 was over-expressed in cervix cancer, and its activation by LPS promotes proliferation and anti-apoptosis in Hela cells in vitro. Moreover the cell line proliferation increased in a dose- and time-dependent manner. The production of IL-6 and TGF-beta1 were promoted through the activation of the NF-kappaB signaling pathway.

Survey of intestinal parasites in pigs from intensive farms in Guangdong Province, People's Republic of China.

The prevalence of intestinal parasites was investigated in intensive pig farms in Guangdong Province, China between July 2000 and July 2002. Faecal samples from 3636 pigs (both sexes and five age groups) from 38 representative intensive pig farms employing different parasite control strategies were examined for the presence of helminth ova and protozoan oocysts, cysts and/or trophozoites using standard techniques. Of the 3636 pigs sampled, 209 (5.7%) were infected with Trichuris suis, 189 (5.2%) with Ascaris, 91 (2.5%) with Oesophagostomum spp., 905 (24.9%) with coccidia (Eimeria spp. and/or Isospora suis) and 1716 (47.2%) with Balantidium coli. These infected pigs were mainly from farms without a strategic anti-parasite treatment regime. Concurrent infection of multiple parasites was common, and T. suis was the most common nematode infecting breeding, young and mature pigs. The results of the present investigation provide relevant 'base-line' data for assessing the effectiveness of control strategies against intestinal parasitism in intensively raised pigs in Guangdong Province, China.

Ethanol blocks both basic fibroblast growth factor- and carbachol-mediated neuroepithelial cell expansion with differential effects on carbachol-activated signaling pathways.

We have expanded neuroepithelial cells dissociated from the embryonic rat telencephalon in serum-free defined medium containing basic fibroblast growth factor (bFGF) in order to generate a model neuroepithelium to study the interaction of ethanol with both growth factor- and transmitter-stimulated proliferation. Ethanol blocked proliferation stimulated by bFGF and by carbachol, an agonist at muscarinic acetylcholine receptors, in a dose-dependent manner. In addition, ethanol attenuated autonomous expansion of neuroepithelial cells occurring following withdrawal of bFGF. The latter effect was associated with an increase in the number of apoptotic cells identified by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling labeling. We studied the effects of ethanol on carbachol-stimulated signaling pathways critical to its proliferative effects. Ethanol significantly reduced carbachol-stimulated Ca(2+) signaling, as well as Erk1/Erk2, Akt and cyclic AMP-response element-binding phosphorylations in a dose-dependent manner. Comparison of the potency of ethanol in attenuating carbachol-stimulated proliferation and signal transduction showed that mitogen-activated protein kinase phosphorylation was less sensitive to ethanol than the other parameters. The results indicate that ethanol's suppression of proliferation induced by carbachol in this model neuroepithelium likely involves multiple signaling pathways. These effects in vitro may help to explain the devastating effects of prenatal ethanol exposure in vivo, which contribute to the fetal alcohol syndrome.