OBJECTIVETo investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma (HCC).

METHODSExpression levels of CCL20 mRNA in the normal human liver cell line L-02, and human hepatocellular carcinoma cell lines Hep3B, Huh7 and HepG2 were quantified by using SYBR green real time PCR. CCL20 secretions from these cell lines were quantified by using ELISA. The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay. Pre-therapy serum levels of IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-17, IL-23, IFN-γ, TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA. All the patients were chronic hepatitis B virus associated HCC, and 51 cases were those with vascular invasion and metastasis (metastasis group) and 42 cases were not (non-metastasis group). CCL20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients, among them, 20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis. The CCL20 expression was further determined by immunohistochemistry.

RESULTSThe HCC cell lines expressed and secreted higher amount of CCL20, which effectively recruited CCR6(+) T cells. Pre-therapy serum levels of CCL20 in 93 HCC patients were (38.2 ± 28.4)pg/ml, significantly increased than those with benign hepatic hemangiomas [(7.8 ± 17.8)pg/ml, P < 0.01]. In addition, the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients (r = 0.32, P = 0.0018). CCL20 was dominantly expressed in the cytoplasm in HCC cells, and it was also expressed by some infiltrating immune cells. The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues (P < 0.05). Multivariate logistic regression analysis showed that serum levels of IL-17 and CCL20 were independent risk factors of metastasis in HCC patients (P < 0.05 for both). CCL20 mRNA showed no statistically significant differences between patients with metastasis and without metastasis in both tumor tissues and tumor-adjacent tissues (P > 0.05 for both). But the patients with metastasis showed significantly higher expressions of CCR6 both in their tumor [5.75 (1.79, 19.13)]and tumor-adjacent tissues [7.99 (4.49, 19.54)] than those with non-metastasis [1.69 (0.76, 2.87) and 3.58 (1.84, 4.32), P < 0.05 for both].

CONCLUSIONCCL20/CCR6/Th17 axis may promote vascular invasion and metastasis hepatocellular carcinoma.

Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; metabolism ; Chemokine CCL20 ; metabolism ; Humans ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-23 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocytes, Mononuclear ; Liver Neoplasms ; metabolism ; RNA, Messenger ; Th17 Cells ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the impact of overweight on the prognosis of patients with acute mild ischemic stroke or moderate-high risk transient ischemic attack(TIA).Methods A total of 366 patients with acute mild ischemic stroke or moderate-high risk TIA who carried CYP2C19 loss-of-function alleles were included in the study.Baseline data were collected at admission,and clinical outcomes were collected within 3 months post-onset(primary outcomes:stroke recurrences within 3 months post-onset;secondary outcomes:composite outcome of stroke recurrence or death within 3 months post-onset,vascular events,and quality of life within 3 months).Kaplan-Meier method was used to plot the cumulative incidence curve of outcomes.Multivariate Cox proportional hazards mod-el and multivariate Logistic regression model were used to evaluate the relationship between over-weight and clinical outcomes within 3 months post-onset in patientswith ischemic stroke or TIA.The integrated discrimination improvement index(IDI)and net reclassification improvement index(NRI)were calculated to assess the predictive value of adding body mass index(BMI,whether o-verweight or not)based on traditional models for predicting the prognosis of acute ischemic stroke or TIA.Results After 3 months of follow-up,28 patients had stroke recurrence,1 patient died,and 31 patients had vascular events.The Kaplan-Meier cumulative incidence curve showed that the cu-mulative incidences of stroke recurrence,stroke recurrence or death,and vascular events after 3 months of onset were lower in overweight patients compared with non-overweight patients(Log-rank P＜0.05).Multivariate analysis showed that compared with non-overweight patients,overweight pa-tients had significantly reduced risks of stroke recurrence within 3 months(HR=0.24;95％CI,0.08 to 0.71),composite outcome of stroke recurrence or death(HR=0.24;95％CI,0.08 to 0.69),and vascular events(HR=0.22;95％CI,0.07 to 0.63),and significantly improved qual-ity of life within 3 months(OR=0.39;95％CI,0.20 to 0.76).The IDI and NRI calculations showed that compared with traditional models,the new model adding BMI(whether overweight)had significantly improved predictive ability for the prognosis of acute ischemic stroke or TIA.Conclu-sion Overweight may be a protective factor for the prognosis of TIA patients with acute mild ische-mic stroke or moderate-high risk who carry CYP2C19 loss-of-function alleles.Compared with non-o-verweight patients,overweight patients have reduced risks of stroke recurrence,composite outcome of stroke recurrence or death,and vascular events within 3 months post-onset,and improved quality of life within 3 months post-onset.

Objective To investigate the impact of overweight on the prognosis of patients with acute mild ischemic stroke or moderate-high risk transient ischemic attack(TIA).Methods A total of 366 patients with acute mild ischemic stroke or moderate-high risk TIA who carried CYP2C19 loss-of-function alleles were included in the study.Baseline data were collected at admission,and clinical outcomes were collected within 3 months post-onset(primary outcomes:stroke recurrences within 3 months post-onset;secondary outcomes:composite outcome of stroke recurrence or death within 3 months post-onset,vascular events,and quality of life within 3 months).Kaplan-Meier method was used to plot the cumulative incidence curve of outcomes.Multivariate Cox proportional hazards mod-el and multivariate Logistic regression model were used to evaluate the relationship between over-weight and clinical outcomes within 3 months post-onset in patientswith ischemic stroke or TIA.The integrated discrimination improvement index(IDI)and net reclassification improvement index(NRI)were calculated to assess the predictive value of adding body mass index(BMI,whether o-verweight or not)based on traditional models for predicting the prognosis of acute ischemic stroke or TIA.Results After 3 months of follow-up,28 patients had stroke recurrence,1 patient died,and 31 patients had vascular events.The Kaplan-Meier cumulative incidence curve showed that the cu-mulative incidences of stroke recurrence,stroke recurrence or death,and vascular events after 3 months of onset were lower in overweight patients compared with non-overweight patients(Log-rank P＜0.05).Multivariate analysis showed that compared with non-overweight patients,overweight pa-tients had significantly reduced risks of stroke recurrence within 3 months(HR=0.24;95％CI,0.08 to 0.71),composite outcome of stroke recurrence or death(HR=0.24;95％CI,0.08 to 0.69),and vascular events(HR=0.22;95％CI,0.07 to 0.63),and significantly improved qual-ity of life within 3 months(OR=0.39;95％CI,0.20 to 0.76).The IDI and NRI calculations showed that compared with traditional models,the new model adding BMI(whether overweight)had significantly improved predictive ability for the prognosis of acute ischemic stroke or TIA.Conclu-sion Overweight may be a protective factor for the prognosis of TIA patients with acute mild ische-mic stroke or moderate-high risk who carry CYP2C19 loss-of-function alleles.Compared with non-o-verweight patients,overweight patients have reduced risks of stroke recurrence,composite outcome of stroke recurrence or death,and vascular events within 3 months post-onset,and improved quality of life within 3 months post-onset.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.