To explore a simple and easy-to-learn procedure for the isolation of human quiescent hepatic stellate cells (HSCs) that requires no advanced training. Thus reducing costs and increasing efficiency. This protocol will provide sufficient primary cells with minimal contaminants for future basic research on diseases associated with human HSCs. Normal liver tissues were isolated from patients undergoing hepatic hemangioma resection, and a single cell suspension of these tissues was prepared using the Gentle MACS tissue processor. By using this method, the difficulty of the procedure was reduced, fewer cells were lost during the preparation treatments, and the maximal activity of single cells was maintained. Following preparation of the cell suspension, the HSCs were further isolated using a Nycodenz density gradient. Cell viability was examined by trypan blue staining, and the purity of the quiescent human HSCs was determined by autofluorescence and oil red O staining. Activated and quiescent human HSCs were identified using immunofluorescence and Western blotting. The cell cycle distribution in activated and quiescent human HSCs was analyzed by flow cytometry.The recovery rate of the HSCs was approximately (2.1 ± 0.23) × 10
6
of tissue, with 94.43 ± 1.89% cell viability and 93.8 ± 1.52% purity. The technique used in this study is a simple, high-yield, and repeatable method for HSC isolation that is worthy of recommendation.
Objective To describe the surgical procedures of laparoscopic caudate lobectomy and analyze its clinical efficiency for treating cancer. Methods Twelve consecutive patients of hepatocellular carcinoma, hepatic hemangioma, and focal nodular hyperplasia who received laparoscopic caudate lobectomy in Qilu Hospital of Shandong University from January 2013 to January 2017 were included in this study. The clinical data, intraoperative parameters, and postoperative outcomes were assessed. Results All 12 patients received totally laparoscopic technique. The operative time was 140.8 ± 95.34 minutes. The average estimated blood loss was 97.92 ± 90.54 ml, and no blood transfusions were required. The mean duration of hospital stay was 9.17 ± 2.88 days. There was no perioperative complication or patient mortality in this series. Conclusions Laparoscopic caudate lobectomy is safe and feasible in the selected patients.
We aim to investigate the effects of postoperative adjuvant transarterial chemoembolization (TACE) on survival and recurrence in hepatocellular carcinoma (HCC) patients after radical resection. A total of 320 HCC patients underwent radical resection between January 2010 and January 2014 in Qilu Hospital, Shandong University were divided into 4 groups according to the frequency of postoperative adjuvant TACE. Patients were further stratified into subgroups (tumor diameter ≤5 or >5 cm) with low or high risk factors for recurrence or death. A low risk factor for recurrence or death was defined as Edmondson grade I/II without microvascular invasion (MiVI), while a high risk factor was defined as Edmondson grade III/IV or with MiVI. Survival data and recurrence rates were compared using the Kaplan–Meier method. Uni- and multivariate analyses were based on the Cox proportional analysis. Compared to those received no TACE, patients underwent 2 (log-rank, χ2 = 9.054, P = .003) or 3 (log-rank, χ2 = 4.228, P = .04) TACE showed delayed recurrence. Patients received 2 or 3 TACE showed extended overall survival (OS) compared with the other patients. No statistical differences were found between all the disease-free survival (DFS) and OS in low-risk subgroups. In the patients of the high-risk subgroup with a tumor diameter of ≤5, those received 2 TACE showed delayed recurrence compared with those received no TACE, and TACE (twice or thrice) can improve OS. For those of the high-risk subgroup with a tumor diameter of >5, TACE (twice or thrice) can delay recurrence and improve OS. Adjuvant TACE (twice or thrice) after radical resection is beneficial for HCC patients with poor differentiation and MiVI, especially for those with a tumor diameter of >5 cm.
PurposeTo determined KIAA1199 expression and investigate its correlation with the clinicopathologic data and prognosis of hepatocellular carcinoma (HCC), as well as markers of epithelial-mesenchymal transition (EMT); N-cadherin, E-cadherin and vimentin.Materials and methodsWestern blot, quantitative real-time PCR, and immunohistochemical staining were used to measure KIAA1199 expression in human HCC specimens. Subsequently, the correlation between KIAA1199 expression and the pathological characteristics of HCC patients was analyzed. Univariate and multivariate analyses were used to explore the risk factors associated with disease-free survival (DFS) and overall survival (OS).ResultsKIAA1199 expression was remarkably increased in hepatocellular carcinoma tissues compared to paracarcinomatous tissues. This phenomenon was accompanied by aberrant expression of EMT-associated markers. In addition, high KIAA1199 expression was associated with severe pathological symptoms, low DFS, and low OS. Results of the multivariate analysis showed that KIAA1199 expression may be an independent predictor of low disease-free survival and OS of HCC patients.ConclusionKIAA1199 overexpression in HCC patients is associated with aberrant expression of EMT-associated markers and severe clinicopathological symptoms, and thus may function as a marker of poor prognosis in HCC.
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