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OBJECTIVE@#To investigate the effects of over-expression of miR-144 on invasion of SMMC-7721 cells and Toll-like receptor (TLR)/myeloid differentiation factor 88 (MyD88) pathway in hepatocellular carcinoma cells.@*METHODS@#The expressions of miR-144 was examined in normal human hepatocyte line HL-7702 and hepatocarcinoma cell line SMMC-7721 using realtime quantitative PCR (qRT-PCR). SMMC-7721 cells were divided into blank group, miR-144 NC group and miR-144 mimics group, and the expressions of miR-144 in each group were detected with qRT-PCR. Cell count kit-8 (CCK8) was used to assess the survival of SMMC-7721 cells, and the cell invasion was evaluated using Transwell assay. The expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and TLR/MyD88 pathway-related proteins in the cells were detected with Western blotting; the effect of 40 μ mol/L MyD88 inhibitor on TLR/MyD88 pathway-related proteins was examined in SMMC-7721 cells.@*RESULTS@#Compared with normal human hepatocytes, SMMC-7721 cells expressed a significantly lower level of miR-144 ( < 0.05). CCK-8 assay showed that test showed that miR-144 over-expression significantly decreased the cell survival rate ( < 0.05), lowered the number of invasive cells, and decreased the expression of MMP-2 and MMP-9 in SMMC-7721 cells ( < 0.05). The expressions of Toll-like receptor 4 (TLR4), MyD88, phosphorylated nuclear factor-kappa B (pNF-κB) and NF-κB protein decreased significantly in miR-144 mimics group and TJ-M2010-2 group ( < 0.05) and were comparable between the two groups ( > 0.05).@*CONCLUSIONS@#Overexpression of miR-144 decreases SMMC-7721 cell survival and invasion by inhibiting TLR/MyD88 pathway.
Subject(s)
Humans , Cell Line, Tumor , Liver Neoplasms , Matrix Metalloproteinase 2 , MicroRNAs , Myeloid Differentiation Factor 88 , NF-kappa B , Signal Transduction , Toll-Like ReceptorsABSTRACT
BACKGROUND:There are many preliminary studies on the survival, metaptosis, and correlation characteristics of human amniotic epithelial cel s after transplanted into the animals, but there are no reports on the quantitative analysis of the transplantation effect. OBJECTIVE:To make quantitative analysis on serum biochemical function of liver and the expression of human albumin in mice received passaged human amniotic epithelial cel s transplantation in spleen. METHODS:Forty nude mice were randomly divided into four groups (n=10 in each group):hepatectomy+cel transplantation 2 weeks group, hepatectomy+cel transplantation 4 weeks group, hepatectomy+normal saline group (treated with partial hepatectomy) and hepatectomy+cel transplantation group (transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen, and the blood were col ected at 2 and 4 weeks after transplantation). The mice in the hepatectomy+normal saline group were treated with splenic injection of 0.2 mL normal saline;the cel transplantation group did not receive hepatectomy, and transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen. The histological and morphological changes of the liver and spleen in each group as wel as the expressions of serum alanine aminotransferase, aspartate aminotransferase and human serum albumin in each group were detected, and the quantitative analysis of human serum albumin expression was performed. RESULTS AND CONCLUSION: There was no obvious morphological change after human amniotic epithelial cel s transplanted into the acute liver injury mice for 4 weeks, but specific cel s could be detected by histological method. The serum levels of alanine aminotransferase, aspartate aminotransferase and human serum albumin were improved obviously, and the human albumin could be detected in serum, the level of human albumin at 4 weeks after transplantation was significantly increased than 2 weeks after transplantation. Human amniotic epithelial cel s can survive for more than 4 weeks after transplanted into the liver injury mice, and can stil express partial characteristics and functions of hepatocyte-like cel s, improve the liver function, thus treating acute liver injury.
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OBJECTIVE@#To determine the therapeutic effect of laparoscopic splenectomy, perisoph-agogastric devascularization, and endoscopic variceal ligation (EVL) on patients with portal hypertension.@*METHODS@#We randomly divided 105 patients into 3 groups: 40 had endoscopic band ligation (the ligation group), 35 had splenectomy and perisoph-agogastric devascularization (the laparotomy group), and the other 30 had laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation (the combination group). Blood samples were analyzed preoperatively and postoperatively on day 1,3,and 7,including alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and directed bilirubin(DBIL). The length of stay, blood loss, operation time, anal exhaust time, azygos vein diameter, blood flow velocity and blood flow, recurrence of esophageal varices and rehaemorrhagia were compared.@*RESULTS@#Between the combination group and the laparotomy group, the serum levels of TbIL and Dbil had difference on 1st postoperative day(P<0.05). AST had difference on 7th postoperative day(P<0.05). The length of stay, blood loss, operation time, and anal exhaust time had significant difference(P<0.05). Among the combination group, the laparotomy group and the ligation group, the azygos vein blood flow before and after the treatment, recurrence of esophageal varices and rehaemorrhagia had no difference(P<0.05).@*CONCLUSION@#Laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation have less trauma, lower recurrence rate, fewer complications and rapid recovery, and may reduce the azygous vein blood flow. It can be used safely for portal hypertension.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Endoscopy , Methods , Esophageal and Gastric Varices , General Surgery , Hypertension, Portal , General Surgery , Laparoscopy , Methods , Ligation , Methods , Splenectomy , MethodsABSTRACT
Jaundice occurs in 19%-40% of the hepatocellular carcinoma (HCC) patients. HCC associated jaundice may be divided into hepatocellular and icteric types in terms of its underlying pathophysiology. The jaundice of icteric type is caused by obstruction of the bile duct through cancer embolus, blood clot, biliary sludge, tumor compression or infiltration. Jaundice and epigastric discomforts are the main clinical manifestations. In the present case, severe acute pancreatitis and acute cholangitis presenting as initial complaints of icteric type HCC were quite rare. A tumor located at the central lobe of the liver and a cancer embolus at the lower part of the common bile duct (CBD) were detected by CT scan. Curative resection of HCC with CBD exploration eradicated both the tumor and the embolus, and no recurrence was found after a 36 month follow-up.
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Objective To evaluate the in vitro differentiation of human amniotic epithelial cells (hAECs ) into hepatocyte-like cells. Methods Combined approach of dexamethasone, HGF, IGF and other cytokines were used to induce the differentiation of hAECs into hepatocyte-like cells. The induction lasted 2 weeks. During the induction, the expression of albumin ALB, CYP1A1, CYP1A2, IGFR, c-met and key functional genes related to liver cells as well as transcription factors HNF3, HNF4 and C/EBPa were monitored by RT-PCR. Time dependent changes of the surface marker colony ALB, AFP and CK18 were analyzed by cell flow cytometry. Results After the 2 week induction, the expressions of liver hepatocyte-like cell functional genes such as albumin, CYP1A1, CYP1A2, c-met, and transcription factors such as HNF3, HNF4, C/EBPa and HNF1 were observed. Six days after the induction, hAECs mainly were stained AFP+, and the positive rate was (15.1±2.1)%. While 10 days after the induction, part of the hAECs showed AFP+/ALB+ (6.5±1.4)%; and on 14th day, hAECs only showed ALB+, and the rate was (13.9±2.3)%. ALB+ cell increase indicated a gradual functional maturation from the hAECs to hepatocyte-like cells. Similaritly, the number of CK18+ cells in the whole population was also increased: On 10th day, the rate was (16.1±1.2)%; on 14th day, that was (21.3±4.6)%, which proved the above hypothesis of the trandifferentiation. By extending the induction time, the expression of functional genes increased gradually, and a maturing process of hAECs was detected by cell surface markers. Conclusion The differentiation of hAECs induced in vitro has the characteristics of hepatocyte-like cells.
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BACKGROUND:Human amniotic epithelial cells(AECs)are easy to obtain and can function as ideal seed cells for cell transplantation and tissue repair.Currently,marking and tracing of human AECs remains rarely reported.OBJECTIVE:To explore the efficiency of adeno-associated virus(AAV)vector-medicated green fluorescent protein(GFP)on in vitro cultured human AECs transfection.METHODS:Human amnion samples were harvested and trypsinized to isolate human AECs.The AECs were transfected with AAV-GFP,and the transfection efficiency was detected.RESULTS AND CONCLUSION:Human AECs were successfully primary cultured and passaged in vitro.AAV-GFP-transfected AECs stably and highly expressed GFP,with a transfection efficiency of 58%.