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Objective:To investigate the relationship between the expression of inhibin subunit Beta A (INHBA) and the clinicopathological data and its effect on proliferation, invasion and metastasis of gastric cancer cells and its possible mechanisms.Methods:Using Gene Expression Profiling Interactive Analysis (GEPIA) public database to verify the expression of INHBA mRNA in gastric cancer and adjacent tissues and its relationship with pathological stage and prognosis; the relationship between INHBA and clinical prognosis was analyzed using the Kaplan-Meier Plotter online database. Immunohistochemistry was used to confirm the correlation between protein expression level of INHBA in gastric cancer and adjacent tissues and clinical pathological staging. In vitro, the cell proliferation was detected by tetrazolium salt (MTT) method; the cell migration ability was detected by scratch test, and cell invasion and metastasis ability was detected by transwell chamber assay. The expression of INHBA and epithelial-mesenchymal transition (EMT) related proteins was detected by Western blot. Results:The GEPIA database and Kaplan-Meier Plotter online database analysis showed that INHBA mRNA was highly expressed in various cancer tissues and significantly higher in gastric cancer tissues than normal tissues ( P<0.05). The high expression of INHBA mRNA was associated with clinical stage and poor prognosis of gastric cancer ( P<0.05). The results of immunohistochemistry showed that the staining score of INHBA in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.05), and its expression level was correlated with clinical tumor node metastasis (TNM) stage ( P<0.05). The results of MTT, scratch test and transwell chamber showed that INHBA overexpression could promote the proliferation, migration, invasion and metastasis of gastric cancer cells, while interference with INHBA expression could inhibit the proliferation, migration, invasion and metastasis of gastric cancer cells; Western blot results showed that the expression of CDH1 was down regulated and the expression of CDH2 was up-regulated after INHBA overexpression. The expression of CDH1 was up-regulated and the expression of CDH2 was down-regulated after INHBA overexpression was inhibited. Conclusions:INHBA is highly expressed in gastric cancer tissues compared with adjacent tissues. The expression level of INHBA is related to tumor progression and poor prognosis. INHBA can promote the proliferation, migration and invasion of gastric cancer MGC-803 cell line and its mechanism may be related to INHBA promoting cell EMT.
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Objective: To investigate the effect of DSA-guided transnasal ileus tube implantation for treatment of refractory adhesive ileus. Methods Totally 50 patients with refractory adhesive ileus were treated with transnasal ileus tube implantation under the guidance of DSA. The time of intubation, time of relieving abdominal pain and distention, the flow of negative pressure, the time of anal exhaust and defecation were counted. The length of tube entry was counted on the 3rd and 6th day after operation, and the position of tube head was observed with abdominal X-ray films. Results All 50 patients were successfully catheterized in one time, the average catheterization time was (22.57±6.93)min.The patients were well tolerated, no gastrointestinal bleeding nor perforation occurred. Forty-five patients (45/50, 90%) were relieved of abdominal pain and distention, and the average time was (9.64±8.33)h. The negative pressure drainage volume was (1 500±450)ml on the first day, (750±120)ml on the second day and (257±112)ml on the third day. On the third day after operation, the average length of the tube was (195.97±14.63)cm, and abdominal X-ray films showed that the head of the tube located in the distal jejunum (Group 3). On the sixth day after operation, the average length of the tube was (240.55±17.65)cm, and the head of the tube located in the distal ileum. The average time of anal exhaust was (2.80±1.01)d, and of defecation was (3.52±1.26)d. Conclusion: DSA-guided transnasal ileus tube implantation is effective for treatment of refractory adhesive ileus, which can significantly relieve clinical symptoms and restore the physiological function of intestinal tract.
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ObjectiveTo investigate the expression features and clinical significance of the microRNA-888 (miRNA-888) gene family members in hepatocellular carcinoma (HCC). MethodsA total of 72 patients with pathologically confirmed HCC who were treated in The First Affiliated Hospital of Hunan University of Chinese Medicine from January 2012 to December 2017 were enrolled, and 72 liver tissue samples collected from healthy volunteers were enrolled as normal control group. Quantitative real-time PCR and in situ hybridization were used to measure the expression of miRNA-888 family members (miRNA-888, miRNA-891a, miRNA-891b, miRNA-892a, and miRNA-892b) in HCC tissue, and the association of the expression of miRNA-888, miRNA-891b, and miRNA-892a with the clinicopathological features of HCC was analyzed. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsHCC tissue had significantly higher expression of miRNA-888, miRNA-891b, and miRNA-892a than normal liver tissue (miRNA-888: 2.53±0.75 vs 0.46±0.08, t=14.02, P＜0.001; miRNA-891b: 2.26±0.38 vs 1.19±0.21, t=7.75, P＜0.001; miRNA-892a: 5.44±1.01 vs 1.06±0.30, t=35.27, P＜0001). According to the results of in situ hybridization, the miRNAs of miRNA-888, miRNA-891b, and miRNA-892a were mainly located in the cell nucleus of HCC tissue, and there were significant differences in the expression of these miRNAs between HCC tissue and normal tissue (miRNA-888: 3.91±0.92 vs 1.21±0.42, t=22.65, P＜0.001; miRNA-891b: 2.92±0.76 vs 0.83±0.21, t=2292, P＜0001; miRNA-892a: 3.81±0.99 vs 1.30±0.32, t=20.47, P＜0001). There were significant differences in the expression of miRNA-888 and miRNA-891b between the patients with different histological grades and clinical stages (χ2=6.25, 4.44, 4.76, and 6.05, all P＜0.05), and there was a significantly higher expression of miRNA-892a in stage III/IV tumors (χ2=8.50, P＜0.001). ConclusionThere are significant increases in the expression of miRNA-888, miRNA-891b, and miRNA-892a in HCC tissue, and such increases are closely associated with the malignancy of HCC.
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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.
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Adult , Female , Humans , Male , Middle Aged , Endoscopy , Methods , Esophageal and Gastric Varices , General Surgery , Hypertension, Portal , General Surgery , Laparoscopy , Methods , Ligation , Methods , Splenectomy , MethodsABSTRACT
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%.