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Objective:To analyze the factors influencing prognosis of intrahepatic cholangiocarcinoma (ICC) after surgical resection.Methods:The clinical data of patients diagnosed with ICC and who underwent surgical resection from December 2015 to December 2019 at the Fifth Medical Center of PLA General Hospital were retrospectively analyzed. Of 39 patients who were included in this study, there were 23 males and 16 females, with age of (54.1±7.2) years old. The body mass index, hepatitis B virus infection status, tumor diameter, degree of differentiation, microvascular tumor thrombus, lymph node metastasis, and serum levels of carbohydrate antigen 19-9 (CA19-9) were analyzed as risk factors affecting postoperative recurrence and survival.Results:The median times to recurrence were significantly better in patients with a tumour length <5 cm (11 vs. 5 months), patients without microvascular tumor thrombus (54 vs. 6 months) and patients without lymph node metastasis (8 vs. 5 months) (all P<0.05). The median survival of patients with CA19-9≥100 U/ml was significantly shorter than that of patients with CA19-9<100 U/ml, (9 vs. 27 months, P<0.05). Tumor diameter>5 cm, microvascular tumor thrombus, lymph node metastasis, and CA19-9 ≥100 U/ml are risk factors affecting the recurrence time after ICC resection, CA19-9 ≥100 U/ml is a risk factor affecting survival time after ICC resection. Conclusion:Tumor diameter, microvascular tumor thrombus, lymph node metastasis and CA19-9 can be used to estimate the risk of ICC recurrence, and CA19-9 level can be used to estimate postoperative survival of ICC patients after resection.
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Objective To investigate the effects ofpravastatin on the expression ofmicroRNA-155 (miR-155) and the functions of lipopolysaccharide (LPS)-treated extravillous trophoblast cells.Methods In vitro cultured HTR-8/SVneo cells were divided into the following groups:control group,enhanced plasmid with green fluoscent protein (pEGFP)-miR-155 group (transfected with green fluorescent protein-tagged miR-155),LPS group (treated with 100 ng/mL of LPS),miR-155 inhibitor+LPS group,pravastatin+LPS group (treated with 100 ng/mL of LPS following pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin),and pravastatin+pEGFP-miR-155 group (transfected with pEGFP-miR-155 following pretreatment with 50 μ g/ml of pravastatin).Levels of miR-155 in HTR-8/SVneo cells treated with different strategies were measured by real-time polymerase chain reaction.Expression of phosphorylated JunB (p-JunB) and p-FosB proteins was analyzed by Western blotting.Migration,invasion and apoptosis of HTR-8/SVneo cells were also analyzed.All data were analyzed with t test.Results (1) Compared with the control group,HTR-8/SVneo cells in the pEGFP-miR-155 group were characterized by shorter migration distance [(274.70± 18.82) vs (181.00±8.62) μ m],less transmembrane cells [(123.00±4.36) vs (63.00±6.08)] and enhanced apoptosis [(5.40± 0.68)％ vs (9.27±0.68)％] (all P＜0.05).(2) Compared with the LPS group,the miR-155 inhibitor+LPS group showed longer migration distance of HTR-8/SVneo cells [(166.30±5.07) vs (242.00±18.07) μm,P＜0.05],more transmembrane cells [(71.67±6.12) vs (109.00±7.81),P＜0.05] and decreased cell apoptosis [(14.40±1.69)％ vs (6.23± 0.44)％,P＜0.05].(3) Expression of miR-155 at mRNA level in the LPS group was increased as compared with that of the control group (1.65 0.07 vs 0.79±0.12,P＜0.05).Compared with the LPS group,pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin decreased the expression of miR-155 at mRNA level [(1.14±0.10),(1.02±0.10),(0.74±0.15) and (1.140.02)],especially at the concentration of 50 μμ g/ml (all P＜0.05).(4) Expression ofp-JunB and p-FosB proteins in the control,LPS and pravastatin (50 μ g/ml)+LPS groups were (0.33 ±0.06) vs (0.37±0.07),(1.22±0.20) vs (0.80±-0.13),and (0.31 ±0.02) vs (0.21 ±0.05),respectively,showing higher expression level in both p-JunB and p-FosB proteins in the LPS group compared with that of the other two groups (all P＜0.05).(5) Compared with the LPS group,the pravastatin (50 μμ g/ml)+LPS group showed increased migration distance [(166.30±5.07) vs (246.80± 13.42) μ m,P＜0.05],increased numbers of transmembrane cells [(71.67 ± 6.12) vs (95.33 ± 2.73),P＜0.05] and decreased cell apoptosis [(14.40± 1.69) vs (6.05 ± 0.35)％,P＜0.05].(6) Compared with the pEGFP-miR-155 group,the pravastatin (50.00.00 μμ g/mL)+pEGFP-miR-155 group showed longer migration distance [(197.50± 13.86) vs (275.80± 13.63) μ m,P＜0.05],more transmembrane cells [(52.67±5.49) vs (125.00±6.66),P＜0.05] and lower rate of cell apoptosis [(8.90± 1.00) vs (5.05±0.35)％,P＜0.05].Conclusions Pretreatment of extravillous trophoblast cells with pravastatin can protect them from apoptosis and loss of migratory and invasive abilities through inhibiting the activation of AP-1 and down-regulating the expression of miR-155,which may be a mechanism that inhibits the development of preeclampsia.
ABSTRACT
ObjectiveTo investigate the relationship between levels of inflammatory cytokines, Ureaplasma infection in midtrimester amniotic fluid and spontaneous preterm delivery.MethodsFrom April 2009 to March 2012, a total of 1 865 pregnant women who underwent amniocentesis in midtrimester with known pregnant outcomes in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, among whom 43 cases who delivered spontaneously before 37 weeks, were classified as preterm group, and the control group consisted of 373 normal term delivery women selected from the other 1 785 cases who term delivered during the same period. Cytokines, including interleukin (IL)-10, IL-1β, IL-6, monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α) in amniotic fluid, were measured by MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel and analysis system for liquid phase chips.Ureaplasma DNA was detected by polymerase chain reaction. Sum-rank test was used to compare the difference of cytokines between the two groups and receiver-operating characteristic curve and Logistic analysis were used to analyze the value of cytokines to predict spontaneous preterm delivery.ResultsThe median maternal age in preterm group was higher than in the control group [34(24-49) vs 29(20-47) years] (Z=-3.107,P=0.002), but there was no significant difference in levels of the five cytokines among women with different age in control group(20-24, 25-29, 30-34 and≥35 years old) (allP>0.05). The levels of IL-1β and IL-6 were not significantly different between two groups (allP>0.05), but levels of IL-10, MCP-1 and TNF-α in the preterm group were much higher than in the control group [11.22(4.79-468.73) vs 7.87(0.93-26.62), 1 358.86(553.16-7 635.81) vs 1 137.98 (311.80-5 196.33), 9.78(5.72-106.51) vs 8.37(2.56-30.20) pg/ml, respectively;Z were-4.333,-2.820 and-3.390, allP<0.01]. The areas under receiver-operating characteristic curve of IL-10, MCP-1 and TNF-αwere 0.70, 0.63 and 0.66. The appropriate cut-off level of IL-10 in amniotic fluid for prediction of preterm birth was 9.06 ng/ml, with a sensitivity of 74.4% and a specificity of 57.6%, and the latter increased to 100.0% when combined with IL-10, MCP-1 and TNF-α, but the sensitivity declined to 16.3%.Ureaplasma was identified in only two preterm cases and none in the controls. However, the levels of the five cytokines in the two infected cases in preterm group were 2.12–43.00 times of the rest cases in the same group, and 2.26–60.00 times of the controls.ConclusionsAlthough IL-10, MCP-1 and TNF-α levels in midtrimester amniotic fluid are increased, it is not able to predict spontaneous preterm birth neither independently nor combined together. Ureaplasma infection rate in amnion cavity is low, but may related to parts of spontaneous preterm delivery.