1.Expression of the transmembrane emp24 domain-containing protein 4 in liver tissues of patients with hepatocellular carcinoma and its effects on biological behavior of hepatocellular carcinoma cells
Liyang WANG ; Wei HUANG ; Shuzhen WU ; Tao MA ; Zhaoxiu LIU ; Cuihua LU
Chinese Journal of Digestion 2022;42(10):667-674
Objective:To examine the expression of transmembrane emp24 domain-containing protein 4(TMED4) in liver tissue of patients with hepatocellular carcinoma, and to investigate the effects of TMED4 gene on the proliferation and migration of hepatocellular carcinoma cells and related molecular mechanisms. Methods:The expression of TMED4 at protein level in liver cancer tissue and paracancerous tissue of patients with hepatocellular carcinoma were detected by Western blotting and immunohistochemical stainning, and the correlation between its expression and clinicopathological features was analyzed. The effects of TMED4 overexpression or knockdown on proliferation, migration and healing ability of hepatocellular carcinoma cells in vitro and in vivo were determined by cell proliferation test, Transwell test, wound healing test and subcutaneous tumor formation in nude mice. The molecular mechanism of TMED4 in regulating the biological behavior of hepatocellular carcinoma cells was preliminarily explored by pathway analysis. Independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:The results of Western blotting showed that the expression of TMED4 at protein level in hepatocellular carcinoma tissue was lower than that in paracancerous tissue(0.52±0.29 vs. 0.83±0.22), and the difference was statistically significant( t=2.54, P=0.022). The results of immunohistochemical examination indicated that the expression of TMED4 at protein level in liver cancer tissue was lower than that in paracancerous tissue(5.46±3.37 vs. 7.58±3.08), and the difference was statistically significant( t=3.49, P<0.001). The expression of TMED4 at protein level was significantly correlated with vascular invasion and Barcelona clinic liver cancer stage( χ2=6.83 and 4.20, P=0.009 and 0.040). The results of cell proliferation assay showed that the absorbance value of SMMC-7721 cells in TMED4 overexpression group was lower than that in control group(1.38±0.05 vs. 2.37±0.08), while the optical density value of HepG2 in TMED4 knockdown group was higher than that in control group(0.76±0.04 vs. 0.54±0.01), and the differences were statistically significant( t=18.23 and 8.85, both P<0.001). The results of Transwell test showed that the number of migrated SMMC-7721 cells in TMED4 overexpression group was less than that in control group(286.30±13.01 vs. 439.70±12.34), while the number of migrated HepG2 cells in TMED4 knockdown group was higher than that in control group(249.00±6.00 vs. 160.00±6.56), and the differences were statistically significant( t=14.81 and 17.34, both P<0.001). The wound healing test showed that the healing rate of SMMC-7721 cells in TMED4 overexpression group was lower than that in control group((0.21±0.01)% vs.(0.45±0.01)%), the healing rate of HepG2 cells in TMED4 knockdown group was higher than that in control group((0.46±0.01)% vs.(0.20±0.01)%), and the differences were statistically significant( t=200.10 and 30.46, both P<0.001). The results of subcutaneous tumor formation assay in nude mice showed that the growth rate of cells in TMED4 overexpression group was slower than that in control group. After cell inoculation for 6 weeks, the subcutaneous tumor volume of mice in TMED4 overexpression group was smaller than that in control group(27.36 mm 3(138.70 mm 3) vs. 1 741.62 mm 3(1 783.39 mm 3)), the tumor weight was lower than that in control group(0.06 g(0.14 g) vs. 1.46 g(1.09 g)), and the differences were statistically significant(both Z=-2.31, both P<0.001). The results of Western blotting showed that the expression of Snail at protein level in SMMC-7721 cells of the TMED4 overexpression group was lower than that of the control group(0.32±0.01 vs. 0.90±0.03), the protein level of Snail in HepG2 cells of TMED4 knockdown group was higher than that of control group(1.03±0.01 vs. 0.97±0.01), and the differences were statistically significant( t=28.49 and 12.31, both P<0.001). The results of real time fluorescent quantitative polymerase chain reaction showed that the expression of Snail at mRNA level in SMMC-7721 cells of TMED4 overexpression group was lower than that of control group(0.13±0.05 vs. 1.00±0.15), the expression of Snail at mRNA level in HepG2 cells of TMED4 knockdown group was higher than that of control group(1.25±0.32 vs. 0.21±0.14), and the differences were statistically significant( t=9.62 and 5.10, P<0.001 and P=0.007). Conclusion:TMED4 may affect the proliferation and migration of hepatocarcinoma cells by regulating the expression of Snail, and which is expected to become a potentially therapeutic target for hepatocellular carcinoma.
2.The effect of hyperlipidemia on perioperative outcomes of rectal cancer patients
Xin WU ; Rongkang KUANG ; Liyang LIU ; Haitao YANG ; Jing TAO
Chinese Journal of Geriatrics 2021;40(10):1295-1298
Objective:To examine the influence of hyperlipidemia on perioperative outcomes of rectal cancer patients.Methods:From June 2016 to June 2019, 86 patients who had received laparotomy or laparoscopic radical resection of rectal cancer at the Department of General Surgery of Nanjing Jiangbei People's Hospital were selected as research subjects.Preoperative blood lipids were measured.Based on lipid levels, patients were divided into the hyperlipidemia group(46 cases)and the normal blood lipid group(40 cases). Operative time, intraoperative blood loss, postoperative recovery outcomes, incidence of anastomotic fistula, and incidences of incision fat liquefaction and incision infections were compared between the two groups.SPSS21.0 was used for data analysis.Results:Compared with the normal lipid group, the operative time of the hyperlipidemia group was significantly longer and the amount of intraoperative blood loss was higher in the hyperlipidemia group, and the differences were statistically significant.Compared with the normal lipid group, the hyperlipidemia group showed a longer time to first postoperative flatus and to first postoperative oral intake, delayed drainage tube removal and ambulation, and longer hospital stays, with statistically significant differences.There were 17 cases of postoperative complications in the hyperlipidemia group(37.0%)and 4 cases in the normal lipid group(10.0%), and the difference was statistically significant( χ2=7.027, P=0.008). Conclusions:Hyperlipidemia has adverse effects on surgical outcomes and postoperative recovery for rectal cancer patients.
3.Secalonic acid D induces cell apoptosis in both sensitive and ABCG2-overexpressing multidrug resistant cancer cells through upregulating c-Jun expression.
Hong ZHANG ; Liyan HUANG ; Liyang TAO ; Jianye ZHANG ; Fang WANG ; Xu ZHANG ; Liwu FU
Acta Pharmaceutica Sinica B 2019;9(3):516-525
Secalonic acid D (SAD) could inhibit cell growth in not only sensitive cells but also multidrug resistant (MDR) cells. However, the molecular mechanisms need to be elucidated. Here, we identified that SAD possessed potent cytotoxicity in 3 pairs of MDR and their parental sensitive cells including S1-MI-80 and S1, H460/MX20 and H460, MCF-7/ADR and MCF-7 cells. Furthermore, SAD induced cell G2/M phase arrest the downregulation of cyclin B1 and the increase of CDC2 phosphorylation. Importantly, JNK pathway upregulated the expression of c-Jun in protein level and increased c-Jun phosphorylation induced by SAD, which was linked to cell apoptosis c-Jun/Src/STAT3 pathway. To investigate the mechanisms of upregulation of c-Jun protein by SAD, the mRNA expression level and degradation of c-Jun were examined. We found that SAD did not alter the mRNA level of c-Jun but inhibited its proteasome-dependent degradation. Taken together, these results implicate that SAD induces cancer cell death through c-Jun/Src/STAT3 signaling axis by inhibiting the proteasome-dependent degradation of c-Jun in both sensitive cells and ATP-binding cassette transporter sub-family G member 2 (ABCG2)-mediated MDR cells.
5.Brazilein induced cells apoptosis in human lung cancer A549 cells and its effects on endoplasmic reticulum stress
Liyang TAO ; Jianying LI ; Jianye ZHANG
Chongqing Medicine 2015;(30):4180-4182
Objective To study the apoptotic effect of brazilein on human lung cancer A549 cells and endoplasmic reticulum stress .Methods The cytotoxic activity was tested by MTT assay in A549 cells .The flow cytometry was used to detect apoptosis . Western blotting was performed to detect GRP78 and cyto c protein expression .Results The IC50 values of brazilein against A549 cells was (5 .36 ± 0 .62)μmol/L .After treatment with 0 ,5 ,10 and 20 μmol/L brazilein for 48 h ,the percent of apoptosis was (1 .15 ± 0 .32)% ,(19 .61 ± 4 .52)% ,(30 .18 ± 6 .35)% and (39 .48 ± 7 .44)% respectively .There was significant difference among the different treatment (P< 0 .05) .Compared with control group ,the protein expression of GRP78 and cytosolic cyto c was in‐creased after 5 ,10 and 20 μmol/L brazilein treated for 48 h .Conclusion Brazilein induced apoptosis in human lung cancer A549 cells though endoplasmic reticulum stress pathway .
6.Clear cell chondrosarcoma of cricoid cartilage: report of a case.
Yuanyuan YAO ; Miaoxia HE ; Shicai CHEN ; Ting FENG ; Liyang TAO ; Dalie MA ; Jianming ZHENG ; Minghua ZHU
Chinese Journal of Pathology 2014;43(1):42-43
Chondroblastoma
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pathology
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Chondroma
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pathology
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Chondrosarcoma
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metabolism
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pathology
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surgery
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Cricoid Cartilage
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Diagnosis, Differential
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Humans
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Laryngeal Neoplasms
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metabolism
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pathology
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surgery
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Laryngectomy
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Lymph Node Excision
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Male
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Middle Aged
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Osteoblastoma
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pathology
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Osteosarcoma
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pathology
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S100 Proteins
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metabolism
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Sarcoma, Clear Cell
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metabolism
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pathology
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surgery
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Vimentin
;
metabolism
7.Interventional revascularization of the lower extremity arteries with complex arteriosclerosis obliterans
Jingyu LI ; Tao LIU ; Junliang LU ; Liyang XU
Chinese Journal of Radiology 2011;45(10):960-963
ObjectiveTo explore the methods and effectiveness of interventional revascularization of complex arteriosclerosis obliterans (ASO)of lower extremity arteries according to their imaging characteristics.Methods Seventy-eight patients with lower extremity ASO complex lesions classified as TASC Ⅱ C/D ( n =68 ) and TASC Ⅱ B ( n =10) underwent antigrade or combined antigrade-retrograde subintimal angioplasty.Their clinical situations were Fontaine stage Ⅲ/Ⅳ or severe stage Ⅱ.All the long occlusion,flush occlusion,multiple occlusion,popliteal artery occlusion,below knee artery occlusion and aortoiliac artery occlusion were identified as complex lesion imaging features and as the indication of interventional treatment if only there were visible outflow vessels and suitable puncture site.Statistical analysis was used to compare ankle-brachial index (ABI) pretreatment and post-treatment by t test.Results Successful revascularization was achieved in 73 patients technically.No obvious complications occurred.Ischemia symptoms improved quickly after accomplishment of recanalization.Average ABI increased from 0.45 ±0.07 to 0.76 ±0.11 after the treatment(t =- 19.78,P <0.01 ).Symptoms in 5 patients who failed to get arterial recanalization were stable.Follow up of 6 to 12 months in 47 patients showed stable improvement in 22 of them.Conclusion It is more practical to choose and expand application of interventional treatment for complex ASO according to imaging characteristics of lesions than according to TASC classification.
8.Percutaneous transsplenic varices embolization for upper gastrointestinal bleeding
Qiang ZHANG ; Jingyu LI ; Junliang LU ; Liyang XU ; Tao LIU
Chinese Journal of Radiology 2010;44(11):1194-1196
Objective To investigate the value of percutaneous transsplenic varices embolization (PTSVE) for treatment of upper gastrointestinal bleeding. Methods Twenty cases with liver cirrhosis and portal hypertension suffered upper gastrointestinal bleeding. PTSVE was administered to them with hardener and coils. Among them, 8 cases had massive hepatocellular carcinoma (HCC) in right lobe; 10 cases with hepatocellular carcinoma had portal vein tumor thrombus and occlusion; the other two cases with liver cirrhosis had portal vein thrombosis. All of these cases were not suitable for percutaneous transhepatic varices embolization (PTHVE) . PTSVE was performed under the guidance of fluoroscopy. Results Technical success was achieved in 18 patients. A total of 35 gastric coronary veins were embolized. In all these cases, upper gastrointestinal bleeding stopped after PTSVE. There was no recurrence within 1 month follow-up. No serious complication occurred. Conclusion PTSVE is a safe and efficient alternative treatment for upper gastrointestinal bleeding, especially for cases with portal vein occlusion or with massive HCC in right lobe of liver.
9.Application of bidirectional subintimal angioplasty in atherosclerotic occlusion of lower extremities
Junliang LU ; Jingyu LI ; Qiang ZHANG ; Liyang XU ; Tao LIU
Journal of Interventional Radiology 2006;0(10):-
Objective To discuss the therapeutic efficacy of bidirectional subintimal angioplasty in the treatment of atherosclerotic occlusive of lower limbs.Methods Five patients with long segment of obstructed artery in lower limb were enrolled in the study.Of five patients,occlusion of the lower segment of abdominal aorta and bilateral iliac artery was seen in one,occlusion of iliac artery in 2 and occlusion of superficial femoral artery in the remaining two.Antegrade subintimal angioplasty procedure was unsuccessful in all five patients as the wire could not be placed into the true lumen,so retrograde subintimal angioplasty by puncturing the distal segment of the occlusive artery was employed.Through the newly created channel the retrograde guide wire was manipulated to be pulled out of vessel through the antegrade catheter.The subintimal tract was dilated with angioplasty balloon and the stent implantation was performed.Results Bidirectional subintimal angioplasty was successfully accomplished in all five patients,with a total of nine stents being implanted.Conclusion Bidirectional subintimal angioplasty is a safe and effective procedure for bringing the subintimal recanalization to success,this technique can be regarded as a remedial measure when unidirectional subintimal angioplasty ends up in failure.

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