Blockade of the deubiquitinating enzyme USP48 degrades oncogenic HMGA2 and inhibits colorectal cancer invasion and metastasis.

HMGA2, a pivotal transcription factor, functions as a versatile regulator implicated in the progression of diverse aggressive malignancies. In this study, mass spectrometry was employed to identify ubiquitin-specific proteases that potentially interact with HMGA2, and USP48 was identified as a deubiquitinating enzyme of HMGA2. The enforced expression of USP48 significantly increased HMGA2 protein levels by inhibiting its degradation, while the deprivation of USP48 promoted HMGA2 degradation, thereby suppressing tumor invasion and metastasis. We discovered that USP48 undergoes SUMOylation at lysine 258, which enhances its binding affinity to HMGA2. Through subsequent phenotypic screening of small molecules, we identified DUB-IN-2 as a remarkably potent pharmacological inhibitor of USP48. Interestingly, the small-molecule inhibitor targeting USP48 induces destabilization of HMGA2. Clinically, upregulation of USP48 or HMGA2 in cancerous tissues is indicative of poor prognosis for patients with colorectal cancer (CRC). Collectively, our study not only elucidates the regulatory mechanism of DUBs involved in HMGA2 stability and validates USP48 as a potential therapeutic target for CRC, but also identifies DUB-IN-2 as a potent inhibitor of USP48 and a promising candidate for CRC treatment.

Editor's Choice - Clinical Efficacy of Venastent - A Novel Iliac Vein Stent for Non-Thrombotic Iliac Vein Lesions: A Multi-Centre Randomised Controlled Trial.

OBJECTIVE: To determine the efficacy of Venastent - a novel iliac vein stent for non-thrombotic iliac vein lesions (NIVLs). METHODS: From October 2018 to January 2021, 256 NIVL patients were recruited at 19 Chinese hospitals. A randomised controlled trial was conducted to compare the efficacy of the new iliac vein stent-Venastent (Tianhong China) with Zilver stent (Cook USA). All patients were allocated randomly to two groups: the experimental group patients used Venastent, while the control group received the Zilver stent. The trial was registered in Chinese Clinical Trial Registry (ChiCTR2200057851). RESULTS: A total of 123 patients in the experimental group and 122 patients in the control group had a full set of data collected (p = ns). The technical success rate was 100% (n = 245/245). The patency rate was 100% (n = 123/123) in the experimental group and 98.4% (n = 120/122) in control group one year after operation (p = ns). The lower extremity swelling remission rate was 79.1% (n = 87/110) in the experimental group and 78.4% (n = 91/116) in the control group (p = ns). The lower extremity pain relief rate was 68.8% (n = 50/80) in the experimental group and 77.2% (n = 71/92) in the control group (p = ns). The ulcer healing rate was 90% (n = 18/20) in the experimental group and 87% (n = 20/23) in the control group (p = ns). There was no difference in stent re-stenosis or clinical remission between the two groups. CONCLUSION: The new iliac vein stent, Venastent, had a comparable high patency rate and safety profile as the Zilver stent (Cook) in NIVLs patients. Venastent significantly reduced symptoms of chronic venous disease.

Clinical Results and Aortic Remodeling After Endovascular Treatment for Complicated Type B Aortic Dissection With the "Fabulous" Stent System.

OBJECTIVE: To report the clinical outcomes and aortic remodeling after the implantation of a self-developed, biomechanically optimized, two-stage thoracic stent system named Fabulous. BACKGROUND: Given the efficacy of the PETTICOAT concept, the benefits of Fabulous and the behavior of remodeling in different segments need further investigation. METHODS: This is a prospective and multicenter study. From 2017 to 2019, 145 patients (mean age, 56.6 years; 88.3% male) from 14 centers were included in this cohort. The clinical results and core laboratory results were from a central electronic data capture system. Computed tomographic angiography was performed preoperatively, 1 month, 6 months and yearly thereafter and was used for volumetric analysis by 3mensio (Bilthoven, The Netherlands). After the 1-year follow-up, 97.2 and 87.6% of the clinical and imaging results of the eligible patients were available. RESULTS: Both stent grafts and bare stents were successfully delivered in place in 100% of the patients. The 30-day mortality and 1-year freedom from all-cause mortality were 2.1 and 96.6%, respectively. The incidence of entry flow was 11.7% at 30 days and 6.2% at 365 days. No cases of stent-induced new entry (SINE) or reintervention were observed. After the 1-year follow-up, the true lumen/overall volume ratio reached 88%. The following subdivided segment volume changes were recorded: stent graft segment TL +56%; FL -92%, bare stent segment TL +32%; FL -75%, and there were no significant changes in the visceral segment. CONCLUSIONS: These outcomes indicated that there were favorable clinical benefits of Fabulous stent system. This device achieved a low short-term mortality and a low incidence of reintervention. In addition, patients undergoing Fabulous stent system implantation showed remodeling both on descending aorta and on the distal aorta. The volume changes in the TL and FL varied in the different segments. The long-term follow-up is still ongoing.

G protein-coupled receptor 39 activation alleviates oxidized low-density lipoprotein-induced macrophage inflammatory response, lipid accumulation and apoptosis by inducing A20 expression.

G protein-coupled receptor 39 (GPR39) agonist weakens oxidized low-density lipoprotein (ox-LDL)-induced attachment of monocytes to vascular endothelial cells and thus alleviates atherosclerosis. This study looks at whether GPR39 protects macrophages against ox-LDL-induced inflammation and apoptosis and ameliorates lipid accumulation in atherosclerosis and investigates its mechanism. Following inducement of ox-LDL, the expression of GPR39 and tumor necrosis factor alpha-induced protein 3 (TNFAIP3, also known as A20) in Raw 264.7 cells was detected by RT-qPCR and western blotting. The viability of macrophages treated with GPR39 agonist was detected by a cell counting kit 8 kit. GPR39 and A20 expression in ox-LDL-challenged macrophages was assayed by RT-qPCR and western blot with or without GPR30 agonist. After transfection of small interfering RNA (siRNA)-A20, the expression of pro-inflammatory cytokine tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-6 and anti-inflammatory cytokine IL-10 as well as NF-κB p65 and COX2 was detected. Lipid accumulation was observed through Oil Red O Staining. Total cholesterol (TC) and free cholesterol (FC) in macrophages were detected by commercial kits. Lastly, macrophage apoptosis was observed through TUNEL, and apoptosis-related proteins were detected by western blotting . Results indicated that decreased expression of GPR39 and A20 was observed in ox-LDL-induced macrophages. GPR39 agonist significantly increased A20 expression in ox-LDL-treated macrophages. Furthermore, A20 interference reversed the inhibitory effect of GPR39 agonist on ox-LDL-induced inflammation, lipid accumulation, TC and FC overexpression as well as cell apoptosis. In conclusion, activating GPR39 alleviates ox-LDL-induced macrophage inflammation, lipid accumulation and apoptosis in an A20-dependent manner.

CTRP6 inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration.

Vascular smooth muscle cell (VSMC) proliferation and migration play critical roles in the development and progression of atherosclerosis. C1q/tumor necrosis factor-related protein 6 (CTRP6), a member of CTRPs family, was involved in cardiovascular diseases, inflammatory reaction and adipogenesis. However, the role of CTRP6 in VSMCs remains largely unknown. The purpose of this study is to investigate the effects of CTRP6 on VSMC proliferation and migration and explore the possible mechanism. Our results indicated that CTRP6 expression was dramatically down-regulated in human atherosclerotic tissues and in cultured VSMCs stimulated by platelet-derived growth factor-BB (PDGF-BB). In addition, CTRP6 overexpression significantly inhibited the proliferation and migration of VSMCs exposed to PDGF-BB, as well as increased expression of α-SMA and SM22α in PDGF-BB-stimulated VSMCs. Furthermore, CTRP6 overexpression efficiently prevented the activation of PI3K/Akt/mTOR in VSMCs in response to PDGF-BB. In conclusion, these findings showed that CTRP6 inhibits PDGF-BB-induced VSMC proliferation and migration, at least in part, through suppressing the PI3K/Akt/mTOR signaling pathway. Therefore, CTRP6 may be a potential target for the treatment of atherosclerosis.

Prospective Randomized Study of Sarpogrelate Versus Clopidogrel-based Dual Antiplatelet Therapies in Patients Undergoing Femoropopliteal Arterial Endovascular Interventions: Preliminary Results.

BACKGROUND: Sarpogrelate is a selective 5-hydroxytryptamine (5-HT) receptor subtype 2A antagonist which blocks 5-HT induced platelet aggregation and proliferation of vascular smooth muscle cells. We compared the efficacy of sarpogrelate-based dual antiplatelet therapies for the prevention of restenosis and target lesion revascularization (TLR) rates comparing with that of clopidogrel after percutaneous endovascular interventions (EVIs) of femoropopliteal (FP) arterial lesions. METHODS: This prospective, multicenter, randomized clinical trial recruited a total of 120 patients with successful EVI of FP lesions at seven centers across China between January 2011 and June 2012. Patients were randomized to receive either sarpogrelate (100 mg trice daily for 6 months, n = 63) or clopidogrel (75 mg once daily for 6 months, n = 57). All patients also received oral aspirin (100 mg once daily for 12 months). Clinical follow-up was conducted up to 12 months postprocedure. RESULTS: There was no significant difference between the two groups in basic demographic data. The restenosis rate was higher in the clopidogrel group (22.80%) than in sarpogrelate group (17.50%), but there was no significant difference between these two groups (P = 0.465). The TLR rate, ipsilateral amputation rate, mortality in all-cause and bleeding rate were also similar in the two groups (P > 0.05). CONCLUSIONS: Aspirin plus sarpogrelate is a comparable antithrombotic regimen to aspirin plus clopidogrel after EVI of FP arterial lesions. Dual antiplatelet therapies might play an important role in preventing restenosis after successful EVI of FP lesions.

Characteristics of immunogenic and tolerogenic dendritic cells within the arterial wall in atherosclerosis and in vitro.

AIM: To investigate the characteristic of mature dendritic cell (mDC) and tolerogenic dendritic cell (TDC) in human lower limb atherosclerosis occlusion syndrome (ASO) and diabetic foot and in vitro. METHODS: 58 human ASO and diabetic foot arterial specimens were collected from surgical operation and autopsy. Immunohistochemical and Western blotting method were used to examine the distribution and the content of CD83 and CD1a positive reaction mDC and CD11b and DC-SIGN positive reaction TDC. Furthermore, bone marrow-derived DCs were induced by rmGM-CSF and rmIL-4 in the presence or absence of LPS in vitro. The percent of CD11c(+)CD11b(+)TDC and CD11c(+)CD83(+)mDC were analyzed by flow cytometry. The effects of TDC and mDC on T lymphocytes were analyzed by the IL-17 level, the percent of Th17, and IL-17 mRNA expression. RESULTS: Immunogenicity mDC was heavily found in intima plaque and around the small vessel of adventitia on artherosclerosis aorta of lesion group, and was positively correlated to the progress of the disease. However, there were low expression of TDC and was negatively correlated to the progress of the disease. Meanwhile, we found that there is a close relationship between high glucose and disease progression. TDC expressed high levels of IL-10 and TGF-ß1 and down-regulated the percent of CD4(+)IL-17(+) Th17, IL-17 mRNA and the level of IL-17 in vitro. CONCLUSION: TDC and mDC are assembled in the process of ASO, and the progression of the disease might be aggravated by DC-maturation. High glucose might closely relate to the progression of atherosclerosis.

Expression of tissue factor pathway inhibitor 2 in human pancreatic carcinoma and its effect on tumor growth, invasion, and migration in vitro and in vivo.

Tissue factor pathway inhibitor 2 (TFPI-2), also known as placental protein and matrix-associated serine protease inhibitor, plays an important role in angiogenesis, intravascular fibrinolysis, wound healing, tumor invasion, metastasis by plasmin, and trypsin mediated activation of zymogen matrix metalloproteinases. To detect whether TFPI-2 can be expressed in human pancreatic carcinoma samples and to investigate its role in the growth, invasion, and metastasis of pancreatic carcinoma cell in vitro and in vivo, we collected eight normal pancreatic tissue samples and 50 pancreatic carcinoma samples, and stably transfected the human pancreatic carcinoma cell line Panc-1 with a vector capable of expressing TFPI-2 gene. RT-PCR and Western blot analysis revealed that the levels of TFPI-2 expression were markedly lower in pancreatic carcinoma samples compared with normal pancreas samples. The level of TFPI-2 protein was significantly higher in cells transfected with TFPI-2 gene than that in the untransfected cells. The results of MTT assay showed that TFPI-2 inhibited Panc-1 cells growth in vitro. The invasive capacity of the cells transfected with TFPI-2 gene was also markedly less than that of untransfected cells in vitro as determined by the Matrigel invasion/migration assay. Moreover, TFPI-2 inhibited tumor growth, invasion, and metastasis in vivo in an orthotopic pancreatic carcinoma model. Our findings suggest that TFPI-2 plays a significant role in the growth, invasion, and metastasis of pancreatic carcinoma cell in vitro and in vivo, and has potential in anticancer therapy.

Prognostic significance of tissue factor pathway inhibitor-2 in pancreatic carcinoma and its effect on tumor invasion and metastasis.

Tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated kunitz-type serine proteinase inhibitor that plays an important role in plasmin and trypsin-mediated activation of zymogen matrix metalloproteinases involved in tumor angiogenesis, invasion and metastasis. Earlier studies have shown that the production of TFPI-2 is downregulated during the progression of various tumors. To detect whether TFPI-2 can be expressed in human pancreatic carcinoma samples, to evaluate its prognostic significance on pancreatic carcinoma and to investigate its effect on tumor invasion and metastasis, we collected 9 normal pancreatic tissue samples and 41 pancreatic carcinoma samples and stably transfected the human pancreatic carcinoma cell line Panc-1 with a vector capable of expressing TFPI-2 gene. RT-PCR and Western blot analysis revealed that the expression of TFPI-2 in pancreatic carcinoma samples was markedly lower than that in normal pancreas samples, and there was no TFPI-2 expression in Panc-1 cell. Its expression was related with biological characters of pancreatic carcinoma. The results of Boyden chamber assay and orthotopic pancreatic carcinoma model showed that TFPI-2 could inhibit invasion and metastasis ability of pancreatic carcinoma in vitro and in vivo. Kaplan-Meier survival curve and Cox proportional hazards model assay identified TFPI-2 as an independent prognostic factor for pancreatic carcinoma. Our data suggest that TFPI-2 plays a significant role in the invasion and metastasis of pancreatic carcinoma cell in vitro and in vivo and is determined to be an important prognostic factor for pancreatic carcinoma patients.

[Clinical significance of anticoagulant proteins detection in patients with thrombotic events].

OBJECTIVE: To investigate the prevalence and the risk of natural anticoagulants such as plasma protein C (PC), protein S (PS) and antithrombin (AT) deficiency in thromboembolic patients with no evident acquired factors. METHODS: Clotting assays on French STAGO autoanalyzer were used to detect the activity of plasma PC, PS and AT in 85 patients with thrombotic disease and 50 sex and age matched healthy controls. RESULTS: Among the 85 enrolled patients (18 arterial and 67 venous thromboembolism), male to female ratio was 1.4 and the median age was 42 years (17-69). The activity of plasma PC, PS and AT in the pre-therapy thrombotic disease group, the thrombo-recurrence group, and the age < or = 45 years group were significantly lower than that is the healthy control group, the first thrombotic episodes group and the age > 45 years group respectively (P < 0.001, P < 0.01, P < 0.01). The overall deficiency rate of these three natural anticoagulants was 30.6%, PS deficiency was the commonest (10.6%), the second was PC deficiency (8.2%), AT deficiency and combined deficiency each accounted for 5.9%. CONCLUSION: The PC, PS and AT protein deficiencies are frequent in Chinese thromboembolic patients, they are the independent risk factors for the thrombotic events and recurrence.

[Expression of TFPI-2 gene in pancreatic carcinoma and its prognostic significance].

OBJECTIVE: To detect the expression of TFPI-2 gene in pancreatic carcinoma, and to evaluate its prognostic significance in patients with pancreatic carcinoma. METHODS: Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of TFPI-2 mRNA and protein in the pancreatic carcinoma tissue samples from 41 patients. The correlation of its expression with clinicopathological features and its prognostic significance were analyzed. RESULTS: The expression of TFPI-2 mRNA and protein in moderately or poorly differentiated pancreatic carcinoma tissues, or in cases with nerve-involvement, lymph node and blood vessel invasion was significantly lower than that in the highly differentiated one, or without nerve involvement, or neither lymphatic nor blood vessel invasion (P < 0.05). The median survival time of patients with high expression of TFPI-2 (12.0 months) was significantly longer than that with low expression of TFPI-2 (5.1 months, P < 0.05). The Cox model analysis showed that the expression of TFPI-2 was an independent marker for prognosis in patients with pancreatic carcinoma. CONCLUSION: The expression of TFPI-2 is correlated with clinical stage and differentiation of pancreatic carcinoma, and can be used as a prognostic marker for pancreatic carcinoma.

Partial portacaval shunt with H-grafts to treat portal hypertension.

Partial portosystemic shunts have been popularized because of a reported low rate of mortality and morbidity (especially encephalopathy, liver failure and occlusion). The results of partial portacaval shunts [small-diameter expanded polytetrafluoroethylene (ePTFE) H-graft portacaval shunt] were retrospectively reviewed to evaluate the clinical efficacy in the treatment of portal hypertension. Forty-three patients with portal hypertension were treated by small-diameter H-graft of ePTFE portacaval shunt from May 1995 to April 2006. Thirty-three had externally ringed grafts and ten had non-ringed ones. Ten had grafts of 10 mm in diameter and 33 had grafts of 8 mm. The left gastric artery and coronary vein were ligated in all the cases. Six had pericardial devascularization and splenectomy was performed in 42. An average decrease of free portal pressure (FPP) from (33.24 ± 4.78) cmH2O before shunting and (13.65 ± 5.65) cmH2O after shunting was observed. The portal blood flow was reduced by one-third of that before shunt. Thirty-eight patients survived and no upper gastro-intestinal rebleeding occurred in the follow-up period (50.5 months in average). Two were out of contact. Color Doppler ultrasonography and/or portography revealed the shunts were patent in 38 cases and were occluded in three cases (3/41, 7.3%). Encephalopathy developed in five cases (5/41, 12.2%). Partial (small-diameter ePTFE H-graft) portacaval shunting can reduce the portal pressure effectively. Majority of the hepatic flow from the portal vein can be maintained adequately. The shunts with reinforced grafts can keep a higher rate of patency. The morbidity of encephalopathy was lower than those with total shunt. The partial portacaval shunt is effective in preventing recurrent variceal bleeding.

Small-diameter prosthetic H-graft portacaval shunts in the treatment of portal hypertension.

BACKGROUND: Portasystemic shunts, especially total shunts, are effective tools for reducing portal pressure and controlling variceal bleeding but lead to high risk of encephalopathy and accelerating liver failure. The purpose of this study is to evaluate the clinical effects of small-diameter expanded polytetrafluoroethylene (ePTFE) H-graft portacaval shunts in the treatment of portal hypertension. METHODS: Thirty-one patients with portal hypertension were treated with ePTFE small-diameter H-graft portacaval shunts from December 1995 to April 2002. Twenty-one had externally ringed grafts and 10 had non-ringed grafts; 20 had 10 mm diameter grafts and 11 had 8 mm grafts. The left gastric artery and coronary vein were ligated in 22 patients. Additionally, 6 patients underwent pericardial devascularization, and splenectomies were performed on 30 patients. RESULTS: An average decrease of free portal pressure (FPP) from (32.13 +/- 4.86) cmH2O before shunting to (12.55 +/- 5.57) cmH2O after shunting was observed. Portal blood flow was reduced by 1/3 compared with the levels measured before shunting. Twenty-eight patients survived after the operation, and no upper gastrointestinal rebleeding occurred in the follow-up period (40.2 months on average). We lost contact with one patient. Color Doppler ultrasonography and/or portography revealed the shunts to be patent in 28 cases and occluded in 2 (6.4%) cases. Encephalopathy developed in 4 patients (12.9%). CONCLUSION: Small-diameter ePTFE H-graft portacaval shunts can effectively reduce portal pressure. Moreover, the majority of the hepatopetal flow from the portal vein can be adequately maintained. The reinforced shunts may achieve a higher rate of patency. Morbidity from encephalopathy was less frequent than in patients receiving total shunts. Small-diameter H-graft portacaval shunts are also effective in preventing recurrent variceal bleeding.