PRMT1 enhances oncogenic arginine methylation of NONO in colorectal cancer.

Arginine methylation is an important posttranslational modification catalyzed by protein arginine methyltransferases (PRMTs). However, the role of PRMTs in colorectal cancer (CRC) progression is not well understood. Here we report that non-POU domain-containing octamer-binding protein (NONO) is overexpressed in CRC tissue and is a potential marker for poor prognosis in CRC patients. NONO silencing resulted in decreased proliferation, migration, and invasion of CRC cells, whereas overexpression had the opposite effect. In a xenograft model, tumors derived from NONO-deficient CRC cells were smaller than those derived from wild-type (WT) cells, and PRMT1 inhibition blocked CRC xenograft progression. A mass spectrometry analysis indicated that NONO is a substrate of PRMT1. R251 of NONO was asymmetrically dimethylated by PRMT1 in vitro and in vivo. Compared to NONO WT cells, NONO R251K mutant-expressing CRC cells showed reduced proliferation, migration, and invasion, and PRMT1 knockdown or pharmacological inhibition abrogated the malignant phenotype associated with NONO asymmetric dimethylation in both KRAS WT and mutant CRC cells. Compared to adjacent normal tissue, PRMT1 was highly expressed in the CRC zone in clinical specimens, which was correlated with poor overall survival in patients with locally advanced CRC. These results demonstrate that PRMT1-mediated methylation of NONO at R251 promotes CRC growth and metastasis, and suggest that PRMT1 inhibition may be an effective therapeutic strategy for CRC treatment regardless of KRAS mutation status.

Generation of functional hepatocyte-like cells from human bone marrow mesenchymal stem cells by overexpression of transcription factor HNF4α and FOXA2.

BACKGROUND: Our previous study showed that overexpression of hepatocyte nuclear factor 4α (HNF4α) could directly promote mesenchymal stem cells (MSCs) to differentiate into hepatocyte-like cells. However, the efficiency of hepatic differentiation remains low. The purpose of our study was to establish an MSC cell line that overexpressed HNF4α and FOXA2 genes to obtain an increased hepatic differentiation efficiency and hepatocyte-like cells with more mature hepatocyte functions. METHODS: Successful establishment of high-level HNF4α and FOXA2 co-overexpression in human induced hepatocyte-like cells (hiHep cells) was verified by flow cytometry, immunofluorescence and RT-PCR. Measurements of albumin (ALB), urea, glucose, indocyanine green (ICG) uptake and release, cytochrome P450 (CYP) activity and gene expression were used to analyze mature hepatic functions of hiHep cells. RESULTS: hiHep cells efficiently express HNF4α and FOXA2 genes and proteins, exhibit typical epithelial morphology and acquire mature hepatocyte-like cell functions, including ALB secretion, urea production, ICG uptake and release, and glycogen storage. hiHep cells can be activated by CYP inducers. The percentage of both ALB and α-1-antitrypsin (AAT)-positive cells was approximately 72.6%. The expression levels of hepatocyte-specific genes (ALB, AAT, and CYP1A1) and liver drug transport-related genes (ABCB1, ABCG2, and SLC22A18) in hiHep cells were significantly higher than those in MSCs-Vector cells. The hiHep cells did not form tumors after subcutaneous xenograft in BALB/c nude mice after 2 months. CONCLUSION: This study provides an accessible, feasible and efficient strategy to generate hiHep cells from MSCs.

Nomogram for predicting disease-free survival among a multicenter cohort of Chinese patients with locally advanced rectal cancer.

Purpose: This study aimed to develop and validate a nomogram for predicting 3-year disease-free survival (DFS) among a multicenter cohort of Chinese patients with locally advanced rectal cancer (LARC) who underwent preoperative therapy followed by surgery. This nomogram might help identify patients who would benefit from postoperative adjuvant chemotherapy and close follow-up. Materials and methods: All data from 228 patients in two independent Chinese cohorts (118 patients and 110 patients) were pooled and subjected to survival analysis. One cohort's data were used to develop multivariate nomograms based on Cox regression, and the second cohort was used for external validation. The variables were sex, age, clinical tumor stage, tumor location, preoperative therapy protocol, adjuvant chemotherapy, surgical procedure, surgical approach, pTNM stage, tumor deposit, tumor regression grade, lymphovascular invasion, perineural invasion, pretreatment serum carcinoembryonic antigen (CEA) level, preoperative CEA level, and postoperative CEA level. The model's performance was evaluated based on its discrimination, calibration, and clinical usefulness. Results: The nomogram was based on ypT stage and ypN stage, and the C-index values for 3-year DFS were 0.70 in the training cohort (95% confidence interval: 0.62-0.78) and 0.78 in the validation cohort (95% confidence interval: 0.68-0.89). The Hosmer-Lemeshow calibration test revealed good calibration for predicting 3-year DFS in the training and validation cohorts, and decision curve analysis demonstrated that the nomogram was clinically useful. Conclusion: This nomogram including the ypT stage and ypN stage could predict DFS at 3 years after surgery, which may help better identify Chinese patients who would benefit from additional postoperative adjuvant systemic treatment.

Role of multi-detector computed tomography for biliary complications after liver transplantation.

AIM: To investigate the diagnostic performance of multi-detector computed tomography (MDCT) in detecting biliary complications after orthotopic liver transplantation (OLT). METHODS: Eighty-three consecutive OLT recipients, who presented with clinical or biochemical signs of biliary complications, underwent MDCT examination. Two experienced radiologists assessed MDCT images in consensus to determine biliary complications. Final confirmation was based on percutaneous transhepatic cholangiography or endoscopic retrograde cholangiography in 58 patients, surgery in four patients, liver biopsy in 10, and clinical and sonography follow-up in 11 patients. RESULTS: Biliary complications were eventually confirmed in 62 of 83 patients (74.7%), including anastomotic biliary strictures in 32, nonanastomotic biliary strictures in 21, biliary stones in nine (5 with biliary strictures), anastomotic bile leak in five, and biloma in six (all with nonanastomotic strictures, and 2 with biligenic hepatic abscess). Twenty-one patients had no detection of biliary complications. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of MDCT for detecting biliary strictures were 90.6%, 86.7%, 89.2%, 92.3% and 83.9%, respectively. For detecting biliary stones, anastomotic bile leak and biloma, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of MDCT were all 100%. CONCLUSION: MDCT is a useful screening tool for detecting biliary complications after OLT.

Renal aspergillosis after liver transplantation: clinical and imaging manifestations in two cases.

Renal aspergillosis (RAsp) is a rare complication in liver transplant (LT) recipients. Here we report RAsp in two LT recipients. In both patients, RAsp occurred more than 90 d after allogenetic orthotropic LT, and all the clinical findings were unspecific. RAsp involved unilateral kidney in Case one and bilateral kidneys in Case two. Both computed tomography (CT) and magnetic resonance imaging (MRI) revealed renal abscesses, with progressively enhanced walls and separations and unenhanced alveolate areas after contrast agent administration. On unenhanced CT images they showed inhomogeneous hypo-attenuation. On fat-suppressed T2-weighted images (T2WIs), the walls and separations of the abscesses showed slightly low signal intensity and the central parts of the lesions showed slightly high signal intensity. Both on CT and MRI, there were some hints of renal infarction or chronic ischemia. Both cases were treated by radical nephrectomy followed by adjuvant antifungal treatment. They all recovered well.

[Evaluation of graft perfusion in patients with ischemic-type of biliary lesions after liver transplantation].

OBJECTIVE: To investigate the value of 320-rows CT perfusion (CTP) imaging in the study of hepatic hemodynamic characters in ischemic-type biliary lesions (ITBL) after liver transplantation. METHODS: A total of 11 ITBL patients received 320-slice CT angiography (CTA) and CTP after liver transplantation scheduled at 5-10 min away. Four patients underwent liver biopsy While 7 patients with normal liver after transplantation were selected as the control group. The parameters of hepatic artery perfusion (HAP), portal vein perfusion (PVP), total hepatic perfusion (TLP) and hepatic arterial perfusion index (HPI) were measured and compared for all patients. And the blood perfusion characters of liver with ITBL after transplantation were analyzed. RESULTS: (1) In 11 ITBL patients, 3 patients had no vascular complications on CTA, 1 with simple hepatic artery stenosis (HAS), 1 with HAS and arterioportal shunt (APS), 2 with HAS and portal vein stenosis/right hepatic vein stenosis (PVS/RHVS), 1 with simple APS, 2 with simple PVS and 1 with portal vein thrombosis and cavernous transformation of portal vein (PVT and CTPV). And 4/11 patients underwent liver biopsy, 2 in which confirmed mild acute rejection and 2 confirmed biliary obstruction associated with ascending biliary infection.(2) HAP of the ITBL and control groups were (66 ± 38) and (40 ± 8) ml×min(-1)·(100 ml)(-1), PVP (128 ± 35) and (163 ± 21) ml×min(-1)·(100 ml)(-1), TLP (194 ± 58) and (203 ± 19) ml×min(-1)·(100 ml)(-1), HPI 34% ± 14% and 21% ± 4% respectively. The differences in the value of HAP, PVP and HPI between the groups were statistically significant (P < 0.05) excluding TLP. CONCLUSION: Various liver perfusion abnormalities of ITBL may be evaluated objectively by CTP. ITBL might occurred when HAP and HPI increased with a decreased of PVP.

[Effects of rho-kinase inhibitor on cardiac hypertrophy of left ventricle in spontaneously hypertensive rats].

OBJECTIVE: To explore the effects of Rho-kinase inhibitor on cardiac hypertrophy of left ventricle in spontaneously hypertensive rats (SHR). METHODS: SHRs (n = 30) were randomly divided into 5 groups (n = 6 each): SHR control group, 5 mg/kg fasudil group (SHRL), 10 mg/kg fasudil group (SHRM), 20 mg/kg fasudil group (SHRH) and nifedipine group (XBDP, 10 mg/kg). Six male Wistar-Kyoto rats were selected as normal control group (WKY group). Systolic blood pressure (SBP) was measured before and after treatment every 2 weeks. The hypertrophic parameters of left ventricular weight index (LVWI) and cardiomyocyte transverse diameter (TDM) were measured. In addition, the expression levels of Rho kinase mRNA and protein in cardiomyocytes were observed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: The levels of LVWI and TDM in WKY group were significantly lower than those in SHR control group [(2.98 ± 0.05) vs (3.16 ± 0.08) mg/g, (18.18 ± 0.75) vs (21.32 ± 1.25) µm, P < 0.01]. After 8 weeks, the levels of LVWI and TDM in three fasudil groups were markedly lower than those in SHR control group [SHRL group: (3.12 ± 0.05) mg/g, SHRM group: (3.10 ± 0.07) mg/g, SHRH group: (3.08 ± 0.09) mg/g vs SHR control group: (3.16 ± 0.08) mg/g, SHRL group: (20.11 ± 1.15) µm, SHRM group: (19.63 ± 1.62) µm, SHRH group: (18.91 ± 1.05) µm vs SHR control group: (21.32 ± 1.25) µm, P < 0.05 or P < 0.01]. But the levels of LVWI and TDM were not different between SHR control and XBDP groups [(3.14 ± 0.08) mg/g,(21.42 ± 1.23) µm, P > 0.05]. Compared with SHR control group, the expression of Rho kinase mRNA and protein in cardiomyocytes significantly decreased in three fasudil groups [SHRL group mRNA: (0.45 ± 0.08), SHRM group mRNA: (0.37 ± 0.07), SHRH group mRNA: (0.32 ± 0.07) vs SHR control group mRNA: (0.63 ± 0.07), SHRL group protein: 0.78 ± 0.11), SHRM group protein: (0.73 ± 0.10), SHRH group protein: (0.68 ± 0.10) vs SHR control group protein: (0.90 ± 0.1), P < 0.05 or P < 0.01], but showed no obvious change in XBDP group [mRNA: (0.56 ± 0.07), protein: (0.85 ± 0.10), P > 0.05]. CONCLUSIONS: Rho kinase inhibitor may significantly down-regulate the expression of Rho kinase in cardiomyocytes of SHR. The mechanism is probably due to the favorable effects of Rho kinase inhibitor in the prevention of cardiac hypertrophy of left ventricle.

[Clinical value of dual-source CT in evaluating coronary artery disease].

OBJECTIVE: To analyze the clinical value of dual-source CT (DSCT) in the diagnosis of coronary artery disease. METHODS: Fifty-five patients with suspected coronary heart disease underwent both DSCT coronary angiography (DSCTCA) and selective coronary angiography (CAG) examination, and the diagnostic sensitivity, specificity and accuracy of the DSCTCA was evaluated. RESULTS: The sensitivity, specificity, positive and negative predictive value, and accuracy of DSCT in the diagnosis of coronary heart disease were 97.7%, 72.6%, 93.5%, 88.9% and 92.7% by the number of patients, respectively; by calculating the coronary arteries, the sensitivity, specificity, positive and negative predictive value, accuracy were 94.9%, 95.8%, 92.5%, 97.1%, 95.5%, respectively. According to the lesion segment, these values were 88.2%, 96.9%, 90.5%, 96.1%, 94.7%, respectively. DSCTCA showed no significant difference from CAG for a diagnostic purpose, nor did their vessel sensitivity, specificity, positive and negative predictive value, and accuracy in different coronaries differ significantly. CONCLUSION: DSCT has a diagnostic accuracy of coronary heart disease close to that CAG and can on some occasion serve as an alternative to CAG in the screening of coronary artery disease.

Gossypol inhibits phosphorylation of Bcl-2 in human leukemia HL-60 cells.

Gossypol is an attractive therapeutic anti-tumor agent as an apoptosis inducer and is being evaluated in preclinical tests. However, the molecular mechanisms underlying apoptosis induction by gossypol in malignant cells have not been completely enunciated. Here we investigate the alterations of Bcl-2/Bcl-xL/Mcl-1 protein levels and Bcl-2 phosphorylation in gossypol-induced apoptosis in human leukemia HL-60 cells. We found that gossypol treatment inhibited cell growth and induced apoptosis in HL-60 cells. Bcl-2/Bcl-xL/Mcl-1 protein levels were slightly reduced and phosphorylation of Bcl-2 at threonine 56 (phospho T56) was not altered. However, phosphorylation of Bcl-2 at serine 70 (phospho S70) was strikingly down-regulated in gossypol-exposed cells. This reduction was found to be not only in both dose- and time-dependent fashion but also obviated by phorbol l2,13-dibutyrate (PDBu), an activator of protein kinase C (PKC). In addition, pre-treatment of PDBu partially prevented gossypol-induced apoptosis in HL-60 cells. Collectively, gossypol treatment can reduce phosphorylation of Bcl-2 at serine 70 in leukemia HL-60 cells and gossypol may be a promising therapeutical candidate for leukemia patients especially expressing phosphorylated Bcl-2 at Ser70.

A one-step immunotest for rapid detection of heart-type fatty acid-binding protein in patients with acute coronary syndromes.

Using heart-type fatty acid-binding protein (H-FABP) as an early cardiac marker for diagnosis of acute myocardial infarction (AMI) soon after the onset of symptoms requires a rapid assay. A one-step test called, CardioDetect, is used for detection of H-FABP in whole blood sample. Thirty patients suspected of AMI presenting to the emergency department within 12 hours after onset were enrolled in this study. The diagnostic performance of CardioDetect was compared with different cardiac markers. There were 59.1% of patients with positive H-FABP within 6 hours after onset, while there were only 18.2% with positive cardiac troponin I (cTnI). Results indicated the diagnostic power of H-FABP for AMI was significantly higher than that of cTnI. The sensitivity of H-FABP was 81.8%, which was higher than those of the other cardiac markers, while the specificity was comparable. The area under the receiver operating characteristic curve for H-FABP was 0.909, which was significantly larger than the others. With this rapid and sensitive immunotest, H-FABP could be soon introduced in clinical practice in combination with well-established markers like troponins.

[Correlation of adiponectin, monocyte chemoattractant protein-1, and endothelial function to vascular remodeling in coronary in-stent restenosis].

OBJECTIVE: To investigate the correlation between vascular remodeling index (RI) and serum adiponectin, plasma monocyte chemoattractant protein-1 (MCP-1), endothelial function and evaluate the mechanism of coronary in-stent restenosis. METHODS: RI 6 months after percutaneous coronary intervention (PCI), serum adiponectin, plasma MCP-1 and flow-mediated dilation (FMD) before and 3 days,6 months after PCI were measured in 30 patients with and 30 without coronary in-stent restenosis. RESULTS: Compared with patients without restenosis and those with restenosis before PCI, the patients with coronary in-stent restenosis showed significantly increased plasma MCP-1 3 days and 6 months after PCI (P<0.05) and reduced RI 6 months after PCI, serum adiponectin and FMD 3 days and 6 months after PCI (P<0.05). RI was positively correlated to serum adiponectin and FMD and inversely to MCP-1. CONCLUSION: The occurrence of coronary in-stent restenosis is the result of the interrelations between multiple factors.

[Effect of combination of Chinese and Western medicines on sinus rhythm maintenance in patients with auricular fibrillation after conversion].

OBJECTIVE: To investigate the curative effects of irbesartan, amiodarone and Wenxin Granule (WG), applied alone or in combination, on sinus rhythm maintenance in patients with auricular fibrillation (AF) after conversion. METHODS: Forty-one patients of persistent AF, after their fibrillation being converted, were divided into three groups randomly, and treated with amiodarone (group A, n=14), irbesartan and amiodarone (group B, n=15), and WG plus irbesartan and amiodarone (group C, n=12) respectively for 6 months. RESULTS: Compared with that before treatment, the inner diameter of atria sinistrum reduced in group B and C, and the reduction in the latter was superior to that in the former (P < 0.05); the diameter of left ventricle also reduced in group C (P < 0.05); and the maintenance rate of sinus rhythm was higher in group C than that in group A (P < 0.05). CONCLUSION: Combined therapy of Chinese and Western medicines shows synergistic effect of anti-arrhythmia.