[Effects and Mechanisms of Homoharringtonine on Expression of CEBPA Protein].

OBJECTIVE: To investigate the effect of homoharringtonine (HHT) on CEBPA protein and explore the mechanism of HHT in the treatment of acute myeloid leukemia (AML) with double CEBPA mutations. METHODS: The K562 cell line expressing CEBPA p30 (K562 CEBPA p30) was established. Western blot was used to determine the changes of the expression of CEBPA protein in K562 CEBPA p30, U937 and MOLM-13 cell lines before and after treatments with HHT, daunorubicin (DNR) or cytarabine (Ara-C). The effects of protease inhibitors and protein synthesis inhibitors on the expression of CEBPA protein were also determined. RNA-seq was used to analyze the difference of gene expressions and pathway enrichments between HHT group and DNR group. RESULTS: Both the endogenous CEBPA protein in U937 and MOLM-13 cell lines and the exogenous CEBPA protein in K562 CEBPA p30 were decreased by HHT (P<0.05) while were not by DNR or Ara-C. Proteasome inhibitors can increase the expression of CEBPA protein (P<0.05) while protein synthesis inhibitors can decrease the expression of CEBPA protein (P<0.05). The ribosome biogenesis related pathways in K562 CEBPA p30 were upregulated in HHT group while were not in DNR group. CONCLUSION: HHT can inhibit the synthesis of CEBPA and reduce the expression of CEBPA protein and this may be the mechanism of HHT in the treatment of CEBPA-double-mutant AML.

Comprehensive transcriptomics and metabolomics analyses reveal that hyperhomocysteinemia is a high risk factor for coronary artery disease in a chinese obese population aged 40-65: a prospective cross-sectional study.

BACKGROUND: Clinical observations suggest a complex relationship between obesity and coronary artery disease (CAD). This study aimed to characterize the intermediate metabolism phenotypes among obese patients with CAD and without CAD. METHODS: Sixty-two participants who consecutively underwent coronary angiography were enrolled in the discovery cohort. Transcriptional and untargeted metabolomics analyses were carried out to screen for key molecular changes between obese patients with CAD (CAD obese), without CAD (Non-CAD obese), and Non-CAD leans. A targeted GC-MS metabolomics approach was used to further identify differentially expressed metabolites in the validation cohorts. Regression and receiver operator curve analysis were performed to validate the risk model. RESULTS: We found common aberrantly expressed pathways both at the transcriptional and metabolomics levels. These pathways included cysteine and methionine metabolism and arginine and proline metabolism. Untargeted metabolomics revealed that S-adenosylhomocysteine (SAH), 3-hydroxybenzoic acid, 2-hydroxyhippuric acid, nicotinuric acid, and 2-arachidonoyl glycerol were significantly elevated in the CAD obese group compared to the other two groups. In the validation study, targeted cysteine and methionine metabolomics analyses showed that homocysteine (Hcy), SAH, and choline were significantly increased in the CAD obese group compared with the Non-CAD obese group, while betaine, 5-methylpropanedioic acid, S-adenosylmethionine, 4-PA, and vitamin B2 (VB2) showed no significant differences. Multivariate analyses showed that Hcy was an independent predictor of obesity with CAD (hazard ratio 1.7; 95%CI 1.2-2.6). The area under the curve based on the Hcy metabolomic (HCY-Mtb) index was 0.819, and up to 0.877 for the HCY-Mtb.index plus clinical variables. CONCLUSION: This is the first study to propose that obesity with hyperhomocysteinemia is a useful intermediate metabolism phenotype that could be used to identify obese patients at high risk for developing CAD.

C-reactive protein derived from perivascular adipose tissue accelerates injury-induced neointimal hyperplasia.

AIM: Inflammation within the perivascular adipose tissue (PVAT) in obesity plays an important role in cardiovascular disorders. C-reactive protein (CRP) level in obesity patients is significantly increased and associated with the occurrence and progression of cardiovascular disease. We tested the hypothesis CRP derived from PVAT in obesity contributes to vascular remodeling after injury. METHODS: A high-fat diet (HFD) significantly increased CRP expression in PVAT. We transplanted thoracic aortic PVAT from wild-type (WT) or transgenic CRP-expressing (CRPTG) mice to the injured femoral artery in WT mice. RESULTS: At 4 weeks after femoral artery injury, the neointimal/media ratio was increased significantly in WT mice that received PVAT from CRPTG mice compared with that in WT mice that received WT PVAT. Transplanted CRPTG PVAT also significantly accelerated adventitial macrophage infiltration and vasa vasorum proliferation. It was revealed greater macrophage infiltration in CRPTG adipose tissue than in WT adipose tissue and CRP significantly increased the adhesion rate of monocytes through receptor Fcγ RI. Proteome profiling showed CRP over-expression promoted the expression of chemokine (C-X-C motif) ligand 7 (CXCL7) in adipose tissue, transwell assay showed CRP increased monocyte migration indirectly via the induction of CXCL7 expression in adipocytes. CONCLUSION: CRP derived from PVAT was significantly increased in HFD mice and promoted neointimal hyperplasia after vascular injury.

Endoplasmic reticulum stress in perivascular adipose tissue promotes destabilization of atherosclerotic plaque by regulating GM-CSF paracrine.

BACKGROUND: Perivascular adipose tissue (PVAT) accelerates plaque progression and increases cardiovascular risk. We tested the hypothesis that PVAT contributed to plaque vulnerability and investigated whether endoplasmic reticulum stress (ER stress) in PVAT played an important role in vulnerable plaque. METHODS: We transplanted thoracic aortic PVAT or subcutaneous adipose tissue as a control, from donor mice to carotid arteries of recipient apolipoprotein E deficient (apoE-/-) mice after removing carotid artery collar placed for 6 weeks. Two weeks after transplantation, ER stress inhibitor 4-phenyl butyric acid (4-PBA) was locally administrated to the transplanted PVAT and then animals were euthanized after 4 weeks. Immunohistochemistry was performed to quantify plaque composition and neovascularization. Mouse angiogenesis antibody array kit was used to test the angiogenic factors produced by transplanted adipose tissue. In vitro tube formation assay, scratch wound migration assay and mouse aortic ring assay were used to assess the angiogenic capacity of supernatant of transplanted PVAT. RESULTS: Ultrastructural detection by transmission electron microscopy showed transplanted PVAT was a mixed population of white and brown adipocytes with abundant mitochondria. Transplanted PVAT increased the intraplaque macrophage infiltration, lipid core, intimal and vasa vasorum neovascularization and MMP2/9 expression in plaque while decreased smooth muscle cells and collagen in atherosclerotic plaque, which were restored by local 4-PBA-treatment. Antibody array analysis showed that 4-PBA reduced several angiogenic factors [Granulocyte Macrophage Colony Stimulating Factor (GM-CSF), MCP-1, IL-6] secreted by PVAT. Besides, conditioned medium from 4-PBA treated-PVAT inhibited tube formation and migration capacity of endothelial cells and ex vivo mouse aortic ring angiogenesis compared to conditioned medium from transplanted PVAT. mRNA expression and protein levels of GM-CSF were markedly elevated in adipocytes under ER stress which would be suppressed by 4-PBA. In addition, ER stress enhanced NF-κB binding to the promoter of the mouse GM-CSF gene in adipocytes confirmed by Chromatin immunoprecipitation analyses. CONCLUSIONS: Our findings demonstrate that ER stress in PVAT destabilizes atherosclerotic plaque, in part through increasing GM-CSF paracrine via transcription factor NF-κB.

Macrophage-derived foam cells impair endothelial barrier function by inducing endothelial-mesenchymal transition via CCL-4.

Recently, endothelial-mesenchymal transition (EndMT) has been demonstrated to play an important role in the development of atherosclerosis, the molecular mechanisms of which remain unclear. In the present study, scanning electron microscopy directly revealed a widened endothelial space and immunohistofluorescence demonstrated that EndMT was increased in human aorta atherosclerotic plaques. M1 macrophage-derived foam cell (M1-FC) supernatants, but not M2 macrophage-derived foam cell (M2-FC) supernatants, induced EndMT. A protein array and enzyme-linked immunosorbent assay identified that the levels of several cytokines, including C-C motif chemokine ligand 4 (CCL-4) were increased in M1-FC supernatants, in which EndMT was promoted, accompanied by increased endothelial permeability and monocyte adhesion. Furthermore, anti-CCL-4 antibody abolished the effects of M1-FC supernatants on EndMT. At the same time, CCL-4 activated its receptor, C-C motif chemokine receptor-5 (CCR-5), and upregulated transforming growth factor-ß (TGF-ß) expression. Further experiments revealed that EndMT induced by CCL-4 was reversed by treatment with CCR-5 antagonist and the RNA-mediated knockdown of TGF-ß. On the whole, the data of the present study suggest that M1-FCs induce EndMT by upregulating CCL-4, and increase endothelial permeability and monocyte adhesion. These data may help to elucidate the important role of EndMT in the development of atherosclerosis.

Functional assessment of transplanted kidneys with magnetic resonance imaging.

Kidney transplantation has emerged as the treatment of choice for many patients with end-stage renal disease, which is a significant cause of morbidity and mortality. Given the shortage of clinically available donor kidneys and the significant incidence of allograft dysfunction, a noninvasive and accurate assessment of the allograft renal function is critical for postoperative management. Prompt diagnosis of graft dysfunction facilitates clinical intervention of kidneys with salvageable function. New advances in magnetic resonance imaging (MRI) technology have enabled the calculation of various renal parameters that were previously not feasible to measure noninvasively. Diffusion-weighted imaging provides information on renal diffusion and perfusion simultaneously, with quantification by the apparent diffusion coefficient, the decrease of which reflects renal function impairment. Diffusion-tensor imaging accounts for the directionality of molecular motion and measures fractional anisotropy of the kidneys. Blood oxygen level-dependent MR evaluates intrarenal oxygen bioavailability, generating the parameter of R2* (reflecting the concentration of deoxyhemoglobin). A decrease in R2* could happen during acute rejection. MR nephro-urography/renography demonstrates structural data depicting urinary tract obstructions and functional data regarding the glomerular filtration and blood flow. MR angiography details the transplant vasculature and is particularly suitable for detecting vascular complications, with good correlation with digital subtraction angiography. Other functional MRI technologies, such as arterial spin labeling and MR spectroscopy, are showing additional promise. This review highlights MRI as a comprehensive modality to diagnose a variety of etiologies of graft dysfunction, including prerenal (e.g., renal vasculature), renal (intrinsic causes) and postrenal (e.g., obstruction of the collecting system) etiologies.

CHADS2 versus CHA2DS2-VASc score in assessing the stroke and thromboembolism risk stratification in patients with atrial fibrillation: a systematic review and meta-analysis.

OBJECTIVE: To perform a systematic review and meta-analysis of the predictive abilities of CHADS2 and CHA2DS2-VASc in stroke and thromboembolism risk stratification of atrial fibrillation (AF) patients. METHODS: We searched PubMed and EMBASE for English-language literature on comparisons of the diagnostic performance between CHADS2 and CHA2DS2-VASc in predicting stroke, or systemic embolism, in AF. We then assessed the quality of the included studies and pooled the C-statistics and 95% confidence intervals (95% CI). RESULTS: Eight studies were included. It was unsuitable to perform a direct meta-analysis because of high heterogeneity. When analyzed as a continuous variable, the C-statistic ranged from 0.60 to 0.80 (median 0.683) for CHADS2 and 0.64-0.79 (median 0.673) for CHA2DS2-VASc. When analyzed as a continuous variable in anticoagulation patients, the subgroup analysis showed that the pooled C-statistic (95% CI) was 0.660 (0.655-0.665) for CHADS2 and 0.667 (0.651-0.683) for CHA2DS2-VASc (no significant difference). For non-anticoagulation patients, the pooled C-statistic (95% CI) was 0.685 (0.666-0.705) for CHADS2 and 0.675 (0.656-0.694) for CHA2DS2-VASc (no significant difference). The average ratio of endpoint events in the low-risk group of CHA2DS2-VASc was less than CHADS2 (0.41% vs. 0.94%, P < 0.05). The average proportion of the moderate-risk group of CHA2DS2-VASc was lower than CHADS2 (11.12% vs. 30.75%, P < 0.05). CONCLUSIONS: The C-statistic suggests a similar clinical utility of the CHADS2 and CHA2DS2-VASc scores in predicting stroke and thromboembolism, but CHA2DS2- VASc has the important advantage of identifying extremely low-risk patients with atrial fibrillation, as well as classifying a lower proportion of patients as moderate risk.

[Chemical constituents of Desmodium caudatum].

OBJECTIVE: To study the chemical constituents of Desmodium caudatum. METHODS: Silica column chromatography, Sephadex LH-20 column chromatography and recrystallization were used to separate and purify the chemical composition of Desmodium caudatum. Their chemical structures were identified by infrared spectrum (IR), mass spectrum (MS), nuclear magnetic resonance (NMR) and other physicochemical methods. RESULTS: Twelve compounds were isolated and identified as lacceroic acid(1), gheddic acid(2), stigmasterol(3), betulin(4), citrusinol(5), yukovanol(6), kaempferol(7), protocatechuic acid(8), sophocarpine(9), matrine(10), N, Ndimethyltryptamine(11) and 5-hydroxy-N,N-dimethyltryptamine(12). CONCLUSION: Compounds 1, 2, 4 and 8-12 are isolated from this plant for the first time.

[The value of gastric bare area, left adrenal gland and perirenal space involvement in predicting complications of acute pancreatitis].

OBJECTIVE: To investigate CT findings and incidence rate of gastric bare area involvement (GBAI), left adrenal gland involvement (LAGI) and perirenal space involvement (PSI) in acute pancreatitis, and to also explore the value of these appearances in predicting complications and mortality of patients. METHODS: CT imaging data of 575 patients with AP diagnosed by clinic from October 2009 to April 2011 were analyzed retrospectively. There were 339 male and 236 female patients, aging from 16 to 93 years with a mean of (51 ± 16) years. Involvement with or without gastric bare area, left adrenal gland, perirenal space were focused, and the relationship were analyzed between these CT findings and complications and mortality of patients. RESULTS: Among 167 patients (29.0%) with GBAI, 132 had complications and 16 died. The sensitivity and specificity of GBAI for predicting complications were 45.4% and 87.7%, respectively, and 84.2%and 72.8% for predicting mortality. In all 107 patients (18.6%) with LAGI, 81 had complications and 18 died. The sensitivity and specificity of LAGI for predicting complications were 27.8% and 90.8%, respectively, and 94.7% and 84.0% for predicting mortality. Among 335 patients (58.3%) with PSI, 201 had complications and 19 died. The sensitivity and specificity of PSI for predicting complications were 69.1% and 52.8%, respectively, and 100% and 43.2% for predicting mortality. Of all patients, 210 (36.5%) owned two or more positive CT findings among GBAI, LAGI, and PSI. One hundred ninety-eight of these patients had complications and 19 died, that predicted the sensitivity and specificity for complications were 68.0% and 95.8%, respectively, and 100% and 65.6% for mortality. The risk of complications in the patients with GBAI or LAGI was increased than normal gastric bare area or left adrenal gland 5.9 or 3.8 times respectively, and the risk of death was improved 14.3 or 94.5 times respectively. The risk of complications in those patients with two or more positive findings increased by 48.3 times. By analyzing receiver operating characteristic (ROC) curve, the combination of GBAI, LAGI, and PSI was the best way to predict the complications of AP. The area under the ROC was 0.819. CONCLUSIONS: The CT imaging could effectively indicate the signs of GBAI, LAGI and PSI of AP. The GBAI. LAGI and PSI were related with the severity and prognosis of AP, and these findings could be clinical indicators for evaluating prognosis of AP.

Oncogenic HPV infection resembling pseudocondyloma of vulvae: a descriptive study of 31 cases.

OBJECTIVE: Report a special type of human papillomavirus infection resembling pseudocondyloma of vulvae. The characteristics of the clinical manifestation and subtypes of the HPV infection were studied. METHODS: Thirty-one women with unprotected sex and genital warts resembling pseudocondyloma of vulvae were involved in this study. The clinical and histological manifestations were characterized. The genotypes of human papillomavirus were determined by a reverse hybridization of polymerase chain reaction (PCR) RESULTS: Thirteen HPV genotypes were detected from 31 cases. Of 13 genotypes, 3 were non-oncogenic and 10 were oncogenic. At least, one of oncogenic genotypes was detected from each patient. Among 31 cases, 10 were found with one genotype of the HPV infection, 14 with 2 genotypes of HPV and 7 with at least 3 genotypes. The most common genotype was genotype 16 that was detected from 15 of 31 samples (48%). CONCLUSION: There is a special type of oncogenic HPV infection clinically resembling pseudocondyloma of vulvae. Some of the patients who were infected with this type of HPV infection may have been misdiagnosed as pseudocondyloma of vulvae in the past time.

[Comparison of coronary angiography with myocardial perfusion imaging in assessment of functionally relevant coronary artery lesion].

OBJECTIVE: To evaluate the accuracy of dual-source CT coronary angiography (DSCTCA) for the depiction of functionally relevant coronary artery lesion(FRCAL), by using myocardial perfusion imaging (MPI) with single photon emission computed tomography (SPECT). METHODS: DSCTCA, (99)Tc(m)-MIBI SPECT myocardial perfusion imaging (MPI) and conventional coronary angiography (CCA) were performed in 59 patients with clinical suspected CAD. Coronary artery diameter narrowing of 50% or greater at DSCTCA was defined as stenosis and was compared with MPI findings. CCA was served as a reference standard for DSCTCA. RESULTS: (1) Agreement between DSCTCA and CCA was good (kappa = 0.93 for patient-based analysis, Kappa = 0.88 for vessel-based analysis). (2) DSCTCA revealed stenoses in 86 segments corresponding to 60 arteries in 34 patients. (3) MPI revealed 19 reversible, 21 partially reversible, and 5 fixed defects in 25 patients. (4) About 65.0% (39/60) of all the narrowed coronary arteries were determined to be FRCAL. Sensitivity, specificity, accuracy, positive predictive values and negative predictive values, respectively, of DSCTCA in the detection of all MPI defects were 92.0%, 67.6%, 78.0%, 67.6% and 92.0% on a per-patient basis and 86.7%, 89.0%, 88.6%, 65.0% and 96.6% on a per-artery basis. (5) ROC analysis showed that predictive value of DSCTCA in FRCAL was similar with those of CCA (AUCs = 0.80, 0.82). CONCLUSIONS: DSCTCA can evaluate FRCAL indirectly. When DSCTCA results are negative, it can help ruled out patients with FRCAL. The positive DSCTCA results should combine MPI in predictor of myocardial ischemia.

[Clinical applications of routine subtraction and dual-energy subtraction cervical arteries computed tomographic angiography for cervical arteries imaging].

OBJECTIVE: To compare with the clinical applications of routine-subtraction and dual-energy subtraction cervical arteries computed tomographic angiography (CTA) for cervical arteries imaging. METHODS: Scanning was performed in 45 patients with clinically suspected cervical arteries disease with dual-source CT. The data of two different energy were collected only at one scanning. The data post processing include: conventional bone-removal digital subtraction (routine-subtraction) was performed with plain and 80 kV enhanced scanning. Volume render (VR) and maximum intensity projection (MIp) reconstruction were finished. Direct bone-removal digital subtraction (dual-energy subtraction) was performed with 80 and 140kV enhanced scanning that have different energy, and saving the data of subtraction. VR and MIp reconstruction were finished. The image quality, which was divided into four grades, was compared between these two groups. The effective radiation dose was also compared. RESULTS: For normal vessels, no abnormality was found in 24 of 45 cases, with the common carotid artery and its branches clearly displayed with both two methods. The image quality was not significantly different between dual-energy subtraction CTA and routine subtraction CTA (P>0.05) . For stenotic vessels, 45 stenotic vessels in 21 cases were clearly displayed clearly with both two methods (P>0.05) . The effective radiation dose was decreased by 17.3 % for dual-energy subtraction CTA when compared with routine-subtraction CTA (P<0.01) . CONCLUSIONS: Both routine-subtraction and dual-energy subtraction CTA can clearly display normal and stenotic vessels. The radiation exposure dose is relatively lower in dual-energy CTA. The dual-energy subtraction CTA has better effectiveness when used for non-cooperation patients.

[Studies on the chemical constituents from the antithrombus activity extract of Radix Aerio Fici Microcarpae].

OBJECTIVE: To study the chemical constituent from the antithrombus activity extract of Radix Aerio Fici Microcarpae. METHODS: The isolation and purification of the extract were conducted by means of silica gel column chromatography, polyamide column chromatography and recrytallization. The structures of these compounds were elucidated based on the physicochemical properties and spectroscopic analysis. RESULTS: 7 compounds were isolated from the extract. They were identified as isovanillic acid (1), 4-hydroxybenzoic acid(2),3,4-dihydroxybenzoic acid (3), friedelane (4), alpha-amyrin acetate (5), lanosterol (6) and beta-sitosterol (7) . CONCLUSION: Compound 3,4 and 7 are isolated from this plant for the first time.

[The value of sixty-four slice spiral CT angiography in diagnosing and evaluating abdominal aortic aneurysm before operations].

OBJECTIVE: To assess the value of sixty-four slice spiral CT angiography (CTA) in diagnosing and evaluating abdominal aortic aneurysm (AAA) before operations. METHODS: Between Oct 2006 and Jan 2008, forty-three consecutive patients with suspected AAA underwent examinations by the sixty-four slice spiral CT angiography with 370 mg I/mL contrast material. Of these patients, 38 patients whose AAA imaging quality met the diagnostic criteria were enrolled in the study. Imaging reconstruction were made at the workstation, which included multi-plane reconstruction (MPR), maximum intensity projection (MIP) and volume rendering technique (VRT). The comprehensive observation and analysis focused on the type, location, size and extension of AAA. The classification of AAA was based on the relevant parameters that satisfied the requirement of endovascular stent-graft exclusion. RESULTS: (1) The aneurysms of all of the 38 patients were located at under the level of renal arteries, including six that were adjacent to renal arteries and 32 under renal arteries. (2) Among the 38 patients with AAA, 4 were type I , 3 were type II A, 4 were type II B, 21 were type II C, and 6 were type III. (3) The sixty-four slice spiral CT angiography clearly displayed the shape, locality and size of the aneurysms, the anatomic relations between the aneurismal bodies and the adjacent main branches of aorta, and the mural thrombus in all of the patients. Relevant parameters of AAA could be accurately measured by the CTA images. CONCLUSION: Sixty-four slice spiral CT angiography is a fast, non-invasive and effective instrument for diagnosing and evaluating AAA before operations.

[A new flavonol glycoside from Baeckea Frutescens L].

To study the chemical constituents of the traditional Chinese herb Baeckea Frutescens L., a new flavonol glycoside, named 6, 8-dimethylkaempferol-3-O-alpha-L-rhamnoside (1), together with seven known compounds: quercetin (2), quercetin-3-O-alpha-L-rhamnoside (3), myricetin (4), myricetin-3-O-alpha-L-rhamnoside (5), gallic acid (6), ursolic acid (7) and 1,3-dihydroxy-2-(2'-methoxylpropionyl)-5-methoxy-6-methylbenzene (8) were isolated by using silica gel column chromatography, polyamide column chromatography and recrytallization. Their structures were identified on the basis of physicochemical properties and spectroscopic analysis. Among them, compounds 2-7 were isolated from this plant for the first time and compound 8 was first isolated from plant.