Rationality and implication of catheter-based adrenal ablation for bilateral primary aldosteronism.

Mineralocorticoid receptor antagonists (MRAs) for bilateral primary aldosteronism (PA) are the mainstay option recommended by guidelines, but poor compliance occurs due to numerous side effects. We aimed to examine whether catheter-based adrenal ablation could be an alternative treatment for bilateral PA.644 PA patients were included from a total of 6054 hypertensive patients. Adrenal CT scan and adrenal venous sampling (AVS) were both performed for PA subtype classification. Clinical and biochemical outcomes were assessed at 6 months after treatment according to the Primary Aldosteronism Surgical Outcome (PASO) criteria.93 patients with PA were recruited to be treated by adrenal ablation, including 25 bilateral PA and 68 unilateral PA according to AVS results. Office SBP and DBP significantly decreased from baseline levels, serum potassium levels increased and ARR significantly decreased (p < 0.01) in both the bilateral and unilateral groups. In the bilateral group, complete, partial and absent clinical success was achieved in 6 (24.0%), 11 (44.0%) and 8(32.0%) patients, respectively. In the unilateral group, complete, partial and absent clinical success was achieved in 12 (17.6%), 37 (54.4%), and 19 (27.9%) patients, respectively. The numbers of patients achieving complete, partial, and absent biochemical success were 15 (60.0%), 6 (24.0%), and 4 (16.0%), respectively, in the bilateral group versus 37 (54.4%), 9 (13.2%), and 22 (32.3%), respectively, in the unilateral group. In conclusion, we provide evidence for the beneficial outcomes of unilateral adrenal ablation for patients with bilateral PA. Our findings provide insight into an alternative option for patients with bilateral excess aldosterone.

Machine Learning-Assisted Analysis of Sublingual Microcirculatory Dysfunction for Early Cardiovascular Risk Evaluation and Cardiovascular-Kidney-Metabolic Syndrome Stage in Patients With Type 2 Diabetes Mellitus.

AIMS: To examine whether sublingual microcirculation can be used as an effective and noninvasive method for assessing cardiovascular, kidney, and metabolic risks in patients with type 2 diabetes mellitus (T2DM). MATERIALS AND METHODS: This cross-sectional observational study enrolled 186 patients with T2DM. All patients were evaluated using the Framingham General Cardiovascular Risk Score (FGCRS) and cardiovascular-kidney-metabolic (CKM) syndrome stage. Side-stream dark-field microscopy was used for sublingual microcirculation, including total and perfused vessel density (TVD and PVD). Multiple machine-learning prediction models have been developed for CKM risk and stage assessment in T2DM patients. Receiver operating characteristic (ROC) curves were generated to determine cutoff points. RESULTS: Compared to patients with T2DM, diabetic patients with subclinical atherosclerosis (SA) had a greater CV risk, as measured by the FGCRS, accompanied by markedly decreased microcirculation perfusion. Microcirculatory parameters (TVD and PVD), including carotid intima-media thickness (IMT), brachial-ankle pulse wave velocity (ba-PWV), and FGCRS, were closely associated with SA incidence. Microcirculatory parameters, Index (DMSA screen), and cut-off points were used to screen for SA in patients with T2DM. Furthermore, a new set of four factors identified through machine learning showed optimal sensitivity and specificity for detecting CKM risk in patients with T2DM. Decreased microcirculatory perfusion served as a useful early marker for CKM syndrome risk stratification in patients with T2DM without SA. CONCLUSIONS: Sublingual microcirculatory dysfunction is closely correlated with the risk of SA and CKM risk in T2DM patients. Sublingual microcirculation could be a novel tool for assessing the CKM syndrome stage in patients with T2DM.

Clinical Characteristics of Target Organ Damage in Primary Aldosteronism with or without Metabolic Syndrome.

The aim of this study is to investigate the prevalence of metabolic disorders in patients with primary aldosteronism (PA) and target organ damage (TOD) in different subtypes of patients with PA with or without metabolic syndrome (MS). Patients with PA were screened out from those with secondary hypertension and then subtyped via adrenal venous sampling (AVS). Baseline clinical characteristics (blood pressure, blood glucose, abdominal circumference, and lipid profile) were collected for the diagnosis of MS. Organ damage was evaluated according to cardiac and carotid ultrasound and urine microalbumin measurements. In all 261 patients with PA, 113 patients had concomitant MS and experienced more severe cardiac hypertrophy and increased intima-media thickness (IMT). The incidence of MS and diabetes mellitus (DM) had no statistic difference between the two groups, moreover, the rates of TOD were not different except microalbuminuria. However, metabolic disorders caused more remarkable TOD in PA patients with unilateral hypersecretion. It showed that cardiac hypertrophy was associated with obesity while microalbuminuria was related to plasma aldosterone concentration (PAC) in PA patients. In this retrospective study, our findings suggest that the effect of metabolic disorders on organ damage is more remarkable in patients with unilateral PA.

Activation of Glucagon-Like Peptide-1 Receptor Ameliorates Cognitive Decline in Type 2 Diabetes Mellitus Through a Metabolism-Independent Pathway.

Background Patients with hypertension and diabetes mellitus are susceptible to dementia, but regular therapy fails to reduce the risk of dementia. Glucagon-like peptide-1 receptor agonists have neuroprotective effects in experimental studies. We aimed to assess the effect of liraglutide, a glucagon-like peptide-1 receptor agonist, on cognitive function and whether its effect was associated with metabolic changes in patients with type 2 diabetes mellitus. Methods and Results Fifty patients with type 2 diabetes mellitus were recruited in this prospective study. All patients underwent cognitive assessment and brain activation monitoring by functional near-infrared spectroscopy. At 12 weeks, patients in the glucagon-like peptide-1 group acquired better scores in all cognitive tests and showed remarkable improvement in memory and attention (P=0.040) test compared with the control group after multivariable adjustment. Compared with the control group, liraglutide significantly increased activation of the dorsolateral prefrontal cortex and orbitofrontal cortex brain regions (P=0.0038). After liraglutide treatment, cognitive scores were significantly correlated with changes in these activating brain regions (P<0.05), but no correlation was observed between the changes in cognitive function and changes of body mass index, blood pressure, and glycemic levels. Conclusions We concluded that liraglutide improves cognitive decline in patients with type 2 diabetes mellitus. This beneficial effect is independent of its hypoglycemic effect and weight loss. The optimal intervention should be targeted to cognitive decline in the early stages of dementia. Registration URL: https://www.ClinicalTrials.gov; Unique identifier: NCT03707171.

Adrenal artery ablation for primary aldosteronism without apparent aldosteronoma: An efficacy and safety, proof-of-principle trial.

Primary aldosteronism (PA) is associated with resistant hypertension and cardiovascular events. There are some limitations of current medical and surgical therapies for PA. To determine the efficacy and safety of catheter-based adrenal artery ablation for treatment of PA patients who refused both surgery and medical therapy, we performed this prospective cohort study. Thirty-six PA patients without apparent aldosteronoma were treated by adrenal artery ablation. Primary outcome was postoperative blood pressure and defined daily dose (DDD) of antihypertensive medications after adrenal ablation. Secondary outcome was biochemical success. We assessed outcomes based on Primary Aldosteronism Surgical Outcome (PASO) criteria. Adrenal CT scan, biochemical evaluation, adrenal artery ablation and adrenal venous sampling (AVS) were underwent. After adrenal ablation, complete clinical success (normotension without antihypertensive medication) was achieved in 9/36 (25.0%) patients and partial clinical success (reduction in blood pressure or less antihypertensive medication) in 13/36 (36.1%) patients. Complete biochemical success (correction of hypokalemia and normalization of aldosterone-to-renin ratio) was achieved in 16/36 (44.4%) patients. Office-based and ambulatory blood pressures were reduced by 17/7 and 11/2 mmHg at 6 months after ablation, respectively. The plasma cortisol level in the ablation group decreased slightly, but no patient developed hypoadrenocorticism. Catheter-based adrenal ablation appears to produce substantial and sustained blood pressure reduction and biochemical improvement, with only minor adverse events in PA patients without apparent aldosteronoma. This therapy could be an important supplement for current PA treatments.

Optimization of ASE and SPE conditions for the HPLC-FLD detection of piperazine in chicken tissues and pork.

Accelerated solvent extraction (ASE) and solid-phase extraction (SPE) conditions were optimized by a high-performance liquid chromatography-fluorescence detector (HPLC-FLD) method for the detection of piperazine in chicken tissues and pork. Piperazine residues were determined by precolumn derivatization with trimethylamine and dansyl chloride. Samples were extracted with 2% formic acid in acetonitrile using an ASE apparatus and purified using a Strata-X-C SPE column. The monosubstituted product of the reaction of piperazine with dansyl chloride was 1-dansyl piperazine (1-DNS-piperazine). Chromatographic separations were performed on an Athena C18 column (250 × 4.6 mm, id: 5 µm) with gradient elution using ultrapure water and acetonitrile (5:95, V/V) as the mobile phase. The calibration curves showed good linearity over a concentration range of LOQ-200.0 µg/kg with a coefficient of determination (R2 ) ≥ .9992. The recoveries and relative standard deviations (RSD values) ranged from 78.49% to 97.56% and 1.19% to 5.32%, respectively, across the limit of quantification (LOQ) and 0.5, 1, and 2.0 times the maximum residue limit (MRL; µg/kg). The limits of detection (LODs) and LOQs were 0.96 to 1.85 µg/kg and 3.20 to 5.50 µg/kg, respectively. The method was successfully applied for the validation of animal products in the laboratory.

Quantitative analysis of chloramphenicol, thiamphenicol, florfenicol and florfenicol amine in eggs via liquid chromatography-electrospray ionization tandem mass spectrometry.

A widely applicable method involving liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (LC-ESI/MS/MS) was developed for the simultaneous determination of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in eggs. Samples were extracted with ethyl acetate-acetonitrile-ammonium hydroxide (49:49:2, v/v) and defatted with hexane saturated with acetonitrile. The analysis was carried out on a mass spectrometer via an electrospray interface operated in the positive and negative ionization modes using deuterated chloramphenicol-d5 as the internal standard. The limits of detection and limits of quantification were 0.04-0.5 µg/kg and 0.1-1.5 µg/kg in eggs, respectively. The average recoveries of the four analytes from egg samples were 90.84-108.23%, with relative standard deviations of less than 9.61%. The corresponding intra-day and inter-day variations were found to be less than 8.11% and 11.30%, respectively. Finally, the new approach was successfully applied to the quantitative determination of these analytes in 50 commercial eggs from local supermarkets.

Development of an ASE-GC-MS/MS method for detecting dinitolmide and its metabolite 3-ANOT in eggs.

An accelerated solvent extraction coupled with gas chromatography-tandem mass spectrometry (ASE-GC-MS/MS) method for detecting dinitolmide residue and its metabolite (3-amino-2-methyl-5-nitrobenzamide, 3-ANOT) in eggs was developed and optimized. The samples were extracted using ASE with acetonitrile as the extractant and were purified by passage through a neutral alumina solid-phase extraction column. Then, the samples were analyzed using the GC-MS/MS method. The optimized method parameters were validated according to the requirements set forth by the European Union and the Food and Drug Administration. The average recoveries of dinitolmide and 3-ANOT from eggs (egg white, egg yolk, and whole egg) at the limit of quantification (LOQ), 0.5 maximum residue limit (MRL), 1 MRL, and 2 MRL were 82.74% to 87.49%, the relative standard deviations (RSDs) were less than 4.63%, and the intra-day RSDs and the inter-day RSDs were 2.96% to 5.21% and 3.94% to 6.34%, respectively. The limits of detection and the LOQ were 0.8 to 2.8 µg/kg and 3.0 to 10.0 µg/kg, respectively. The decision limits (CCα ) were 3001.69 to 3006.48 µg/kg, and the detection capabilities (CCß ) were 3001.74 to 3005.22 µg/kg. Finally, the new method was successfully applied to the quantitative determination of dinitolmide and 3-ANOT in 50 commercial eggs from local supermarkets.

[Knockdown of dachshund homolog 1 (DACH1) promotes cell apoptosis and inhibits the invasion and migration abilities of Capan-1 pancreatic cancer cells].

Objective To investigate the impact of the decreased expression of dachshund homolog 1 (DACH1) on cell cycle, apoptosis, invasion and migration of Capan-1 pancreatic cancer cells. Methods After four pairs of DACH1 siRNA were designed and synthesized, double-stranded short hairpin RNA (shRNA) were annealed and inserted into pGenesil-1 vector. The product was then confirmed by enzyme digestion and sequencing analysis. The recombinant plasmids were transfected into Capan-1 cells via Lipofectamine(TM) 2000. Fluorescence microscopy, reverse transcription PCR (RT-PCR) and Western blotting were used to detect the transfection efficiency. Cell apoptosis and cell cycle were tested by flow cytometry. Transwell(TM) assay was used to monitor the invasion and migration abilities of Capan-1 cells. Results Recombinant plasmid pshRNA-DACH1 was successfully constructed and transfected into Capan-1 cells. After transfection, the expression of DACH1 was reduced to some extent. Flow cytometry revealed that cell apoptosis was promoted in the pshRNA-DACH1 transfected group compared with control groups, whereas cell cycle had no significant differences among the groups. Transwell(TM) assay validated that the abilities of migration and invasion were inhibited in the pshRNA-DACH1 transfected group. Conclusion Knockdown of DACH1 expression can remarkably enhance the cell apoptosis, restrain the proliferation, migration and invasion of Capan-1 cells.

Inhibition of DACH1 activity by short hairpin RNA represses cell proliferation and tumor invasion in pancreatic cancer.

Cancer of the pancreas is one of the most lethal diseases worldwide. Better understanding of the molecular mechanisms involved in tumorigenesis is of great consequence to elevate the survival rate. Human Dachshund homologue 1 (DACH1) plays a controversial role in human malignancy progression with its expression being altered in a variety of cancers. Nevertheless, its functional roles and molecular mechanisms in pancreatic cancer remain unknown. The expression of DACH1 in pancreatic cancer cell lines and the ductal epithelial cells were evaluated both at mRNA and protein levels. Three pairs of siRNA targeting the DACH1 gene were designed and synthesized, double-stranded short hairpin RNA (shRNA) were annealed and inserted into pGenesil-1 vector, which was confirmed by enzymatic digestion and sequencing analyses. The successfully constructed recombinant plasmids were transfected into Capan-1 cells and our data indicated that knockdown of DACH1 gene expression showed strong correlation with repressing tumorigenesis. The proliferation of Capan-1 cells was significantly repressed as evaluated by CCK-8 and colony formation assays. Flow cymetry revealed that cell apoptosis was promoted in interference plasmid group compared with control groups (P<0.05), whereas cell cycle had no significant differences among the groups (P>0.05). Transwell assay validated the abilities of migration and invasion as being significantly reduced in pshRNA-DACH1 group. Furthermore, our study suggested that DACH1 expression regulates the pancreatic cancer cell apoptosis through interacting with Bcl-2 signaling axis, whereas it controls cell migration and invasion via epithelial-mesenchymal transition (EMT) process.

Protocadherin-10 acts as a tumor suppressor gene, and is frequently downregulated by promoter methylation in pancreatic cancer cells.

Protocadherin-10 (PCDH10), a member of non-clustered protocadherin family which plays important roles in calcium-dependent cell-cell signal transduction and adhesion. PCDH10 functions as a tumor suppressor gene and its expression is downregulated by promoter methylation in many malignances. In the present study, we explored PCDH10 expression and promoter methylation status, and its biological effects in pancreatic cancer cells, and furthermore, we explored the mechanism of PCDH10 preliminary in pancreatic cancer cells. the mRNA level of PCDH10 was detected by semi-quantitative reverse transcription PCR and promoter methylation status examined by methylation-specific PCR in the pancreatic cancer cells (Capan-1, Panc-1, AsPC-1 and BxPC-3) as well as the human normal pancreatic ductal epithelial cells HPDE6-C7 which was used as a control. The human pancreatic cells were transfected with plasmid pcDNA3.1-PCDH10 or pcDNA3.1 by lipofectamine 2000. The biological function of PCDH10 in pancreatic cancer cells was determined by CCK-8 assay, colony formation assay, flow cytometry, Transwell invasion assay and wound-healing assay. The levels of apoptosis related proteins were examined by western blotting. PCDH10 expression was obviously downregulated in the pancreatic cancer cells (Capan-1, Panc-1, BxPC-3) compared to the normal pancreatic ductal epithelial cells. PCDH10 promoter methylation was observed in the two pancreatic cancer cells Capan-1 and BxPC-3,and the expression of PCDH10 could be restored after treating with 5-aza-2'-deoxycytidine and trichostatin A in the two cell types. Overexpression of PCDH10 can inhibit the proliferation, migration, invasion ability of pancreatic cancer cells and induce apoptosis. Ectopic expression of PCDH10 could increase the levels of PARP, caspase-3, caspase-9 and decrease the level of bcl-2, AKT and p-AKT. PCDH10 acts as a tumor suppressor gene, and is frequently down-regulated by promoter methylation in pancreatic cancer cells. PCDH10 may induce cancer cell apoptosis via the AKT pathway.

[Protocadherin 10 (PCDH10) inhibits the proliferation, invasion and migration ability of BXPC-3 pancreatic cancer cells].

OBJECTIVE: To explore the expression and biological function of protocadherin 10 (PCDH10) in pancreatic cancer cells. METHODS: Reverse transcription PCR (RT-PCR) was used to detect the expression of PCDH10 in CAPAN-1, PANC-1, ASPC-1, BXPC-3 pancreatic cancer cells and the HPDE6-C7 normal human pancreatic ductal epithelial cells. Recombinant plasmid pcDNA3.1-PCDH10 and empty vector pcDNA3.1 were transfected into BXPC-3 pancreatic cancer cells via Lipofectamine(TM)2000. After transfection, the mRNA expression of PCDH10 was examined by RT-PCR, and the protein level was detected by Western blotting. CCK-8 and colony formation assays were used to analyze the cell proliferation. Cell apoptosis was tested by flow cytometry combined with annexin V-FITC/PI staining. Transwell(TM) and wound healing assays were performed to measure the invasion and migration ability of the cells. RESULTS: Compared with the normal pancreatic ductal epithelial cells, the expression of PCDH10 was obviously lower in the CAPAN-1, PANC-1, BXPC-3 cells. RT-PCR and Western blotting revealed that PCDH10 expression significantly increased in BXPC-3 cells transfected with plasmid pcDNA3.1-PCDH10 compared with the ones with empty vector pcDNA3.1. CCK-8 and colony formation assays showed that the PCDH10-transfected cells grew more slowly than the empty vector-transfected cells. Annexin V-FITC/PI staining combined with flow cytometry proved that the apoptosis in the PCDH10-transfected cells remarkably increased compared with that in the control group. A reduction of the invasion and migration ability was found obviously in the PCDH10-transfected cells by Transwell(TM) assay. The wound healing assay also showed that the PCDH10-transfected cells spread the more slowly than the empty vector-transfected cells. CONCLUSION: The expression of PCDH10 was down-regulated in the pancreatic cancer cells. PCDH10 over-expression could significantly induce cell apoptosis, and restrain proliferation, invasion and migration ability of BXPC-3 pancreatic cancer cells.