Serum secreted phosphoprotein 1 level is associated with plaque vulnerability in patients with coronary artery disease.

Background: Vulnerable plaque was associated with recurrent cardiovascular events. This study was designed to explore predictive biomarkers of vulnerable plaque in patients with coronary artery disease. Methods: To reveal the phenotype-associated cell type in the development of vulnerable plaque and to identify hub gene for pathological process, we combined single-cell RNA and bulk RNA sequencing datasets of human atherosclerotic plaques using Single-Cell Identification of Subpopulations with Bulk Sample Phenotype Correlation (Scissor) and Weighted gene co-expression network analysis (WGCNA). We also validated our results in an independent cohort of patients by using intravascular ultrasound during coronary angiography. Results: Macrophages were found to be strongly correlated with plaque vulnerability while vascular smooth muscle cell (VSMC), fibrochondrocyte (FC) and intermediate cell state (ICS) clusters were negatively associated with unstable plaque. Weighted gene co-expression network analysis showed that Secreted Phosphoprotein 1 (SPP1) in the turquoise module was highly correlated with both the gene module and the clinical traits. In a total of 593 patients, serum levels of SPP1 were significantly higher in patients with vulnerable plaques than those with stable plaque (113.21 [73.65 - 147.70] ng/ml versus 71.08 [20.64 - 135.68] ng/ml; P < 0.001). Adjusted multivariate regression analysis revealed that serum SPP1 was an independent determinant of the presence of vulnerable plaque. Receiver operating characteristic curve analysis indicated that the area under the curve was 0.737 (95% CI 0.697 - 0.773; P < 0.001) for adding serum SPP1 in predicting of vulnerable plaques. Conclusion: Elevated serum SPP1 levels confer an increased risk for plaque vulnerability in patients with coronary artery disease.

Efficacy of Bleomycin-Lauromacrogol Foam in Pediatric Macrocystic Lymphatic Malformations With and Without Intracapsular Hemorrhage.

BACKGROUND: Sclerotherapy is purportedly less effective in patients with hemorrhagic than with non-hemorrhagic lymphatic malformations (LMs). We aimed to compare the efficacy of bleomycin-lauromacrogol foam (BLF) sclerotherapy in the treatment of macrocystic LMs with and without intralesional hemorrhage. METHODS: Fifty-five children with macrocystic LMs admitted to the Pediatric Surgery Department were retrospectively included. The patients were allocated into a hemorrhage group (23 cases) or a non-hemorrhage group (32 cases) based on the occurrence of an intracapsular hemorrhage. The diagnosis was confirmed by physical examination, color ultrasound, magnetic resonance imaging, and puncture findings. BLF was injected into the capsule after draining the cystic fluid under color ultrasound guidance. Patients whose lesions were unchanged or showed minor change after 1 month were treated again using the same method. Changes in lesion size and the number of treatments were recorded. Effectiveness was classified as excellent (volume reduction ≥90%), good (50%≤volume reduction<90%), or poor (volume reduction <50%). RESULTS: In the hemorrhage group, 17, 6, and 0 patients' outcomes were classified as excellent, good, and poor, respectively. The overall efficacy rate was 100%. In the non-hemorrhage group, 23, 7, and 2 patients' outcomes were classified as excellent, good, and poor, respectively. The overall efficacy rate was 93.8%. There was no significant difference in efficacy rate between groups (P = 0.767). CONCLUSIONS: BLF is an effective and safe treatment for macrocystic LMs with bleeding. The results were similar in patients with and without bleeding. LEVEL OF EVIDENCE: Treatment, Level III.

NLRP3 activation in macrophages promotes acute intestinal injury in neonatal necrotizing enterocolitis.

BACKGROUND: Macrophages are involved in various immune inflammatory disease conditions. This study aimed to investigate the role and mechanism of macrophages in regulating acute intestinal injury in neonatal necrotizing enterocolitis (NEC). METHODS: CD68, nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing 3 (NLRP3), cysteine aspartate-specific protease-1 (caspase-1), and interleukin-1ß (IL-1ß) in paraffin sections of intestinal tissues from NEC and control patients were detected with immunohistochemistry, immunofluorescence, and western blot. Hypertonic pet milk, hypoxia and cold stimulation were used to establish a mouse (wild type and Nlrp3-/-) model of NEC. The mouse macrophage (RAW 264.7) and rat intestinal epithelial cell-6 lines were also cultured followed by various treatments. Macrophages, intestinal epithelial cell injuries, and IL-1ß release were determined. RESULTS: Compared to the gut "healthy" patients, the intestinal lamina propria of NEC patients had high macrophage infiltration and high NLRP3, caspase-1, and IL-1ß levels. Furthermore, in vivo, the survival rate of Nlrp3-/- NEC mice was dramatically improved, the proportion of intestinal macrophages was reduced, and intestinal injury was decreased compared to those of wild-type NEC mice. NLRP3, caspase-1, and IL-1ß derived from macrophages or supernatant from cocultures of macrophages and intestinal epithelial cells also caused intestinal epithelial cell injuries. CONCLUSIONS: Macrophage activation may be essential for NEC development. NLRP3/caspase-1/IL-1ß cellular signals derived from macrophages may be the underlying mechanism of NEC development, and all these may be therapeutic targets for developing treatments for NEC.

Laparoscopic approach for managing intussusception in children: Analysis of 65 cases.

BACKGROUND: Intussusception can be managed by pneumatic reduction, ultrasound-guided hydrostatic reduction, open or laparoscopic surgery, but laparoscopy in such cases remains controversial. AIM: To explore the clinical characteristics, effectiveness, and complications of surgical reduction for intussusception using laparoscopy in children. METHODS: This study was a retrospective case series of pediatric patients with intussusception who underwent surgical reduction by laparoscopy from May 2011 to April 2016 at Taizhou Hospital of Zhejiang Province. Clinical characteristics (operation time, intraoperative blood loss, conversion rate of laparotomy, reasons for conversion, postoperative hospital stay, and adverse events) were described. RESULTS: The 65 patients included 45 boys and 20 girls. The average age was 2.3 years (27.5 ± 24.5 mo). Of the 65 patients, 61 underwent surgical reduction by laparoscopy after a failed enema reduction of intussusception, and four underwent the procedure directly. All patients were treated successfully and 57 (87.7%) patients underwent successful laparoscopic surgery, two of which had a spontaneous reduction. Among the remaining cases, one was converted to open surgery via right upper quadrant incision, and seven required enlarged umbilical incisions. Intestinal resection was performed in 5 patients because of abnormal bowel lesions. There were no complications (intestinal perforations, wound infections, or intestinal adhesions) during the follow-up of 3 years to 8 years. Two patients experienced a recurrence of intussusception; one was resolved with pneumatic reduction, and the other underwent a second laparoscopic surgery. CONCLUSION: Laparoscopic approach for pediatric intussusception is feasible and safe. Bowel resection if required can be performed by extending umbilical incision without the conventional laparotomy.

Single-Site Umbilical Laparoscopic Pyloromyotomy Using a Pyloric Electrocoagulation Chisel Combined with a Left-Handed Main Operation for Congenital Hypertrophic Pyloric Stenosis.

Background: Laparoscopic pyloromyotomy has become a gold standard for the treatment of congenital hypertrophic pyloric stenosis (HPS). There have been recent reports on the use of transumbilical single-site laparoscopic surgery for congenital HPS; however, using transumbilical single-site laparoscopic surgery in pediatric cases is still controversial due to the difficulty with manipulation. In this study, some preliminary experience with the application of a novel transumbilical single-site laparoscopic approach in congenital HPS is described. Methods: A retrospective study was conducted involving 25 patients with congenital HPS treated in our hospital from August 2016 to August 2019. A pyloric electrocoagulation chisel combined with a left-handed main operation was completed in all of the patients and the operative times, postoperative length of stay, and operative complications were recorded. Results: The laparoscopic operation was completed in 25 patients with an average operative time of 21.9 ± 5.5 minutes, average postoperative length of stay of 2.5 ± 0.9 days, and no perforations of the pyloric mucosa, recurrent obstruction, surgical incision infections, and incision hernias. All of the patients had at least 3 months of follow-up, good growth and development, and the parents were satisfied with the postoperative scars. Conclusion: A pyloric electrocoagulation chisel combined with a left-handed main operation in the treatment of congenital HPS by a single-site umbilical laparoscopic pyloromyotomy is safe and effective, and can achieve a satisfactory cosmetic effect.

Secretory vimentin is associated with coronary artery disease in patients and induces atherogenesis in ApoE-/- mice.

PURPOSE: The present study aimed to investigate the relationship between serum levels of secretory vimentin and coronary artery disease (CAD). The biological effect of secretory vimentin was ascertained by experiments. METHODS: We analysed serum levels of secretory vimentin in CAD patients (n = 288) and non-CAD controls (n = 195) by ELISA. To evaluate the pro-inflammatory effects of secreted vimentin, the human aortic endothelial cells (HAECs) and human peripheral blood mononuclear cells (PBMCs) were treated with recombinant vimentin or saline. Intraperitoneal injection of vimentin (1 µg/each) or saline was performed every other day for 12 weeks in ApoE-/- mice for assessment of atherogenic effect. RESULTS: Serum levels of secretory vimentin were significantly increased in CAD patients than in health controls (p < 0.05), and correlated with the number of diseased coronary arteries, Syntax and Gensini score (for all comparison, p < 0.01). Logistic regression analysis showed that vimentin level is an independent determinant of CAD. In experiments, recombinant vimentin protein enhanced the expression of adhesion molecules and inflammatory cytokines in both endothelial cells and macrophages. This protein also promoted macrophage-endothelial cells adhesion in vitro and the recruitment of leukocytes to mesenteric venules in C57BL/6 mice. Compared with saline, intraperitoneal injection of recombinant vimentin (1 µg/each) every other day induced atherogenesis in ApoE-/- mice at 12-weeks, with significant increase of inflammatory cytokine and adhesion molecules expression in aortic tissue (p < 0.05). CONCLUSION: Serum vimentin levels are associated with the presence and the severity of CAD. Vimentin protein promotes atherogenesis in ApoE-/- mice.

The Application of Pyloric chisel in the Treatment of Hypertrophic Pyloric Stenosis by Single-Site Umbilical Laparoscopic Pyloromyotomy.

OBJECTIVE: The feasibility and perspective of pyloric chisel were discussed through the comparison of pyloric chisel and knife in the treatment of hypertrophic pyloric stenosis (HPS) in single-site umbilical laparoscopic pyloromyotomy (SSULP). METHODS: Fifty-eight cases of HPS treated in our hospital from February 2011 to March 2016 were retrospectively analyzed, in which 30 patients underwent pyloric chisel (Pyloric chisel Group) and 28 patients underwent knife (Knife Group). Operative time, estimated blood loss, and complications between the two groups were analyzed. RESULTS: The operative time was shorter in Pyloric chisel Group than Knife Group (P < .05). The estimated blood loss was lower in Pyloric chisel Group than Knife Group (P < .05). The complication was less in Pyloric chisel Group than Knife Group (P < .05). There were 2 cases of mucosal perforations requiring conversions to open in Knife Group. Five cases of serous tearing occurred in the Knife Group. There was 1 case of serous tearing in the Pyloric chisel Group. All patients were followed up for 3 months, and there was no distinct scar in the umbilical. CONCLUSIONS: Patients were satisfied with no distinct scars in abdominal wall by pyloric chisel or knife to treat HPS in SSULP, but pyloric chisel is more effective and safer.

[Angiotensin II induced cardiac hypertrophy is blocked by PTEN via suppressing Ca2+/Calcineurin pathway].

OBJECTIVE: To investigate the effects of PTEN on Ang II induced cardiomyocyte hypertrophy and subsequent Ca(2+)/Calcineurin pathway changes. METHODS: Primary cultured neonatal rat cardiomyocytes were cultured and were treated with phosphate-buffered saline, empty adenovirus (Ad-GFP), or adenovirus encoding for PTEN (Ad-PTEN-GFP) for 48 h and Ang II (10(-7) mol/L) was added to the medium for another 24 h. Cells were harvested and intracellular Ca(2+) concentration ([Ca(2+)] i) was determined by Fura-2/AM ratio imaging analysis; PTEN, ANF, beta-MHC and CaNAbeta mRNA evaluated with RT-PCR; PTEN and CaNAbeta protein by Western blot; CaN phosphatase activity by CaN detecting kits. RESULTS: PTEN at mRNA and protein levels were significantly higher in Ad-PTEN-GFP treated cardiomyocytes than that of Ad-GFP treated cardiomyocytes. Ang II stimulation upregulated [Ca(2+)] i, CaNAbeta at mRNA and protein levels and CaN phosphatase activity in Ad-GFP treated cardiomyocytes but not in Ad-PTEN-GFP treated cardiomyocytes. CONCLUSIONS: Cardiac hypertrophy induced by Ang II could be blocked by PTEN overexpression via suppressing Ca(2+)/Calcineurin pathway.

[PTEN negatively regulates isoproterenol-induced cardiac hypertrophy and effects of captopril on PTEN expression].

OBJECTIVE: To examine the negative regulation role of PTEN in isoproterenol-induced cardiac hypertrophy by testing the expression of PTEN mRNA and protein and to explore the effects of captopril (Cap) on PTEN expression. METHODS: Twenty four rats were randomly divided into three groups: control group, ISO group, and ISO+Cap group. The following parameters were examined:body weight (BW), heart weight (HW), left ventricular weight (LVW), left ventricular end-diastolic pressure (LVEDP), left ventricular end-systolic pressure (LVESP) and +/- dp/dt(max). The ratio of HW/BW and LVW/BW was calculated. PTEN mRNA and protein were tested by RT-PCR and Western blot, respectively. RESULTS: (1) Compared with the control group, the ratio of HW/BW and LVW/BW, LVEDP and LVESP were all increased in ISO group and ISO+Cap group (P < 0.05), but +/- dp/dt(max) was decreased (P < 0.05); (2) compared with the ISO group, the ratio of HW/BW and LVW/BW, LVEDP, LVESP were all decreased in ISO+Cap group (P < 0.05), but +/- dp/dt(max) was increased (P < 0.05); (3) compared with the control group, PTEN mRNA and protein were up-regulated in ISO group and ISO+Cap group; (4) compared with the ISO group, PTEN mRNA and protein were up-regulated in ISO+Cap group. CONCLUSIONS: PTEN mRNA and protein are up-regulated in isoproterenol-induced cardiac hypertrophy. Captopril can up-regulate PTEN expression in cardiac hypertrophy. There is a negative regulative mechanism in cardiac hypertrophy process, in which PTEN is probably an endogenous negative regulator of cardiac hypertrophy.

[The relationship between expressions of beta1-, beta2-, beta3-adrenoceptor mRNA of myocardium and cardiac function in patients with heart failure].

OBJECTIVE: To investigate the alteration of expressions of beta(1)-, beta(2)-, beta(3)-adrenoceptor mRNA in human myocardial tissue and the relation between their expressions and cardiac function in patient with heart failure. METHODS: The mRNA expressions of beta(1)-, beta(2)- and beta(3)-adrenergic receptors in myocardial tissue were analyzed by using the reverse transcriptase-polymerase chain reaction in 24 patients with heart failure of valvular heart disease and 5 control subjects. RESULTS: Beta(1)-adrenergic receptor mRNA expressions in myocardium were significantly lower in patients with heart failure than those in control subjects, and progressively reduced with aggravation of heart function. By contrast, beta(3)-adrenoceptor mRNA expressions were significantly higher in patients with heart failure than those in controls, and progressively elevated with aggravation of cardiac function. No difference was observed in beta(2)-adrenergic receptor among all groups. CONCLUSION: The changes of beta-adrenergic receptor mRNA expression are associated with the severity of heart failure.

[CaN-NFAT3 signal pathway: a crucial hinge relates Ca2+ signal with cardiomyocyte hypertrophy].

OBJECTIVE: To investigate the role of [Ca2+]i from different origins in the course of myocardial hypertrophy mediated by CaN-NFAT3 signal transduction. METHODS: The primarily cultured cardiomyocyte were irritated with angiotensin (Ang) II and ryanodine (RY) which cause Ca2+ inflow and release respectively. Then to observe the changes of CaN-NFAT3 pathway were then observed. Western blotting was employed to semi-quantify CaN, NFAT3 and GATA4. The distribution of NFAT3 was shown with immunocytochemistry, (3)H-Leu incorporation was used as an index of myocyte hypertrophy. Cyclosporin A (CsA) was applied to restrain CaN-NFAT3 pathway as a kind of CaN-selective antagonist. RESULTS: CaN, NFAT3, GATA4 expression significantly increased 1 and 3 days after the stimulation of cardiomyocytes with Ang II and RY (10(-7) mol/L) as compared with that of a control group (P > 0.05) and (3)H-Leu incorporation distinctly increased after Ang II and RY (10(-7) mol/L) stimulation (P > 0.05 versus control group). On the first day of Ang II and RY stimulation, NFAT3 was shown mainly as intra-nuclear expression rather than cytoplasmic expression as seen in the control group. All of the above effects were suppressed by CsA administration, but they were rarely suppressed if CsA was not administered (P > 0.05). CONCLUSIONS: It is shown that both Ca2+ inflow and release may activate CaN-NFAT3 signal pathway, which responds to increase of [Ca2+]i and is independent of its origin, indicating the augment of [Ca2+]i may trigger CaN-NFAT3 signal transduction and consequently induce myocyte hypertrophy. Moreover, CsA may restrain the expression and activation of CaN-NFAT3 and protein synthesis of myocytes in response to Ang II and RY stimulation.