Plasminogen Activator inhibitor-I Research Articles

Antenatal maternal serum biomarkers as a predictor for placenta accreta spectrum disorders

IntroductionPlacenta accreta spectrum (PAS) disorder, an abnormal adherence of the placenta to the uterine wall, with variable degrees of invasion, is a major cause of maternal morbidity and mortality associated with severe postpartum hemorrhage. PAS is diagnosed using ultrasonography or with magnetic resonance imaging; in many centers there is a lack of PAS diagnostic expertise in diagnosing. Hence, we investigated the performance of selected maternal plasma protein biomarkers, antithrombin-III (AT-3), plasminogen activator inhibitor-I (PAI-I), soluble vascular endothelial growth factor receptor-II (sVEGFR-2), and soluble Tie-II (sT-2) for prenatal screening in pregnancies at a high risk of PAS. MethodsThis prospective study, conducted in a tertiary hospital from September 2021 to May 2022, included pregnant women with placenta previa suspicious of PAS between 28 and 42 weeks of gestation. Four serum samples were collected from each woman to evaluate serum concentrations and compared between placenta previa (control) and PAS groups. The screening performances of the biomarkers were analyzed, and the best screening model for PAS was created. ResultsTwenty-two women with PAS and 18 with placenta previa alone were included (n = 40). The median concentrations of PAI-I, AT-3, sVEGFR-2, and sT-2 among the PAS group were 21.2, 6154.6, 7.5, and 12.8 ng/mL, respectively. The best screening model for PAS combined all four biomarkers with a history of cesarean delivery (77 % sensitivity, 89 % specificity, and an AUC of 0.87). DiscussionA combination of the four maternal serum biomarkers in women with a history of cesarean delivery presented the most promising model for prenatal screening of PAS. ConclusionA combination of the four maternal serum biomarkers with a history of cesarean delivery presented the most promising model for prenatal screening of PAS.

Impact of Standard Versus Low Pneumoperitoneum Pressure on Peritoneal Environment in Laparoscopic Cholecystectomy. Randomized Clinical Trial.

High CO 2 pneumoperitoneum pressure during laparoscopy adversely affects the peritoneal environment. This study hypothesized that low pneumoperitoneum pressure may be linked to less peritoneal damage and possibly to better clinical outcomes. One hundred patients undergoing scheduled laparoscopic cholecystectomy were randomized 1:1 to low or to standard pneumoperitoneum pressure. Peritoneal biopsies were performed at baseline time and 1 hour after peritoneum insufflation in all patients. The primary outcome was peritoneal remodeling biomarkers and apoptotic index. Secondary outcomes included biomarker differences at the studied times and some clinical variables such as length of hospital stay, and quality and safety issues related to the procedure. Peritoneal IL6 after 1 hour of surgery was significantly higher in the standard than in the low-pressure group (4.26±1.34 vs. 3.24±1.21; P =0.001). On the contrary, levels of connective tissue growth factor and plasminogen activator inhibitor-I were higher in the low-pressure group (0.89±0.61 vs. 0.61±0.84; P =0.025, and 0.74±0.89 vs. 0.24±1.15; P =0.028, respectively). Regarding apoptotic index, similar levels were found in both groups and were 44.0±10.9 and 42.5±17.8 in low and standard pressure groups, respectively. None of the secondary outcomes showed differences between the 2 groups. Peritoneal inflammation after laparoscopic cholecystectomy is higher when surgery is performed under standard pressure. Adhesion formation seems to be less in this group. The majority of patients undergoing surgery under low pressure were operated under optimal workspace conditions, regardless of the surgeon's expertise.

Diagnostic Power of Circulatory Metabolic Biomarkers as Metabolic Syndrome Risk Predictors in Community-Dwelling Older Adults in Northwest of England (A Feasibility Study).

Background: Metabolic Syndrome (MetS) is a cluster of risk factors for diabetes and cardiovascular diseases with pathophysiology strongly linked to aging. A range of circulatory metabolic biomarkers such as inflammatory adipokines have been associated with MetS; however, the diagnostic power of these markers as MetS risk correlates in elderly has yet to be elucidated. This cross-sectional study investigated the diagnostic power of circulatory metabolic biomarkers as MetS risk correlates in older adults. Methods: Hundred community dwelling older adults (mean age: 68.7 years) were recruited in a study, where their blood pressure, body composition and Pulse Wave Velocity (PWV) were measured; and their fasting capillary and venous blood were collected. The components of the MetS; and the serum concentrations of Interleukin-6 (IL-6), Tumor Necrosis Factor-α (TNF-α), Plasminogen Activator Inhibitor-I (PAI-I), Leptin, Adiponectin, Resistin, Cystatin-C, C-Reactive Protein (CRP), insulin and ferritin were measured within the laboratory, and the HOMA1-IR and Atherogenic Index of Plasma (AIP) were calculated. Results: Apart from other markers which were related with some cardiometabolic (CM) risk, after Bonferroni correction insulin had significant association with all components of Mets and AIP. These associations also remained significant in multivariate regression. The multivariate odds ratio (OR with 95% confidence interval (CI)) showed a statistically significant association between IL-6 (OR: 1.32 (1.06–1.64)), TNF-α (OR: 1.37 (1.02–1.84)), Resistin (OR: 1.27 (1.04–1.54)) and CRP (OR: 1.29 (1.09–1.54)) with MetS risk; however, these associations were not found when the model was adjusted for age, dietary intake and adiposity. In unadjusted models, insulin was consistently statistically associated with at least two CM risk factors (OR: 1.33 (1.16–1.53)) and MetS risk (OR: 1.24 (1.12–1.37)) and in adjusted models it was found to be associated with at least two CM risk factors and MetS risk (OR: 1.87 (1.24–2.83) and OR: 1.25 (1.09–1.43)) respectively. Area under curve (AUC) for receiver operating characteristics (ROC) demonstrated a good discriminatory diagnostics power of insulin with AUC: 0.775 (0.683–0.866) and 0.785 by cross validation and bootstrapping samples for at least two CM risk factors and AUC: 0.773 (0.653–0.893) and 0.783 by cross validation and bootstrapping samples for MetS risk. This was superior to all other AUC reported from the ROC analysis of other biomarkers. Area under precision-recall curve for insulin was also superior to all other markers (0.839 and 0.586 for at least two CM risk factors and MetS, respectively). Conclusion: Fasting serum insulin concentration was statistically linked with MetS and its risk, and this link is stronger than all other biomarkers. Our ROC analysis confirmed the discriminatory diagnostic power of insulin as CM and MetS risk correlate in older adults.

Hyperbaric oxygen therapy affects insulin sensitivity/resistance by increasing adiponectin, resistin, and plasminogen activator inhibitor-I in rats

Background/aim Hyperbaric oxygen therapy (HBOT) causes insulin sensitivity, but the reason for this is not known yet. The aim of the present study was to investigate the effect of HBOT on insulin sensitivity via resistin, plasminogen activator inhibitor-I (PAI - I), and adiponectin.Materials and methods The study was designed using HBOT and control groups, with eight rats in each group. After 20 days of HBOT under 2.5 atmospheres for 90 min, the fasting insulin (FI), resistin, PAI-I, homeostatic model assessment of insulin resistance scores (HOMA–IR), quantitative insulin sensitivity check index (QUICKI), fasting plasma glucose (FPG), triglyceride, and high-density lipoprotein cholesterol (HDL-C) in the plasma were measured. The resistin, PAI-I, and adiponectin mRNA expression levels were also measured in the adipose tissue.ResultsCompared to the control group, the FI, FPG, and HOMA-IR scores were significantly lower in the HBOT group, whereas the HDL-C and QUICKI scores were found to be higher. In addition, the resistin, adiponectin, and PAI-I mRNA expression levels were also higher in the HBOT group.ConclusionThe present study demonstrated that the HBOT had regulated the FI, FPG, and HDL-C associated with metabolic syndrome and diabetes mellitus. Moreover, the study showed that HBOT causes insulin sensitivity by raising adiponectin.

Chronic Low Grade Inflammation in Pathogenesis of PCOS.

Polycystic ovary syndrome (PCOS) is a one of the most common endocrine disorders, with a prevalence rate of 5–10% in reproductive aged women. It’s characterized by (1) chronic anovulation, (2) biochemical and/or clinical hyperandrogenism, and (3) polycystic ovarian morphology. PCOS has significant clinical implications and can lead to health problems related to the accumulation of adipose tissue, such as obesity, insulin resistance, metabolic syndrome, and type 2 diabetes. There is also evidence that PCOS patients are at higher risk of cardiovascular diseases, atherosclerosis, and high blood pressure. Several studies have reported the association between polycystic ovary syndrome (PCOS) and low-grade chronic inflammation. According to known data, inflammatory markers or their gene markers are higher in PCOS patients. Correlations have been found between increased levels of C-reactive protein (CRP), interleukin 18 (IL-18), tumor necrosis factor (TNF-α), interleukin 6 (IL-6), white blood cell count (WBC), monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1α (MIP-1α) in the PCOS women compared with age- and BMI-matched controls. Women with PCOS present also elevated levels of AGEs and increased RAGE (receptor for advanced glycation end products) expression. This chronic inflammatory state is aggravating by obesity and hyperinsulinemia. There are studies describing mutual impact of hyperinsulinemia and obesity, hyperandrogenism, and inflammatory state. Endothelial cell dysfunction may be also triggered by inflammatory cytokines. Many factors involved in oxidative stress, inflammation, and thrombosis were proposed as cardiovascular risk markers showing the endothelial cell damage in PCOS. Those markers include asymmetric dimethylarginine (ADMA), C-reactive protein (CRP), homocysteine, plasminogen activator inhibitor-I (PAI-I), PAI-I activity, vascular endothelial growth factor (VEGF) etc. It was also proposed that the uterine hyperinflammatory state in polycystic ovary syndrome may be responsible for significant pregnancy complications ranging from miscarriage to placental insufficiency. In this review, we discuss the most importance evidence concerning the role of the process of chronic inflammation in pathogenesis of PCOS.

Nano-Curcumin Regulates p53 Phosphorylation and PAI-1 Expression during Bleomycin Induced Injury in Alveolar Basal Epithelial Cells

Background: Bleomycin (BLM) is known to cause DNA damage in the Alveolar Epithelial Cells (AECs). It is reported that BLM is involved in the up-regulation of inflammatory molecules such as neutrophils, macrophages, chemokines and cytokines. The complex underlying mechanism for inflammation mediated progression of lung injury is still unclear. This investigation was designed to understand the molecular mechanisms associated with p53 mediated modulation of Plasminogen Activator Inhibitor-I (PAI-I) expression and its regulation by nano-curcumin formulation. Methods: A549 cells were treated with BLM to cause the cellular damage in vitro and commercially available nano-curcumin formulation was used as an intervention. Cytotoxic effect of nano-curcumin was analyzed using Methyl Thiazolyl Tetrazolium (MTT) assay. Protein expressions were analyzed using western blot to evaluate the p53 mediated changes in PAI-I expression. Results: Nano-curcumin showed cytotoxicity up to 88.5 % at a concentration of 20 μg/ml after 48 h of treatment. BLM exposure to the cells activated the phosphorylation of p53, which in turn increased PAII expression. Nano-curcumin treatment showed a protective role against phosphorylation of p53 and PAI-I expression, which in turn regulated the fibro-proliferative phase of injury induced by bleomycin. Conclusion: Nano-curcumin could be used as an effective intervention to regulate the severity of lung injury, apoptosis of AECs and fibro-proliferation during pulmonary injury.

Markers of blood coagulation and fibrinolysis in patients with early and delayed microsurgical reconstructions in the lower extremities

Background: In this study, markers of coagulation and fibrinolysis were assessed during early and delayed microsurgical reconstruction in patients with traumatic defects of their lower legs to analyse whether an imbalance of the hemostasis after trauma might predispose the development of vascular complications.Methods: The prospective study included 70 patients. In 35 patients, surgery was performed within 72 hours after injury. In 35 other patients, delayed free flap transfer was performed between 14–21 days after trauma. In each group, reconstruction was performed with a fasciocutaneous anterior-lateral thigh flap (ALT, n = 18) or a myocutaneous flap (latissimus dorsi flap; n = 17). Blood samples were collected preoperatively, intraoperatively, and 3, 6, 12, 24, 36, 48, 72, 96 and 120 hours after the operation. Analysed parameters included markers of coagulation such as prothrombin fragment 1 + 2 (F1 + 2), thrombin-antithrombin III-complex (TAT), and antithrombin, as well as fibrinolysis markers such as plasminogenactivator inhibitor-I (PAI-1), tissue-plasminogenactivator (t-PA), and plasminogen.Results: Preoperatively, levels of F1 + 2, TAT, and PAI-1 were significantly higher in patients with delayed reconstruction (p < .05). Patients with later vascular complications in this group (n = 5) presented a significant higher concentration of TAT, F1 + 2, and PAI-1 (p < .05). Twelve and 24 hours after free flap surgery, patients with vascular complications presented significant elevated levels of these markers (p < .05).Conclusions: Patients with delayed free flap surgery after lower leg trauma present a hypercoagulable state in their blood due to activation of the coagulation system and hypofibrinolysis. Early reconstruction might minimise the risk of flap failure caused by hypercoagulability.

Role of ID Proteins in BMP4 Inhibition of Profibrotic Effects of TGF-β2 in Human TM Cells

PurposeIncreased expression of TGF-β2 in primary open-angle glaucoma (POAG) aqueous humor (AH) and trabecular meshwork (TM) causes deposition of extracellular matrix (ECM) in the TM and elevated IOP. Bone morphogenetic proteins (BMPs) regulate TGF-β2–induced ECM production. The underlying mechanism for BMP4 inhibition of TGF-β2–induced fibrosis remains undetermined. Bone morphogenic protein 4 induces inhibitor of DNA binding proteins (ID1, ID3), which suppress transcription factor activities to regulate gene expression. Our study will determine whether ID1and ID3 proteins are downstream targets of BMP4, which attenuates TGF-β2 induction of ECM proteins in TM cells.MethodsPrimary human TM cells were treated with BMP4, and ID1 and ID3 mRNA, and protein expression was determined by quantitative PCR (Q-PCR) and Western immunoblotting. Intracellular ID1 and ID3 protein localization was studied by immunocytochemistry. Transformed human TM cells (GTM3 cells) were transfected with ID1 or ID3 expression vectors to determine their potential inhibitory effects on TGF-β2–induced fibronectin and plasminogen activator inhibitor-I (PAI-1) protein expression.ResultsBasal expression of ID1-3 was detected in primary human TM cells. Bone morphogenic protein 4 significantly induced early expression of ID1 and ID3 mRNA (P < 0.05) and protein in primary TM cells, and a BMP receptor inhibitor blocked this induction. Overexpression of ID1 and ID3 significantly inhibited TGF-β2–induced expression of fibronectin and PAI-1 in TM cells (P < 0.01).ConclusionsBone morphogenic protein 4 induced ID1 and ID3 expression suppresses TGF-β2 profibrotic activity in human TM cells. In the future, targeting specific regulators may control the TGF-β2 profibrotic effects on the TM, leading to disease modifying IOP lowering therapies.

A nutritional nonalcoholic steatohepatitis minipig model

Background and AimsThe objective of this study was to elucidate whether a Western diet was associated with nonalcoholic steatohepatitis (NASH), and the relationship between NASH, autophagy and endoplasmic reticulum (ER) stress. MethodsFour-month-old Lee–Sung minipigs were randomly assigned to two groups: control diet (C) and Western diet (W), for a 5-month experimental period. ResultsFeeding a Western diet produced a body composition with more fat, less lean and a greater liver weight. Compared with C pigs, W pigs also exhibited an elevated level of plasma insulin and free fatty acid. The W pigs displayed glucose intolerance, lower circulation antioxidant capacity and greater hepatic oxidative stress. Furthermore, pig fed the W diets had increased collagen accumulation in the liver and elevated systemic inflammation [tumor necrosis factor α and interleukin (IL)-6]. Compared with C pigs, W pigs had higher hepatic ER stress-related protein expression of GRP94, CHOP and caspase-12. The W pigs also had greater hepatic autophagy-related protein expression of p62 and LC3II. In an obesity antibody array analysis, W pigs had higher type 2 diabetes mellitus- (insulin-like growth factor 1, osteoprotegerin and resistin), atherosclerosis- (vascular endothelial growth factor, platelet-derived growth factor-AA and plasminogen activator inhibitor-I) and inflammation- [IL-1, macrophage-stimulating protein alpha, X-linked ectodermal dysplasia receptor and serum amyloid A (SAA)] related protein expressions. In addition, W pigs had greater plasma SAA concentration than C pigs and plasma SAA level was highly associated with IL-6. ConclusionsWe successfully established a NASH pig model, and our findings suggested an association of NASH with ER stress and autophagy. The SAA has potential as a novel plasma biomarker for nonalcoholic fatty liver disease pigs.

Specifics of allelic polymorphism of genes associated with increased therombogenesis in pregnant women with fetal loss syndrome

The Up-to-date research of perinatal mortality in Russia has shown change in its structure with increase of proportion of antenatal mortality in the first place, which may be declarative of disorders in fetal vital activities. Reproductive losses in different periods of pregnancy are increasingly frequently united under the term of fetal loss syndrome (FLS), regardless of ethiopathogenic factors. The mostly discussed issue is thrombotic disorders related to inherited and aquired thrombophilia. Research objective was to evaluate the role of adverse allelic variant genes carriage, associated with increased thrombogenesis, in development of FLS. 105 pregnant women with FLS or FLS risk factors in the past have been examined. The women have been affiliated with SPb GBUZ «Women's clininc N 22». The women have been tested for gene mutations in Factor V (FV Leiden), prothrombin, β-subunit of factor I, plasminogen activator inhibitor-I, methylenete-trahydrofolate reductase and glucoprotein IIIa. The study results have shown that 100 per cents of women in the study group with FLS or FLS risk factors in the past have been carriers of at least one allelic variant associated with increased risk of thrombogenesis. A combination of two or more adverse geetic variants (potential risk factors for FLS) have been registered in the overwhelming majority of cases. The incidence of Leiden and genotype 1565CC gene GpIIIa mutations in both study groups was significantly different as compared to the population. The study results demonstrate the necessity to determine adverse allelic variants of the known genes along with the classical deteminants of inherited thrombophilia in everyday obstetrical practice, as it allows to sportlight individual mechnisms of development of FLS and the most commonly seen complications of pregnancy.

Recombinant Thrombomodulin Safely Controls Disseminated Intravascular Coagulopathy in Acute Promyelocytic Leukemia – Results from the Japanese Pediatric Leukemia/Lymphoma Study Group AML-P05 Study

[Background] Acute promyelocytic leukemia (APL) cells induce dysregulation of coagulation and fibrinolysis, and most of the cases are complicated with disseminated intravascular coagulopathy (DIC) at the time of diagnosis. Administration of all-trans retinoic acid (ATRA) rapidly corrects dysregulation and improves DIC. But in some cases, management of DIC with ATRA alone is very difficult because addition of chemotherapeutic agents may reactivate fibrinolytic state. Thus the control of DIC is still an issue for the initial management of APL even in the ATRA era. Recombinant thrombomodulin (rTM) improves DIC by inhibiting excessive coagulation through the activation of protein C cascade (Saito H, et al. J Thromb Haemost 2007), and was approved in Japan for the treatment of DIC induced by severe infection or hematological malignancy in 2008. However, because activated protein C also facilitate fibrinolytic pathway by inhibiting plasminogen activator inhibitor-I, administration of rTM may lead to severe hemorrhage complication in patients with APL. Therefore, we retrospectively analyzed its efficacy and safety in pediatric APL.[Object & Methods] This study included 45 cases; 42 who registered to the Japanese Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-P05 study (UMIN Clinical Trials Registry, # UMIN000000645) and responded to the questionnaire and 3 cases outside the study. Questionnaire regarding the use of rTM and efficacy and safety issues, such as “Worst DIC score according to the diagnostic criteria of the Japanese Ministry of Health and Welfare (worst DIC score)” and “Days until resolution of DIC score from diagnosis of DIC (DIC duration)” were investigated.[Results] Among 45 cases analyzed, 38 (84%) were diagnosed to complicate with DIC, and 9 (24%) among them used rTM by physician’s decision. In these 9 cases (rTM+ group), mean duration of rTM administration was 13.8 days (7–24 days). WBC count and FDP level at diagnosis of APL were not different between rTM+ group and rTM-NOT used group (rTM- group). In rTM+ group, mean value of “Worst DIC score” was 5.8 and mean “DIC duration” from diagnosis was 16.8 days, both of them were not statistically significant with those in rTM- group (5.4 and 13.0 days, respectively). However, median “DIC duration” from commencement of rTM (11.5 days) seems shorter than “DIC duration” from diagnosis of DIC in rTM- group (13.0 days), although the difference was not statistically significant. Regarding safety issues, there was no death or severe hemorrhage in rTM+ group, while 3 cases (10%) of severe hemorrhage due to uncontrolled DIC were reported in rTM- group, with fatal brain hemorrhage in 1 case (3%) among them. There was no significant difference of induction rate between rTM+ group (1 case, 11%) and rTM- group (3 cases, 10%), although 2 cases of ATRA-refractory APL in rTM+ group who were additionally treated with arsenic trioxide.[Conclusions] In this retrospective analysis investigating efficacy and safety of rTM in the management of pediatric APL-related DIC, there were no significant differences regarding the efficacy of rTM because of the limited cases of rTM+ group. However, it is notable that there was no severe hemorrhage or death in rTM+ group. rTM seems to be safely used for the management of pediatric APL-related DIC. DisclosuresNo relevant conflicts of interest to declare.

Myeloperoxidase deficiency ameliorates progression of chronic kidney disease in mice.

Myeloperoxidase (MPO) is an enzyme expressed in neutrophils and monocytes/macrophages. Beside its well-defined role in innate immune defence, it may also be responsible for tissue damage. To identify the role of MPO in the progression of chronic kidney disease (CKD), we investigated CKD in a model of renal ablation in MPO knockout and wild-type mice. CKD was induced by 5/6 nephrectomy. Mice were followed for 10 wk to evaluate the impact of MPO deficiency on renal morbidity. Renal ablation induced CKD in wild-type mice with increased plasma levels of MPO compared with controls. No difference was found between MPO-deficient and wild-type mice regarding albuminuria 1 wk after renal ablation, indicating similar acute responses to renal ablation. Over the next 10 wk, however, MPO-deficient mice developed significantly less albuminuria and glomerular injury than wild-type mice. This was accompanied by a significantly lower renal mRNA expression of the fibrosis marker genes plasminogen activator inhibitor-I, collagen type III, and collagen type IV as well as matrix metalloproteinase-2 and matrix metalloproteinase-9. MPO-deficient mice also developed less renal inflammation after renal ablation, as indicated by a lower infiltration of CD3-positive T cells and F4/80-positive monocytes/macrophages compared with wild-type mice. In vitro chemotaxis of monocyte/macrophages isolated from MPO-deficient mice was impaired compared with wild-type mice. No significant differences were observed for mortality and blood pressure after renal ablation. In conclusion, these results demonstrate that MPO deficiency ameliorates renal injury in the renal ablation model of CKD in mice.

Blockade of cysteine-rich protein 61 attenuates renal inflammation and fibrosis after ischemic kidney injury.

Emerging data have suggested that acute kidney injury (AKI) is often incompletely repaired and can lead to chronic kidney disease (CKD), which is characterized by tubulointerstitial inflammation and fibrosis. However, the underlying mechanisms linking AKI to CKD remain obscure. The present study aimed to investigate the role of cysteine-rich protein 61 (Cyr61) after unilateral kidney ischemia-reperfusion injury (IRI) in mice. After IRI, increased expression of Cyr61 was detected, predominately in the proximal tubular epithelium. This was confirmed by in vitro experiments, which showed that hypoxia stimulated Cyr61 expression in cultured proximal tubular epithelial cells. The proinflammatory property of Cyr61 was indicated by its ability to upregulate monocyte chemoattractant protein-1 and IL-6. Additionally, we found elevated urinary Cyr61 excretion in patients with AKI. Notably, treatment of mice with an anti-Cyr61 antibody attenuated the upregulation of kidney monocyte chemoattractant protein-1, IL-6, IL-1β, and macrophage inflammatory protein-2 and reduced the infiltration of F4/80-positive macrophages on days 7 and 14 after IRI. In addition, blockade of Cyr61 reduced the mRNA expression of collagen, transforming growth factor-β, and plasminogen activator inhibitor-I as well as the degree of collagen fibril accumulation, as evaluated by picrosirius red staining, and levels of α-smooth muscle actin proteins by day 14. Concurrently, in the treated group, peritubular microvascular density was more preserved on day 14. We conclude that Cyr61 blockade inhibits the triad of inflammation, interstitial fibrosis, and capillary rarefaction after severe ischemic AKI. The results of this study expand the knowledge of the mechanisms underlying the AKI-to-CKD transition and suggest that Cyr61 is a potential therapeutic target.

Effect of Human Wharton's Jelly Mesenchymal Stem Cell Paracrine Signaling on Keloid Fibroblasts

Keloid scars are abnormal benign fibroproliferative tumors with high recurrence rates and no current efficacious treatment. Accumulating evidence suggests that human umbilical cord Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) have antifibrotic properties. Paracrine signaling is considered one of the main underlying mechanisms behind the therapeutic effects of mesenchymal stem cells. However, the paracrine signaling effects of WJ-MSCs on keloids have not yet been reported. The aim of this study is to investigate paracrine signaling effects of human WJ-MSCs on keloid fibroblasts in vitro. Human umbilical cords and keloid skin samples were obtained, and WJ-MSCs and keloid fibroblasts were isolated and cultured. One-way and two-way paracrine culture systems between both cell types were investigated. Plasminogen activator inhibitor-I and transforming growth factor-β2 (TGF-β2) transcripts were upregulated in keloid fibroblasts cultured with WJ-MSC-conditioned medium (WJ-MSC-CM) and cocultured with inserts, while showing lower TGF-β3 gene expression. Interleukin (IL)-6, IL-8, TGF-β1, and TGF-β2 protein expression was also enhanced. The WJ-MSC-CM-treated keloid fibroblasts showed higher proliferation rates than their control keloid fibroblasts with no significant change in apoptosis rate or migration ability. In our culture conditions, the indirect application of WJ-MSCs on keloid fibroblasts may enhance their profibrotic phenotype.

Taming Neonatal Hypoxic–Ischemic Brain Injury by Intranasal Delivery of Plasminogen Activator Inhibitor-1

Plasminogen activator inhibitor-I (PAI-1), a ≈50-kDa serine protease inhibitor, markedly reduces the extravascular toxicity of tissue-type plasminogen activator in experimental hypoxic-ischemic (HI) brain injury of newborns. However, the current treatment with PAI-1 requires intracerebroventricle injection to cross the blood-brain barrier, which is an invasive procedure of limited clinical potential. Thus, we tested whether intranasal administration of PAI-1 can bypass blood-brain barrier and mitigate neonatal HI brain injury. Rat pups were subjected to HI, with or without lipopolysaccharide pre-exposure, followed by intranasal delivery of a stable-mutant form of PAI-1 (CPAI). Immunoblotting showed that CPAI sequentially entered the olfactory bulbs and cerebral cortex after intranasal delivery and reduced ≈75% of brain atrophy in HI or lipopolysaccharide-sensitized HI injury. Mechanistically, CPAI attenuated HI-induced plasminogen activators and lipopolysaccharide/HI-induced nuclear factor-κB signaling, neuroinflammation, and blood-brain barrier permeability. Intranasal delivery of CPAI is an effective treatment of experimental HI brain injury of newborns. Clinical application of this experimental therapy merits further investigation.

Obesity and thrombin-generation profiles in women with venous thromboembolism

Obesity is a known risk factor for venous and arterial thrombosis but the mechanisms are still unclear. In women, obesity is correlated with low-grade inflammation and recent data show that BMI is positively associated with thrombin generation. We explored the correlations between obesity, inflammation and thrombin generation in women with increased thrombotic risk by looking at a cohort of women with prior venous thrombosis. One hundred and fifty-six women age 18-65 years were enrolled at diagnosis of first venous thromboembolism (VTE). Plasma samples were obtained at least 3 weeks after cessation of anticoagulant treatment. Thrombin generation was determined with the calibrated automated thrombography (CAT) assay and the Innovance ETP assay. Thrombin generation started later but was more pronounced with higher endogenous thrombin generation potential (ETP) determined with CAT in patients with obesity. The Innovance ETP assay showed results consistent with CAT. Furthermore, patients with obesity had significantly higher levels of fibrinogen, C-reactive protein and plasminogen activator inhibitor-I (PAI-I) than patients without obesity. Increased levels of fibrinogen were the main determinant of the prolonged lag-time in patients with obesity whereas higher levels of prothrombin could account for the difference in the ETP between the groups. We found an association between BMI and ETP values using two different methods to measure thrombin generation. Obesity correlated with increased thrombin generation in women with VTE and the main determinants of this hypercoagulable state were increased levels of fibrinogen and prothrombin. This shows a possible link between obesity, low-grade inflammation and increased thrombin generation in women at increased risk for future thrombosis.

Effects of Acute and One-Week Fatty Acid Lowering on Cardiac Function and Insulin Sensitivity in Relation with Myocardial and Muscle Fat and Adiponectin Levels

We tested the hypothesis that a persistent reduction in free fatty acid (FFA) levels improves cardiac function and systemic insulin sensitivity via a reduction in the myocardial and skeletal muscle adiposities and a modulation in adipokine release. Study subjects (body mass index 22-30 kg/m(2), 57 ± 3 yr old) underwent magnetic resonance imaging and spectroscopy to measure the cardiac function and the amounts of fat inside and around the myocardium and skeletal muscle, before (n = 10) and after acute (n = 8) and 1 wk (n = 7, one excluded from analysis) lowering of circulating FFA by acipimox. Circulating adipokines (leptin, adiponectin, resistin, TNFα, IL-6, IL-8, plasminogen activator inhibitor-I, macrophage chemoattractant protein-1) were measured. The ejection fraction (62 ± 2 vs. 56 ± 1%, P = 0.0035), cardiac output (6.6 ± 0.3 vs. 5.5 ± 0.2 liters/min, P = 0.0018), and forward work (708 ± 49 vs. 539 ± 44 mm Hg × liters/min, P = 0.018) were significantly lower after 1 wk of FFA lowering. In the six subjects undergoing all sessions, the stroke and end-diastolic volumes were also reduced, insulin sensitivity was increased by 33%, and adiponectinemia was decreased (-26%, P = 0.03). No change in intracellular cardiac and skeletal muscle triglyceride levels was observed. Metabolic changes correlated with the lowering of FFA. The reduction in cardiac function was related with changes in glycemia and insulin sensitivity, whereas the deflection in left ventricular work was correlated with the decline in FFA, lipid, and blood pressure levels. A 1-wk FFA depletion suppressed cardiac function and improved insulin sensitivity. Intracellular triglyceride deposits in the heart and skeletal muscle played no role in the observed changes. Our data show that FFA participate in the physiological regulation of adipokine levels.

Effects of eletro-acupuncturing at Zusanli point on coagulation and fibrinolysis in septic rats

Objective To investigate the effect of the electrical stimulation Zusanli (ST36) on coagulation and fibrinolysis in septic rats.Methods 192 male SD rats were randomly divided into six groups:saline group ( SHAME ),lipopolysaccharide ( LPS ) group,non-acupuncture stimulation group (non-EA),electrical stimulation Zusanli group (EA),vagotomy group (VGX),bungarotoxin group ( α-BGT).Six animals of every four time points [ before injection of LPS (0),2,4 and 6 h after injection]were killed after abdominal aortic blood was collected.The plasma was detected by enzyme linked immunosorbent assay (ELISA) for tumor necrosis factor-α (TNF-α),D-dimer,plasminogen activator inhibitor-Ⅰ (PAI-I) and tissue-type plasminogen activator (t-PA) levels,and the chromogenic substrate assay for antithrombin Ⅲ( AT-Ⅲ ) activity.Results At the 2nd h after injection of LPS,as compared with EA group (268.64 ±5.48) ng/L,the levels of plasma TNF-α in LPS group (332.71 ±8.23) ng/L and VGX group (449.70 ± 13.74) ng/L,and α-BGT group (448.89 ± 16.50) ng/L were significantly increased (P ＜0.05) ; At the 4th h,as compared with EA group (487.21 ±5.42) ng/L,the levels of plasma PAI in LPS group (681.27 ± 7.72) ng/L and VGX group (776.29 ± 3.23 ) ng/L,and e-BGT group (769.97 ± 5.50)ng/L were significantly increased ( P ＜ 0.05) ; At the 2nd h after injection of LPS,as compared with EA group (10.83 ±0.12) ng/L,the levels of plasma t-PA in LPS group (22.51 ± 0.47) ng/L and VGX group (27.10 ± 0.30) ng/L,and α-BGT group (27.01 ± 0.57 ) ng/L were significantly increased (P ＜0.05) ; At the 2nd h after injection of LPS,as compared with EA group (29.47 ± 3.58) ng/L,the levels of plasma D-dimer in LPS group (50.94 ±3.33) ng/L and VGX group (73.48 ±4.45) ng/L,and α-BGT group ( 70.10 ± 2.80 ) ng/L were significantly increased ( P ＜ 0.05 ) ; as compared with EA group (426.78 ±3.07) ng/L,the levels of plasma AT-Ⅲ in LPS group (368.76 ± 13.83) ng/L and VGX group (244.69 ±4.88 ) ng/L,and α-BGT group (241.85 ± 5.51 ) ng/L were significantly increased (P ＜0.05).Conclusion Electrical stimulation Zusanli can not only inhibit plasma levels of TNF-α but also can reduce inflammation,coagulation and fibrinolytic dysfunction in septic rats. Key words: Sepsis; Zusanli; Coagulation; Fibrinolysis; Electro-acupuncturing

Increased fibrinolysis increases bleeding in orthopedic patients receiving prophylactic fondaparinux

We evaluated hemostatic markers in patients who underwent major orthopedic surgery, including total hip and total knee arthroplasty, and were treated for the prophylaxis of deep vein thrombosis (DVT) with or without fondaparinux (anti-Xa group, n = 98 and without anti-Xa group, n = 20). The frequency of DVT was significantly higher in the without anti-Xa group than in the anti-Xa group, but the reduction of hemoglobin and fibrinolytic marker levels was significantly lower in the without anti-Xa group than in the anti-Xa group. Eighteen patients in the anti-Xa group showed a reduction in hemoglobin of more than 2 g/dl, and those individuals were considered to be the increased bleeding (IB) group. The concentration of fibrinolytic markers in the anti-Xa group was significantly higher in the IB group than in the non-IB group. There were also no significant differences in the levels of anti-Xa activity, plasminogen activator inhibitor-I, soluble fibrin and antithrombin between the IB and non-IB groups. In conclusion, elevated fibrinolysis induced by increased bleeding may lead to further increases in bleeding in patients receiving thromboprophylaxis with fondaparinux following major orthopedic surgery.

Haemostatic variables during normal menstrual cycle

For a number of haemostatic factors menstrual cycle variation has been studied. Such variation could have clinical implications for the timing of haemostatic testing in women. It was our objective to systematically review the literature about evidence for timing of haemostatic testing during menstrual cycle.We searched MEDLINE, EMBASE and the Cochrane library to identify studies that measured haemostatic variables [platelet function, von Willebrand factor (VWF), factor VIII (FVIII), factor IX (FIX), factor XI (FXI), factor XIII (FXIII), D-dimer, plasminogen activator inhibitor-I (PAI-I), tissue plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), α2-antiplasmin and fibrinogen] during normal menstrual cycle without hormonal contraceptives. Two investigators independently selected studies, and abstracted data in duplicate. We identified 1,046 studies of which we included 30 studies (25 longitudinal and 5 cross-sectional studies). All studies reported on haemostatic variables during menstrual cycle. Overall, most of the studies found no cyclic variation in VWF, FVIII, FXI, FXIII, fibrinolytic factors (PAI, t-PA, uPA, D-dimer and α2-antiplasmin) and fibrinogen. However, in studies where these variables showed any variation, they reached the lowest levels during menstrual and early follicular phase, especially for VWF, FVIII and platelet function tests. In conclusion, the optimal timing for haemostatic testing during menstrual cycle seems to be menstrual and early follicular phase.