Extraction of insoluble soybean fiber by alternating ultrasonic/alkali and its improved superior physicochemical and functional properties.

Okara, as a by-product of soybean processing, is rich in insoluble dietary fiber (IDF), which is a carbohydrate polymer with various insoluble polysaccharides. Nowadays, the extraction of IDF with excellent functional properties has become a research hotspot. In this work, we further proposed an alternating alkali/ultrasound method for the efficient extraction of IDF. The sequential treatments of alkali (A-ISF), alkali-ultrasonic (AU-ISF), ultrasonic-alkali (UA-ISF), ultrasonic-alkali-ultrasonic (UAU-ISF) and alkali-ultrasonic-alkali (AUA-ISF) were applied to extract insoluble soybean fiber (ISF). FTIR and XRD results proved the typical structure of ISFs, and TGA results demonstrated the improved thermal stability of UAU-ISF and AUA-ISF. Chemical composition measurement showed that UAU-ISF and AUA-ISF exhibited higher cellulose content (>83 %). SEM results revealed that ultrasonic treatment led to a decomposition of okara matrix and significant porous structure in ISFs with an amplified collapse effect, resulting in an increase of the pore size of ISFs, and strengthening the properties of UAU-ISF and AUA-ISF in higher water (>15 g/g)/oil (>12 g/g) holding capacities, cholesterol binding capacity (>36 mg/g), and cation exchange capacity (>0.3 mmol/g), thus providing new insights for the preparation of ISF with high functional properties that are beneficial for human intestinal health.

Cardioprotective effect of anisodamine against ischemia/reperfusion injury through the mitochondrial ATP-sensitive potassium channel.

Previous clinical studies have shown that anisodamine could improve no-reflow phenomenon and prevent reperfusion arrhythmias, but whether this protective effect is related to the antagonism of the M-type cholinergic receptor or other potential mechanisms is uncertain. The aim of the present study was to investigate the role of the mitochondrial ATP-sensitive potassium channel (mitoK ATP ) in cardioprotective effect of anisodamine against ischemia/reperfusion injury. Anisodamine and 5- hydroxydecanoic acid were used to explore the relationship between anisodamine and mitoK ATP . Using a Langendorff isolated heart ischemia/reperfusion injury model, hemodynamic parameters and reperfusion ventricular arrhythmia were evaluated; in addition, changes in myocardial infarct size, cTnI from coronary effluent and myocardial ultrastructure, as well as ATP, MDA and SOD in myocardial tissues, were detected. In the hypoxia/reoxygenation injury model of neonatal rat cardiomyocyte, cTnI release in the culture medium and levels of ATP, MDA and SOD in cardiomyocytes and mitochondrial membrane potential, were analyzed. Overall, anisodamine could significantly improve the hemodynamic indexes of isolated rat heart injured by ischemia/reperfusion, reduce the occurrence of ventricular reperfusion arrhythmia and myocardial infarction area, and improve the ultrastructural damage of myocardium and mitochondria. The in vitro results demonstrated that anisodamine could improve mitochondrial energy metabolism, reduce oxidative stress and stabilize mitochondrial membrane potential. The cardioprotective effects were significantly inhibited by 5-hydroxydecanoic acid. In conclusion, this study suggests that the opening of mitoK ATP could play an important role in the protective effect of anisodamine against myocardial ischemia/reperfusion injury.

Safety and efficacy of intracoronary prourokinase administration in patients with high thrombus burden.

OBJECTIVES: The study was designed to evaluate the effect of low-dose intracoronary prourokinase administration immediately after thrombus aspiration in patients with ST-segment elevation myocardial infarction (STEMI) presenting with a serious thrombus burden. METHODS: Consecutive STEMI patients with high thrombus burden received thrombus aspiration during primary percutaneous coronary intervention (PCI) were randomly assigned to study group (intracoronary prourokinase administration) or control group (intracoronary 0.9% sodium chloride administration). The primary endpoint was complete ST-segment resolution (STR) at 90 min after primary PCI, and the secondary endpoints included angiographic myocardial perfusion indexes. RESULTS: Patients in study group had a higher incidence of complete STR and myocardial blush grade 3 compared with those in control group (56.52% vs. 38.89%, P = 0.017 and 57.61% vs. 38.89%, P = 0.041). The peak cardiac troponin I value and corrected thrombolysis in myocardial infarction frame count were significantly lower in study group (52.16 ± 24.67 ng/mL vs. 60.91 ± 28.81 ng/mL, P = 0.029; and 19.57 ± 9.05 vs. 22.91 ± 10.22, P = 0.020). A significant improvement in left ventricular ejection fraction and major adverse cardiac events (MACEs)-free survival was observed in study group (55.22 ± 10.50% vs. 52.18 ± 9.39%, P = 0.041; 10.87% vs. 22.22%, P = 0.039) at the 6-month follow-up. The bleeding complication was similar in both groups (17.39% vs. 12.22%, P = 0.327). CONCLUSIONS: In STEMI patients with high thrombus burden, low-dose prourokinase intracoronary administered immediately after thrombus aspiration improves myocardial perfusion, cardiac function, and MACEs-free survival with no significant increase in major bleeding.

Predictive value of soluble suppression of tumourigenicity 2 on myocardial reperfusion.

BACKGROUND: High baseline level of soluble suppression of tumourigenicity 2 (sST2) was an independent predictor of cardiovascular death and heart failure in ST-segment elevation myocardial infarction (STEMI). AIMS: To investigate the value of serum sST2 baseline levels in predicting myocardial reperfusion in patients with STEMI undergoing primary percutaneous coronary intervention (PPCI). METHODS: Consecutive STEMI patients who underwent PPCI within 12 h after the onset of chest pain were enrolled, and were divided into Thrombolysis In Myocardial Infarction (TIMI) myocardial perfusion grading (TMPG) 0/1/2 group and TMPG 3 group based on post-procedural TMPG. Baseline clinical characteristics, lesions and procedural characteristics were compared. Univariate logistic regression and multivariate linear logistic analysis were performed to identify independent predictors of impaired myocardial reperfusion (TMPG 0/1/2). Receiver-operating characteristics curve (ROC) analysis of sST2 was performed to identify the optimum cut-off value for predicting the myocardial reperfusion. RESULTS: A total of 121 patients was enrolled in this study. Univariate logistic regression analysis showed that Killip II-III, high levels of sST2 and brain natriuretic peptide were risk factors of TMPG 0/1/2. Multivariable logistic regression analysis revealed that sST2 was an independent predictor of impaired myocardial reperfusion (odds ratio 12.318, 95% confidence interval 4.567-33.220, P < 0.001). ROC curve analysis showed that the area under curve of sST2 was 0.849, and the best cut-off value was 2.003 ng/mL, with a sensitivity of 89.2% and a specificity of 67.9%. CONCLUSION: The elevated levels of sST2 on admission were associated with impaired myocardial reperfusion in STEMI patients undergoing PPCI.

Beneficial effect of ticagrelor on microvascular perfusion in patients with ST-segment elevation myocardial infarction undergoing a primary percutaneous coronary intervention.

BACKGROUND: Ticagrelor significantly reduced the incidence of death, myocardial infarction, and stent thrombosis in patients with ST-segment elevation myocardial infarction (STEMI) intended for reperfusion with a primary percutaneous coronary intervention (pPCI). However, the effects of this drug on microvascular perfusion in patients presenting with STEMI have not been evaluated completely. PATIENTS AND METHODS: A total of 298 patients presenting with STEMI were randomized to either ticagrelor 180 mg loading, followed by 90 mg twice daily, or clopidogrel 600 mg loading, followed by 75 mg daily. The primary endpoint was ST-segment resolution at 90 min after pPCI. The secondary endpoints included myocardial blush grade and corrected thrombolysis in myocardial infarction frame count after the procedure. Left ventricular ejection fraction and major adverse cardiac events (MACE) at the 1- and 6-month follow-up time points were also recorded. RESULTS: There were no significant differences between the two groups with respect to baseline characteristics. Ticagrelor administration resulted in a higher rate of completed ST-segment resolution (58.67 vs. 39.86%, P=0.001), higher myocardial blush grade (2.63±0.64 vs. 2.41±0.71, P=0.005), and lower corrected thrombolysis in myocardial infarction frame count (19.68±7.38 vs. 22.35±8.30, P=0.004). At 6 months, left ventricular ejection fraction was higher (55.01±8.44 vs. 52.34±9.05%, P=0.009) in the ticagrelor group. Kaplan-Meier analysis showed that MACE-free survival had also improved in the ticagrelor group during the 1- and 6-month follow-up time points. CONCLUSION: Compared with clopidogrel, ticagrelor improves myocardial perfusion and left ventricular ejection fraction, and reduces the incidence of MACE for STEMI patients undergoing pPCI, with no significant increase in major bleeding.

Clinical efficacy of ticagrelor in patients undergoing emergency intervention for acute myocardial infarction and its impact on platelet aggregation rate.

OBJECTIVE: This study aims to investigate the clinical efficacy of ticagrelor in patients who underwent emergency percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI), and its impact on platelet aggregation rate. METHODS: A total of 257 AMI patients who underwent emergency PCI in our hospital were included in the present study. These patients were randomly divided into two groups: ticagrelor group (n = 129), patients took 180 mg of ticagrelor (qd) before the intervention, and subsequently took 90 mg of ticagrelor (bid) for maintenance; clopidogrel group (n = 128), patients took 300 mg of clopidogrel (qd) before PCI, and subsequently took 75 mg of clopidogrel (qd) for maintenance. Patients in both groups took 100 mg of aspirin. The major adverse cardiovascular events (MACE) within one year, changes in LVEF and LVEDD, platelet aggregation rate and drug safety before PCI and at one week and 30 days after PCI were observed in these two groups. RESULTS: The differences in baseline data between these two groups were not statistically significant. Within one year after the intervention, in the ticagrelor group, the total incidence of MACE was lower (P < 0.05), LVEF and LVEDD was improved (P < 0.05), and the decrease in platelet aggregation rate after the intervention was more significant (P < 0.05). Furthermore, the incidence of bleeding events was higher in the ticagrelor group than in the clopidogrel group (P < 0.05). CONCLUSIONS: Compared with clopidogrel, ticagrelor decreases the incidence of adverse cardiovascular events in AMI patients who underwent emergency PCI, does better in improving the fluctuation level of LVEF and LVEDD, and strongly inhibits platelet aggregation. Some patients encountered adverse drug events, but drug withdrawal or medication change did not occur.

Effect of Long-Term Administration of Nicorandil on Endothelial Function of the Radial Artery in Patients With Angina Undergoing Transradial Percutaneous Coronary Intervention.

We evaluated the effect of long-term administration of nicorandil on endothelial function of the radial artery in patients with angina undergoing elective transradial coronary intervention (TRI). A total of 127 patients were randomly assigned to nicorandil (standard medication plus nicorandil 5 mg twice daily, n = 64) or control group (standard medication except nicorandil, n = 63) immediately after TRI procedure. Radial artery diameter (RAD), flow-mediated dilation (FMD), and nitroglycerin-mediated dilation (NMD) of radial artery were measured 1 day before TRI as well as 1 day and 3 months after TRI by Ultrasound-Doppler. No significant difference was observed in the baseline RAD, FMD, and NMD between the 2 groups (all P > .05). In cannulated arm, at 3-month follow-up, RAD in nicorandil group was much larger than that in the control group (2.78 ± 0.27 mm vs 2.61 ± 0.30 mm, P = .001). Besides, FMD and NMD were much higher in nicorandil group than those in the control group (10.38% ± 2.43% vs 6.81% ± 1.86%; 15.94% ± 6.28% vs 10.46% ± 5.37%, respectively, all Ps < .001). In conclusion, long-term administration of nicorandil after TRI could improve the endothelial function of the cannulated radial artery.

Influence of Puncture Site on Radial Artery Occlusion After Transradial Coronary Intervention.

BACKGROUND: The risk of radial artery occlusion (RAO) needs particular attention in transradial intervention (TRI). Therefore, reducing vascular occlusion has an important clinical significance. The aim of this study was to determine the appropriate puncture site during TRI through comparing the occurrence of RAO between the different puncture sites to reduce the occurrence of RAO after TRI. METHODS: We prospectively assessed the occurrence of RAO in 606 consecutive patients undergoing TRI. Artery occlusion was evaluated with Doppler ultrasound in 2 days and 1 year after the intervention. Risk factors for RAO were evaluated using a multivariate model analysis. RESULTS: Of the 606 patients, the RAO occurred in 56 patients. Compared with TRI at 2-5 cm away from the radius styloid process, the odds ratio (OR) for occlusion risk at 0 cm and 1 cm were 9.65 (P = 0.033) and 8.90 (P = 0.040), respectively. The RAO occurred in the ratio of the arterial diameter to the sheath diameter ≤1 (OR = 2.45, P = 0.004). CONCLUSION: Distal puncture sites (0-1 cm away from the radius styloid process) can lead to a higher rate of RAO. TRIAL REGISTRATION: ClinicalTrials.gov, NCT01979627; https://clinicaltrials.gov/ct2/show/NCT01979627?term = NCT01979627 and rank = 1.

Intracoronary administration of different doses of anisodamine in primary percutaneous coronary intervention: protective effect in patients with ST-segment elevation myocardial infarction.

OBJECTIVE: The aim of this study was to evaluate the effects of intracoronary administration of anisodamine on myocardial reperfusion in patients with ST-segment elevation myocardial infarction (STEMI) undergoing a primary percutaneous coronary intervention (pPCI). METHODS: Patients with acute STEMI undergoing pPCI were enrolled in this randomized-controlled study (January 2014-June 2015) and divided randomly into four groups: group A (normal saline), group B (1000 µg anisodamine), group C (2000 µg anisodamine), and group D (4000 µg anisodamine). RESULTS: The study group included 140 patients. Percentages of thrombolysis in myocardial infarction (TIMI) myocardial perfusion grade 3, increased values of TIMI myocardial perfusion grade after stenting, and decreased values of corrected TIMI frame count in groups B, C, and D were all significantly higher than those in group A (P=0.031, 0.027, 0.003, and P<0.001, respectively). TIMI frame count after stenting in groups B, C, and D was significantly lower than that in group A (P=0.001). Left ventricular ejection fraction at 1 week after pPCI and at the 3-month follow-up, as well as the major adverse cardiac event-free survival rate in groups B, C, and D were higher than those in group A (P=0.027, 0.016, and 0.019, respectively). CONCLUSION: Intracoronary administration of anisodamine at different doses improved myocardial reperfusion in patients with STEMI undergoing pPCI and reduced major adverse cardiac events. The protective effect of anisodamine at a dose of 4000 µg might be better than the doses at 1000 and 2000 µg.

Intracoronary administration of anisodamine and nicorandil in individuals undergoing primary percutaneous coronary intervention for acute inferior myocardial infarction: A randomized factorial trial.

In acute inferior myocardial infarction (AIMI), numerous conventional drugs that are used to improve the myocardial microcirculation can significantly reduce blood pressure (BP) and coronary perfusion pressure, aggravate bradyarrhythmia and cause a deterioration in the hemodynamic state of the whole body, which greatly limits the application of these drugs in clinical settings. The aim of the present study was to assess the effect of anisodamine and nicorandil regimens on the prevention of no-reflow (NR) and the amelioration of myocardial reperfusion in patients with AIMI undergoing primary percutaneous coronary intervention (PCI). A total of 104 consecutive patients with AIMI were included in this study and randomly assigned to one of four groups: A (control group), B (anisodamine group), C (nicorandil group) and D (anisodamine and nicorandil group). Patients underwent PCI via transradial artery access and the angiographic results were evaluated. Coronary diastolic BP (DBP) and systolic and mean BPs were measured by invasive catheterization. The primary end-point was a post-PCI Thrombolysis In Myocardial Infarction (TIMI) myocardial perfusion grade (TMPG) of 3. Composite end-points (mortality + new MI + target vessel revascularization) were evaluated during the hospital stay and 30 days after discharge. Following the procedure, the proportion of patients achieving TMPG 3 was significantly higher in group D than that in the other groups (P=0.014); furthermore, the incidence of a postprocedural TIMI score of 3 was the highest in group D. Three days after the procedure, the peak creatine kinase-MB and cardiac troponin I levels were the lowest and the left ventricular ejection fraction was the highest in group D. A thrombus score of 3/4 and low DBP were the independent risk factors for poor myocardial reperfusion (expressed as TMPG <3), while 2 mg anisodamine plus 2 mg nicorandil prior to PCI was protective for optimal myocardial reperfusion following the procedure. The combination of anisodamine and nicorandil can effectively ameliorate myocardial reperfusion and protect cardiac function in patients with AIMI undergoing primary PCI.

Interleukin-1 Receptor-Associated Kinase 1/4 as a Novel Target for Inhibiting Neointimal Formation After Carotid Balloon Injury.

AIM: Interleukin-1 receptor-associated kinase 1 (IRAK1) and IRAK4 play essential roles in the induction of inflammatory gene products. We aimed to investigate the effect of the inhibition of IRAK1 and IRAK4 kinase activities on neointimal formation in rats with carotid artery balloon injuries using the IRAK1/4 inhibitor N-(2-Morpholinylethyl)-2-(3-nitrobenzoylamido)-benzimidazole, a cell-permeable benzimidazole compound. METHODS: Wistar rats and vascular smooth muscle cells (VSMCs) isolated from the thoracic aortas were used. Toll-like receptor 4 (TLR4)-mediated nuclear factor kappa B (NFκB) signaling pathway was revealed by microarrays analysis. In addition, the differential expression of the TLR4 pathway genes, including TLR4, IRAK1, IκBα, and interleukin-1ß (IL-1ß), was confirmed by quantitative real-time polymerase chain reaction. Immunohistochemical staining, elastic-van Gieson and Masson staining, 5-ethynyl-2´-deoxyuridine staining, enzyme-linked immunosorbent assay, transwell migration assay and western blotting were also contributed for relevant detection. RESULTS: The expression of TLR4 protein gradually increased at days 1, 3, 7, and 21 after balloon injury compared with the uninjured group. The dual inhibition of IRAK1 and IRAK4 attenuated neointimal formation and fibrotic remodeling after injury in vivo and suppressed VSMC proliferation and migration in vitro. The production of mediators such as tumor necrosis factor-α and IL-1ß in injured arteries were also reduced by the inhibition of IRAK1 and IRAK4. The expression of NFκB p65- and F4/80-positive cells in inhibitor rats were fewer than those in control rats at day 7, while IRAK1 expression was markedly higher at day 3 in inhibitor rats. Furthermore, western blotting analysis revealed that the IRAK1/4 inhibitor suppressed the IRAK1 and IRAK4 kinase activities and the activation of the TLR4-mediated NFκB pathway in vivo and in vitro. CONCLUSIONS: This study suggested that IRAK1/4 could serve as a potential therapeutic target to suppress neointimal formation in carotid arteries after balloon injury through the TLR4/NFκB signaling pathway.

Effects of liposomal prostaglandin E1 on periprocedural myocardial injury in patients with unstable angina undergoing an elective percutaneous coronary intervention.

OBJECTIVES: The aim of this study was to explore whether intravenous administration of liposomal prostaglandin E1 (lipo-PGE1) can reduce the incidence of periprocedural myocardial injury (PMI) in patients with unstable angina undergoing an elective percutaneous coronary intervention (PCI). PATIENTS AND METHODS: In this randomized-controlled study, a total of 219 patients were randomly assigned to a lipo-PGE1 group (n=110) and a control group (n=109). Patients in the lipo-PGE1 group received 20 µg/day of lipo-PGE1 diluted in 10 ml of normal saline through an intravenous injection over 5 min starting at 3 days before PCI and continuing for 4 days after PCI. In the control group, 10 ml of normal saline was administered using the same method. The primary end point was the occurrence of PMI defined as an elevation of cardiac troponin I above the upper limit of normal within 24 h after the procedure. The secondary end points were (i) changes in inflammatory factors including plasma high-sensitivity C-reactive protein, tumor necrosis factor α, and interleukin 6 before and at 24 h after PCI; (ii) the incidence of major adverse cardiac events in the patients during hospitalization and 30 days of follow-up after discharge, including cardiac deaths, severe heart failure, malignant arrhythmias, and target vessel revascularization. RESULTS: Within 24 h after PCI, the incidence of PMI was significantly lower in the lipo-PGE1 group compared with that in the control group (20 vs. 36.69%, P=0.009). Although the procedure induced a significant increase in high-sensitivity C-reactive protein, tumor necrosis factor α, and interleukin 6 levels, the values were significantly lower in the lipo-PGE1 group than those in the control group at 24 h after PCI (P<0.05). The proportion of thrombolysis in myocardial infarction grade 3 in the lipo-PGE1 group was higher than that in the control group (92.72 vs. 82.56%, P=0.037). There were no significant differences between the lipo-PGE1 group and the control group in the incidence of major adverse cardiac events during hospitalization and 30 days of follow-up (2.1 vs. 4%, P=0.72). Multivariate logistic analysis showed that lipo-PGE1 was an independent protective factor against PMI (odds ratio 0.385, 95% confidence interval 0.195-0.760, P=0.006). CONCLUSION: Intravenous lipo-PGE1 can reduce the incidence of PMI following elective PCI in patients with unstable angina. The benefit of lipo-PGE1 may be associated with the effects of anti-inflammation as well as improvement in coronary microvascular perfusion.

Effect of Intravenous Administration of Liposomal Prostaglandin E1 on Microcirculation in Patients with ST Elevation Myocardial Infarction Undergoing Primary Percutaneous Intervention.

BACKGROUND: Several studies have demonstrated that primary percutaneous coronary intervention (PCI) can result in reperfusion injury. This study aims to investigate the effectiveness of liposomal prostaglandin E1 (Lipo-PGE1, Alprostadil, Beijing Tide Pharmaceutical Co., Ltd.) for enhancing microcirculation in reperfusion injury. In addition, this study determined the optimal administration method for acute ST elevation myocardial infarction (STEMI) patients undergoing primary PCI. METHODS: Totally, 68 patients with STEMI were randomly assigned to two groups: intravenous administration of Lipo-PGE1 (Group A), and no Lipo-PGE1 administration (Group B). The corrected thrombolysis in myocardial infarction (TIMI) frame count (cTFC) and myocardial blush grade (MBG) were calculated. Patients were followed up for 6 months. Major adverse cardiac events (MACE) were also measured. RESULTS: There was no significant difference in the baseline characteristics between the two groups. The cTFC parameter in Group A was significantly lower than Group B (18.06 ± 2.06 vs. 25.31 ± 2.59, P < 0.01). The ratio of final MBG grade-3 was significantly higher (P < 0.05) in Group A (87.9%) relative to Group B (65.7%). There was no significant difference between the two groups in final TIMI-3 flow and no-reflow. Patients were followed up for 6 months, and the occurrence of MACE in Group A was significantly lower than that in Group B (6.1% vs. 25.9% respectively, P < 0.05). CONCLUSIONS: Myocardial microcirculation of reperfusion injury in patients with STEMI, after primary PCI, can be improved by administering Lipo-PGE1.

Cardioprotective Effect of Anisodamine Against Myocardial Ischemia Injury and its Influence on Cardiomyocytes Apoptosis.

Anisodamine is an ancient Chinese medicine derived from Tibet as a belladonna alkaloid, which is usually used for improvement of blood circulation in patients with organ phosphorus poisoning or shock. In this study, for the first time, we report its cardioprotective effects on preventing ischemia/reperfusion (I/R) injury of patients with acute myocardial infarction (AMI), and decreasing the myocardial infarction area and severity in heart of Sprague-Dawley (SD) rats. Our results suggest a potential molecular mechanism of anisodamine against the I/R injury in cardiomyocytes is associated with its anti-apoptotic effect. Anisodamine treatment decreases the expression of caspase-3 and caspase-8, and increases Bcl-2/Bax ratio in cardiomyocytes. Our data suggest that anisodamine can provide significant cardioprotection against I/R injury, potentially through the suppression of cardiomyocytes apoptosis.

Detecting DNA synthesis of neointimal formation after catheter balloon injury in GK and in Wistar rats: using 5-ethynyl-2'-deoxyuridine.

BACKGROUND: Neointimal formation plays an important role in the pathogenesis of coronary restenosis after percutaneous coronary intervention (PCI), especially in patients with diabetes mellitus. Recently, some studies have shown that 5-ethynyl-2'-deoxyuridine (EdU) incorporation can serve as a novel alternative to the 5-bromo-2'-deoxyuridine (BrdU) antibody detection method for detection of DNA synthesis in regenerating avian cochlea, chick embryo and the adult nervous system. However, few studies have been performed to assess the suitability of EdU for detecting DNA synthesis in vascular neointima. METHODS: The carotid artery balloon injury model was established in Goto-Kakizaki (GK) and Wistar rats. A Cell-LightTM EdU Kit was used to detect EdU-labeled cell nuclei of common carotid arteries at day 7 after catheter balloon injury. Different methods of injecting EdU were tested. The protein levels of proliferating cell nuclear antigen (PCNA) and p-Akt (Ser473), as well as the mRNA levels of PCNA were evaluated by Western blotting and quantitative real-time PCR (qRT-PCR), respectively. Immunohistochemical staining was also employed to visualize PCNA-positive cells. RESULTS: At day 7 after catheter balloon injury, far more EdU-positive and PCNA-positive cells were observed in GK rats. When comparing groups that received different EdU doses, it was found that the percentage of EdU-positive cells at a dose of 100 mg/kg body weight was than at doses of 25 mg/kg and 50 mg/kg. The number of positive cells was significantly higher in the repeated injection group compared to the single injection group. Further, after balloon injury DNA synthesis in GK rats was more notable than in Wistar rats. Neointimal formation in GK rats was more obvious than in Wistar rats. The protein levels of PCNA and p-Akt (Ser473) and the mRNA levels of PCNA were increased in injured rats as compared to uninjured rats, and were significantly higher in GK rats than in Wistar rats. CONCLUSION: By intraperitoneal injections of EdU at a dose of 100 mg/kg three times, EdU incorporation can detect carotid arterial DNA synthesis caused by neointimal formation in GK rats and Wistar rats at day 7 after balloon injury by the EdU click reaction quickly and effectively. Moreover, more obvious DNA synthesis in the vascular neointima could be observed in GK rats than in Wistar rats.