Impact of the COVID-19 Pandemic on ST-Elevation Myocardial Infarction Management in Hunan Province, China: A Multi-Center Observational Study.

Background: This study aimed to investigate the impact of the COVID-19 pandemic on ST-segment elevation myocardial infarction (STEMI) care in China. Methods: We conducted a multicenter, retrospective cohort study in Hunan province (adjacent to the epidemic center), China. Consecutive patients presenting with STEMI within 12 h of symptom onset and receiving primary percutaneous coronary intervention, pharmaco-invasive strategy and only thrombolytic treatment, were enrolled from January 23, 2020 to April 8, 2020 (COVID-19 era group). The same data were also collected for the equivalent period of 2019 (pre-COVID-19 era group). Results: A total of 610 patients with STEMI (COVID-19 era group n = 286, pre-COVID-19 era group n = 324) were included. There was a decline in the number of STEMI admissions by 10.5% and STEMI-related PCI procedures by 12.7% in 2020 compared with the equivalent period of 2019. The key time intervals including time from symptom onset to first medical contact, symptom onset to door, door-to-balloon, symptom onset to balloon and symptom onset to thrombolysis showed no significant difference between these two groups. There were no significant differences for in-hospital death and major adverse cardiovascular events between these two groups. Conclusion: During the COVID-19 pandemic outbreak in China, we observed a decline in the number of STEMI admissions and STEMI-related PCI procedures. However, the key quality indicators of STEMI care were not significantly affected. Restructuring health services during the COVID-19 pandemic has not significantly adversely influenced the in-hospital outcomes.

[Clinical effect and safety of clopidogrel combined with aspirin in antithrombotic therapy for children with Kawasaki disease complicated by small/medium-sized coronary artery aneurysms].

OBJECTIVE: To study the clinical effect and safety of clopidogrel combined with aspirin in antithrombotic therapy for children with Kawasaki disease (KD) complicated by coronary artery aneurysm (CAA). METHODS: A total of 77 KD children who were diagnosed with multiple small/medium-sized CAAs by echocardiography between January 2013 and June 2018 were enrolled. They were randomly divided into observation group with 38 children (treated with clopidogrel and aspirin) and control group with 39 children (treated with low-molecular-weight heparin and aspirin). All children were followed up regularly, and the first 3 months of the course of the disease was the observation period. The children were observed in terms of the change of the coronary artery and the incidence of complications. RESULTS: At month 3 of follow-up, among the children in the observation group, 6 had normal coronary artery, 11 had coronary artery retraction, 19 had stable coronary artery, and 2 progressed to giant coronary aneurysm; among the children in the control group, 7 had normal coronary artery, 12 had coronary artery retraction, 19 had stable coronary artery, and 1 progressed to giant coronary aneurysm; there was no significant difference in the change of the coronary artery between the two groups (P>0.05). There were 2 cases of epistaxis and 6 cases of skin ecchymosis in the observation group, and 1 case of epistaxis and 7 cases of petechiae and ecchymosis at the injection site in the control group, and no other serious bleeding events were observed in either group. CONCLUSIONS: Clopidogrel combined with low-dose aspirin is safe and effective in antithrombotic therapy for children with KD complicated by CAA.

LncRNA-ANRIL inhibits cell senescence of vascular smooth muscle cells by regulating miR-181a/Sirt1.

BACKGROUND: Cardiovascular disease is one of the major threats to human life and health, and vascular aging is an important cause of its occurrence. Antisense non-coding RNA in the INK4 locus (ANRIL) is a kind of long non-coding RNA (lncRNA) that plays important roles in cell senescence. However, the role and mechanism of ANRIL in senescence of vascular smooth muscle cells (VSMCs) are unclear. METHODS: Cell viability and cell cycle were evaluated using an MTT assay and flow cytometry analysis, respectively. Senescence-associated (SA)-ß-galactosidase (gal) staining was used to determine cell senescence. Dual luciferase reporter assays were conducted to confirm the binding of ANRIL and miR-181a, as well as miR-181a and Sirt1. The expression of ANRIL, miR-181a, and Sirt1 was determined using qRT-PCR and protein levels of SA-ß-gal and p53-p21 pathway-related proteins were evaluated by Western blotting. RESULTS: ANRIL and Sirt1 were down-regulated, whereas miR-181a was up-regulated in aging VSMCs. In young and aging VSMCs, over-expression of ANRIL could down-regulate miR-181a and up-regulate Sirt1. MTT and SA-ß-gal staining assays showed that over-expression of ANRIL and inhibition of miR-181a promoted cell viability and inhibited VSMC senescence. The dual-luciferase reporter assay determined that miR-181a directly targets ANRIL and the 3'-UTR of Sirt1. Furthermore, over-expression of ANRIL inhibited cell cycle arrest and the p53-p21 pathway. CONCLUSION: ANRIL promotes cell viability and inhibits senescence in VSMCs, possibly by regulating miR-181a/Sirt1, and alleviating cell cycle arrest by inhibiting the p53-p21 pathway. This study provides novel insights for the role of ANRIL in the development of cell senescence.

Construction of a novel vector expressing Survivin-shRNA and fusion suicide gene yCDglyTK and its application in inhibiting proliferation and migration of colon cancer cells.

Despite progress achieved in cancer chemotherapy in recent decades, adverse effects remain a limiting factor for a number of patients with colorectal cancer, suggesting the requirement for novel therapeutic strategies. Gene therapy appears to be a promising strategy for treating cancer. The present study aimed to investigate the anti-tumor effect of a combined gene therapy, using Survivin downregulation by RNAi and a fusion suicide gene yCDglyTK therapy system. A triple-gene vector expressing Survivin-targeted small hairpin RNA (Survivin-shRNA) and fusion suicide gene yCDglyTK was constructed, and administered to HCT116 cells. Survivin expression decreased significantly and yCDglyTK fusion gene expression was confirmed by both reverse transcription-quantitative polymerase chain reaction and western blot analysis. Introduction of Survivin-shRNA into yCDglyTK/prodrug system eradicated colon cancer cells and induced apoptosis more effectively. Furthermore, this therapeutic system is able to inhibit the migration of HCT116 cells. These results indicate that the recombinant plasmid may serve as a novel gene therapy approach to treat colorectal carcinoma.

Interferon-α-induced CD100 on naïve CD8+ T cells enhances antiviral responses to hepatitis C infection through CD72 signal transduction.

Objectives We investigated the effects of CD100 on naïve CD8+ T cells during hepatitis C virus (HCV) infection after interferon-α (IFN-α) therapy to clarify the mechanism underlying the effect of IFN-α in enhancing the antiviral response. Methods The CD100 molecules on subsets of CD8+ T cells were analysed with flow cytometry. The effects of CD100-overexpressing naïve CD8+ T cells were determined with ELISAs and an MTT cytotoxicity assay. The role of CD100-CD72 signal transduction was analysed with a neutralization and transwell assays. Results HCV infection reduced CD100 expression on CD8+ T cells, whereas IFN-α treatment significantly increased CD100 expression on naïve CD8+ T cells. The increased CD100 interacted with the CD72 receptor and enhanced PBMC cytokine secretion (IFN-γ and tumour necrosis factor-α) and cytotoxicity. Conclusions IFN-α-induced CD100 on naïve CD8+ T cells promotes PBMC cytokine secretion and cytotoxicity through CD100-CD72 signalling during HCV infection.

[Relationship between heart rate variability and coronary artery lesion in children with Kawasaki disease].

OBJECTIVE: To explore the correlation of heart rate variability (HRV) indices with cardiac troponin I (cTnI) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) in children with Kawasaki disease (KD) and their prognostic value. METHODS: A total of 130 children with KD were assigned into coronary artery lesion (CAL) group (n=47) and non-coronary artery lesion (NCAL) group (n=83). Meanwhile, 110 healthy children and 29 children in the recovery stage of non-cardiovascular diseases were selected as control and non-KD groups, respectively. Patients in the four groups received 24-hour HRV monitoring. Levels of serum cTnI and NT-proBNP were measured in the KD and the non-KD group. RESULTS: Compared with the controls of the same sex and age, the KD patients had significantly reduced standard deviation of all normal sinus RR intervals (SDNN), mean of SDNN (SDNN index), percentage of successive normal sinus RR intervals>50 ms (pNN50), very low frequency (VLF), low frequency (LF), and high frequency (HF) but a significantly increased LF/HF ratio (P<0.05). The HRV indices including SDNN, standard deviation of all mean 5-minute RR intervals (SDANN), SDNN index, root mean squared successive difference, pNN50, VLF, LF, and HF in the CAL group all significantly decreased compared with those in the control and non-KD groups, while the LF/HF ratio was higher in the CAL group than in the control group (P<0.05). The serum levels of cTnI and NT-proBNP in the CAL and NCAL groups were significantly higher than those in the non-KD group (P<0.05). In children with KD, serum cTnI level was negatively correlated with SDNN and HF but positively correlated with the LF/HF ratio (P<0.05); serum NT-proBNP level was negatively correlated with SDNN, SDANN, and HF (P<0.05). CONCLUSIONS: HRV indices have certain clinical significance in assessing CAL of children with KD.

microRNA-214 functions as a tumor suppressor in human colon cancer via the suppression of ADP-ribosylation factor-like protein 2.

microRNAs (miRNAs/miRs) are a conserved class of endogenous, short non-coding RNAs that post-transcriptionally regulate the expression of genes involved in diverse cellular processes. miR-214 has been reported to be associated with several cancers, including human colon cancer. However, the function of miR-214 in colon cancer development is poorly understood. In the current study, miR-214 was demonstrated to be downregulated in colon cancer tissues compared with healthy colon tissues. Functional studies showed that miR-214 overexpression results in the inhibition of cell viability, colony formation and proliferation, and the induction of cell apoptosis. ADP-ribosylation factor-like protein 2 (ARL2) is predicted to be a target candidate of miR-214. A luciferase reporter assay, western blot analysis and quantitative polymerase chain reaction were performed, which revealed that miR-214 negatively regulates ARL2 expression by targeting its 3' untranslated region directly. In conclusion, the results of the present study revealed that miR-214 suppresses colon cancer cell growth via the suppression of ARL2, and indicated that miR-214 may present a significant potential therapeutic target for colon cancer.

[The role of endogenous vascular elastase in coronary artery reconstruction in Kawasaki disease].

OBJECTIVE: To investigate the role of endogenous vascular elastase (EVE) in coronary artery between reconstruction among pediatric patients with Kawasaki disease (KD). METHODS: Sixty children who were diagnosed with KD between January 2012 and April 2013 were selected as the case group, and peripheral venous blood samples were collected on days 0-11 (pathological stage I) and days 12-25 (pathological stage II) after the onset of disease; another 60 children without KD who visited the hospital due to acute fever during the same period were selected as the control group, and fasting peripheral venous blood samples were collected in the acute stage of fever. For both groups, serum levels of EVE and interleukin-6 (IL-6) and plasma vascular endothelial growth factor (VEGF) level were measured by enzyme-linked immunosorbent assay. For the case group, ultrasonic cardiography was used to detect coronary artery lesions (CALs) at the first, second and fourth weekends. The correlations of EVE level with IL-6 and VEGF levels were evaluated by Pearson correlation analysis. RESULTS: Serum levels of EVE and IL-6 in the case group in pathological stages I and II were significantly higher than in the control group (P<0.05), but plasma VEGF levels in stages I and II were significantly lower than in the control group (P<0.05); in the case group, EVE and IL-6 levels were significantly higher in stage II than in stage I (P<0.05). In pathological stage II, KD patients with CALs had significantly higher serum levels of EVE and IL-6 but significantly lower plasma VEGF levels compared with those without CALs (P<0.05); KD patients with coronary artery aneurysms (CAAs) had significantly higher serum levels of EVE and IL-6 but significantly lower plasma VEGF level compared with those without CAAs (P<0.05 for all). EVE level was positively correlated with IL-6 level (r=0.915, P<0.05), yet negatively correlated with VEGF level (r=-0.769, P<0.05). CONCLUSIONS: EVE may participate in coronary artery reconstruction in children with KD. To interfere EVE activity may reduce and prevent CALs.

[Plasma concentration of growth-differentiation factor-15 in children with congenital heart disease: relation ship to heart function and diagnostic value in heart failure].

OBJECTIVE: To study the correlation between growth differentiation factor-15(GDF-15) and cardiac function in pediatric patients with congenital heart disease, and the diagnostic value of GDF-15 in heart failure(HF). METHODS: From March 2011 to May 2012, 97 pediatric patients with congenital heart disease(CHD) who consecutively attended Chengdu Women's & Children's Central Hospital were enrolled in the study and assigned to HF (patients with heart failure, n=71) and Non-HF(patients without heart failure, n=26) groups. HF was defined as patients presenting with modified Ross score≥3. Plasma concentrations of GDF-15 and NT-proBNP were determined using ELISA. Left ventricular ejection fraction(LVEF) was tested by echocardiography. The correlation between GDF-15 and modified Ross score, LVEF and NT-proBNP was evaluated with Spearman's analysis. The area under the receiver-operating characteristic(ROC) curve for GDF-15 was examined, and the cut-off concentration of GDF-15 for diagnosing HF was detected. RESULTS: The HF group demonstrated higher levels of GDF-15 and NT-proBNP, and a lower LVEF level (P<0.01) than the Non-HF group. Plasma GDF-15 level was positively correlated with modified Ross score and plasma NT-proBNP concentration (r=0.705, r=0.810 respectively; P<0.01), and negatively correlated with LVEF(r=-0.391, P<0.01). According to ROC analysis, the AUC of GDF-15 for detection of HF was 0.757. Sensitivity and specificity was 68.8% and 71.2% respectively for the cut-off value of 1306 ng/mL. CONCLUSIONS: Plasma GDF-15 levels are significantly elevated in children with HF induced by CHD. Plasma GDF-15 levels are related to cardiac function, LVEF and plasma concentration of NT-proBNP. GDF-15 may potentially indicate HF in pediatric patients with CHD.

[Risk factors for coronary artery lesions in children with Kawasaki disease].

OBJECTIVE: To study risk factors for the development of coronary artery lesions (CAL) in children with Kawasaki disease (KD). METHODS: The clinical data of 527 children with KD between January 2006 and January 2009 were retrospectively reviewed. A total of 15 potential factors associated with occurrence of CAL were evaluated by univariate analysis and multivariate logistic regression analysis. RESULTS: The univariate analysis showed that age, gender, KD type, starting time of intravenous immunoglobulin (IVIG) treatment, response to IVIG treatment, additional treatment with corticosteroids, duration of fever and serum C-reactive protein level were significantly different between patients with and without CAL (P<0.05). Multivariate logistic regression analysis showed that an age of less than 1 year (OR=2.076, P<0.05) or greater than 8 years (OR=1.890, P<0.05), male sex (OR=1.972, P<0.05), incomplete KD (OR=1.426, P<0.05), delayed starting time of IVIG treatment (10 days after onset) (OR=3.251, P<0.05), no response to IVIG (OR=2.301, P<0.05) and fever duration of more than 10 days (OR=1.694, P<0.05) were independent risk factors for the development of CAL, whereas early starting time of IVIG treatment (before 5 days after onset) was a protective factor (OR=0.248, P<0.05). CONCLUSIONS: The occurrence of CAL is associated with many factors in children with KD. Age of less than 1 year or greater than 8 years, male sex, incomplete KD, delayed IVIG treatment after onset, no response to IVIG treatment and prolonged fever duration have been identified as risk factors for the development of CAL.

[Characterizations of atrial contractions triggering paroxysmal atrial fibrillation and the effect of atrial fibrillation prevention pacing].

OBJECTIVE: To evaluate the electrocardiographic characterizations of atrial contractions(AC) triggering paroxysmal atrial fibrillation(AF), and to explore the effects of AF prevention pacing on their electrocardiographic characterizations. METHODS: Twenty-four patients with the implantation of AF therapy pacemaker(Vitatron 900E) were analyzed by AC triggering paroxysmal AF with Holter monitoring in the study. AC compluing interval, compensatory pause and frequency 2 minutes before the AF or during the AC were compared between the induced paroxysmal AF group and noinduced paroxysmal AF group, and the preventive effect of AF on the post-PAC response program was investigated. RESULTS: There was significant difference in the AC compluing interval [(352.3 +/-30.4) vs (421.8 42.5)ms], compensatory pause [(963 +/-109) vs (733 +/-124) ms], and frequency [(34.8 +/-18.9) vs (12.7 +/-8.7)/min] 2 minutes before the AF or during the AC in the induced paroxysmal AF group, compared with those in the noinduced paroxysmal AF group (all P<0.05). The AF of 7 patients were controlled by atrial overdrive pacing therapy, 17 patients by post-AC-response or/and post-exercise control therapy, 6 patients by the above therapy combining with cordarone (0.2g/d). CONCLUSION: AC triggering paroxysmal AF is related to the compluing interval, compensatory pause and frequency 2 minutes before the paroxysmal AF or during the AC, AF prevention pacing may be helpful for the paroxysmal AF induced by AC.

[Effect of atrial pacing on the sleep apnea syndrome in elderly patients].

OBJECTIVE: To evaluate the efficacy of atrial pacing to increase the heart rate during sleep in elderly patients with sleep apnea syndrome. METHODS: Sixteen elderly patients with central type or obstructive type sleep apnea received permanent atrial-synchronous ventricular pacemakers for symptomatic sinus bradycardia were analysed in this study. All patients received polysomnographic evaluations for 3 consecutive nights. All patients were evaluated at the base-line, and then were randomly divided into 2 groups at the first night. In the following 2 nights, one group was monitored in spontaneous rhythm model and the other in dual-chamber pacing model with atrial overdrive (increasing by 15 beats per minute on the basic rate of the mean nocturnal sinus), and then the two groups were switched at the third night. The total duration and number of episodes of sleep apnea or hypopnea were analyzed, and compared between the two models. RESULTS: The mean 24 h sinus rate in the spontaneous rhythm mode was 55+/-9 beats per minute at the base line, as compared in the 72+/-4 beats per minute in the atrial overdrive pacing model. There was statistic significant difference between the 2 models (P<0.05). There was no difference in the total duration of sleep between spontaneous rhythm model and atrial overdrive pacing model [(322+/-48) minutes vs (330+/-52) minutes, P>0.05 ]. The hypopnea index reduced from 9+/-3 in the spontaneous rhythm model to 3+/-3 in the atrial overdrive pacing model (P<0.01). The index of apnea and hypopnea was 28+/-21 in the spontaneous rhythm model, as compared with 10+/-13 in the atrial overdrive pacing model (P<0.01). CONCLUSION: Atrial overdrive pacing can significantly reduce the number of episodes of central type or obstructive type sleep apnea, but doesn't decrease the total sleep time in elderly patients with sleep apnea syndrome.

[Detecting the activity of antibodies induced by recombinant TGFbeta1 vaccine].

OBJECTIVE: To measure the neutralization activity in vitro of the antibodies induced by recombinant TGFbeta1 vaccine and to evaluate the vaccine's anti-liver fibrosis activity. METHODS: Balb/c mice were immunized with a fusion protein of the human TGFbeta1 epitope-inserted into a hepatitis B core antigen using a prokaryotic expression system. The antibody produced by the recombinant vaccine was determined using ELISA. The biological activity of the anti-TGFbeta1 antibody induced by the vaccine was measured by MTT using mink lung epithelial cell Mv-1-Lu as inhibiting cells. The fusion protein was used as a vaccine in a mice hepatic-fibrosis model. RESULTS: A high titer of anti-TGFbeta1 antibody and a low of anti-HBc antibody were detected in the mice after the immunization. The serum antibodies induced combined with the fusion and antigenic peptide prevented the TGFbeta1 inhibiting activity in the Mv-1-Lu cell. CONCLUSION: Recombinant fusion protein can be used as a cytokine vaccine to induce high titers of anti-TGFbeta1 antibodies. Our results show the potentiality of the fusion protein to be used as a vaccine for preventing liver fibrosis.

[Fusion of TGF-beta 1 antigenic determinant and HBV core antigen as an anti-TGF-beta 1 vaccine].

OBJECTIVES: To examine the expression and purification of the TGFbeta1 vaccine from prokaryotic expression system and to determine the antigenicity of the fusion protein of recombinant vector pET28a/ HBcAg1-71-TGFbeta132-HBcAg89-144. METHODS: The reconstructed vector pGEMEX-1/CTC was subcloned to pET28a and transformed into E. coli BL21 (DE3). The recombinant 6xHis- HBcAg1-71- TGFbeta132- HBcAg89-144 was to be expressed after induction by IPTG and purified with Ni-NTA-His affinity chromatography. The detection of the formation of core-like particles was done under an electron microscope and of their antigenity by using ELISA and Western blot. RESULTS: A 2.46 x 10(4) protein was obtained by optimizing the conditions for both expression and purification. The protein had the TGFbeta1 antigenicity but not a HBc antigenity and the formed core-like particles were bigger than natural core particles. CONCLUSION: The recombinant fusion protein in the prokaryotic expressed system can be used as an anti-TGFbeta1 vaccine to inhibit hepatic fibrosis.

Construction of prokaryotic expression system of TGF-beta1 epitope gene and identification of recombinant fusion protein immunity.

AIM: To insert the constructed TGF-beta(1) epitope gene into the el loop of C-terminus of truncated hepatitis B core antigen to increase TGF-beta(1) antigenicity in its prokaryotic expression system and to identify immunity of the expressed recombinant protein in order to exploit the possibility for obtaining anti-TGF-beta(1) vaccine. METHODS: The TGF-beta(1) encoding epitope gene (the mature TGF-beta(1) from 78-109 amino acid residues, TGF-beta(1)(32)) was amplified by polymerase chain reaction from the recombinant pGEM-7z/ TGF-beta(1)(32) vector. The HBcAg gene fragments (encoding HBcAg from 1-71 and 89-144 amino acid residues) were amplified from PYTA1-HBcAg vector. The recombinant vector pGEMEX-1 was used to insert HBcAg1-71, TGF-beta(1)(32) and HBcAg89-144 into restrictive endonuclease enzyme and ligated with T(4) ligase. The fusion gene fragments HBcAg1-71-TGF-beta(1)(32)- HBcAg89-144 were recloned to pET28a(+) and the DNA sequence was confirmed by the dideoxy chain termination method. The recombinant vector pET28a (+)/CTC was transformed and expressed in E. coli BL21 (DE3) under induction of IPTG. After purification with Ni(+2)-NTA agarose resins, the antigenicity of purified protein was detected by ELISA and Western blot and visualized under electron microscope. RESULTS: Enzyme digestion analysis and sequencing showed that TGF-beta(1) epitope gene was inserted into the el loop of C-terminus of truncated hepatitis B core antigen. SDS-PAGE analysis showed that relative molecular mass (Mr) of the expressed product by pET28a (+)/CTC was Mr 24,600. The output of the target recombinant protein was approximately 34.8% of the total bacterial protein, mainly presented in the form of inclusion body. Western blotting and ELISA demonstrated that the fusion protein could combine with anti-TGF-beta(1) polyclonal IgG but not with anti-HBcAg. The purity of protein was about 90% and the protein was in the form of self-assembling particles visualized under electron microscope. This fusion protein had good anti-TGF-beta(1) antigenicity and could be used as anti-TGF-beta(1) vaccine. CONCLUSION: A recombinant prokaryotic expression system with high expression efficiency of the target TGF-beta(1) epitope gene was successfully established. The fusion protein is in the form of self-assembling particles and HBcAg can increase the antigenicity of TGF-beta(1). The expressed TGF-beta(1) epitope gene shows good immunogenicity and antigenicity.