The lncRNA-miRNA-integrin alpha V ceRNA network can affect the occurrence and prognosis of gastric cancer.

OBJECTIVES: The aim of this study was to explore the role of integrin alpha V (ITGAV) and the related long noncoding RNA-microRNA-messenger RNA competing endogenous RNA (lncRNA-miRNA-mRNA ceRNA) network in the development and prognosis of cancers, especially gastric cancer (GC), through bioinformatic analysis. METHODS: Pan-cancer and GC data were collected from the UCSC Xena website, and validation datasets were obtained from the Gene Expression Omnibus (GEO). R (version 3.6.3), GraphPad Prism 8, and SPSS 23.0 software were used to analyze data and prepare figures. RESULTS: The expression of ITGAV in tumor tissues was higher than that of normal tissues in ten cancer types. A lower expression of ITGAV in five tumors (CESC, LGG, LIHC, MESO, and STAD) predicted better patient prognosis. In GC, the mRNA and protein expression of ITGAV in tumor tissues was higher than that of normal tissues. Patients with high ITGAV expression had poor prognosis and clinical characteristics, including worse grades and more advanced stages. Patients with higher ITGAV expression had higher immune and stromal scores and lower purity (P<0.05). In addition, seven miRNAs were found that were negatively correlated with ITGAV expression through the website; high expression of these miRNAs indicated a better prognosis. Using this correlation, the authors built the lncRNA-miRNA-ITGAV ceRNA network, to predict the prognosis of GC. CONCLUSIONS: This study showed that ITGAV could be considered a prognostic factor for GC, and an lncRNA-miRNA-ITGAV ceRNA network was built to promote the exploration of the mechanism and prognosis of GC.

del Nido cardioplegia better preserves cardiac diastolic function but histidine-tryptophan-ketoglutarate is better for endothelial function.

OBJECTIVES: The effectiveness of myocardial protection of cardioplegia has been a matter of debate for decades. This study was designed to compare cardiac and endothelial protection of 3 clinically used cardioplegias: del Nido cardioplegia (DNC), histidine-tryptophan-ketoglutarate (HTK) and blood cardioplegia (BC) followed by HTK (BC + HTK) in a rat model of ischaemia/reperfusion (I/R). METHODS: Sixty male Wistar rats were subjected to either 120 min of global ischaemia at 4°C followed by 90 min of reperfusion (I/R) at 37°C or no I/R (control) in a Langendorff apparatus and were randomly allocated to 5 groups: control, I/R, DNC, HTK and BC + HTK. Cold cardioplegia solutions were administered at doses of 20 ml/kg for DNC and HTK or 10 ml/kg for BC followed by HTK. Haemodynamic parameters were continuously recorded using an intraventricular balloon. The endothelium-dependent relaxation to acetylcholine was measured in the left anterior descending artery using a myograph. Protein expression of cardiac troponin T (cTnT) and creatine kinase MB was determined by western blot. RESULTS: During reperfusion, HTK had higher left ventricular systolic pressure whereas DNC had lower left ventricular end-diastolic pressure, better left ventricular developed pressure and best +dp/dtmax and -dp/dtmax than the other 2 groups but the differences disappeared at the end of the reperfusion. HTK or BC + HTK preserves the acetylcholine-induced endothelium-dependent relaxation better than DNC (Emax = 48.2 ± 8.0% in DNC vs 75.0 ± 8.0% in HTK, P < 0.05; vs 96.9 ± 3.5% in BC + HTK, P < 0.001). The protein levels of cTnT and creatine kinase MB were downregulated in the 3 groups. CONCLUSIONS: All 3 cardioplegias prevented myocardial damage against I/R injury at the end of reperfusion. DNC demonstrated better preserved diastolic function of the left ventricle whereas HTK or BC + HTK showed better preserved coronary endothelial function. These findings may suggest that currently no 'perfect' cardioplegia exists and that exploration for the 'perfect' cardioplegia is needed.

[Effect of staging treatment of Tongyuan acupuncture on pregnancy outcome in patients with recurrent implantation failure of thin endometrium type].

OBJECTIVE: To compare the effect of Tongyuan acupuncture combined with medication and medication alone on pregnancy outcome in patients with recurrent implantation failure (RIF) of thin endometrium type. METHODS: A total of 74 patients with RIF of thin endometrium type undergoing freeze-thaw embryo transfer were randomly divided into an observation group (37 cases) and a control group (37 cases). The patients in the control group were treated with freeze-thaw embryo transfer in hormone replacement cycle, and the estradiol valerate tablets were taken orally from the fifth day of menstruation, 2 mg per day. On the basis of the control group, the observation group was additionally treated with Tongyuan acupuncture at Baihui (GV 20), Dazhui (GV 14), Qihai (CV 6), Guanyuan (CV 4), etc., combined with other acupoints based on syndrome differentiation and menstrual stage, once every other day. Both groups were treated for 3 menstrual cycles. The clinical pregnancy rate and embryo implantation rate of the two groups were observed after transplantation; the endometrial thickness and type, resistance index (RI) and pulsatility index (PI) of endometrial blood flow were measured before treatment and one day before transplantation, and adverse reactions was recorded. RESULTS: The clinical pregnancy rate was 37.8% (14/37) in the observation group, which was higher than 16.2% (6/37) in the control group (P<0.05). There was no significant difference in embryo implantation rate between the two groups (P>0.05). One day before transplantation, the endometrial thickness and the proportion of type A in endometrial classification in the two groups were increased compared with those before treatment (P<0.01), and those in the observation group were higher than the control group (P<0.01, P<0.05). The PI and RI of endometrial blood flow in the two groups were lower than those before treatment (P<0.01), and those in the observation group were lower than the control group (P<0.01, P<0.05). During the treatment, 6 patients in the control group had discomfort such as breast distending pain, stomach pain, dizziness and nausea, and there were no adverse reaction in the observation group. CONCLUSION: On the basis of conventional medication, Tongyuan acupuncture could increase the endometrial thickness, improve endometrial receptivity, improve pregnancy outcome and reduce adverse reactions in patients with RIF of thin endometrial type.

Homocysteine alters vasoreactivity of human internal mammary artery by affecting the KCa channel family.

BACKGROUND: Hyperhomocysteinemia is an independent risk factor for atherosclerotic heart disease. We previously demonstrated that disruption of calcium-activated potassium (KCa) channel activity is involved in homocysteine-induced dilatory dysfunction of porcine coronary arteries. Recently we reported that the KCa channel family, including large-, intermediate-, and small-conductance KCa (BKCa, IKCa, and SKCa) subtypes, are abundantly expressed in human internal mammary artery (IMA). In this study, we further investigated whether homocysteine affects the expression and functionality of the KCa channel family in this commonly used graft for coronary artery bypass surgery (CABG). METHODS: Residual IMA segments obtained from patients undergoing CABG were studied in a myograph for the role of KCa subtypes in both vasorelaxation and vasoconstriction. The expression and distribution of KCa subtypes were detected by Western blot and immunohistochemistry. RESULTS: Both the BKCa channel activator NS1619 and the IKCa/SKCa channel activator NS309 evoked significant IMA relaxation. Homocysteine exposure suppressed NS1619-induced relaxation whereas showed no influence on NS309-induced response. Blockade of BKCa but not IKCa and SKCa subtypes significantly suppressed acetylcholine-induced relaxation and enhanced U46619-induced contraction. Homocysteine compromised the vasodilating activity of the BKCa subtype in IMA, associated with a lowered protein level of the BKCa ß1-subunit. Homocysteine potentiated the role of IKCa and SKCa subtypes in mediating endothelium-dependent relaxation without affecting the expression of these channels. CONCLUSIONS: Homocysteine reduces the expression of BKCa ß1-subunit and compromises the vasodilating activity of BKCa channels in IMA. Unlike BKCa, IKCa and SKCa subtypes are unessential for IMA vasoregulation, whereas the loss of BKCa functionality in hyperhomocysteinemia enhances the role of IKCa and SKCa subtypes in mediating endothelial dilator function. Targeting BKCa channels may form a strategy to improve the postoperative graft performance in CABG patients with hyperhomocysteinemia who receive IMA grafting.

Calcium-activated potassium channel family in coronary artery bypass grafts.

OBJECTIVES: We examined the expression, distribution, and contribution to vasodilatation of the calcium-activated potassium (KCa) channel family in the commonly used coronary artery bypass graft internal thoracic artery (ITA) and saphenous vein (SV) to understand the role of large conductance KCa (BKCa), intermediate-conductance KCa (IKCa), and small-conductance KCa (SKCa) channel subtypes in graft dilating properties determined by endothelium-smooth muscle interaction that is essential to the postoperative performance of the graft. METHODS: Real-time polymerase chain reaction and western blot were employed to detect the messenger RNA and protein level of KCa channel subtypes. Distribution of KCa channel subtypes was examined by immunohistochemistry. KCa subtype-mediated vasorelaxation was studied using wire myography. RESULTS: Both ITA and SV express all KCa channel subtypes with each subtype distributed in both endothelium and smooth muscle. ITA and SV do not differ in the overall expression level of each KCa channel subtype, corresponding to comparable relaxant responses to respective subtype activators. In ITA, BKCa is more abundantly expressed in smooth muscle than in endothelium, whereas SKCa exhibits more abundance in the endothelium. In comparison, SV shows even distribution of KCa channel subtypes in the 2 layers. The BKCa subtype in the KCa family plays a significant role in vasodilatation of ITA, whereas its contribution in SV is quite limited. CONCLUSIONS: KCa family is abundantly expressed in ITA and SV. There are differences between these 2 grafts in the abundance of KCa channel subtypes in the endothelium and the smooth muscle. The significance of the BKCa subtype in vasodilatation of ITA may suggest the potential of development of BKCa modulators for the prevention and treatment of ITA spasm during/after coronary artery bypass graft surgery.

Hydrogen sulfide improves endothelial dysfunction in hypertension by activating peroxisome proliferator-activated receptor delta/endothelial nitric oxide synthase signaling.

OBJECTIVE: We aimed to elucidate the ameliorative effect of hydrogen sulfide (H2S) on endothelium-dependent relaxation disturbances via peroxisome proliferator-activated receptor delta/endothelial nitric oxide synthase (PPARδ/eNOS) pathway activation in hypertensive patients and rats. METHODS: Renal arteries were collected from normotensive and hypertensive patients who underwent nephron-sparing surgery. Renal arteries from 37 patients were cultured with or without sodium H2S (NaHS) 50 µmol/l. The rats were randomly divided into four groups: Sham; Sham + NaHS, two kidneys; one clipped (2K1C); and 2K1C + NaHS. Mean arterial pressure was measured by tail-cuff plethysmography. A microvessel recording technique was used to observe the effect of NaHS on endothelium-dependent relaxation. Plasma H2S concentrations were detected using the monobromobimane method. Real-time PCR and western blotting were used to assess mRNA and protein levels of AT1, cystathionine γ-lyase, PPARδ, and phosphor-eNOS. Laser confocal scanning microscopy measured intracellular NO production in human umbilical vein endothelial cells. RESULTS: NaHS improved endothelial function in hypertensive humans and rats. The 20-week administration of NaHS to 2K1C rats lowered the mean arterial pressure. In human umbilical vein endothelial cells, NaHS improved the AngII-induced production of NO. NaHS upregulated PPARδ expression, increased protein kinase B (Akt) or adenosine monophosphate kinase-activated protein kinase (AMPK) phosphorylation, and enhanced eNOS phosphorylation. A PPARδ agonist could mimic the ameliorative effect of NaHS that was suppressed by PPARδ, AMPK, or Akt inhibition. CONCLUSION: H2S plays a protective function in renal arterial endothelium in hypertension by activating the PPARδ/PI3K/Akt/eNOS or PPARδ/AMPK/eNOS pathway. H2S may serve as an effective strategy against hypertension.

Hydrogen Sulfide Improves Vascular Calcification in Rats by Inhibiting Endoplasmic Reticulum Stress.

In this study, the vitamin D3 plus nicotine (VDN) model of rats was used to prove that H2S alleviates vascular calcification (VC) and phenotype transformation of vascular smooth muscle cells (VSMC). Besides, H2S can also inhibit endoplasmic reticulum stress (ERS) of calcified aortic tissues. The effect of H2S on alleviating VC and phenotype transformation of VSMC can be blocked by TM, while PBA also alleviated VC and phenotype transformation of VSMC that was similar to the effect of H2S. These results suggest that H2S may alleviate rat aorta VC by inhibiting ERS, providing new target and perspective for prevention and treatment of VC.

Cystathionine-ß-Synthase Gene Transfer Into Rostral Ventrolateral Medulla Exacerbates Hypertension via Nitric Oxide in Spontaneously Hypertensive Rats.

BACKGROUND: Rostral ventrolateral medulla (RVLM) plays a crucial role in the central regulation of cardiovascular functions. Cystathionine-ß-synthase (CBS) is a major hydrogen sulfide (H2S)-generating enzyme that has been identified mainly in the brain. The present study was designed to examine CBS expression and determine its roles and mechanisms of regulating sympathetic outflow and blood pressure (BP) in the RVLM in spontaneously hypertensive rats (SHR). METHODS AND RESULTS: CBS expression was decreased in the RVLM in SHR compared to Wistar-Kyoto (WKY) rats. Accumulating evidences suggest that H2S interacts with nitric oxide (NO) to regulate cardiovascular function. Therefore, we hypothesize that the decrease in CBS expression in the RVLM may be involved in the disorder of l-arginine/NO pathway, which subsequently affects BP in SHR. Overexpression of CBS in the RVLM caused significant increases in BP, heart rate, and urinary norepinephrine excretion in SHR but not in WKY. Acute experiments were carried out at day 7 after gene transfer. NO metabolite levels, neuronal NO synthase, and γ-amino butyric acid were decreased in SHR after CBS gene transfer. Furthermore, pressor responses to microinjection of NG-monomethyl-l-arginine into RVLM were blunt in SHR transfected with AdCBS compared to SHR transfected with AdEGFP. CONCLUSIONS: Overexpression of CBS in the RVLM elicits enhanced pressor responses in SHR, but not in WKY, and the NO system is involved in these effects. The results suggest that alterations of H2S signaling in the brain may be associated with the development of hypertension.

Gene transfer of cystathionine ß-synthase into RVLM increases hydrogen sulfide-mediated suppression of sympathetic outflow via KATP channel in normotensive rats.

Hydrogen sulfide has been shown to have a sympathoinhibitory effect in the rostral ventrolateral medulla (RVLM). The present study examined the function of cystathionine ß-synthase (CBS)/hydrogen sulfide system in the RVLM, which plays a crucial role in the control of blood pressure and sympathetic nerve activity. Adenovirus vectors encoding CBS (AdCBS) or enhanced green fluorescent protein (AdEGFP) were transfected into the RVLM in normotensive rats. Identical microinjection of AdCBS into the RVLM had no effect on systolic blood pressure and heart rate (HR) in conscious rats. Acute experiments were performed at day 7 after gene transfer in anesthetized rats. Microinjection of the CBS inhibitors hydroxylamine (HA) or amino-oxyacetate into the RVLM produced an increase in the renal sympathetic nerve activity (RSNA), mean arterial pressure (MAP), and HR. There was a potentiation of the increases in RSNA, MAP, and HR because of the CBS inhibitors in AdCBS-injected rats compared with AdEGFP-injected rats. Pretreatment with pinacidil, a ATP-sensitive potassium (KATP) channel activator, abolished the effects of HA in two groups. Microinjection of glibenclamide, a KATP channel blocker, produced increases in RSNA, MAP, and HR in AdCBS-injected rats. No changes in behavior were observed in AdEGFP-injected rats. Furthermore, Western blot analysis indicated an increase in the expression of sulfonylurea receptor 2 and inward rectifier K(+) 6.1 in AdCBS-injected rats. These results suggest that the increase in KATP channels in the RVLM may be responsible for the greater sympathetic outflow and pressor effect of HA in AdCBS-injected rats compared with AdEGFP-injected rats.

Mechanistic studies of AVE3085 against homocysteine in endothelial protection.

PURPOSE: Homocysteine (Hcy) is an independent risk factor for cardiovascular diseases that impairs endothelial function. We investigated whether the impaired endothelial function can be restored by the eNOS transcription enhancer AVE3085 in porcine coronary arteries. The effects of AVE3085 against Hcy on eNOS-NO function were studied and further investigations were conducted to reveal the role of arginase and the signaling pathway of eNOS activation in the effect of AVE3085 on endothelial dysfunction caused by Hcy. METHODS: Myograph study of vasorelaxation, electrochemical measurement of NO, RT-PCR and Western blot analysis of eNOS, iNOS expression, and eNOS phosphorylation were performed. Arginase activity was determined by urea production and O2 (.-) generation by lucigenin-enhanced chemiluminenscence. RESULTS: Exposure to Hcy for 24 h attenuated bradykinin-induced relaxation and NO release, downregulated eNOS mRNA expression and protein expressions of eNOS and p-eNOS(Ser1177) whereas it upregulated iNOS expression. AVE3085 restored NO release and relaxation, enhanced eNOS but decreased iNOS expression. Inhibition of protein kinase Akt or PI3 kinase attenuated the effect of AVE3085 on relaxation and eNOS phosphorylation. Arginase activity and O2 (.-) production were inhibited by AVE3085 in Hcy-exposed vessels. CONCLUSIONS: AVE3085 prevents Hcy-induced endothelial dysfunction in coronary arteries by preservation of NO production and suppression of O2 (.-) generation. Preservation of NO is attributed to upregulation of eNOS expression, activation of eNOS via phosphorylation of Ser1177 through a PI3 kinase/Akt-dependent pathway, and inhibition of arginase. Reduction of O2 (.-) generation results from reversal of eNOS uncoupling and inhibition of arginase and iNOS.

Acute hemodynamic responses to adenosine and iloprost in patients with congenital heart defects and severe pulmonary arterial hypertension.

BACKGROUND: Detection of pulmonary vasoreactivity is important for the evaluation of patient with pulmonary arterial hypertension (PAH). The present study aimed to investigate the acute hemodynamic responses to adenosine and iloprost in patients with congenital heart defects (CHDs) and severe PAH. PATIENTS AND METHODS: From Mar 2007 to Nov 2009, 75 patients with severe PAH secondary to left-to-right shunt CHDs underwent acute vasodilator test using aerosolized iloprost (n = 50) or intravenous adenosine (n = 25). The hemodynamics were detected and analyzed. RESULTS: Decreased mean pulmonary arterial pressure (PAP) and pulmonary vascular resistance (PVR) were observed in 39 and 43 patients in the iloprost group, and in 16 and 19 patients in the adenosine group, respectively. However, the mean PAP was higher than 40 mm Hg in both groups. No significant difference was observed in the age and baseline hemodynamics between the patients with the decrease of PVR and mean pulmonary-to-aortic pressure (Pp/Ps) ratio greater than 10% and the remaining patients. Adenosine decreased both PAP and systemic arterial pressure significantly, while iloprost inhalation selectively reduced the PAP and increased the oxygen saturation of femoral arterial blood and the pulmonary-to-systemic flow (Qp/Qs) ratio. Compared with adenosine, iloprost caused a more profound decline in the Pp/Ps ratio, PVR and pulmonary-to-systemic vascular resistance ratio, and increase in the Qp/Qs ratio. CONCLUSIONS: The acute haemodynamic responses to adenosine and iloprost varied among the patients with CHDs and severe PAH. Different to adenosine, inhaled iloprost exerted selective pulmonary vasodilative effects and was beneficial for pulmonary gas exchange.

[Effects of hydrogen sulfide on ouabain-induced delayed after-depolarization and triggered activity in guinea pig papillary muscles].

OBJECTIVE: To explore the effects of hydrogen sulfide (H(2)S) on delayed after-depolarization (DAD) and triggered activity induced by ouabain in male guinea pig papillary muscles and to elucidate the underlying mechanisms. METHODS: An intracellular microelectrode was used to record the patterns of DAD and triggered activity by K-H solution containing ouabain and a high concentration of calcium ion. The latent period, amplitude, duration of DAD and incidence of triggered activity were observed under a pre-treatment with different concentrations of NaHS (donor of H(2)S). The effects of glibenclamide, Bay K8644 and NG-nitro-L-arginine methyl ester (L-NAME) pretreatment on the actions of H(2)S were also studied. RESULTS: NaHS (100, 200 µmol/L) prolonged the latent period of DAD from (12.0 ± 1.0) min to (19.9 ± 1.6) min (P < 0.05), (23.7 ± 1.3) min (P < 0.01), decreased the altitude of DAD from (11.47 ± 0.74) mV to (6.47 ± 0.33) mV, (5.65 ± 0.26) mV (both P < 0.01), shortened the duration of DAD from (205 ± 11) ms to (173 ± 10) ms and (134 ± 7) ms (both P < 0.05). The occurrence of triggered activity was inhibited from 5 samples to 4, 2 and 1 sample in 6 samples. A pretreatment of adenosine triphosphate (ATP)-sensitive potassium channel (K(ATP)) blocker glibenclamide partially blocked the preventive effects of H(2)S on ouabain-induced DAD and triggered activity. The effects of H(2)S were completely blocked by L-type calcium channel agonist Bay K8644 (0.25 µmol/L). However a pretreatment of L-NAME (1 mmol/L), a nitric oxide (NO) synthase inhibitor, showed no effects on H(2)S. CONCLUSION: H(2)S inhibits the ouabain-induced DAD and triggered activity in guinea pig papillary muscles. The opening of K(ATP) channel with a reduced influx of calcium ion may be involved in the protective effects of H(2)S.

New strategy of endothelial protection in cardiac surgery: use of enhancer of endothelial nitric oxide synthase.

BACKGROUND: Endothelial dysfunction related to the loss of nitric oxide (NO) production remains an important issue in cardiac surgery. We examined the hypothesis that AVE3085, a novel compound that enhances eNOS transcription, may protect coronary endothelium against hypoxia-reoxygenation (H-R) injury during cardioplegic arrest and the possible mechanism by which this occurs. METHODS: Porcine coronary small arteries (600-800-microm diameter) were subjected to hypoxia (PO(2) <5 mmHg) in St. Thomas cardioplegic (ST) solution with or without AVE3085 (10 microM) or L-arginine (10 mM) at either 37 or 4 degrees C for 60 min, followed by 30-min reoxygenation. Bradykinin (-10 to -6.5 LogM)-induced, endothelium-dependent relaxation was studied in a myograph in U(46619) precontraction before and after H-R. Protein expressions of eNOS and phosphorylated eNOS at Ser-1177 (p-eNOS(Ser1177)) were also determined. RESULTS: Exposure to ST solution with H-R at both 37 and 4 degrees C markedly reduced bradykinin-induced relaxation in coronary small arteries. Addition of AVE3085 in ST solution at 37 degrees C preserved the vasorelaxant response to bradykinin (95.7 +/- 2.1% vs. 69.2 +/- 6.6%, p < 0.01), with the protective effect comparable to that of L-arginine (96.1 +/- 3.3% vs. 70.6 +/- 8.7%, p < 0.05). eNOS and p-eNOS(Ser1177) expressions in coronary endothelial cells were significantly increased by the addition of AVE3085 in ST solution during hypoxia (p < 0.05). Protection of endothelium-dependent relaxation from H-R by AVE3085 (70.3 +/- 7.2% vs. 90.5 +/- 2.4%, p < 0.05) also reached a level similar to that by L-arginine (69.9 +/- 9.0% vs. 94.7 +/- 3.9%, p < 0.05) at 4 degrees C. CONCLUSIONS: We have demonstrated a new mechanism to protect coronary endothelium from H-R injury by using eNOS enhancers. This may form a new strategy in the future development of cardioplegic/preservation solutions with direct targeting of eNOS expression in coronary vasculature.