Inhibition of Taurine-upregulated Gene 1 Upregulates MiR-34a-5p to Protect against Myocardial Ischemia/Reperfusion via Autophagy Regulation.

BACKGROUND: Taurine upregulated gene 1 (TUG1) has been identified on long noncoding RNA (lncRNA); however, its function in myocardial cells following ischemia/ reperfusion (I/R) injury has not been explored. This study aimed to investigate the role of LncTUG1 in I/R injury by focusing on its relationship with autophagy induction by regulating miR-34a-5p expression. METHODS: We established a myocardial I/R model and H9C2 hypoxia-ischemic and reoxygenation (HI/R) conditions to induce I/R injury. TTC, Western blot, CCK-8 assay, quantitative reverse transcription PCR, flow cytometry, and confocal microscopy were used to assess the size of myocardial infarct, level of some apoptotic-related and autophagy-associated proteins, cell viability, the level of LncRNA TUG1, apoptosis, and autophagy, respectively. RESULTS: The results revealed that a TUG1 knockdown protected against I/R-induced myocardial injury by decreasing the impairment in cardiac function. LncRNA TUG1 expression was increased in a myocardial I/R model and HI/R in H9C2 cells. Moreover, inhibition of LncTUG1 enhanced H9C2 cell viability and protected the cells from HI/R-induced apoptosis. Silencing LncRNA TUG1 promoted HI/R-induced autophagy. Furthermore, TUG1 siRNA upregulated the level of miR-34a-5p compared to the HI/R group. The protective effect of LncRNA TUG1 inhibition on H9C2 cells following HI/R was eliminated by blocking autophagy with an miR-34a-5p inhibitor. CONCLUSION: These findings indicated that inhibiting TUG1 may reduce the extent of myocardial I/R injury by regulating miR-34a-5p. Taken together, these results suggest that LncRNA TUG1 may represent a novel therapeutic target for myocardial I/R injury.

Anomalous left anterior descending coronary artery arising from the right coronary artery: a case report.

Congenital anomalous coronary artery origins are rare, with a prevalence of 0.24-1.6%. Common configurations include left circumflex arising from the right coronary (RCA), left main coronary arising from the right coronary sinus and independent ostia for the left anterior descending (LAD) and left circumflex. One rare configuration that is not well described is the LAD arising from the RCA. A 68-year-old patient presented with non-ST elevation myocardial infarction, underwent coronary artery bypass grafting and was found to have a patent LAD arising from the RCA. In patients presenting with coronary artery disease and anomalous coronary anatomy, careful preoperative planning and intraoperative attention to patient anatomy is essential in identifying high-risk coronary anomalies and providing the correct and optimal treatments for these patients.

Abnormal origins of the heart arteries are rare, affecting an estimated 0.24­1.6% of the general population. The two main coronary arteries are the right coronary artery (RCA) and left main coronary artery, which branches into the left anterior descending (LAD) and left circumflex artery. While several configurations have been described, one exceedingly rare configuration is the LAD arising from the RCA. A 68-year-old patient presenting with an acute coronary event undergoing coronary artery bypass grafting was found to have an LAD arising from the RCA. Patients presenting with coronary artery disease and abnormal coronary anatomy require careful preoperative planning and intraoperative attention to patient anatomy to identify high-risk coronary abnormalities and providing the optimal treatments for these patients.

False lumen patency status and outcomes after endovascular repair of uncomplicated chronic type B dissection.

BACKGROUND: Thoracic endovascular aortic repair (TEVAR) remains a controversial treatment for uncomplicated chronic type B aortic dissection (cTBAD). This study was performed to investigate the postoperative outcomes of TEVAR, such as survival and reintervention, and the risk factors for prognoses. METHODS: In total, 41 patients with uncomplicated cTBAD who underwent TEVAR from 2014 to 2021 were reviewed. The patients were divided into two groups: those with false lumen complete thrombosis (FLCT) and false lumen partial thrombosis (FLPT) based on computed tomography angiography (CTA) images. Kaplan-Meier analysis was performed to estimate survival and freedom from reintervention. Binary logistic analysis was performed to estimate risk factors for partial thrombosis. RESULTS: During a mean follow-up of 31 (1-78) months, five deaths and six reinterventions had occurred at 5 years. By 1 week, thoracic FLCT had occurred in 23 (56.1%) patients and thoracic FLPT had occurred in 18 (43.9%). The rate of freedom from reintervention was significantly lower in the FLCT than in the FLPT group (p = 0.04). The 5-year survival rate of the two groups was not statistically significant (p = 0.14). Risk factors for thoracic FLPT were the distance between the re-entry site and the graft (p = 0.02) and the proximal oversizing ratio (p = 0.04). CONCLUSIONS: TEVAR is an effective and safe treatment for uncomplicated cTBAD and has a low mortality rate. Thoracic FLCT is associated with less reintervention, but overall survival is not impacted by this difference. Patients treated with TEVAR without certain risk factors can have a good prognosis.

[Resveratrol pretreatment improves mitochondrial function and alleviates myocardial ischemia-reperfusion injury by up-regulating mi R-20b-5p to inhibit STIM2].

This study aimed to explore the mechanism of resveratrol(RES) pretreatment in improving mitochondrial function and alleviating myocardial ischemia-reperfusion(IR) injury by inhibiting stromal interaction molecule 2(STIM2) through microRNA-20 b-5 p(miR-20 b-5 p). Ninety rats were randomly assigned into sham group, IR group, IR+RES(50 mg·kg~(-1) RES) group, IR+RES+antagomir NC(50 mg·kg~(-1) RES+80 mg·kg~(-1) antagomir NC) group, and IR+RES+miR-20 b-5 p antagomir(50 mg·kg~(-1) RES+80 mg·kg~(-1) miR-20 b-5 p antagomir) group, with 18 rats/group. The IR rat model was established by ligation of the left anterior descending coronary artery. Two weeks before the operation, rats in the IR+RES group were intraperitoneally injected with 50 mg·kg~(-1) RES, and those in the sham and IR groups were injected with the same dose of normal saline, once a day. Ultrasonic instrument was used to detect the left ventricular internal diameter at end-diastole(LVIDd) and left ventricular internal diameter at end-systole(LVIDs) of rats in each group. The 2,3,5-triphenyte-trazoliumchloride(TTC) method and hematoxylin-eosin(HE) staining were employed to detect the myocardial infarction area and histopathology, respectively. Real-time quantitative PCR(qRT-PCR) was carried out to detect the expression of miR-20 b-5 p in myocardial tissue. Oxygen glucose deprivation/reoxygenation(OGD/R) was performed to establish an OGD/R model of H9 c2 cardiomyocytes. CCK-8 assay was employed to detect H9 c2 cell viability. H9 c2 cells were assigned into the control group, OGD/R group, OGD/R+RES group(25 µmol·L~(-1)), OGD/R+RES+inhibitor NC group, OGD/R+RES+miR-20 b-5 p inhibitor group, mimic NC group, miR-20 b-5 p mimic group, inhibitor NC group, and miR-20 b-5 p inhibitor group. Flow cytometry was employed to detect cell apoptosis. Western blot was employed to detect the expression of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cleaved-cysteine proteinase 3(cleaved-caspase-3), and STIM2 in cells. The mitochondrial membrane potential(MMP) assay kit, reactive oxygen species(ROS) assay kit, and adenosine triphosphate(ATP) assay kit were used to detect the MMP, ROS, and ATP levels, respectively. Dual luciferase reporter gene assay was adopted to verify the targeting relationship between miR-20 b-5 p and STIM2. Compared with the sham group, the modeling of IR increased the myocardial infarction area, LVIDd, LVIDs, and myocardial pathology and down-regulated the expression of miR-20 b-5 p(P<0.05). These changes were alleviated in the IR+RES group(P<0.05). The IR+RES+miR-20 b-5 p antagomir group had higher myocardial infarction area, LVIDd, LVIDs, and myocardial pathology and lower expression of miR-20 b-5 p than the IR+RES group(P<0.05). The OGD/R group had lower viability of H9 c2 cells than the control group(P<0.05) and the OGD/R+RES groups(25, 50, and 100 µmol·L~(-1))(P<0.05). Additionally, the OGD/R group had higher H9 c2 cell apoptosis rate, protein levels of Bax and cleaved caspase-3, and ROS level and lower Bcl-2 protein, MMP, and ATP levels than the control group(P<0.05) and the OGD/R+RES group(P<0.05). The OGD/R+RES+miR-20 b-5 p inhibitor group had higher H9 c2 cell apoptosis rate, protein levels of Bax and cleaved-caspase 3, and ROS level and lower Bcl-2 protein, MMP, and ATP levels than the OGD/R+RES group(P<0.05). miR-20 b-5 p had a targeting relationship with STIM2. The expression of STIM2 was lower in the miR-20 b-5 p mimic group than in the mimic NC group(P<0.05) and lower in the inhibitor NC group than in the miR-20 b-5 p inhibitor group(P<0.05). RES pretreatment can inhibit the expression of STIM2 by promoting the expression of miR-20 b-5 p, thereby improving the function of mitochondria and alleviating myocardial IR damage.

The effect of in situ laser fenestration for total endovascular arch repair in redo aortic dissection.

OBJECTIVE: Treatment of aortic arch pathologies in redo cases is technically challenging. In this study, we assessed early and mid-term outcomes of total endovascular arch repair combined with a new method of in situ laser fenestration. METHODS: Between January 2018 and March 2019, five patients with a history of cardiovascular surgery underwent in situ laser fenestration procedures using the "squid capture technique" for aortic arch pathologies with dissection. All patients were followed up regularly and imaging examinations were performed. The technical success, procedural complications, as well as the early and mid-term mortality and morbidity rates were evaluated. RESULTS: All patients survived the operation and fenestration was technically successful in all of the patients. There was no in-hospital mortality. No patients developed major complications, such as peri-operative strokes, transient ischemic attacks, or spinal cord ischemia. The 11-22 months follow-up (mean, 17 months) was completed by all patients. No endoleaks were discovered; false lumen thromboses and subsequent positive remodeling of the aorta were demonstrated and all in situ laser-fenestrated arteries were patent. CONCLUSIONS: In situ laser fenestration combined with "squid capture technique" was shown to may be an effective and safe option for reconstruction of aortic arch during thoracic endovascular aortic repair. In situ laser fenestration combined with "squid capture technology" was shown to be an effective treatment option for patients with prior history of cardiovascular surgery and who are at high risk for redo open operations.

Severe hemolytic anemia and acute renal failure after mitral valve repair associated with non-endothelialization of artificial chordae tendinae: case report.

BACKGROUND: Mechanical hemolytic anemia and acute renal failure are rare complications of mitral valve repair. CASE PRESENTATION: We report a unique case of severe hemolytic anemia and severe acute renal failure after mitral valve repair using artificial chordae tendinae. Conservative therapy including plasmapheresis and blood transfusion was not effective. The major cause of the mechanical hemolysis was mild mitral regurgitation originating from the centre of the valve and striking the annuloplasty ring. The hemolytic anemia resolved gradually after the replacement of mitral valve. The new artificial chordae tendinae was found to be completely non-endothelialized in the surgery. Non-endothelialization of artificial chordae tendinae may also play a role in the genesis of mechanical anemia. CONCLUSIONS: The major cause of the mechanical hemolysis was mild mitral regurgitation originating from the centre of the valve and striking the annuloplasty ring. Non-endothelialization of foreign materials might be another mechanism of hemolysis after mitral repair.

Parachute mitral valve associated with reticular chordae tendineae in an adult: case report.

BACKGROUND: Parachute mitral valve with reticular chordae tendineae is an extremely rare anomaly. CASE PRESENTATION: We present a case of parachute mitral valve associated with distinctive reticular chordae tendineae in an adult. It was diagnosed from the echocardiogram. The patient was referred for surgery. Valve analysis showed thickened mitral valve leaflets and commissures. The chordae tendinae were lengthy and thick. All the chordae tendinae merged into a solitary papillary muscle. A distinctive reticular fibrous tissue was found on mitral valve apparatus as the chordae tendinae intermixed each other. The only functional communication between the left atrium and the left ventricle was through the reticular spaces. This anomaly was considered to be unrepairable and was replaced with a mechanical valve. CONCLUSIONS: An extremely rare and unique case of parachute mitral valve associated with reticular chordae tendineae was reported. Mitral valve replacement is a reasonable choice in patients with parachute mitral valve with reticular chordae tendineae.

Conservative treatment of left ventricular pseudoaneurysm after mitral valve replacement due to early left ventricular rupture: a case report.

BACKGROUND: Left ventricular pseudoaneurysm due to early left ventricle rupture is a serious complication after cardiac surgery. Urgent surgery is recommended in most cases with a high mortality rate. Conservative treatment of a left ventricular pseudoaneurysm due to early left ventricle rupture is very rare. CASE PRESENTATION: We present a 61-year-old woman with left ventricular pseudoaneurysm after mitral valve replacement due to early left ventricle rupture. This patient was treated in a conservative approach. This patient had an uneventful recovery. She was in good condition and remained asymptomatic 3.5 years after mitral valve surgery. CONCLUSION: This case suggests that medical treatment left ventricular pseudoaneurysm patients has a limited but acceptable role in selected and unusual circumstances.

Spindle cell sarcoma of the right atrium causing right atrial pseudoaneurysm: a case report and review of the literature.

BACKGROUND: Spindle cell sarcomas of the right atrium are extremely rare primary cardiac tumours, with very few cases reported in the medical literature. Pseudoaneurysms caused by cardiac spindle cell sarcoma have never been reported worldwide. CASE PRESENTATION: A 32-year-old woman was referred to our hospital for recurrent pericardial haemorrhagic effusion and pleural effusion. Three-dimensional transthoracic echocardiogram, contrast chest CT, and contrast MRI revealed a pseudoaneurysm on the right side of the right atrium with a thrombus. There was a defect between the pseudoaneurysm and the right atrium. PET-CT suggested that FDG metabolism inhomogeneity increased in the mass in the right atrium. Exfoliative cytology detection of massive pericardial effusion and pleural effusion revealed no tumour cells. Spindle cell sarcoma of the right atrium was not confirmed until the patient underwent right thoracic exploration and biopsy. Before a confirmed diagnosis, symptomatic treatment, such as chest effusion and pericardium effusion drainage, and transfusion of red blood cells were mainly used to relieve the patient's symptoms. Unfortunately, the patient was lost to optimal treatments and passed away 20 days after the pathological diagnosis was made. CONCLUSION: The prognosis of spindle cell sarcomas remains poor due to delays in diagnosis, early metastasis and few available therapeutic options. Recurrent pericardial effusion and pleural effusion, especially in the nature of haemorrhagic effusion, and/or right atrial pseudoaneurysm shown on the transthoracic echocardiogram must be considered and highly suspected as malignancy by patients and physicians. If the diagnosis cannot be confirmed, histopathology should be performed as soon as possible to avoid losing the best treatment opportunity.

In situ laser fenestration for revascularization of aortic arch during treatment for iatrogenic type A aortic dissection.

BACKGROUND: Iatrogenic aortic dissection is a rare and fatal complication. Its treatment was challenging and controversial especially in patients with previous cardiac procedure. This study aimed to present the case of a patient with aortic dissection after previous open cardiac surgery who was successfully treated by in situ laser fenestration for revascularization of aortic arch. CASE PRESENTATION: A 65-year-old man suffered severe aortic and mitral valve regurgitation was treated by open cardiac aortic valve replacement (biological valve, Edwards) and mitral valve repair. During the sixth-month follow-up, computed tomography angiography (CTA) scan revealed an aortic dissection that extended from the ascending aorta to both femoral arteries. After stabilized by medical treatment, the patient was treated by endovascular stent-graft implantation and in situ laser (holmium laser, energy: 0 5 J, frequency: 5 Hz.) fenestration for revascularization of aortic arch in our one-stop hybrid operating room. The patient recovered without any clinical complication and was discharged 5 days after the procedure. CONCLUSIONS: Our work suggested that in situ laser fenestration for revascularization of aortic arch is a feasible, effective, and safe treatment in patients with iatrogenic aortic dissection.

Follistatin-Like 1 Attenuates Ischemia/Reperfusion Injury in Cardiomyocytes via Regulation of Autophagy.

BACKGROUND: The cardioprotective effect of FSTL1 has been extensively studied in recent years, but its role in myocardial ischemia/reperfusion injury (IRI) is unclear. In this study, we investigated the effect of FSTL1 pretreatment on myocardial IRI as well as the possible involvement of autophagic pathways in its effects. METHODS: The effects of FSTL1 on the viability and apoptosis of rat cardiomyocytes were investigated after exposure of cardiomyocytes to hypoxia/ischemia by using the CCK-8 assay and Annexin V/PI staining. Further, western blot analysis was used to detect the effects of FSTL1 pretreatment on autophagy-associated proteins, and confocal microscopy was used to observe autophagic flux. To confirm the role of autophagy, the cells were treated with the autophagy promoter rapamycin or the autophagy inhibitor 3-methyladenine, and cell viability and apoptosis during IRI were observed. These effects were also observed after treatment with rapamycin or 3-methyladenine followed by FSTL1 administration and IRI. RESULTS: FSTL1 pretreatment significantly increased viability and reduced apoptosis in cardiomyocytes exposed to hypoxia/ischemia conditions. Further, FSTL1 pretreatment affected the levels of the autophagy-related proteins and enhanced autophagic flux during IRI. In addition, cell viability was enhanced and apoptosis was decreased by rapamycin treatment, while these effects were reversed by 3-MA treatment. However, when the myocardial cells were pretreated with rapamycin or 3-methyladenine, there was no significant change in their viability or apoptosis with FSTL1 treatment during IRI. CONCLUSIONS: FSTL1 plays a protective role in myocardial IRI by regulating autophagy.

Spermine ameliorates ischemia/reperfusion injury in cardiomyocytes via regulation of autophagy.

Myocardial infarction could result in high morbidity and mortality and heart diseases of children have becoming prevalent. Functions of spermine administration on cardiomyocytes remain unknown. The present study was designed to investigate the role of spermine pretreatment on myocardial ischemia/reperfusion injury (IRI). A cell model of simulated ischemia/reperfusion injury was established by incubating neonatal Sprague-Dawley rat cardiomyocytes in ischemia medium and re-cultured in normal medium. Of note, spermine pretreatment significantly reduced apoptosis and increased viability of immature cardiomyocytes. Spermine pretreatment enhanced autophagic flux as determined by confocal microscopy and transmission electron microscopy. Furthermore, proteins of mammalian target of rapamycin (mTOR) pathway were significantly reduced in response to spermine pretreatment during IRI, while proteins related to autophagy were up-regulated. The cell viability was enhanced and apoptosis decreased by rapamycin after spermine pretreatment, while these were reversed by 3-methyladenine. However, when immature cardiomyocytes were pretreated with rapamycin or 3-methyladenine, followed by IRI and spermine administration, no significant changes of viability and apoptosis were observed. In conclusion, this study suggests that spermine is a potential novel approach for preventing IRI, especially in children.

Mitral valve repair for double-orifice mitral valve.

We present an eight year-old girl who required an operation for moderate mitral insufficiency associated with partial atrioventricular septal defect. Echocardiography disclosed an ostium primum atrial septal defect and double-orifice mitral valve with moderate mitral regurgitation secondary to a cleft in the anterior leaflet and prolapse of the anterior leaflet. Intraoperative inspection revealed that the chordae from each orifice were attached to a single papillary muscle which resulted in a unique double-orifice mitral valve. Mitral valve repair using chordal shortening and cleft closure was successfully performed. Postoperative echocardiography observed trivial MR and no mitral stenosis.

Anterolateral minithoracotomy versus median sternotomy for mitral valve disease: a meta-analysis.

OBJECTIVE: Mitral valve disease tends to be treated with anterolateral minithoracotomy (ALMT) rather than median sternotomy (MS), as ALMT uses progressively smaller incisions to promote better cosmetic outcomes. This meta-analysis quantifies the effects of ALMT on surgical parameters and post-operative outcomes compared with MS. METHODS: One randomized controlled study and four case-control studies, published in English from January 1996 to January 2013, were identified and evaluated. RESULTS: ALMT showed a significantly longer cardiopulmonary bypass time (P=0.001) and aortic cross-clamp time (P=0.05) compared with MS. However, the benefits of ALMT were evident as demonstrated by a shorter length of hospital stay (P<0.00001). According to operative complications, the onset of new arrhythmias following ALMT decreased significantly as compared with MS (P=0.05); however, the incidence of peri-operative mortality (P=0.62), re-operation for bleeding (P=0.37), neurologic events (P=0.77), myocardial infarction (P=0.84), gastrointestinal complications (P=0.89), and renal insufficiency (P=0.67) were similar to these of MS. Long-term follow-up data were also examined, and revealed equivalent survival and freedom from mitral valve events. CONCLUSIONS: Current clinical data suggest that ALMT is a safe and effective alternative to the conventional approach and is associated with better short-term outcomes and a trend towards longer survival.

Cardiac-targeting magnetic lipoplex delivery of SH-IGF1R plasmid attenuate norepinephrine-induced cardiac hypertrophy in murine heart.

Recent studies have demonstrated a number of molecular mechanisms contributing to the initiation of cardiac hypertrophy response to pressure overload. IGF1R (insulin-like growth factor-1 receptor), an important oncogene, is overexpressed in hypertrophic heart and mediates the hypertrophic pathology process. In this study, we applied with liposomal magnetofection that potentiated gene transfection by applying an external magnetic field to enhance its transfection efficiency. Liposomal magnetofection provided high efficiency in transgene expression in vivo. In vivo, IGF1R-specific-shRNA (small-hairpin RNA) by magnetofection inhibited IGF1R protein expression by 72.2 ± 6.8, 80.7 ± 9.6 and 84.5 ± 5.6%, at 24, 48 and 72 h, respectively, after pGFPshIGF1R injection, indicating that liposomal magnetofection is a promising method that allows the targeting of gene therapy for heart failure. Furthermore, we found that the treated animals (liposomal magnetofection with shIGF1R) showed reduced septal and posterior wall thickness, reduced HW:BWs (heart weight-to-body weights) compared with controls. Moreover, we also found that liposomal magnetofection-based shIGF1R transfection decreased the expression level of p-ERK (phosphorylated extracellular-signal-regulated kinase)1/2, p-AKT1 (phosphorylated protein kinase B1) compared with untreated hearts. These results suggested that liposomal magnetofection-mediated IGF1R-specific-shRNA may be a promising method, and suppression the IGF1R expression inhibited norepinephrine-induced cardiac hypertrophic process via inhibiting PI3K (phosphoinositide 3-kinase)/AKT pathway.

MicroRNA-150 aggravates H2O2-induced cardiac myocyte injury by down-regulating c-myb gene.

MicroRNAs (miRNAs) are one class of non-coding RNAs that play an important role in post-transcriptional regulation via the degradation or translational inhibition of their target genes. MicroRNA-150 (miR-150) plays a vital role in regulating the development of B and T lymphocytes. Although the dysregulation of miR-150 was confirmed in human myocardial infarction, little is known regarding the biological functions of miR-150 in response to reactive oxygen species (ROS)-mediated gene regulation in cardiac myocytes. Using quantitative real-time reverse transcription-polymerase chain reaction, we demonstrated that the level of miR-150 was up-regulated in cardiac myocytes after treatment with hydrogen peroxide (H2O2). To identify the potential roles of miR-150 in H2O2-mediated gene regulation, we modulated expression of miR-150 using miR-150 inhibitor and miR-150 mimics. Results showed that silencing expression of miR-150 decreased H2O2-induced cardiac cell death and apoptosis. In lymphocytes, c-myb was a direct target of miR-150. In cardiac myocytes, we found that c-myb was also involved in miR-150-mediated H2O2-induced cardiac cell death. These results suggested that miR-150 participates in H2O2-mediated gene regulation and functional modulation in cardiac myocytes. MiR-150 may play an essential role in heart diseases related to ROS, such as cardiac hypertrophy, heart failure, myocardial infarction, and myocardial ischemia/reperfusion injury.

Tissue distribution and cancer growth inhibition of magnetic lipoplex-delivered type 1 insulin-like growth factor receptor shRNA in nude mice.

The targeted delivery of therapeutic genes into specific tissues, as well as the determination of the biological fate and potential toxicity of nanoparticles, remains a highly relevant challenge for gene-based therapies. Type 1 insulin-like growth factor receptor (IGF-1R), an important oncogene, is frequently over-expressed in lung cancer and mediates cancer cell proliferation as well as tumor growth. In our previous studies, we have successfully applied gene delivery mediated by commercially available nanoparticles (CombiMAG) under a magnetic field, which suppresses IGF-1R expression in a non-small cell lung cancer cell line (A549) in vitro. In the present study, we aimed to investigate the biological distribution and target tumor suppression of magnetofection, as well as its potential toxicity via CombiMAG-carrying plasmids expressing green fluorescent protein (GFP) and short hairpin RNAs (shRNAs) targeting IGF-1R (pGFPshIGF-1Rs) in tumor-bearing mice. The peak expression in various organs appeared 48 h after transfection. Transgene expression via magnetofection was 3-fold improvement than via lipofection. On the 30th day after injection, the tumor size and weight of the CombiMAG-treated group (789.32 ± 39.43 mm(3), 105.5 ± 6.1 mg) were significantly decreased compared with those of the lipofection group (893.83 ± 31.23 mm(3), 164.5 ± 9.1 mg; P< 0.05), and the suppression rate was ∼36%. After a 30-day observation, the injection of CombiMAG did not cause any apparent toxicity. Therefore, IGF-1R shRNA nanoparticles can be valuable and safe delivery agents for RNA interference therapy to tumors in vivo.

Efficient inhibition of human cytomegalovirus UL122 gene expression in cell by small interfering RNAs.

In order to develop a gene therapy to human cytomegalovirus (HCMV), RNA interference (RNAi) was employed to inhibit the expression of HCMV UL122 gene in vitro. Recombinant vector pUL122-EGFP, which expressed UL122-EGFP fusion protein, and recombinant vectors psi122-1, psi122-2 and psi122-3, which expressed small interfering RNAs (siRNAs) targeted to UL122 were contransfected into AD293 cells. The fluorescence signal of pUL122-EGFP was greatly suppressed by psi122-1 and psi122-2, with an inhibitory rate of 82.0% +/- 1.0% and 79.5% +/- 2.5%, respectively. The mRNA of pUL122-EGFP of the cells transfected with psi122-1 and psi122-2 was decreased 97.3% +/- 0.6% and 98.0% +/- 0.1%, respectively. Vector psi122-3 showed a slightly low suppression rate. Therefore, it may be concluded that plasmids encoding siRNAs targeted to UL122 is able to in vitro reduce markedly the expression of UL122-EGFP. And it is very likely that the psi122-1 and psi122-2 are potentially efficacious siRNAs in the gene therapy of HCMV infection in vivo, in which further investigations are required. This study is expected to greatly facilitate the use of the RNAi technology for the anti-HCMV studies.