Phytofabrication of Nanoparticles as Novel Drugs for Anticancer Applications.

Cancer is one of the foremost causes of death globally and also the major stumbling block of increasing life expectancy. Although the primary treatment of surgical resection, chemotherapy, and radiotherapy have greatly reduced the mortality of cancer, the survival rate is still low because of the metastasis of tumor, a range of adverse drug reactions, and drug resistance. For all this, it is relevant to mention that a growing amount of research has shown the anticarcinogenic effect of phytochemicals which can modulate the molecular pathways and cellular events include apoptosis, cell proliferation, migration, and invasion. However, their pharmacological potential is hindered by their low water solubility, low stability, poor absorption, and rapid metabolism. In this scenario, the development of nanotechnology has created novel formulations to maximize the potential use of phytochemicals in anticancer treatment. Nanocarriers can enhance the solubility and stability of phytochemicals, prolong their half-life in blood and even achieve site-targeting delivery. This review summarizes the advances in utilizing nanoparticles in cancer therapy. In particular, we introduce several applications of nanoparticles combined with apigenin, resveratrol, curcumin, epigallocatechin-3-gallate, 6-gingerol, and quercetin in cancer treatment.

Comparison of Intracoronary and Intravenous Ultrasound-targeted Microbubble Destruction-mediated Ang1 Gene Transfection on Left Ventricular Remodeling in Canines With Acute Myocardial Infarction.

Intravenous ultrasound-targeted microbubble destruction (IV-UTMD) has made distinct but limited progress in gene therapy. Intracoronary (IC) injection may lead to more gene transfection than IV injection. This study compared the therapeutic effects of IC-UTMD-mediated and conventional IV-UTMD-mediated gene transfection in acute myocardial infarction (MI). A canine MI model was successfully established through transcatheter coronary artery embolism, and the animals were divided into several treatment groups: IC injection with UTMD and the negative control plasmid (IC-UTMD); IC injection of the angiopoietin 1 (Ang1) plasmid (IC-Ang1); IC injection with UTMD and the Ang1 plasmid (IC-UTMD-Ang1); and IV injection with UTMD and the Ang1 plasmid (IV-UTMD-Ang1). At 12 hours after injection, more green fluorescence was observed from the fluorescein isothiocyanate-labeled Ang1 plasmid in the IC-UTMD-Ang1 group. After 1 month, compared with the IV-UTMD-Ang1 group, echocardiography showed that the IC-UTMD-Ang1 group exhibited increased left ventricular systolic function and myocardial infusion, with lower fibrous tissue levels and higher blood vessel density and Ang1 mRNA and protein levels. Similar cardiac troponin I and N-terminal pro-B type natriuretic peptide levels were observed in all groups. Compared with IV-UTMD, IC-UTMD can enhance Ang1 plasmid transfection efficiency after MI, promote gene expression and angiogenesis, and improve left ventricular remodeling without decreasing safety.

The differences in left atrial function between ischemic and idiopathic dilated cardiomyopathy patients: A two-dimensional speckle tracking imaging study.

PURPOSE: To evaluate left atrial (LA) function in patients with ischemic (ICM) or idiopathic dilated (DCM) cardiomyopathy via two-dimensional speckle-tracking imaging. METHODS: We measured the LA maximum volume, minimum volume, and volume before the atrial systole, and calculated total emptying volume, expansion index, active emptying volume, and fraction. We measured strain and strain rate during systole and late diastole using two-dimensional speckle-tracking imaging, and analyzed correlations between variables. RESULTS: We found no significant differences in LA size, left ventricle (LV) end-diastole diameter, LV ejection fraction (EF), E/A, E/e', deceleration time of the E wave, and effective mitral regurgitant orifice area between the DCM and the ICM group. However, the LA expansion index, active EF, systolic and late diastolic strain, and strain rate were lower in the ICM group (p < 0.05). The expansion index and active EF were positively correlated with the systolic strain rate and the absolute value of the late diastolic strain rate, respectively. CONCLUSIONS: LA basic echocardiographic variables did not reflect the differences between ICM and DCM patients, but the systolic and late diastolic strain, as well as the strain rate, were lower in DCM patients. Two-dimensional speckle-tracking imaging is a promising method to differentiate these patients. © 2016 Wiley Periodicals, Inc. J Clin Ultrasound 44:437-445, 2016.

Progressive decay of Ca2+ homeostasis in the development of diabetic cardiomyopathy.

BACKGROUND: Cardiac dysfunction in diabetic cardiomyopathy may be associated with abnormal Ca2+ homeostasis. This study investigated the effects of alterations in Ca2+ homeostasis and sarcoplasmic reticulum Ca2+-associated proteins on cardiac function in the development of diabetic cardiomyopathy. METHODS: Sprague-Dawley rats were divided into 4 groups (n = 12, each): a control group, and streptozotocin-induced rat models of diabetes groups, examined after 4, 8, or 12 weeks. Evaluations on cardiac structure and function were performed by echocardiography and hemodynamic examinations, respectively. Cardiomyocytes were isolated and spontaneous Ca2+ spark images were formed by introducing fluorescent dye Fluo-4 and obtained with confocal scanning microscopy. Expressions of Ca2+-associated proteins were assessed by Western blotting. RESULTS: Echocardiography and hemodynamic measurements revealed that cardiac dysfunction is associated with the progression of diabetes, which also correlated with a gradual but significant decline in Ca2+ spark frequency (in the 4-, 8- and 12-week diabetic groups). However, Ca2+ spark decay time constants increased significantly, relative to the control group. Expressions of ryanodine receptor 2 (RyR2), sarcoplasmic reticulum Ca2+-2ATPase (SERCA) and Na+/Ca2+ exchanger (NCX1) were decreased, together with quantitative alterations in Ca2+regulatory proteins, FKBP12.6 and phospholamban progressively and respectively in the diabetic rats. CONCLUSIONS: Ca2+ sparks exhibited a time-dependent decay with progression of diabetic cardiomyopathy, which may partly contribute to cardiac dysfunction. This abnormality may be attributable to alterations in the expressions of some Ca2+-associated proteins.

Molecular epidemiological investigation of G6PD deficiency in Yangjiang region, western Guangdong province.

Objectives: The prevalence of G6PD deficiency has not been reported in Yangjiang, a western city in Guangdong province. This study aims to investigate the molecular characteristics of G6PD deficiency in this region. Methods: Blood samples were collected from adults at a local hospital to screen for G6PD deficiency. The deficient samples were subjected to further analysis using PCR and reverse dot blot to determine the specific G6PD variants. Results: Among the 3314 male subjects, 250 cases of G6PD deficiency were found using the G6PD enzyme quantitative assay, resulting in a prevalence of 7.54% (250/3314) in the Yangjiang region. The prevalence of G6PD deficiency in females was 3.42% (176/5145). Out of the 268 cases of G6PD deficiency tested for G6PD mutations, reverse dot blot identified 20 different G6PD variants. The most common G6PD variant was c.1388G>A (81/268), followed by c.1376G>T (48/268), c.95A>G (32/268), c.1024C>T (9/268), c.392G>T (7/268), and c.871G>A/c.1311C>T (6/268). It was observed that c.871G>A was always linked to the polymorphism of c.1311C>T in this population. Conclusion: This investigation into G6PD deficiency in this area is expected to significantly improve our understanding of the prevalence and molecular characterization of this condition.

Evaluation of microwave-assisted extraction for aristolochic acid from Aristolochiae Fructus by chromatographic analysis coupled with nephrotoxicity studies.

In this paper, a microwave-assisted extraction (MAE) method was established for aristolochic acid-I from Aristolochiae Fructus, and the advantage of MAE was evaluated by chromatographic analysis coupled with nephrotoxicity studies. The experimental parameters of MAE for aristolochic acid-I in Aristolochiae Fructus were investigated and MAE was compared with Soxhlet extraction and ultrasound-assisted extraction in terms of extraction yields and extraction conditions. Under the optimum conditions, MAE could provide higher extraction yields of aristolochic acid-I (1.10 mg/g) than ultrasound-assisted extraction (0.82 mg/g) and Soxhlet extraction (0.95 mg/g), in addition to using less solvent and having a shorter extraction time. Furthermore, the nephrotoxicities of the extracts of Aristolochiae Fructus from different extraction procedures were investigated in Sprague-Dawley rats. The results of nephrotoxicity studies of, for example, general conditions, biochemistry parameters and histopathology examination showed no significantly differences in the nephrotoxicity levels of the extracts from MAE and that from Soxhlet extraction. These results indicated that MAE technique is a simple, rapid and effective extraction method, and the microwave irradiation during MAE procedure did not have any influence on the nephrotoxicity of Aristolochiae Fructus compared with Soxhlet extraction.

Misdiagnosis of unroofed coronary sinus syndrome as an ostium primum atrial septal defect by echocardiography: A case report.

BACKGROUND: Unroofed coronary sinus syndrome (UCSS) is a rare congenital heart disease, which has variable morphologic features and is strongly associated with persistent left superior vena cava (PLSVC). However, it is often difficult to visualize the left-to-right shunt pathway through the CS by transthoracic echocardiography (TTE). CASE SUMMARY: A 37-year-old female was admitted to the hepatological surgery department of a hospital with complaint of subxiphoid pain that had started 1 wk prior. Physical examination revealed a grade 3/6 systolic murmur at the left margin of the sternum, between the 2nd and 3rd intercostal cartilage. The patient underwent echocardiography and was diagnosed with ostium primum atrial septal defect (ASD); thus, she was subsequently transferred to the cardiovascular surgery department. A second TTE evaluation before surgery showed type IV UCSS with secundum ASD. Right-heart contrast echocardiography (RHCE) showed that the right atrium and right ventricle were immediately filled with microbubbles, but no microbubble was observed in the CS. Meanwhile, negative filling was observed at the right atrium orifice of the CS and right atrium side of the secundum atrial septal. RHCE identified UCSS combined with secundum ASD but without PLSVC in this patient. CONCLUSION: This rare case of UCSS highlights the value of TTE combined with RHCE in confirming UCSS with ASD or PLSVC.

Pseudomonas aeruginosa-related effusive-constrictive pericarditis diagnosed with echocardiography: A case report.

BACKGROUND: Effusive-constrictive pericarditis (ECP) is an uncommon pericardial syndrome. Careful echocardiographic examination may provide helpful information not only for diagnosing but also for managing ECP. ECP has various etiologies; however, Pseudomonas aeruginosa (P. aeruginosa) infection has not been reported as a cause to date. Herein, we present a rare case of ECP caused by P. aeruginosa infection, which was followed up using echocardiography. CASE SUMMARY: A 30-year-old man was admitted to our hospital with a 2-mo history of cough, dyspnea, bloating, palpitations, and lower-extremity edema. The patient was initially diagnosed with pericardial effusion by transthoracic echocardiography. Drainage of pericardial effusion was performed to relieve the clinical symptoms. A follow-up echocardiogram showed that the pericardial effusion had decreased; however, the right atrial pressure continued to increase, and signs of constrictive pericarditis were observed upon a more comprehensive inspection. Therefore, the diagnosis of ECP was established based on the comprehensive pre- and post-pericardiocentesis echocardiographic findings. An urgent pericardectomy was subsequently performed, which significantly relieved the patient's clinical symptoms, and the signs of pericardial constriction on echocardiography improved. Pericardial effusion and pericardial culture showed growth of P. aeruginosa. CONCLUSION: ECP induced by P. aeruginosa infection remains a rare disease. The presence of echocardiographic features of constrictive pericarditis after pericardiocentesis therapy is highly indicative of ECP.

Ultrasound-targeted cationic microbubbles combined with the NFκB binding motif increase SDF-1α gene transfection: A protective role in hearts after myocardial infarction.

Treatment of myocardial infarction (MI) remains a major challenge. The chemokine family plays an important role in cardiac injury, repair, and remodeling following MI, while stromal cell-derived factor-1 alpha (SDF-1α) is the most promising therapeutic target. This study aimed to increase SDF-1α expression using a novel gene delivery system and further explore its effect on MI treatment. In this study, two kinds of plasmids, human SDF-1α plasmid (phSDF-1α) and human SDF-1α- nuclear factor κB plasmid (phSDF-1α-NFκB), were constructed and loaded onto cationic microbubble carriers, and the plasmids were released into MI rabbits by ultrasound-targeted microbubble destruction. The transfection efficiency of SDF-1α and the degree of heart repair were further explored and compared. In the MI rabbit models, transfection with phSDF-1α-NFκB resulted in higher SDF-1α expression in peri-infarct area compared with transfection with phSDF-1α or no transfection. Upregulation of SDF-1α was shown beneficial to these MI rabbit models, as demonstrated with better recovery of cardiac function, greater perfusion of the myocardium, more neovascularization, smaller infarction size and thicker infarct wall 1 month after treatment. Ultrasound-targeted cationic microbubbles combined with the NFκB binding motif could increase SDF-1α gene transfection, which would play a protective role after MI.

Prognostic value of myocardial contrast echocardiography in acute anterior wall ST-segment elevation myocardial infarction with successful epicardial recanalization.

Although myocardial contrast echocardiography (MCE) can evaluate microvascular perfusion abnormalities, its prognostic value is uncertain in acute anterior wall ST-Segment elevation myocardial infarction (STEMI) with successful epicardial recanalization. Therefore, the study aims to investigate the prognostic role of qualitative and quantitative MCE in acute anterior wall STEMI with successful epicardial recanalization. 153 STEMI patients were assessed by MCE within 7 days after successful epicardial recanalization. Qualitative perfusion parameters (microvascular perfusion score index, MPSI) and quantitative perfusion parameters (A, ß, and Aß) were acquired using a 17-segment model. And corrected A and Aß were calculated. Patients were all followed for major adverse cardiovascular events (MACEs). During median follow-up of 27 (4) months, 39 (25.49%) patients experienced MACEs, while 114 (74.51%) were free from MACEs. Patients with MACEs had higher MPSI (1.65 ± 0.13 vs. No-MACEs 1.35 ± 0.20, P < 0.001), lower ß (1.09 ± 0.19 s-1 vs. No-MACEs 1.34 ± 0.30 s-1, P < 0.001), corrected A (0.17 ± 0.03 dB vs. No-MACEs 0.19 ± 0.04 dB, P = 0.039) and lower corrected Aß (0.19 ± 0.06 dB/s vs. No-MACEs 0.25 ± 0.08 dB/s, P < 0.001). MPSI of 1.44 provided an area under the curve (AUC) of 0.872, while ß of 1.18 s-1 and corrected Aß of 0.22 dB/s provided AUCs of 0.759 and 0.724, respectively. The combination of MPSI, ß and corrected Aß provided an increased AUC of 0.964 (all P < 0.05). Time-dependent ROC analysis showed that the AUCs of the MPSI, ß, corrected Aß and the combination at 1, 1.5 and 2 years indicated a strong predictive power for MACEs (AUC = 0.900/0.894/0.881 for MPSI, 0.648/0.704/0.732 for ß, 0.674/0.686/0.722 for corrected Aß, and 0.947/0.962/0.967 for the combination, respectively). Patients with MPSI < 1.44, ß > 1.18 s-1, or corrected Aß > 0.22 dB/s had lower event rate (all Log Rank P ≤ 0.001). MPSI, ß, corrected Aß, GLS and WBC were independent predictors of MACEs with adjusted hazard ratio of 34.41 (8.18-144.87), P < 0.001 for MPSI; 39.29 (27.46-65.44), P < 0.001 for ß; 8.93 (1.46-54.55), P = 0.018 for corrected Aß; 10.88 (2.83-41.86), P = 0.001 for GLS; and 1.43 (1.16-1.75), P = 0.001 for WBC. Qualitative and quantitative MCE can accurately predict MACEs in acute anterior wall STEMI with successful epicardial recanalization, and their combined predictive value is higher.

L2Δ13, a splicing isoform of lysyl oxidase-like 2, causes adipose tissue loss via the gut microbiota and lipid metabolism.

Obesity is primarily characterized by the dysregulation of lipid metabolism and gut microbiota. Here, we found that the body weight of transgenic mice overexpressing L2Δ13, a selectively spliced isoform of lysyl oxidase-like 2 (LOXL2), was lower than that of wild-type (WT) mice. Numerous microbiotas were significantly changed and most microbial metabolites were abnormal in L2Δ13 mice. Lipid metabolites in feces were negatively correlated with those in plasma, suggesting that L2Δ13 may affect lipid uptake, and potentially, adipose tissue homeostasis. This was supported by the weight loss and decreased area of adipose tissue in L2Δ13 mice. Adipogenic differentiation of primary stromal vascular fraction cells showed that the lipid droplets of L2Δ13 cells were significantly smaller than those of WT cells. Adipocyte differentiation-associated genes were also downregulated in adipose tissue from L2Δ13 mice. Thus, L2Δ13 can induce adipose tissue loss in mice by affecting gut microbiota homeostasis and multi-tissue lipid metabolism.

Right-heart contrast echocardiography reveals missed patent ductus arteriosus in a postpartum woman with pulmonary embolism: A case report.

BACKGROUND: As an established, simple, inexpensive, and surprisingly effective diagnostic tool, right-heart contrast echocardiography (RHCE) might help in solving a vexing diagnostic problem. If performed appropriately and interpreted logically, RHCE allows for differentiation of various usual and unusual right-to-left shunts based on the site of injection and the sequence of microbubble appearance in the heart. CASE SUMMARY: A 31-year-old woman was readmitted to hospital with a 2-mo history of worsening palpitation and chest distress. Two years prior, she had been diagnosed with postpartum pulmonary embolism by conventional echocardiography and computed tomography angiography. While the latter showed no sign of pulmonary artery embolism, the former showed pulmonary artery hypertension, moderate insufficiency, and mild stenosis of the aortic valve. RHCE showed microbubbles appearing in the left ventricle, slightly delayed after right-heart filling with microbubbles; no microbubbles appeared in the left atrium and microbubbles' appearance in the descending aorta occurred nearly simultaneous to right pulmonary artery filling with microbubbles. Conventional echocardiography was re-performed, and an arterial horizontal bidirectional shunt was found according to Doppler enhancement effects caused by microbubbles. The original computed tomography angiography findings were reviewed and found to show a patent ductus arteriosus. CONCLUSION: RHCE shows a special imaging sequence for unexplained pulmonary artery hypertension with aortic valve insufficiency and simultaneous patent ductus arteriosus.

Tuberous sclerosis complex-lymphangioleiomyomatosis involving several visceral organs: A case report.

BACKGROUND: Lymphangioleiomyomatosis (LAM) is a rare cystic lung disease characterized by the proliferation, metastasis, and infiltration of smooth muscle cells in the lung and other tissues, which can be associated with tuberous sclerosis complex (TSC). The disorder of TSC has a variable expression, and there is great phenotypic variability. CASE SUMMARY: A 32-year-old Chinese woman with a history of multiple renal angioleiomyolipoma presented with a productive cough persisting for over 2 wk. High-resolution chest computed tomography revealed interstitial changes, multiple pulmonary bullae, bilateral pulmonary nodules, and multiple fat density areas of the inferior mediastinum. Conventional and contrast ultrasonography revealed multiple high echogenic masses of the liver, kidneys, retroperitoneum, and inferior mediastinum. These masses were diagnosed as angiomyolipomas. Pathology through thoracoscopic lung biopsy confirmed LAM. Furthermore, high-throughput genome sequencing of peripheral blood DNA confirmed the presence of a heterozygous mutation, c.1831C>T (p.Arg611Trp), of the TSC2 gene. The patient was diagnosed with TSC-LAM. CONCLUSION: We highlight a rare case of TSC-LAM and the first report of a mediastinum lymphangioleiomyoma associated with TSC-LAM.

[Effect of a novel injectable hydrogel implant on ventricular remodeling in rabbits with experimental myocardial infarction].

OBJECTIVE: To observe whether injectable hydrogel implantation could prevent adverse cardiac remodeling and preserve cardiac function in rabbits with myocardial infarction (MI). METHODS: A novel injectable hydrogel, the copolymer MPM/alpha-CD, which self-assembled between alpha-cyclodextrin and methoxy polyethylene glycol-poly (caprolactone)-(dodecanedioic acid)-poly (caprolactone)-methoxypolyethylene glycol triblock polymer, was synthesized by chemical crosslinking and characterized by biocompatibility and biodegradability. Experimental MI was induced in male rabbits by coronary artery ligation. The MI rabbits were randomly divided into hydrogel group (200 microl MPM/alpha-CD were injected into the infarcted myocardium 7 days after MI) and control group (equal volume phosphate buffered saline myocardial injection, n = 8 each). Four weeks after MPM/alpha-CD implantation, echocardiography, histochemistry and immunohistochemistry were performed. RESULTS: Left ventricle ejection fraction was significantly improved in the hydrogel-treated group (56.1% +/- 8.4%) than that in the control group (37.3% +/- 6.4%, P < 0.05). Histological analysis indicated that hydrogel degraded 4 weeks after hydrogel injection, and prevented scar expansion and wall thinning [(3.08 +/- 0.32) mm vs. (2.18 +/- 0.46) mm in control group, P < 0.05]. Neovasculature formation was similar between the hydrogel group [(100.8 +/- 2.4)/mm(2)] and the control [(98.5 +/- 2.9)/mm(2), P > 0.05]. CONCLUSION: MPM/alpha-CD could serve as an excellent injectable biomaterial for improves cardiac function and attenuating scar expansion and left ventricular dilation in MI rabbits.

[Curcumin attenuates left ventricular dysfunction and remodeling in rabbits with chronic heart failure.].

OBJECTIVE: To investigate the effects of Curcumin on rabbits with chronic heart failure. METHODS: Heart failure was induced by combined aortic regurgitation and aortic stenosis in 20 New Zealand rabbits and treated with placebo (HF, n = 10) and Curcumin (Cur, 100 mgxkg(-1)xd(-1), n = 10) for 8 weeks, 10 sham operated rabbits served as controls (Con). Echocardiography was performed in all rabbits at baseline and 8 weeks later. Aortic diameter (AO), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), left ventricular end-systolic dimension (LVDs), left ventricular end-diastolic dimension (LVDd), left ventricular posterior wall thickness (LVPW) and interventricular septum thickness (IVS) were measured. Myocardial matrix metalloproteinase (MMP)-2 and MMP-9 expressions and fibrosis were determined by immunohistochemistry and Masson staining respectively. RESULTS: Compared to baseline, LVEF and LVFS were significantly decreased, AO, LVDs, LVDd, LVPW, and IVS significantly increased at 8 weeks after operation in HF group while these changes could be significantly attenuated in Curcumin treated rabbits. The protein expressions of MMP-2 and MMP-9 were significantly down-regulated in HF group and could be significantly up-regulated by Curcumin treatment. The increased collagen deposition in HF group was also significantly reduced by Curcumin treatment. CONCLUSION: Curcumin attenuated left ventricular dysfunction and remodeling by up-regulating MMPs expressions and reducing myocardial fibrosis.

[Immunogenicity of p55 gene fragment from Pneumocystis carinii].

OBJECTIVE: To evaluate immunogenicity of the recombinant protein GST-p55/570 of Pneumocystis carinii. METHODS: The fusion protein GST-p55/570 was expressed from the prokaryotic expression plasmid pGEX-570, and purified by using glutathione-agarose. The expressed product was analyzed by SDS-PAGE. Thirty-three mice were randomly divided into three groups, immunized with GST-p55/570, GST and PBS, respectively. Each group was immunized for four times at 2 week intervals. At the 7th day after final immunization, spleen was removed to obtain single cell suspension. Proliferation ability of lymphocytes was determined by MTT. Serum samples were collected at pre-immunization and two weeks after each immunization. Antibody level in sera of mice was determined by ELISA. The immune response to the recombinant GST-p55/570 recognized by sera of immunized mice was examined by Western blotting. RESULTS: The expressed fusion protein GST-p55/570 showed a Mr 47,000. Compared with GST group (1.134 5 +/- 0.073 5) or PBS group (1.124 8 +/- 0.041 6), a higher stimulation index (2.063 0 +/- 0.1602) was revealed in GST-p55/570-immunized mice (P < 0.01). At the 14th to 49th day after immunization, the antibody titer against GST-p55/570 in the immunized group was significantly higher than that of GST or PBS groups (P < 0.01). Western blotting indicated that the fusion protein (GST-p55/ 570) had specific immune response to positive serum. CONCLUSION: The fusion protein GST-p55/570 elicits significant humoral and cellular immune responses.

The value of the left atrial appendage orifice perimeter of 3D model based on 3D TEE data in the choice of device size of LAmbre™ occluder.

Preoperative optimal selection of the occluder size is crucial in percutaneous left atrial appendage (LAA) occlusion, and the maximal width of the LAA orifice is the main reference index, however it can not fully meet the practical operation requirements. We retrospectively analyzed three-dimensional (3D) transesophageal echocardiography (TEE) and computed tomography (CT) imaging dataset of the 41 patients who underwent LAA occlusion with LAmbre™ system. The LAA orifice parameters were overall evaluated to determine their role in device size selection. Eight LAA 3D models of the four cases who had been replaced their device during the procedure based on TEE and CT were printed out to verify the optimal parameter decision strategy. There was a significant concordance of the results between 3D TEE and CT in the LAA orifice evaluation. The correlations between the perimeter and maximal width measurements by 3D TEE and the closure disk of the device were stronger than that between the area measurements and the closure disk (r = 0.93, 0.95, 0.86, respectively and p < 0.001 all), and the result was similar to that by CT (r = 0.92, 0.93, 0.84, respectively and p < 0.001 all). The ratios of the maximal width to the minimal width of the four cases were all > 1.4, however the rest 37 cases were all ≤ 1.4. Based on the comprehensive assessment of the LAA orifice perimeter and maximal width of the 3D printed models, the experiments were all succeed just for one try. The LAA orifice perimeter of 3D printed model based on 3D TEE may help in choosing the optimal device size of LAmbre™, especially for the LAA with flater ostial shape.

The implementation of the elastography score in combination with ultrasound prevents unnecessary biopsy of cardiac lesions.

The pathological technique is time consuming, costly, and patients are not preferred routinely. Histopathological findings have very low affectability and greater occurrence of ß-errors, specifically in the diagnosis of cardiomyopathies. Angiography provides a two-dimensional view only. Vascular ultrasound elastography is a comparatively simple diagnostic method with a high resolution of images. The objective of this study was to compare the accuracy of ultrasound, followed by elastography with coronary angiography and endomyocardial biopsy, in the diagnosis of cardiovascular diseases in a Chinese population. A total of 792, patients pathologically abnormal (study group, n=396) and normal (non-study group, n=396), respectively, were included in the experimental diagnostic study. The patients were diagnosed by coronary angiography, endomyocardial biopsy of cardiac lesions, and the Lagrangian speckle model estimator implementation followed by elastography. The study group patients were observed for 38 months after diagnosis. The Mann-Whitney U test followed by Dunnett's multiple comparisons test was used to compare histopathological findings and elastic modulus values between study group and non-study group subjects at a 99% of confidence level. Pathology did reveal a significant cardiac abnormality in the study group patients at baseline. In the angiogram, indistinguishable differences between two distinct parts of the artery were reported. However, the ultrasound images were showed an obvious change in the diameter of the artery for the study group patients (p<0.0001, q=34.301). The histopathological findings were failed to detect a cardiac abnormality in the study group (p=0.0426). However, a significant a cardiac abnormality was observed in elastic modulus values in the study group (p<0.0001 q=4.121). During follow-up, physicians were detected significant cardiovascular diseases in study group patients. Vascular ultrasound elastography is a non-invasive method of diagnostic technique and can increase the confidence of the diagnosis in cases of cardiovascular diseases.

[Construction and expression of prokaryotic expression plasmids of Pneumocystis carinii p55 antigen gene fragment].

OBJECTIVE: To construct prokaryotic recombinant expression plasmid carrying Pneumocystis carinii Mr 55 000 antigen (p55) gene fragment and express the recombinant protein. METHODS: P. carinii pneumonia (PcP) rat models were established by subcutaneous injection of dexamethasone for 14 weeks. Total RNA was extracted from lung of P. carinii rat and p55 antigen gene fragment was cloned by RT-PCR, which was identified by sequencing. The 690 bp fragment was cloned to pGEX-4T-1, the recombinant plasmid was screened and identified by restriction analysis and PCR. The recombinant plasmid was finally induced with IPTG to express a new fusion protein, and the products were analyzed by SDS-PAGE and Western blot. RESULTS: A fragment of 690 bp was obtained by RT-PCR. The recombinant pGEX-4T-1/690 was constructed. SDS-PAGE revealed that the molecular weight of the recombinant protein was approximately Mr 62,000, the maximum amount of the fusion protein produced was 11.6% of the total protein. The recombinant protein can be recognized by GST antibody and by the sera from P. carinii infected rats using Western blotting. CONCLUSION: Prokaryotic expression plasmid pGEX-4T-1/690 has been constructed and the recombinant fusion protein shows antigenicity.

Enhanced effect of nuclear localization signal peptide during ultrasound­targeted microbubble destruction­mediated gene transfection.

Ultrasound­targeted microbubble destruction (UTMD) can promote the entry of plasmid DNA (pDNA) into the cell cytoplasm, by increasing the permeability of the cell membrane. But the transfection efficiency remains low due to inability of the pDNA to enter the nucleus. Various methods have been explored to improve the UTMD transfection efficiency, but with little success. In cells, the classic nuclear localization signal (cNLS) peptide is an amino acid sequence that signals proteins that are due for nuclear transport. The present study aimed to investigate whether binding of a cNLS peptide to the pDNA may improve the transfection efficiency of UTMD. Four experimental groups were analyzed: Control group (UTMD + pDNA), group with cNLS (UTMD + pDNA + cNLS), group with mutated NLS (mNLS; UTMD + pDNA + mNLS), and group with cNLS and the nuclear import blocker, wheat germ agglutinin (WGA; UTMD + pDNA + cNLS + WGA). The NLS was labeled by fluorescein isothiocyanate, whereas pDNA was labeled with Cy3. Different molar ratios were tested for the NLS and pDNA combination in order to achieve optimal binding of the two molecules. Human umbilical vein endothelial cells were then transfected using the optimum ultrasonic irradiation parameters and NLS/pDNA molar ratio. At 6 h post­transfection, the rates of Cy3­labeled pDNA inside the cells and their nuclei were detected by flow cytometry and laser confocal microscopy, and the cellular vs. nuclear uptake of pDNA was calculated. In order to further evaluate the effect of NLS on UTMD­mediated gene transfection, the transfection efficiency and relative expression levels of mRNA and protein were detected at 48 h post­transfection. The results demonstrated that the optimal molar ratio of NLS with pDNA was 104:1. The rates of pDNA successful entry into the cell and nucleus were significantly higher in the cNLS group compared with the control group. The transfection efficiency, and relative expression levels of mRNA and protein from the plasmid were significantly increased in the cNLS group compared with the control group. The mNLS group displayed no significant difference compared with the control group, while the WGA group exhibited significant inhibition in most indicators of transfection efficiency compared to the cNLS group. These results suggest that combining a cNLS peptide with pDNA during UTMD­mediated transfection significantly improved transfection efficiency. Thus, a cNLS peptide may be an important mediator and a new strategy in enhancing the efficiency of UTMD­mediated gene transfection.