Mechanically sensitive HSF1 is a key regulator of left-right symmetry breaking in zebrafish embryos.

The left-right symmetry breaking of vertebrate embryos requires nodal flow. However, the molecular mechanisms that mediate the asymmetric gene expression regulation under nodal flow remain elusive. Here, we report that heat shock factor 1 (HSF1) is asymmetrically activated in the Kupffer's vesicle of zebrafish embryos in the presence of nodal flow. Deficiency in HSF1 expression caused a significant situs inversus and disrupted gene expression asymmetry of nodal signaling proteins in zebrafish embryos. Further studies demonstrated that HSF1 is a mechanosensitive protein. The mechanical sensation ability of HSF1 is conserved in a variety of mechanical stimuli in different cell types. Moreover, cilia and Ca2+-Akt signaling axis are essential for the activation of HSF1 under mechanical stress in vitro and in vivo. Considering the conserved expression of HSF1 in organisms, these findings unveil a fundamental mechanism of gene expression regulation by mechanical clues during embryonic development and other physiological and pathological transformations.

Comparative study of BRAF gene mutations in ovarian serous tumors by immunohistochemistry and DNA sequencing.

PURPOSE: Somatic mutations in the BRAF gene are common in several types of cancer, especially in ovarian serous cancer (OSC). Normally, the BRAF protein is switched on and off in response to signals that control cell growth and development. METHODS: To investigate the correlation between the mutation of BRAF gene and the expression of BRAF protein in OSC, pyrosequencing was performed to detect the mutation of the 600th codon in BRAF gene (written as Val600Glu or V600E) in 23 cases of high-grade serous ovarian cancer (HGSC), 28 cases of low-grade serous ovarian cancer (LGSC) and 72 cases of serous borderline ovarian tumors (SBT). Meanwhile, immunohistochemistry which stained with the specific antibody VE1 were used to clarified the expression level of BRAF V600E mutant protein. RESULTS: Finally, we found that V600E mutation in LGSC and SBT of occurred in 2 of 23 (7.1%) and 21of 72 (29.2%), respectively. The V600E mutation was not detected in 23 cases of HGSC. One case of HGSC (1, 4.3%), 3 cases of LGSC (3 of 28, 10.7%) and 25 cases of SBT (25 of 72, 34.7%) were positive expression detected by immunohistochemistry. Compared with BRAF gene mutation, the sensitivity, specificity and consistency of BRAF V600E protein were 91.3%, 92% and 91.9%, respectively. CONCLUSION: Our findings indicate that BRAF mutations in LGSC and SBT, which are closely related to tumor staging. The specific antibody VE1 could be used as a preliminary screening for the mutation of BRAF gene.

The diagnosis and treatment of cardiac lymphangioma: A case report and literature review.

RATIONALE: Cardiac lymphangioma is a rare disease. Until now, there have been only a few cases of cardiac lymphangioma reported in the literature. PATIENT CONCERNS: We report the case of a 57-year-old female patient with cardiac lymphangioma from atrial septum. DIAGNOSIS: Color Doppler echocardiography was performed, which revealed a tumor occupying a large amount of space in the left and right atrium. INTERVENTIONS: The patient underwent thoracoscopic cardiac tumor resection under general anesthesia according to the procedure used for benign tumors. OUTCOMES: The patient recovered completely and was discharged home. Follow-up color Doppler echocardiography scans obtained from 6 months to 2 years after the operation showed no recurrent mass. LESSONS: Once the tumor is detected, surgical treatment should be implemented as soon as possible.

The transcription factor Ikaros inhibits cell proliferation by downregulating ANXA4 expression in hepatocellular carcinoma.

The occurrence and progression of hepatocellular carcinoma (HCC) are affected by complicated signal transduction factors. Our previous study identified Ikaros as a novel reactivated therapeutic target that acts as a transcriptional repressor and reactivates anticancer mechanisms in HCC therapy. Annexin A4 (ANXA4) is a member of the Annexin family that plays an essential role in several cancers, but it has not been investigated in HCC proliferation. Using cDNA microarrays, ANXA4 was shown to be associated with Ikaros in Ikaros-overexpressing cells. The aim of this work was to characterize the relationship between Ikaros and ANXA4 and the role of ANXA4 in HCC. The effect of Ikaros on ANXA4 was analyzed in HCC cell lines and HCC patient samples, and functional recovery experiments were performed between Ikaros and ANXA4. Furthermore, the effect of ANXA4 on cell proliferation in vitro was analyzed by MTT and colony formation assays in HCC cells. We used a subcutaneous xenograft model to elucidate the role of ANXA4 in vivo. We found that ANXA4 overexpression promotes HCC cell proliferation, but Ikaros can inhibit ANXA4 expression by repressing its promoter activity. Moreover, we demonstrated that downregulated expression of ANXA4 inhibited HCC cell proliferation and tumorigenesis in vitro and in vivo. Our findings indicate that ANXA4 may be a critical factor in HCC tumorigenesis. Ikaros is an attractive inhibitor of ANXA4 and may function as an anticancer agent in HCC.

[Optical Analysis of the Interaction of Mercaptan Derivatives of Nanogold Particles with Carcinoembryonic Antigen].

Gold nanoparticles (AuNPs) have been the subject of intense research for use in biomedicine over the past couple of decades. AuNPs, also referred to as colloidal gold, possess some astounding optical and physical properties that have earned them a prime spot among the new promising tools for medical applications. Today, AuNPs are offered to provide the clinical laboratory with more sensitive, faster, and simpler assays, which are also cost-effective. AuNPs can be used to develop point-of- care tests and novel testing strategies such as in drug targeting, disease detection, molecular recognition, and biological labels. The typical structure of AuNPs is spherical nano-sized gold particles, but they can also be composed of a thin gold shell surrounding a dielectric core, such as silica (gold nanoshells). their size range from 0.8 to 250 nm and are characterized by high absorption coefficients. AuNPs have some unique optical properties, such as enhanced absorption and scattering, where the absorption cross-section of AuNPs is 4~5 orders of magnitude greater than that of rhodamine 6G. When AuNPs aggregate, interaction of locally adjacent AuNPs (plasmon-plasmon interaction) shifts their color to blue. Thus, the binding of AuNP-labeled entities to their respective target would lead to aggregation of the nanoparticles and a detectable shift in the optical signal. The strong absorption of AuNPs can also be used in colorimetric detection of analytes by measuring changes in the refractive index of the AuNP's environment caused by adsorption of the target analytes. However, a large number of surface atoms of nanoparticles have huge surplus bonding ability, because of surface effect of gold nanoparticles, result in reuniting and sinking among the nanoparticles which make them unstable. In order to detect traces of carcinoembryonic antigen, one of the tumor targets, a new kind of gold nanoparticle with hyperchormic effect and fluorescence sensitization effect material needs to be prepared. In this paper, novel mercaptan derivative of nanogold particles are prepared and studied using transmission electron microscopy (TEM), ultra-violet-visible absorption spectra (UV-Vis), fluorescence emission (FE) spectrum and infrared spectrum (IR) methods. The UV-Vis and FE results show the presence of new ligands mercaptan, more electrons from the orbit of ligand which can excite to the central ion related orbits and increase fluorescence of gold. Fluorescence sensitization effect was observed when mercaptan derivatives of nanogold interacted with carcinoembryonic antigen (CEA) and no fluorescence sensitization effect was found when nanogold interacted with carcinoembryonic antigen (CEA). The study of CEA hyperchromic mechanism of mercaptan derivatives nanogold and the CEA by the method of infrared spectrum, shows that the randomized OH bonds in the Au-protein interaction, showed more on the outside of the plane of bending vibration after the interaction with the mercaptan derivative nanogold, making the energy transfer from mercaptan derivatives nanogold to protein easy; leading to its fluorescence sensitization effect.

Green light-emitting polyepinephrine-based fluorescent organic dots and its application in intracellular metal ions sensing.

In this paper, we present a class of bio-dots, polyepinephrine (PEP)-based fluorescent organic dots (PEP-FODs) for selective and sensitive detection of Fe(2+), Fe(3+), and Cu(2+). The PEP-FODs were derived from epinephrine via self-polymerization at relatively low temperature down to 60°C with low cytotoxicity and relative long lifetime (7.24ns). The surface morphology and optical properties of the synthesized PEP-FODs were characterized. We found that the diameters of PEP-FODs were mainly distributed in the narrow range of 2-4nm with an average diameter of 2.9nm. An optimal emission peak located at 490nm was observed when the green light-emitting PEP-FODs were excited at 400nm. It is discovered that Fe(2+), Fe(3+), and Cu(2+)can strongly quench the fluorescence of PEP-FODs through the nonradiative electron-transfer. The detection limit of 0.16, 0.67, and 0.15µM was obtained for Fe(2+), Fe(3+), and Cu(2+), respectively. The independent sensing platform of Fe(2+), Fe(3+), and Cu(2+)could be established by using NaF as a complexing agent and by regulating the reaction time between NaF and metal ions. Cell viability studies reveal that the as-prepared PEP-FODs possess good solubility and biocompatibility, making it as excellent imaging nanoprobes for intracellular Fe(2+), Fe(3+), and Cu(2+)sensing. The developed PEP-FODs might hold great promise to broaden applications in nanotechnology and bioanalysis.

Research on the compatibility laws in treating rheumatoid arthritis patients of cold-dampness obstruction syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To find out Chinese herbal compounds (CHCs) with high frequency in treating rheumatoid arthritis (RA) patients of cold-dampness obstruction syndrome (CDOS) by inductively reviewing literatures concerning clinical application of CHCs, thus improving theoretical and medical practice levels of Chinese medical recipes.</p><p><b>METHODS</b>The CHCs for treating RA patients of CDOS published in medical journals in recent 30 years were retrieved. The database of CHCs was set up. The herbal functions, meridian tropism, flavors and properties were statistically analyzed.</p><p><b>RESULTS</b>There were 126 single herbs in CHCs for treating RA patients of CDOS. The top 31 single herbs occupied 70.7% cumulative frequency, being high frequency CHCs for treating RA patients of CDOS. From the function aspect, the anti-rheumatic herbs, asthenia supplementing herbs, diaphoretic, and herbs for activating blood circulation and removing blood stasis were identified as high frequency CHCs. From the meridian tropism aspect, Gan-meridian, Pi-meridian and Shen-meridian, occupied the top 3. As for flavors, acid, bitter, and sweet occupied the top three. As for drug nature, herbs of warm property were used most frequently.</p><p><b>CONCLUSION</b>The high frequency CHCs obtained from analytical statistics could provide evidence for clinical medication.</p>

[Establishment of a porcine model of congenital heart defect with decreased pulmonary blood flow].

OBJECTIVE: To establish an animal model of congenital heart defect with decreased pulmonary blood flow for better understanding the pathophysiology of pulmonary vascular development and related regulatory mechanisms of congenital heart defect with decreased pulmonary blood flow. METHOD: One to two months old pigs were randomly divided into three groups: control group (group C, n = 6) with right chest small incisions induced transient pulmonary blood reduction; light-moderate stenosis groups (group T(1), n = 7): artificial atrial septum defect (ASD) plus controlled pulmonary artery banding to generate a systolic pressure gradient of 20 - 30 mm Hg (1 mm Hg = 0.133 kPa); severe stenosis groups (group T(2), n = 7): similar surgical procedures as group T(1), and controlled pulmonary artery banding to generate a systolic pressure gradient ≥ 30 - 50 mm Hg. 64-slice computed tomography scanning was performed at one month post operation. Arterial blood gas analysis, hemoglobin value, pulmonary vessel, ASD and banding ring diameters and trans-pulmonary artery banding pressure (Trans-PABP) were determined at two months post operation. RESULTS: One pig died due to tracheal intubation accident in the C group, one pig died due to bowel obstruction in the T(1) group and two pigs died due to acute right heart failure and chronic heart failure respectively in T(2) group. 64-slice CT angiography results showed that aortic diameter of T(1) group was significantly lower than that of C group and banding diameter was significantly lower than aortic diameter in the T(1) and T(2) groups at one month post operation. Two months after operation, the size of ASD were (8.0 ± 0.5) mm and (8.9 ± 1.4) mm (P > 0.05) respectively in the T(1) and T(2) groups after operation. The Trans-PABP was significantly higher in the T(1) and T(2) groups than in C group (P < 0.01), and the Trans-PABP was significantly higher in the T(2) group than in T(1) group (P < 0.01). PaO2 and SaO2 in the T(1) and T(2) groups were significantly lower than those in C group. CONCLUSION: Artificial atrial septum defect combined pulmonary artery banding procedures could be successfully used to establish model of congenital heart defect with decreased pulmonary blood flow and this model could help to understand the pathophysiology and monitor therapy efficacy for patients with congenital heart defect with decreased pulmonary blood flow.

Vascular gene transfer and drug delivery in vitro using low-frequency ultrasound and microbubbles.

AIM: To determine the effects of ultrasound exposure in combination with a microbubble contrast agent (SonoVue) on the cellular uptake and delivery of drugs/genes into human umbilical vein endothelial cells (HUVECs) as well as their biological effects on migration. METHODS: HUVECs in suspension were exposed to pulsed ultrasound with a 10% duty cycle in combination with various concentrations of a microbubble contrast agent (SonoVue) using a digital sonifier at a frequency of 20 kHz and an intensity of 3.77 W/cm(2) on the surface of a horn tip. Cell culture inserts were used to determine the cell migration ability. RESULTS: Exposure to pulsed ultrasound resulted in enhanced green fluorescent protein (EGFP) gene transfection efficiencies ranging from 0.2% to 2%. The transfection efficiency of HUVECs was approximately 3-fold higher in the presence of SonoVue than in its absence at the effective exposure time of 6 s. For drug delivery to HUVECs using ultrasound, the delivery efficiencies of a low-molecular-weight model drug (TO-PRO-1, M(W) 645.38) were significantly higher when compared to drug delivery without ultrasound, with a maximum efficiency of approximately 34%. However, the delivery efficiencies of a high-molecular-weight model drug (Dextran-Rhodamine B, M(W) 70,000) were low, with a maximum delivery efficiency of nearly 0.5%, and gene transfection results were similarly poor. The migration ability of HUVECs exposed to ultrasound was also lower than that of the control (no exposure). CONCLUSION: The use of low-frequency and low-energy ultrasound in combination with microbubbles could be a potent physical method of increasing drug/gene delivery efficiency. This technique is a promising nonviral approach that can be used in cardiovascular disease therapy.