Effect of Salvianolate on Myocardial Ischemia Reperfusion Injury and Its Mechanism

<sec> <title>Background:</title> The paper investigated the H9c2 cardiomyocyte model induced by hypoxia. Cell viability was monitored by real-time unlabeled cell function analyzer to determine the levels of LDH, MDA and SOD in cell supernatant. </sec> <sec> <title>Material and Methods:</title> The cytoskeleton staining was labeled by phalloidin staining. WB was applied to detect the expression of myocardial cytoskeleton microtubuleassociated protein and the expression of HIF-1α protein in each group. After adding AMPK inhibitor Compound C, Hoechst 33342 was employed to detect the apoptosis rate of cardiomyocytes, and WB was applied to detect the expressions of myocardial cytoskeleton-associated protein and p-AMPK. </sec> <sec> <title>Results:</title> Salvianolate can effectively improve cell viability, reduce LDH and MDA levels, increase SOD content, improve skeletal structure damage, reduce nuclear concentration, reduce cell debris, and promote the expressions of microtubule-associated protein, α-tubulin and β-tubulin, MAP4, and microfilament-associated protein MLCK, p-MLC-2 in myocardial cytoskeleton microtubules after ischemia and hypoxia. The addition of AMPK inhibitor can inhibit the expressions of p-AMPK, tubulin MAP4, microfilament protein MLCK and p-MLC-2 up-regulated by Salvianolate. </sec> <sec> <title>Conclusion:</title> Salvianolate can promote the expressions of microtubule-associated protein α-tubulin, β-tubulin,MAP4, microfilament-associated protein MLCK and p-MLC-2 in myocardial cytoskeleton after ischemia and hypoxia, indicating that Salvianolate can protect the myocardial cytoskeleton after ischemia and hypoxia, and may protect the structure and function of microtubules and microfilaments in the myocardial cytoskeleton through the AMPK/MAP4 and AMPK/MLCK pathways. </sec>

Using Flow Cytometry and Multistage Machine Learning to Discover Label-Free Signatures of Algal Lipid Accumulation

Abstract—Most applications of flow cytometry or cell sorting rely on the conjugation of fluorescent dyes to specific biomarkers. However, labeled biomarkers are not always available, they can be costly, and they may disrupt natural cell behavior. Label-free quantification based upon machine learning approaches could help correct these issues, but label replacement strategies can be very difficult to discover when applied labels or other modifications in measurements inadvertently modify intrinsic cell properties. Here we demonstrate a new, but simple approach based upon feature selection and linear regression analyses to integrate statistical information collected from both labeled and unlabeled cell populations and to identify models for accurate label-free single-cell quantification. We verify the method’s accuracy to predict lipid content in algal cells(Picochlorum soloecismus)during a nitrogen starvation and lipid accumulation time course. Our general approach is expected to improve label-free single-cell analysis for other organisms or pathways, where biomarkers are inconvenient, expensive, or disruptive to downstream cellular processes.

Correlative microscopy of colloidal gold-labeled and unlabeled cell-surface-associated molecules: LV-SEM, SEM, and VLM

Video enhanced, interference based light microscopy (VLM) is sufficiently sensitive to permit observation, via the inflated diffraction image, of colloidal gold particles as small as 15nm. These particles can be directly coupled to ligands such that ligand binding, distribution, and ligand-receptor complex movement can be observed on living cells (Fig.1a-d) and correlated subsequently with HVEM and/or SEM images of the same cell (Fig. 1e). The size of the gold particles used in these studies is such that, other than for very large ligands, generally two or more ligand molecules are bound per particle. Thus questions regarding the role of the polyvalent “particle” (ligand-gold conjugate) vs. soluble ligand can arise. Observation of individual labeled or unlabeled ligand molecules can therefore become useful in resolving such questions when they occur.

Spatial geometry of the dopamine innervation in the avascular area of the human fovea

The dopamine (DA) innervation, labeled by tyrosine hydroxylase immunohistochemistry in a wholemounted human retina, is described in the avascular area of the fovea. Eleven DA neurons give rise to this innervation, among which five are interplexiform cells, so that the DA innervation consists of two plexuses: one is internal and is formed by the dendrites of all of the DA cells, and the other is external and is formed by the scleral processes of the interplexiform cells. Five concentric zones are delineated according to the focal plane in which the internal DA plexus is observed. The central zone 1 contains DA processes crossing in all directions. Zones 2 and 3 do not contain any cell bodies. In zone 3 the internal plexus begins to undergo a concentric arrangement, which is clearly observed in zones 4 and 5. The external DA innervation displays a different appearance in zones 1, 2, and 3, in which it consists of vertically oriented thin processes and terminals penetrating the outer nuclear layer, vs. zones 4 and 5 in which it consists of both the same type and horizontal processes lying in the outer plexiform layer. On the basis of DA-innervation appearance and distribution of labeled and unlabeled cell somata, it was concluded that zones 1, 2, and 3 contained the DA innervation of the foveola. DA processes filtering between photoreceptor cells are particularly well-observed in this region. This anatomical study of the DA innervation in the human fovea leads to a better understanding of the important role of DA in primate central vision and can be used as a reference for an approach of macular pathology.

STUDIES ON THE ORIGIN OF RIBOSOMES IN AMOEBA PROTEUS

The origin of cytoplasmic RNA and ribosomes was studied in Amoeba proteus by transplantation of a radioactive nucleus into an unlabeled cell followed by examination of the cytoplasm of the recipient for the presence of label. When a RNA-labeled nucleus was used, label appeared in the ribosomes, ribosomal RNA, and soluble RNA. Since the kinetics of appearance of labeled RNA indicates that the nucleus was not injured during the transfer, and since the transferred nuclear pool of labeled acid-soluble RNA precursors is inadequate to account for the amount of cytoplasmic RNA label, it is concluded that cytoplasmic ribosomal RNA is derived from acid-insoluble nuclear RNA and is probably transported as an intact molecule. Likewise, cytoplasmic soluble RNA probably originated in the nucleus, although labeling by terminal exchange in the cytoplasm is also possible. The results were completely different when a protein-labeled nucleus was grafted into an unlabeled host. In this case, label was found only in soluble proteins in the host cell cytoplasm, and there were no (or very few) radioactive ribosomes. This suggests that the nuclear pool of ribosomal protein and ribosomal protein precursors is relatively small and perhaps nonexistent (and, furthermore, shows that there was no cytoplasmic ribosomal contamination of the transferred nucleus).