In vivo visualization of bonghan ducts inside blood vessels of mice by using an Alcian blue staining method.

An in vivo method using Alcian blue (AB) was developed for visualizing floating threadlike tissues inside blood vessels of mice. These novel structures called intravascular Bonghan ducts (IBHDs) are considered as extension of acupuncture meridians. For in vivo imaging of IBHDs, AB solution (pH 7.4) that stains mucopolysaccharides like hyaluronic acid was used. After injecting AB solution into the femoral vein of a mouse, the threadlike structures, stained deep blue, inside the inferior vena cava. The histological results, such as hematoxylin, eosin, and AB staining, show the compositions of the cells and the extracellular matrix in the IBHD. Further studies are needed to investigate their physiological functions, especially in relation with those of other circulatory systems.

Effect of caspase inhibitors on the post-thaw motility, and integrity of acrosome and plasma membrane of cryopreserved equine spermatozoa.

The present study was designed to test the hypothesis that addition of anticaspase cocktails (inhibiting caspases and thus blocking apoptosis) to the extenders increases the post-thaw viability of equine spermatozoa. The addition of caspase inhibitors failed to improve the acrosome and plasma membrane integrity of spermatozoa, suggesting that in equine sperm cryopreservation protocols, the addition of these caspase inhibitors to cryopreservation medium may not be beneficial in protecting the sperm from the stress of cryopreservation.

Effect of Ketamine on Apoptosis by Energy Deprivation in Astroglioma Cells using Flow Cytometry System

Apoptosis is a programmed, physiologic mode of cell death that plays an important role in tissue homeostasis. As for the central nervous system, ischemic insults can induce pathophysiologic cascade of apoptosis in neurophils. Impairment of astroctye functions during brain ischemia can critically influence neuron survival by neuronglia interactions. We aimed to elucidate the protective effect of ketamine on apoptosis by energy deprivation in astrocytes. Ischemic insults was induced with iodoacetate/ carbonylcyanide mchlorophenylhydrazone (IAA/CCCP) 1.5 mM/ 20 micrometer or 150 micrometer/2 micrometer for 1 hr in the HTB-15 and CRL-1690 astrocytoma cells. Then these cells were reperfused with normal media or ketamine (0.1 mM) containing media for 1 hr or 24 hr. FITC-annexin-V staining and propidium iodide binding were determined by using flow cytometry. Cell size and granularity were measured by forward and side light scattering properties of flow cytometry system, respectively. An addition of keta-mine during reperfusion increased the proportion of viable cells. Ketamine alleviated cell shrinkage and increased granularity during the early period, and ameliorated cell swelling during the late reperfusion period. Ketamine may have a valuable effect on amelioration of early and late apoptosis in the astrocytoma cells, even though the exact mechanism remains to be verified.