PURPOSE: To determine the concentration at which a mixed injection of tissue plasminogen activator (tPA) and C3F8 gas is toxic, we studied the histopathological changes in the rabbit retina. METHODS: Only tPA was injected into the right vitreous cavities of 18 normal pigmented rabbits at doses of 25 micro gram/0.1mL, 50 micro gram/0.1mL, and 100 micro gram/0.1mL, 6 rabbits per dosage. In the same rabbits, tPA and C3F8 (0.2cc) were simultaneously injected into the left vitreous cavities at doses of 25 micro gram/0.1mL, 50 micro gram/0.1mL, and 100 micro gram/0.1mL. All of the eyes were examined by slit lamp biomicroscopy and indirect ophthalmoscopy at 5, 10, and 15 days after the injection, and then they were enucleated for histopathological evaluation. RESULTS: Retinal pigmentary alterations were centered around the injection site 3 days postoperatively in the eyes receiving doses of 50 micro gram/0.1mL or greater. On light microscopy(LM), the involved areas showed vacuolization in the photoreceptor elements and the inner nuclear layer(INL) at a dose of 25 micro gram/0.1mL at postoperative 5 days and the vacuolar changes disappeared at postoperative 15 days. But at doses of 50 micro gram/0.1mL or greater, loss, contracture, and vacuolization of the photoreceptor outer segment (POS) and vacuolization of INL were noted at postoperative 15 days. On LM, at a dose of 25 micro gram/0.1mL, the involved areas showed vacuolization in POS and mitochondrial swelling of the photoreceptor inner segment (PIS) at postoperative 5 days. The mitochondrial swelling of PIS disappeared at postoperative 15 days. However, at doses of 50 micro gram/0.1mL or greater, loss and contracture of POS and mitochondrial swelling of PIS were noted at postoperative 15 days. The retinal damage from simultaneous injection of tPA and C3F8 at doses of 25, and 50 micro gram/0.1mL was equal to or less than that of only tPA injection, whereas at a doses of 100 micro gram/0.1mL the damage was greater. CONCLUSIONS: At doses of 50 micro gram/0.1mL or greater, irreVersible retinal toxicity was noted histopathologically in rabbit eyes. At doses of 25, and 50 micro gram/0.1mL, the degree of retianl damage did not seem to be affected by whether C3F8 was injected concomitantly or not.
Contracture ; Mitochondrial Swelling ; Ophthalmoscopy ; Rabbits ; Retina ; Retinaldehyde ; Tissue Plasminogen Activator*

PURPOSE: This experimental study investigates the effect of mitomycin C(MMC) on the healing of sinus mucosal opening. METHODS: One hundred and twenty rabbits were used and divided into 8 groups of fifteen rabbits. A small(1.35 mm) or large(2.7 mm) diameter mucosal opening was created by drilling in the medial wall of the left maxillary sinus of rabbits and treated with MMC at a concentration of 0.2 or 2.0 mg/ml for 5 or 30 minutes. Five rabbits from each group were sacrificed at 1,2 & 4 weeks. The size of a remained mucosal opening was measured and light and electron microscopic examination were performed. RESULTS: After comparing the sizes of remained mucosal openings among the small opening groups(group 1,3 & 5), group 5 maintained larger opening at 2 weeks than the control group(P<0.05). Light microscopy of the specimens showed inflammatory and degenerative changes. The bone of the maxillary sinus exhibited minimal change induced by MMC. Scanning electron micrographs showed heterochromatin in nuclei and mitochondrial swelling of the epithelium. CONCLUSIONS: The MMC was effective in maintaining a larger mucosal opening than control group at least for 2 weeks following surgery.
Epithelium ; Heterochromatin ; Maxillary Sinus ; Microscopy ; Mitochondrial Swelling ; Mitomycin* ; Rabbits*

PURPOSE: This study aimed to elucidate the molecular mechanisms of the anti-pancreatic fibrosis effects of matrine in rats. MATERIALS AND METHODS: Trinitrobenzene sulfonic acid was administrated to rats to establish a pancreatic fibrosis model. Rats were divided into four groups: Control, Sham, Model, and Matrine (n=8). Hematoxylin-eosin staining, Masson staining, and Azan staining were performed to evaluate pancreatic fibrosis. Expression of transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), and collagen I in pancreatic tissues was evaluated by immunohistochemical staining. mRNA and protein levels of TGF-β receptor 1 (TβR1), TβR2, and Smad2 in pancreatic tissues were determined by RT-PCR and Western blot, respectively. RESULTS: In the model group, hyperplasia of glandules around the glandular ducts, mitochondrial swelling of acinous cells, and severe fibrosis were found. Interestingly, in the Matrine group, mitochondrial swelling was only found in a small number of acinous cells, and the fundamental structures of pancreatic tissues were intact. Moreover, pancreatic fibrosis was markedly alleviated. Comparing to the Sham group, expression of α-SMA, TGF-β1, and collagen I was sharply elevated in the Model group (p < 0.05); however, their expressions were much lower in the Matrine group, compared to the Model group (p < 0.05). Compared with the Sham group, mRNA and protein levels of Smad2, TβR1, and TβR2 in the Model group were notably raised (p < 0.05). However, their high expression was significantly downregulated in the Matrine group (p < 0.05). CONCLUSION: Matrine suppressed pancreatic fibrosis by regulating TGF-β/Smad signaling in rats.
Acinar Cells ; Actins ; Animals ; Blotting, Western ; Collagen ; Fibrosis* ; Hyperplasia ; Mitochondrial Swelling ; Rats* ; RNA, Messenger ; Signal Transduction

PURPOSE: Cell-in-cell structures are created by one living cell entering another homotypic or heterotypic living cell, which usually leads to the death of the internalized cell, specifically through caspase-dependent cell death (emperitosis) or lysosome-dependent cell death (entosis). Although entosis has attracted great attention, its occurrence is controversial, because one cell line used in its study (MCF-7) is deficient in caspase-3. METHODS: We investigated this issue using MCF-7 and A431 cell lines, which often display cell-in-cell invasion, and have different levels of caspase-3 expression. Cell-in-cell death morphology, microstructures, and signaling pathways were compared in the two cell lines. RESULTS: Our results confirmed that MCF-7 cells are caspase-3 deficient with a partial deletion in the CASP-3 gene. These cells underwent cell death that lacked typical apoptotic properties after staurosporine treatment, whereas caspase-3-sufficient A431 cells displayed typical apoptosis. The presence of caspase-3 was related neither to the lysosome-dependent nor to the caspase-dependent cell-in-cell death pathway. However, the existence of caspase-3 was associated with a switch from lysosome-dependent cell-in-cell death to the apoptotic cell-in-cell death pathway during entosis. Moreover, cellular hypoxia, mitochondrial swelling, release of cytochrome C, and autophagy were observed in internalized cells during entosis. CONCLUSION: The occurrence of caspase-independent entosis is not a cell-specific process. In addition, entosis actually represents a cellular self-repair system, functioning through autophagy, to degrade damaged mitochondria resulting from cellular hypoxia in cell-in-cell structures. However, sustained autophagy-associated signal activation, without reduction in cellular hypoxia, eventually leads to lysosome-dependent intracellular cell death.
Apoptosis ; Autophagy ; Caspase 3* ; Cell Death ; Cell Hypoxia ; Cell Line ; Cytochromes c ; Entosis ; MCF-7 Cells* ; Mitochondria ; Mitochondrial Swelling ; Staurosporine

Thalidomide, a potent teratogen, is Known as an angiogenic inhibitor. This study was performed to examine the effect of thalidomide on corneal angiogenesis in rabbit cornea induced by chemical cauterization. We applied Whatman filter paper disc soaked in 30% silver nitrate (AgNO3) application on corneas of 12 white rabbits. After 5days, we administered oral dose of 100mm2 thalidomide to the 6 animals everyday and examined the length and extent of neovascularization to evaluate the area of neovascularization. After 2 days of oral administration, the increase of neovascularization is 14.3+/-11.7mm2in thalidomide-treated group and 27.9+/-14.6mm2 in cotrol grop. The area of neovascularization reached to its maximum at day 9 in thalidomide-treated group compared to day 11 in control group and decreased thereafter in both groups. The increase of the area of vascularized cornea revealed 28.0+/-13.5mm2 in thalidomide-treated group and 44.4+/-12.7mm2 in control group at the day 9 (p=0.04, Wilkoxon Matched-pairs signed-rank test). This fact means that treatment with thalidomide resulted in an inhibition of the area of vascularized cornea with the median inhibition of 37.3%. On light micrographs, there were infiltration of inflammatory cell and capillary lumens in corneal stroma in both animals. Electron micrographs of thalidomide-treated animals showed loss of vascular endothelial cell junction, mitochondrial swelling and loss of cristae which were not found in control animals. This results suggest that orally-administered thalidomide has a direct effect on the growing vasculature and an inhibitory effect on corneal angiogenesis.
Administration, Oral ; Animals ; Capillaries ; Cautery ; Cornea ; Corneal Neovascularization* ; Corneal Stroma ; Endothelial Cells ; Mitochondrial Swelling ; Rabbits ; Silver Nitrate ; Thalidomide*

OBJECTIVESTo investigate the effects of infrasound on ultrastructure of testis in mouse.

METHODSTwelve male BALB/C mice were randomly divided into three groups according to exposed duration on 1, 7 and 14 day. The mice were separately exposed to infrasound environment under 8 Hz/90 dB, 8 Hz/130 dB, 16 Hz/90 dB, 16 Hz/130 dB 2 hours per day. There was another control group which had three mice were separated into module with no infrasound. All the mice were killed on schedule. Then all the sections of testis were observed under electronic microscope. The alterations of structure and the chromatin were observed.

RESULTSSome acute alteration in one day group was found in testis cell, such as cellular denaturation and necrosis, intercellular edema, mitochondria swelling, liposome hyperplasia. When the infrasound was up to 8 Hz/130 dB, the damage showed seriously. In 7 and 14 day group, the acute alteration was gradually decreased. A plenty of abnormal sperm were found. And other alteration was chromatin condense. The effect of variational frequency was important in ultrastructure.

CONCLUSIONSThe infrasound markedly effected to testicular cell morphology and secreting function. Infrasound will lead to the alteration of procreation in mouse.

Animals ; Cell Size ; Male ; Mice ; Mice, Inbred BALB C ; Mitochondrial Swelling ; Sound ; adverse effects ; Testis ; secretion ; ultrastructure

OBJECTIVETo investigate the effects of resveratrol (Res) on mitochondrial opening and Ca(2+)-induced Ca(2+) release (CICR) from rat liver cell mitochondria mediated by Ca(2+).

METHODSWistar rat liver cell mitochondria was extracted and Res-induced mitochondrial swelling was assessed spectrophotometrically at 540 nm to examine the permeability transition pore (PTP) opening. The membrane potential changes of Res-treated mitochondria were measured with fluorescence spectrophotometery. Ca(2+) uptake and release by the mitochondria was determined by absorbance change of arsenazo III at 685-675 nm monitored by dual wavelength spectrophotometry.

RESULTSRes promoted Ca(2+)-mediated PTP opening, and this effect was completely inhibited by CsA and lowered by trifluoperazine. CICR accelerated by Res treatment was completely blocked by ruthenium red and partly by trifluoperazine.

CONCLUSIONRes can promote PTP opening by inducing CICR, which may be one of the pathways that Res induces cell apoptosis.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Female ; Hepatocytes ; cytology ; metabolism ; Mitochondria, Liver ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; metabolism ; Mitochondrial Swelling ; drug effects ; Rats ; Rats, Wistar ; Stilbenes ; pharmacology

OBJECTIVETo study effects of three pentacyclic triterpenoids, oleanolic acid (OA), ursolic acid (UA) and asiatic acid (AA) on Ca(2+)-induced liver mitochondrial permeability transition (MPT).

METHODEffects of three compounds on liver MPT induced by Ca2+ were assessed by measuring the change in mitochondrial swelling, mitochondrial membrane potential and release of matrix Ca2+ in vitro.

RESULTObvious mitochondrial swelling, loss of mitochondrial membrane potential and release of matrix Ca2+ occurred after the addition of 50 micromol x L(-1) Ca2+. However, preincubation with 50 mg x L(-1) OA, UA or AA significantly blocked the above changes. In addition, it was also found that there are differences in the inhibitions of three compounds on liver MPT induced by Ca2+.

CONCLUSIONThree pentacyclic triterpenoids, OA, UA and AA, have significant mitochondrial protection through blocking on liver MPT and the inhibition on liver MPT of AA is stronger than that of UA and OA.

Animals ; Calcium ; metabolism ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Inbred ICR ; Mitochondria, Liver ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; drug effects ; Mitochondrial Swelling ; drug effects ; Pentacyclic Triterpenes ; pharmacology

Effects of aconitine on the retina and optic nerve were studied in rabbits by means of visual evoked potential(VEP), electroretinogram(ERG), and electron microscopy. Aconitine(0.4mg/kg) was administered intra peritoneally. The effects of aconitine were observed at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, and 3 months after injection, respectively. The amplitude of VEP was decreased significantly at 1 and 2 weeks after injection. However ERG b-wave, c-wave, and the latency of VEP were not significantly changed during the experimental periods. Histopathological changes in the optic nerve and visual streak were characterized by disorganization of the lamellar structures of the myelin sheath, diffuse mitochondrial swelling, and vacuolization in the myelinated nerve fibers, which were increased in degree with time. No perceptible change in the retina including retinal pigment epithelium was observed in the experimental periods. Schwann cells and unmyelinated nerve fibers appeared unaffected, except for mild swelling of mitochondria. Above mentioned electrophysiological and electron microscopic studies indicated that the effect of aconitine on the rabbit eye was toxic myelo-optic neuropathy.
Aconitine* ; Microscopy, Electron ; Mitochondria ; Mitochondrial Swelling ; Myelin Sheath ; Nerve Fibers, Myelinated ; Nerve Fibers, Unmyelinated ; Optic Nerve ; Optic Nerve Diseases* ; Rabbits ; Retina ; Retinal Pigment Epithelium ; Schwann Cells

The author studied the functional derangement of blood-retinal barrier induced by destruction of the pigment epithelial cells of the retina. Sodium iodate, which was well known to exert a selectively destructive action to the retinal pigment epithelial cells, was injected to the rabbits intravenously(60mg/kg of body weight). Eyes were enucleated 2 days and 4 days after sodium iodate injection and examined by electron microscope. Some of the tissue were fixed in colloid lanthanum, to investigate the changes of the permeability of plasma membrane in accordance with cellular damages induced by sodium iodate. The permeability of the blood-retinal barrier was also studied after intravenous(200mg/kg) or intraocular(4 microgram/20ml of saline) injection of horseradish peroxidase(HRP). The results obtained were summarized as the following: Sodium iodate induced patchy areas of loss of pigment epithelial cells after 2 days, which were more widespread and severe after 4 days with regenerative activities. Loss of outer segment and mitochondrial swelling of the inner segment of visual cells were also noted after 4 days. Colloidal lanthanum penetrated into the mitochondria of pigment epithelial cells at 2 days after sodium iodate injection, which was extended to the mitochondria of inner segment of visual cells after 4 days. Intraocularly injected HRP appeared from the internal limiting membrane to Bruch's membrane after 2 days. Intravenously injected HRP appeared from the Bruch's membrane to ganglion cell layer after 2 days, which were extended to the vitreal cavity. The results suggested that the damage of the pigment epithelial cells induced by sodium iodate destroy blood-retinal barrier. HRP exudation is more extensive in direction of retina to choroid than choroid to retina.
Armoracia ; Blood-Retinal Barrier* ; Bruch Membrane ; Cell Membrane ; Choroid ; Colloids ; Epithelial Cells ; Ganglion Cysts ; Lanthanum ; Membranes ; Mitochondria ; Mitochondrial Swelling ; Permeability ; Rabbits ; Retina ; Retinal Pigment Epithelium* ; Retinaldehyde* ; Sodium