OBJECTIVETo study the effect of ligustrazine nanoparticles nano spray (LNNS) on transforming growth factor β (TGF-β)/Smad signal protein of rat peritoneal mesothelial cells (RPMC) induced by tumor necrosis factor α (TNF-α), and the anti-adhesion mechanism of LNNS in the abdominal cavity.

METHODSThe primary culture and subculture of rat peritoneal mesothelial cells (RPMC) was processed by trypsin digestion method in vitro. The third generation was identifified for experiment and divided into 5 groups: a blank group: RPMC without treatment; a control group: RPMC stimulated with TNF-α; RPMC treated by a low-dosage LNNS group (2.5 mg/L); RPMC treated by a medium-dosage LNNS group (5 mg/L); and RPMC treated by a high-dosage LNNS group (10 mg/L). Reverse transcription-polymerase chain reaction was applied to test the expression of fifibronectin, collagen I (COL-I), TGF-β mRNA, and Western blot method to test the Smad protein 7 expression of RPMC.

RESULTSCompared with the blank group, a signifificant elevation in fifibronectin (FN), COL-I and TGF-β mRNA expression of RPMC were observed in the control group (P<0.05). Compared with the control group, LNNS suppressed the expressions of FN, COL-I and TGF-β mRNA in a concentrationdependent manner (P<0.05). The expression of Smad7 protein of RPMC was down-regulated by TNF-α stimulation, and up-regulated with the increase of LNNS dose (P<0.05).

CONCLUSIONSTNF-α may induce changes in RPMC's viability, leading to peritoneal injury. LNNS could reverse the induction of fifibrosis related cytokine FN, COL-I and TGF-β, up-regulating the expression of Smad7 by TNF-α in RPMC, thus attenuate peritoneal injury by repairing mesothelial cells.

Animals ; Collagen Type I ; genetics ; metabolism ; Epithelium ; drug effects ; metabolism ; Fibronectins ; metabolism ; Male ; Nanoparticles ; chemistry ; ultrastructure ; Particle Size ; Peritoneal Cavity ; cytology ; Pyrazines ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVE: To observe the morphological differences between normal uncinate process(UP) mucosa and inferior turbinate mucosa, and explore the physiology function of the UP with the electron microscope. METHOD: The experiment chose 12 patients who have taken nasal endoscopic surgeries(8 cases for normal UP, 4 cases for normal inferior turbinate mucosa). During the surgery, take the mucosa upwards on the filter paper and immediately use scanning electron microscopy and transmission electron microscopy specimens for standard sample preparation methods. Observe the cilia shape, structure and the distribution and the swing direction. RESULT: (1)The internal side and the external side of UP mucosa and inferior turbinate mucosa are all pseudostratified ciliated columnar epithelium, the shapes of cilia are classic "9+2" structures. The distribution of cilia on internal and external lateral of UP and inferior turbinate mucosa are in high density. (2)The direction of cilia on normal inferior turbinate mucosa are generally swing to up and backwards; the cilia on internal lateral of the UP generally swing towards inner side, down and backwards; the cilia on external lateral of the UP generally swing towards down and backwards. CONCLUSION The cilia on internal side and the external side of UP mucosa and inferior turbinate mucosa are in the same structure and shape, but the swing direction of cilia have their own characteristics. It can be concluded that the internal and external lateral of UP may have different functions in nasal sinuses mucus cilia clearance system.
Cilia ; ultrastructure ; Endoscopy ; Epithelium ; ultrastructure ; Humans ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Nasal Mucosa ; ultrastructure ; Nasal Surgical Procedures ; Paranasal Sinuses ; Turbinates ; ultrastructure

This study investigated the toxicity of commercial non-steroid anti-inflammatory drug (NSAID) eye solutions against corneal epithelial cells in vitro. The biologic effects of 1/100-, 1/50-, and 1/10-diluted bromfenac sodium, pranoprofen, diclofenac sodium, and the fluorometholone on corneal epithelial cells were evaluated after 1-, 4-, 12-, and 24-hr of exposure compared to corneal epithelial cell treated with balanced salt solution as control. Cellular metabolic activity, cellular damage, and morphology were assessed. Corneal epithelial cell migration was quantified by the scratch-wound assay. Compared to bromfenac and pranoprofen, the cellular metabolic activity of diclofenac and fluorometholone significantly decreased after 12-hr exposure, which was maintained for 24-hr compared to control. Especially, at 1/10-diluted eye solution for 24-hr exposure, the LDH titers of fluorometholone and diclofenac sodium markedly increased more than those of bromfenac and pranoprofen. In diclofenac sodium, the Na+ concentration was lower and amount of preservatives was higher than other NSAIDs eye solutions tested. However, the K+ and Cl- concentration, pH, and osmolarity were similar for all NSAIDs eye solutions. Bromfenac and pranoprofen significantly promoted cell migration, and restored wound gap after 48-hr exposure, compared with that of diclofenac or fluorometholone. At 1/50-diluted eye solution for 48-hr exposure, the corneal epithelial cellular morphology of diclofenac and fluorometholone induced more damage than that of bromfenac or pranoprofen. Overall, the corneal epithelial cells in bromfenac and pranoprofen NSAID eye solutions are less damaged compared to those in diclofenac, included fluorometholone as steroid eye solution.
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage/*toxicity ; Benzophenones/administration & dosage/toxicity ; Benzopyrans/administration & dosage/toxicity ; Bromobenzenes/administration & dosage/toxicity ; Cell Movement/drug effects ; Cells, Cultured ; Diclofenac/administration & dosage/toxicity ; Epithelial Cells/drug effects/metabolism/ultrastructure ; Epithelium, Corneal/cytology/*drug effects/metabolism ; Fluorometholone/administration & dosage/toxicity ; Humans ; L-Lactate Dehydrogenase/metabolism ; Microscopy, Electron, Transmission ; Ophthalmic Solutions ; Propionates/administration & dosage/toxicity

The role of methyl-CpG binding domain protein 2 (MBD2) in an ApoE-deficient mouse model of age-related macular degeneration (AMD) was investigated. Eight-week-old Mbd2/ApoE double deficient (Mbd2(-/-) ApoE(-/-)) mice (n=12, 24 eyes, experimental group) and MBD2 (wt) ApoE(-/-) mice (n=12, 24 eyes, control group) were fed on Western-type diet for 4 months. The mice were sacrificed, and total serum cholesterol levels were analyzed and Bruch's membrane (BM) of the eyes was removed for ultrastructural observation by transmission electron microscopy. Moreover, intercellular adhesion molecule 1 (ICAM-1) immunoreactivities were evaluated by fluorescence microscopy in sections of the eyes in both groups for further understanding the function mechanism of MBD2. There was no significant difference in the total serum cholesterol levels between control group and experimental group (P>0.05). Transmission electron microscopy revealed that AMD-like lesions, various vacuoles accumulated on BM, notable outer collagenous layer deposits and dilated basal infoldings of retinal pigment epithelium (RPE) were seen in both groups, and the BM in control group was significantly thickened as compared with experimental group (P<0.05). Fluorescence micrographs exhibited the expression of ICAM-1 in choroid was higher in control group than in experimental group. We are led to conclude that MBD2 gene knockout may lead to accumulation of more deposits on the BM and influence the pathogenesis of AMD via triggering endothelial activation and inflammatory response in choroid, improving microcirculation, and reducing lipid deposition so as to inhibit the development of AMD-like lesions. Our study helps to provide a new therapeutic approach for the clinical treatment of AMD.
Animals ; Apolipoproteins E ; genetics ; metabolism ; Bruch Membrane ; metabolism ; ultrastructure ; Cholesterol ; blood ; Choroid ; metabolism ; ultrastructure ; DNA-Binding Proteins ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Macular Degeneration ; blood ; genetics ; metabolism ; Male ; Mice ; Mice, Knockout ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Retinal Pigment Epithelium ; metabolism ; ultrastructure

OBJECTIVE: To observe the micromorphological characteristic of bacterial biofilm on mucosa of chronic rhinosinusitis (CRS). METHOD: Mucosa samples of middle turbinate were obtained from 4 patients of CRS during ESS. The size of each sample was about 4 mm x 4 mm. The samples were fixed in 4% paraformaldehyde for 24 hours, then fixed in 1% osmium tetroxide for 2 hours, graded dehydration with ethanol, dried with carbon dioxide and sputter coated with gold. The ultrastructure of these samples was observed by scanning electron microscope. RESULT: Bacterial biofilms were found on samples in all 4 patients. The biofilms were mainly formed on the surface of cilia. The bacterial flagella and cilia were intertwined. The biofilms could be found in a lot kinds of bacterial infections or mixed infections which were caused by multiple bacteria and fungi. CONCLUSION Bacterial biofilm could be formed on ciliated epithelia.
Bacterial Infections ; microbiology ; Biofilms ; Cilia ; microbiology ; ultrastructure ; Epithelium ; microbiology ; ultrastructure ; Humans ; Microscopy, Electron, Scanning ; Sinusitis ; microbiology

OBJECTIVE: To explore the postmortem changes of cornea thickness measured by ultrasonic pachymetry. METHODS: Eleven rabbits were randomly divided into two groups: one group with intact corneal epithelium and another group without intact corneal epithelium. In the later group, the corneal epithelium of the rabbit was scraped using mechanical elimination method. The corneal thickness was monitored continuously by ultrasonic pachymetry at several postmortem interval points in rabbits of the two groups. The changes of corneal thickness and postmortem interval were explored by relative regression analysis. RESULTS: The thickness of the cornea showed a strong non-linear correlation with the postmortem interval in the group with intact corneal epithelium. The group with intact corneal epithelium showed the correlation coefficient 0.922 and the group without intact corneal epithelium showed the correlation coefficient 0.822, respectively. CONCLUSION The corneal thickness measured by ultrasonic pachymetry shows a potential value for estimating early postmortem interval. The intact corneal epithelium is a crucial factor for the measurement of cornea thickness by ultrasonic pachymetry.
Animals ; Cornea/pathology* ; Corneal Topography/methods* ; Epithelium, Corneal/ultrastructure* ; Female ; Forensic Pathology/methods* ; Male ; Postmortem Changes ; Rabbits ; Regression Analysis ; Reproducibility of Results ; Time Factors ; Ultrasonography

OBJECTIVETo observe the expression of P-selectin in the penile vascular epithelial cells and the morphological changes in the ultrastructure of the penile cavernous tissues of smoking rats, and to explore the pathogenesis of smoking-induced erectile dysfunction.

METHODSFifty healthy Wistar rats were randomly divided into a normal control, a long-term heavy smoking group, a long-term light smoking, a short-term heavy smoking and a smoking cessation group. Their erectile function was tested by subcutaneous injection of apomorphine (APO), the P-selectin expression in the penile vascular epithelial cells detected by ELISA and the morphological changes in the ultrastructure of the penile cavernous tissues observed under the transmission electron microscope (TEM).

RESULTSThe levels of P-selectin were 10.78 +/- 1.71 ng/L, 62.62 +/- 5.95 ng/L, 40.06 +/- 3.97 ng/L, 41.37 +/- 4.06 ng/L and 22.80 +/- 3.15 ng/L respectively in the normal control, long-term heavy smoking, long-term light smoking, short-term heavy smoking and smoking cessation groups, with significant differences between the control group and the other four (P < 0.05). Electron microscopy showed abnormal arrangement of endothelia, penile cavernous sinuses and smooth muscle cells, disrupted continuity of endothelia, damaged ultrastructure of endothelial and smooth muscle cells in the penile cavernous tissue, and obvious proliferation and fibrosis of interstitial tissues in the smoking rats.

CONCLUSIONSmoking increases the P-selectin expression in the penile vascular epithelial cells and damages the ultrastructure of the penile cavernous tissue, which may be the main contributors to smoking-induced erectile dysfunction.

Animals ; Epithelium ; metabolism ; Male ; P-Selectin ; biosynthesis ; Penile Erection ; Penis ; blood supply ; metabolism ; ultrastructure ; Rats ; Rats, Wistar ; Smoking ; adverse effects

OBJECTIVETo investigate the impact of experimental varicocele (EV) on the ipsilateral testis in rats.

METHODSEV was induced by partial ligation of the left renal vein in male SD rats, the control rats subjected to sham operation, and the testes of the EV models and controls were extirpated 6, 12, and 18 weeks later. Johnson's score, ultrastructure of seminiferous tubules, intratesticular testosterone concentration (ITC) and germ cell apoptotic index (AI) of each left testis were evaluated.

RESULTSJohnson's scores were (6.92 +/- 0.52), (4.83 +/- 0.41) and (2.95 +/- 0.26), ITCswere (6.32 +/- 0.85), (5.17 +/- 0.76) and (4.11 +/- 0.69) and AIs were (5.32 +/- 1.23), (15.21 +/- 0.97) and (21.13 +/- 1.12) respectively in the 6 w , 12 w and 18 w EV groups, significantly lower than in the corresponding control groups, (9.56 +/- 0.35, 9.63 +/- 0.31, 9.39 +/- 0.46), (9.64 +/- 1.23, 9.38 +/- 0.69, 9.73 +/- 0.49) and (3.21 +/- 1.15, 3.43 +/- 1.21, 3.61 +/- 1. 15) (P < 0.05), the former two showing a gradual decline while the latter a significant elevation with the increasing duration of varicocele. The damage to the ultrastructure of seminiferous tubules was aggravated with the prolonging of varicocele.

CONCLUSIONEV can cause a progressive decline of ITC, dyszoospermia and increased AI of germ cells.

Animals ; Apoptosis ; Disease Models, Animal ; Infertility, Male ; Male ; Rats ; Rats, Sprague-Dawley ; Seminiferous Epithelium ; cytology ; ultrastructure ; Testis ; chemistry ; metabolism ; Testosterone ; metabolism ; Varicocele ; metabolism ; physiopathology

OBJECTIVETo study the correlation between cyclin E protein overexpression and centrosome amplification in oral squamous cell carcinoma (OSCC).

METHODSFormalin-fixed, paraffin-embedded tissues from 12 normal oral epithelium cases and 46 cases of OSCC were studied. Their centrosome status was analyzed by indirect immunofluorescence double staining with antibodies to centrosome protein gamma-tubulin and cytokeratin. The expression of cyclin E protein was studied by immunohistochemical methods. The correlation between cyclin E protein expression and centrosome amplification in OSCC was statistically analyzed by SPSS 12.0.

RESULTSThirty-seven of the 46 OSCC cases (80.4%) studied showed evidence of centrosome amplification, as signified by enlargement and/or increase in number of centrosomes, while normal oral epithelium possessed centromeres of normal size and number. Positive staining for cyclin E protein was observed in 30 of the 46 OSCC cases (65.2%), while all the normal oral epithelium cases were cyclin E protein-negative. The percentage of centrosome amplification in OSCC with positive cyclin E protein staining (90.0%, 27/30) was higher than that in OSCC with negative cyclin E protein staining (62.5%, 10/16) (chi(2) = 5.014, P < 0.05). Centrosome amplification showed positive correlation with cyclin E protein overexpression (r = 0.330, P < 0.05).

CONCLUSIONUp-regulation of cyclin E protein may represent one of the possible mechanisms for centrosome amplification in OSCC.

Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Centrosome ; pathology ; ultrastructure ; Cyclin E ; metabolism ; Epithelium ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Microscopy, Confocal ; Mouth Mucosa ; metabolism ; pathology ; Mouth Neoplasms ; metabolism ; pathology ; surgery ; Up-Regulation

The purpose of this study is to characterize and compare the ultrastructural changes occurring during the in vivo cultivation of corneal epithelium on amniotic membrane (AM) at several different time points. Corneal burn patients (n=7) with a corneal epithelial defect and severe limbal damage were selected. Initially, AM transplantation with limbal autograft was performed at the acute stage of corneal burn to reconstruct the damaged ocular surface. One to six (mean interval; 3.3+/-1.2) months later, the central part of AM containing an in vivo expanded corneal epithelium was excised and retransplanted in adjacent lesions. The excised epithelium with AM was examined by electron microscopy and immunohistochemical study. By electron microscopy, one and two months after expansion, cultivated epithelium on AM showed an undifferentiated epithelium and an incomplete basement membrane (BM). But, after three months, the cultivated epithelium began to differentiate into a multilayered epithelium with a continuous BM with increased hemidesmosomes. These findings were further confirmed by immunohistochemical study, that cytokeratin K3 was expressed in the cultivated corneal epithelium and newly formed BM was partially positive of collagen IV at three months. At least 3 months may be needed for the proliferation and differentiation of in vivo cultivated corneal epithelium on AM.
Stem Cells/cytology ; Stem Cell Transplantation/*methods ; Middle Aged ; Microscopy, Electron ; Male ; Keratin-3/biosynthesis ; Immunohistochemistry ; Humans ; Epithelium, Corneal/cytology/*metabolism/*pathology/*transplantation ; Corneal Diseases/*therapy ; Burns/*surgery/therapy ; Biological Dressings ; Amnion/*ultrastructure ; Adult