Degeneration Of Epithelial Cells Research Articles

Lamb model of respiratory syncytial virus-associated lung disease: insights to pathogenesis and novel treatments.

Preterm birth is a risk factor for respiratory syncytial virus (RSV) bronchiolitis and hospitalization. The pathogenesis underlying this is not fully understood, and in vivo studies are needed to better clarify essential cellular features and molecular mechanisms. Such studies include analysis of lung tissue from affected human infants and various animal models. The preterm and newborn lamb lung has developmental, structural, cellular, physiologic, and immunologic features similar to that of human infants. Also, the lamb lung is susceptible to various strains of RSV that infect infants and cause similar bronchiolar lesions. Studies in lambs suggest that viral replication in airways (especially bronchioles) is extensive by 4 days after infection, along with bronchiolitis characterized by degeneration and necrosis of epithelial cells, syncytial cell formation, neutrophil infiltration, epithelial cell hypertrophy and hyperplasia, and innate and adaptive immune responses. RSV bronchiolitis greatly affects airflow and gaseous exchange. RSV disease severity is increased in preterm lambs compared with full-term lambs; similar to human infants. The lamb is conducive to experimental assessment of novel, mechanistic therapeutic interventions such as delivery of vascular endothelial growth factor and enhancement of airway epithelial oxidative responses, Club (Clara) cell protein 10, and synthesized compounds such as nanobodies. In contrast, exposure of the fetal ovine lung in vivo to ethanol, a risk factor for preterm birth, reduces pulmonary alveolar development and surfactant protein A expression. Because the formalin-inactivated RSV vaccination enhances some inflammatory responses to RSV infection in lambs, this model has the potential to assess mechanisms of formalin-inactivated RSV enhanced disease as well as newly developed vaccines.

The effect of Qi'ao Decoction on ovalbumin induced and lipopolysaccharide enhanced severe asthma mice and its mechanism

AimTo evaluate the effect of Qi'ao Deocoction (QAD) on the inflammation and hyperresponsiveness of asthma mice. Methods120 Balb/C mice were randomly divided into six groups: normal group, model group, dexamethasone group, high dose QAD group, medium dose QAD group and low dose QAD group. The asthma model was reproduced in Balb/C mice sensitized by ovalbumin, challenged by OVA and LPS. The mice of the normal group were sensitized, chanllenged and intranasally instilled by PBS. On day 28-34, 6.7, 13.4 and 26.8 g·kg−1 Qi'ao Decoction were administrated; 0.002 4 g·kg−1 dexamethasone solution was given to the dexamethasone group; normal and model groups were given the same amount of normal saline. Bronchoalveolar lavage fluid, airway hyperresponsiveness, lung histopathology and cytokines were then collected and analyzed. ResultsCompared with normal group, total cellular score, the number of macrophages, lymphocytes, eosinophils and neutrophils of model group significantly increased (P < 0.01). Compared with model group, the administration of dexamethasone induced a significant decrease in eosinophils and neutrophils (P < 0.05, P < 0.01). The number of eosinophils, which plays an important role in airway inflammatory reaction of asthma, of the three QAD groups all decreased (P < 0.01). RL before and after Ach (5 mg·mL−1) stimulation in the model group both overtook that in the normal group (P < 0.01). Compared with model group, dexamethasone group, high dose QAD group, medium dose QAD group and low dose QAD group groups all had significantly lower RL before and after Ach stimulation (P < 0.01). Normal pulmonary histopathology was found in the normal group. In the model group, mice exhibited marked increases in inflammatory cell infiltration, mostly including neutrophils and macrophages, perivascular inflammation and thickened alveolus wall (P < 0.01). Dexamethasone application mitigated inflammation around the bronchi (P < 0.05). These histopathological changes were ameliorated in the three decoction groups (P < 0.01, P < 0.05). In addition, alveolus and airway wall lesions of medium dose QAD group and high dose QAD group were reduced, the number of inflammatory cells infiltrated around the walls decreased, no clear degeneration of bronchial epithelial cells was found, and exudates in bronchi declined in different degrees. Compared with normal group, IFN-γ and IL-12 of model group significantly decreased, while IL-4 increased, showing statistic difference (P < 0.05). Compared with model group, IFN-γ and IL-12 level of dexamethasone group went up too, but IL-4 declined (P < 0.05). The level of IFN-γ of medium dose QAD group and high dose QAD group both increased; IL-4 and IL-12 of medium dose group were found significant differences (P < 0.05); but none of the cytokines of low dose QAD group showed statistical significance (P > 0.05). ConclusionQAD can significantly inhibit airway inflammation and airway hyperresponsiveness of mice with severe asthma induced by ovalumin and lipopolysaccharide, adjust the balance of cytokines, and improve lung histopathological condition. So, it exhibits great effect on severe asthma.

Nephritis Associated with a S1 Variant Brazilian Isolate of Infectious Bronchitis Virus and Vaccine Protection Test in Experimentally Infected Chickens

4 Abstract: Infectious bronchitis virus (IBV) induces a significant negative impact on poultry production worldwide, specially due to the continuous emergence of viral variants. This virus causes damage to th e respiratory tract, and depending on the virus strain, affects and damages the urogenital system. The objective of this study is to characterize the pathotype and the cross-immunity with regard to Massachusetts vaccine strain (H120) of a Brazilian IBV field isolate (IBVPR-05) previously S1 genotyped as a variant. The pathogenicity test was conducted on two experimental groups of specific pathogen free chickens; one was vaccinated with attenuated Massachusetts H120 strain and the other remained non-vaccinated. Three weeks after vaccination, both groups were challenged with IBRPR-05. The tracheal ciliostasis, and the viral load, histopathology and immunohistochemistry in trachea and kidney samples were evaluated. The viral loads, measured by quantitative real time RT-PCR, were higher in kidney than in trachea, and the most prominent histopathological changes were found in the kidneys. The renal lesions were characterized by the presence of nephritis with intense inflammation, tubular epithelial cell degeneration and necrosis. The H120 vaccine induced a partial protection against the infection of trachea and kidney tissues by this variant isolate. Thus, the Brazilian variant isolate IBVPR-05 was characterized in this study as a nephropathogenic pathotype and as a pr otectotype differing from Massachusetts vaccine strain of IBV. This indicates the importance to determine these biologic characteristics of other Brazilian variant IBV isolates, in order to implement more effective control measures of IBV infection in this country.

Involvement of autoimmunity to REG, a regeneration factor, in patients with primary Sjögren's syndrome

The regenerating gene (Reg) was isolated originally as a gene specifically over-expressed in regenerating pancreatic islets and constitute a growth factor family. Reg gene product (Reg) is important in the pathophysiology of various human inflammatory diseases. Recently, the possible involvement of human REG in the regeneration of salivary ductal epithelial cells of patients with primary Sjögren's syndrome (SS) was reported. However, the expression of the REG family genes in minor salivary glands (MSG) and the occurrence of anti-REG Iα autoantibodies in SS patients were obscured. In this study, we examined the expression of REG family genes in the MSG of SS and screened anti-REG Iα autoantibodies in SS. The mRNA levels of REG family genes in MSG were quantified using real-time reverse transcription-polymerase chain reaction (RT-PCR) and REG Iα expression in the MSG was analysed by immunohistochemistry. The mRNA level of REG Iα in the MSG of SS patients was significantly higher than that of control. REG Iα protein was expressed highly in SS ductal epithelial cells. Anti-REG Iα autoantibodies in the sera were found in 11% of SS. All the MSG in the anti-REG Iα autoantibody-positive group showed REG Iα expression, whereas only 40% showed REG Iα expression in the anti-REG Iα autoantibody-negative group. The anti-REG Iα autoantibody-positive group showed significantly lower saliva secretion and a higher ratio of grade 4 (by Rubin-Holt) in sialography. These data suggest strongly that autoimmunity to REG Iα might play a role in the degeneration of MSG ductal epithelial cells in primary SS.

Epigallocatechin-gallate (EGCG) regulates autophagy in human retinal pigment epithelial cells: A potential role for reducing UVB light-induced retinal damage

Autophagy is an intracellular catabolic process involved in protein and organelle degradation via the lysosomal pathway that has been linked in the pathogenesis of age-related macular degeneration (AMD). UVB irradiation-mediated degeneration of the macular retinal pigment epithelial (RPE) cells is an important hallmark of AMD, which is along with the change in RPE autophagy. Thus, pharmacological manipulation of RPE autophagy may offer an alternative therapeutic target in AMD. Here, we found that epigallocatechin-3-gallate (EGCG), a polyphenolic compound from green tea, plays a regulatory role in UVB irradiation-induced autophagy in RPE cells. UVB irradiation results in a marked increase in the amount of LC3-II protein in a dose-dependent manner. EGCG administration leads to a significant reduction in the formation of LC3-II and autophagosomes. mTOR signaling activation is required for EGCG-induced LC3-II formation, as evidenced by the fact that EGCG-induced LC3-II formation is significantly impaired by rapamycin administration. Moreover, EGCG significantly alleviates the toxic effects of UVB irradiation on RPE cells in an autophagy-dependent manner. Collectively, our study reveals a novel role of EGCG in RPE autophagy. EGCG may be exploited as a potential therapeutic reagent for the treatment of pathological conditions associated with abnormal autophagy.

Autophagy Activation Clears ELAVL1/HuR-Mediated Accumulation of SQSTM1/p62 during Proteasomal Inhibition in Human Retinal Pigment Epithelial Cells

Age-related macular degeneration (AMD) is the most common reason of visual impairment in the elderly in the Western countries. The degeneration of retinal pigment epithelial cells (RPE) causes secondarily adverse effects on neural retina leading to visual loss. The aging characteristics of the RPE involve lysosomal accumulation of lipofuscin and extracellular protein aggregates called “drusen”. Molecular mechanisms behind protein aggregations are weakly understood. There is intriguing evidence suggesting that protein SQSTM1/p62, together with autophagy, has a role in the pathology of different degenerative diseases. It appears that SQSTM1/p62 is a connecting link between autophagy and proteasome mediated proteolysis, and expressed strongly under the exposure to various oxidative stimuli and proteasomal inhibition. ELAVL1/HuR protein is a post-transcriptional factor, which acts mainly as a positive regulator of gene expression by binding to specific mRNAs whose corresponding proteins are fundamental for key cellular functions. We here show that, under proteasomal inhibitor MG-132, ELAVL1/HuR is up-regulated at both mRNA and protein levels, and that this protein binds and post-transcriptionally regulates SQSTM1/p62 mRNA in ARPE-19 cell line. Furthermore, we observed that proteasomal inhibition caused accumulation of SQSTM1/p62 bound irreversibly to perinuclear protein aggregates. The addition of the AMPK activator AICAR was pro-survival and promoted cleansing by autophagy of the former complex, but not of the ELAVL1/HuR accumulation, indeed suggesting that SQSTM1/p62 is decreased through autophagy-mediated degradation, while ELAVL1/HuR through the proteasomal pathway. Interestingly, when compared to human controls, AMD donor samples show strong SQSTM1/p62 rather than ELAVL1/HuR accumulation in the drusen rich macular area suggesting impaired autophagy in the pathology of AMD.

Antioxidant Activity of Coated Probiotic Lactobacillus casei on Chromium(VI) Induced Oxidative Stress in Rats

Experimental study was conducted to evaluate the antioxidant potential of sodium alginate coated probiotic Lactobacillus casei against chromium (VI)-induced oxidative stress in albino rats. Administration of chromium (Cr) VI significantly (p < 0.05) increased the peroxidation markers such as malondialdehyde and protein carbonyls along with significant (p < 0.05) decrease in antioxidant markers such as super oxide dismutase and reduced glutathione in liver and kidney homogenates. Chromium also caused a significant (p < 0.05) alteration in hepatic and renal functional markers in serum viz., total protein, alanine transaminase, blood urea nitrogen and creatinine. Prominent pathological changes observed in liver were vascular congestion, degenerative changes, dilatation of sinusoids and mononuclear cell infiltration, while the kidney showed degeneration of tubular epithelial cells, cystic dilatation of tubules and hyaline casts. Co-administration of coated probiotic L. casei strain 17 reversed the Cr-induced changes.

The ultra-early protective effect of ulinastatin on rabbit acute lung injury induced by paraquat

ObjectiveTo study ultra-early pathophysiological changes of rabbit acute lung injury (ALI) caused by paraquat (PQ) and discuss the ultra-early protective effect of ulinastatin on rabbit ALI due to PQ.Methods30 New Zealand white rabbits were randomly divided into a control group, a paraquat group and an ulinastatin intervention group with 10 rabbits in each group. For paraquat group and intervention group a single dose of paraquat (35mg/kg) was injected intraperitoneally to establish rabbit models of ALI. The control group was injected an equal volume of saline. The intervention group was treated with 100Ku/kg ulinastatin immediately after the establishment of the ALI model. The respective experimental groups underwent 320-slice CT perfusion scan of pleural at 2h, 4h and 6h time point after modeling to get CTP (CT Perfusion) images and related parameters. 2mL blood was collected in the marginal ear vein to determine the mass concentration of the vascular endothelial growth factor (VEGF). The animals were killed by air embolism after 6h and lung tissue was taken for pathology observation.ResultsThe reginal blood flow (rBF) and reginal blood volume (rBV) of paraquat group at 2,4,6 h time point were significantly (P <0.05) lower than those of control group. The intervention group rBF and rBV at 2, 4 and 6 h time points were significantly higher (P <0.05) compared to paraquat group. The permeability surface (rPS) and VEGF mass concentration of paraquat group at 2,4,6 h time point were significantly higher than the control group (P <0.05), and the intervention group rPS and VEGF mass concentrations at 2,4,6h time point were significantly lower (P <0.05) than those of paraquat group. Pathological detection indicators of paraquat group (congestive capillary percentage, the number of red blood cells outside of capillaries, percentage of capillaries with basement membrane damage) were significantly higher (P <0.05) at 6h time point compared with the control group, while significantly lower (P <0.05) in intervention group than in paraquat groups. Pathological observation under light microscope showed in paraquat group obvious inflammatory cell infiltration, alveolar epithelial cell hyperplasia, widened alveolar septum, visible focal hemorrhage, visible acute and chronic inflammatory cell infiltration in bronchioles cavity; under electron microscopy alveolar epithelial cell degeneration and necrosis, vascular welling of the endothelial cells, basement membrane rupture, a lot of exudates in alveolar space. In the intervention group, the above the symptoms were mitigated.ConclusionIn the ultra-early stage of rabbit ALI induced by PQ, pulmonary vascular endothelial cell is damaged and serum VEGF mass concentration and pulmonary vascular permeability increase. Early ulinastatin intervention can reduce serum VEGF level and PQ-induced vascular permeability amplitude, indicating that ulinastatin has a protective effect on pulmonary vascular endothelial cells.

Preclinical Evaluation of the Toxicological Effects of a Novel Constrained Ethyl Modified Antisense Compound Targeting Signal Transducer and Activator of Transcription 3 in Mice and Cynomolgus Monkeys

ISIS 481464 is a constrained ethyl (cEt) modified phosphorothioate antisense oligonucleotide (ASO) targeting signal transducer and activator of transcription 3 (STAT3) studied in mice and monkey to support oncology clinical trials. Six-week toxicology studies were performed in mice and cynomolgus monkey (up to 70 and 30 mg/kg/week respectively). Reduction in STAT3 protein up to 90% of control was observed in monkey. Cynomolgus monkey was considered the most relevant species to human with respect to pharmacokinetic properties, but mice are useful in their relative sensitivity to the potential proinflammatory and hepatic effects of oligonucleotides. In monkeys, there was no impact on organ function at doses up to 30 mg/kg/week for 6 weeks. Minimal to slight proximal tubular epithelial cell degeneration and regeneration within the kidney was observed, which had no impact on renal function and showed reversibility at the end of the treatment-free period. Additionally, mild and transient activated partial thromboplastin time elevations and mild increases in complement Bb were observed at the higher doses by intravenous dosing only. In mice, the alterations at 70 mg/kg/week included spleen weight increase up to 1.4-fold relative to control, increases in alanine aminotransferase and aspartate aminotransferase up to 1.8-fold over control, interleukin-10 increases up to 3.7-fold, and monocyte chemoattractant protein-1 increase up to 1.9-fold over control. No significant clinical pathology or histopathology changes were seen in mice at 20 mg/kg/week or less. The toxicity profile of ISIS 481464 is consistent with effects observed with phosphorothioate ASOs containing 2'-O-methoxyethylribose modifications instead of cEt.

TOXICOLOGICAL STUDIES ON ISEPAMICIN (HAPA-B)

Acute toxicity of isepamicin (HAPA-B), a new aminoglycoside antibiotic, in mice, rats and dogs was examined in comparison with amikacin (AMK) and gentamicin (GM). Intravenous LD50 values of HAPA-B were 234 mg/kg in male and 236 mg/kg in female for mice, 489 mg/kg in male and 476 mg/kg in female for rats and 720-864 mg/kg for dogs. Those of AMK were 183 mg/kg in male and 181 mg/kg in female for mice, 420 mg/kg in male and 417 mg/kg in female for rats. Those of GM were 50 mg/kg in male and 47 mg/kg in female for mice, 119 mg/kg in male and 124 mg/kg in female for rats. Intraperitoneal LD50 values of HAPA-B were 2,244 mg/kg in male and 2,272 mg/kg in female for mice, 1,664 mg/kg in male and 1,591 mg/kg in female for rats. Intramuscular LD50 values of HAPA-B were 2,508 mg/kg in male and 2,632 mg/kg in female for mice, 2,088 mg/kg in male and 2,111 mg/kg in female for rats and greater than 1,800 mg/kg for dogs. Those of AMK were 1,247 mg/kg in male and 1,334 mg/kg in female for mice, 2,324 mg/kg in male and 2,244 mg/kg in female for rats. Those of GM were 359 mg/kg in male and 360 mg/kg in female for mice, 559 mg/kg in male and 557 mg/kg in female for rats. Subcutaneous LD50 values of HAPA-B were 3,321 mg/kg in male and 3,320 mg/kg in female for mice, 3,451 mg/kg in male and 3,392 mg/kg in female for rats. Oral LD50 values of HAPA-B were more than 5,000 mg/kg in mice and rats. Ataxia, acratia, dyspnea and convulsions were observed following administration by all routes, except for oral route, of all drugs in mice, rats and dogs. The cause of early death was due to respiratory paralysis which is the typical acute toxic sign of aminoglycoside antibiotics, and that of late death was due to renal injuries. BUN and creatinine values of surviving dogs after day 14 increased after administration by either intravenous or intramuscular routes. Disintegration, necrosis and calcification of epithelial cells of the proximal convoluted tubuli were observed in rats which died during the course of the study, and atrophy, dilatation and eosinophilic degeneration in epithelial cells of the proximal convoluted tubuli and thickening of Bowman's capsule were observed in surviving dogs.

TOXICOLOGICAL STUDIES ON ISEPAMICIN (HAPA-B)

Subacute toxicity in the rat of isepamicin (HAPA-B), a new aminoglycoside antibiotic, was examined in comparison with amikacin (AMK). Daily doses of 12.5, 25, 100 and 300 mg/kg of HAPA-B or 25 and 100 mg/kg of AMK were injected intramuscularly for 28 days. No animal died and there were no changes in general symptoms except for hemorrhage at injection sites of both drugs. Decreases in body weight gain and food consumption were observed in males and females in the HAPA-B 300 mg/kg dose group. Water consumption was increased in males in the HAPA-B 300 mg/kg and AMK 100 mg/kg dose groups. Elevations of serum creatinine and/or BUN in males and females were occasionally observed in the HAPA-B 300 mg/kg and AMK 100 mg/kg dose groups, and elevation of GOT, which seemed to be due to muscle injuries, was occasionally observed in a no dose-dependent manner with either drug. Increases in kidney and caecum weights of males and females in the HAPA-B 100, 300 mg/kg and AMK 25, 100 mg/kg dose groups and increases in adrenal relative-weights in males and females in the HAPA-B 300 mg/kg dose group were observed. At necropsy, discoloration and enlargement of the kidney were observed in a dose-dependent manner at or above 100 mg/kg with either drug. Histopathological findings indicated degeneration and necrosis in epithelial cells of the proximal convolute tubuli and thickening of basement membrane. Electron microscopic findings indicated an increase in number of large lysosomes containing myeloid bodies in the epithelial cells of the proximal convolute tubuli with both drugs. The above results suggest that HAPA-B has renal toxicity like other aminoglycoside antibiotics but its toxicity was lower than AMK. The non-toxic dose of HAPA-B in this test was 12.5 mg/kg.

The role of neuropeptide Y and aquaporin 4 in the pathogenesis of intestinal dysfunction caused by traumatic brain injury

BackgroundAlthough the exact incidence is unknown, traumatic brain injury (TBI) can lead to intestinal dysfunction. It has important influence on the early nutrition and prognosis of TBI patients. Experiments were designed to study the roles of neuropeptide Y (NPY) and aquaporin 4 (AQP4) in the pathogenesis of intestinal dysfunction caused by TBI and to find some new solutions for the treatment of intestinal dysfunction after TBI. MethodsForty adult male Wistar rats were randomly divided into control, mild trauma, moderate trauma, and severe trauma groups. TBI was induced by Feeney's impact method. Control animals were sham operated but not subjected to the impact test. All rats were killed 24 h after surgery. Blood samples were obtained from the abdominal aorta for enzyme-linked immunosorbent assay measurement of NPY concentrations. Jejunum segments 15 cm distal to the Treitz ligament were taken for analysis of NPY and AQP4 expression by polymerase chain reaction, Western blot, and immunohistochemistry. Pathologic changes in intestinal cell structure and ultrastructure were studied by light microscopy and transmission electron microscopy. ResultsThe specimens from different groups showed different degrees of structural changes, ranging from swelling and degeneration of villous epithelial cells to extensive denudation and collapse of the villi. The more severe the trauma, the more serious the degree of intestinal mucosal injury. Intestinal smooth muscle also showed varying degrees of edema and structural disorder. Electron microscopy showed that intestinal mitochondria had varying degrees of swelling and the structure of mitochondrial crista was disordered and even fractured. Plasma concentrations of NPY and jejunal gene and protein expressions of NPY and AQP4 increased significantly following TBI (P < 0.05), with greater increases at higher levels of injury. Moreover, there were positive correlations between NPY and AQP4 (P < 0.05). ConclusionsIncreasing grades of TBI caused increasing degrees of intestinal ischemia and edema, and thus caused increasingly severe intestinal dysfunction. AQP4 and NPY may be involved in the pathogenesis of intestinal dysfunction after TBI. Increased NPY levels may be responsible for intestinal ischemia and hypoxia, and AQP4 may play an important role in intestinal edema. Increased NPY levels may be one of the main causes for the increase in AQP4 after TBI.

Histological study on conjunctival and corneal reactions in rabbits induced by chronic topical application of latanoprost and travoprost

Introduction: Chronic topical glaucoma therapy was reported to cause deleterious changes tothe ocular surface epithelial layer. Aim of the work: The aim of this study was to compare the histological changes in the cornea after chronic exposure to latanoprost preserved with 0.02% benzalkonium chloride(BAK) eye drops, travoprost preserved with sofZia eye drops and preservative-free artificial tears. Materials and methods Fifteen white rabbits were randomized into three groups (five animals each). They received once-daily topical application of one of the three treatments for 30 days. The first group (the control group) received preservative-free artificial tears (Refresh Plus). The second group received travoprost preserved with SofZia (Travatan Z).The third group received latanoprost preserved with 0.02% BAK (Xalatan). Enucleation was performed at the end of the experiment. Corneal samples were processed for light and transmission electron microscopic studies and conjunctival samples for light microscopic study. The mean epithelial height of the corneal epithelium, the mean number of goblet cells in the conjuctival epithelium and the mean area% of PAS-positive goblet cells were measured using an image analyzer. These results were statistically analysed using analysis of variance and the t-test. Results: Latanoprost eye drops preserved in BAK produced toxic changes in the form of degeneration and desquamation of the superficial epithelial cells in the cornea, separation of Descemet’s membrane and degeneration of endothelial cells, in addition to decreased number of goblet cells in the conjunctiva. Travoprost eyedrops preserved in sofZia were safer and produced slight changes on the rabbit’s ocular surface compared with latanoprost eye drops preserved in BAK. Conclusion It was concluded that antiglaucomatous drugs preserved with sofZia produced less corneal and conjunctival changes than those preserved with BAK.

Derivation, characterization and retinal differentiation of induced pluripotent stem cells

Millions of people world over suffer visual disability due to retinal dystrophies which can be age-related or a genetic disorder resulting in gradual degeneration of the retinal pigmented epithelial (RPE) cells and photoreceptors. Therefore, cell replacement therapy offers a great promise in treating such diseases. Since the adult retina does not harbour any stem cells, alternative stem cell sources like the embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer a great promise for generating different cell types of the retina. Here, we report the derivation of four iPSC lines from mouse embryonic fibroblasts (MEFs) using a cocktail of recombinant retroviruses carrying the genes for Oct4, Sox2, Klf4 and cMyc. The iPS clone MEF-4F3 was further characterized for stemness marker expression and stable reprogramming by immunocytochemistry, FACS and RT-PCR analysis. Methylation analysis of the nanog promoter confirmed the reprogrammed epigenetic state. Pluripotency was confirmed by embryoid body (EB) formation and lineage-specific marker expression. Also, upon retinal differentiation, patches of pigmented cells with typical cobble-stone phenotype similar to RPE cells are generated within 6 weeks and they expressed ZO-1 (tight junction protein), RPE65 and bestrophin (mature RPE markers) and showed phagocytic activity by the uptake of fluorescent latex beads.

Magnifying Endoscopy With Narrow Band Imaging and Endoscopic Ultrasonography for Assessing Eosinophilic Esophagitis

A 59-year-old man presented with epigastric pain during the past 4 weeks. Because his symptom was aggravated with ingestion of food, he complained of poor oral intake and weight loss of 4 kg. His medical history included bronchial asthma which had been diagnosed 4 months before, but he took medicine irregularly. Physical examination was unremarkable. Laboratory studies showed peripheral eosinophilia (920/µL, 14.5% of the circulating leukocytes), and an elevated immunoglobulin E level (895.2 IU/mL). Computed tomography of abdomen demonstrated diffuse thickening of the distal esophageal wall (Fig. 1). On upper endoscopy, the distal esophagus was edematous and pale with adherent pin point whitish exudates (Fig. 2A). Magnifying endoscopy with narrow band imaging demonstrated multiple white micronodules on the mucosal surface (Fig. 2B). Endoscopic ultrasonography (EUS) showed circumferential, mainly thickening of the deep mucosal/submucosal and inner circular muscle layers (Fig. 2C). We took biopsy specimens from proximal, middle and distal esophagus, and only biopsy from distal esophagus showed > 20 eosinophils per high-power field infiltrating the esophageal mucosa (Fig. 3A). Before the histological diagnosis of eosinophilic esophagitis, we tried proton pump inhibitor twice daily for him. However, proton pump inhibitor did not do any good, and finally the patient was diagnosed with eosinophilic esophagitis. Skin prick tests for inhalant and food allergens showed negative results. Treatment was initiated with orally administered fluticasone 500 µg 3 times a day. Epigastric pain improved rapidly. At 8 weeks after treatment, endoscopic features improved (Fig. 2D and 2E), and EUS showed decreased thickness of each layer of the esophagus (Fig. 2F). Histologic remission was also achieved (Fig. 3B). After 3 months of treatment with fluticasone, he stopped the medicine. At present, 6 months have passed without recurrence of symptom. Figure 1 Computed tomography of abdomen demonstrates diffuse thickening of the distal esophageal wall. Figure 2 Endoscopic findings (A-C, before treatment; D-F, after treatment). (A) Upper endoscopy shows edematous and pale esophagus with adherent pin point whitish exudates. (B) On magnifying endoscopy with narrow band imaging, there are multiple white micronodules ... Figure 3 Histologic findings. (A) Biopsy specimen from the distal esophagus shows greater than 20 eosinophils per high power field infiltrating the esophageal mucosa associated with epithelial cell degeneration. And eosinophilic microabscesses (white arrow) are ... The roles of magnifying endoscopy with narrow band imaging and EUS for assessing eosinophilic esophagitis are not clear yet.1-3 However, looking at this case, those could be useful tools, especially when typical endoscopic features such as concentric rings and linear furrows are not seen by conventional endoscopy.

Histopathological Studies on Stunting Syndrome in Broilers, Lahore, Pakistan

Runting stunting syndrome (RSS) is a multifactorial disease with many names and faces that had caused considerable economic losses to poultry through reduced uniformity, reduced livability, decreased body weights, elevated feed conversions, and many secondary diseases. The aim of the current study was to evaluate the effect of stunting syndrome on histopathology in chicks (n = 120) of different ages collected from nine different farms. Grouping was done on the basis of age (G1 = 1–10 days, G2 = 11–20 days, G3 = 21–30 days, and G4 = 31–40 days) including both stunted and normal chicks. Histopathological findings were the intestinal lesions (29%), including degeneration of villi, crypts, epithelial cells and lamina propria. Pancreatic histopathological lesions (16.65%) included the fibrosis, vacuolation, and degeneration of acinar cells. Degeneration of follicles and epithelial cells, of bursa of fabricius (43%) and dilation of glandular cells of proventriculus including lymphocytes infiltration (5.6%) were other histopathological findings. All these changes may interfere with normal digestive processes and normal body functioning resulting in poor weight gain and retarded growth or stunting of chicks.

Vanadium Toxicity in Mice

Because precise information as to the toxicity of vanadium is required for practical use of vanadium compounds as antidiabetic drugs, we examined vanadium toxicity in mice fed normal diet or high-fat diet (C57BL/6N, male, 7 weeks) by oral administration of ammonium metavanadate (AMV) with a maximum dose of 20 mgV/kg/day. Marked lipid accumulation in hepatocytes, renal epithelial cells, and mucosal epithelial cells of the small and large intestines and severe degeneration, necrosis, and loss of mucosal epithelial cells in the small intestine were observed. These pathological changes were more severe in mice fed high-fat diet than mice fed normal diet, and the intensity of the changes increased with increase in the administered dose of AMV. By electron microscopy, the number and size of lipid droplets in hepatocytes were increased. In the small intestine, a TUNEL assay showed a decreased number of positive cells, and positive cells for acrolein immunohistochemistry were observed specifically in the mucosal epithelial cells indicating degeneration and necrosis in the AMV-treated group, suggesting that a possible factor responsible for cell necrosis in the small intestine could be oxidative stress. In conclusion, AMV may impair cellular lipid metabolism, resulting in lipid accumulation, and induce mucosal epithelial cell necrosis in the small intestine.

Advanced Glycation End Products and Ultrastructural Changes in Corneas of Long-term Streptozotocin-Induced Diabetic Monkeys

The purpose of this study was to investigate the ultrastructural corneal changes of chronic diabetic monkeys and explore the relationship between advanced glycation end products and ultrastructural changes in diabetic corneas. A total of 8 cynomolgus monkeys were used in this experiment. Four monkeys were induced into insulin-dependent diabetes mellitus for 4 years. Four age-matched healthy monkeys were used as the controls. Ultrathin sections obtained from the corneas were examined by transmission electron microscopy. Advanced glycation end product immunoreactivity was observed in the epithelial cells, epithelial basement membrane, and stromal keratocytes of diabetic corneas, whereas advanced glycation end product immunoreactivity was not found in the corresponding area in normal corneas. Abnormal collagen fibril bundles of variable thickness were identified in corneal stroma in all diabetic monkeys. Epithelial and endothelial cell degeneration was also observed in 1 diabetic monkey. Abnormal aggregates of collagen fibrils in stromal matrix were common among long-term diabetic monkeys, and the formation of the abnormal collagen fibril aggregates might result from excessive nonenzymatic glycosylation.

Toxicity of ZnO nanoparticles in healthy adult mice

In this study, we extended previous work to evaluate the oral toxicity of ZnO nanoparticles and their possible effects on different serum-elements and sexual hormones in the mouse. The histopathological changes have also been examined. Significant recorded increases in alanine aminotransferase and aspartate aminotransferase activity in all mice exposed to ZnO nanoparticles suggest that these nanoparticles can cause hepatic injury. Hepatocyte necrosis and other pathological observations also confirmed liver damage. Moreover, Glomeruli segmentation, hydropic degeneration in epithelial cells, necrosis of epithelial cells in tubules and swelling in epithelial cells of proximal tubules were found in all kidney tissues, which demonstrated that ZnO nanoparticles have severe toxicological effects on kidney. Serous inflammation, severe hyperemia in alveoli, and edema were observed as pathological findings in the lung which suggest that the lung is the third target tissue of the ZnO nanoparticles.

Clinical analysis on bile reflux gastritis in children

Objective To compare the clinical symptoms,endoscopic lesions,pathological histological changes,and H.pylori infection between children with bile reflux gastritis (BRG) and non BRG.Methods 287 cases of children with upper gastrointestinal symptoms were diagnosed with gastritis by gastroscopy.They included 58 cases of BRG and 229 cases of non-BRG.The clinical symptoms,endoscopic lesions,pathological histological changes,and H.pylori infection were compared between two groups,respectively.Results Between BRG and non-BRG groups,there werent statistically significant difference in the main clinical symptoms including abdominal pain,anorexia,nausea,and early satiety.Incidence of abdominal distension,vomiting,and hiccups was[84.5％ (49/58) vs 66.8％ (153/229),32.8％ (19/58) vs 17.9％ (41/229),and 29.3％ (17/58) vs 16.6％ (38/229)],respectively,with statistical significance (x2 =6.93,6.18,4.83,P ＜0.05).There weren(t) statistically significant difference in endoscopic lesions congestion,edema,and mucosal plaques between two groups.Incidence of endoscopic gastric erosions and bleeding of two groups was [37.9％ (22/58) vs 22.7％ (52/229),20.7％ (12/58) vs 10.0％ (23/ 229)],respectively; and BRG group was statistically higher than non-BRG group(x2 =5.60,4.90,P ＜ 0.05).For micronodular change,there were 13 cases (22.4％) in BRG group and 86 cases (37.6％) in Non-BRG group,and Non-BRG group was statistically higher than BRG group(x2 =4.69,P ＜ 0.05).For histopathological changes including epithelial cell degeneration,hyperplasia and lamina propria lymphocyte and neutrophil infiltration,there wasnt statistically significant difference between BRG and non-BRG groups.Incidence about pit extension and mucosal vascular expansion of two groups was [10.34％ (6/58) vs 3.1％ (7/229) and 31.0 ％ (18/58) vs 18.3 ％ (42/229)],respectively ; and BRG group was significantly higher than non-BRG group(x2 =5.97,4.51,P ＜ 0.05).There were 7 cases (12.1％) in BRG group and 55 cases (24.0％) about lamina propria lymphoid follicles,and Non-BRG group was significantly higher than BRG group (x2 =3.90,P ＜0.05).H.pylori positive rates were[22.41％ (13/58),and 40.17％ (92/ 229)],respectively.BRG group and non-BRG group were found statistically significant (x2 =6.29,P ＜0.05).Conclusions BRG children with the characteristic symptoms were abdominal distension,vomiting,and hiccup inverse.Endoscopic characteristic performance was gastric erosions,bleeding spots in antral mucosa changes apparently.Mucosal vascular expansion and pit extension may prompt gastric in children with duodenogastric reflux.H.pylori positive rates of those gastric in children with bile reflux were relatively low.The clinical features could help early diagnosis BRG and guide clinical treatment to improve the cure rate. Key words: Bile reflux; Gastritis/etiology; Child