Effects of erlotinib on histopathology and ultrastructure of corneal epithelium in mice / 眼科新进展

Objective To investigate the histopathological and ultrastructural changes of corneal epithelium induced by erlotinib in mice.Methods Totally 30 6-8 weeks old male BALB/c mice were divided into three groups:Control group (n =12),experimental group (n =12),another 6 mice did nothing as the blank control.Experimental group used erlotinib eye drops and control group used PBS in both eyes,four times per day.At 1 day,7 days and 14 days after the intervention,corneal fluorescence staining (FL) was observed by slit lamp and graded.On the fourteenth day after the intervention,the eye balls of mice were taken,and the histopathological and ultrastructural changes of corneal epithelium and epithelial cells were observed by optical microscope and electron microscope,respectively.And protein of cornea was measured by Western Blot.Results Before the intervention,there was no significant difference in FL scores between the experimental group and control group (P ＞ 0.05).At 1 day,7 days and 14 days,FL score of experimental group was significantly higher than the groups of non-intervention,the difference was statistically significant (all P ＜ 0.05).While FL score of control group was not statistically significant before and after intervention (all P ＞ 0.05);Compared between two groups,there were statistical differences at 7 days,14 days in FL score (all P ＜ 0.05).In the experimental group,the histopathological changes of murine corneal epithelial cells had disorderly arrangement,increased layers of cells,and the inflammatory cells.Under electron microscope,the morphology of corneal epithelial surface cells was irregular and partially detached.The number of microvilli,desmosomes and hemidesmosomes were significantly decreased when compared to the control group.The expression of p-EGFR in experimental group was significantly less than that in control group,the difference was statistically significant (P ＜ 0.05).Conclusion Erlotinib can damage the tissue structure of corneal epithelium and ultrastructure of corneal epithelial cells in mice.And the mechanism is probably that erlotinib influence the corneal epithelium by inhibiting the EGFR activation.

Recent advances in the diagnosis and treatment of pediatric genetic cholestasis / 中华实用儿科临床杂志

Cholestasis is defined as a conjugated bilirubin level >1 mg/dL(17.1 μmol/L)if total serum bilirubin is ≤5 mg/dL(85.5 μmol/L),or conjugated bilirubin fraction >20％of total bilirubin when the total bilirubin is >5 mg/dL(85.5 μmol/L).In the recent years,the diagnosis and management of genetic cholestasis have caused considerable attention in the pediatric world,in pace with the development,maturation,and clinical application of the theories and techniques in genomics as well as molecular genetics.With a diversity of causative genes,genetic cholestasis usually demonstrates nonpathognomonic clinical manifestations.The etiology diagnosis such a disease relies on genetic tests,the treatment is often difficult,and the prognosis varies disparately,usually causing tremendous pain and burden on the affected patient and the family as well.Taking citrin deficiency,mitochondrial DNA depletion syndrome,microvi-llus inclusion disease and sodium taurocholate cotransporting polypeptide deficiency as samples,the recent advances in the diagnosis and treatment of genetic cholestasis are addressed.

Efeito da adição de agentes tróficos na dieta de leitões desmamados sobre a estrutura e ultraestrutura do intestino delgado e sobre o desempenho / The effect of the addition of trofic agents in weaned piglet diets over the structure and ultra-structure of small intestine and over performance

Foram estudados os efeitos da glutamina, dos ácidos graxos poli-insaturados e da parede celular de levedura (PCL) sobre a estrutura e ultraestrutura do intestino delgado e o desempenho de leitões. Foram utilizados 45 leitões, desmamados aos 21 dias de idade, para testar os seguintes tratamentos: T1 - dieta basal; T2 - dieta basal + 1 por cento de glutamina; T3 - dieta basal + 0,2 por cento de PCL; T4 - dieta basal + 5 por cento de óleo de peixe. Nos dias sete e 14 pós-desmame, foram abatidos cinco leitões de cada tratamento. Os aditivos testados não alteraram a altura e a densidade dos vilos nem a profundidade das criptas do intestino delgado. Foi observado efeito de idade, mostrando redução na altura e na densidade dos vilos e na profundidade das criptas após o desmame. No duodeno e jejuno, foram observados maiores valores de relação vilo:cripta, que aumentaram com a idade pós-desmame. Ocorreram redução da altura dos microvilos do duodeno aos sete dias e aumento da largura dos microvilos do jejuno aos 14 dias pós-desmame. A área de superfície apical dos enterócitos não foi alterada pelos fatores estudados. Os aditivos estudados não foram eficientes em prevenir a atrofia da mucosa intestinal do jejuno, ao não interferir na sua ultraestrutura. Os aditivos incluídos na dieta não influenciaram o desempenho dos leitões no pós-desmame.

The effects of glutamine, poliunsatured fatty acids and cellular wall of yeast (CWY) under the structure and ultra structure of the small gut and the performance of the piglets were studied. Forty five piglets weaned at 21 days were used to test the following treatments: T1 - basal diet; T2 - basal diet + 1 percent of glutamine; T3 - basal diet + 0,2 percent of CWY; T4 - basal diet + 5 percent of fish oil. At seven and 14 post weaning days, five piglets of each treatment were slaughtered. The height, density of villus and depth of small gut crypts were not altered by the inclusion of additives. The effect of age was observed, showing a reduction in the height and density of villus and depth of crypts after weaning. In duodenum and jejunum higher values were observed in the relation villus:crypt, which increased with the post wean age. There was a decrease in the height of microvillus of the duodenum at 7 days and an increase of the width of the microvillus of jejunum at 14 days after wean. The area of the apical surface of the enterocytes was not altered by the studied factors. The studied additives were not efficient to prevent the atrophy of the intestinal mucosa of the jejunum, since they did not interfere on its ultra structure. Piglet performance was not affected by the additives included in the diet.

Measuring the elasticity of HL-60 cell microvillus by parallel plate flow chamber / 医用生物力学

Objective To filter the noises in the experimental data of parallel plate flow chamber for observing more clearly the events occurring in the process of cell rolling adhesion and develop a new method to measure the elasticity of microvillus on cells based on the flow chamber experiment. Method The experiment of E-selectin regulated HL-60 cell rolling was performed by flow chamber system, and the data were denoised by wavelet analysis so that the high frequency thermal response signals were extracted from the data. Based on the equipartition theorem and equilibrium equations of tethered cell, the relationship between the cell microvillus spring constant and thermal fluctuations was constructed. Results Filtering noises from cell rolling time course by wavelet analysis, the events such as free rolling, slowing down, stopping and speeding up of rolling cell could be observed more easily; almost 80% of fluctuating energy of a rolling cell was involved in its high frequency fluctuation which was regarded as the thermal response of the cell to the Brown movement of water molecules, and the spring constant of microvillus on HL-60 cell was measured to be (13.7±7.4) μN/m at wall shear stress from 0.01～0.06 Pa. Conclusions The wavelet analysis can filter the thermal noises in cell rolling data of flow chamber experiment, and since the rigidity information of cell microvillus is involved in and can be extracted from the high frequency thermal fluctuation of the rolling cell, the parallel plate flow chamber experimental technique can be extended to measure the elasticity of microvillus on cells.