Objective To explore the mechanism of pulmonary hypoplasia in case of congenital diaphragmatic hernia (CDH), and study the ultramicrostructural features of lung tissue of CDH fetal rat models at different developmental stages. Methods Seven SpragueDawley (SD) pregnant rats were randomly divided into CDH group (n=4) and control group (n=3). For the rats in the CDH group, Nitrofen was used to fill in the stomach once at day 9.5 of pregnancy (125 mg of Nitrofen dissolved in 2 ml of olive oil each), and 3, 10, 17 fetal rats were collected at day 16, 18 and 21 of pregnancy respectively. For the rats in the control group, 2 ml of olive oil was used to fill in the stomach, and 10 fetal rats were collected at day 16, 18, and 21 of pregnancy respectively. The lung tissue sections of the fetal rats collected on day 16 were observed under transmission electron microscope (TEM). For the lung tissue of the fetal rats collected on day 18, hematoxylineosin (HE) staining and TEM observation were performed and the incident of CDH was detected. Besides the procedures carried out for the rats collected on day 18, the ratio of fetal lung to body weight was observed for the lung tissue of the fetal rats collected on day 21. Results (1) The ratio of fetal lung to body weight of fetal rats in the CDH group was significantly lower than that of fetal rats in the control group (0.0238 vs. 0.0430, Plt;0.01). The incidences of CDH in the 18thday and 21stday fetal rats in the CDH group were 90.00% and 82.35%respectively, while no CDH was observed in the corresponding fetal rats in the control group, suggesting pulmonary hypoplasia in the CDH group. (2) The ultramicrostructural observation showed that compared with the control group, pulmonary hypoplasia appeared in 16thday fetal lungs in the CDH group, i.e., broad breathing barrier substrate, little contents, predominant euchromatin and rich ribosomes in the alveolar epithelial cells, and no microvilli in the bronchial lumen. The observation on the 18thday and 21stday samples suggested that, with the progressing of pregnancy, the abovementioned features became more obvious. (3) Typical lamellated body was observed in fetal lung type Ⅱ alveolar epithelial cells from the 21stday fetuses in both the CDH group and the control group, suggesting that some late subcellular structures were normal. Conclusion Lung hypoplasia develops in the early period of fetal rats with CDH rather than in the late period, implying that the treatment of pulmonary hypoplasia of diaphragmatic hernia should be performed in the early stage of lung development.
Objective To study the influence of ischemia-reperfusion on the expression of the hyperpolarization activated cycl icnucleotide gated cation channel 4 (HCN4) and to discuss the mechanism of functional disturbance of sinoatrial node tissue (SANT) after ischemia reperfusion injury (IRI). Methods Eighty five healthy adult rabbits, weighing 2-3 kg, were randomly divided into 3 groups: control group [a suture passed under the root section of right coronary artery (RCA) without l igation, n=5], experimental group A (occluding the root section of RCA for 30 minutes, then loosening the root 2,4, 8 and 16 hours, n=10), experimental group B (occluding the root section of RCA for 1 hour, then loosening the root 2, 4,8 and 16 hours, n=10). At the end of the reperfusion, the SANT was cut off to do histopathological, transmission electronmicroscopical and immunohistochemical examinations and semi-quantitative analysis. Results The result of HE stainingshowed that patho-injure of sinoatrial node cell (SANC) happened in experimental groups A and B after 2 hours of reperfusion, the longer the reperfusion time was, the more serious patho-injure of SANC was after 4 and 8 hours of reperfusion, SANC reached peak of damage after 8 to 16 hours of reperfusion; patho-injure of SANC was more serious in experimental group B than in experimental group A at the same reperfusion time. Immunohistochemical staining showed that the expression of HCN4 located in cellular membrane and cytoplasm in the central area of SANC and gradually decreased from the center to borderl ine. The integral absorbance values of HCN4 expression in the control group (397.40 ± 34.11) was significantly higher than those in the experimental group A (306.20 ± 35.77, 216.60 ± 18.59, 155.40 ± 19.11 and 135.00 ± 12.30) and in the experimental group B (253.70 ± 35.66, 138.70 ± 13.28, 79.10 ± 9.60 and 69.20 ± 8.42) after 2, 4, 8 and 16 hours of reperfusion (P lt; 0.05). With reperfusion time, the expression of HCN4 of SANC decreased, which was lowest after 8 hours of reperfusion; showing significant difference among 2, 4 and 8 hours after reperfusion (P lt; 0.05) and no significant difference between 8 and 16 hours after reperfusion (P gt; 0.05). At the same reperfusion time, the expression of HCN4 was higher in the experimental group A than in the experimental group B. The result of transmission electron microscope showed that ultramicrostructure of SANC was damaged after reperfusion in experimental groups A and B. The longer the reperfusion time was, the more serious ultramicrostructure damage of SANC was, and reached the peak of damage after 8 hours of reperfusion. Ultramicrostructure of SANC was not different between 8 and 16 hours of reperfusion. At the same reperfusion time, the ultramicrostructure damage of SANC was moreserious in experimental group B than in experimental group A. Conclusion IRI is harmful to the morphous and structure ofSANC, and effects the expression of HCN4 of SANC, which is concerned with functional disturbance and arrhythmia.
rough the ultramicroscopic observation on muscle and microcirculation, Group A,where a largeamount of DXM combined with heporin was given svstematically and locally into the femoral artery of the severed limb before replantation, and in Group B only heporin was given, and Group C and D ascontrol.The results showed that if the hormone and heparin were administred in large dosage, it wasadvantageous to reduce the tissues from reperfusion injury during delayed replantation.