Establishing an experimental model of photodynamic induced anterior ischemic optic neuropathy

BACKGROUND： Scholars have supposed to establish animal models of optic neuropathy by pressing and partially amputating optic nerve, increasing intraocular pressure and injecting vasoconstrictor, etc., but the models are greatly different from anterior ischemia optic neuropathy. Therefore, a more ideal method is needed to establish animal model of anterior ischemic optic neuropathy （AION）. OBJECTIVE ： To establish AION models in rats, observe the functional changes of fundus, fundus fluorescein angiography （FFA）, optical coherence tomography （OCT）, flash visual evoked potential （F-VEP）, and histopathologically confirm its reliability. DESIGN： A randomized control tria SETTINGS ： Department of Ophthalmology, Xi＇an Fourth Hospital; Xi＇an Institute of Ocular Fundus Diseases MATERIALS ： The experiments were carried out in the research room of Xi＇an Institute of Ocular Fundus Diseases from February 2005 to May 2006. Thirty healthy male SD rats of 4-5 weeks old, weighing 140-160 g, were provided by the animal experimental center of the Fourth Military Medical University of Chinese PLA [SCXK （Military）2002-005], and those without eye disease examined by slit lamp and direct ophthalmoscope after mydriasis were enrolled. The conditions for feeding mice without special pathogen were strictly followed. The rats were randomly divided into blank control group （n =5）, laser group （n =5）, hematoporphyrin derivative （HPD） group and AION group （n =15）, each group was numbered randomly. For each rat, the right eye was taken as the experimental eye, and the left one as the control one. METHODS： In the AION group, the rats were injected with HPD （10 mg/kg） via caudal vein, and then the optic discs were exposed to krypton red （647 nm, 80 mV） for 120 s, and the rats were in avoidance of light for 2 weeks postoperatively. Rats in the laser group were only exposed to krypton red （647 rim, 80 mV） for 120 s, and in avoidance of light for 2 weeks postoperatively; Those in the HPD group were only injected with HPD （10 mg/kg） via caudal vein; Those in the blank control group were untouched. （1） Visual electrophysiological test： The F-VEP was used to evaluate the function of visual nerve. （2） FFA： After mydriasis and anesthesia as describe above, the fluorescein sodium parenteral solution （1 mL/kg） was injected v/a caudal vein and finished within about 3 s, the time of FFA was recorded from the beginning of injection, the video sight aimed at the optic disc and the surrounding area. （3） After mydriasis and anesthesia as describe above, the rats were examined with OCT. （4） Histological observation： After hematoxylin and eosin （HE） staining, the optic disc and surrounding blood vessels of retina were observed under light microscope at high power field. MAIN OUTCOME MEASURES： The results of fundus, FFA, visual electrophysiological test and OCT detection within 90 days after model establishment were observed. RESULTS： Of the 30 rats, 1 died after anesthesia in the laser group and 2 died in the AION group respectively, and finally 27 rats were involved in the analysis of results. （1） Changes in fundus： In the AION group, there was edema in upper optic disc and unclear boundary at 1 day after establishment, edema still could be observed at 6 days, and upper optic disc atrophied and appeared as pale at 90 days. （2） FFA results： In the AION group, early ＂low fluorescence＂, middle and late ＂high fluorescence＂ were observed in upper optic disc 1 day after model establishment, and there was ＂low fluorescence＂ at 6 days, and the low fluorescence could be observed all the time at 23 days. （3） Visual electrophysiological changes： In the AION group as compared with the control eyes, the experimental ones had prolonged F-VEP P100 latency [（71.65±8.81）, （57.58±8.38） ms, t =3.148, P =0.012], and decreased wave amplitude [（4.77±1.90）, （10.06±3.66） μV, t =4.082, P =0.003], and these changes lasted to 35 days after model establishment. （4） OCT results： In the AION group, the reflection surface of part nerve fiber layer was higher than the retina plane, the surface was rough and the thickness was increased at 6 days after model establishment. （5） Histopathological results： At 1 day after model establishment, part optic discs had highly edema, edema of nerve fibers, and loose tissue, also accompanied by the displacement of surrounding retina; At 23 days, the optic disc and surrounding nerve fiber layers became thinner, and the numbers of ganglion nuclei in the retina tissue sections were obviously decreased. These changes were not observed in the laser group, HPD group and blank control group. CONCLUSION ： The changes of fundus, FFA, OCT, visual electrophysiology and histopathology confirmed that the krypton red laser irradiation （647 nm） at 2 hours after HPD was injected via caudal vein can establish more ideal animal models of AION.

CT examination, clinical situation and experimental characteristics of infants with intracranial hemorrhage induced by delayed vitamin K deficiency

BACKGROUND: Delayed vitamin K deficiency is characterized by acute onset, severe illness and high fatality rate. 33%-50% survivors accompany with other various nervous system sequelas. Therefore, diagnosis and treatment of intracranial hemorrhage in time become a key factor for improving healing rate and reducing fatality rate and incidence of sequela. OBJECTIVE: To investigate the clinical situation, experimental characteristics, CT examination and terminative characteristics of infants with intracranial hemorrhage induced by delayed vitamin K deficiency. DESIGN: Case analysis. SETTING: Department of Pediatrics, Zhongshan Hospital Affiliated to Xiamen University. PARTICIPANTS: A total of 17 infants with intracranial hemorrhage induced by delayed vitamin K deficiency aged 1-3 months including 11 boys and 6 girls were selected from Department of Pediatrics, Zhongshan Hospital Affiliated to Xiamen University from January 1994 to December 2005. All infants had drowsiness, rejective milk, spiting milk, gaze of both eyes, tic, coma, full anterior fontanelle, high muscular tension and cerebral hernia, etc. Experimental examination demonstrated that infants had anemia at various degrees; prothrombin time and partial thromboplastin time were prolonged; platelet count was normal. CT examination indicated that screenages of subarachnoid hemorrhage, subdural hematoma, cerebral parenchyma hemorrhage and intraventricular hemorrage were changed. Hemorrhage was stopped by the application of vitamin K. All patients provided informed consent. METHODS: ① Clinical situation and physical sign of infants were observed after hospitalization and scanned with rapid spiral CT scanning system. The thickness and average space of layers were 8-10 mm and the scanning time was 5 s with window width of 30-80 Hu and window position of 28-35 Hu. ② After hospitalization, four items of blood coagulation was measured with Futura meter and biochemical indexes of blood, such as serum calcium, serum alkaline phosphatase, total bilirubin, direct bilirubin and indirect bilirubin, were detected with Roche Modular PPI automatic biochemistry analyzer. ③ After hospitalization, infants were given 5-10 mg vitamin K1 for 3-5 days, and then, they were transfused with 10-15 mL/kg fresh plasma or whole blood for 1-3 times and received other relative therapies. Therapeutic effects were evaluated based on clinical diagnosis and criteria. MAIN OUTCOME MEASURES: ① Clinical situation, physical sign and CT examination; ② results of experimental examination; ③ treatment and termination. RESULTS: A total of 17 infants with intracranial hemorrhage were involved in the final analysis. ① Clinical situation, physical sign and CT examination: All infants had pale facial expression and full or bossing anterior fontanelle. Among them, 13 infants had drowsiness or dysphoria, 12 rejective milk or emesis, 11 tic, 13 injection site hemorrhage, 2 gastrointestinal hemorrhage, 3 cerebral hernia, 11 high muscular tension and 6 cervical rigidity. CT examination demonstrated that most infants (88%, 15/17) had subarachnoid hemorrhage; 10 (59%, 10/17) had subdural hematoma; 8 (47%, 8/17) had cerebral parenchyma hemorrhage; few had intraventricular hemorrhage. In addition, results of CT examination also indicated that 17 infants had intracranial hemorrhage. Hemorrhage sites of 12 infants were equal to or more than 2, which was accounted for 70% (12/17); meanwhile, partial cases accompanied with a large area of focus of cerebral infarction or cerebral hypoxia-ischemia lesion. ② Results of experimental examination: There were 6 infants with elongation of prothrombin time, 5 with partial elongation of prothrombin time, 4 with decrease of serum calcium (1.69-2.25 mmol/L), 3 with increase of serum alkaline phosphatase, 3 with increase of total bilirubin, 3 with increase of direct bilirubin, and 3 with increase of indirect bilirubin. ③ Treatment and termination: After treatment, 12 infants were cured well, 3 improved, 1 given up and 1 died. Later, ten infants received CT re-examination at 3 months after treatment. The results indicated that 3 infants had simple subarachnoid hemorrhage and 4 had subarachnoid hemorrhage accompanying with subdural hematoma. Their focuses were absorbed well and not show as obvious sequela. One infant had subdural hematoma accompanying with subarachnoid hemorrhage, cerebral parenchyma hemorrhage, intraventricular hemorrage and cerebral hypoxia ischemia, and then, after hematom absorption, obvious cerebral malacia focus, hydrocephalus, brain atrophy and inferior accumulating fluid of dura mater were observed; 2 had subdural hematoma accompanying with subarachnoid hemorrhage and cerebral hypoxia ischemia, and then, after bleeding absorption, brain atrophy was changed remarkably; changes of hydrocephalus were observed in one infant. CONCLUSION: ① Symptoms of pale facial expression, full or bossing anterior fontanelle, drowsiness and dysphoria are observed in infants with delayed vitamin K deficiency . ② Experimental indexes demonstrate that prothrombin time and partial prothrombin time are prolonged, and numbers of infants having decrease of serum calcium are in the third place. ③ Poly-intracranial hemorrhage is a notable characteristic of CT examination. Partial infants who have poly-intracranial hemorrhage always accompany with cerebral hypoxia-ischemia lesion or cerebral infarction. Clinical situation and prognosis of infants who have a large area of intracranial hemorrhage and cerebral hypoxia-ischemia changes are poor; however, those of infants who have simple subarachnoid hemorrhage or combination of subarachnoid hemorrhage with subdural hematoma are well. ④ Effect of vitamin K on this kind of disease is well.

Experimental repetitive mild traumatic brain injury induces deficits in trabecular bone microarchitecture and strength in mice

To evaluate the long-term consequence of repetitive mild traumatic brain injury （mTBI） on bone, mTBI was induced in 10-week-old female C57BL/6J mice using a weight drop model, once per day for 4 consecutive days at different drop heights （0.5, 1 and 1.5 m） and the skeletal phenotype was evaluated at different time points after the impact. In vivo micro-CT （μ-CT） analysis of the tibial metaphysis at 2, 8 and 12 weeks after the impact revealed a 5%-32% reduction in trabecular bone mass. Histomorphometric analyses showed a reduced bone formation rate in the secondary spongiosa ofl.5 m impacted mice at 12 weeks post impact. Apparent modulus （bone strength）, was reduced by 30% （P 〈 0.05） at the proximal tibial metaphysis in the 1.5 m drop height group at 2 and 8 weeks post impact. Ex vivo μ-CT analysis of the fifth lumbar vertebra revealed a significant reduction in trabecular bone mass at 12 weeks of age in all three drop height groups. Serum levels of osteocalcin were decreased by 22%, 15%, and 19% in the 0.5, 1.0 and 1.5 m drop height groups, respectively, at 2 weeks post impact. Serum IGF-I levels were reduced by 18%-32% in mTBI mice compared to control mice at 2 weeks post impact. Serum osteocalcin and IGF-I levels correlated with trabecular BV/TV （r2 = 0.14 and 0.16, P 〈 0.05）. In conclusion, repetitive mTBI exerts significant negative effects on the trabecular bone microarchitecture and bone mechanical properties by influencing osteoblast function via reduced endocrine IGF-I actions.

Effect of metal surface morphology on nano-structured patterns induced by a femtosecond laser pulse and its experimental verification

The effect of material surface morphology on the periodic subwavelength of nano-structures induced by a femtosecond（fs） laser was investigated systematically from the initial surface roughness, the different scratches, the pre-formed ripples, and the ＂layer-carving＂ technology experiments. The results of the comparative experiments indicate that the initial surface conditions of the target surface have no obvious effects on the spatial structured periods（SSPs） and the ripple orientation of the periodic nano-structures induced by a fs laser, which agreed well with the foretold present surface two-plasmon resonance（STPR） model. Furthermore, different shapes of nanogrids with high regularity and uniformity were obtained by fs-laser fabrication.