Background The establishment of diabetic animal model is a crucial step for the study about diabetic eye diseases. At present,the main modeling method include the injection of streptozotocin and alloxan. But the shortcoming of the former is an expensive price, and that of the later is high death rate of animals. Objective This experiment was to discuss the way which decrease the death of alloxan-injected animal and explore the effects of high blood glucose on the posterior capsular opacification (PCO). Methods Forty clean healthy male New Zealand white rabbits were randomly divided into 2 groups. 90mg/kg of alloxan were injected via ear vein once in 20 rabbits to create the diabetic animal models,and the equivalent amount of normal saline solution was injected at the same way as normal blood glucose group. The successful models were selected in the animals with the blood glucose level over 12. 0 mmol/L two weeks later, and PCO of lens were graded based on the method of Odrieh under the slit lamp. Extracapsular lens extraction was then performed on the right eye of rabbits in both groups, and the posterior capsules were obtained from these eyes at the 6th, 10th and 14th days after operation. The expression of proliferating cell nuclear antigen ( PCNA ) in posterior capsular lens epithelial cell was detected by immunohistochemistry. Results The modeling successful rate was 70% after injection of alloxan. The body weight of rabbits in high blood glucose group was significantly lowed and the blood glucose was significantly elevated in comparison with normal blood glucose group ( all P<0. 05). Two weeks after surgery ,2 eyes occurred 2 grade of PCO and only one eye showed the 1 grade of PCO in the high blood glucose group. However, 1 grade of PCO was found in 3 eyes in the normal blood glucose group. Biopsy revealed that PCNA was positively expressed in the cell nuclei of LECs in high blood glucose group rather than the normal blood glucose group from the 10th day after surgery. The proliferation index of PCNA was 0. 86±0. 04 and 0. 25±0. 03 respectively in high blood glucose group and normal blood glucose group, showing a significant difference between them (t = -16. 171 ,P = 0. 000). Conclusion Stable diabetic models of rabbits can be created by intravenous injection of 90 mg/kg alloxan. High blood glucose level is one of the important factors for the development of PCO.

Objective To observe the effects of thin recipe ofBuyang Huanwu Decoction on expressions of glutathione (GSH) antioxidant system including GSH, GSH-Px andγ-GCS in the brain of focal cerebral ischemia rats;To study its mechanism of antioxidant effect.Methods Totally 120 SD rats were randomly divided into four groups according to random number table method:sham-operation group, model group,Buyang Huanwu Decoction group and thin recipe ofBuyang Huanwu Decoction group. Except for the sham-operation group, the rest groups established focal cerebral ischemia rat model by middle cerebral arterial occlusion. The treatment groups were given corresponding medicine by gavage, while the sham-operation group and model group were given the same amount of normal saline 2 h after modeling. Each group was detected after cerebral ischemia for 1 day, 3 days, and 7 days respectively. The content of GSH and the activity of GSH-Px were detected. At the same time,γ-GCS mRNA and protein levels were determined by using real time-PCR and Western blot.Results The content of GSH and the activity of GSH-Px at each time point decreased in the model group (P<0.05), compared with the corresponding sham-operation group. But the expression ofγ-GCS mRNA and protein increased, with statistical significance on day 1 (P<0.05). Compared with model group, content of GSH and GSH-Px activity levels increased differently in each treatment group, whileγ-GCS mRNA and protein expression raised, with statistical significance on day 3 (P<0.05).Conclusion The thin recipe ofBuyang Huanwu Decoction plays antioxidant effect by regulating the expression of glutathione antioxidant system GSH, GSH-Px andγ-GCS after cerebral ischemia, which may be one of its therapeutic mechanisms for cerebral ischemia.

Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO.

Objective To observe the effects of simplified recipe ofBuyang Huanwu Decoction on expression of APJ in the brain of local cerebral ischemia rats;To discuss its mechanism of action. MethodsFocal cerebral ischemia rat models were established by middle cerebral arterial occlusion. The adult rats were randomly divided into sham-operation group, model group,Buyang Huanwu Decoction group and simplified recipe ofBuyang Huanwu Decoction group. Administration groups were given relevant medicine for gavage. The expressions of APJ protein and APJ mRNA at different time points were detected by Western blot and RT-PCR.ResultsCompared with model group and sham-operation group, the expression of APJ protein and APJ mRNA at different time points in Buyang Huanwu Decoction group and simplified recipe ofBuyang Huanwu Decoction group significantly increased (P<0.05). The expression of APJ protein at different time points showed no difference between simplified recipe ofBuyang Huanwu Decoction group andBuyang Huanwu Decoction group;while the expression of APJ mRNA in simplified recipe ofBuyang Huanwu Decoction group was higher than Buyang Huanwu Decoction group (P<0.05).ConclusionSimplified recipe ofBuyang Huanwu Decoction plays a role in neural protection and restoration by promoting the expression of APJ in the brain of focal cerebral ischemia rats.

Background Efficient and lowcost way to isolate keratocytes is helpful for research on cornea.Either relatively expensive or inefficient is the shortage of those means now applied,while raising the keratocytes through passage will change the phenotype of them quickly.Our aim is to approach the way getting keratocytes effectively utilizing modified two step enzymatic digestion by type I collagenase. Objective To evaluate the effect of isolating the bovine keratocytes utilizing two step enzymatic digestion and observe the morphological changes of the keratocytes during cultivation in vitro. Methods Keratocytes were isolated from bovine corneas using 0.5 mg/mL and 1 mg/mL type I collagenase digestion.The harvesting rate and viability rate of the primary keratocytes were evaluated.During the primary cultivation in vitro,the morphological changes of the keratocytes and their F-action distribution were observed.Results(1)The extracellular matrix of the bovine corneas were almost dissolved by the two step enzymatic digestion,followed the keratocytes completely isolated from the solid matrix.The amount of the harvested keratocytes from each cornea was(2.11±0.15)X106 on average while the viability rate was(91.69±3.73)% and the inoculation rate Was(81.20±1.25)%.(2)The primary keratocytes attached and spreaded out with dendritic and stellate morphology.After 3 days cultured,the branches of the keratocytes were contacting and formed networks.The F-actin detected by phalloidin binding showed a limited cortical localization. Conclusion (1)The method of two step enzymatic digestion can make the extracellular matrix of bovine cornea stroma completely degraded with the advantages in high efficiency of harvesting keratocytes and high cell viability and relatively simple manipulation. (2) The primary bovine keratocytes have dendritic morphology and with limited F-action distribute in cellular cortex.

OBJECTIVETo investigate the technique and clinical effect of interlocked intramedullary nailing through a single incision on the treatment of floating knee injuries.

METHODSThirty-eight patients of floating knee injuries included 25 male and 13 female with an average age of 35.6 years ranging from 17 to 52 years. All patients were treated by interlocked intramedullary nailing through a single incision. The clinical effect was evaluated postoperatively.

RESULTSThirty-eight cases were followed-up for 8.6 to 15.4 months with an average of 12.0 months. Thirty-seven cases gained bone healing and the time of fracture healing was from 2.7 to 6.9 months, with an average of 4.8 months. The nonunion of the distal femoral fracture was observed in 1 case with retrograde nailing, the revision operation was performed by plate and bone graft, and bony union was obtained. The results were excellent in 22 cases, good in 13 cases, fair in 2 cases, and poor in 1 case.

CONCLUSIONThe treatment of floating knee injuries with interlocked intramedullary nailing through a single incision brings less damage, low infection rate, good fracture healing. It is an effective method for treating floating knee injuries.

Adolescent ; Adult ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; methods ; Humans ; Knee Injuries ; surgery ; Male ; Middle Aged

OBJECTIVETo study the result of less invasive fixation in treating comminuted fracture of distal tibia.

METHODSFrom 2002 to 2008, 48 patients with comminuted fracture of distal tibia were treated with surgery. The closed reduction and less invasive fixation were done to stable broken fibula, and the reduction by Kirschner wire to pick was used for relatively larger debris of tibia. The Kirschner wire or screw were used to fix fracture after the restore of the ankle cavity position and the alignment of the tibia. Partial weight loading and functional exercise of ankle joint were done at 6th week after operation.

RESULTSForty-eight patients were followed up for 1-24 months with an average of 12 months. All the fractures were united. According to Johner-Wruhs standard to value the result by factors of pain, deformity, motion range of joint,with or without injury of nerve and blood vessel. Thirty-eight cases obtained excellent result, 8 good, fair 2. The rate of excellent and good were 95.8%.

CONCLUSIONLess invasive fixation has ascendancy such as easy operation, less injury of soft tissue, reliable fixation, which can maximally protect periosteum. It is a choice for treating comminuted fracture of distal tibia.

Adult ; Aged ; Bone Wires ; Female ; Fracture Fixation, Internal ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Tibia ; injuries ; surgery ; Tibial Fractures ; surgery ; Young Adult

OBJECTIVETo compare the clinical efficacy of 3 approaches of vertebroplasty in the treatment of severe osteoporotic vertebral compression fractures.

METHODSTwenty-five patients with severe osteoporotic vertebral compression fractures were observed, whose average age was 72 years with average disease history of 12 days and average compression of the affected vertebral bodies of 73%. The patients were divided into 3 groups for 3 different fracture types according to Rao's classification of osteoporotic vertebral structure and deformity, namely wedge type (group A, n= 12), biconcave type (group B, n= 7) and crush type (group C, n=6). Unipedicular approach was adopted in group A, far lateral bipedicular approach in group B, and posterior wall vertebroplasty and pedicle screw fixation in group C. The average follow-up time was 1 year. Visual analog scale (VAS), analgesic use and the mobility were measured in the patients preoperatively and 3 days (7 days in group C) and 6 months after the operation, respectively, and the success rates and complications were observed.

RESULTSAll the surgical procedures were successful. The average operation time was 35 min in group A, 50 min in group B, and 2 h in group C. The average volume of cement injected into each vertebral body was 2.0 ml. The average blood loss was 30 ml in groups A and B, and 600 ml in group C. The procedure increased mobility and decreased analgesic use. VAS was decreased by a mean of 4.8 in Group A, 6.2 in group B 3 days after the operation and 5.4 in group C 7 days postoperatively, and remained stable till 6 months after the operation (P<0.01). Cement extravasation occurred in 9 cases, cement in the vertebral canal in 2 cases with transient neurological symptoms, and cement extravasation in the intervertebral space and the anterior space of the vertebral bodies took place in 5 and 2 cases, respectively. No neurological complications were observed.

CONCLUSIONDifferent types of severe osteoporotic vertebral compression fractures require management with different approaches of vertebroplasty for adequate filling of the remaining vertebral body, which provides significant pain relief with wider indications.

Aged ; Female ; Fractures, Compression ; etiology ; surgery ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Osteoporosis ; complications ; Spinal Fractures ; etiology ; surgery ; Thoracic Vertebrae ; injuries ; Treatment Outcome ; Vertebroplasty ; methods

Objective To observe the brain activation of interictal epiletiform discharges(IEDs) and to localize the epileptogenic foci of epilepsy.Methods The electroencephalogram(EEG)and functional MRI data of 12 focal epileptic patients were acquired using a combination of EEG and functional MRI simultaneously.The IEDs onset time detected with EEG were set as the time parameters in an event- related paradigm of functional MRI analysis.The spatial and temporal characters of IEDs activation were analyzed in detail.In order to confirm the consistency of this method,all patients were scanned repeatedly and the results were correlated with clinical evaluation.Results Of the 12 patients,valid data from EEG- fMRI were obtained from 10 patients in a total of 18 sessions..Compared with the structural foci,the epileptic foci localization results of eleven sessions were good,five sessions were fairly good,and two sessions were poor.The results obtained from six patients in two separate sessions were concordant.respectively.Moreover,thalamic activation was detected in ten sessions,cerebellar activation was detected in all sessions,and the deactivation was found in the default mode loci in nine sessions. Conclusion The method of performing EEG and fMRI simultaneously can potentially be a useful tool in epilepsy research.