Objective Previous study showed that the histopathological basis of visual function damage caused by optical nerve injury is apoptosis of retinal ganglion cells(RGCs). This procedure is regulated by P53, bax and caspase 3 genes. Present study aimed to observe the expression of bax, P53 and caspase 3 mRNA in RGCs after traumatic optic nerve damage in the rats by SYBR green I fluorescence quantitative PCR method. Methods The animal model of optic nerve injury was established in the right eyes of 56 adult Wistar rats by a fluid percussion brain injury device (FPI) . Animal were killed on days 1, 3, 5, 7, 9, 14, 28 days separately after injury. Other 16 Wistar rats were divided into normal control group and sham operation group. The total RNA was isolated from rat fresh retina tissue by Trizol method and was treated by reverse transcription to cDNA using 01igo(dt) 18 as primer and then amplified. The target fragments of bax, P53 and caspase 3 cDNA were linked with carrier pTZ57 R/T to construct recombined plasmids which were transformated to E. Coli DH5α by T/A clone method. Recombined plasmids were extracted with alkaline lysis method and the plasmids were selected in white colonies by ampicillin screening, EcoR I restrictive enzyme analysis, and their specificity was evaluated using DNA sequencing. The standard curves were created by plasmid DNA and the precise expression level of target genes in samples were determined using software. The results were expressed as the ratios of target gene mRNA to GAPDH mRNA. Results The standard curve drawn by pTZ57R/T and target gene presented a good linear tendency with the higher sensitivity and specificity. The expression of P53 and bax mRNA began to increase on the third day after the injury of optic nerve and peaked on the fifth day and started to decline on the seventh day. The expression of caspase 3 mRNA increased from the fifth day through the ninth days after injury and declined on the fourteenth day. The significant differences were found in the expression of P53, bax and caspase 3 between model group and control group (P ＜ 0. 05) . Conclusion The pro-apoptotic protein P53, bax and caspase 3 play an important role in RGCs apoptosis.

BACKGROUND:The clinical research have found that the interbervebral disc herniation often occurs in several members or even al the members of a family, and the location, reason and symptom are basical y the same, indicating that genes play an important role in this kind of disease. OBJECTIVE:To analyze the apoptosis Fas gene expression characteristics of lumbar disc in the familial patients with intervertebral disc herniation. METHODS:Semi-quantitative reverse transcription-PCR was used to test Fas gene expression of vertebral pulp and cartilage endplate in the intervertebral disc among 15 familial patients, 21 ordinary patients and five fresh cadavers. RESULTS AND CONCLUSION:Fas gene expression level of endplate of familial and ordinary patients with intervertebral disc herniation was higher than that of fresh cadavers, and there was no significant difference (P<0.05);there was no significant difference in Fas gene expression in endplates between familial patients and ordinary patients with intervertebral disc herniation (P>0.05). Compared with the vertebral pulps of ordinary patients with intervertebral disc herniation and fresh cadavers, there was no significant difference in the Fas expression of vertebral pulps of familial patients with intervertebral disc herniation (P>0.05). The increasing Fas gene expression may be secondary in the endplates of familial patients with intervertebral disc herniation, which can prevent intervertebral disc degeneration through preventing the endplate degeneration.

Background Visual electrophysiology is a sensitive index for the evaluation of visual function.It has an important value in the assessment of traumatic optic neuropathy.Rabbit is an ideal animal model of traumatic optic neuropathy,and it is simple for the record of flash visual evoked potential(F-VEP)in rabbits.ObjectiveThe present study is to establish the animal model of traumatic optic neuropathy with or without lens injury and observe the repairing procedure using F-VEP. MethodsModels of traumatic optic neuropathy associated with lens injury were established in the right eyes and only traumatic optic neuropathy were created in the left eyes of 64 healthy SPF Chinese white rabbits using fluid percussion brain injury device(FPI).F-VEP was recorded based on the Proposal of International Visual Electrophysiology on 1,2,4,7,10,14,21,28 days after injury of optic nerves.Experimental animals were sacrificed in above time points for the histopathological examination.Macrophages were labeled by ED-1 antibody and survival retinal ganglion cells (RGCs)were stained by Nissl method.Results At the first day after injury,the latencies of P_(100) in both group were longer,and the amplitudes of P_(100) in both group were lower than before injury,showing statistically significant differences among different time points(P<0.05),but no significant difference was seen between the two groups(P>0.05).The duration of latency in traumatic optic neuropathy associated with lens injury group was shorter than that in only traumatic optic neuropathy group(P<0.05).The restore of latency in traumatic optic neuropathy associated with lens injury group was much faster than that in only traumatic optic neuropathy group(P<0.05).The numbers of macrophages were significantly increased and numbers of survival RGCs were considerably decreased with lapse of injury time (P<0.05).The abnormalities of VEP P_(100) and RGCs were obviously improved in 28 days after injury in both groups. ConclusionThis animal model can be established successfully by FPI.The result of retinal histopathological examination confirms F-VEP findings in this model.

Hypoxic-ischemic brain damage (HIBD) is referred to a common type of cerebral damage, which is caused by injury, leading to shallow bleeding in the cortex with intact cerebral pia mater. In recent years, studies show that a various kinds of immune cells and immune cellular factors are involved in the occurrence of HIBD. CC chemokine receptor 2 (CCR2) is a representative of CC chemokine receptor, and is widely distributed in cerebral neuron, astrocyte, and microglial cells, and is the main chemo-tactic factor receptor in brain tissue. CC chemokine ligand 2 (CCL2) is a kind of basophilic protein and the ligand of CCR2, and plays an important role in inflammation. In order to provide evidence for correlational studies in HIBD, this review will introduce the biological characteristics of CCR2 and CCL2, and illustrate the relationship between the immunoreactivity and HIBD.
Animals ; Brain Injuries/pathology* ; Cerebral Cortex/physiopathology* ; Chemokine CCL2/metabolism* ; Chemokines, CC/metabolism* ; Hypoxia-Ischemia, Brain/metabolism* ; Macrophage Inflammatory Proteins/metabolism* ; RNA, Messenger/metabolism* ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR2/metabolism*

Objective To prospectively evaluate the feasibility and reliablity of low kilovoltage contrast-enhanced multi-slice CT(MSCT)to detect the myocardial viability in chronic myocardial infarcetion.with comparison to magnetic resonance(MR)myocardial perfusion and viability imaging.Methods Thirty-two patients with clinical diagnosed chronic myocardial infarction underwent the first pass and delayenhanced myocardial imaging with 64-slice MSCT and MR Left ventricle was divided into 16 segments.MSCT and MR images of all the patients were blindly analyzed.The size and extent of hypoenhanced regions in first pass phase and hyperenhanced regions in delayed phase were define.The Kappa test was used to assess the ability of identifying the viable myocardium between the two methods.Results In 32 patients with chronic myocardial infarction,MSCT showed hypoenhanced regions in 41 segments and normal in 471 segments during the first phase,while MRI revealed hypoenhanced regions in 47 segments and normal regions in 465 segments.The Kappa value was 0.650 and the concordance rate of the two techniques was 94.5%(484/512).MSCT showed 135 hyperenhanced regions which were non-transmural in 50 segments and transmural in 85 segments.And the other 377 normal segments showed no enhancement in the delayed phase.MRI revealed 120 hyperenhanced regions which were non-transmural in 56 segments and transmural in 64 segments.And the other 392 normal segments showed no enhancement in the delayed phase.The Kappa value of the two techniques was 0.609 and the concordance rate of the two techniques was 80.7% (413/512).Conclusion The study showed that low kilovolrage CE MSCT has a good concordance with MRI and has high feasibility and reliability in evaluating the myocardial viability in chronic myocardial infarction.The radiation dose is still the important aspect of MSCT application.

AIM: To observe the effects of porous polyethylene ( Medpor) as a spacer graft in the reconstruction of large areas eyelid defect after the operation of malignant tumors of lower eyelids.
METHODS: Nineteen cases ( 19 eyes ) of malignant tumors of lower eyelid underwent the eyelid reconstruction were selected. Medpor lower eyelid inserts implantation were used to replace tarsal joint sliding conjunctival flap and pedicle flap, and repaired full -thickness lower eyelid defects then underwent eyelid reconstruction.
RESULTS: Appearance of eyelids and functional improvements were satisfactory with no stimulation on the eyeball and no effect on the visual function. Implants is with no absorption, shift, exclusion or infection and no tumor recurrence in all cases during the follow up for 6-36mo.
CONCLUSION: Medpor lower eyelid inserts implantation can instead tarsal plate for the reconstruction of medium to large areas lower eyelid defect, which is easy performing with rare complications. It is an ideal alternatives of tarsal plate.

Objective To investigate the multi-drug resistance of Klebsiella strains and its mechanism.Methods Twenty strains of Klebsiella were isolated from the Affiliated Hospital of Medical College,Ningbo University from October 2009 to March 2011,in which 18 isolates were Klebsiella pneumonia and 2 were Klebsiella planticola. Drug sensitivity was determined by K-B tests. Drug resistant genes gyrA,parC (chromosome mediated) and aac( 6′)-I b-Cr,qnrA,qnrB,qnrS,qepA (plasmid mediated) were amplified by PCR and verified by direct automated fluorogenic sequencing. Results Resistance to β-1actams,aminoglycosides and quinolones was observed in 20 strains,and resistant rates were all above 80％.Klebsiella planticola strains were sensitive to imipenem and meropenem.Mutations of gyrA and parC genes existed in 18 strains (90％),and the positive rates of aac (6') -I b-C r,qnrB and qnrS were 60％ (12/20),20％ (4/20) and 20％ (4/20),respectively.Conclusion The mutations ofgyrA and parC genes may be the main cause of the resistance to quinolones in these strains.

Objective To establish a stable cell line secreting human IgE Cε2-4 protein, and in-vestigate the binding capacity of receptor FcεR Ⅰ Methods The E24 gene was derived from SKO-O07 cell line, and was then cloned into pcDNA3.1 (+) (signal peptides were synthesized and fused at the 5'-end of E24 gene) or pCMV-L vector. After transient transfection into 293T cell, the secreted F24 protein was ana-lyzed by sandwich ELISA. The best vector was chosen to be transfected into CHO cells with LipofectAMI-NETM 2000 reagent. After being selected by G418 and subcloned three times by limited-dilution method, two stable cell lines were established. E24 gene was amplified by RT-PCR, and the E24 protein in the superna-tant was identified by ELISA. Besides, the binding capacity of FceR ⅠⅡ was analyzed by flow cytometry method. Results Three mammalian expression vector SP-E24-F3. 1, SP lI-E24-P3.1 and E24-PL were constructed and transient transfected to 293T cells. The output of E24 protein in the supernatant were 19.1, 19.4 and 8.7 μg/ml, respectively. Then the vector SP IX-E24-P3.1 was transfected into CHO cells. Final-ly, two single clones secreting E24 protein were stably obtained. The output of E24 were all at least 25 μg/ml. RT-PCR could detect the E24 gene from one of the two clones. Furthermore, flow cytometry results showed that E24 could bind the receptor in a dose-dependent manner. Conclusion Two stable cell line se- creting E24 protein were obtained, while E24 could specifically bind FcεR Ⅰ.

Objective To built up the ERP model,measure mode and P300 potential reference standard in mice with vascular dementia(VD),and characterize the P300 potential in mice with VD.Methods Fortyeight mice were randomly divided into a normal group.sham operation group and a VD group.The mice in the Vd group were subject to repetitive ischemia and reperfusion by using the ligation of bilateral common carotid arteries so as to establish the VD model.The behavioral abnormalities were investigated by step-down test and water maze test.The N2 and P3 components of P300 potentials were also recorded.Results It was shown that the learning and memory abilities as reflected by the step down test and water maze test scores were decrease in mice in the VD group when compared with those in the normal group and sham operation group(P<0.05).The N2 and P3 latencies significantly prolonged(P<0.01)and P3 amplitudes decreased(P<0.05)in VD group as well.Conclusions In VD mice,there is a significant prolongation of the P300 potential latency and a significant decrease of learning and memory abilities.Recordings of P300 from unanesthetized mice could be an objective,non-invasive,quantitative and valuable electrophysiological method for studying the cognitive function of VD mice.

Objective To evaluate low-dose CT coronary angiography with prospective electrocardiogram (ECG)-triggering using dual-source CT scanner.Methods Sixty-eight patients who underwent coronary CT angiography using a dual-source CT scanner were divided into 2 groups: group A (38 cases) and group B (30 cases).Prospective ECG-triggering sequence scan mode was employed for group A.Inclusion criteria included: heart rate <70 bpm, sinus rhythm, and heart rate fluctuation less than 10 bpm.Data acquisition was set at 70% of the RR-interval.Retrospective ECG-gating helical scan was performed for group B.Inclusion criteria included heart rates < 70 bpm and sinus rhythm.The exclusion criteria included heart failure and serious arrhythmias.In both groups, patients with a BMI≥24 kg/m2 were examined with a tube voltage of 120 kV, whereas patients with a BMI <24 kg/m2 were examined with a tube voltage of 100 kV.All images were transferred to a workstation for further processing and analysis.The imaging quality was evaluated.The imaging quality of coronary artery segments were compared with rank sum test between the two groups, and the radiation dose were compared with t test.Results A total of 476 coronary artery segments were evaluated in group A and 372 segments were evaluated in group B.The mean score of imaging quality for coronary artery segments in group A was 3.48±0.59 and that in group B was 3.53±0.58.There was no statistical difference in imaging quality between the two groups (Z=-1.432, P=0.187).The effective dose was on average (2.51±0.54) mSv (range 1.3--3.3 mSv) in group A, whereas on average (14.55±3.54) rosy (range 7.1--20.2 mSv) in group B.There was a statistical difference between the two groups (t=18.484, P=0.000).Conclusions Low-dose prospective ECG-triggering sequence scan in dual-source CT coronary angiography is feasible in patients with low heart rate and regular cardiac rhythm.This scan mode can substantially reduce radiation doses while preserving good diagnostic image quality.