1.Detect of P53,bax and caspase 3 genes expression after optic nerve injury in rats with SYBR green I fluorescence quantitative PCR
Yingjuan LÜ ; Xiulan, ZHAO ; Jie, YANG ; Jinguo, YU ; Hua, YAN
Chinese Ophthalmic Research 2009;27(11):973-977
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.
2.Establishment of animal model of traumatic optic neuropathy and its function evaluation by F-VEP
Rongguo, YU ; Jinguo, YU ; Zhansheng SHEN ; Xing, WANG ; Yingjuan LÜ ; Wei, ZHANG ; Hua, YAN
Chinese Ophthalmic Research 2010;28(1):39-43
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.
3.Norm construction of Sub-Health Measurement Scale for nurses in Shandong Province
Xiaoyan LÜ ; Yingjuan CAO ; Shaowei SANG ; Rong LI ; Li LI ; Fuzhong XUE ; Jing DU
Chinese Journal of Nursing 2023;58(21):2637-2644
Objective To establish the norm of Sub-Health Measurement Scale for nurses in Shandong Province,so as to provide a basis for understanding the current situation of sub-health and provide a reference for promoting the health of nurses.Methods From December 2020 to February 2023,14,308 nurses from 23 public tertiary hospitals enrolled in the Nurse Health Cohort Study in Shandong Province were selected as participants by convenience sampling.The sub-health status was assessed by the Sub-Health Measurement Scale,and the sub-health norm of nurses was constructed by establishing the mean,percentile and demarcation norm.Results A total of 10669 nurses were included in the analysis.The total score of Sub-Health Measurement Scale of nurses in Shandong Province was(61.13±12.40),and the scores of physiological sub-health sub-scale,psychological sub-health sub-scale and social sub-health sub-scale were(58.35±11.86),(62.56±15.85)and(63.56±16.73),respectively.The mean norm of sub-health assessment scale was established by gender and age.Based on the results of univariate analysis,we selected the characteristics with differences,namely gender,age,marital status,working department,years of working,professional title and employment type to establish a classification norm.P5,P25,P50,P75 and P95 were select to establish percentile norm.Taking(x-s),(x-0.5s),(x+0.5s),(x+s)as the cut-off points,the demarcation norm of the Sub-Health Measurement Scale and sub-scales was established,which was divided into 5 categories,including disease,severe sub-health,moderate sub-health,mild sub-health and healthy states.Conclusion This study established the Sub-Health Measurement Scale norm of nurses,which provided a basis for effectively evaluating and identifying the sub-health status of nurses.