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.
3.Clinical observation on the reconstruction of large areas lower eyelid defect with Medpor spacer graft
Hai-Yan, LI ; Juan, SHAO ; Bo-Chang, LÜ ; Hua, ZHANG ; Bing-Yu, TIAN
International Eye Science 2014;(8):1537-1539
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.
4.Study on macular retinal thickness in young people by using 3 D optical coherence tomography
Gui-Ling, ZHAO ; Yan-Hua, PANG ; De-Mao, LIANG ; Qiu-Rong, LÜ
International Eye Science 2014;(8):1450-1453
AIM: To measure the macular thickness of normal young people by 3D 1000 optical coherence tomography (OCT) and study the repeatability of measuring results and the relationship between the thicknesses of macular and gender. At the same time, to compare our result with the data of other types of OCT, and to understand the consistency of the measuring results of macular thickness of different types of OCT.
METHODS: Totally 222 eyes in 111 young people were detected using 3D scan mode of Topcon 3D OCT 1000 (ver 2.4 ) . Twelve cases ( 24 eyes ) underwent repeatability check. We took transverse comparison between our measured results with other research's results.
RESULTS: There were 111 cases of young people, whose age were from 18-27 years old, all uncorrected and corrected visual acuity were≥1. 0, all intraocular pressure were <21mmHg. The average thickness of all macular region was 273. 32±17.08μm. Retinal volume of macular area was 7. 73 ± 0.37mm3 . Center thickness was 161 -264μm, and the average thickness was 200. 13±18. 81μm. Central macular thickness were 188 - 273μm, and the average thickness was 229. 00 ± 18. 20μm. The central macular thickness in men was significantly greater than that in women, and there was statistical difference. The results of repeated check of 12 cases ( 24 eyes ) in the macular area were no statistical difference except the outer ring of nasal quadrant, and the repeatability of average thickness in central macular thickness was better than in center thickness.
CONCLUSION:The repeatability of macular examination is good. The central macular thickness can be better repeated than the center thickness. The central macular thickness is 229. 00±18. 20μm in young people, according to the 3D 1000 OCT measurements. There are statistical difference of central macular thickness between different genders.
5.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.
6.XBP-1 interacts with estrogen receptor alpha (ERalpha).
Li-Hua DING ; Qi-Nong YE ; Jing-Hua YAN ; Jian-Hua ZHU ; Qiu-Jun LÜ ; Zong-Hua WANG ; Cui-Fen HUANG
Chinese Journal of Biotechnology 2004;20(3):332-336
Estrogen receptor alpha (ERalpha) has been a primary target of treatment as well as a prognostic indicator for breast cancer. The level of human X-box binding protein 1 (XBP-1) mRNA was related with that of ERalpha in breast tumors and was over-expressed in some breast tumors. These previous studies suggested that XBP-1 may interact with ERalpha. XBP-1 has two isoforms, XBP-1S and XBP-1U, as the result of unique splicing. GST pull-down assay showed that both XBP-1S and XBP-1U bound to ERalpha in vitro. The binding of XBP-1S to ERalpha was stronger than that of XBP-1U to ERalpha. Co-immunoprecipitation revealed that the binding was in a ligand-independent manner. XBP-1S and XBP-1U interacted with the region of ERalpha that contains a DNA-binding domain. The ERalpha-interacting regions on XBP-1S and XBP-1U have been mapped to two regions, the N-terminal basic region leucine zipper domain (bzip) and the C-terminal activation domain. These findings suggest that XBP-1S and XBP-1U may participate in ERalpha signaling pathway through the mediation of ERalpha.
Breast Neoplasms
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genetics
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metabolism
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Cell Line, Tumor
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DNA-Binding Proteins
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genetics
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metabolism
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Estrogen Receptor alpha
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genetics
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metabolism
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Female
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Humans
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Protein Interaction Domains and Motifs
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physiology
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RNA, Messenger
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biosynthesis
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genetics
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Regulatory Factor X Transcription Factors
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Signal Transduction
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Transcription Factors
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genetics
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metabolism
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X-Box Binding Protein 1
7.Hybrid procedure in treating infants and children with pulmonary stenosis under echocardiographic guidance
Ye LU ; Xiuzhang Lü ; Jianpeng WANG ; Zhenhui ZHU ; Yongqing LI ; Kunjing PANG ; Shoujun LI ; Jun YAN ; Zhongdong HUA
Chinese Journal of Ultrasonography 2011;20(7):575-577
Objective To evaluate the results of hybrid procedure in treating infants and children with severe pulmonary stenosis (PS) and the clinic value of echocardiography in the whole procedure.Methods Hybrid therapy was performed in 43 patients [(7.1±6.8)months,ranging 5 days to 33 months] with severe PS.The echocardiography data in the whole procedure and follow up were reviewed.Results Aided by echocardiography,the hybrid therapy was successful in 43 cases without major complication.Immediately following valvuloplasty,the gradient across pulmonary valve decreased from (92.5±21.4)mm Hg to (23.6±13.0)mm Hg (P<0.05).The growth and development of 27 patients in follow-up study was well.No restenosis was found and tricuspid regurgitation decreased differently.Conclusions Hybrid procedure is effective and safe for the treatment of infants and children with severe PS.And echocardiography play an important role in the whole procedure.
8.Association between p53 gene codon 72 polymorphism and keloid in Chinese population.
Li YAN ; Xiao-yan LÜ ; Chun-mei WANG ; Rui CAO ; Yan-hua YIN ; Chun-shi JIA ; Qiang ZHUANG
Chinese Journal of Plastic Surgery 2007;23(5):428-430
OBJECTIVETo investigate the relationship between p53 gene codon 72 polymorphism and genetic predisposition to keloid in Chinese population.
METHODSPCR-based restriction fragment length polymorphism (PCR-RFLP) analysis was used to detect p53 gene codon 72 genotypes of 60 keloid samples and 102 whole blood samples from healthy controls in China.
RESULTSThere was no significant difference in the distribution of p53 gene codon 72 polymorphism between the keloid patients and the healthy controls (X2 = 2.910, P = 0.233), nor did the frequencies for Pro and Arg alleles (X2 = 0.882, P = 0.348), and there was no significant difference in the distribution of p53 gene codon 72 polymorphism in keloid patients and normal controls from China and Japan respectively (X2 = 3.942, P = 0.139; X2 = 3.260, P = 0.196). But the Arg/Arg genotype was significantly higher than the Pro/Pro genotype among the patients with keloid in shoulder and back (P < 0.01).
CONCLUSIONSThere was no significant association between the distribution of p53 gene codon 72 polymorphism and keloid in Chinese population, but Arg/Arg genotype may affect the formation of keloids in shoulder and back compared to others. Further research should be done to investigate the relationship between p53 gene codon 72 polymorphism and keloids in different sites.
Asian Continental Ancestry Group ; genetics ; Codon ; genetics ; Female ; Humans ; Keloid ; genetics ; Male ; Polymorphism, Restriction Fragment Length ; Tumor Suppressor Protein p53 ; genetics
9.Characteristics of uterine contraction and stages of labor under continuous epidural block anesthesia.
Chun-yan YIN ; Jing-zhen ZHOU ; Xiao-yan LÜ ; Xiao-peng HUANG ; Gui-hua HE ; Jian-fei CHEN
Journal of Southern Medical University 2006;26(11):1563-1567
OBJECTIVETo observe the characteristics of uterine contraction and stages of labor during delivery under continuous epidural block anesthesia.
METHODSTotaling 213 parturients in spontaneous labor under epidural block anesthesia with dilated cervical orifice of 3 cm were monitored for the contraction cycle, duration, intensity and curve types of uterine contraction, and recordings were made for 30 min before and 30, 60 and 120 min after the anesthesia took effect, respectively. The duration of the active phase in the first, second and third stages of labor was compared between 421 cases with anesthesia and 237 without anesthesia.
RESULTSSignificant difference was noted in the objective indexes of uterine contraction recorded after anesthesia had taken effect (P<0.05) in comparison with those before anesthesia, suggesting significantly attenuated uterine contraction after anesthesia, whereas these indexes underwent no significant further variation as compared between different time points after anesthesia (P>0.05). The average active phase in the first stage was significantly shorter in anesthesia group than that in the control group (P<0.05), but the average duration of the second and third stages of labor differed little between the two groups with appropriate use of oxytocin under strict monitoring (P>0.05). The rates of obstetric forceps utilization and use of oxytocin were higher in anesthesia group than in the control group (P<0.05).
CONCLUSIONEpidural block anesthesia produces certain influences on uterine contraction and stages of labor during delivery, for which appropriate treatment measures may prove beneficial.
Adult ; Anesthesia, Epidural ; methods ; Anesthesia, Obstetrical ; methods ; Female ; Humans ; Labor, Obstetric ; physiology ; Pregnancy ; Time Factors ; Uterine Contraction ; drug effects ; Uterus ; drug effects ; physiology
10.Experimental studies on the treatment of colon cancer by cytosine deaminase gene and 5-fluorocytosine.
Jian-Hua HUANG ; Yao WANG ; Chong-Hui LI ; Da-Guang ZHONG ; Hai-Yan LÜ
Chinese Journal of Oncology 2005;27(1):6-8
OBJECTIVETo investigate the effect of cytosine deaminase (CD) gene plus 5-fluorocytosine (5-Fc) on the growth of human colon cancer xenograftin nude mice.
METHODSRetroviral vector expressing CD gene was transfected into human colon cancer SW1116 cells. Expression of the transfected CD gene in SW1116 (SWCD(2)) was confirmed by RT-PCR. The cytotoxicity of 5-Fc on SW1116 was determined by MTT assay in vitro. In vivo, the CD gene expression vector was injected intratumorally and 5-Fc was given by ip injections.
RESULTSIn vitro, SWCD(2) cells were killed by 5-Fc with an IC(50) of 66 micromol/L while the nontrasfected SW1116 cells needed an IC(50) of 16 000 micromol/L to be killed. The growth of SWCD(2) xenografts was significantly inhibited by systemic administration of 5-Fc.
CONCLUSIONCD gene/5-Fc system is a potential gene therapy strategy for human colon cancer.
Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; metabolism ; pathology ; prevention & control ; Combined Modality Therapy ; Cytosine Deaminase ; genetics ; metabolism ; Female ; Flucytosine ; pharmacology ; therapeutic use ; Genetic Therapy ; Genetic Vectors ; Humans ; Inhibitory Concentration 50 ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Retroviridae ; genetics ; Transfection