1.Fas gene expression of intervertebral disc in the patients with intervertebral disc herniation
Haoran Lü ; Jinshun YANG ; Yan HUANG ; Yu ZHAO ; Shanwei FENG
Chinese Journal of Tissue Engineering Research 2013;(43):7587-7593
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.
2.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.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.
4.Progress on Hypoxic-ischemic Brain Damage Associated with CCR2 and CCL2.
Yu-jia LUO ; Ru-bo LI ; Shi-yu MA ; Meng-yan LÜ
Journal of Forensic Medicine 2016;32(1):54-57
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*
5.Therapeutic effect of Hongbeiyegen on alcohol-induced rat hepatic fibrosis.
Xiao-Yan LÜ ; Qiang LIU ; Yu-Yao CHEN ; Yu-Hong SONG ; Zhi-Ping LÜ
Journal of Southern Medical University 2007;27(2):153-155
OBJECTIVETo observe the therapeutic effect of Hongbeiyegen [the root of Alchornea trewioides(Benth.) Muell.-Arg.] on alcohol-induced liver fibrosis (AF) in rats and explore its mechanism.
METHODSIn rats with AF, the serum levels of transforming growth factor beta1 (TGFbeta1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were detected along with examination of the changes in serum hyaluronic acid (HA), laminin (LN), procolagen type III (PC III), collagen type IV (C IV), glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) levels.
RESULTSCompared with the control group, Hongbeiyegen could significantly reduce the levels of TGFbeta1, TIMP-1, HA, LN, PC III, CIV, ALT and AST in rats with AF.
CONCLUSIONHongbeiyegen can relieve and ameliorate liver fibrosis possibly by inhibiting the expression of TGFbeta1 and TIMP-1.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Collagen Type III ; blood ; Collagen Type IV ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Ethanol ; Euphorbiaceae ; chemistry ; Female ; Hyaluronic Acid ; blood ; Laminin ; blood ; Liver Cirrhosis, Experimental ; blood ; chemically induced ; drug therapy ; Male ; Phytotherapy ; Plant Roots ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; blood ; Transforming Growth Factor beta1 ; blood
6.Establishment and identification of stable cell line secreting humen IgE Cε-4 protein
Chunxia QIAO ; Leiming GUO ; Ming Lü ; Ming YU ; Yan LI ; Jiannan FENG ; Beifen SHEN
Chinese Journal of Microbiology and Immunology 2008;28(7):661-665
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.
7.Evaluation of myocardial viability in myocardial infarction by low kilovoltage contrast-enhanced multi-slice CT
Qiang FAN ; Zhaoqi ZHANG ; Zhanming FAN ; Biao Lü ; Wei YU ; Zixu YAN ; Yike ZHAO
Chinese Journal of Radiology 2008;42(10):1040-1044
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.
8.Low-dose CT coronary angiography with prospective electrocardiogram-triggering using dual-source CT scanner
Lei XU ; Zixu YAN ; Zhaoqi ZHANG ; Zhanming FAN ; Biao Lü ; Wei YU
Chinese Journal of Radiology 2009;43(7):700-703
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.
9.A comparative study on continuous hemihepatic with intermittent total hepatic inflow occlusion in hepatectomy for liver tumors
Zhixue LIAO ; Tianfu WEN ; Zheyu CHEN ; Lunan YAN ; Jian YANG ; Bo Lü ; Guochang WU ; Yu ZHANG
Chinese Journal of General Surgery 2009;24(4):295-299
Objective To evaluate if continuous hemihepatic inflow occlusion(HH)during hepatectomy can be as safe and effective as intermittent total hepatic inflow occlusion(TH)in reducing blood loss during hepatectomy.Methods From November 2001 to March 2006.eighty patients undergoing liver resections were included in a prospective randomized study comparning the intra-and postoperative course underTH(n=40)or HH(n=40).TH was performed with periods of 20 minutes of occlusion and 5 minutes of releasing,while HH with continuous occlusion.The surface area of liver transection was measured and blood loss was calculated.The amount of blood loss,levels of alanine aminotransferuse (ALT)and aspartate aminotransferase(AST),and postoperative course were recorded. Results The total ischemic time of the HH groups was longer than in the TH group[(42±13)min,(31±13)min,P=0.37],and the operative time in the HH group was longer than in the TH group[(236 ±49)min,(204±38)min,P=0.02 ].No signincant difierenee was found between HH and TH group in blood loss during liver parenchyma transection[(500 ±269)ml,(416 ±235)ml,P=0.14]and in the changes of ALT and AST on the first postoperative day[ALT:(677±572)IU/L,(577 ±327)IU/L,P=0.12;AST:(591 ±468)IU/L,(512±301)IU/L,P=0.66].There were no difierences on postoperative morbidity between the two groups(22.5%versus 20.0%,P=0.35).Conclusion The technique of continuous hemihepatic inflow occlusion is as safe and effective as intermittent total hepatic inflow occlusion.
10.Diagnosis and treatment of cerebrospinal fluid leakage
Yong YAN ; Lijun HOU ; Mingkun YU ; Juxiang CHEN ; Liquan Lü ; Xiangqian QI ; Yicheng LU
Chinese Journal of Trauma 2009;25(1):32-34
Objective To investigate standard diagnosis and treatment of cerebrospinal fluid (CSF) leakage to improve the prognosis of the patients. Methods A retrospective study was done on 75 patients with CSF leakage from January 2004 to March 2007 in our hospital. There were 51 patients with rhinorrhea, nine with otorrhea and 15 with wound/incision leakage. Of all, 39 patients had traumatic leakage, 32 postoperative leakage and four spontaneous leakage. In the study, 23 patients were cured by position testing and drug therapy and 16 by cerebrospinal fluid drainage and/or wound debridement but 36 were treated with surgeries including craniotomy repair in 17, extracranial repair in 17 and CSF shunt in five (three received CSF shunt after repair). Results Of all, 64 patients were cured, 10 gained im-provement but one died. Conclusion Standard diagnosis and treatment of CSF leakage helps improve cure rate and reduce complications.