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Table of Content

    25 June 2007, Volume 27 Issue 6
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    研究论文
    a-synuclein phosphorylation increased TH activity in MN9D cells
    2007, 27(6):  601-604. 
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    Objective To observe the effect of phosphorylated α-Syn on TH activity. Methods pLNCX2-S129D α-Syn vector was constructed using overlapping extension polymerase chain reaction (OE-PCR). The retroviral particles were packaged and infected MN9D cells. Expression of α-Syn in MN9D was tested using real time RT-PCR. Phosphorylated TH protein level was tested using Western blot. Results Sequences of wild-type and S129D α-Syn were both correct. The result of real time RT-PCR showed that wild-type and S129D α-Syn genes were overexpressed in MN9D compared with uninfected MN9D and vector control group. TH phosphorylation was reduced in wild-type α-Syn group, while increased significantly in S129D α-Syn group. Conclusion　α-Syn phosphorylation involves in regulation of TH activity in MN9D cells.
    Expression changes of Ephrin B1 and Ryk in DRG after peripheral nerve injury
    2007, 27(6):  605-609. 
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    Abstract: Objective To study expression changes of Ephrin and its receptors in DRG neurons after peripheral nerve injury. Methods Set up animal model of unilateral crushed sciatic nerve, and by immunostaining screened the expression pattern changes of Ephrin B1, Eph B1、Eph B2、Eph B3、EphA4 and Ryk in DRG neurons after unilateral sciatic nerve crushed, quantified the positive staining cells in DRG and analyzed the percentage of positive cells in different size. Results After peripheral nerve injury, the expression of Ephrin B1 significantly decrease, at the same time there were no any expression change of Eph B1、Eph B2、Eph B3 or Eph A4, but the Ryk expression was intensely induced in DRG neurons. Conclusion Because the Ephrin B1 and Ryk show very strong expression changes in DRG neurons after peripheral nerve injury, so this result strongly suggests they might involved in the process of peripheral never regeneration.
    Effect of ischemic pretreatment on levels of cPKC isoform-specific membrane translocation and protein expression in retina of rats
    2007, 27(6):  610-615. 
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    Objective To identify certain isoforms involved in the onset of retinal ischemic preconditioning (IPC), the effects of ischemic pretreatment (IP) were observed on levels of conventional protein kinase C (cPKC) a, bI, bII and g isoform-specific membrane translocation and protein expression in retina of rats. Methods Retinal IP was produced by intra-ocular pressure (IOP) elevation for 5 minutes in anesthetized Wistar rats. Sham operation was similar to IP except the pressure elevation. 10, 20, or 40 minutes and 1, 12, 24, 72, or 168 hours after the procedure, levels of cPKC isoform-specific membrane translocation and protein expression were analyzed using Western-blot. Results cPKCa protein expression levels increased significantly from 12h to 168h. A peak reached at 72h after IP. cPKCg membrane translocation enhanced during 20min to 1h with a peak at 40min. cPKCg protein expression levels increased significantly from 12h to 72h. A peak was at 24h after IP. However, there were no significant changes in both membrane translocation of cPKCa, bI, bII and protein expression of cPKCbI, bII in retina of rats following IP. Conclusion The enhanced cPKCg membrane translocation, and the increased cPKCa and g protein expressions might be involved in the onset and sustain of retinal IPC in rats respectively.
    Effect of mCD99L2 gene on the cell biological characteristic of mice lymphoma cell A20
    2007, 27(6):  616-620. 
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    Objective To investigate the effects of mCD99L2 gene silencing on the mice B lymphoma cell line A20 transferred to H/RS-like cells. Method Recombinant plasmid LV-mCD99L2 was transfected into A20 cells which endogenetically express mCD99L2. The stably expressed cell line was selected by Blasticidin. The expression of CD30 in transferred cells was detected by immunofluorescense and FCM method. Observation of A20-LV-mCD99L2 cells was applied by transmission electron microscope. The proliferation of the tumor cells (the diameter of H/RS-like ≥25μm) of A20- LV- mCD99L2 and A20 in vitro and in nude mice was examined by continuously cell counting, and taken the human Hodgkin lymphoma cell line L428 as control. The cell cycle was detected by FCM method. Result Stable expression cell clone A20-LV- mCD99L2 was obtained after recombinant plasmid transfection. The expression of CD30 in transferred cells was positive, and the rate of expression of CD30 in A20-LV- mCD99L2 was 54.4%. The observation of transferred cells showed that the cells nuclear were larger and H/RS-like cells with obvious nucleoli which were uninucleated, binucleated and multinucleated cells. The result of cell counting showed that the rate of H/RS-like cells of A20-LV-mCD99L2 was obviously higher than that of A20（P<0.01）in vitro and in vivo. Cell cycle analysis showed that no obvious differences between A20-LV- mCD99L2 and A20 in S stage and no obvious apoptosis was detected in A20-LV- mCD99L2 and A20. Conclusion mCD99L2 gene silencing may transfer mice B lymphoma cell line A20 into H/RS-like cells in vitro and in vivo.
    The protective and therapeutic effects of adrenomedullin on hypoxia pulmonary hypertension in rats
    2007, 27(6):  621-624. 
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    Objective To investigate the effects and mechanisms of adrenomedullin (ADM) and adrenomedullin receptor (ADMR) on hypoxia pulmonary hypertension (HPH) in rats.Method Eighteen male Wistar rats were randomly divided into three groups:control group and hypoxic group, hypoxic with ADM group .ADM was subcutaneously administered into rats with ADM by osmotic minipump（1.5μg·kg-1·h-1）.After 21d ,The Mean pulmonary arterial pressure (mPAP),right ventricular hypertrophy index(RV/(LV+S)) were measured and hypoxic pulmonary vascular remodeling(HPSR) were observed with morphometric analysis. The levels of pulmonary arterial plasma ADM were measured by radioimmunoassay. The semiquantitate in situ hybridization was used to measure the expression of ADMR mRNA in pulmonary artery walls. Result (1) The level of mPAP, RV/(LV+S) ,the ratio of vascular wall thick ness to external diameter(MT%) and ratio of vascular wall area to total area (MA%) were significantly higher on hypoxia group than those in (2) The pulmonary artery plasma ADM level was significantly higher in hypoxic group and hypoxic with ADM group than that in control group (P<0.01), and the ADM level decreased in rat of hypoxic with ADM group compared with hypoxic group(P<0.05).(3) ADMR mRNA expressed higher in pulmonary artery walls of hypoxia group and hypoxic with ADM group（P<0.01）.Conclusion ADM subcutaneously administered may partly prevent the development of HPH and pulmonary vascular remodling. Exogenous ADM had no negative feedback on pulmonary arterial plasma ADM and ADMR mRNA.
    Relationship between inflammatory factor and artery dilation
    2007, 27(6):  625-628. 
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    Objective To investigate the relationship between inflammatory factors and the coronary artery dilation, and the clinical significance of their changes. Methods The cases performed coronary angiography in our hospital last year was collected, and they were divided into three groups , the first one included 11 patients whose angiography were coronary artery dilation, the second group included 35 cases whose results were atherosclerosis, and the third was normal angiography which included 24 cases. sES, MMP9 and TIMP1 were measured by ELISA method, and the results are compared. Results Patients with coronary artery dilation were found to have significantly higher in comparison with atherosclerosis group and normal group(153.7±152.7ng/L，90.1±54.2ng/L，76.5±37.2ng/L, respectively)(P value=0.039)，there were also statistic significance of MMP9 among three groups(2184±1723ng/L， 949± 827ng/L，1274± 1394ng/L）（P＝0.016）, but there were no statistic significant difference of TIMP-1 among three groups(8106±1218ng/L，8203±834ng/L，8433±867ng/L）（P>0.05）. Conclusion sES and MMP-9 are associated with coronary artery dilation , they maybe attribute to the coronary artery dilation, and they might play important roles in the coronary artery pathogenesis.
    Analysis of changes in TCRBV and CDR3 diversity before and after auto-PBSCT for severe/refractory CTD
    2007, 27(6):  629-634. 
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    Objectives To study the changes of clonality of T cell receptor (TCR) and complementarity determinative region 3(CDR3) before and after autologous peripheral blood CD34+ stem cell transplantation(auto-PBSCT) for severe/refractory connective tissue disease（CTD）. Methods Thirteen patients with severe/refractory CTD were enrolled for auto-PBSCT in Peking Union Medical College Hospital, including systemic lupus erythematosus (SLE) 8 cases, rheumatoid arthritis(RA) 4 cases, and primary Sjogren's Syndrome( pSS) 1 case. Blood samples were collected before/after mobilization, 2 weeks, 1 month, 3 months , 6 months 12 months 18 months post-transplantation. Diversity of TCR BV and CDR3 were showed by reverse transcription- polymerase chain reaction( RT-PCR) and genescan. Rusualts The TCR BV usage and CDR3 spectral pattern of pre-auto-PBSCT CTD patients were revealed skewed pattern and oligoclonality, Which developed severe oligoclonality within 1 months after auto-PBSCT. However, they showed diversity and poly-clonality 3-6 months after auto-PBSCT. Conclusion Skewed pattern and oligoclonality of TCR BV and CDR3 which implied auto-reactive were depressed after auto-PBSCT, and inclined to change to normal pattern.
    The Relationship Between Lateral Hypothalamus Area (Lha) And Small Intestine Moving
    2007, 27(6):  648-651. 
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    The present research discussed the influence of Noradrenergic systems in Lateral Hypothalamus Area (LHA) on small intestine moving and possible operating mechanism.. Methods: The effect of injecting different drugs including saline、 noradrenalin、noradrenalin+phentolamine、noradrenalin+propolol、phentolamine on small intestines electro-activity of rats was detected by external alimentary canal electrodes, and by the central nervous system stereo-configuration technology, in which the Migrating Myoelectric Complex (MMC) indexes as the cyclic period, the ratio of the active time to the cyclic period, and the number of the fast wave within the active time per minute were used..Results: For LHA, the cyclic period of MMC extended, the ratio of the active time to the cyclic period and the number of the fast wave within the active time per minute reduced, after injecting 4mg/2ml Noradrenaline (NE). These results were considerably different with the saline group. NE inhibited the electro-activity of small intestine of rats, and the average effective time was 36.86+7.39 min. This function was retarded by injecting 10mg/2ml phentolamine into LHA, rather than the same dose of propolol. Hence injecting 10mg/2ml PE alone to LHA raised the ratio of the MMC active time to the cyclic period, which obviously appeared a disagreement compared with the saline group. And phentolamine presented a slight excitability on the electro-activity of small intestine of rats. Conclusion: The NE in LHA showed the inhibitive regulation effect on small intestines, and this effect was introduced through a acceptor and might be irrelevant to b acceptor. The a acceptor in LHA involved in the inhibition of the electro-activity in small intestines.
    技术与方法
    Confiramtion of Animal Model of Cognitive Impairment Induced by Cerebral Ischemia in Diabetic Rats
    2007, 27(6):  680-684. 
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    Objective: The aim of this study was to prepare a new model of cognitive impairment induced by cerebral ischemia in diabetic rats. Method: A total of 70 female Wistar rats，weighing 150-200g (around 6 weeks of age), were randomly divided into 5 groups: a normal control group (n=15), an ischemic group(n=18), a diabetic sham-operation group (n=19) and a diabetic ischemic group (n=18). To induce diabetes, streptozotocin (STZ) was injected i.p. at a dose of 60 mg/kg, 4 days before the preparation of experimental ischemia. The method used to produce cerebral ischemia was the two-vessel occlusion followed by re-perfusion. Under anesthesia with sodium pentobarbital, both common carotid arteries were exposed through a ventral midline cervical incision. The common carotid arteries were occluded with an artery forceps for 15min which was followed by a re-perfusion for 5min. The operation of occlusion and re-perfusion was performed twice. The rats for sham-operation controls (operated at the same time points) were treated in the same manner except the occlusion of common carotid arteries. The blood sugar level was determined 7 days after STZ administration. The animals with the blood glucose concentration higher than 15.5mM were selected as diabetic rats. After 1-month recovery the animal model with cognitive impairment was confirmed by behavioral detection. The animals Learning and memorial behaviors were investigated using a passive and active avoidance response tests and a spatial version of the Morris water maze test. The situation of neuron loss in the area of hippocampus was determined by the pathological H-E staining method. Result: The performance of passive avoidance in diabetic rats with cerebral ischemia was significantly impaired. The step-down latency of the passive avoidance response of the model rats was shorter, especially at 5th min after training (P<0.01), than that in other groups 30 days after production of cerebral ischemia. During 5 days-training of active avoidance, the value of AR in the model group was less than that of the control group, especially at the 1st and 3rd day, there was statistical significance between the two groups (P<0.01). The training times of the model group were significantly more than that of the other three groups (P<0.001). The swimming time of the rats in Morris water maze in model group was significantly shorter than that in other groups (P<0.05 as compared with DM+Sham; P<0.01 as compared with other 2 groups). The swimming distance of rats in model group was the shortest among all 4 groups (P<0.05). The result indicated that the memory ability in the model rats was deficient. Conclusion: The cognitive ability in diabetic rats with cerebral ischemia 30 days after being induced was significantly declined compared with the other three groups and diabetes can worsen neuronal damage of the brain via cerebral ischemia.
    Study of perfused porcine anterior segment and tissue sheet cultured porcine trabecular meshwork
    2007, 27(6):  690-692. 
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    Objective To study the differences of trabecular meshwork cells cultured by perfused anterior segment and tissue sheet culture method, and to clarify their application in glaucoma study. Methods Trabecular meshwork cells were cultured by perfused anterior segment and tissue sheet culture method, light microscope was used to observe the trabecular meshwork cells and intercellular spaces. Results IOP of the porcine anterior segments perfused under constant rate at 0.1ml/h could maintain in the normal range（10~12mmHg）. The success rate was 50%. The IOP could be elevated with the increasing of perfusion rate, while the shape and structure of the organ were well preserved. Trabecular meshwork cultured by tissue sheet culture method could also reserve the tissues well, but the intercelluar spaces collapsed . Conclusion Perfused anterior segment could better simulate the normal physical state in situ and adequate perfusion was necessary for normal trabecular meshwork.
    临床园地
    研究短文
    短篇综述
    医学教育
    The Discussion Based On Cases ---- A Binding Of Physiology and Clinic
    2007, 27(6):  719-720. 
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    To study how to do the discussion class based on cases in the physiological teaching in this paper. Firstly the teacher should carefully select typical cases. Secondly the teacher should assure the discussion encircle physiology. At the same time, the teacher should try to create a good atmosphere and encourage the student to discuss question. Finally it is important to collect feedback information.
Basic & Clinical Medicine
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Add: 5 Dong Dan San Tiao, Beijing 100005
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E-mail: basic_clinic@vip.163.com
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