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Table of Content

    25 March 2009, Volume 29 Issue 3
    研究论文
    Clinical and Molecular Genetic Analysis on a Patient with 17 Hydroxylase /17, 20 Lyase Deficiency
    Cai-xia CAO; Ou WANG; Min NIE; Yu-xiu LI; An-li TONG; Lin LU; Zhao-lin LU
    2009, 29(3):  225-228. 
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    Objective  To analyze the clinical and molecular genetic characteristics of a Chinese patient with 17α- hydroxylase/17, 20 Lyase deficiency(17OHD). Methods Clinical features and laboratory data were collected . Genomic DNA was extracted from leukocytes of peripheral blood of the patient. All eight exons of the CYP17 gene, including the flanking regions of introns, were amplified by PCR. The mutations of the CYP17A1 gene were analyzed by direct sequencing the amplified DNA fragments. Results The patient was diagnosed as 17OHD according to the clinical presentations , laboratory examinations and CYP17A1 mutation which was identified as a base deletion and a base transversion ( TAC/AA) at codon 329 and caused a missense mutation of Tyr→Lys at this codon and the open reading frame shift following this codon to produce a truncated enzyme without activity site. Conclusions Through CYP17A1 gene mutation analysis, the clinical diagnosis of 17OHD was confirmed . The alternation of P450c17 structure caused by CYP17A1 gene mutation is the molecular mechanisms of clinical manifestation of the patient.
    Fusion expression of peptide with high affinity to human breast cancer and its targeting transportation effect
    Wei-qing LIU; Jian DONG; De-ping ZHAO; Min HONG; Jun YANG; Hai-xia XING
    2009, 29(3):  229-233. 
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    Objective To investigate targeting delivery of a peptide with high affinity to human breast cancer in different types of human breast cancer cells. Methods Gene of peptide was clone into pGEX-2T vector. The fusion proteins of glutathione-S-transferase (GST) and peptide sequence CASPSGALRSC (PⅠ), named GST-PⅠ, were expressed in BL21 (DE3) and purified by GST affinity column. The purified fusion protein was detected by SDS-PAGE and Western-blotting. Immunostaining analysis was used to determine the transduction of GST-PⅠ fusion protein into the MDA-MB-231 cells and other three human breast cancer cell lines were selected as control. Results A series of pGEX-2T-PⅠ vectors were constructed. The result of Western-blotting showed that the fusion protein was well immunoreacted with the mouse anti Schistosom a japonica GST monoclonal antibody. Immunostaining analysis showed that, contrasted to controls, GST-PⅠ could targetingly permeate into the MDA-MB-231 cells. Conclusions GST-PⅠ, a fusion protein with expected molecular mass, was expressed. PⅠ can mediate the fusion proteins across the cell membrane. It can be used as a targeting vector to delivery bimolecular agent in gene therapy cases of breast cancer.
    Protective effect of Akt on gastric ischemia-reperfusion injury by microinjection of oxytocin into the paraventricular nucleus in rats
    Yong-mei ZHANG; Wen-wen ZHANG; Jian-fu ZHANG
    2009, 29(3):  234-237. 
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    Objective To observe the effect of microinjection of oxytocin (OT) into paraventricular nucleus (PVN) on gastric ischemia-reperfusion (GI-R) injury and its local molecular mechanism. Methods GI-R injury was induced in rats by clamping the celiac artery for30 min and then reperfusing for 1 h. A cannula was inserted into the unilateral PVN for microinjection of OT. The gastric mucosal injury index was counted grossly. The expressions of Akt and caspase-3 in rat gastric mucosa were examined by Western blot and by immunohistochemistry. Results Microinjection of OT into PVN dose-dependently attenuated gastric mucosal injury subjected to GI-R. Microinjection OT into PVN significantly increased the expression of Akt protein and decreased the level of caspase-3 in gastric mucosal following GI-R. The effects of OT were prevented by pretreatment with OT receptor antagonist atosiban into the lateral cerebral ventricle. Conclusion Microinjection of OT into PVN significantly protected against GI-R injury. These central effects of OT are mediated by its receptors. The local mechanisms were mediated by increasing Akt expression, which in turn leads to inhibit caspase-3 expression.
    Effects of transplantation of MSCs on the key enzymes of energy metabolism in acute myocardial infarction rat
    Wang LI; Sheng-shou HU; Ying-jie WEI; Hao ZHANG; Jian-feng HOU; Bo YE
    2009, 29(3):  238-241. 
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    Objective To investigate the effects of transplantation of MSCs on the key enzymes of energy metabolism after acute myocardial infarction(AMI). Methods Eighteen SD rats were randomly divided into 3 groups(n=6 respectively):control group,acute myocardial infarction group,MSCs group. The left ventricular tissues of rats were isolated one day later after left anterior descending artery occlusion for determination of HK, CACT,ACS,CS,LDH by ELISA. Result One day post-AMI,the concentration and activity of these enzymes changed obviously(P<0.05~P<0.01). One day after MSCs treatment post-AMI,some of concentration and activity of those enzymes had returned to normal levels. Conclusion The transplantation of MSCs may improve cadiocyte's energetics by regulating the concentration and activity of those key enzymes .
    The effect of pulmonary neuroendocrine cells following bronchoplasty in rats
    Kun-xiang GAO; Yun-jie WANG; Zhi-pei ZHANG; Zheng-yuan ZHAO; Xiao-long YAN; Ying-chun DENG; Yong HAN
    2009, 29(3):  242-246. 
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    Objective To explore the role of pulmonary neuroendocrine cells(PNECs)in damage and repair of airway by the changes of amount, distribution of PNECs after bronchoplasty. Methods 30 sprague dawley rats were randomly divided into three groups according to sample taking time after left principal bronchoplasty:1 week, 1 month, 3 months, and another 10 rats without bronchoplasty were used as normal control. The quantity and distribution of PNECs were studied by Grimelius silver stain. Both serotonin (5-HT) positive PNECs and calcitonin gene-related peptide (CGRP) positive PNECs were studied by imnunohistochemistry. Results PNECs were mainly found in the epithelium of the bronchus and terminal bronchioles. Some PNECs formed into neuroepithlial bodies (NEB). 1 wk after bronchoplasty, the amount of PNECs was significantly increased, and gradually declined to the normal control after 3 months . Except their location in the larger bronchi, 5-HT positive PNECs and CGRP positive PNECs did the same in number and NEB formed. Conclusion The quantity of PNEC and 5-HT positive PNEC and CGRP positive PNEC were significantly increased after bronchoplasty. It deduced that PNEC might be taken part in damage and repair of airway.
    Induced Differentiation in vitro of Rat ADSCs into Photoceptors and RPE Cells
    Zhuo-zai XU; Fang-tian DONG; Lian-feng CHEN; Chan WU; Rong-ping DAI; Wei-hong YU
    2009, 29(3):  247-253. 
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    Objective To explore differentiation in-vitro of rat adipose-derived stem cells into photoreceptor cells and RPE cells.Methods: The ADSCs were cultured by adhering to the flask surface and purified by continual passage. Surface antigens including CD45、CD90、CD49d、CD106 were indentified by flow cytometry. ADSCs were induced to differentiate by EGF, activin A, taurine, retinoic acid(RA) and extracted liquid of retina respectively. Meanwhile, ADSCs were induced by EGF+taurine, EGF+RA, taurine + RA, EGF+taurine+RA respectively. Immunofluorescence was used for detecting the expression of rhodopsin, CK and S-100, and flow cytometry was used for quantification. Results: For primary culture,the phenotypes of ADSCs were: CD45, CD90, CD49d and CD106, with a positive percentage of 1.6%, 71.3%, 7.8% and 3.5%, respectively. From passage 1 to 5, these phenotypes were: CD45(0.8%~9.3%), CD90(84.7%~94.8%), CD49d(16.8%~31.0%)and CD106(8.3%~22.2%). There was a higher CD49d percentage than CD106 in all the passages. The induction efficacy of ADSCs was 17.5%~46.0% for rhodopsin, 19.7%~79.3% for CK and 27.3%~50.7% for S-100. Conclusion: It is suggested that ADSC has potential to differentiate into photoceptors and RPE cells as evidenced by the presence of the specific markers of photoceptors(rhodopsin) and RPE markers(CK and S-100).
    Changes of learning and memory and calmodulin dependent protein kinaseⅡexpression in hippocampal neurons of pentylenetetrazole-kindled rats
    Yu YIN; Wen-qing ZHAO; Wen-ling LI; Pei-yuan LV; Wei-bo LI; Chuan-dong LIANG; Yue-hong WANG
    2009, 29(3):  254-258. 
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    Objective To observe the effect of pentylenetetrazole(PTZ)-kindled epliepsy on rat's learning and memory and the expression of calmodulin dependent protein kinaseⅡ(CaMKⅡ)in hippocampal neurons. Methods The rats were injected intraperitoneally with PTZ to establish the kindled models. The changes of behavior were evaluated through Morris water maze and the expression of α-CaMKⅡ in hippocampal neurons was observed by immunohistochemistry technique. Results The Morris water maze test showed that the escape latency and the swimming distance in PTZ-kindled rats increased significantly (P<0.05). The number of platform crossings and the percentage of swimming time in platform quadrant in PTZ-kindled rats decreased obviously (P<0.05), and the efficiency of their search strategy was poor. It showed that the ability of learning and memory in PTZ-kindled epliepsy rats was obviously impaired, and the expression of α-CaMKⅡ in hippocampal CA1 and CA3 sector significantly decreased (P<0.05). Conclusions The impaired ability of learning and memory in the PTZ-kindled epliepsy rats might be associated with decreased expression of CaMKⅡ in hippocampal CA1 and CA3 sector.
    Effect of JNK inhibitor on apoptosis and cell-cycle arrest of the Human Esophageal Cancer Cell Line Eca-109 induced by the derivative of D-amine-glucose
    Zhan-rong QIANG; Jing WU; Guo-dong YANG; Juan LI; Yong-ning ZHOU; Ai-qin WANG; Qun-ji XUE
    2009, 29(3):  259-263. 
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    Objective: To explore the effect of SP600125, a specific c-jun NH2 terminal protein kinase (JNK) inhibitor, on apoptosis and cell-cycle arrest of the human esophageal cancer cell line Eca-109 induced by 2-(3-carboxy-1-oxopropyl)amino-2-deoxy-D-Glucose (COPADG)and the possible molecular mechanism in COPADG-induced cell apoptosis was discussed. Methods: Eca-109 cells were cultured by using RPMI-1640 and calf serum. Eca-109 cells were pre-incubated with SP600125 for 30min prior to exposure to COPADG at different concentrations and for different time. Changes in expression of P-JNK protein were examined by Western blot;Cell growth inhibitory rate was detected by MTT colorimetric assay;Apoptosis rate and cell-cycle arrest was analyzed by flow cytometry. Results: COPADG could significantly inhibit the proliferation of Eca-109 cells and induce them to apoptosis and cell-cycle arrest in G0/G1 phase. Western blot showed that the protein expression of P-JNK was increased in a dose-dependent manner in Eca-109 cells after stimulated by COPADG. SP600125 remarkablely decreased the protein expression of P-JNK as well as the apoptosis rate and cell growth inhibitory rate in Eca-109 cells induced by COPADG as compared with those treated with only COPADG, meanwhile, cell-cycle arrest in G0/G1 phase progressed to cell-cycle arrest in G2/M phase. .Conclusion: JNK signaling pathway may play an important role in apoptosis of Eca-109 induced by the COPADG.
    Effect of BK Channels on Intracellular Free Calcium and Action Potential of Murine Cortical Neurons
    Wen-hua YANG; Xiang-jun BAI; Hong ZHAO
    2009, 29(3):  264-267. 
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    Objective To investigate the role of large conductance calcium-activited potassium channels(BK) in the regulation of intracelullar free calcium concentration([Ca2+]i) and. excitability of mouse cortical neuron Methods Culture neuronal cells of C57BL/6 mouse cortex. Observe the influence of IBERIOTOXIN(100nmmol/L) on neuronal action potentials and [Ca2+]i with patch-clamp techniques. evoke prolonged depolarization of cultured neurons by 100pA electric current lasting 500ms, Measure the rate of firing and [Ca2+]i. Perfuse extracellular solution with KCl(20mmol/L), compare the changes of [Ca2+]i with or without IBERIOTOXIN (100nmol/L). Results No significant changes of neuronal spontaneous action potential frequency and [Ca2+]i was observed in physiological condition after perfusion of IBERIOTOXIN (n=7,P>0.05). The frequency of action potential evoked by electric current is higher after perfusion of IBERIOTOXIN, as well as the [Ca2+]i level during depolarization(n=9,P<0.05). Extracellular high concentration of KCl increases [Ca2+]i of neurons. When IBERIOTOXIN was applied, [Ca2+]i was much higher than without IBERIOTOXIN (n=9,P<0.05). Conclusions BK may play an important part in the regulation of neuronal intracellular free calcium and excitability.
    Promotion of TRAIL-Induced Apoptosis by siRNA to FLIP gene in ovarian cancer cell line
    Yu WANG; Pei-shu LIU; Xiao-lei ZHANG; Cai-feng DAI; An-cong WANG; Juan LI
    2009, 29(3):  268-272. 
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    Objective To investigated the effect of FLIP-siRNA-mediated gene silencing on sensitivity of ovarian cancer cells to TRAIL. Methods Design and chemically synthesize three siRNAs based on the sequence of FLIP mRNA. They were transfected into ovarian cancer cell line A2780 with LipofectamineTM2000. The FLIP mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The FLIP-siRNA which has the most powerful inhibition was selected. Cell growth inhibition and apoptosis induced by TRAIL were observed with MTT assay and flow cytometry, respectively. Results Short siRNAs targeting FLIP down-regulated the mRNA and the protein level of FLIP oncogene in a time-dependent manner(P<0.01). Cell growth inhibition and apoptosis efficiency induced by TRAIL was obviously enhanced after FLIP-siRNA transfection(P<0.05). Conclusion FLIP-siRNAs can effectively inhibit FLIP expression in transcriptional and translational level, and greatly enhance TRAIL sensitivity in ovarian cancer cells.
    Marrow mesenchymal stem cells transplantation for ischemic heart disease in rabbits
    Guo-tao MA; Hua REN; Zhao-hui ZHU; Chao-ji ZHANG
    2009, 29(3):  273-276. 
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    Objective To investigate the feasibility of treating ischemic heart disease with transplantation of marrow mesenchymal stem cells. Methods Marrow mesenchymal stem cells were taken from New Zealand rabbit's sternum and cultured. A myocardial infarction model was created by ligation of the distal left anterior descending artery in New Zealand rabbit. MSCs were injected into the region of myocardial infarction. The size of the myocardial infarction area was measured by PET and the cardiac function was assessed by measuring the pressure change of left ventricle(dp/dt). Results The size of the myocardial infarction area diminished and the cardiac function of the New Zealand rabbit was improved after the transplantation of marrow mesenchymal stem cells. Conclusion Transplantation marrow mesenchymal stem cells may improve cardiac function of ischemic heart disease in New Zealand rabbit.
    Bone marrow mesenchymal stem cells from rats with hepatic fibrosis differentiate into hepatocyte-like cells in vitro
    Wen-yan HE; Jing LIU; Shu-xian LIU
    2009, 29(3):  277-282. 
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    Objective To investigate the difference of differentiation from rat bone marrow mesenchymal stem cells (MSCs) into hepatocyte-like cells from normal group and hepatic fibrosis model group. Methods Wistar rats were randomly divided into 2 groups: normal group and hepatic fibrosis model group. The liver fibrosis was induced by CCL4. MSCs were isolated by combining gradient density centrifugation with plastic adherence. Pure MSCs were obtained by cultivation and passage .The cells then treated with HGF and FGF-4. Levels of AFP and albumin in the supernatant were determined on day 15, 21 and 27. On day 27, cells of induced and non-induced were collected, glycogen store of hepatocytes was determined and the expressions of CK-18 and CK-19 were detected by immunocytochemical analysis. Results The levels of AFP in induced MSCs were higher on day 15, 21, 27, and reached to the highest on day 21; there was no significant difference between induced and non-induced MSCs in albumin levels on day 15, but on day 21, 27, compared with the non-induced MSCs, the albumin level in induced MSCs was higher and up to highest on day 27;glycogen storage of induced MSCs was observed on day 27 compared with non-induced MSCs; the induced MSCs expressed CK-18 and CK-19 while the non-induced MSCs did not. Compared by the levels of AFP and albumin, there was no significant difference in differentiation effect of MSCs between the normal group and hepatic fibrosis model group. Conclusions:Rat MSCs of hepatic fibrosis model group could differentiate into hepatocyte-like cells with hepatic phenotype and function in the presence HGF and FGF-4 in vitro.
    Effects of stromal cell-derived factor-1 on endothelial progenitor cells migration
    Yuan-bing WU; Yu-qi WANG; Wei-guo FU; Yang WANG
    2009, 29(3):  283-286. 
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    Objective To investigate the migration function of ex vivo expanded endothelial progenitor cell (EPC) by stromal cell-derived factor-1 (SDF-1). Methods Total mononuclear cells were isolated from human peripheral blood by density gradient centrifugation. Attached spindle -shaped cells were detached at day 7. The expression of CD34 and CD133 were determined by flow cytometry; KDR(VEGFR-2) was detected by reverse transcriptase-polymerase chain reaction; endothelial function was determined by cell absorption of DiI-acLDL and FITC-UEA-1; the expression of CXCR4 was determined by immunocytofluorescence. The migration function of EPC was determined using Transwell chamber. Results The positive rates of CD34, CD133 of EPC at 7 days of culture were 36.3%±23.8% and 19.7%±10.3% respectively. RT-PCR demonstrated the presence of KDR mRNA. EPC had the capacity to incorporate DiI-acLDL and FITC-UEA-1 ( >90% ). The expression of CXCR4 receptor was 74.8%. The migration cells of control, 10, 20 and 50μg/L were 3.5, 7.4, 24.9 and 28.0, respectively. The migration activity was significantly higher in groups of SDF-1 than that of control group (p<0.01). The difference was determined in comparison of 10μg/L vs 20μg/L (p<0.01), 20μg/L vs 50μg/L (p<0.05), and 10μg/Lvs 50μg/L (p<0.01), respectively. Conclusions 1. EPCs could be isolated and cultured from peripheral blood mononuclear cells. The CXCR4 receptor is present on EPC. 2. SDF-1 induces EPC migration in a dose-dependent manner.
    Application of WCX Magnetic Bead for Serum Proteome Profiling in Cervical Squamous Cell Carcinomas and Its Clinical Significance
    Zhi-guo ZHENG; Ting XIA; Yong-zhe LI; Yun GAO; Han-zhou MOU; Shen-hua XU; Yang XU
    2009, 29(3):  287-291. 
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    Objective To analyze the serum proteomic pattern of the cervical cancer patients,build diagnostic model and evaluate its clinic significance. Methods WCX magnetic bead and MALDI-TOF were used to detect the serum proteomic pattern of 77 patients with cervical squanmous cell carcinomas, 13 patients with CINⅢ and 52 healthy women. Biomarker Wizard software was used to detect protein peaks significantly different between cervical cancer and controls. The model was built by Biomarker Patterns software which was further valuated in blind test. Results A diagnostic pattern consisting of three differential protein peaks was established with 100%(32/32)sensitivity and 93.8%(30/32)specificity. A sensitivity of 77.8%(35/45)and a specificity of 75%(15/20)in blind test were obtained. The diagnostic model also could discriminate CINⅢ and SCC-Ag negative patients from controls. Conclusions The diagnostic pattern combining 3974,3398,13732m/z protein peaks can discriminate not only cervical squamous cell cancer but also CINⅢ and SCC-Ag negative patients from controls.
    SIGNIFICANCE OF UNBALANCE OF CELL PROLIFERATION AND APOPTOSIS IN ASPERGILLUS
    Ling-ling ZHOU; Zong-min WANG; Hui XIA; Zhi-guang ZHAO
    2009, 29(3):  292-295. 
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    Objective To observe the expression of cell proliferation and apoptosis in congenital pulmonary cyst and investigate their effect in the development of aspergillus Methods Immunohistochemical method was used to detect the expression of KI-67、BCL-2 and BAX in normal lung tissue from 10 adult (control group 1,CG1),relative normal lung tissue around the aspergillus from 20 aspergillus cases(control group 2,CG2)and abnormal lung tissue from 20 aspergillus cases. Results The expressions of KI-67,BAX and the ratio of BAX/BCL-2 in the bronchial epithelium of aspergillus were significantly higher than those in the two control groups(P<0.01)while the expression of BCL-2 was obviously lower(P<0.05).In aspergillus cases, the ratio of BAX/BCL-2 in IPA was much higher than that in Aspergilloma and ABPA (p<0.05). Conclusion The imbalance of cell proliferation and apoptosis in the bronchial epithelium of aspergillus played an important role in the development of aspergillus.
    Glucose effects on expression and function of 11β-hydroxysteroid dehydrogenase type 1 in pancreatic beta cell line NIT-1
    Mei LIN; Mu-xun ZHANG; Jian-hua ZHANG; Yi-kai YU
    2009, 29(3):  296-300. 
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    Objective To detect the glucose effect on expression and function of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in pancreatic beta cell line NIT-1. Methods The cellular localization of 11β-HSD1 was determined by immunocytochemisty in NIT-1 cell. The expression of 11β-HSD1 mRNA and protein were analyzed by RT-PCR and Western blot. The NIT-1 cell were cultured for seventy-two hours in Ham's F12 medium with 10mmol/L ,15mmol/L ,20mmol/L and 25mmol/L.The function of beta cell was evaluated by glucose-stimulated insulin secretion (GSIS) test. Results (1) The distributing of 11β-HSD1 was cytoplasm which was stained in brown. The expression of 11β-HSD1 mRNA and protein were obtained in NIT-1 cell. (2) With increasing of glucose in medium, the expression of 11β-HSD1 had higher levels and the secretion of insulin were decreased in NIT-1 cell. Conclusions The pancreatic beta cell line NIT-1 had expression of 11β-HSD1 gene. It suggested local glucocorticoid metabolism in pancreatic islet may be effect beta cell function.
    A low dose of Cyclophosphamide inreases the apoptosis of neutrophil at early postburn stage in rats
    Zai-rong WEI; Da-li WANG; Dan LUO; Yu-ming WANG; Bo;WANG; Guang-feng SUN; Jian-ping QI
    2009, 29(3):  301-304. 
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    ObjectiveTo study changes of the apoptosis of polymorphonuclear neutrophil at early postburn stage in rats and the interventional effects of low-does cyclophosphamide. Methods 110 SD-rats were divided into 3 groups randomly: experiment group(model+low dose of CY,n=50),burn group(model,n=50) and normal group(n=10). Drugs were injected into abdominal cavity immediately after burn , bone marrow and blood samples were obtained in the 3th,6th,12th,24th and 48th h.The apoptosis of PMN in the bone marrow and blood was analyzed by FCM. Results The apoptosis rate of PMN in the bone marrow: increased in burn group from 6th h (p<0.05), peaking at 12th h, cutted down from 24th h.; increased in CYgroup from 6th h (p<0.05),and continued to increase at 48th h.. CY group was compared with burn group , increased at 24th h. and 48th h. (p<0.05). The apoptosis rate of PMN in the blood : cutted down from 6th h (p<0.05),and to 48th h in burn group; cutted down from 6th h (p<0.05),and to 48th h in CY group; CY group was compared with burn group , increased at 6th h、12th h (p<0.05).Conclusion The low dose of CY could promote PMN apoptosis of the bone marrow and blood at early postburn stage. It could influence the development of burn SIRS.
    技术与方法
    Extraction and identification of postsynaptic density proteins from rat brain
    Ying ZHOU; Zhi-ling HUANG; Bo XIAO; Xiao-mei WU; Liu-yiwen WU; Pu YANG
    2009, 29(3):  305-308. 
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    Objective To purify postsynaptic density (PSD) proteins with better dissolubility. Methods The constituents of P2,synaptosome and PSD-1 were obtained by sucrose gradient centrifugation. The purity of PSD-1 protein was validated by Western blot. These soluble proteins PSD-2 were extracted from PSD-1 using the technology of membrane sequence extraction. The protein spots of PSD-1 or PSD-2 were seperated by two-dimensional gel electrophoresis(2-DE), and analyzed by the software of PDQuest in order to show the different dissolubility of PSD-1 or PSD-2. Results The results of Western blot showed that the postsynaptic marker PSD-95 were positive staining in P2, synaptosome and PSD-1, but the presynaptic marker synaptophysin was not seen in PSD-1. There were 286±25 protein spots in the electrophoregram of PSD-1, 720±17 protein spots in the electrophoregram of PSD-2, and the difference of these numbers was significant (P<0.01). Conclusions Pure PSD proteins can be obtained and dissolved easily by the method of sucrose gradient centrifugation plus membrane sequence extraction.
    Transfection of A549 human lung cancer cells can be mediated by nano-hydroxyapatite
    Qing-feng ZHENG; Jian-jun WANG; Min-gang YING; Shuo-yan LIU; Ying-chao HAN
    2009, 29(3):  309-313. 
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    Objective To evaluate whether the expressing vector pGenesil-1 encoding enhanced green fluorescent protein (EGFP) could be transfected into A549 human lung cancer cells mediated by nano-hydroxyapatite (nHAP) and to determine the transfection efficiency. Methods The nHAP was synthesized by the homogeneous precipitation method. The structure of the nanoparticles was observed under transmission electron microscope. The nHAP was prepared using ultrasonication and Na2CO3 and was modified with poly-L-lysine (PLL) at pH 7.4. The transfection of pGenesil-1 into A549 was divided into three groups as follows: nHAP-PLL group (n=8) mediated by nano-hydroxyapatite modified with poly-L-lysine (nHAP-PLL), liposome group (n=8) mediated by Lipofectamine and control group (n=8). After 48 hours transfection, the expression of EGFP was observed with fluorescent microscope. Flow cytometry was used to detect the transfection rate of each group. Results Under transmission electron microscope, the synthesized product presented needle-like and well dispersed particles with evenly distributed sizes of (15~20nm)×(60~80nm). Under fluorescent microscope, the expression of EGFP was detected in A549 cells of nHAP-PLL group and liposome group while no expression of EGFP was seen in A549 cells of control group at 48 hours post transfection. Flow cytometry analyses showed that, the transfection rate of A549 cells was (31.8±1.9)% of nHAP-PLL group and (33.4±3.7)% of liposome group. Compared with the control group, there was significant difference respectively (P<0.01), but there was no significant difference of transfection rate between nHAP-PLL group and liposome group (P>0.01). Conclusion The nHAP modified with poly-L-lysine can mediate the transfection of plasmid into A549 human lung cancer cells.
    研究短文
    EBP50 inhibits proliferation and matrix metalloproteinases expression of HeLa cells
    Jun-fang ZHENG; Kun LIU; Gui-ling MA; Peng CHEN; Jun-qi HE
    2009, 29(3):  314-315. 
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    Effects of aldosterone on the expression of endothelin subtype A receptor in rat cardiac fibroblasts
    Yu-zhou WU; Wei CUI; Shu-qin LI; Lei ZHANG; Jing-chao LU; Ji-dong ZHANG; Xiao-hong YANG; Jun DU
    2009, 29(3):  316-317. 
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    Heart-type fatty acid-binding protein down-regulation in rat's cardiocyte after acute myocardial ischemia
    Hong-yan WANG; Xiao-yun ZHAO; Han-ying XING; Ping ZHANG; Xia-hui LI
    2009, 29(3):  318-319. 
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    Expressions of Catenin-βand NF-κB in colorectal carcinoma and their clinical significance
    Xing DONG; Wei-dong ZHOU; Wan-jun YU; Wei ZHU
    2009, 29(3):  320-321. 
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    临床园地
    Significance of CD105 expression in laryngeal carcinoma
    Qing-ming LI; Bao-quan ZHANG; Pei-hong PENG
    2009, 29(3):  322-325. 
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    Objective To study the expression of CD105 in laryngeal carcinoma and evaluate the clinical significance of microvessel density(MVD). Methods Pathologic paraffin-embedded sections and clinic data from 40 patients with primary squmamous cell carcinoma of larynx were studied through immunohistochemistry. Microvessel density (MVD) highlighted by CD105 and CD34 were counted according to a standard protocol . Results CD105 expression in tumour tissue was significantly higher than normal health mucosa. The mean CD105-MVD value of T3 and T4 tumours showed a significantly higher staining than T1 and T2;The mean CD105-MVD value of tumours with metastasis was also markedly higher than tumours with no metastasis;The diffenerce of mean CD105-MVD value between in 16 patients with recurred laryngeal carcinoma and in 20 patients with unrecurred laryngeal carcinoma for two years of follow-up was markedly significant(P﹤0.05). Conclusion CD105 is a marker of tumour angiogenesis and an independent indicator of predicting invasion,metastasis and recurrence , and evaluating prognosis of malignant tumours.
    短篇综述
    Research Progress in Signal Pathway of Corneal Wound Healing in Diabetes Mellitus
    Ying ZHANG; Xin-yi WU
    2009, 29(3):  326-329. 
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    Delayed wound healing in diabetic cornea may be associated with the activation of MAPK signal pathway, impaired signal transduction of TGF-β signal pathway, abnormal expression of genes related to insulin signal pathway and reduced expression of insulin receptor. In addition, the activation of NF-κb and cytochrome C signal pathway also has harmful influence on delayed wound healing. This paper reviews research progress about signal pathway of delayed wound healing in diabetic cornea.
    Progress in molecular Biology of MELAS
    Yan-yan CAO; Yu QI
    2009, 29(3):  330-334. 
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    MELAS syndrome (mitochondrial myopathy encephalophathy with lactic acidosis and stroke-like episodes), as one of the most common diseases in mitochondrial encephalomyopathies, is characterized by highly variable manifestations. So, more and more people come to realize the importance of molecular basis of MELAS. This review took the commonest mtDNA point mutation (A3243G) for example to overview its molecular biological mechanism, test strategy and recent progress of study on MELAS syndrome.