Table of Content

25 June 2008, Volume 28 Issue 6

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研究论文

Screening and verification of the interactive proteins for nervous system polycomb 1

Yan-hua GONG; Lei SHI; Xu-dong WU; Bo-qin QIANG; Xiao-zhong PENG; Jian-gang YUAN

2008, 28(6):  529-534. 

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Objective To find out the protein complex members associated with the nervous system polycomb 1 (NSPc1) in vivo. Methods Use the yeast two hybrid system to screen the potential interactive proteins of NSPc1, and verify the interaction by GST pull-down, co-immunoprecipitation and subcellular co-localization assays. Results The full length, N-terminus and C-terminus of NSPc1 were used as the "baits" to screen the 3-month fetal brain cDNA library in yeast. Some proteins, including HBO1 (Histone Acetyltransferase Binding to ORC-1), were found to be interactive proteins of NSPc1, then the interaction between HBO1 and NSPc1 was verified both in vitro and in vivo. Conclusion The histone acetyltransferase HBO1 is one of the interactive proteins of NSPc1, and the interaction may be involved in the epigenetic repression activity of polycomb proteins.

Degradation of gelsolin in pancreatic cancer by ubiquitin-proteasome pathway

Xiao-guang NI; Xiao-hang ZHAO; Gui-qi WANG; Xiao-feng BAI; Fang LIU; Lan-ping ZHOU; Ping ZHAO

2008, 28(6):  535-538. 

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Objective: To explore the role of ubiquitin-proteasome pathway for the gelsolin protein degradation in pancreatic cancer. Methods: Pancreatic cancer cell lines BxPC-3 and PANC-1 were first treated with specific 26s proteasome inhibitor lactacystin. Immunoblots of cell lysates were probed for gelsolin expression. To determine whether gelsolin was conjugated to ubiquitin, proteins extracted from the cells treated with or without lactacystin were immunoprecipitated with anti-gelsolin antibody, followed by Western blot analysis using anti-ubiquitin monoclonal antibody. Results: The expression of gelsolin protein increased obviously after treated with lactacystin in BxPC-3 cells for 12h. Using anti-gelsolin antibody to immunoprecipitate gelsolin protein and followed by Western blot using anti-ubiquitin monoclonal antibody, it was found that inhibition of proteasome pathway by lactacystin resulted in accumulation of ubiquitylated forms of gelsolin protein. In PANC-1 cell line, there were no significant changes of gelsolin after treatment with lactacystin. Conclusion: Ubiquitin-proteasome dependent degradation may be an important regulatory mechanism for gelsolin down-regulation in pancreatic cancer.

Expression of β-NGF and p75NTR in vibrissa follicle bulge of developingr rat embryo

Sen LIN; Xiao-hua LIAN; Ting-ting LI; Min JIN; Yun WANG; Tian YANG

2008, 28(6):  539-542. 

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Object To detect the expression of β-NGF, p75NTR and hair follicle stem cell specific markers in vibrissa hair follicle of developing rat embryo. Methods Using frozen section and Immunohistochemistry to detect the expression of β-NGF and p75NTR, finally analysis data. Results The expressions of β-NGF and p75NTR in rat embryo vibrissa follicle change regularly. The expressions of three stem cell markers were different. Hair follicle stem cells (HFSCs) not only located in upper-hair follicle, but also in 3/4 of the lower part of the hair follicle in early stage of hair follicle morphogenesis. Conclusion The expression of β-NGF and p75NTR may be correlated with characteristics of hair follicle and hair follicle stem cells.

Expression and construction of an adeno-associated virus vector containing dual genes-GDNF and TH

Min ZHANG;

2008, 28(6):  543-548. 

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Objective Rat glial cell derived neurotrophic factor(GDNF) gene and tyrosine hydroxylase(TH) gene are widely used in gene therapy of Parkinson's Disease(PD).The present study is designed to investigate whether the two target genes GDNF and TH could be expressed simultaneously in one adeno-associated virus(AAV)vector and to discuss the probability of using it in gene therapy of PD. Methods RNA of GDNF and TH in HEK293 packaging cells were examined by reverse transcription-polymerase chain reaction (RT-PCR). Expressions of GDNF and TH in infected bone marrow stromal cells (BMSCs) and rat brain coronal sections were examined with immunocytochemistry and immunohistochemistry assay individually. Results RNA of GDNF and TH were detected in HEK293 packaging cells. Expressions of this two genes were observed in both bone marrow stromal cells and brain sections of rats. The expression efficiency of AAV-LacZ was about 50% in HEK293 packaging cells and approximately 15% in infected BMSCs and the expressions of TH and GDNF on the AAV-GDNF/TH virus injected points both had significant differences (p<0.01) compared with opposite injected with PBS. Conclusion Results of the present study suggested that GDNF and TH genes contained in one recombinant AAV vector could be expressed simultaneously both in vitro and in vivo and could provide references for further application in gene therapy of PD.

NAC on the improvement of insulin resistance induced by FFA in rats

Bing WANG; Hong-liang LI; Wen-ying YANG; Jian-zhong XIAO; Rui-qin DU; Xiu-ping BAI; Lin PAN

2008, 28(6):  549-552. 

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Objective:To investigate the changes of peripheral insulin resistance after lipid infusion and the effect of N-acetylcystein(NAC) intervention. Methods:Thirty-seven normal male SD rats, eight weeks old, were randomly divided into three groups, FFA group ,NS group and NAC group(using NAC 300mg·kg-1·d-1 two weeks before infusion). Catheters were implanted under pentobarbital anesthesia in the right atrium via the jugular vein and the left carotid artery. A technique for a 48-h infusion in unrestrained rats was used for triglyceride and heparin or saline infusion. The infusion period started on day 2 after surgery. After 48-h infusion, we determined free fat acid(FFA),nitrotyrosine,malonaldehyde(MDA),reduced glutathione hormone (GSH) level in plasma. The glucose infusion rat(GIR) was measured by hyperinsulinemia euglycemic clamp to evaluated the perpherial insulin resistance. The expressions of IRS-1,IRS-2 gene in muscle were detected by real time PCR. Results:(1)The FFA,nitrotyrosine and MDA concentrations in FFA group were higher than in NS group,but GSH level in plasma was lower.NAC intervention could reverse these effects.(2)The GIR was decreased significantly in FFA group compared with NS group (8.34±1.8mg.min-1.kg-1 vs 13.56±1.7 mg.min-1.kg-1,P<0.05), NAC intervention can reverse these effect(11.39±1.6mg.min-1.kg-1 vs8.34±1.8mg.min-1.kg-1,P<0.05).(3)The gene expression of IRS-1 was significantly decreased by 87.7% in FFA group, and the expressions of IRS-2 was decreased by 50.7% (all P<0.05).In contrast, the expressions of IRS-1,IRS-2 in NAC group reversed 370.1% and 46.2% respectively than in FFA group(all P<0.05). Conclusions:NAC intervension as a possible could increase the gene expression of insulin signal transduction molecules in muscle which might relate with the antioxidant effects of NAC.

Genetic polymorphisms of CYP1A1 and CYP2D6

Wen-kai LI; Qing-xia LIU; Fang-zhi CHEN; Ya-ping LO; Hui-juan CHEN; Jia-zhi XIA; Han-chun CHEN

2008, 28(6):  553-556. 

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Objective To study the correlations between genetic polymorphisms of cytochrome P450 CYP1A1 and CYP2D6 and leukemia predisposition by investigating the frequency distributions of genetic polymorphisms of CYP1A1 and CYP2D6 in leukemia patients and healthy controls in the population of Hunan Province in China. Methods The frequencies of polymorphic genotypes of CYP1A1 and CYP2D6 were detected by PCR and PCR-RFLP technologies. Results Frequencies of the wild (W/W), the heterozygous mutation (W/M) and the homozygous mutation (M/M) types of both CYP1A1 and CYP2D6 genes showed no significant differences between the patients with acute lymphoblastic leukemia (ALL), acute non-lymphoblastic leukemia (ANLL) or chronic myelogenous leukemia (CML) and the healthy controls. The individuals with W/M or M/M genotypes of CYP1A1 or CYP2D6 also showed no increased leukemia risk over their wild type counterparts. However, the frequency of CYP1A1W/M genotype combined with CYP2D6W/M genotype of ANLL group was higher than that of the control group. Conclusions The polymorphic variation of CYP1A1 alone or of CYP2D6 alone does not affect the susceptibility to leukemia. However, heterozygous mutation of CYP1A1 gene combined with that of CYP2D6 gene increases the risk of ANLL.

Screen plasmids targeted to Gcn5 and detect their effects on MSCs differentiation

Li Jing ZHU; Chuan FENG; Jie TIAN

2008, 28(6):  557-563. 

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0bjective To screen valid plasmid targeted to GCN5 among the constructed recombinant plasmids; and to explore the effects of histone acetylizad modification in regulating MSCs differentiation Methods Abstract the constructed plasmids and transfect them into MSCs induced by 5-aza. For 24h, observe MSCs under fluorescence microscope; detect the transfect efficiency by flow cytometry ; detect expression of protein GCN5 by Western-blot; detect the HAT activity by ELISA. Results Transfect efficiency was more than 20%; Expression of protein GCN5 and HAT activity had no difference in group ZJ1 and ZJ2, and had a significant difference in group ZJ3. The HAT activity of experiment group was significantly lower than that of all control groups. Conclusion The inhibitory state of histone acetylation results in plasmid ZJ3 , it can inhibit the differentiation process of MSCs. The results provide data for the clinical application of MSCs transplantation.

Repair of intestinal irradiation injury of Flk-1+ MSCs and SP cells

Qin HAN; Dong-nan HE; Yan-ning LIU; Kang-hua LI; Jie-shou LI; Chun-hua ZHAO

2008, 28(6):  563-569. 

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Objective: Comparing biological characteristics and the roles in repair of intestinal irradiation injury of bone marrow derived Flk-1+ MSCs and small intestinal derived SP cells. Methods: Phenotypes of the two cell population were determined by flow cytometery. Flk-1+ MSCs and intestinal SP cells were infused into irradiated mice respectively, and distribution and differentiation of these cells were determined. Histopathological examination and fibrosis degree was analyzed. Results: The phenotypes of Flk-1+ MSCs and intestinal SP cells were largely different. Both Flk-1+ MSCs and intestinal SP cells could home to injury rapidly, participate intestinal epithelial repair, and alleviate fibrosis and collagen deposition after irradiation. SP cells mostly engrafted into intestinal epithelium, while Flk-1+ MSCs partly engrafted into interstitial region as well as epithelium. Conclusion: Flk-1+ MSCs and intestinal SP cells could alleviate symptoms effectively, though their mechanisms are disparity.

Increased expression of phosphorylated α-synuclein in the substantia nigra of Parkinson's disease eat model

Lei-ming SUI; Huan-ying ZHAO; Chun-li ZHAO; Xiao-hong SUN;

2008, 28(6):  570-574. 

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Objective This experiment was to observe the effect of rotenone on phosphorylated α-synuclein and TH expression in the rat SN of PD. Methods Rats were stereotactically injected with rotenone. Movement distance and mean speed within 30-minute were calculated with Ethovison software. TH and phosphorylated α-SYN expression of the lesioned SN were demonstrated with immunohistochemical staining. The number of TH-positive neurons and phosphorylated α-SYN-positive neurons in the lesioned SN were measured. Results Movement distance and mean speed in the PD group were lower than those in the control group(P<0.05, 0.01). Compared to control group, TH-positive neurons counts in the lesioned SN in the PD group was reduced(P<0.01). The numbers of α-SYN-positive neurons were significantly higher in the PD group than in the control group(P<0.01). Conclusion Injection with rotenone could induce PD symptom, significant TH-positive neurons decrease and phosphorylated α-SYN increase.

Expressions of JAKs and SOCS1 in myocardium induced by acute exercise

Yun-hong WANG; Yang WANG; Li-li XI; Wen-jin WANG; Li-qin ZHANG; Chao-shu TANG

2008, 28(6):  575-578. 

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Objective To evaluate Janus kinase(JAK)s and SOCS1 expression in myocardium induced by acute exercise in order to understand the possible regulating mechanism of SOCS1 in JAK/STAT signaling pathway in exercised heart. Methods Male Sprague-Dawley rats were submitted to continuous graded treadmill running, then were sacrificed at 0, 1, 3, 5, 12 and 24 h respectively after the exercise. JAK1, JAK3 and SOCS1 expressions in the myocardium were determined with immunohistochemistry. Results JAK1 expression significantly increased immediately after running in comparison with control group, peaked at 1~5 h, and remained high at 24 hr after exercise; in contrast, JAK3 and SOCS1 expressions significant decreased from 1h to 5h after running, then gradually back to sedentary level 12h after exercise; Conclusion Acute exercise induced the different responses of JAK1 and JAK3, which indicated the different function of JAKs in exercised heart, SOCS1 may involve in regulating JAK/STAT3 signal pathway in the exercised heart.

Attenuated endocytosis function by down-regulation of cortactin protein in cancer cells

Li CHEN; Jian-wei ZHU; Xi ZHAN

2008, 28(6):  579-582. 

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Objective To study cortactin function in cancer cell endocytosis. Methods We applied cortactin siRNA interference to MDA-Mb-231, a human breast adenocarcinoma cell line, in which cortactin is over-expressed, and introduced anti-cortactin immunoreagents into the cells using the BioPorter system to interfere with cortactin function in vivo. Capture -ELISA assay was used to measure transferrin uptake, immunoblot assay to assess the effect of cortactin knock-down, and immunoflurescence microscopy to examine the effect of cortactin down-regulation on transferrin uptake. Results Interference of cortactin function in cells led to the impairment of transferrin endocytosis. Transferrin fluorescent intensity in cytoplasm in cortactin siRNA treated-cells was apparently reduced in comparison to mock-treated cells. Less than 50% of cells subjected to cortactin siRNA treatment had normal transferrin uptake. Endocytosis in MDA-Mb-231 cells introduced with cortactin antibodies was impaired as well, showing a 30 ~ 60% reduction in transferrin uptake. Conclusions This study implies that cortactin, an actin-binding protein, plays an essential role in cell endocytosis

L-3-n-butylphthalide inhibits the protein expression of nuclear factor-κB and IκB-α in primary cultured mouse brain glial cells following oxygen-glucose deprivation/ reoxygenation

Shan-shan DENG; Yong LO

2008, 28(6):  583-587. 

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Objective：To observe the effect of l-3-n-butylphthalide（l-NBP）on the protein expression of nuclear factor-κB and IκB-α in primary cultured mouse brain glial cells following oxygen-glucose deprivation/ reoxygenation（OGD/R）. Methods：Cultured mouse brain glial cells in vitro and established OGD/R model. Glial cells were divided into five groups: normal control（NC）group, OGD24h/R 24h group，l-NBP 20μmol/L group，l-NBP 100μmol/L group，l-NBP 500μmol/L group. The protein expression of NF-κB in nuclear and IκB-α in endochylema were analyzed by Western blot. Results：Comparing with NC group, OGD24h/R 24h group had a significantly higher expression of NF-κB (P<0.01）. The expression of NF-κB in l-NBP 100μmol/L group and l-NBP 500μmol/L group were evidently lower than that in OGD24h/R 24h group（P<0.01）. The degradation of IκB-α in OGD24h/R 24h group was significantly higher than that in NC group（P<0.01）. It was evident that the degradation of IκB-α in l-NBP 500μmol/L group was lower than that in OGD24h/R 24h group（P<0.05）. Conclusions：l-NBP can inhibit secondary inflammation of glial cells following OGD/R by decreasing the degradation of IκB-α and inhibitting activation of NF-κB.

Expression of Recombinant Human Endothelial Nitric Oxide Synthase gene in Lung tissue of mouse

Zhu-xiang ZHAO; Bing LI; Pi-xin RAN

2008, 28(6):  588-594. 

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Objective：To detect whether HENOS we cloned could be expressed in mouse lung after intratracheal administration of recombinant adenovirus vector. Methods：HENOS cDNA were obtained by RT-PCR from total RNA which extracted from human HUVEC. The replication-deficient heNOS recombinant adenovirus vector was constructed with a ligation method in vitro. The high titer of recombinant adenovirus was obtained by chromatographic methods. The expression of HENOS protein was determined by immunohistochemistry staining after intratracheal administration of recombinant adenovirus. Results：Sequence confirmed the cloned cDNA containing the whole ORF and the HENOS cDNA was about 3731bp in size.It showed 99.93% identity with that of HENOS cDNA in GenBank（*163729）.The biological activity of HENOS protein was examined in transfected cos7 cell line with HENOS cDNA in eukaryotic expression vector of pcDNA3.0. The replication-deficient recombinant adenovirus vector containing HENOS cDNA was constructed successfully and the purified viral titer was 2.0×1010pfu/ml. After intratracheal administration of the recombinant adenovirus, HENOS expression was detected in the majority of bronchial epithelium, alveolar lining cells, endothelial cells and smooth muscle cells of pulmonary vessels. In control, little endogenous eNOS immunoreactivity was detected in pulmonary vessels and it was no eNOS immunoreactivity in bronchial and alveolar epithelial cells. Conclusion：The replication-deficient recombinant human endothelial nitric oxide synthase mediated by adenovirus vector constructed could be delivered into the lung tissue of mouse by intratracheal administration of recombinant adenovirus and can be expressed in lung tissue in high-efficiency.

Pathological and immunological changes on SARS-CoV infected human ACE2 transgenic mice

Xiu-hong YANG; Yun-xin CHEN; Hua ZHU; Lan LAN; Chun-mei MA; Ya-li LIU; Chuan QIN

2008, 28(6):  595-599. 

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Objective To establish susceptible animal model of SARS, we studied the pathological and immunological changes on SARS-CoV infected human ACE2 mice. Methods After copy number of human ACE2 transgenic mice was determined by Real-time PCR, the animals were infected with PUMC01 strain of SARS-CoV through nasal cavity. The systemic pathological changes were observed under microscope. The specific antibodies in serum and cytokines of TNF-α, IL-6 and IFN-γ in the supernatant of lung homogenates were detected by ELISA. Results The human ACE2 transgenic mice with a single copy showed more severe interstitial pneumonia accompanied by many extra-pulmonary organ damages. The specific antibody was found in a few of transgenic mice. Levels of TNF-α, IL-6 and IFN-γ in the supernatant of lung homogenates from transgenic mice were increased more markedly after they were inoculated with SARS-CoV. Conclusion Resembling human SARS case, the human ACE2 transgenic mice with infected by SARS-CoV had similar pathological and immunological changes. This small animal model can be used to study pathogenesis of SARS and evaluate the effect of anti-SARS-CoV drugs.

Effect of propofol and midazolam on memory during target control infusion for

Liang ZOU; Xiang QUAN; Shou-yuan TIAN; Tie-hu YE

2008, 28(6):  600-603. 

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Objective To study the effect of propofol and midazolam on memory during target control infusion for sedation. Methods Forty healthy male volunteers were randomly devided into 4 groups, all subjects received target controlled infusion with propofol or midazolam for sedation.Group 1 and group 3 were sedated to OAA/S 1 or OAA/S 3, respectively,by using propofol ; group 2 and group 4 was sedated to OAA/S 1 or OAA/S 3, respectively by using midazolam. All subjects learned a list of words before sedation and at the different sedation depths predicted. After recovery the explicit and implicit memory were tested using word stem completion method and process dissociation procedure . Results Compared with 0, there was no significant difference in explicit memory among all the 4 groups. While there was significant difference in implicit memory for group 3 and 4,and there was no significant difference in implicit memory for group1 and group2. Conclusion Sedative-hypnotic drugs not only deepen sedation,but also impair memory as serum concentration increases during target infusion .Drug-induced amnesia is independent of sedation .Propofol and midazolam have the same efficacy on memory at the same sedation scale.

Differentiation of fetal-bone mesenchymal stem cells induced by three kinds of connective tissue in vitro

Yan YANG; Wei-wei WANG; Rong JIANG; Zheng DUAN

2008, 28(6):  604-609. 

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Objective To compare the inducing effect of three kinds of connective tissue from amnion, fetal mesentery and intestinal submucosa on the differentiation of fetal BMSCs in vitro. Methods The epithelium on three kinds of connective tissue were removed. and EGF and IGF-1 in these tissue clumpes were detected .The DAPI-P3-BMSCs were planted on the surface of three kinds of connective tissue to culture. There were connective tissue induced groups, connective tissue and EGF and IGF-1 induced groups, EGF and IGF-1 induced as control group. The tissue clump was collected on cultured 8d、10d、12d, to compare their inducing effect on the differentiation of BMSCs. Results All of BMSCs induced by three kinds of connective tissue and growth factors could express CK or EMA. The effect of unite-induced groups was stronger than those groups induced only by connective tissue, P＜0.05; The strongest inductive effect of three connective tissue was exerted by amnion, P＜0.05. EGF was expressed in the three kinds of connective tissue, and IGF-1 was only expressed in mesentery and intestinal submucosa. Conclusion BMSCs could be induced to differentiate into epithelial cells by the connective tissue of amnion, mesentery and intestinal submucosa, EGF and IGF-1 in vitro; EGF, IGF-1 could increase the effects of these connective tissue on the induction of BMSCs differentiation; EGF and IGF-1 may be one of the endogenic factor in these connective tissue which induces BMSCs to differentiate into epithelium.

Zinc finger protein 216 up-expressed in hepatocellular carcinoma tissue and sera

Xiang-fang GU; De-cai YU

2008, 28(6):  610-613. 

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Objective To find antigens related with hepatocellular carcinoma (HCC). Methods Human Zinc Finger Protein 216 (ZNF216) was identified by SEREX(serological analysis of recombinant cDNA expression libraries, SEREX). The 6*His fusion protein of ZNF216 was obtained by synthesis bacterially and purified with a nickel affinity chromatography column. The level of anti-ZNF216 antibody in the sera was detected by ELISA，and ZNF216 mRNA level in HCC tissue and its peripheral normal tissue was detected by RT-PCR. Results The level of anti-ZNF216 antibody in the sera of HCC patients was higher than healthy individuals. ZNF216 mRNA level in HCC tissue is evidently higher than peripheral normal tissue. Conclusion The over-expression of ZNF216 in the HCC sera and tissue indicates that ZNF216 perhaps plays a role in HCC, but still needing more evidence.

ShRNA inhibited SURVIVIN expression in human lung cancer cell A549

Run-xiu WANG; Fu-hua XIE; Xian-hu THANG; Bin WU; Nian-ci LIANG

2008, 28(6):  614-617. 

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To construct expression vector with short hairpin RNA of survivin，which paves the way for lung cancer RNAi-mediated therapy. Methods Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized.After annealed,the double Objective To construct shRNA expression vector of SURVIVIN for RNAi-mediated therapy on lung cancer. Methods DNA templates of SURVIVIN shRNA were designed,synthesized and cloned into the shuttle vector to get recombinant plasmids. After plasmids were transfected into A549 cells, the one with the most repression was screened by means of RT-PCR. Then MTT and Western blot were done to ascertain whether the proliferation of A549 cells were inhibited and SURVIVIN was down-regulated.Results The recombinants were constructed and screened successfully. The proliferation of A549 cells and SURVIVIN expression were repressed. Conclusion Mediated by RNAi, SURVIVIN expression was down-regulated,which theoretically based on gene therapy on huaman lung cancer.

技术与方法

Establishment of an auditory deprived mouse model

Yong-sheng TIAN; Meng ZHANG; Xiao-wei CHEN; Xing AI

2008, 28(6):  618-622. 

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Objective To explore a method for establishing a stable auditory deprived animal model of mouse. Methods Forty two-month old BALB/c mice with normal auditory brainstem response (ABR) thresholds were divided into two groups randomly. The experimental group mice were underwent an incision inferior to the pinna, and the cochleas were destoried with a drill through the bulla bilaterally. The control group mice were underwent only an incision without cochlea ablation. Mice were tested for ABR thresholds 4 month later to assess hearing sensitivity by comparing the ABR before and after operation. Results In the experimental group, mice's auricle reflection disappeared after operation and ABR waves couldn't be detect. In the control group, the tests of mice's hearing were normal. Conclusions Mouse cochlea ablation is a reliable method for establishing a stable auditory deprived animal model of mouse by the pinna inferior approach. It can be used to research auditory centre plasticity induced by auditory deprived.

Preparation and identification of monoclonal antibody against human SUMO1

Lin WANG; Feng-wei TAN; Shi-ping CHEN; Li-fang LU; Yan-hua GONG; Xiao-zhong PENG

2008, 28(6):  623-626. 

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Objective To express the human recombinant SUMO1 protein and prepare monoclonal antibody (mAb) against it. Methods The recombinant expression plasmid pET32a-HIS-SUMO1 was made and transformed into E.coli (BL21), and then the recombinant fusion protein HIS-SUMO1 was expressed and purified. The BALB/c mice were immuned with pure protein HIS-SUMO1 as antigen. Monoclonal antibody against SUMO1 were prepared by using standard hybridoma technology. The hybridoma cell lines were obtained by ELISA and Western Blot screening procedure, the isotype of the mAbs were further identified by immune-double diffusion. Ascites were prepared from one propagated hybridoma cell line and mAbs were purified by using the Kit of Millipore. The valence of mAb was detected by Western Blot. Results The recombinant protein HIS-SUMO1 is expressed and purified . Three hybfidmas producing antibodies against SUMO1 were obtained, the isotypes of three mAbs are IgG1, Western Blot showed that the antibodies were specific for SUMO1. The antibody purified from the ascites has better specificity. Conclusion The SUMO1 mAb prepared by using recombinant SUMO1 protein as antigen can be used for detecting the protein sumoylation.

研究短文

Bee venom improving rhythmic contraction of isolated toad heart

Ying JIN; Zhi-jie LIN; Hui-di YANG; Yan-ping LAI; Qing JI; Yan-qiang LIU

2008, 28(6):  627-628. 

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短篇综述

Investigation progress of proprotein convertase subtilisin/kexin type 9 influencing the metabolism of cholesterol

Wen-long LI; Wei-dong PEI

2008, 28(6):  629-632. 

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Familial hypercholesterolemia (FH) is a common autosomal dominant dyslipidemia, which caused by mutations of the low-density lipoprotein receptor (LDLR) gene producing defect or deficiency in LDLR. The characters of FH are elevated level of total and LDL cholesterol. FH is considered to be a complex polygenic disease. Recently, more findings indicate that proprotein convertase subtilisin/kexin type 9 (PCSK9) gene play an important role in serum cholesterol metabolism. Some mutated PCSK9 proteins decrease LDLR, which cause FH in affected families. Some other mutated PCSK9 proteins decrease self-affinity, which cause hypocholesterolemia. We review the newest researches about the structure, function of PCSK9 gene and the relation of its mutations with plasma cholesterol metabolism.

Mechanisms studies on myocardium repairing by stem cell therapy

Ying WANG; Jin ZHAO; Xiao-cheng LIU

2008, 28(6):  633-637. 

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Stem cells transplantation has shown bright prospect in clinic myocardial infarction therapy. The mechanisms of it have experienced several stages, such as: cell differentiation, angiogenesis et al, which offered us a new clue for therapy. This review focuses on mechanisms of stem cells transplantation.

Research update on bleomycin-induced systemic sclerosis

Kun XIAO; Xuan ZHANG

2008, 28(6):  638-641. 

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Systemic Sclerosis(SSc) is a fibrotic disease characterized by immunological abnormalities，vascular injury and increased accumulation of extracellular matrix. The etiology has not been fully elucidated yet, so we need to establish a suitable animal model for research. This review is to introduce murine models of bleomycin-induced SSc, and summarize research works on the pathogenesis and treatment of SSc based on this model.

PIG7 gene and human diseases

Jia-zhuo LIU; Jian-xiang WANG

2008, 28(6):  642-645. 

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Apoptosis induced by P53 gene is one of the most important mechanisms of signal integration in cells，which identify the cell injury and start downstream apoptotic signals. PIG7 gene is induced by P53 and correlates with metabolism, inflammation and tumor. In this review, we mostly described the recent findings in the molecular biology of PIG7 gene and the correlation with human diseases.

临床园地

Diagnosis and treatment of stationary paraganglioma in retroperitoneum

2008, 28(6):  646-650. 

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Object To highlight the diagnose and treatment of stationary paraganglioma in retroperitoneum . Methods 10 cases of stationary medullary chromaffinoma of retroperitoneum proven pathologically from 1997 to 2007 were reviewed and sutdied . Hypertension were not observed in all patients , who were admitted to hospital through routine phisical examination . Endocrine secretion examinations , B-US , CT, MRI , 131ＩMIBG ,and octreotide were used to dianose the diease.Among all 10 patients , 4 cases took α-receptor blocker for 2-4 weeks preoperatively.3 were operated laparoscopically , 7 were operated with lumber incision .Results Hypertension and urinary catecholamine was usually normal or elevated lightly in stationary medullary chromaffinoma of retroperitoneum .131and octreotide have high sensitivity and accuracy in diagosing medullary chromaffinoma . 4 were near renal hilum , 6 was in para-aorta.　They were from 3cm to 15cm in size . The results of pathology were paraganglioma in 7 and malignant paraganglioma in 3 . Tumor markers were positive ，such as cgA ，syn， NSE and s-100.There was 1 case metastasis 1 years later .Conclusions Once the correct diagnosis is acquired , surgical treatment should be carry out on the basis of correct drug preparation . Intimate follow-up is necessary and important .

医学教育

Usage and prospect of digital-internet microscopic mutual motivation system for the experimental morphology teaching

Hong-xing SU; Hong-wei SHANG; Li-xin ZHANG; Xin LU; Peng JING; Juan DU; Xiao-li DONG

2008, 28(6):  651-653. 

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Accompanying with highly developed computer and internet technology in the 21st centry, there has been a revolutionary change in the medical experimental morphology teaching. For matching to this change, Capital University of Medical Science has appropriated more money on experimental teaching, and established fully-digital multi-media internet mutual motivating classroom. It has been changed that teaching procedure were carried out by just anatomy model and microscope, which could change the teacher`s role and teaching formality dramatically. As a result of this extraordinary change, it not only made the teaching effect become more efficient, but also made the students` ability of cognizance, problem analysis and general competency better.

Optimizing Chinese medical rducation evaluation system through comparing experiences from overseas

Bai-hong GAO; Wei-wei XU

2008, 28(6):  654-656. 

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It's very important to establish an evaluation system to match international standards for improving the medical education quality in China. This article reviews the global profile of evaluation process for higher education, and compares it with the current status in China. It introduces the characteristics and experiences from overseas as references to optimize our medical education evaluation system.