1.Study on the Serum Levels of Placenta Growth Factor During Normal Pregnancy and in Patients with Pregnancy Induced Hypertension
Hong TENG ; Ping XU ; Ci WEI ; Bangrong JIANG
Journal of Jilin University(Medicine Edition) 2003;29(5):616-619
Objective:To detect serum levels of placenta growth factor(PIGF)throughout normal pregnancy and in patients with pregnancy-induced hypertension(PIH)as well as to explore the relationship between P1GF and pathogenesis of PIH. Methods: Serum specimens were collected from 51 healthy pregnant women as a control group and 33 women suffered from PIH as third trimester of normal pregnancy(P<0.001).There was a trend that serum levels of P1GF in PIH group decreased with the severity of PIH(P<0.001).Conclusion:PlGF play important roles in placental angiogenesis throughout pregnancy, and decreased serum levels of PlGF is associated with PIH.
2.Cloning of Mouse Enamel Matrix Serine Proteinase Encoding Mature Protein
Yabing MU ; Hongchen SUN ; Zebing ZHANG ; Jie OUYANG
Journal of Jilin University(Medicine Edition) 2003;29(6):705-709
Objective: To clone cDNA of enamel matrix serine proteinase (EMSP1) encoding mature protein from mouse dental germs. Methods: Total RNA was isolated from developing incisors and molars of 7 days mouse pups and reverse-transcribed into cDNA. Two pairs of specific primers was designed to obtain the desired gene by Touchdown PCR and Nested PCR. The segment was inserted into Vector pMD-18T, and recombined vectors was transformed into E.coli JM109.The positive clone was chose and analysed by restriction endonuclease mapping and DNA sequencing. Results:700 bp of cDNA of mouse EMSP1 was sueccessfully cloned from mouse tooth germs tissue. The sequence was consistent with that displayed in PubMed. Conclusion:The mouse EMSP1 cDNA encoding mature protein is obtained for further study.
3.Involvement of VLA-5 and VLA-6 in facilitating endothelium-oriented transmigration of hematopoietic stem/progenitor cells
Ling JIN ; Weizhong WANG ; Chunjiang LI
Journal of Jilin University(Medicine Edition) 2003;29(3):249-254
Objective:To investigate whether VLA-5 and VLA-6 are involved in facilitating en-dothelium-oriented transmigration of hematopoietic stem/progenitor cells. Methods:Purified hu-man CD34+ cells were subject to ex vivo transmigration assay and blocking experiments throughtranswell filter inserts coated with human umbilical vein endothelial cells (HUVECs). Four-colorfluorescence-activated cell sorting (FACS) analysis was applied to detect the expression profilesof adhesion molecules and chemokine receptor CXCR-4 on CD34bright cells. Results:Stromal cell-derived factor (SDF)-1-induced transmigrations of both mobilized peripheral blood (mPB)-(56.6±20. 1)% and bone marrow (BM)- (15. 6±1. 8)% derived CD34+ cells were significantlyfacilitated through HUVECs-coated transwell filter insters compared with noncoated ones, whichwere efficiently blocked by preincubation of CD34+ cells with neutralizing antibodies to VLA-5,or VLA-6, or both of them; meanwhile the proportions of migrating CD34+ cells through bothHUVECs-coated and noncoated transwell filter inserts in BM were significantly lower than thosein mPB; the different percentages of migrating CD34+ cells between in mPB and BM were corre-lated with their variable expressions of VLA-5 and VLA-6, but not for VLA-4 or chemokine re-ceptor CXCR-4. Conclusion:Facilitating HS/PCs transmigrations through HUVECs are involvedin both VLA-5 and VLA-6.
4.Evaluation of proliferative activities in Wilms'tumor
Lin WANG ; Xiaoyan ZHOU ; Yanan CUI ; Guizhen ZHANG ; Yuanyao CHEN
Journal of Jilin University(Medicine Edition) 2003;29(4):381-384
Objective: To assess the potential significance of proliferating cell nuclear antigen (PCNA) labelling indexes (PCNA-LI) and AgNORs number in evaluation of proliferative activity of Wilms′ tumor. Methods: A silver staining for AgNORs and an immunohistochemical method PCNA staining were performed on the biopsy specimens taken from 34 children with Wilms′ tumor. Results: No significant differences were observed between PCNA-LI and the pathological types and clinical stages, whereas fraction of S-phase and PI and the number of AgNORs were significantly higher in patients with PCNA-LI≥25% than that in patients with PCNA-LI<25%. The number of AgNORs/cell correlated well with both pathological types and clinical stages. The combination of PCNA-LI and AgNORs can accurately reflect the proliferative activity of cancer cells in Wilms′ tumor. Conclusion: The current pathological types and clinical stages may reflect the aggressive activity in Wilms′ tumor, but insufficient. The simultaneous determination of PCNA-LI and AgNORs count could be used as the essential complementarity of conventional pathological types and stages for accurate evaluation of biologic behaviour of Wilms′ tumor.
5.Application of EST Data of Siderophore Regulation Protein-like Gene of Aspergillus oryzae RIB40
Lin XIE ; Guizhi JU ; Goto KUNIYASU
Journal of Jilin University(Medicine Edition) 2003;29(1):1-4
Objective: To acquire the new clear color phenotype of Aspergillus oryzae b y th e antisense strategy of siderophore regulation protein (SREP)-like gene. Method s: Construct the cDNA library of Aspergillus oryzae RIB40 and amplify the fr agme nt ac7336f from the every EST clone, which had high homology with SREP gene of o ther species, then construct the eukaryotic expression vector with SREP-like ge ne using antisense strategy. Results: The sequence of this SREP-like gene was a cquired, the vector was successfully constructed. Conclusion: The deduced amino acid sequence of SREP-like gene of Aspergillus oryzae indicated that there is t he high homology with those of SREP genes of Penicillium chrysogenum, Neur ospora crassa and Schizosaccharomyces pombe.
6.The study on effect of long-termed administration of mixed rare earth Changle on rat liver
Ying LIU ; Dong CHEN ; Aijun CHEN ; Xiaoming WANG ; Yuxiu NIE
Journal of Jilin University(Medicine Edition) 2002;28(6):567-570
Objective: To research effect of different doses of Changle on the structure and function of rat liver.Methods: 180 healthy Wistar rats were divided into 5 experimental groups according following doses: 0.1,0.2,2.0,10.0 and 20.0 mg*kg-1,respectively, and the control group given physiological saline for six months. The changes of liver structure were examined by means of normal histological chemistry and transmission electron microscope(TEM). Results: The body weight of animal was linearly increased with the decrease of administered doses, gradual reduction of glycogen in hepatocytes and infiltration of inflammatory cells in the portal area were found in the group of 20.0 mg*kg-1. Changes of ultrastructure showed there were dense bodies and lysosomes containing dense granules in Kupffer cell and hepatocyte,and they were increased along with doses adding. Nuclei deformed, ALP and GPT in serum were rose in the group of 20.0 mg*kg-1. Different doses of Changle could lead to distinct biological effects. Conclusion: Long-termed administration of 20.0 mg*kg-1 Changle can lead to damage of structure and function of rat liver.
7.A novel peptides, like nerve growth factor, inducing pheochromocytoma PC12 cell differentiation
Xianhui LI ; Yawen LIU ; Guofa ZHAO ; Hui ZHU
Journal of Jilin University(Medicine Edition) 2002;28(3):221-225
Objective: To isolate the key domain of a novel polypeptide fragment from NGN-β that functions like intact NGF molecule.Methods: NGF-β had been treated with cyanogen bromide and trypsin skillfully. The peptide fragment with the activity inducing PC12 pheochromocytoma cells differentiation was isolated and purified by Sephadex G-50 gel filtration chromatography , DE-52 celluloseion exchange chromatography and C-18 reversed-phase column HPLC after NGF-β cleavaged by CNBr at 9th met then by trypsin at Arg or Lyscleavaged by CNBr at 9th met then by trypsin at Arg or Lys. Amino acid sequencing of this novel peptide fragment was performed by Automatic Amino Acid Analyser and Amino Acid Sequencer. Results: The functional fragment from cleavaged NGF might induce differentiation of PC12 cells. The fragment was consisting of two linear polypeptides . One of them was 16 peptide, GEFSVCDSVSVWVGDK , and other was 14 peptide, HWNSYCTTTHTFVK, linked by a disulphide bridge corresponding to residues 10~25 and 75~88, respectively, of the amino acid sequence of nerve growth factor, the result of biological activity assay in PC12 cells showed that the optimum concentration of this peptide were 0.001~0.1 μg*L-1. Conclusion: A novel peptide inducing differentiation of PC12 cell line of pheochromocytoma cells was obtained in the study. It′s isolation and purification successfully will underlie synthesis or expression of hyperactive neurotrophic small molecular substance although the relationship between the configuration and functions is not clearly.
8.Effects of whole body irradiation with X-rays on the expression of TfR on lymphocytes in spleens of mice
Can LUO ; Shibo FU ; Guizhi JU
Journal of Jilin University(Medicine Edition) 2002;28(1):1-5
Objective: To explore effect of TfR on immune function after ionizing by investigating changes in TfR expression on splenic lymphocytes of mice after whole body irradiation (WBI) with different dose X-rays. Methods: Direct immunoflurescence antibodies and flow cytometry were used to examine the changes of TfR expression. Results: The cell number of TfR positive expression in spleens increased significantly at 24 hours and 72 hours after WBI with 75 mGy X-rays but the cell number of TfR positive expression in spleens decreased significantly at 24 hours after WBI with 1~6 Gy X-rays. The activity of IL-2, meanwhile, demonstrated a parallel change. Conclusion: These results suggest that the TfR enhances immune function in low dose ionizing radiation but suppresses immune function in high dose. The change of TfR expression may be due to the change of IL-2 activity caused by ionizing radiation.
9.Expression of ATP synthase 6 in breast cancer cell line, T47D cells, induced by heregulin β1
Journal of Jilin University(Medicine Edition) 2002;28(2):111-114
Objective : To explore expression of related genes in breast cell line T47D induced by heregulin β1. Methods: Suppression subtractive hybridization was performed using cDNA sub traction kit to detect expression of heregulin-responsive genes in T47D cells. Results: ATP syn thase 6 was up-regulated by heregulin β1 in T47D cells at 1 and 6 hours. Conclusion: Heregulin β1 participates in the regulation of expression of both ATP synthase 6 and oxidative phosphoryla tion of T47D cells.
10.Study on relation among sICAM-1 level change in the patients with chronic viral hepatic disease,viral replication and the clinical significance
Dongfu LI ; Zhenjing JIN ; Xianming MENG ; Liulan PAN ; Liying WANG ; Jinghua TAI
Journal of Jilin University(Medicine Edition) 2001;27(4):374-376
Objective: To observe relation among sICAM-1 level in the sera of patients with chronic viral hepatic disease, viral replication and the clinical significance. Methods: The level changes of sICAM-1, IL-1,IL-8 and TNFa in the sera of patients with chronic hepatitis B(CH),chronic severe hepatitis B(CSH) and hepatocirrhosis (HC) were detected by ELISA. Results: The levels of sICAM-1,IL-1,IL-8 and TNFa in sera of the patients with chronic viral hepatic disease were significantly higher than those of healthy control group(P<0.01). The bilirubin level in the sera of patients with chronic viral hepatic disease of different clinical types positively correlated with sICAM-1 and cytokine activities. The sICAM-1 level in the sera of HBV-DNA or HBeAg positive patients was significantly higher than those of negative patients(P<0.05,P<0.01). Conclusion: The sICAM-1 level and activities of cytokine IL-1,IL-8 and TNFa as well as the bilirubin level in the sera of the patients can reflect the necrosis degree of hepatocytes, and the sICAM-1 level and activities of IL-1,IL-8 and TNFa were related to state of HBV carrier or the active degree of HBV in patients with hepatic disease.