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.

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.

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.

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.

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.

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.

Objective:To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism.Methods:The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells;the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method;the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits.Results:The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation,the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation,the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation.Conclusion:ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

Objective:To explore the effects of voriconazole on the proliferation and morphology of Acanthamoeba cultivated in vitro,and to clarify the killing effects of voriconazole against the trophozoites and cysts of Acanthamoeba.Methods:The Acanthamoeba polyphaga at logarithmic phase were selected and divided into control group and experiment groups(2.5 and 25.0 mg·L-1).The Acanthamoeba in each group was collected at 24,48,72,and 96 h after drug administration,respectively.Then the concentrations of Acanthamoeba were calculated and the proliferation curves were drawn;inverted microscope was used to observe the morphology,activity and adherence of Acanthamoeba;the ultrastructures of Acanthamoeba were observed under electron microscope.Results:Compared with control group,the numbers of Acanthamoeba polyphaga in experiment groups were significantly decreased(P<0.01).The morphology of Acanthamoeba changed significantly under inverted microscope,and the shape of Acanthamoeba transformed from the trophozoites with irregular spiny filopodia to circular cysts.Even a large number of cell debris was observed.Different degrees of damage and even necrosis of Acanthamoeba in experiment groups were found under electron micro scope.Conclusion:Certain concentration of voriconazole can effectively inhibit the proliferation of Acanthamoeba and change the morphology and ultrastructure and kill the trophozoites and cysts of Acanthamoeba cultivated in vitro.

Objective:To investigate the effect of the extract of Schisandra chinensis on the matrix metalloproteinases(MMPs) system in kidney tissue of the diabetic rats,and to explore its protective effect on the kidney tissue from the matrix degradation perspective.Methods:STZ was used to establish rat models of diabetes mellitus.A total of 45 diabetic rats were randomly divided into model group,extract of Schisandra chinensis group and Benazepril group,and there were 15 rats in each group.Another 15 rats were selected and used as normal control group.12 weeks after administration,the routine blood and urine biochemical indexes,the histological changes,blood glucose (BG),blood urea nitrogen(BUN),serum creatinine(Scr),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterin(LDL-C),total cholesterol(T-CHO),and triglyceride(TG) levels,excretion rates of albuminuria and proteinuria of the rats in various groups were detected;the expression amounts of fibronectin (FN),type Ⅳ collagen (Col Ⅳ),and matrix metalloproteinase inhibitor metalloproteinase-2 (TIMP-2) in kidney tissue of the rats were detected by immunohistochemical method;the activity of matrix metalloproteinase-2 (MMP-2) was detected by zymography.Results:Compared with model group,the glomeruli matrix accumulation of the rats in extract of Schisandra chinensis group was significantly improved,the excretion rate of albuminuria,LDL-C level and serum MDA level were decreased(P<0.05),the activities of CAT(P<0.01)and SOD(P<0.05)in kidney tissue were increased,and the level of MDA in kidney tissue was decreased(P<0.05).The immunohistochemistry results showed that compared with model group,the expression amounts of FN,Col Ⅳ,and TIMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group were significantly decreased.The zymography results showed that compared with model group,the activity of MMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group was significantly increased(P<0.05).Conclusion:Extract of Schisandra chinensis has protective effect on the kidney tissue of the diabetic rats induced by STZ,and the mechanism may be related to the inhibition of oxidative stress and the improvement of MMP-2 activity as well as the inhibition of TIMP-2 expression which could improve the matrix degradation.

Objective To investigate the construction of compound membrane with corneal stromal cells and collagen-chitosan by tissue engineering technique and its biocompatibility.Methods Rabbit and human corneal stromal cells were separated and seeded into collagen-chitosan membrane.The compound membrane was transplanted into rabbit corneal stroma.Then the growth condition of keratocytes,the effect on normal keratocytes and degradation of compound membrane were detected by corneal confocal microscope,anterior OCT and histological and immunohistochemical methods ex vivo 1,2,4 weeks after grafting.Results The rabbit and human corneal stromal cells grown well in collagen-chitosan scaffold.The compound membrane degradated gradually after grafting.There was no necrosis and dissolvation.Corneal epithelium,stroma and endothelial cells were all normal.Conclusion Collagen-chitosan can be used as a biological scaffold for construction of corneal stroma.Corneal confocal microscopy and anterior OCT are new methods to observe the biological activity of constructed corneal stroma.