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In order to study the influence of different gelatin concentrations, and lymphocyte isolation liquid on primary culture of umbilical cord blood-derived adhesive cells (hCBACs), the red blood cells of umbilical cord blood was separated by 3% and 6 % gelatin for detecting the effectiveness of sedimentation, then the adhesion rate at 48 hours, the day of initial expansion and the rate of culture success were detected for hCBACs cultured with CD34(+) cells after the mononuclear cells were separated by 6% gelatin followed by Ficoll and Percoll, and the morphological characteristics and growth status were observed by invert microscopy. Cytochemistry stain for nonspecific esterase stain (NSE), peroxidase (POX), periodic acid Schiff reaction (PAS) and alkali phosphatase (ALP) and immunocytochemistry labeling for CD31, CD45, CD68 and fibronectin (Fn) were detected. The results showed that 6 % gelatin was better than that 3% gelatin for red blood sedimentation. The Percoll was predominant over Ficoll in adhesion rate at 48 hours, the day of initial expansion, the time of initial formation of adhesive cell colony units, the time of maximal numbers of adhesive cell colony units, the the cell fusion time and ratio of culture success. 60% fibroblast-liked cells, 36% macrophage liked cells and 4% small-round cells were observed in cells isolated by both isolated methods. The cytochemistry stain for NSE, POX, PAS and ALP was similar in two groups, the difference was not statistically significant between these two groups. The immunocytochemistry labeling for CD31, CD45, CD68 and Fn was also similar in both groups and the difference was also not statistically significant between these two groups. It is concluded that the combination of 6% gelatin with Percoll is an ideal separation method for primary culture of hCBACs, which provides basic information for clinical application.

The microbial number, microbial biomass, and enzymatic activities in five upland soils under agricultural utilization for 50-700 years were determined, with the correlations between soil microbiological characteristics and agricultural utilization duration analyzed. In the meantime, the functional diversity of microbes in soils having been utilized for 50, 100, and 700 years were investigated. The results showed that at the early stage (< 100 years) of agricultural utilization, the number of soil fungi (F) had a slight increase, while the bacterial number (B), B/F ratio, microbial biomass C (C(mic)), microbial biomass N (N(mic)), and the activities of catalase, invertase and urease all decreased markedly. After utilized for more than 100 years, the F decreased significantly, while the B, B/F ratio, C(mic), N(mic), and the activities of test enzymes all tended to increase. During the whole utilization period from 50 to 700 years, the C(mic)/N(mic) ratio had a significant increase with year. The Shannon, Simpson, and McIntosh indices of soil microbial community had the same responses to the agricultural utilization duration as the bacterial number, microbial biomass, and enzymatic activities. All of these indicated that in the upland fields in Cixi of Zhejiang Province, shifts of soil microbial community occurred with increasing agricultural utilization duration, and soil microbiological quality had an overall increase after 100 years agricultural utilization.

This study was aimed to investigate the effect of vcam-1 gene-modified human umbilical cord blood derived stromal cells (CBDSCs) on hematopoietic regulation so as to establish the experimental foundation for further study. The target gene vcam-1 was cloned into the shuttle plasmid with the report gene GFP. The recombinant shuttle plasmid was transformed into BJ5183 bacteria to recombine with backbone vector pAdeasy-l, and the recombinant adenoviral vector ad-vcam-1-gfp was confirmed after transfection with CBDSCs. The results indicated that two fragments of about 9 kb and 2 kb were obtained after digestion of recombinant plasmid pAdTrack-vcam-1 with NotIand XhoI, and single fragment of 600 bp was obtained after amplification with PCR; two fragments of about 31 kb and 4 kb were obtained after digestion of recombinant plasmid pad-vcam-1-gfp with PacI, which suggested a successful homologous recombination. The expression of vcam-1 gene in ad-vcam-1-gfp transfected CBDSCs could be detected by immunocytochemistry, RT-PCR and fluorescent microscopy. It is concluded that the recombinant adenoviral vector ad-vcam-1-gfp has been constructed successfully, and the expression of vcam-1 is up-regulated in CBDSCs transfected by gene ad-vcam-1-gfp.

This study was aimed to investigate the expression of SDF-1 mRNA in SDF-1 cDNA-modified human bone marrow mesenchymal stem cells (hBMSCs) before and after transfection. The hBMSCs were isolated, cultured and identified, the SDF-1-pIRES2-EGFP eukaryotic expressing vector was constructed, and then the hBMSCs were transfected with the vector encapsulated by lipofectamine 2000. The transfection efficiency was measured by observing the expression of green fluorescence protein and detecting the mRNA by RT-PCR. The results indicated that the expression of SDF-1 mRNA increased by about 20% after hBMSCs were transfected instantaneously by SDF-1-pIRES2-EGFP. It is concluded that SDF-1 cDNA eukaryotic expression vector can be instantly transfected into hBMSCs by lipofectamine 2000, but the efficiency was too low to obtain enough steady transferred hBMSCs. Other procedures should be trialed to improve the transfection efficiency.

A pot experiment was conducted to examine the roles of earthworm in As uptake from original As-polluted soil by maize (Zea mays L.), and their effects on As, P fractions in the rhizosphere. The As-polluted soils with three As levels were collected from the arable soil near As mine. The plants were harvested after 10 weeks of growth. Dry weight (DW) and P, As concentrations of plants, as well as As and P fractions of the rhizospheric soil, were determined. The results showed that inoculated earthworm or appended rice straw increased maximal 149%, 222% DW of root and shoot, respectively. At the medium and high soil As levels, root As concentration in the soil treated by earthworm and rice straw was highest among all treatments, and earthworm increased more As concentration of shoot than rice straw did. In different soil As levels, root P concentration in the soil treated by earthworm was highest, and shoot P by rice straw. Ca-P affected maize absorbing As at the low soil As level(r = 0.981), and maize absorbing Al-P was restrained by As involved in well-crystallized hydrous oxides of Fe and Al at the medium (r = 0.953)and high (r = 0.997)soil As levels. The concentration of non-specially absorbed As and As combined with Fe or Al and of O-P increased at the soil inoculated earthworm or/and appended rice straw at the same time. These results indicated that earthworm was more valuable for plant developing than rice straw was.

Objective: To construct cell adhesion mediated drug resistance (CAM-DR) model based on Acute lymphocyte leukemia bone marrow stromal cells(BMSC) for further studying drug resistance of leukemia., Methods: Firstly, we adhesively cultured Jurkat cell strain of human leukaemia lymphocyte with the matrix cell radiated by (60)Co to construct the model of CAM-DR, then evaluated the model in morph and construction by scanning electron microscope. The IC50 of DNR on Jurkat cell was examen by MTT and the concentration and distribution of DNR in the cell was detected by flow cytometry., Results: When the Jurkat cells were cultured with BMSC for 24 h, we found that Jurkat cells had adhesioned the bone marrow stromal cell layer by parapodium and some of them had nichd the mesh consisted by the confluence of BMSC. At the point of 48 h, some Jurkat cells had migrated to underlayer of BMSC, and Jurkat cells were nichd the mesh of BMSC like nidi. The accumulation and distribution of DNR in the Jurkat cells were not affected in the model, but the reaction of Jurkat cells to DNR were significantly inhibited., Conclusion: The model of CAM-DR based on Acute lymphocyte leukemia bone marrow stromal cells was successfully built.

The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.

The aim of this study was to evaluate the effect of dimethyl amiloride (DMA), a specific inhibitor of Na(+)/H(+) exchanger-1 (NHE-1), on intracellular pH value (pHi), proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses, pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses, pHi decreased, the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile, after treatment with DMA during 100 micromol/L to 150 micromol/L, the increase amplitude of G(0)/G(1) phase cells and the decrease amplitude of S + G(2)/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification, the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells, and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-60 cells.

Soil samples of current paddy soils, ancient paddy soil/ancient dry land soil, and bottom soils were collected from two sites in Chuodunshan Site of Majiabang Culture (about 6,000a). 15 polycyclic aromatic hydrocarbons (PAHs) were analyzed with HPLC, and their possible sources were identified. The sum of 15 PAHs was 202.9microg x kg(-1) and 207.7microg x kg(-1) in the surface soils from Site A and Site B respectively, which were mainly deposited from atmosphere. In ancient paddy soil from Site A, the total PAHs concentration sharply decreased to 56.0microg x kg(-1), but was still higher than those in ancient dry land soil and bottom soils with the sum of 32.0 approximately 36.9microg x kg(-1). In ancient paddy soil, the concentrations of 2-ring and 3-ring PAHs took a larger portion of 63 percents to the total PAHs, and naphthalene and phenanthrene were the most abundant compounds, while PAHs of more than 4 rings took a small part. The ratios of phenanthrene anthracene and benzo(a) anthracene chrysene and 13C-NMR spectrum of soil organic matter showed that PAHs in ancient paddy soil mainly originated from rice straw burning, biogenesis under reducing conditions may be another source.

Aim: To study the pathogenetic processes and the role of gene expression by microarray analyses in expediting our understanding of the molecular pathophysiology of pancreatic adenocarcinoma, and to identify the novel cancer-associated genes., Methods: Nine histologically defined pancreatic head adenocarcinoma specimens associated with clinical data were studied. Total RNA and mRNA were isolated and labeled by reverse transcription reaction with Cy5 and Cy3 for cDNA probe. The cDNA microarrays that represent a set of 4 096 human genes were hybridized with labeled cDNA probe and screened for molecular profiling analyses., Results: Using this methodology, 184 genes were screened out for differences in gene expression level after nine couples of hybridizations. Of the 184 genes, 87 were upregulated and 97 downregulated, including 11 novel human genes. In pancreatic adenocarcinoma tissue, several invasion and metastasis related genes showed their high expression levels, suggesting that poor prognosis of pancreatic adenocarcinoma might have a solid molecular biological basis., Conclusion: The application of cDNA microarray technique for analysis of gene expression patterns is a powerful strategy to identify novel cancer-associated genes, and to rapidly explore their role in clinical pancreatic adenocarcinoma. Microarray profiles provide us new insights into the carcinogenesis and invasive process of pancreatic adenocarcinoma. Our results suggest that a highly organized and structured process of tumor invasion exists in the pancreas.

Objective: To clarify the clinicopathological significance of lymphatic vessel density (LVD) and distribution in pancreatic cancer., Methods: We measured LVD in 43 pancreatic cancer specimens by immunostaining with specific lymphatic endothelium marker, and examined their relationship with well-defined clinicopathological variables., Results: Intratumoral LVD (9.4 +/- 10.0) was significantly lower than periturmoral (16.0 +/- 9.7) (P < 0.001) and nontumoral LVD (13.5 +/- 6.0) (P < 0.01). Increased peritumoral LVD correlated significantly with tumor staging (P < 0.05) and lymph node involvement (P < 0.05)., Conclusion: The lymphatic vessels distribution in pancreatic cancer samples and peritumoral lymphangiogenesis may promote the malignant progression and lymph node metastasis of pancreatic cancer.

A pot culture experiment was carried out to study the influence of arbuscular mycorrhizal (AM) fungi on the growth and Cu uptake of E. splendens. Five levels of Cu (0, 50, 100, 200, 400mg x kg(-1)) and two AM fungal inocula, Glomus caledonium 90036 (36) and Acaulospora mellea ZZ (ZZ), and one nonmycorrhizal inoculum (CK), were applied to the soil under sterilized conditions. The plants were harvested after 60 days of growth. Although decreasing with the increase of Cu levels, root infection rates of all the mycorrhizal plants were >50%, which showed that AM fungi colonized E. splendens roots easily. Shoot and root dry matter yields were all increased by two AM inocula at all Cu levels. Shoot Cu concentrations in the plants treated with two AM inocula increased at 200mg x kg(-1) Cu level and below, but did not change significantly at 400mg x kg(-1) Cu level. Root Cu concentrations in the plants treated with two AM inocula did not change at 0 mg x kg(-1) Cu level, but decreased at 50 mg x kg(-1) Cu level and above. In comparison with nonmycorrhizal plants, shoot Cu uptake by mycorrhizal plants all increased at any Cu level, while root Cu uptake increased only at 100 mg x kg(-1) Cu level. In general, AM fungal inoculation can increase Cu translocation from roots to shoots and shoot Cu uptake by E. splendens.

Objective: To observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells., Methods: SDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control., Results: The level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively., Conclusion: Down-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.

Objective: To determine the clinicopathologic characteristics and the relationship between related gene expression and pathobiologic behavior of pancreatic mucinous noncystic adenocarcinoma., Methods: Among the 249 pancreatic carcinoma cases from the department files, 6 tumors were identified to meet the pathologic criteria of colloid carcinoma. Envision immunohistochemical staining technique was used to detect expression of p21(ras), p21(WAF1), p16, p33(ING1), p53, ATM, MDM2, PCNA, Cyclins (D1, D3, A, B and E). Intra- and extra- cellular mucin production were determined by AB-PAS staining. Clinically, all of 6 cases were followed to June, 2003., Results: In all 6 cases, the tumors were located in the head of the pancreas and all displayed similar microscopic findings. Duodenal invasion was seen in 4 cases and perineural invasion was seen in 1 case. Tumor metastasis in the liver was seen in 2 cases and in the regional lymph nodes in 2 cases. Positive immunostaining was seen in 5 cases with p21(ras), 3 cases with p21(WAF1), 1 case with p16, 4 cases with p33(ING1), 2 cases with p53, 3 cases with ATM, 3 cases with MDM2, 6 cases with PCNA, 3 cases with cyclinA, 3 cases with cyclinD1, 4 cases with cyclinD3, 4 cases with cyclinB and 6 cases with cyclinE. Both extracellular and intracellular mucin was strongly positive for AB-PAS staining. Clinical follow-up found that 2 patients died of their tumors at 14 and 20 months. Three patients were alive after 28, 49 and 87 months of follow-up. One case were lost contact., Conclusions: Pancreatic mucinous noncystic adenocarcinoma has distinct morphologic features and biologic behavior. Multiple gene products including many cyclins may be involved in the pathogenesis of pancreatic colloid carcinoma. The tumor has an aggressive behavior with a high frequency of invasion and metastases, though the prognosis could be better than that of ordinary ductal adenocarcinoma of pancreas.

Objective: To study the effects of RNA interference inhibiting stromal cell derived factor-1 (SDF-1) expression on the proliferation and apoptosis of co-cultured Jurkat cells., Method: Inhibition of SDF-1 expression by RNA interference (RNAi) was achieved by transferring SDF-1 specific short hairpin RNA (shRNA) expressing plasmid into cultured human acute leukemic bone marrow stromal cells. Resistant clones were obtained by G418 selection (group A). The concentration of SDF-1 protein in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The population double time (PDT), cell cycles, apoptosis rates and the expressions of PCNA, Bcl-2/Bax, Fas/FasL of co-cultured Jurkat cells were detected by cells counting, flow cytometry. TdT-mediated dUTP nick-end labelling (TUNEL) and immunocytochemistry (ICC), respectively. The un-transfected acute leukemic (group B) and normal (group C) bone marrow stromal cells were taken as controls., Results: The content of SDF-1 protein in supernatant of group A\[(384 +/- 41) pg/ml] was significantly lower than that in group B[(2474 +/- 271) pg/ml] or group C[(1324 +/- 154) pg/ml]. As group A compared with group B and group C, the PDT of co-cultured Jurkat cells was prolonged (group A: 42 h, vs group B: 29 h, group C: 33 h), and G(0)/G(1) stage cells increased [group A: (28.47 +/- 2.39)%, vs group B: (19.43 +/- 2.80)%, group C: (27.15 +/- 2.07)%], S stage cells decreased [group A: (25.57 +/- 1.90)%, vs group B: (74.48 +/- 3.23)%, group C: (60.99 +/- 2.33)%], G(2)/M stage cells increased [group A: (45.96 +/- 3.24)%, vs group B: (6.09 +/- 1.96)%, group C: (11.86 +/- 1.98)%], the apoptosis rate increased [group A: (15.2 +/- 0.8)%, vs group B: (5.4 +/- 0.7)%, group C: (9.5 +/- 0.4)%], and the expressions of PCNA, Bcl-2, Fas decreased; whereas the expressions of Bax and FasL were increased., Conclusion: The inhibition of SDF-1 expression in bone marrow stromal cells inhibits the proliferation and promotes the apoptosis of co-cultured Jurkat cells.

Objective: To analyze the expression of stromal cell derived factor-1 (SDF-1) and its functional chemokine receptor CXCR4 in patients with acute leukemia and malignant lymphoma., Methods: CXCR4 expressed on the membrane surface of marrow mononuclear cells was enumerated with 3-color flow cytometry. The serum level of SDF-1 was determined with ELISA assay., Results: The expression of SDF-1 and CXCR4 in acute leukemia for ALL group, the expression of SDF-1 and CXCR4 is (7115.8 +/- 946.5) ng/L and (77.2 +/- 9.7)%; for ANLL group, the expression is (4642.2 +/- 1146.8) ng/L, (38.9 +/- 11.0)% and malignant lymphoma for HD group, the expression of SDF-1 and CXCR4 is (3728.9 +/- 690.9) ng/L and (9.2 +/- 2.7)%; for NHL group, the expression is (4442.1 +/- 1073.0) ng/L and (8.5 +/- 2.4)% patients were higher than that in controls the expression of SDF-1 and CXCR4 is (2369.3 +/- 966.5) ng/L and (2.7 +/- 1.5)% (P < 0.01). In the leukemia group, the patients with extra-marrow infiltration have higher expression of SDF-1/CXCR4 than those without and the patients; in the lymphoma group., Conclusions: The high expression of SDF-1 and CXCR4 is somewhat correlated with the pathogenetic process and infiltration characteristics of acute leukemia and malignant lymphoma. This can provide a new method to cure the above-mentioned hematologic tumors.

Background & Objective: Tumor microenvironment affects tumor cells growth. Bone marrow microenvironment may protect leukemic cells from drug-induced damages, but the mechanism is unclear. This study was to explore the protection of bone marrow stromal cells (BMSCs) on leukemic cells against chemotherapy and its mechanism., Methods: Normal and leukemic BMSCs were isolated using Percoll, and cocultured with human acute lymphocyte leukemic cell line Jurkat cells in vitro. After treatment of 0.5 micromol/L of daunorubicin (DNR), apoptosis and cell cycle distribution of Jurkat cells were analyzed by flow cytometry., Results: When treated with DNR for 24 h, apoptosis rate of normal BMSCs-cocultured Jurkat cells was significantly lower than that of Jurkat cells without coculture [(8.39+/-4.08)% vs. (16.02+/-1.00)%, P < 0.05], and apoptosis rate of leukemic BMSCs-cocultured Jurkat cells was significantly lower than that of normal BMSCs-cocultured Jurkat cells [(5.73+/-1.78)% vs. (8.39+/-4.08)%, P < 0.05]; G(0)/G(1) phase percentage of BMSCs-cocultured Jurkat cells was significantly higher than that of Jurkat cells without coculture (P < 0.05), but the difference between Jurkat cells cocultured with normal and leukemic BMSCs was not significant (P > 0.05)., Conclusion: Leukemic BMSCs may inhibit DNR-induced apoptosis in leukemic cells partly through G(0)/G(1) phase arrest.

This study was aimed to explore the influence of anti-CXCR4 monoclonal antibody 12G5 on killing effect of cytosine arabinoside (Ara C) to HL-60 cell, and to assess its therapeutic value in marrow residual disease. HL-60 cells were cultured and co-cultured with leukemic stromal cells, and SDF-1 activity was inhibited with 10 microg/ml 12G5, then, killing effects of Ara C on HL-60 cells were investigated by MTT and morphology assay. Curves by MTT assay revealed that in the test group of 20 microg/ml Ara C, A(540) values decreased slowly but straightly, however, in control group A(540) values decreased markedly for the first two days, and increased from day 3 or 4. In the test group of 40 microg/ml Ara C, although increasing at constricted range of 7 - 9 days, A(540) values decreased in whole observing period of 12 days, while in control group A(540) values decreased markedly at day 0-3, and increased from day 4. Furthermore, two curves go across each other at day 5, and continue the increasing tendency. Morphology results showed that in both treated groups, the number of HL-60 cell decreased markedly and increased gradually in control group, but just contrary to test group. It is concluded that 12G5 may weaken the killing effect of Ara C on HL60 cell in earlier period, but reinforce the total killing effect and delay the occurrence of drug resistance simultaneously. Thus 12G5 has the therapeutic potential on marrow residual disease.

The aim was to analyze the expression level of stromal cell derived factor-1 (SDF-1) and its functional chemokine receptor CXCR4 in the patients with hematologic malignant tumor and their clinic significance. 28 patients with hematologic malignant tumor and 12 normal controls were chosen to be experimental objects. CXCR4 expressed on the cell membrane in bone marrow was enumerated by flow cytometry and serum level of SDF-1 was determined by ELISA assay. The result showed that the expression of SDF-1 and CXCR4 in hematologic malignant tumor were higher than that in normal controls, and the expression levels of two molecules were correlated. What is more, the different hematologic malignant tumor had different CXCR4 expression. In conclusion, the high expression of SDF-1 and CXCR4 in serum and bone marrow cells can be used as detective factors to hematologic malignant tumor. A correlation exists between the high expression of CXCR4 and the infiltration of hematologic malignant cells.

To study the possibility of separation and culture of human umbilical cord blood adherent cell (HUCBAC), the umbilical cord blood CD34(+) cells were cultured in Dexter system in order to evaluate and observe the biological behavior of adherent cells in vitro. The results showed that all cells were cultured with Dexter system. By day 9-14 (at a median of 11.2 days), adherent cell colonies formed and reached their maximum at 15-22 days (mean 19.6 days), by day 28, all adherent cells spread over the bottom of Petri dish. By means of light microscopy, these cells were found to differentiate into three kinds of cells in culture of 28 days: fibroblast-liked cell, macrophage liked cell and small-round cells. The ratio of these three kinds of cells was 56.8%, 38%, 5.5% respectively. Cytochemistry assay revealed that the positive rate reached 100% in NSE stain and PAS stain; the adherent cell by ALP stain were shown 35% positive, but in POX stain the result was negative. Immunohistochemistry stain revealed that the positive rate of cord adherent cells for CD106, CD29, CD44, CD45, CD50, Fn, Ln, collagen IV etc reached 96%, 93%, 98%, 68%, 72%, 92%, 74%, 83% respectively. It is concluded there are hematopoietic adherent precursors in cord blood CD34(+) cells and the HUCBAC shows some biological behavior of hematopoietic stromal cells.

Background & Objective: Stromal cell derived factor-1 (SDF-1), excreted by bone marrow stromal cells, may be involved in shielding of marrow stromal cells from leukemia cells by binding to its receptor CXCR4, but the relation between SDF-1/CXCR4 axis and leukemia cells is unclear. This study was to inhibit activity of SDF-1 by anti-CXCR4 monoclonal antibody 12G5, observe changes of adhesion and proliferation of acute myelocytic leukemia cell line HL-60 co-cultivated with leukemic marrow stromal cells, and to assay potential of inhibiting SDF-1-driven processes on treating marrow residual disease., Methods: HL-60 cells were co-cultured with leukemic marrow stromal cells, and 10 mug/ml 12G5 was added to block SDF-1 activity. Adhesive state, adhesion rate, apoptosis rate, and cell cycle of HL-60 cells were observed after incubation. Living conditions of HL-60 cells were detected by trypan blue exclusion, cell growth curve was protracted., Results: Adhesion rate of 24-h 12G5-incubated HL-60 cells was (39.4+/-7.9)%, while that of control HL-60 cells was (51.4+/-5.9)% (P< 0.05). G0/G1, S, and G2/M phases of 24-h 12G5-incubated HL-60 cells were (55.2+/-4.9)%, (30.4+/-4.1)%, and (14.4+/-5.2)%, respectively, and apoptosis rate was (9.0+/-1.7)%; while those of control HL-60 cells were (44.7+/-2.2)%, (45.3+/-3.7)%, (10.0+/-2.6)%, and (4.0+/-2.4)%, respectively. T test showed that 12G5 induced more cells entering G0/G1 phase (P< 0.05), and increased cell apoptosis rate(P< 0.05). Compared with control HL-60 cells, survival rate and proliferation of 48-h 12G5-incubated HL-60 cells decreased markedly., Conclusion: 12G5 may inhibit adhesion ability and proliferation of HL-60 cells, thus inhibiting SDF-1 activity by 12G5 may be helpful to the treatment of marrow residual disease.

To study the importance of chemokine SDF-1 in surviving of acute myelocytic leukemia cells HL-60, the adhesion ability of HL-60 and expression of Bcl-2, Fas protein when SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody (12G5) were compared with those detected before MAb incubation, in this experiment, HL-60 cell were cultured and co-cultured with normal marrow stromal cell. The adhesion rate was detected while the expression of Bcl-2 and Fas proteins were assayed by immunohistochemical technique when SDF-1 activity was inhibited. The results showed that cell adhesion rate of HL-60 decreased while the expression Bcl-2 decreased and Fas increased. It is concluded that inhibition of SDF-1 activity increases cell apoptosis and thus reduces life-span of leukemia cell at certain level.

This study was aimed to investigate the importance of chemokine SDF-1 in maintaining proliferation ability of acute myelocytic leukemia cell line HL-60 when the effects of SDF-1 on HL-60 cell proliferation were inhibited. Marrow stromal cells were cultured and co-cultured with HL-60 cells, and SDF-1 activity was blocked with anti-CXCR4 McAb. HL-60 cell activity was detected by MTT while cell cycle and the expression of CXCR4 on HL-60 cell membrane were observed by flow cytometry meanwhile. The internal calcium ionic concentration in HL-60 cell was detected as well before and after treated with 12G5. The results showed that 12G5 down-regulated the expression of CXCR4 on HL-60 cell membrane; HL-60 cells at G(0)/G(1) phase increased, but decreased at S phase; survive rate of leukemia cells reduced; the intercellular calcium ionic concentration of HL-60 cell decreased after treated with 12G5. It was concluded that brockage of the SDF-1 activity may inhibit proliferation of leukemia cell at certain level.

A survey was made of the arbuscular mycorrhizal (AM) status of five dominant wild plants Tamarix chinensis, Phragmites communis, Suaeda glauca, Aeluropus littoralis var. sinensis and Cirsium setosum in saline-alkaline soils of the Yellow River Delta that show low plant diversity. All of the species were colonized and showed typical AM structures (arbuscules, vesicles). The colonization percentage ranged from 0.2% to 9.5%, where C. setosum was the highest. The species richness of AMF at the different sites ranged from 2.00 to 2.40 per 50 ml soil, with an average of 2.16. Species diversity ranged from 1.99 to 2.22 per 50 ml soil, with an average of 2.13. Spore density ranged from 3 to 30 per 50 ml soil, with an average of 12. Glomus was the dominant genus, with a frequency and relative abundance of 88.1% and 68.4%, respectively. G. caledonium, with a frequency and relative abundance of 15.0% and 4.6%, respectively, was the dominant species. Differences were also observed in the distribution of AMF in different soil layers. Although there were still AM fungal spores in the layer 40 cm below the surface, most spores were found at a depth of 0-40 cm.

To study the role of hematopoietic microenvironment abnormality in development of minimal residual disease and its mechanism, the viability of HL-60 cells was investigated by means of bone marrow stromal cell culture system or co-culture system of bone marrow stromal cell with HL-60 cells and idarubicin (IDA), flow cytometry and ELISA. The results showed that viability of HL-60 cells gradually decreased along with the increase of IDA dose and prolongation of culture time. Amount of HL-60 cells co-cultured with leukemia bone marrow stramal cells was significantly increased as compared with that of the control (P < 0.05). Bone marrow stromal cells or stromal cell conditioned medium reduced the effect of IDA on HL-60 cells in culture. In conclusion, leukemia bone marrow stromal cells contribute to increasing resistance of HL-60 cells to chemotherapeutic agents, and play some role in developing minimal residual disease.

The effect of inoculation with arbuscular mycorrhizal (AM) fungi (Acaulospora lavis) on the degradation of di(2-ethylhexyl) phthalate (DEHP) in soil was studies. Cowpea plants (Pigna sinensis) were used as host plants and grown in a specially designed rhizobox. The experimental results indicated that, both in sterile and non-sterile soil, mycorrhizal colonization rates were much higher in the mycorrhizal plants than in the non-mycorrhizal plants. Addition of 4 mg/kg DEHP slightly affected mycorrhizal colonization, but the addition of 100 mg/kg DEHP significantly decreased mycorrhizal colonization. DEHP degradation in the mycorrhizosphere (Ms) and hyphosphere (Hs), especially in the Hs, increased after inoculation with Acaulospora lavis. It is concluded that mycorrhizal hyphae play an important role in the plant uptake, degradation and translocation of DEHP. The mechanism might be attributed to increased numbers of bacteria and actinomycetes and activity of dehydrogenase, urease and acid phosphatase in the Ms and Hs by mycorrhizal fungi.

Objective: To analyse the clinical features of uncinate process carcinoma of the pancreas and the diagnosis and treatment of this malignancy., Method: Fifty-nine patients with pancreas uncinate process carcinoma treated from January 1998 to September 2002 at our hospital were analysed retrospectively., Results: Major symptoms of these patients were upper abdominal pain accompanied with lumbar pain, body weight loss and jaundice. Thirty-seven patients received regional pancreaticoduodenectomy (RP), 16 partial resection of the superior mesenteric vein-portal vein (SMV-PV) or superior mesenteric artery (SMA) and reconstruction, 1 anhydrous alcohol injection in the celiac nerve plexus, regional chemotherapy via a chemotherapy pump, and liver biopsy, and 5 no operation. The survival of the patients after operation was 2-46 months (median 12.1 months). Eleven patients are still alive with a longest survival of 46 months. The 1- and 3-year survival rates were 37.7% and 5.6%., Conclusions: Pancreas uncinate process carcinoma invading the adjacent SMV/SMA-PV causes difficulty in early diagnosis and poor prognosis, which are related to its location, not tumor's aggressive nature. This carcinoma has a high resection rate of 89.8%.

Objective: To prospectively evaluate the long-term effect of pancreaticoduodenectomy with regional lymphadenectomy., Methods: One hundred and twenty-one patients with ductal adenocarcinoma in the pancreatic head treated from 1996 to 2001 were studied prospectively. The enrollment of the patients was dependent on 7 criteria. The patients were divided into two groups: regional lymphadenectomy (group A, n = 50) and routine Whipple procedure (group B, n = 71). Their pre- and postoperative conditions, clinicopathological data, survival rates were studied., Results: It was comparable between the 2 groups in age, sex, preoperative risk factors, operative management, and postoperative complication. Clinicopathological results showed no difference in tumor size and plexus invasion; but the frequency of lymph node involvement and the amount of resected lymph node in group A were significantly higher than those in group B. The rate of local recurrence was significantly higher in group A than in group B. The survival rates of 1-, 3-, 5-year in group A were 70.8%, 31.4%, 20.9%, respectively, which were higher than those in group B. No direct relations were observed between nodal involvement and survival rate., Conclusion: Lymphadenectomy in radical pancreaticoduodenectomy could remove lymph nodes effectively and sufficiently, and reduce the rate of local recurrence so as to improve the long-term survival rate.

Aim: To identify new diagnostic markers and drug targets, the gene expression profiles of pancreatic cancer were compared with that of adjacent normal tissues utilizing cDNA microarray analysis., Methods: cDNA probes were prepared by labeling mRNA from samples of six pancreatic carcinoma tissues with Cy5-dUTP and mRNA from adjacent normal tissues with Cy3-dUTP respectively through reverse transcription. The mixed probes of each sample were then hybridized with 12 800 cDNA arrays (12 648 unique human cDNA sequences), and the fluorescent signals were scanned by ScanArray 3 000 scanner (General Scanning, Inc.). The values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated by ImaGene 3.0 software (BioDiscovery, Inc.). Differentially expressed genes were screened according to the criterion that the absolute value of natural logarithm of the ratio of Cy5-dUTP to Cy3-dUTP was greater-than 0.69., Results: Among 6 samples investigated, 301 genes, which accounted for 2.38 % of genes on the microarry slides, exhibited differentially expression at least in 5. There were 166 over-expressed genes including 136 having been registered in Genebank, and 135 under-expressed genes including 79 in Genebank in cancerous tissues., Conclusion: Microarray analysis may provide invaluable information on disease pathology, progression, resistance to treatment, and response to cellular microenvironments of pancreatic carcinoma and ultimately may lead to improving early diagnosis and discovering innovative therapeutic approaches for cancer.