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The application of microbial fertilizer plays an important role in improving soil restoration and fertilizer utilization. The effects of microbial fertilizer are greatly affected by crop genotypes and ecological conditions. Little is known about the effects of microbial fertilizers on maize production in Northeast China. To develop microbial fertilizer specific to the black soil and the climate characteristics of Northeast China, we isolated five plant rhizosphere-promoting bacteria (PGPR), named as MZ1, MZ2, MZ3, MZ4 and MZ5, with different degrees of biological functions such as IAA synthesis, phosphate-solubilizing, potassium-solubilizing and siderophore-releasing, from the rhizosphere of maize field. The analysis of ecological adaptability showed that those five strains differed in salt resistance, drought tolerance, acid and alkali resistance, pesticide resistance. The 16S rRNA gene sequences analysis showed that the strains MZ1, MZ2, MZ3, MZ4 and MZ5 belonged to the genus of Sphingomonas, Enterobacter, Pseudomonas, Bacillus and Rhizobium, respectively. In maize field experiment with 50% nitrogen fertilizer reduction, the inoculation with MZ1, MZ3 and MZ5 increased grain yield by 19.9%-25.0%. MZ1, MZ3, and MZ5 could be used as microbial fertilizers for maize in Northeast China.

This study was aimed to compare the expressions of specific transcription factors of CD4(+) T cell subset ( T-bet, GATA-3, RORγt and FoxP3 mRNA) in peripheral blood of patients with aplastic anemia(AA), myelodysplastic syndrome(MDS), and acute myeloid leukemia(AML), and investigate their immune status and pathogenesis, so as to provide experimental basis for the choice of clinical treatment. The expression of T-box (T-bet), GATA-3, ROR-γt and Foxp3 mRNA in PBMNC were examined by RT-PCR in 42 cases of MDS, including 22 refractory anemia(MDS-RA) and 20 refractory anemia with excess blasts (MDS-RAEB), in 23 cases of AA, 17 cases of AML patients and 16 healthy volunteers respectively. The results indicated that, compared with normal control group, expressions of T-bet and RORγt mRNA in AA patient group were significantly higher (P < 0.01), expression levels of GATA3 Foxp3 mRNA were lower (both P < 0.01). There was no significant difference in expression of T-bet and GATA3 mRNA between MDS group and normal control group, but the expression levels of Foxp3 and RORγt mRNA were higher than those in normal controls (P < 0.05); T-bet and RORγt in MDS-RA group were higher than those in the normal controls(P < 0.01), and GATA3 expression significantly reduced (P < 0.05), however, there was no significant difference in expression of Foxp3 between MDS-RA and the controls. Expression levels of T-bet and RORγt mRNA in patients with MDS-RAEB and AML were lower than those in normal controls (P < 0.05), but the expression levels of GATA3 and Foxp3 mRNA were significantly higher than those in normal controls (P < 0.01). It is concluded that the transcription factor expressions are different in PBMNC of patients among these three diseases. Immune-mediated excessive apoptosis may play an important role in pathogenesis, bone marrow failure in patients with AA and MDS-RA, and abnormal clones of immature cells may be one of main reasons for bone marrow failure in AML and late stage of MDS.

The aim of this study was to detect the mRNA expression of tissue factor pathway inhibitor-2 ( TFPI-2) and its methylation in bone marrow mononuclear cells from acute myeloid leukemia (AML) patients and to explore its significance in AML. Bone marrow mononuclear cells were isolated from newly diagnosed AML patients (n = 33), complete remission AML patients (n = 19), relapsed/refractory AML patients (n = 12) and iron deficiency anemia patients (control group, n = 15). Expression of TFPI-2 mRNA was detected with real-time quantitative PCR (RT-PCR) and the methylation of CpG island in its promoter was detected with methylation-specific PCR (MSP). The results showed that the expression of TFPI-2 mRNA in newly diagnosed AML, complete remission AML and relapsed/refractory AML patients was much lower than that in the controls (P < 0.05). Furthermore, its expression in relapsed/refractory AML patients was lower than that in newly diagnosed AML patients (P = 0.006). Compared with complete remission AML patients, the expression of TFPI-2 mRNA in newly diagnosed AML patients was significantly reduced (P = 0.030). The percentage of TFPI-2 promoter methylation in AML patients was 64.63% (42/64). In newly diagnosed AML group, complete remission AML group and relapsed/refractory AML group,the percentages of TFPI-2 promoter methylation were 66.67% (22/33), 52.63% (10/19) and 83.33% (10/12) (P > 0.05), respectively. The optical density ratio of TFPI-2 mRNA expression was 0.165 (0.005-2.099) in methylated AML patients, and 0.597 (0.011-2.787) in unmethylated AML patients (P < 0.05). Methylation of TFPI-2 gene promoter was not detected in control patients. After 2 courses of chemotherapy, the level of TFPI-2 mRNA was much higher in the CR group than that in the non-CR group (P < 0.05). It is concluded that the down-regulation or silence of TFPI-2 gene potentially results from its promoter methylation, and the expression level of TFPI-2 and the methylation status of its promoter may be used as indicators of risk stratification and evaluation of disease progress.

Objective: To investigate the role of mast cells and interleukin-9 (IL-9) in B-cell non-Hodgkin lymphoma (B-NHL) development and its clinical significance., Methods: The expression level of CD117 in tumor tissues of 32 B-NHL patients was determined by Western blot. The infiltration of CD117⁺ mast cells (MCs) in human B-NHL tumor tissues was observed by immunohistochemistry staining. To evaluate the correlations between the data from CD117⁺ MCs and biological markers of human B-NHL, a Spearman correlation coefficient (rs) was calculated. IL-9 levels in sera of B-NHL patients were measured by ELISA. Effects of IL-9 on expressions of functional genes of mouse bone marrow-derived mast cells (BMMCs) were detected by real-time quantitative RT-PCR., Results: The expression of CD117 was upregulated significantly in human B-cell NHL involved tissues when compared with that of controls (0.0551±0.0064 vs 0.0192±0.0072, P<0.01). Infiltration of more CD117⁺ MCs was found in tissues from B-cell NHL subjects compared with that of controls. IL-9 level in serum samples from patients with B-cell NHL was higher than that from healthy controls. Addition of rIL-9 to the culture gave rise to increase in the purity of mouse BMMCs in the first three weeks. In vitro culture experiments showed that the addition of IL-9 could induce the differentiation of mouse BMMC and the expressions of MC-related genes, including CD117, Fcer1α, Mcpt1 and Mcpt5., Conclusion: Our study showed that IL-9 promoted immune response mediated by MCs, and probably played important roles in B-NHL growth. Pharmacological or targeted inhibition of mast cells or IL-9 activity may provide new strategy for B-cell NHL therapy.

This study was purposed to detect the balance and the activity change of cytotoxic T cell subsets in aplastic anemia (AA) patients, myelodysplastic syndrome (MDS) patients and acute myeloid leukemia (AML) patients, and to explore the cellular immune mechanism for abnormal hematopoiesis of the three diseases, so as to provide experimental basis for the choice of clinical treatment. The proportion of the cytotoxic T cells and part of the T-cells subsets in peripheral blood were detected by flow cytometry in 35 cases of MDS, including 19 refractory anemia (MDS-RA), 16 refractory anemia with excess blasts (MDS-RAEB), 17 AA, 15 AML patients and 10 normal donors respectively. The results showed that compared with the control group, the percentage of Tc1, Tc1/Tc2, CD8(+)HLA-DR(+), CD3(+)CD8(+)CD28(+), CD8(+)CD45RO(+) cells was significantly higher and the percentage of CD8(+)CD45RA(+) was significantly lower in AA and MDS-RA group. There was no difference in the percentage of Tc2 cells between AA/MDS-RA and normal controls; the percentage of CD8(+)CD45RO(+) cells was significantly higher and the percentage of Tc1, CD3(+)CD8(+)CD28(+), CD8(+)HLA-DR(+) was significantly lower in MDS-RAEB group, the percentage of CD8(+)CD45RA(+) was lower but the difference was not significant, and there was no difference in the percentage of Tc, Tc1/Tc2 cells between MDS-RAEB group and the control group. The percentage of Tc2 cells was significantly higher and the percentage of other parameters was significantly lower in AML group than those of normal controls. It is concluded that the cellular immune statuses in AA, the different stages of MDS and AML are different. In AA and the early stage of MDS, the balance of Tc1/Tc2 shifts to Tc1, and the activation of T-cell subsets increases. In the late stage of MDS and AML, the balance of Tc1/Tc2 shifts to Tc2, the activation of T-cell subsets decreases. The former may be closely related to bone marrow failure while the latter may be one of the important mechanisms in which the malignant clones escape from immune effect.

Fire is one of the important natural disturbances to forest ecosystem, giving strong impact on the ecosystem carbon dynamics. By using CENTURY model, this paper simulated the responses of the carbon budget of Larix gmelinii forest in Huzhong area of Daxing' an Mountains to different intensities of fire. The results indicated that after the fires happened, the soil total carbon pool of the forest had a slight increase in the first few years and then recovered gradually, while the stand biomass carbon pool increased after an initial decrease, with the recovery rate of carbon pool of the stand fine components being faster than that of the coarse components. The fluctuation of the carbon pools increased with the increase of fire intensity. After the fires, both the net primary productivity (NPP) of forest vegetation and the soil heterotrophic respiration increased after an initial decrease, but the recovery rate of the NPP was faster than that of soil heterotrophic respiration, resulting in the alternation of the stand functioned as a carbon source or sink. After light fire, the forest still functioned as a weak carbon sink, and quickly recovered as a carbon sink to the level before the fire happened. After other intensities fire, the forest functioned as a carbon source within 9-12 years, and then turned back to a carbon sink again. It was suggested that lower intensity forest fire could promote the regeneration of L. gmelinii forest, reduce the combustibles, and have no strong impact on the stand carbon budget, while higher intensity forest fire would lead to the serious loss of soil- and tree carbon sequestration, retard the recovery of the forest, and thereby, the forest would be a carbon source in a longer term.

Objective: To investigate the role of IL-18 and IL-18BP balance in aplastic anemia (AA)., Methods: A total of 29 AA patients and 22 controls were recruited in present research. The expressions of IL-18 and IL-18BP were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of IL-18 and IL-18BP were measured in all subjects using real-time quantitative polymerase chain reaction (RT-PCR)., Results: The levels of the IL-18 in plasma of AA and normals were (365.5 ± 160.6) pg/ml and (175.9 ± 92.8) pg/ml (P < 0.01); and the expression of IL-18 in severe AA patients (441.3 ± 116.9) pg/ml were higher than that in non-severe AA patients (326.4 ± 167.0) pg/ml (P < 0.05). The level of IL-18BP was increased in plasma of AA (1788.6 ± 523.8) pg/ml than in normals (1083.6 ± 489.6) pg/ml (P < 0.05). But the ratio of IL-18/IL-18BP in AA patients was much higher than that in controls (P < 0.05). RT-PCR revealed the levels of IL-18 and IL-18BP mRNA were up-regulated in AA patients when compared to controls, but the ratio of IL-18/IL-18BP was significantly elevated in AA patients., Conclusion: IL-18/IL-18BP imbalance may play an important role in pathogenesis of AA and regulating the balance of IL-18 and IL-18BP may be a therapeutic approach to AA.

This study was aimed to investigate the expression and clinical significance of IL-18, IL-18 binding protein (IL-18BP), IFN-γ and IL-4 secreted from splenocytes of patients with idiopathic thrombocytopenic purpura (ITP) in vitro. Spleen mononuclear cells (MNC) were prepared by using routine sterile method, and were cultured in RPMI 1640 complete medium containing 10 µg/ml PHA, 10% fetal calf serum at 37°C and 5% CO2. The levels of IFN-γ, IL-4, IL-18 and IL-18BP secreted from MNC of ITP patients and normal controls were determined after culture for 48 hours. The results showed that after culture of spleen MNC for 48 hours, the levels of IL-18 and IFN-γ were significantly higher in patients with ITP than that in controls, but the levels of IL-18BP was not significantly elevated in ITP patients. The level of IL-4 was below the detectable limit of the assay used. It is concluded that imbalance between IL-18 and IL-18BP may play an important role in pathogenesis of ITP, and regulation of balance between IL-18 and IL-18BP may be a therapeutic approach against ITP.

In this study, two phenanthrene-utilizing and high-effective P-accumulating bacteria Y11 and Y4-2 were isolated from sludge samples of Taihu Lake using plate culture and blue-colored colonies methods. Strains Y11 and Y4-2 were identified as Acinetobacter sp. according to the results of morphology, physiology and the phylogenetical analyses of 16S rDNA sequences. The two strains showed high effective P-accumulating ability in plate and broth cultures. Y11 could grow well from 10 degrees C to 35 degrees C and pH from 6 to 9, Y4-2 could grow well from 10 degrees C to 35 degrees C and pH from 6 to 8. Phenanthrene could be used as sole carbon and energy sources. In our experiment, high phosphorus concentration in broth culture had no negative effect on the growth of strain Y11, however the growth of strain Y4-2 was slightly affected. Under cultivation condition of 30 degrees C, 170 r/min, 1% inoculation (D600 = 0.4), the accumulation of phosphorus was various by strains and cultivation concentration of phosphorus as well: In 2 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 96.13% and 94.65%, the phosphorus concentration decreased from 2 mg/L to 0.08 mg/L and 0.11 mg/L, respectively. In 5 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 95.94% and 71.19%, the phosphorus concentration decreased from 5 mg/L to 0.20 mg/L and 1.44 mg/L, respectively. In 8 mg/L phosphorus concentration medium, the highest phosphorus-accumulation ratio of strains Y11 and Y4-2 were 71.24% and 47.81%, the phosphorus concentration decreased from 8 mg/L to 2.30 mg/L and 4.18 mg/L, respectively. For the P removal of Yunnan Dianchi water sample containing 1.01 mg/L phosphorus, Y11 and Y4-2 were successful to decrease the phosphorus concentration from 1.01 mg/L to 0.06 mg/L in 6 h and 48 h, respectively. The results indicated that Acinetobacter sp.Y11 and Y4-2 possess high ability of phosphorus removal and suitability to various water environments. Y11 strain adapt to all kinds of eutrophic waterbody even polluted with phenanthrene, Y4-2 adapt to pH < or = 8.0 eutrophic waterbody with phenanthrene pollution.

Objective: To investigate the effects of protein tyrosine kinase (PTK), protein tyrosine phosphatase (PTP) and protein kinase C (PKC) on apoptosis and observe the changes of cytosolic calcium ([Ca(2+)]i) of arsenic trioxide (As2O3) treated human leukemia cells NB4 and cortex neurons., Methods: [Ca(2+)]i of NB4 cells and cortex neurons was probed with Fluo-3/AM, its changes were assayed with laser confocal microscopy in real-time after As2O3 treatment at different concentrations, the effects of PTK and PTP and the activation of PKC on these changes with confocal microscopy and phosphorus radioisotope assay. DNA ladders of NB4 cells and cortex neurons after exposed to As2O3 were observed., Results: As2O3 at 1 micromol/L could remarkably increase the [Ca(2+)]i of NB4 cells but had no effects on neurons. Vanadate, a kind of PTP inhibitor, could promote the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total increase rates at 240 seconds after exposed to As2O3 at different concentrations were (6.5 +/- 2.3)%, (21.7 +/- 2.1)%, (49.2 +/- 2.5)% for NB4 cells, and (6.7 +/- 2.1)%, (19.4 +/- 2.5)%, (52.3 +/- 2.7)% for cortex neurons, respectively. Genistein, a kind of PTK inhibitor, could decrease the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total inhibited rates at 240 seconds after As2O3 treatment at different concentrations were (6.7 +/- 2.9)%, (25.6 +/- 2.5)%, (52.2 +/- 3.5)% for NB4 cells, and (7.8 +/- 3.1)%, (18.1 +/- 2.8)%, (51.3 +/- 3.3)% for cortex neurons, respectively. The activation of PKC began to increase as exposed to As2O3 at 1 micromol/L for 3 h, and kept rising continuously in NB4 cells and at 24 h DNA ladders emerged. However, none of the above results was found in human cortex neurons, but when exposed to 2 micromol/L As2O3, the activation of PKC and DNA ladders did emerge in neurons., Conclusions: The phosphorylation and dephosphorylation of PTK and PTP participated in nonspecific apoptosis signal transduction pathway related to As2O3, and accompanied with PKC activation. The [Ca(2+)]i elevation was closely related to increased PKC activation. There existed difference in dose tolerances to As2O3 between NB4 cell and cortex neurons.

Ion exchanger and ODS bonded silica gel for HPLC packing were prepared. An HPLC method for the analysis of surfactant PS concentration in injected-produced liquor has been established to meet the need of ASP flooding developed in Gudao West Block of Shengli Oilfield, with a minimum detectable limit of 0.4 mg/L, a linear range of 50 mg/L-1,000 mg/L and recoveries of 95.7%-99.8%. This method has provided great technical support in a variety of fields to the design of the prescription for ASP flooding, the regulation of injection measurement, the quality control of products and the study on the mechanism of oil flooding etc.