Bifico Ameliorates Neurological Deficits After Ischemic Stroke in Mice: Transcriptome Profiling.

BACKGROUND/AIM: Evidence suggests that gut microbiota can affect various neurological diseases, including stroke. Stroke patients have an increase in harmful gut bacteria and a decrease in beneficial bacteria. This increases intestinal permeability, increases the risk of infection, and even affects many inflammatory factors. While probiotics may affect stroke prognosis by improving the gut environment. This study aimed to investigate the effect of probiotic Bifico on the neural function in mice after focal cerebral ischemia and explore its mechanisms of action. MATERIALS AND METHODS: A focal cerebral ischemia model was established in mice. Four weeks before modeling, animals were divided into three groups: Stroke plus Vehicle group, Stroke plus Pre-Bifico group and Bifico group. The infarct volume and neurobehaviors were evaluated. Whole-gene expression profiling was performed at different days after treatment (D1, D7, D14, D28) by RNA-seq. Differentially expressed genes (DEGs) were the processed for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). Some inflammation and immune related genes were screened and their expression was analyzed. RESULTS: Compared to the Stroke plus Vehicle group and Bifico group, the infarct volume and neurological score were significantly reduced in the Pre-Bifico group. There were 2 DEGs at D1, 193 DEGs at D7, 70 DEGs at D28 between Stroke plus Pre-Bifico group and Stroke plus Vehicle group. For GO analysis, there were 139 significant terms at D7 and 195 at D28. For KEGG, there were 2 significant pathways at D7 and 9 at D28. Among 87 genes related to inflammation and immunity, 6 DEGs were identified. The expression of CCL9 was significantly elevated at most time points after stroke compared to the Stroke plus Vehicle group, while that of CCL6, CXCL10, CD48, CD72 and CLEC7A was highly expressed only in the recovery stage of stroke. CONCLUSION: Oral pre-treatment with Bifico for 28 days can reduce cerebral infarction and promote recovery of neurological function in stroke mice, which may be ascribed to the regulation of immunity and inflammation in the brain.

Rapid label-free detection of early-stage lung adenocarcinoma and tumor boundary via multiphoton microscopy.

Lung cancer is the most commonly diagnosed cancer and the leading cause of cancer-related deaths in China. Rapid and precise evaluation of tumor tissue during lung cancer surgery can reduce operative time and improve negative-margin assessment, thus increasing disease-free and overall survival rates. This study aimed to explore the potential of label-free multiphoton microscopy (MPM) for imaging adenocarcinoma tissues, detecting histopathological features, and distinguishing between normal and cancerous lung tissues. We showed that second harmonic generation (SHG) signals exhibit significant specificity for collagen fibers, enabling the quantification of collagen features in lung adenocarcinomas. In addition, we developed a collagen score that could be used to distinguish between normal and tumor areas at the tumor boundary, showing good classification performance. Our findings demonstrate that MPM imaging technology combined with an image-based collagen feature extraction method can rapidly and accurately detect early-stage lung adenocarcinoma tissues.

Secondary cell wall patterning-connecting the dots, pits and helices.

All plant cells are encased in primary cell walls that determine plant morphology, but also protect the cells against the environment. Certain cells also produce a secondary wall that supports mechanically demanding processes, such as maintaining plant body stature and water transport inside plants. Both these walls are primarily composed of polysaccharides that are arranged in certain patterns to support cell functions. A key requisite for patterned cell walls is the arrangement of cortical microtubules that may direct the delivery of wall polymers and/or cell wall producing enzymes to certain plasma membrane locations. Microtubules also steer the synthesis of cellulose-the load-bearing structure in cell walls-at the plasma membrane. The organization and behaviour of the microtubule array are thus of fundamental importance to cell wall patterns. These aspects are controlled by the coordinated effort of small GTPases that probably coordinate a Turing's reaction-diffusion mechanism to drive microtubule patterns. Here, we give an overview on how wall patterns form in the water-transporting xylem vessels of plants. We discuss systems that have been used to dissect mechanisms that underpin the xylem wall patterns, emphasizing the VND6 and VND7 inducible systems, and outline challenges that lay ahead in this field.

Dual-wavelength filters based on two-dimensional photonic crystal degenerate modes with a ring dielectric rod inside the defect cavity.

A tunable dual-wavelength filter based on degenerate modes with a ring dielectric rod inside the single point defect cavity is proposed. The band structures and mode profiles are computed by the plane-wave expansion method. The normalized transmission spectra for this structure are investigated by using the two-dimensional finite-difference time-domain method. The two orthogonal output modes of the filter can be regarded as the combination of the original degenerate cavity modes and both excited due to the introduced perturbation rod. The influences of the perturbation rod on the localized modes, band separations, and the tuning ranges are all investigated and analyzed based on the perturbation theory. With the ring dielectric rod being introduced into the defect, a narrower wavelength bandwidth and a wider wavelength tuning range can be obtained, which are superior to that of the filter based on a cavity with a larger radius rod. Besides, the perturbation rod can also be used for splitting degenerate modes for multiwavelength filters. The proposed filter has a simple structure and may be potentially applied in various integrated circuits, such as multiwavelength filters.

Highly nonlinear all-solid photonic crystal fibers with low dispersion slope.

A fluorine-doped trench-assisted structure is proposed to improve the nonlinearity of photonic crystal fibers (PCFs). Three all-solid highly nonlinear PCFs with low dispersion slope and low confinement loss are designed. They exhibit all normal dispersion, two zero dispersion wavelengths (ZDWs) and one ZDW just at 1.55 µm, respectively. The lowest dispersion slope is 5.12×10(-4) ps/(km·nm(2)), which is 2 orders of magnitude lower than that of conventional highly nonlinear fibers. A nonlinear coefficient of 31.5 W(-1)·km(-1) and low loss of 9.62×10(-5) dB/km at 1.55 µm has been achieved for this PCF.

Recovery of copper and water from copper-electroplating wastewater by the combination process of electrolysis and electrodialysis.

In this paper, a laboratory-scale process which combined electrolysis (EL) and electrodialysis (ED) was developed to treat copper-containing wastewater. The feasibility of such process for copper recovery as well as water reuse was determined. Effects of three operating parameters, voltage, initial Cu(2+) concentration and water flux on the recovery of copper and water were investigated and optimized. The results showed that about 82% of copper could be recovered from high concentration wastewater (HCW, >400mg/L) by EL, at the optimal conditions of voltage 2.5 V/cm and water flux 4 L/h; while 50% of diluted water could be recycled from low concentration wastewater (LCW, <200mg/L) by ED, at the optimal conditions of voltage 40 V and water flux 4 L/h. However, because of the limitation of energy consumption (EC), LCW for EL and HCW for ED could not be treated effectively, and the effluent water of EL and concentrated water of ED should be further treated before discharged. Therefore, the combination process of EL and ED was developed to realize the recovery of copper and water simultaneously from both HCW and LCW. The results of the EL-ED process showed that almost 99.5% of copper and 100% of water could be recovered, with the energy consumption of EL ≈ 3 kW h/kg and ED ≈ 2 kW h/m(3). According to SEM and EDX analysis, the purity of recovered copper was as high as 97.9%.

Proliferation and apoptosis of bone marrow CD4(+) T cells in patients with aplastic anemia and impacts of the secreted cytokines on hematopoietic stem cells from umbilical cord blood.

Recent studies indicate that immune-associated aplastic anemia (AA) resembles such autoimmune diseases as insulin-dependent diabetes and chronic autoimmune thyroiditis that belong to organ-specific autoimmune diseases. Many independent investigation groups have successfully isolated the pathopoiesis-associated T cell clone causing hematopoiesis failure with a CD4 phenotype from peripheral blood and bone marrow (BM) in AA patients. In the current study, BM CD4(+) T cells were isolated from AA patients and healthy controls with immunomagnetic beads sorting, and proliferation capability, apoptosis features and the impacts of their secreted cytokines on hematopoiesis stem/progenitor cells were compared between them. By (3)H-TdR method, CD4(+) T cells in AA group presented more enhanced proliferative activity. The stimulation index in control group and AA group was 1.47+/-0.24, and 2.51+/-0.34 respectively (P<0.01). After BM CD4(+) T cells were induced by high concentration of CD3 monoclonal antibody for 18 h, evident apoptosis cells could be seen under the electron microscope in both control group and AA group. Flow cytometry revealed that apoptosis rates in the early and late stages of AA group were significantly higher than in control group (P<0.01). Early-stage apoptosis rate in control and AA groups was (6.85+/-1.48)% and (16.98+/-4.40)%, and late-stage apoptosis rate in control group and AA group was (2.65+/-1.57)% and (7.74+/-0.83)%, respectively (P<0.01). The CFU-GM count in AA group and control group was (74.50+/-9.50)/10(4) cells and (124.25+/-19.80)/10(4) cells respectively under an inverted microscope (P<0.01), and the expression levels of CyclinD3 mRNA and protein in cord blood CD34(+) cells were both down-regulated induced by BM CD4(+) T cell culture supernatant in AA patients. These results indicate that BM CD4(+) T cells of AA patients are likely in an abnormally proliferative, and activated state which can correlate intimately with AA hematopoiesis damage. BM CD4(+) T cells in AA patients can secret some soluble cytokines that can inhibit proliferation of hematopoietic stem cells by suppressing the expression of Cyclin D3, resulting in hematopoiesis failure.

Proliferation and Apoptosis of Bone Marrow CD4+ T Cells in Patients with Aplastic Anemia and Impacts of the Secreted Cytokines on Hematopoietic Stem Cells from Umbilical Cord Blood / 华中科技大学学报（医学）（英德文版）

Recent studies indicate that immune-associated aplastic anemia(AA)resembles such autoimmune diseases as insulin-dependent diabetes and chronic autoimmune thyroiditis that belong to organ-specific autoimmune diseases.Many independent investigation groups have successfully isolated the pathopoiesis-associated T cell clone causing hematopoiesis failure with a CD4 phenotype from peripheral blood and bone marrow(BM)in AA patients.In the current study,BM CD4+ T cells were isolated from AA patients and healthy controls with immunomagnetic beads sorting,and proliferation capability,apoptosis features and the impacts of their secreted cytokines on hematopoiesis stem/progenitor cells were compared between them.By 3H-TdR method,CD4+ T cells in AA group presented more enhanced proliferative activity.The stimulation index in control group and AA group was 1.47±0.24,and 2.51±0.34 respectively(P<0.01).After BM CD4+ T cells were induced by high concentration of CD3 monoclonal antibody for 18h,evident apoptosis cells could be seen under the electron microscope in both control group and AA group.Flow cytometry revealed that apoptosis rates in the early and late stages of AA group were significantly higher than in control group(P<0.01).Early-stage apoptosis rate in control and AA groups was(6.85±1.48)% and(16.98±4.40)%,and late-stage apoptosis rate in control group and AA group was(2.654±1.57)% and(7.74±0.83)%,respectively(P<0.01).The CFU-GM count in AA group and control group was(74.50±9.50)/104 cells and(124.25±19.80)/104 cells respectively under an inverted microscope(P<0.01),and the expression levels of CyclinD3 mRNA and protein in cord blood CD34+ cells were both down-regulated induced by BM CD4+ T cell culture supernatant in AA patients.These results indicate that BM CD4+ T cells of AA patients are likely in an abnormally proliferative,and activated state which can correlate intimately with AA hematopoiesis damage.BM CD4+ T cells in AA patients can secret some soluble cytokines that can inhibit proliferation of hematopoietic stem cells by suppressing the expression of Cyclin D3,resulting in hematopoiesis failure.

[Study on thyroid defunctionalization method for the preoperative preparation of hyperthyroid operation].

OBJECTIVE: To set up and publicize the thyroid defunctionalization method for the preoperative preparation of hyperthyroid operation. METHODS: 476 hyperthyroid patients admitted in our hospital from March 1990 to February 2005 were studied by groups. They were divided randomly into a test group (244 patients), in which "preoperative preparation method of sequential thyroid defunctionalization" was applied to hyperthyroid patients, and based on the different drug dosages and treating terms used, further 4 subgroups (A, B, C and D) were divided to observe the treatment efficiency; And a control group (232 patients), in which antithyroid drugs and iodine preparation were applied preoperatively to cases. Thyroid functions in every stage of treatment were tested by radioimmunoassays (RIA), and operative bleeding volumes and postoperative complications were observed. RESULTS: Compared to the control group, the thyroid congestion and surface varices were alleviated in the test groups, in which the thyroid tissue of subgroup A most closed to euthyroidism histologically. The mean operative bleeding volume of test group was less than that of the control group. The bleeding volumes were (324.76 +/- 163.26) mL for the control group, (195.74 +/- 57.07) mL for the subgroup A, (230.00 +/- 70.81) mL for the subgroup B, (240.47 +/- 80.29) mL for the subgroup C and (314.75 +/- 96.46) mL for the subgroup D. There was no significant difference between the control group and subgroup D, but compared with the subgroup A, B, and C, there was the significant difference between control and treated subgroup (P < 0.05). The postoperative complication rate of the test group was 8.61% (21/244), while that of the control group was 17.24 (40/232). There was statistic difference between two groups (P < 0.005). CONCLUSION: The key to "preoperative preparation method of sequential thyroid defunctionalization" is as follows: the synthesis of thyroxin should be fully inhibited to thyroid defunctionalized; sufficient exogenous thyroxin should be supplemented; the term of thyroid function compensation should be long enough. The "preoperative preparation method of sequential thyroid defunctionalization" can decrease perioperational complications effectively and operation risks.

Screening of aplastic anaemia-related genes in bone marrow CD4+ T cells by suppressive subtractive hybridization.

BACKGROUND: CD4(+) T cells play a crucial role in the pathogenesis of aplastic anaemia. However, the mechanisms of over-proliferation, activation, infiltration of bone marrow and damage to haematopoietic cells of CD4(+) T cells in aplastic anaemia are unclear. Therefore, we screened differentially expressed genes of bone marrow CD4(+) T cells of aplastic anaemia patients and normal donors by suppressive subtractive hybridization to investigate the pathogenesis of aplastic anaemia. METHODS: The bone marrow mononuclear cells of a first visit aplastic anaemia patient and a healthy donor of the same age and sex were isolated using lymphocyte separating medium by density gradient centrifugation. With the patients as "tester" and donor as "driver", their CD4(+) T cells were separated with magnetic bead sorting and a cDNA library established by suppressive subtractive hybridization. Then 15 of the resulting subtracted cDNA clones were randomly selected for DNA sequencing and homological analysis. With semiquantitative RT-PCR, bone marrow samples from 20 patients with aplastic anaemia and 20 healthy donors assessed the expression levels of differentially expressed genes from SSH library. RESULTS: PCR detected 89 clones in the library containing an inserted fragment of 100 bp to 700 bp. Among 15 sequenced clones, 12 were known genes including 3 repeated genes. Compared with normal donors, there were 9/12 genes over-expressed in bone marrow CD4(+) T cells of patients with aplastic anaemia. The effects of these genes included protein synthesis, biology oxidation, signal transduction, proliferative regulation and cell migration. Not all these genes had been reported in the mechanisms of haematopoietic damage mediated by CD4(+) T cells in aplastic anaemia. CONCLUSIONS: Screening and cloning genes, which regulate functions of CD4(+) T cells, are helpful in elucidating the mechanisms of over proliferation, activation, infiltrating bone marrow and damaging haematopoietic cells of CD4(+) T cells in aplastic anaemia.

Expression of P120 catenin mRNA in Non-Hodgkin's lymphoma cell lines.

To investigate p120 catenin mRNA expression in Non-Hodgkin's lymphoma (NHL) cell lines (U937, Raji, Jurkat and Molt4) and normal lymphocytes and explore the relationship between p120 catenin and Non-Hodgkins lymphoma, total RNA sample was extracted by using TRIzol and reversely transcripted into cDNA. Polymerase chain reaction was performed to detect mRNA expression of p120 catenin in NHL cell lines U937, Raji, Jurkat and Molt4. Normal lymphocytes were used as control. It was found expressions of p120 catenin 1A and 3A mRNA were high in above-mentioned NHL cell lines, but neither p120 catenin 1A nor 3A was found in normal lymphocytes as shown by RT-PCR. It is concluded that both P120ctn1A and P120ctn3A mRNA transcripts were found in all NHL cell lines U937, Raji, Jurkat and Molt4 but they don't exist in normal lymphocytes, suggesting p120ctn possibly is of importance in diagnosis and therapy of lymphoma.

[Expression of P120ctn in non-Hodgkin's lymphoma and its significance].

To evaluate the expression of P120ctn in non-Hodgkin's lymphoma (NHL) and to explore its clinical significance, immunohistochemistry stain method was applied to comparatively investigate the protein expression of P120ctn in paraffin-embedded lymph node tissue slices from 40 cases of NHL and 10 cases of reactive hyperplasia of lymph node. The results showed that P120ctn was not detected in reactive hyperplasia of lymph node, but was detected in 55% (22/40) cases of NHL. P120ctn expression increased with the tumor malignancy of NHL, there was a significant difference between the expression rates of P120ctn in low grade (16.7%, 2/12) and intermediate to high grade malignant (71.4%, 20/28) NHL (P < 0.001). Moreover, P120ctn was also detected in vascular endothelial cells of NHL. It is concluded that the level of P120ctn expression is closely related to the malignant grade of NHL, it suggests that P120ctn possibly plays an important role in the malignant proliferation of lymphoma with a certain significance in diagnosis and therapy of lymphoma.

Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/Akt signaling pathways.

AIM: To investigate the effects of Sonic hedgehog (shh) protein on bone marrow- derived endothelial progenitor cells (BM-EPC) proliferation, migration and vascular endothelial growth factor (VEGF) production, and the potential signaling pathways involved in these effects. METHODS: Bone marrow-derived Flk-1(+) cells were enriched using the MACS system from adult Kunming mice and then BM-EPC was cultured in gelatin-coated culture dishes. The effects of shh N-terminal peptide on BM-EPC proliferation were evaluated using the MTT colorimetric assay. Cell migration was assayed using a modified Boyden chamber technique. The production of VEGF was determined by ELISA and immunofluorescence analysis. The potential involvement of PKC and PI3K signaling pathways was explored using selective inhibitor or Western blot. RESULTS: The proliferation, migration and VEGF production in BM-EPC could be promoted by endogenous shh N-terminal peptide at concentrations of 0.1 microg/mL to 10 microg/mL, and could be inhibited by anti-shh antibodies. Shh-mediated proliferation and migration in BM-EPC could be partly attenuated by anti-VEGF. Phospho-PI3-kinase expression in newly separated BM-EPC was low, and it increased significantly when exogenous shh N-terminal peptide was added, but could be attenuated by anti-human/mouse shh N-terminal peptide antibody. Moreover, the inhibitor of the PI3-kinase, but not the inhibitor of the PKC, significantly inhibited the shh-mediated proliferation, migration and VEGF production. CONCLUSION: Shh protein can stimulate bone marrow-derived BM-EPC proliferation, migration and VEGF production, which may promote neovascularization to ischemic tissues. This results also suggests that the PI3-kinase/Akt signaling pathways are involved in the angiogenic effects of shh.

[Analysis of expression of mitosis checkpoint gene chfr in bone marrow cells of acute leukemia patients].

This study was purposed to investigate the significance of mitosis checkpoint gene chfr expression in acute leukemia (AL). 2 ml of bone marrow were extracted from each of 46 AL patients and 10 normal donors as control and their mononuclear cells were isolated. Then, their chfr expression was detected by using RT-PCR and immunohistochemistry. Normal control blood samples were also analyzed. The results showed that in 15 out of 28 cases of acute non-lymphocytic leukemia and 13 out of 18 cases of acute lymphocytic leukemia expression of chfr gene mRNA and protein significantly decreased as compared with control. The cytogenetic analysis of patients with a decreased Chfr expression revealed abnormal chromosome. In conclusion, Chfr gene is a leukemia-related gene and may play an important role in leukemia pathogenesis.

Expansive effects of aorta-gonad-mesonephros-derived stromal cells on hematopoietic stem cells from embryonic stem cells.

BACKGROUND: Hematopoietic stem cells (HSCs) give rise to all blood and immune cells and are used in clinical transplantation protocols to treat a wide variety of refractory diseases, but the amplification of HSCs has been difficult to achieve in vitro. In the present study, the expansive effects of aorta-gonad-mesonephros (AGM) region derived stromal cells on HSCs were explored, attempting to improve the efficiency of HSC transplantation in clinical practice. METHODS: The murine stromal cells were isolated from the AGM region of 12 days postcoitum (dpc) murine embryos and bone marrow (BM) of 6 weeks old mice, respectively. After identification with flow cytometry and immunocytochemistry, the stromal cells were co-cultured with ESCs-derived, cytokines-induced HSCs. The maintenance and expansion of ESCs-derived HSCs were evaluated by detecting the population of CD34+ and CD34+Sca-1+ cells with flow cytometry and the blast colony-forming cells (BL-CFCs), high proliferative potential colony-forming cells (HPP-CFCs) by using semi-solid medium colonial culture. Finally, the homing and hematopoietic reconstruction abilities of HSCs were evaluated using a murine model of HSC transplantation in vivo. RESULTS: AGM and BM-derived stromal cells were morphologically and phenotypically similar, and had the features of stromal cells. When co-cultured with AGM or BM stromal cells, more primitive progenitor cells (HPP-CFCs) could be detected in ESCs derived hematopoietic precursor cells, but BL-CFC's expansion could be detected only when co-cultured with AGM-derived stromal cells. The population of CD34+ hematopoietic stem/progenitor cells were expanded 3 times, but no significant expansion in the population of CD34+Sca-1+ cells was noted when co-cultured with BM stromal cells. While both CD34+ hematopoietic stem/progenitor cells and CD34+Sca-1+ cells were expanded 4 to 5 times respectively when co-cultured with AGM stromal cells. AGM region-derived stromal cells, like BM-derived stromal cells, could promote hematopoietic reconstruction and HSCs' homing to BM in vivo. CONCLUSIONS: AGM-derived stromal cells in comparison with the BM-derived stromal cells could not only support the expansion of HSCs but also maintain the self-renewal and multi-lineage differentiation more effectively. They are promising in HSC transplantation.

Methylation of gene CHFR promoter in acute leukemia cells.

In order to explore whether gene CHFR was inactivated by methylation in leukemia cells, the expression of CHFR was examined before and after treatment with demethylation agent in Molt-4, Jurkat and U937 leukemia cell lines by means of RT-PCR. The methylation of promoter in Molt-4, Jurkat and U937 cells as well as 41 acute leukemia patients was analyzed by MS-PCR. The results showed that methylation of CHFR promoter was inactivated and could be reversed by treatment with a demethylating agent in Molt-4, Jurkat and U937. CHFR promoter methylation was detected in 39% of acute leukemia patients. There was no difference in incidence of CHFR promoter methylation between acute myelocytic leukemia and acute lymphocytic leukemia. In conclusion, CHFR is frequently inactivated in acute leukemia and is a good candidate for the leukemia supper gene. By affecting mitotic checkpoint function, CHFR inactivation likely plays a key role in tumorigenesis in acute leukemia. Moreover, the methylation of gene CHFR appears to be a good index with which to predict the sensitivity of acute leukemia to microtubule inhibitors.

[Expression of mitosis checkpoint gene CHFR in acute leukemia].

OBJECTIVE: To investigate the expression of mitosis checkpoint gene CHFR in adult patients with acute leukemia (AL) and its clinical significance. METHODS: Four ml of bone marrow was extracted from 65 AL patients, 38 males and 27 females, with the median age of 35, 43 with acute myelocytic leukemia (AML) and 22 with acute lymphocytic leukemia (ALL), 45 de novo patients and 20 recurrent patients, and 8 normal donor of allogeneic bone marrow transplantation as controls. The bone marrow mononuclear cells (BMNC) were isolated. The cell cycle was examined by flow cytometric analysis. The CHFR mRNA level in the BMNC was measured by RT-PCR. The expression of CHFR protein was detected by Western blotting in 32 of the 65 patients and 8 normal persons as control. RESULTS: (1) The levels of CHFR protein and mRNA were correlated with the cumulative percentages of cells in S phases. (2) The expression level of CHFR protein in 40.6% (13/32) of the AL patients and that of the CHFR mRNA in 60.0% (27/45) of the AL patients were both significantly lower than those of the normal controls. (3) The mean expression level of CHFR protein in the recurrent acute lymphoblastic leukemia (ALL) was 0.71, significantly higher than that of the de novo group (0.38, t = 2.54, P = 0.017). (4) The complete remission (CR) rates in the AL patients with high expression levels of CHFR protein and mRNA were 30.2% and 42.4% respectively, significantly lower than those in the AL patients with low expression levels (88.6% and 85.4% respectively, both P < 0.05). Multivariate analysis showed that CHFR was one of the influencing factors of CR rate of AL patients. CONCLUSION: By affecting mitotic checkpoint function, CHFR inactivation plays a key role in tumorigenesis in adult patients with acute leukemia. Moreover, the aberrant expression of CHFR appears to be a good molecular marker to predict the sensitivity of acute leukemia to chemotherapy.

Transfection efficiency comparison of oligonucleotide and plasmid to the HL-60 cell line with liposomes.

The transfection efficiency of oligonucleotide and plasmid to the HL-60 cell line with lipofectaminePLUS was compared through observing the transfection rate and the expression duration of exogenous gene in the target cells. The results showed that the transfection rate of oligonucleotide to the HL-60 was about 90% - 95% and it had no obvious attenuation within 84 h. However, the plasmid transfection rate was only 5% -25% and it was decreased significantly within 60 h. It was suggested that the transfection of oligonucleotide with liposomes was better than that of plasmid.

[Effects of ligustrazine on expression of VCAM-1/VLA-4 in syngenic bone marrow transplantation of mice].

To explore the effect of ligustrazine on the expression of adherent molecule VCAM-1/VLA-4 of bone marrow cells in syngenic bone marrow transplantation (BMT) mice, the mice were divided into 3 groups: normal group (which received no treatment), BMT control group and ligustrazine-treated groups. BMT mouse models were established. The BMT control group and the ligustrazine-treated group were orally administered 0.2 ml saline per mouse and 2 mg ligustrazine per mouse, respectively, twice a day. On the day 7, 14, 21, 28 after BMT, mice were respectively killed. Bone marrow nucleated cells were detected, and then the expression of VCAM-1/VLA-4 was assayed by immunohistochemistry, RT-PCR and flow cytometry analysis, respectively. The results showed that in ligustrazine-treated group, the accounts of bone marrow nucleated cells on the day 7, 14, 21, 28 after BMT were all higher than that in BMT control group. The expression level in the ligustrazine-treated group was significantly higher than that in the BMT control group (P < 0.05 or P < 0.01). It is concluded that ligustrazine can enhance VCAM-1/VLA-4 expression in bone marrow after syngenic bone marrow transplantation in mice, which may be related to the mechanisms underlying the ligustrazine accelerating hematopoietic reconstitution in allogenic bone marrow transplantation.

Expression of CD28 and CTLA4 on T cells in bone morrow of immune-mediated aplastic anemia mice.

To investigate the expression and significance of CD28 and CTLA4 on T cells in bone marrow of aplastic anemia (AA) mice, in vitro bone marrow mononuclear cells (BMMNCs) were activated through being incubated with PHA (15 microg/mL). The expression of CD28 and CTLA4 on T cells incubated with or without PHA was detected by two-color flow cytometry. The expression of CD28 and CTLA4 was significantly increased after PHA stimulation. In the AA mice, the expression of CD28 with or without PHA stimulation was both higher than that in the normal mice (both P < 0.01), but the expression of CTLA4 with or without PHA stimulation showed no significant difference in comparison to that in the normal mice (both P > 0.05). In the AA mice, there were more activation and activated potential of T cells than the normal, and the abnormal expression of CD28 and CTLA4 may participate in immunological disorder mediated by T cells.