Objective To compare the effect of perineal cleansing with the potassium permanganate or sterile water on mid- stream urine culture. Methods Mid- stream specimens of urine were obtained from inpatients in our hospital between January 2002 and December 2006. All these patients may be diag-nosed as urinary tract infection. The urine specimens were divided into the potassium permanganate group (n=1572, the sterilization group) and the sterile water group (n=544). The change of positive and contami-nation rate of mid-stream urine culture from the specimens was observed. More than two kinds of germs in one urine specimen were defined as contamination. Results 830 patients with urinary tract infection had been enrolled. 2116 specimens were collected and 531 strains of causative organism were detected. The positive rate of the sterilization group and the sterile water group was 20.04% and 39.71%, respectively,and such difference was significant. The rate of identical causative organism from the same patient whose spec-imen was cultivated twice in the sterilization group was 0.012% and the rate was 0.105% in the sterile water group. The difference was significant. The rate of different or one kind of causative organism from the same patient whose specimen was cultivated twice in these two groups hadn't significant deviation. The contami-nation rate of the sterilization group (0.028%) was significantly higher than that of the sterile water group (0.007%). Conclusions Perineal cleansing with sterile water can reduce the false negative rate of mid-stream urine culture without increasing the contamination rate. Potassium permanganate sterilization is re-sponsible for the high false-negative in mid-stream urine culture.

Human Interleukin-11(hIL-11)has no Cys residue in its natural form.By site-directed mutagenesis,a Cys residue can be introduced to replace the 1st residue Gly and the rhIL-11 was chemically modified by using 20 kDa mPEG-maleimide conjugated to this site.The mPEG-hIL-11 conjugate was purified and showed a single band on SDS-PAGE with an apparent molecular weight.The biological activity of purified mPEG-hIL-11 was determined using a dependent cell line 7TD1.The remaining biological activity of PEGylated-rhIL-11 was 30% of native rhIL-11,suggesting chemical modification of rhIL-11 by PEG is a promising approach for improving the pharmacological efficacy.

This study was aimed to investigate the effect of cord blood dendritic cells (DCs) on the in vitro proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia cells of the homologous cytokine-induced killer (CIK) cells. DCs and CIK cells were induced from cord blood mononuclear cells. They were co-cultured at the ratio of 1:5, and CIK cells from cord blood or DC-CIK cells from peripheral blood were cultured as controls. Immunophenotypic changes were analyzed by flow cytometry, increased number of cells were counted by trypan-blue staining, the killing activity to leukemia cells was assayed by MTT, the levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-12 (IL-12) in the cultured supernatant were detected by ELISA. The results showed that the proliferation capability of cord blood DC-CIK cells was significantly higher than that of cord blood CIK cells and peripheral blood DC-CIK cells (p < 0.05 and p < 0.05). Under the same condition, the rate of double positive cells with CD3(+)CD8(+) and CD3(+)CD56(+) in CIK cells was significantly enhanced by co-culture with cord blood DCs (p < 0.05). The level of IL-12, IFN-γ, and TNF-α in cultured supernatants of cord blood DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone (p < 0.01, p < 0.05, p < 0.05). Within the effector-target ratio range between 2.5:1 to 20:1, the activity of cord blood DC-CIK cells against all subtypes of acute leukemia cells was much higher than that of CIK cells (p < 0.05), and there was no significant difference among all subtypes of acute leukemia cells, which was the same with the killing effect of peripheral blood DC-CIK cells against leukemia cells. It is concluded that the proliferation capability and anti-leukemia effect of the homologous CIK cells can be enhanced by cord blood DCs. The proliferation capability of cord blood DC-CIK cells is stronger than that of peripheral blood DC-CIK cells, but there is no significant differences of cytotoxicity between DCs and CIK cells. As the cord blood is easily gained and does not easily cause a serious graft rejection, the DC-CIK cells should be clinically applied more extensively as novel immune therapy.
Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Leukemia ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

This study was aimed to investigate the effect of dendritic cells (DC) on the proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia of cytokine-induced killer (CIK) cells in vitro. DCs and CIK cells were induced from peripheral blood mononuclear cells of healthy volunteers. They were co-cultured meanwhile CIK cells were cultured alone as controls. Increased number of cells were counted by trypan-blue staining; the killing activity was detected by MTT assay; immunophenotype changes were analyzed by flow cytometry; the IL-12 and INF-gamma levels of the cultured supernatants were detected by ELISA kits. The results showed that the proliferation capability of DC-CIK cells was significantly higher than that of CIK cells (p < 0.05). Under the same condition, the ratio of double positive cells such as CD3(+) CD8(+), CD3(+) CD56(+) in CIK cells was significantly enhanced by co-cultured with DC cells (p < 0.05). The levels of IL-12 and INF-gamma in cultured supernatants of DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone (p < 0.01, p < 0.05). Within the effector-target ratio range between 5:1 to 40:1, the activity of DC-CIK cells against leukemia cells were much higher than that of CIK cells (p < 0.05), and this effect showed a positive correlation with the effector-target ratio. It is concluded that the proliferation capability of DC-CIK cells, the level of their secreted cytokines and their activity against leukemia cells are significantly higher than those of CIK cells. This research may suggest an approach for clinical immunotherapy against leukemia with DC-CIK cells.
Cell Line, Tumor ; Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; metabolism ; Dendritic Cells ; cytology ; immunology ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism

To study the influence of IFN-alpha on function of CML-DC cultured in vitro and expression of chemokine and its chemokine receptor, bone marrow mononuclear cells from 13 CML patients were cultured in the fetal calf serum culture system supplemented with rhSCF, rhFlt-3L for expansion system, and adding rhGM-CSF, rhTNF-alpha, rhIL-4, with or without rhIFN-alpha to induce DCs. After incubation for two weeks, the phenotypes of CML-DC were analyzed by direct immunofluorescence and flow cytometry. The concentration of MIP-3beta expressed by CML-DC in the supernatant were analyzed by ELISA. The proliferative ability of T cells from healthy volunteers stimulated by CML-DCs were measured by MTT assay. The results showed that expression of CD86, CD83, CD40, MHC-I class molecules, CCR7, the concentration of MIP-3beta expressed by CML-DC, and the proliferative ability of T cells stimulated by CML-DCs in IFN-alpha group were all significantly higher than that in control group (P < 0.01). It is concluded that the immunophenotype of CML-DCs can be partially changed by IFN-alpha to accelerate the maturation of CML-DCs, enhance the capacity of CML-DCs, and stimulate allogeneic T lymphocyte proliferation.
Adult ; Aged ; Antigens, CD ; analysis ; B7-2 Antigen ; analysis ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; CD40 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chemokines ; biosynthesis ; Dendritic Cells ; drug effects ; metabolism ; pathology ; Female ; Flow Cytometry ; Fluorescent Antibody Technique, Direct ; Humans ; Immunoglobulins ; analysis ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; Leukocytes, Mononuclear ; drug effects ; metabolism ; pathology ; Male ; Membrane Glycoproteins ; analysis ; Middle Aged ; Receptors, Chemokine ; biosynthesis

The study was aimed to investigate the influence of interferon alpha (IFN-alpha) on the expressions of Fas and Fas ligand (FasL) in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to adding stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF-alpha) and interleukin 4 (IL-4), the IFN-alpha was added to the serum-free medium for DCs. After culturing for 10 - 14 days, cell phenotype and percentage of Ph(1) chromosome were detected by different methods. The expression of Fas or FasL on CML-DCs and cell cycle of DCs labeled with propidium iodine (PI) were measured by flow cytometry. The concentration of sFas in supernatants was analyzed by enzyme-linked immunosorbent assay (ELISA). The results indicated that the expression of co-stimulatory molecules were improved significantly while the percentages of Ph(1) positive cells decreased. The level of Fas on cells was up-regulated and the concentration of sFas decreased. However, the expression of FasL was negative. The ratio of apoptosis rose gradually while the concentration of IFN-alpha increased. It is concluded that IFN-alpha can accelerate the apoptosis of Ph(1) positive cells through Fas/FasL pathway, so the number of Ph(1) negative cells increases relatively.
Adolescent ; Adult ; Apoptosis ; drug effects ; Cells, Cultured ; Child ; Culture Media ; pharmacology ; Dendritic Cells ; cytology ; metabolism ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Humans ; Interferon-alpha ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Middle Aged ; Philadelphia Chromosome ; Young Adult ; fas Receptor ; genetics ; metabolism

This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN-alpha. The mechanism may lie in the facts that expression of CCR7 and co-stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha.
Cells, Cultured ; Dendritic Cells ; cytology ; Humans ; Interferon-alpha ; pharmacology ; Interleukin-10 ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; Philadelphia Chromosome ; drug effects ; Receptors, CCR7 ; genetics ; metabolism

The study was aimed to investigate the extensive amplification and the cytotoxicity of dendritic cells (DC) derived from chronic myeloid leukemia cells. DC were cultured in two steps: firstly, extensive amplification in primary culture of CD34(+) or mononuclear cells isolated from CML patients' bone marrow and peripheral blood with rhFlt3-L and rhTPO for 7 days; secondly, inducing culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. A system inducing DC directly were established for comparison. DC were identified by immunophenotype with flow cytometry, chromosome analysis by displaying G banding and electric microscopy analysis. The function of stimulating T cells proliferation and cytotoxicity of CML cells were confirmed through MTT assay. The results showed that after first extensive amplification in primary culture with rhFlt3-L and rhTPO for 7 days, CD34(+) cells had a total cell number with (77 +/- 5) fold expansion, and DC were (39 +/- 8)% of total cell respectively after induction culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. Both the amplification of cell number and yield of DC were higher than the system without extensively culture (P < 0.01). Such DC could stimulate T cells to proliferate and kill leukemia cells finally. In conclusion, two-step culture method can obviously improve the cell number of DC required, that is better than inducing them directly. DC derived from CML cells induce the generation of anti-leukemia immunization.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; immunology ; Cell Proliferation ; Cells, Cultured ; Child ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; pathology ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; pathology ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the expression and its clinical significance of estrogen receptor (ERα) and phosphorylated estrogen receptor (p-ERα) in patients with hepatocellular carcinoma. The associations between ERα, p-ERα and IL-6 were also analyzed.

METHODSImmunohistochemistry was used to detect the expression of ERα, p-ERα and IL-6 in tumor tissues from 77 cases with hepatocellular carcinoma. The relations between ERα and the clinical pathological parameters and prognosis were also analyzed.

RESULTSThe positive rates of ERα, p-ERα and IL-6 in hepatocellular carcinoma were 39.0% (30/77), 45.4% (35/77) and 72.7% (56/77), respectively. The expression of ERα and p-ERα were negatively correlated with the expression of IL-6 (r=-0.468, P<0.01; r=-0.370, P<0.01, respectively). The positive rate of ERα in patients with tumor size≤5 cm, serum level of alpha-fetoprotein<400 µg/L, with complete encapsulation and non-microvascular invasion was significantly higher than those with tumor size>5 cm, serum level of alpha-fetoprotein≥400 µg/L, non-complete encapsulation and with microvascular invasion (all P<0.05). The overall survival rates of ERα-positive and ERα-negative patients were 66.7% and 23.4% (P<0.05). And the disease-free survival rates of ERα-positive and ERα-negative patients were 83.3% and 57.4% (P<0.05).

CONCLUSIONSThe tumor biological features of ERα-positive patients are better than that of ERα-negative patients. The role of ERα in hepatocellular carcinoma may be related to IL-6 level.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Estrogen Receptor alpha ; metabolism ; Female ; Hepatitis B ; metabolism ; pathology ; Humans ; Interleukin-6 ; metabolism ; Kaplan-Meier Estimate ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Phosphorylation ; Prognosis ; Proportional Hazards Models ; Young Adult

Objective　To observe the characteristics and rules of craniocerebral injury resulting from a high explosive shell to provide the bases for treating explosive injury. Methods　A total of 36 sheep were distributed at the distance 6 to 48 m away from the explosive center and the shell was exploded electrically at 7 m above the earth. At the same time, the velocity of fragments and shock wave pressure were determined. Gross and pathological observations were performed after injury. Results　Among all sheep with fragment injury, craniocerebral injury was 32%. Their immediate death rate was 75% and all died 6 h later. The incidence rates of penetrating wound and blind wound were 75% and 25% respectively. Pollution of wound track was heavy. The percentage of head lost was 50% in sheep and 50% of injured animal suffered from comminuted fracture of skull base. Bleeding was found extensively on the surface of the cerebrum, even medulla oblongata was involved. Hemorrhage, edema, rupture of small blood vessels and degeneration of neuron were found at the regions 4 cm away from the wound tract with light microscopy. Combined blast injury was found and occurred most often in the abdomen and limbs, both accounting for 62.5%, and combined thoracic injury was the third, up to 50%. All the animals of craniocerebral injury combined with lung blast injury. Conclusion　High explosive shells destroy cranium badly and extensively. Many skulls are lost and the cranial base is readily fractured. The wound track is heavily polluted. Combined injury is more often occurred.