Objective To explore the clinical value of serum procalcitonin (PCT)-based antibiotic therapy in the second-classedexacerbations of chronic obstructive pulmonary disease (AECOPD). Methods 240 patients diagnosised as AECOPD were randomized to the PCT group and the control group. Serum PCT levels of patients from the PCT group were measured 1 h after hospitalized and the third, fifth, eighth day respectively. When PCT < 0.1 μg / L, patients will stop taking antibiotics and initiated while PCT≥0.1 μg / L. Antibiotic treatment in the control group was based on guidelines of COPD diagnosis and treatment. Results Duration of antibiotic therapy and hospitalization were respectively 5.6 ± 1.4 and 8.2 ± 1.1 days in the PCT group, 9.2 ± 2.2 and 11.4 ± 2.5 days in the control group (both P < 0.05). Mean costs of hospitalization expensesand antibiotic therapy were 5700 ± 201 and 1650 ± 189) yuan in the PCT group, 6210 ± 220 and 2350 ± 210 yuan in the control group (both P < 0.05). The clinical effective rate, times of exacerbation, one-year ΔFEV1, the 1-year hospitalization rate and time to next exacerbation all showed no significant differences between the two groups. Conclusion PCT-guided antibiotic treatment reduces antibiotic use inthe second-classed acute exacerbations patients.

OBJECTIVETo study, the high polar media-water extraction process of Herba epimedii with the microwave technology.

METHODThe single parameters of extraction process was investigated with dry extract yield and extraction efficiency of icariin as content index.

RESULTThrough studying the single parameter, the optimal conditions are as follows: 70% ethanol as wetting solvent, volume of solvent to weight of material equal to 6:1 in wetting, 30 min wetting time, 2.5 min microwave irradiation time, volume of solvent to weight of material equal to 50:1 in extraction process, 10 min microwave-assisted extraction time.

CONCLUSIONCompared with conventional methods, microwave-assisted extraction (MAE) after Microwave Pretreament (MP) of raw material was higher extraction efficiency and time-saving.

Drugs, Chinese Herbal ; analysis ; isolation & purification ; Epimedium ; chemistry ; Flavonoids ; analysis ; isolation & purification ; Microwaves ; Plants, Medicinal ; chemistry ; Solvents ; Technology, Pharmaceutical ; methods

OBJECTIVETo study changes to CD4(+)CD25(high+)CD127(low) regulatory T cells (Treg) in peripheral blood from children with bronchiolitis, and to explore its clinical significance.

METHODSThirty-one children with bronchiolitis and aged under two years were randomly enrolled as the bronchiolitis group, and 25 under two-year-olds with bronchopneumonia were randomly enrolled as the bronchopneumonia group. A further twenty-five children with non-infectious diseases such as hernia and renal calculus served as the control group. The level of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood was measured by multi-color detection and multi-parameter flow cytometry.

RESULTSThe proportion of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood in the bronchiolitis group (8.0%±2.1%) was significantly lower than in the bronchopneumonia (9.6%±2.6%; P<0.05) and control groups (11.3%±2.9%; P<0.05).

CONCLUSIONSCD4(+)CD25(high+)CD127(low) Treg level in peripheral blood may be an index of immunological function in infants. A decreased level of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood suggests that Treg cells may be involved in the pathogenesis and development of bronchiolitis.

Bronchiolitis ; immunology ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; blood ; Interleukin-7 Receptor alpha Subunit ; blood ; Male ; T-Lymphocytes, Regulatory ; immunology

Aged ; Coal Mining ; Drug Combinations ; Humans ; Male ; Middle Aged ; Peroxides ; administration & dosage ; therapeutic use ; Pneumoconiosis ; complications ; drug therapy ; Respiratory Insufficiency ; drug therapy ; etiology ; Urea ; administration & dosage ; analogs & derivatives ; therapeutic use

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

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

To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.
Animals ; Antiviral Agents ; metabolism ; Baculoviridae ; genetics ; Cell Line ; Cloning, Molecular ; Gene Expression ; Gene Fusion ; Genetic Vectors ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Insecta ; Interferon-alpha ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the role of multidrug resistant protein 2 (MRP2) and glutathione (GSH) cotransport system in hepatic arsenic metabolism in rats.

METHODSThirty healthy Wistar rats were divided randomizedly into five groups. The first group was the control group and the rats in this group were administered with normal saline. In the second, third and fourth group the rats were administered with 4, 10 and 20 mg As(+)3/kg BW of sodium arsenite respectively every other day for two weeks. The fifth group was the benzene-soluble organics (BSO) intervention group and in this group the rats were administered with 2 mmol/kg BW BSO intraperitoneally every day three days before the end of the experiment. The other treatment was the same as in other groups. All rats were sacrificed two weeks after the treatments. Arsenic contents in bile, liver and blood were detected by atomic absorption spectroscopy (AAS), and the expression of MRP2 in the membrane of hepatocyte was determined by Western-blot analysis.

RESULTSThe level of total arsenic (including organic arsenic and inorganic arsenic) in bile, liver and blood in all three different dose groups was higher than those in the control groups (P < 0.05). Arsenic levels of bile and liver were increased with intragastric arsenic dose. Blood arsenic levels were not significantly different in three different dose groups. Expression of hepatic MRP2 was increased with intragastric arsenic concentration. A positive correlation between biliary arsenic concentration and MRP2 levels was found in liver (r = 0.986, P < 0.05). For the rats pretreated with BSO, the biliary arsenic was significantly higher than that in the control group but lower than that in the high dose group; the liver and blood arsenic was higher than that in the control group and in the high dose group. Expression of MRP2 pretreated with BSO was decreased.

CONCLUSIONSodium arsenite can induce expression of MRP2 and the up-regulation of MRP2 may play an important role in the bile secretion of arsenite and its metabolites. The function of MRP2 for transportation of arsenic and its metabolites is associated with the intracellular GSH level. BSO inhibits the synthesis of GSH, which weakens the function of the MRP2-GSH cotransport system and makes the liver arsenic increased.

Animals ; Arsenic ; pharmacokinetics ; Arsenic Poisoning ; metabolism ; Bile ; metabolism ; Female ; Glutathione ; biosynthesis ; Liver ; metabolism ; Male ; Membrane Transport Proteins ; biosynthesis ; Multidrug Resistance-Associated Proteins ; biosynthesis ; Random Allocation ; Rats ; Up-Regulation

Objective To explore the clinical significance of detecting serum interleukin-10(IL-10) level and the proportion of myeloid-derived suppressor cells (MDSCs) in peripheral blood mononuclear cells (PBMCs) in children with bronchiolitis.Methods Fifty-one children with bronchiolitis less than 2 years old were randomly enrolled including 27 boys and 24 girls.They were divided into 2 groups:bronchiolitis group Ⅰ,25 children with atopic high risks were included in this group;bronchiolitis group Ⅱ,26 children without atopic high risks were included in this group.Children without infectious diseases such as hernia and renal calculus had been randomly enrolled as the control group (without atopic disease and atopic family disease) (n =45),including 25 boys and 20 girls.After taking 4 mL venous blood of patients in 3 groups,1 mL was used to test the serum proportion of MDSCs by flow cytometry,and the remaining blood was used to test the level of IL-10 in the serum by enzyme-linked immunosorbent assay.Results 1.The proportions of MDSCs in the PBMCs in bronchiolitis group Ⅰ [(3.17 ± 0.24) ％] and bronchiolitis group Ⅱ [(1.33 ±0.25) ％] were significantly higher than that of control group [(0.78 ± 0.25) ％] (all P ＜ 0.01),and the proportion of MDSCs in the PBMCs in bronchiolitis group Ⅰ was higher than that of bronchiolitis group Ⅱ (P ＜0.01).2.The serum levels of IL-10 in bronchiolitis bronchiolitis group Ⅰ [(31.88-± 3.91) ng/L] and bronchiolitis group Ⅱ [(23.85 ±4.10) ng/L] were significantly higher than that of control group [(13.63 ± 2.83) ng/L] (all P ＜0.01),and the levels of IL-10 in bronchiolitis group Ⅰ was higher than that of bronchiolitis group Ⅱ (P ＜ 0.01).3.There was a positive correlation between the proportion of MDSCs in the PBMCs and the serum levels of IL-10 in bronchiolitis group Ⅰ (r =0.717,P ＜ 0.01),but there was not any correlation between bronchiolitis group Ⅱ and control group (r =0.262,-0.102,all P ＞ 0.05).Conclusion MDSCs plays a crucial role by up-regulating the IL-10 level in the process of developing asthma from bronchiolitis.

BACKGROUNDTrichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dermatophytosis caused by T. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin ®, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection.

METHODSThe reconstructed human epidermis (RHE) was infected by inoculating low-dose (400 conidia) and high-dose (4000 conidia) T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy (SEM) observation.

RESULTSThe histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of T. rubrum infection in an intuitionistic way.

CONCLUSIONSWe established the T. rubrum infection model on RHE in vitro successfully. It is a promising model for further investigation of the mechanisms involved in T. rubrum infection.

Animals ; Disease Models, Animal ; Epidermis ; microbiology ; Humans ; Keratinocytes ; cytology ; Tissue Culture Techniques ; Trichophyton ; pathogenicity