Objective To explore the quality of life and its related factors of patients with asthma. Methods 60 asthma patients were assessed by Hamilton Rating Scale for Depression(HAMD),Hamilton Rating Scale for Anxiety(HAMA)and the Short-Form-36 Health Survey(SF-36). Results Except physical functioning (PF) domain, the SF-36 score of asthma patients was significantly lower than that of normal. The illness course longer than 4 years had lower score, so did the quality of life. The SF-36 score of asthma patients with depression symptom and anxiety symptom was lower than those without depression symptom and anxiety symptom. Conclusions The asthma patients' quality of life are worse than those of normal, especially the patients with long illness course, depression symptom and anxiety symptom.

Objective:To evaluate the in vitro and in vivo immunostimulatory activity and the safety of ethanol extract of wild Artemisia rupestris L. (EEWAR). Methods:Bone marrow dendritic cells (BMDCs) from C57BL/6 mice were treated with different concentrations of EEWAR in vitro and the expression of CD40 and CD80 on BMDCs was detected by flow cytometry. ICR mice were subcutaneously immunized with different concentrations of EEWAR in combination with ovalbumin (OVA) or OVA alone. Aluminum adjuvant was used as the positive control. OVA-specific IgG antibodies in mouse serum samples were measured by ELISA following immunization. T cell proliferation in spleen tissues was detected by MTT method. Acute toxicity test was conducted in ICR mice to analyze the safety of EEWAR. Results:In vitro experiment showed that EEWAR at the concentrations of 10-20 μg/ml increased the expression of CD40 and CD80 on BMDCs ( P<0.05), and had no significant effect on the morphology of BMDCs; EEWAR at the concentrations of 100-200 μg/ml significantly promoted the expression of CD40 and CD80 on BMDCs ( P<0.01), but had a certain influence on the morphology of BMDCs. In vivo experiment showed that EEWAR enhanced the production of IgG, IgG 1 and IgG 2a antibodies against OVA and the proliferation of splenocytes ( P<0.05). In the acute toxicity test, EEWAR at the concentrations of 50-5 000 μg/ml had no side effects on mouse body weight and was relatively safe. Conclusions:EEWAR could promote the maturation of DCs and enhance the humoral and cellular immune responses when used as an adjuvant to OVA. It was safe in a certain dose range. This study provided reference for further research on EEWAR as a new-generation adjuvant.

Objective To investigate the influence of salmeterol with fluticasone on airway inflammation of patients with acute exacerbation chronic obstructive pulmonary disease (COPD).Methods 40 COPD patients in the stage of acute exacerbation were randomly divided into salmeterol with fluticasone group( A group) and controll group (B group).Bronchial alveolar lavage( BAL) was performed as usual.The concentrations of IL-8 and TNF-α in bronchial alveolar lavage fluid(BALF) were measured by ELISA.The results were compared with that of 18 healthy volunteers.Results The levels of IL-8 and TNF-a in BALF of patients in A group(10.60 ± 1.42) μg/L, (14.80 ± 2.05) μg/Land B group( 10.77 ± 1.98) μg/L, (14.70 ± 2.03) μg/L were significantly higher than that of C group (3.40 ±0.65)μg/L, (4.67 ± 1.01) μg/L( all P <0.01) ;Before treatment,the levels of IL-8 and INF-α in BALF of A group( 10.60 ± 1.42)μg/L,(14.80 ±2.05) μg/L and B group( 10.77 ± 1.98) μg/L,(14.70 ±2.03) μg/L had no significant differences (all P > 0.05) ,and the concentrations of IL-8 and TNF-a in BALF in group A (4.39 ± 0.92)μg/L,(5.84 ±1.26) μg/L were significantly lower than that in group B(9.69 ± 1.43) μg/L, (12.88 ± 2.51) μg/L after two weeks treatment ( all P < 0.01).Conclusion Salmeterol with fluticasone could inhibit airway inflammation of COPD patients in the stage of acute exacerbation.

Purpose To identify whether serum-free culture can enrich breast cancer stem cells from MCF-7 human breast cancer cell line. Methods MCF-7 human breast cancer cell line by serum-free culture and serum culture technology were cultured, its cell mor-phology and growth pattern were observed by the inverted microscope. The expression of stem cell surface molecular makers CD24, CD44 was observed by the inverted fluorescence microscope and the flow cytometry, the proportion of different subpopulation cells was detected by the flow cytometry. At the same time, difference of the cell cycle was detected by the flow cytometry. Results The cell line cultured by serum-free culture grew in the form of suspended microspheres of different sizes in the medium, but the cell line cul-tured by serum culture grew in the form of monolayer adherent growth. There was no obvious difference in the expression of stem cell surface molecular makers CD44 between the suspended microsphere cells and the adherent cells, but the expression of CD24 in the sus-pended microsphere cells decreased compared to the adherent cells. The proportion of CD44 + /CD24 -/low phenotype cells in the suspen-ded microsphere cells and the adherent cells was (86. 93 ± 0. 53)% and (19. 98 ± 0. 62)%, respectively (P<0. 05), the proportion of CD44 + /CD24 + phenotype cells was (12. 68 ± 0. 59)% and (79. 90 ± 0. 57)%, respectively (P<0. 05). The proportion of mitotic cells in the suspended microsphere cells and the adherent cells was ( 18. 85 ± 2. 26 )% and ( 43. 91 ± 1. 81 )%, respectively ( P<0. 05), the proportion of quiescent cells was (64. 92 ± 2. 07)% and (39. 82 ± 1. 77)%, respectively (P<0. 05). Conclusion CD44 + /CD24 -/low breast cancer stem cells can be effectively enriched by the serum-free culture technology.

Objective To study the influence of seminal plasma reactive oxygen species(ROS),cell factors and sperm quality in infertile male with Uu infection and explore its action mechanism in male infertility.Methods Chose 83 cases of male in-fertility with Uu infection as the experimental group (Uu+ infertility group),30 cases of male infertility without Uu infec-tion (Uu- infertility group)and 30 normal men with children as a control (Normal fertility group).Respectively,determi-nate the levels of seminal plasma malondialdehyde (MDA),superoxide dismutase (SOD),IL-6,IL-10,IL-18 and TNF-α,and analyzed its correlation.Results In Uu+ infertility group,the levels of MDA (19.56±5.22 nmol/ml),IL-6 (58.31±8.94 pg/ml),IL-18 (38.16±17.02 pg/ml)and TNF-α(42.68±11.18 pg/ml)were obviously higher than those in the other two groups (t=4.35~20.43,P value<0.001),and the level of IL-10 (8.62±2.98 pg/ml)and SOD (95.36±20.03 μmol/L) was lower than those in the other two groups (t=3.67~23.74,P value<0.001).Correlation analysis found that MDA of Uu+ infertility group was positively correlated with TNF-α,IL-18 (r=0.61,0.55,P value<0.001),and negatively correla-ted with SOD and IL-10 (r=-0.55,-0.53,P value<0.001).Conclusion The results suggested that Uu infection caused the level of reactive oxidative increasing,cytokines counterbalance disorder and affected sperm quality.So it is of great signif-icance for the treatment to test the levels of ROS and cytokines in patients with male sterility.

BACKGROUND: Recently, one focus of research has been Annexin Ⅴ (AnV) existing on hepatic cells membranes as a fundamental receptor related to hepatitis B virus (HBV) infection. Also its expression in placental tissues has been a matter of debate. The study of the relationships between placental cells infected with HBV and their AnV expression will be of great value in future prevention strategies and treatments.OBJECTIVE: To investigate the presence of AnV in HBV infected human's placental cells and its potential role in HBV intrauterine transmission.DESIGN: Randomized controlled experiment.SETTING: Taishan Medical College.MATERIALS: Placental tissue was collected from HBsAg positive full term pregnant women (30 cases) admitted to Jinan Institute for Maternal and Child Health, Taian Central Hospital and Taian Institute for Maternal and Child Health from January 2003 to December 2004. Maternal serum was also obtained. Informed consents for participating in this study were obtained from all the involved pregnant women and this experiment was approved by the Hospital Ethics Committee. Rabbit-anti-human AnV purified affinity antibody (first antibody), rat-anti-human HBs mAb (first antibody),and biotinylated goat-anti-mouse IgG (secondary antibody) were supplied by Wuhan Boster Bioengineering Company.METHODS: Using SABC immunohistochemical staining reagent, 18 HBsAg positive placentas were obtained from 30HBsAg infected patients in full term pregnancy. These were considered as the positive group and the other 12 were used as negative controls. The staining process included dewaxing, dehydration of embedded slides and microwave antigen restoration. In the wet box, rabbit-anti-human AnV purified antibody (first antibody, 1:60, monoclonal antibody)was added on the slides and kept at 4 ℃ overnight. Rat-anti-human antibody HBs mAb(secondary antibody, 1:50) was added and kept at 4 ℃ ovemight, after this procedure, biotinylated goat-anti-mouse IgG(1:100), the first fluorescent antibody such as FITC-goat anti-rabbit IgG (1:50) and the second fluorescent antibody (Avidin-Cy3) were used,respectively. The slides were sealed with buffered glycerol and examined under a confocal laser scanning microscope.The images on the slides were analyzed with IPP 4.5 image programs.MAIN OUTCOME MEASURES: Detecting the simultaneous existence and distribution of HBsAg/AnV in placental cells with HBV infection.RESULTS: Ten cases from the positive group were simultaneously detected for HBsAg/AnV by double-labeled immunofluorenscence assay and confocal laser scanning microscope. AnV expression was detected in the trophoblastic, interstitial cells and vascular endothelial cells of villi interstitial blood vessels, and the coexistence of HBsAg/AnV was found even in one cell.CONCLUSION: HBsAg combined with the receptor AnV in the same placental cells is a common finding in HBV infected full term pregnant women. This finding is very suggestive of a mechanism where AnV could promote hepatitis B virus to enter the placental cells and cause intrauterine infection.

Objective To investigate the efficacy of using Xinjiang wild Artemisia rupestris L. crude polysaccharides ( WARCP) as an immunologic adjuvant for influenza virus vaccine( IVV) .Methods ICR mice were subcutaneously immunized with 0.3 μg of IVV and 1.5 μg of IVV alone or co-administered with 200 μg of WARCP on 0 d and 14 d.Antibody levels in serum samples were detected by using indirect ELISA.MTT method was used to measure the proliferation of splenocytes.The growth conditions of mice were observed as well.Results No significant differences in the body weight were observed between mice from different groups (P>0.05).The levels of influenza virus-specific IgG, IgG1 and IgG2a were signifi-cantly increased in mice injected with WARCP adjuvant (P<0.05).The levels of IgG antibody in mice im-munized with low-dose of IVV and WARCP were significantly higher than those in mice immunized with high-dose of IVV alone (P<0.05), indicating at least 80% reduction in vaccine dosage by adding WARCP as adjuvant.Moreover, WARCP significantly promoted the proliferation of lymphocytes (P<0.05).Conclu-sion Adding WARCP to IVV enhanced the efficacy of IVV by boosting humoral and cellular immunity re-sponses with the advantages of high safety and dose-sparing.This study suggested the possibility of using WARCP as a novel immunologic adjuvant for influenza virus vaccine.

To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.

Background: New-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects. Objectives: In this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles. Methods: The mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination. Results: AEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4+ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months. Conclusions These findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.

Background: New-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects. Objectives: In this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles. Methods: The mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination. Results: AEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4+ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months. Conclusions These findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.