N-3 polyunsaturated fatty acids intake and risk of colorectal cancer: meta-analysis of prospective studies.

BACKGROUND: Growing body of laboratory evidence supports the beneficial effects of n-3 polyunsaturated fatty acids (PUFAs) on colorectal cancer (CRC) prevention. Epidemiologic studies investigating the relationship between n-3 PUFAs intake and risk of CRC, however, have been inconsistent. We aimed to clarify the relation by conducting a meta-analysis of prospective studies. METHODS: Eligible studies were identified by searching PubMed database and by carefully reviewing bibliographies of retrieved publications. Summary relative risks (RRs) with their 95 % confidence intervals (CIs) were computed with a random-effects model. Subgroup, meta-regression, and dose-response analyses were performed to explore potential sources of heterogeneity. RESULTS: A total of 14 prospective studies involving 8,775 cancer cases were included in the final analysis. Overall, total n-3 or marine PUFAs intake was not associated with risk of CRC (RR 0.99 and 1.00). However, there was a trend toward reduced risk of proximal colon cancer (total n-3 PUFAs: RR 0.83, 95 % CI 0.66-1.05; marine PUFAs: RR 0.81, 95 % CI 0.59-1.10) and a significant increased risk of distal colon cancer (total n-3 PUFAs: RR 1.26, 95 % CI 1.06-1.50; marine PUFAs: RR 1.38, 95 % CI 1.11-1.71). Furthermore, marine PUFAs intake accessed longer before diagnosis was associated 21 % reduced risk of CRC (RR 0.79, 95 % CI 0.63-1.00). CONCLUSION: Overall, this meta-analysis finds no relation between n-3 PUFAs intake and risk of CRC. The observed subsite heterogeneity within colon cancer and the possible effect modification by latency time merit further studies.

[Association of health literacy with health management among diabetics].

OBJECTIVE: To understand status of health literacy among diabetics and their health management behaviors, and analyze the relationship of health literacy and health management. METHODS: A two-staged cluster randomized sampling method was used to investigate 1 130 diabetics in Beijing, Ningbo and Xiamen from October to November in 2012. All participants should be diagnosed by primary hospital and above and have lived in the community over six months. Diabetic patients who indicated that they had severely impaired vision or cognitive disorder, or had severe physical deterioration, or did not live in the address provided were excluded. A total of 1 130 questionnaires were sent out and 1 083 eligible questionnaires were taken back, accounting for 96.87%. Multivariate logistic regression was adopted to analyze the association between health literacy and health management behaviors and blood glucose level. RESULTS: Among those participants, 47.7% (517) were men, 52.3% (566) were women, the age was (67.0 ± 9.5). According to diabetes health literacy scores, 73.7% (798/1 083) of them were classified as poor health literacy and 26.1% (283/1 083) as essential health literacy. Health literacy was associated with health management behaviors independently, demonstrating that the probability of utilizing health education, free physical examination, lifestyle guidance, monitoring blood glucose on their own, measuring blood glucose more than once a week and taking hypoglycemic agent regularly among diabetics with essential health literacy were 1.40 (95%CI:1.03-1.91), 1.65 (95%CI: 1.19-2.28), 2.70 (95%CI:1.98-3.69), 2.05 (95%CI:1.34-3.15), 2.56 (95%CI:1.85-3.56) , 1.48 (95%CI:1.07-2.06) times of those in diabetics with poor health literacy (P < 0.05). CONCLUSION: Health literacy may affect health management behaviors among diabetics. More activities targeted on diabetics with low health literacy were suggested to improve their' health literacy and their skills about diabetes mellitus management.

Comprehensive analysis of single-cell RNA and bulk RNA sequencing based on M2 tumor-associated macrophage and angiogenesis-related genes to assess prognosis and therapeutic response in lung adenocarcinoma.

M2 tumor-associated macrophage (M2 TAM), a crucial component of the tumor microenvironment, has a significant impact on tumor invasion and metastasis in the form of angiogenesis for lung adenocarcinoma (LUAD). In this study, both single-cell RNA and bulk RNA sequencing data were analyzed to identify 12 M2 TAM and angiogenesis-related genes (OLR1, CTSL, HLA-DPB1, NUPR1, ALOX5, DOCK4, CSF2RB, PTPN6, TNFSF12, HNRNPA2B1, NCL, and BIRC2). These genes were used to construct a prognostic signature, which was subsequently validated using an external cohort. Moreover, the immune profile analysis indicated that the low-risk group exhibited a distinct immune cell infiltration and relatively active status. Importantly, the prognostic signature was closely associated with PD-1, CTLA4, tumor mutation burden, and anti-cancer drug sensitivity. In summary, this study proposes a new prognostic signature for patients with LUAD based on M2 TAM and angiogenesis-related genes. The signature forecasts the prognosis of LUAD by an independent manner, reveals the potential molecular mechanisms involved in tumor immune-related functions, and offers appropriate clinical strategies for the treatment of patients with LUAD.

Novel insights into causal associations of body mass index or height with pneumothorax: a two-sample Mendelian randomization study.

Background: Observational studies have reported an association between body mass index (BMI) as well as height and the risk of pneumothorax. However, it has long been unclear whether BMI or height are causally associated with pneumothorax. Methods: Genetic summary data for BMI, height and pneumothorax were retrieved from multiple independent large genome-wide association studies (GWAS). A series of quality control steps were conducted to select instruments. Four independent two-sample Mendelian randomization (MR) analyzes were performed to adequately assess the causal relationship between BMI or height on pneumothorax, and the robustness of the results was assessed by a series of sensitivity analyzes. Results: Height increased the risk of pneumothorax with an OR of 1.5181 (95%CI 1.3092-1.7604; p = 3.28e-08); there was no evidence of a causal effect of BMI on the risk of pneumothorax with an OR of 0.8979 (95%CI 0.7417-1.0869; p = 0.269). Height increased the risk of spontaneous pneumothorax with an OR of 1.0010 (95%CI 1.0002-1.0018; p = 0.012); the results showed no significant causal relationship between BMI and spontaneous pneumothorax either with an OR of 0.9992 (95%CI 0.9983-1.0002; p = 0.112). Conclusion: Our results supported a genetic association between height and pneumothorax. We found that height increased the risk of pneumothorax. However, no evidence was found to suggest a causal relationship between BMI and pneumothorax risk. The relationship between BMI and pneumothorax requires further in-depth analysis.

[Pollution Characteristics and Risk Assessment of Heavy Metals in Surface Dusts and Surrounding Green Land Soils from Yellow River Custom Tourist Line in Lanzhou].

In order to investigate the contamination levels of dust and its surrounding green land soil heavy metal pollution and potential ecological and health risks in the scenic areas of urban waterfront parks, the gardens, squares, and theme parks of the Yellow River Custom Tourist Line in Lanzhou were selected as the research area, using 27 dust samples and 26 soil samples from its surrounding green lands. The contamination characteristics and potential ecological risks of eight heavy metals (Cr, Ni, Cu, Zn, As, Cd, Hg, and Pb) were evaluated using the geo-accumulation index (Igeo), single-factor pollution index (Pi), Nemerow integrated pollution index (PN), and improved potential ecological risk index (RI). The human health risk assessment was also evaluated using the exposure risk model. The results showed that the average concentrations of the other heavy metals in the surface dusts were higher than the background values of Gansu Province and Lanzhou City, except that the As mean concentrations in the surface dusts and the surrounding green land soils were slightly lower than the Gansu Province background values. For its surrounding green land soils, the mean concentrations of the other heavy metals (Cu, Zn, Cd, Hg, and Pb) exceeded the soil background values of Gansu Province and Lanzhou City, whereas the Cr and Ni mean concentrations were lower than their corresponding soil background values of Gansu Province and Lanzhou City. The geo-accumulation and single-factor pollution indices demonstrated that a slight to moderate pollution of Cr, Cu, Zn, Cd, Hg, and Pb occurred in surface dusts, and Cu, Zn, Cd, Hg, and Pb appeared in varying degrees of contamination levels in its surrounding green land soils. The Nemerow integrated pollution index analysis manifested that the overall contamination status of the study areas was between slightly and heavily polluted. The potential ecological risk index suggested that Cd and Hg were recognized as significant pollutant elements and that the RI of the other heavy metals were all below 40, presenting slight ecological risk. The health risk assessment indicated that ingestion was the dominant exposure pathway for heavy metals from the surface dusts and the surrounding green land soils, and no carcinogenic and noncarcinogenic risks posed threats to adults and children.

A predictive molecular signature consisting of lncRNAs associated with cellular senescence for the prognosis of lung adenocarcinoma.

The role of long noncoding RNAs (lncRNAs) has been verified by more and more researches in recent years. However, there are few reports on cellular senescence-associated lncRNAs in lung adenocarcinoma (LUAD). Therefore, to explore the prognostic effect of lncRNAs in LUAD, 279 cellular senescence-related genes, survival information and clinicopathologic parameters were derived from the CellAge database and The Cancer Genome Atlas (TCGA) database. Then, we constructed a novel cellular senescence-associated lncRNAs predictive signature (CS-ALPS) consisting of 6 lncRNAS (AC026355.1, AL365181.2, AF131215.5, C20orf197, GAS6-AS1, GSEC). According to the median of the risk score, 480 samples were divided into high-risk and low-risk groups. Furthermore, the clinicopathological and biological functions, immune characteristics and common drug sensitivity were analyzed between two risk groups. In conclusion, the CS-ALPS can independently forecast the prognosis of LUAD, which reveals the potential molecular mechanism of cellular senescence-associated lncRNAs, and provides appropriate strategies for the clinical treatment of patients with LUAD.

[Dynamic observation on MDA, iNOS and antisuperoxide anion in the liver of rats induced by acute nickel carbonyl poisoning].

OBJECTIVE: To investigate the level of malondiadehyde (MDA), antisuperoxide anion and inducible nitric oxide synthase (iNOS) in liver of rats poisoned by nickel carbonyl in order to discuss the mechanism of acute nickel carbonyl poisoning. METHODS: Healthy SD rats were intoxicated acutely by different concentrations of nickel carbonyl (20, 135 and 250mg/m3 for low, middle and high dose groups, respectively). SD rats inhaled by chlorine (250mg/m3 for chlorine group) were used as positive control group and other healthy SD rats as normal control group. Liver of animals was taken at different time points after exposure. The levels of MDA, iNOS and antisuperoxide anion were detected by biochemical assay. RESULTS: The contents of MDA and antisuperoxide anion in the liver of high dose group were significantly higher than that of other exposed groups and control group (P < 0.01). The contents of iNOS in middle and high dose group were higher than that in low dose group and control group (P < 0.05). CONCLUSION: The oxidative damage in the liver of SD rats could be induced by carbonyl nickel in air with increasing concentrations and in an obvious dose-response relationships.

[The sonodynamic effects of Chlorin e6 on the proliferation of human lung adenocarcinoma cell SPCA-1].

BACKGROUND AND OBJECTIVE: Sonodynamic therapy (SDT) is a promising new approach for cancer therapy. The purpose of this study is to detect the effects of SDT on the cell proliferation of human lung adenocarcinoma cell SPCA-1, using Chlorin e6 as a sonosensitizing agent activated by ultrasound. METHODS: SPCA-1 and normal peripheral mononuclear cell (PMNC) were treated with ultrasound or Chlorin e6 alone and combined. Cell proliferation was determined by MTT assay, and cell morphology was studied by inverted microscope after 6 h treated. RESULTS: 1.0 MHz ultrasound (1.0 W/ cm(2)-2.0 W/cm2 x 60 s) and Chlorin e6 (0.4 mg/mL-3.2 mg/mL) inhibited the cell proliferation of both SPCA-1 and PMNC cells in a intensity- and a dose-dependent manner respectively. Compared with the ultrasound (1.0 W/cm2 x 60 s) or Chlorin e6 (0.05 mg/mL-0.2 mg/mL) alone, the inhibitory effect on the cell proliferation was remarkably increased by the combination of ultrasound and chlorin e6 in SPCA-1 cells (P < 0.05), but no same effect was observed in PMNC cells (P > 0.05). Compared with the ultrasound (1.0 W/cm2 x 60 s) or chlorin e6 (0.2 mg/mL) alone, the combination treatment of ultrasound with Chlorin e6 induced more necrotic cells in SPCA-1 cells (P < 0.05). CONCLUSION: There was a significant selectively inhibitory effect of sonodynamic effect with Chlorin e6 on the SPCA-1 cell growth. Chlorin e6 may be a promising sonosensitizing agent for the treatment of non-small cell lung cancer.

[Multi-center phase II clinical trial of humanized anti-epidermal factor receptor monoclonal antibody h-R3 combined with radiotherapy for locoregionally advanced nasopharyngeal carcinoma].

OBJECTIVE: To evaluate the efficacy and safty of the humanized anti-epidermal factor receptor monoclonal antibody h-R3 in combination with radiotherapy for locoregionally advanced nasopharyngeal carcinoma. METHODS: Totally, 137 patients from 7 medical center around China were randomly divided into combined therapy group or control group. There was no difference in Karnofsky performance score between two groups. All patients in both groups received radical conventionally fractionated radiotherapy to the total dose of D(T) 70-76 Gy. For the combined therapy group, h-R3 was added at a dose of 100 mg i.v. weekly for 8 weeks started at the beginning of radiotherapy. RESULTS: Of the 137 eligilbe patients, 70 were in the combined therapy group treated by h-R3 plus radiotherapy and 67 in the control group by radiotherapy alone. The intent-to-treat (ITT) population consisted of 130 patients, while the per-protocol (PP) population was composed of 126 patients. The efficacy was assessed respectively at three point of time: the end of treatment, the 5th- and 17th-week after treatment. The complete response (CR) of the combined therapy group was significantly higher than that of the control group in both ITT and PP (ITT: 65.63%, 87.50%, 90.63% versus 27.27%, 42.42%, 51.52%; PP: 67.21%, 90.16%, 93.44% versus 27.69%, 43.08%, 52.31%; P < 0.05, respectively). The most common h-R3-related adverse reactions were fever (4.3%), hypotension (2.9%), nausea (1.4%), dizziness (2.9%) and rash (1.4%), which could be reversible if treated properly. Radiotherapy combined with 100 mg h-R3 i. v. weekly was tolerable and did not aggravate the side effects of radiation. The quality of life in the combined therapy group was comparable to that in the control group. CONCLUSION: This phase 1 multicenter clinical trial shows that h-R3 in combination with radiotherapy is effective and well-tolerated for the treatment of locoregionally advanced nasopharyngeal carcinoma.

[Clinical investigation of IRESSA in the treatment of patients with advanced refractory non-small cell lung cancer].

BACKGROUND: Chemotherapy is a main method for patients with advanced non-small cell lung cancer (NSCLC). NSCLC is usually a drug-resistant neoplasm. Innate or acquired drug-resis-tance contributes to the chief cause for bad effect in the treatment of patients with NSCLC. To search for a new anti-cancer drug becomes a goal of clinical oncologists. The aim of the present study is to evaluate the curative effect and side reactions of IRESSA in the treatment of patients with advanced refractory NSCLC. METHODS: The curative investigation was carried out after 100-day oral IRESSA by a dosage of 250mg/d in patients with advanced refractory NSCLC. The patients had ever experienced at least one regimen of chemotherapy. RESULTS: Totally 33 patients enrolled in this study and all were stage IV. There were 25 males and 8 females. All enrolled patients except one patient who died of severe adverse side reaction completed treatment by IRESSA. Thirty-two cases were evaluated. Complete response was obtained in 1 patient (3.1%). Partial response was seen in 11 patients (34.4%). The overall effective rate was 37.5% (12/32). The disease-control rate was 65.6% (21/32). Time to progression was 5.7 months. Overall survival time was 3.3 to 25.9 months (median survival time was 9.6 months). One-year survival rate was 28.1% (9/32). Two-year survival rate was 6.3% (2/32). The longest survivor lived for 25.9 months. The curative effect was correlated with the pathological type, in sequence of alveolar cell carcinoma, adenocarcinoma and squamous cell carcinoma. Almost all the adverse reactions were acceptable. The main adverse reactions included rash, itching of skin, arthralgia, diarrhea, anorexia, nausea, vomiting, dizziness, headache, chest distress and abdominal pain. No patients showed abnormal in liver or kidney function. No electrocardiogram abnormality was found. One patient who had chronic pulmonary fibrosis before died of respiratory failure due to severe interstitial pneumonia. CONCLUSIONS: IRESSA takes better effect on the advanced drug-resistant patients with NSCLC. So IRESSA may be accepted as third line in the treatment of advanced NSCLC and as first line in the treatment of patients with bad constitution who have no opportinities for operation, irradiation therapy or chemotherapy.

The detection of phytohemagglutinin-lymphokine activated killer cells' in vitro antitumor action with MTT colorimetry.

OBJECTIVE: To detect the in vitro antitumor action of phytohemagglutinin and lymphokine activated killer (PHA- LAK) cells. METHOD: MTT colorimetric assay was used to detect the in vitro cytotoxicity of PHA-LAK cells on K562 MGc80-3 143TK Hela and LoVo. RESULTS: The significant cytotoxicity of PHA-LAK cells on these five tumor cells from different organs could be found in vitro. The PHA-LAK cell activity on 143TK could reach 57.3% when the ratio of effective cell (EC) to target cell (TC) was 7.5:1. The antitumor effect did not increased or even decreased when the ratio of EC to TC was 15:1. CONCLUSIONS: (1)PHA-LAK cell has non-specific cytotoxicity against tumor cells and can overcome the problems of the quantity and activity of immunocyte of traditional adoptive cellular immunotherapy. (2)Under some conditions does MTT colorimetric assay be a susceptible, simple and convenient method for detecting the cytotoxicity of immunocytes.

[Dendritic cells transfected with carcinoembryonic antigen-vaccinia recombinant virus induces CEA-specific immunity mediated by cytotoxic T lymphocytes in vitro].

OBJECTIVE: To observe whether dendritic cells (DCs) transfected with carcinoembryonic antigen (CEA)-vaccinia recombinant virus (rV-CEA) induces cytotoxic T lymphocyte-mediated CEA-specific immunity in vitro. METHODS: DCs derived from peripheral blood monocytes were transfected with rV-CEA and then cocultured with autologous T cells. The proliferation of the induced T cells and their cytotoxic activity against CEA-secreting tumor cells were assessed and compared with those of T cells induced by DCs without rV-CEA transfection. RESULTS: T cells induced by DCs transfected with rV-CEA showed specific cytotoxicity against CEA-secreting tumor cells. CONCLUSION: DCs transfected with rV-CEA can elicit activation of CEA-specific T cells.

[Rapid serum-free culture of dendritic cells from human peripheral blood monocytes and their intracellular signal transduction].

OBJECTIVE: To explore the methods for rapid in vitro culture of the dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) under serum-free conditions and ascertain whether intracellular signal transduction pathway differs between calcium ionophore (CI) and tumor necrosis factor (TNF)- alpha during their induction of dendritic cell differentiation. METHODS: PBMCs isolated from healthy donors were plated in serum-free medium supplemented with 50 ng/ml rhGM-CSF. Cells cultured overnight were induced to differentiate with 100 ng/ml A23187 or 50 ng/ml TNF-alpha, given before or 30 min after pre-treatment with 0.5 mug/ml cyclosporine A (CsA). After culture for 40 h, the cell morphology was observed under phase-contrast microscope, and the surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was employed to assess the proliferation of the allogeneic T cells. RESULTS: PBMCs of healthy donors treated with 50 ng/ml rhGM-CSF in combination with 100 ng/ml CI or 50 ng/ml TNF-alpha for 40 h exhibited typical morphology of DCs with rapidly decreased CD14 expression and increased expressions of CD83 and co-stimulatory molecules (CD80 and CD86), showing also enhanced ability of stimulating allogeneic T cell proliferation. Calcineurin antagonist CsA inhibited the differentiation induced by CI, but not that induced by TNF-alpha. CONCLUSIONS: Under serum-free conditions, both CI and TNF-alpha are capable of inducing rapid DC differentiation from human PBMCs, but the intracellular signal transduction of CI-induced differentiation is different from that induced by TNF-alpha.

[Effect of immunocyte therapy on benzene-induced bone marrow haemopoietic dysfunction].

OBJECTIVE: To explore the effect of treatment with immunocyte therapy on benzene-induced haemopoietic dysfunction. METHODS: Mono-nuclear cells (MNC) were separated from 40 - 50 ml peripheral blood in patients and mixed with interleukin-2 and granulocyte macrophage colony stimulating factor (GM-CSF) for six day cultivation. The new formed immunocytes were collected and transfused into the patients. Bone marrow aspiration and biopsy were taken before and after therapy for all patients with severe benzene poisoning. Blood samples were stained by flow cytometry for detecting CD(4) and CD(8) positive cells. RESULTS: Of 20 patients with chronic benzene poisoning, 9 were severe benzene poisoning. All examination including blood count, bone marrow biopsy and T cell subpopulation restored to normal after immunocyte therapy. Laboratory tests (liver and kidney function, and myocardial enzymes) were observed periodically and showed normal during therapy. Follow-up study (the longest time was more than 15 months) showed that bone marrow haemopietic function of all treated patients were in normal range. CONCLUSION: Bone marrow haemopoietic dysfunction caused by benzene poisoning may be closely related to disorder of immune function. Immunocyte therapy may significantly improve bone marrow haemopoietic dysfunction induced by benzene poisoning.

[Adriamycin enhances the sonodynamic effect of chlorin e6 against the proliferation of human breast cancer MDA-MB-231 cells in vitro].

OBJECTIVE: To evaluate the effect of adriamycin (ADM) in enhancing the sonodynamic effect of chlorin e6 against the proliferation of human breast cancer MDA-MB-231 cells in vitro. METHODS: MDA-MB-231 cells were treated with ultrasound/Chlorin e6 alone or in combination with ADM, and the changes in the cell proliferation was determined by MTT assay. RESULTS: Ultrasound (1.0 MHz) at the power intensity of 0.5-2.0 W/cm2 inhibited the proliferation of MDA-MB-231 cells in an intensity-dependent manner, and chlorin-e6 (0.05-1.6 mg/ml) and ADM (0.1-0.4 g/ml) alone both inhibited the proliferation of MDA-MB-231 cells dose-dependently. Compared with ultrasound (0.5 W/cm2, 1.0 MHz, 60 s) or chlorin-e6 (0.05-0.2 mg/ml) alone, a combined treatment with ultrasound and chlorin e6 significantly enhanced the inhibitory effect on the proliferation of MDA-MB-231 cells (P<0.05). ADM significantly enhanced the sonodynamic effect of chlorin e6 (0.1 mg/ml) against the cell proliferation of MDA-MB-231 cells (P<0.05), and the effect was schedule-dependent, which was greater when ADM was added after the sonodynamic treatment (P<0.05). CONCLUSION: ADM can enhance the sonodynamic effect of chlorin e6 against the proliferation of MDA-MB-231 cells in vitro.

Sonodynamic and photodynamic therapy in advanced breast carcinoma: a report of 3 cases.

Photodynamic therapy (PDT) is an established therapeutic method, first approved by the FDA for certain kinds of cancer in 1998. There are also increasing data to show that a related procedure, sonodynamic therapy (SDT), is a promising new modality for cancer treatment. Here, the authors report clinical results in 3 advanced refractory breast cancer patients who were treated using a combination of sonodynamic and photodynamic therapy (SPDT), along with conventional therapies. All 3 patients had pathologically proven metastatic breast carcinoma. These widely disseminated carcinomas had ultimately failed to respond to conventional therapy. A new sensitizing agent, Sonoflora 1 (SF1) was administered sublingually; then, after a 24-hour delay, patients were treated with a combination of light and ultrasound. All patients had significant partial or complete responses. SPDT is a promising new therapeutic combination for the treatment of breast cancer.

Ex vivo activated immune cells promote survival and stimulate multilineage hematopoietic recovery in myelosuppressed mice.

Recovery of multilineage hematopoiesis from severe myelosuppression due to chemotherapy and radiotherapy remains a clinical problem. The authors have developed a simple immunotherapy to treat this disease in a mouse model. Syngeneic spleen cells or xenogeneic human peripheral mononuclear cells were cultured ex vivo with a combination of IL-2 at 500 IU/mL, GM-CSF at 200 U/mL, and calcium ionophore A23187 at 100 ng/mL for 2 days and injected intravenously into mice that had previously received a lethal dose of carboplatin and radiation. The therapy was highly effective: a single injection of activated cells enhanced survival and simulated multilineage recovery of hematopoiesis. Ex vivo activated immune cells produced multiple cytokines, including several hematopoietic growth factors. Adherent cells were found to be more potent than nonadherent cells in promoting survival, and the therapy alone was capable of mobilizing peripheral blood stem cells in normal mice.

[The effect of calcium ionophore on dendritic cells derived from peripheral blood mononuclear cells].

AIM: To explore the effect of calcium ionophore (CI) on dendritic cells (DC) derived from peripheral blood monocytes. METHODS: Peripheral blood monocytes from healthy donors were treated with 100 microg/L rhGM-CSF, 100 microg/L rhGM-CSF plus (10 microg/L) CI, and 100 microg/L rhGM-CSF plus (100 microg/L) respectively. After cultivation of 40 hours, cellular morphology was observed under light microscope and electron microscope. Surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was used to detect proliferation of autologous T cells. RESULTS: Peripheral blood monocytes treated with 100 microg/L rhGM-CSF plus 100 microg/L CI for 40 hours showed typical morphology of DCs. Simultaneously there was a decrease in CD14 expression and increase in HLA-DR, CD40, CD83 and CD86 expressions on these cells. In addition, PBMCs treated with 100 microg/L rhGM-CSF pass 100 microg/L CI for 40 hrs. Could evidently stimulate proliferation of autologous T cells. CONCLUSION: CI can markedly enhance transformation of peripheral blood monocytes induced by GM-CSF into DCs.

Ex vivo immunotherapy for patients with benzene-induced aplastic anemia.

Aplastic anemia is a bone marrow failure disorder characterized by pancytopenia and a hypocellular marrow. Benzene is one of the etiologic agents capable of inducing the disease. With modest to severe aplastic anemia, one previously untreated patient and 13 patients who had failed immunosuppressive therapy were studied. Peripheral blood mononuclear cells from patients were expanded in vitro with a combination of cytokines and a calcium-mobilizing agents for 2 days, and the activated cells were infused intravenously once a week. In some cases, we used allogenic leukocytes instead of autologous cultured lymphocytes. After 6-35 weeks of the treatment, all patients had multilineage responses to this therapy and achieved complete disease remission, defined as normal blood count, independence from transfusion, and normal bone marrow histology. The therapy was safe and well tolerated with minimal side effects. The cultured cells produced interleukin-1 and induced immune responses in vivo. Serum interleukin-2 and interferon- gamma were detected following cell infusion. Finally, patients had sustained responses to the therapy and no relapse was found up to 18 months after cellular therapy.

[Signal transduction in the differentiation of human peripheral blood mononucleocytes towards dendritic cells induced by calcium ionophore].

AIM: To explore the intracellular signal transduction pathway in the differentiation of human peripheral blood mononucleocytes (PBMCs) towards dendritic cells (DCs) induced by calcium ionophore (CI). METHODS: PBMCs isolated from a healthy donor were cultured with A23187 plus rhGM-CSF, 100 microg/L each. In some experiments, PBMCs were cultured for 30 minutes with W-7 (10 micromol/L), CsA(0.5 mg/L) or KT5926(1 micromol/L) before addition of rhGM-CSF and A23187. After culture for 40 hours, morphological change of the cells were observed under phase contrast microscope; surface markers on treated PBMCs were analyzed by flow cytometry; the proliferation of allogeneic human T cells stimulated by the treated PBMCs was detected by MTT colorimetry. RESULTS: PBMCs of the healthy donor cocultured with rhGM-CSF plus CI for 40 hours had the typical morphology of DCs, with decreased CD14 expression, and increased CD83, CD80 and CD86 expressions. The proliferation of allogeneic T cells stimulated by PBMCs treated with A23187 plus rhGM-CSF was strengthened. But the morphological changes, surface marker expressions and the ability to enhancing proliferation of allogeneic T cells were inhibited to different degrees by W-7, CsA or KT5926. CONCLUSION: The differentiation of PBMCs towards DCs by CI may be modulated by Ca (2+)/calmodulin and multiple signal transduction pathways downstream.