[The current situation of occupational burnout and its influencing factors among orphan child care workers in Nanjing].

Objective: To explore the status of job burnout of nurses for orphans and disabled children in Nanjing and its influencing factors, so as to provide reference for promoting the mental health of nurses for orphans and disabled children. Methods: From February to May, 2017, a cross-sectional survey was conducted among 236 nurses working in social welfare institutions for orphans and disabled children in Nanjing, using Occupational Stress Indicator (OSI) and Maslach Burnout Inventory (MBI) . It investigated the status of job burnout among nurses of orphans and disabled children, logistic regression analysis was used to explore the effect of occupational stress on the incidence of job burnout of nurses for orphans and disabled children. Results: The results showed that the incidence of job burnout was 67.37% (159/236) , the incidence of job burnout was negatively correlated with behavioral characteristics and family support, the incidence of emotional exhaustion was positively correlated with task control and job monotony (P<0.05) , and negatively correlated with role ambiguity and job satisfaction (P<0.05) . The incidence of depersonalization was negatively correlated with job monotony and peer support (P<0.05) . Conclusion: The nurses of orphans and disabled children have serious job burnout. Occupational stress factors, personality characteristics and relieving factors have influence on the incidence of job burnout.

[Study on occupational stress response of orphans and disabled children care workers in Nanjing].

Objective: To explore the status quo of occupational stress and its influencing factors of nursing staff for orphaned and disabled children in Nanjing, and to put forward measures for adverse reactions to occupational stress, so as to ensure the psychological health of this occupational population. Methods: From February to May 2017, 236 nursing assistants for orphans and disabled children in Nanjing social welfare home were selected by cluster sampling method. The occupational stress index(occupational stress indictor, OSI) was used to investigate occupational stress response and occupational stress factors. Partial correlation and multiple linear regression were used for statistical analysis. Results: The scores of job satisfaction, mental health and depressive symptoms in occupational stress reaction were (43. 99±6. 83) , (36. 09±4. 59) and (17. 31±2. 44) re- spectively. In terms of job satisfaction, work monotony, logic and compound change were the contributing fac- tors (P<0. 05) , task strategy and task control were negative factors (P<0. 05) ; on mental health, opportunities were raised and participation in decision-making Self-esteem, technology utilization, environmental control, time management, task strategy and support of colleagues as contributing factors (P<0. 05) , ambition and role conflicts as negative factors (P<0. 05) ; on depression, work input, participation Decision-making, promotion opportunities and behavioral characteristics were protective factors (P<0. 05 ) . Conclusion: Occupational stress among caregivers of orphans and disabled children cannot be ignored. Occupational stress reaction is serious. Occupational stress factors should be reduced and individual stress coping ability should be enhanced.

[Analysis of occupational stress and occupational exhaustion of primary caregivers].

Objective: To investigate the occupation stress and job burnout of orphan child care workers in Nanjing and nursing staff in Yangzhong primary hospital, to compare the differences of occupational stress and job burnout between the two social service workers. Methods: From February to May 2017, a cross- sectional survey was conducted. Cluster sampling method was used to investigate the occupation stress and job burnout of orphan child care workers in Nanjing and nursing staff in Yangzhong primary hospital, totally 403 people, by a combination of the Internet of things assessment system of occupational stress and traditional questionnaire. Results: There were statistically significant differences of occupational stress and job burnout scores of both social service workers (P<0.05). Orphan child care workers in job control, technology utilization, job routinization, work risk, role ambiguity and participation decision-making, anxiety state, body complain, life attitude, behavior characteristics, ambition, anxiety trait, social support, logic, family-work relationship and depersonalization were higher than those of Yangzhong primary nurses (P<0.05); Primary nurses in Yangzhong in quantitative load and change, workload, role conflict, job prospects, promotion, depressive symptom, daily stress, self-esteem, task strategy, time management, work input, and personal accomplishment reduction scored higher than those of Orphan child care workers (P<0.05). The logistic regression analysis of the occupational depletion positive rate of the two social service personnel showed that the protective factors of the nursing staff of the orphans and disabled children had behavioral characteristics and family support(OR=0.877, P=0.044; OR=0.691, P=0.001) . The contributing factors of the primary nursing staff in Yangzhong were task control, workload, work psychological control source (OR=1.110, P=0.019; OR=1.128, P<0.001;OR=1.066, P=0.032) . The protection factors were technical utilization, opportunities for improvement, and mental health (OR=0.775, P=0.005; OR=0.765, P=0.006; OR=0.914, P=0.002) . Conclusion: The work content, the service object and the nature of the unit of primary caregivers influence the state of occupational stress and job burnout. The influencing factors of occupational stress and job burnout are not identical, Different social service personnel show occupational stress and occupational exhaustion in different aspects, and it is necessary to propose mitigation measures for two kinds of social service personnel.

CdS nanoparticles of different lengths induce differential responses in some of the liver functions of mice.

Cadmium sulfide nanoparticles (CdS NPs) are one of important nanoparticle materials which are widely used in photoelectric production, but their potential health hazard to the liver is not clear. This study is aimed at exploring the possible mechanisms of liver injury induced by CdS NPs. Male mice were treated with nanoparticles of 110-130 nm and 80-100 nm cadmium sulfide. The main methods were based on detecting the vigor of superoxide dismutase (SOD) and glutathione (GSH), and content of malondialdehyde (MDA) in both blood and liver tissues as well as on observing the pathological changes in liver tissue. CdS NPs suppressed the activity of SOD and GSH, and increased the serum MDA content (p < 0.05); both effects were observed together in liver tissues of 80-100 nm group (p < 0.05) and were accompanied by an obviously inflammatory response. CdS NPs induced oxidative damage and inflammatory response in liver tissue, which may be an underlying mechanism for its pulmonary toxicity. Additionally, the toxicity of CdS NPs was closely related to the size of nanoparticles. Pathological results showed that the hepatotoxicity of shorter CdS NPs is greater than that of longer CdS NPs (Tab. 6, Fig. 3, Ref. 20).

[The influence of demographic characteristics on occupational stress of orphan child care workers].

Objective: To investigate the occupation stress of orphan child care workers in Nanjing, to explore the influence of different demographic characteristics on occupational stress of orphan child care workers and propose measures. Methods: Cluster sampling method was used to investigate the occupation stress of 236 people of orphan child care workers in Nanjing by a combination of the Internet of things assessment system of occupational stress and traditional questionnaire. Results: The scores of workload, job prospects, participation decision-making, anxiety and logical in different age groups among orphan child care workers were statistically different (P<0.05) ; The scores of organizational loyalty in different education groups among orphan child care workers were statistically different (P<0.05) ; The scores of task control, resource control, workload, job prospects, job satisfaction, mental health, anxiety and ambition were statistically different among orphan child care workers with different job titles (P<0.05) ; The scores of role conflict, role ambiguity, job satisfaction, mental health, anxiety, self-esteem, colleague support, family support and task strategy were statistically different orphan child care workers in different classes (P<0.05) . Conclusion: Different demographic characteristics will have certain influence on occupational stress of orphan child care workers.

[Impacts of EGFR 19 exon mutations on brain metastases in treatment-naïve patients with lung adenocarcinoma].

Objective: To investigate the relationship between the status of epidermal growth factor receptor (EGFR) mutations and brain metastases in patients with lung adenocarcinoma. Methods: From August 2010 to May 2015, a total of 1 063 lung adenocarcinoma patients with identified status of EGFR mutations in Shanxi Cancer Hospital were enrolled, of which 456 patients had EGFR mutations. Multivariate Logistic regression model was used to analyze the correlation between EGFR mutation status and brain metastases in patients with lung adenocarcinoma. Results: In 125 patients with brain metastases before initial treatment, 65 patients had EGFR mutations, including 36 patients with deletion mutations in exon 19. The frequency of EGFR 19 exon mutation was 28.8% (36/125). Among 456 patients with EGFR mutations, 65(14.3%) patients were with brain metastases, in which 36(55.0%) had deletion mutations in exon 19. The multivariate analysis showed that age, Eastern Cooperative Oncology Group (ECOG) score, EGFR mutations and N staging were associated with brain metastases(P<0.05). Further subgroup multivariate analyses showed that age, ECOG score, mutation status in exon 19 and N staging were associated with brain metastases (P<0.05). Conclusions: EGFR mutation status is related to brain metastases. Mutations in EGFR exon 19 is an independent risk factor for brain metastases.

Effects of ovarian ablation or suppression in premenopausal breast cancer: A meta-analysis of randomized controlled trials.

BACKGROUND: The effect of ovarian ablation or suppression (OAS) in premenopausal women with breast cancer is controversial. The overall survival (OS), disease-free survival (DFS) and adverse event of OAS versus no OAS were compared. METHODS: A literature review of EMBASE, Web of Science, PUBMED, and Cochrane Library was conducted. The hazard ratio (HR) and 95% confidence interval (CI) for OS and DFS, as well as risk ratio (RR) and 95% CI for adverse events were evaluated. I-squared statistic (I2) represents heterogeneity. RESULTS: Twenty-nine studies with a total of 21,249 women were included. In premenopausal women aged 40 years or younger, there were significant differences in OS (HR 0.78, 95% CI: 0.66-0.94, P=0.008, I2 = 0%) and DFS (HR 0.84, 95% CI: 0.73-0.97, P=0.02, I2 = 0%) between OAS and no OAS. In advanced stage breast cancer, a significant difference was found in OS (HR 0.76, 95% CI: 0.60-0.96, P=0.02, I2 = 0%). Patients treated with OAS had more chances to have hot flushes (RR 1.91, 95% CI: 1.62-2.26, P < 0.01, I2 = 0%) and vaginal dryness (RR 1.19, 95% CI: 1.08-1.31, P=0.0003, I2 = 0%). No significant difference in depression (RR 1.28, 95% CI: 0.94-1.74, P=0.12, I2 = 0%). CONCLUSIONS: The study shows that OAS plays a beneficial role in premenopausal women aged 40 years or younger and advanced stage breast cancer. However, OAS is associated with increase in hot flushes and vaginal dryness.

Intratumor murine interleukin-12 gene therapy suppressed the growth of local and distant Ewing's sarcoma.

We evaluated the effect of interleukin-12 (IL-12) gene therapy using an Ewing's sarcoma animal model in T-cell-deficient nude mice. Subcutaneous injection of TC71 cells resulted in tumor development by day 5. Mice were treated with a single intratumor injection of adenovirus beta-galactosidase (Ad.beta-gal) or adenovirus murine IL-12 (Ad.mIL-12) (2 x 10(9) PFU) and killed 1-7 days later. Reverse transcriptase-polymerase chain reaction analysis of tumor tissue demonstrated peak expression of IL-12 p35 and p40 at 48 h, which persisted up to 7 days. For in vivo therapy, mice received intratumor Ad.beta-gal or Ad.mIL-12 twice weekly for 2.5 weeks starting on day 6. Ad.mIL-12-treated tumors were significantly smaller (median volume, 19.7 mm3; range, 3.41-159.5 mm3) than Ad.beta-gal-treated tumors (median volume, 3214.9 mm3; range 1679.9-5909.8 mm3, P<0.003) on day 31. The weight of Ad.mIL-12-treated tumors was also lighter than the Ad.beta-gal-treated tumors (median, 2 mg; range, 1-5 mg versus median, 1960 mg; range 1640-5230 mg, P<0.01). Ad.mIL-12 therapy significantly prolonged the survival time and also inhibited the growth of an untreated tumor on the contralateral side. Immunohistochemistry analysis of the IL-12-treated tumors demonstrated IL-12 expression with increased Fas, Fas ligand and tumor cell apoptosis. CD31 and vascular endothelial growth factor expression were decreased. These data suggest that IL-12 gene therapy may be useful in the treatment of Ewing's sarcoma.

Growth suppression of established human osteosarcoma lung metastases in mice by aerosol gene therapy with PEI-p53 complexes.

Lung metastases are a frequent complication of osteosarcoma and a treatment that would reduce the severity of this complication would be of great benefit to patients. We have used a formulation consisting of polyethyleneimine (PEI) and a p53 gene administered in aerosol to treat established lung micrometastases as a model of human osteosarcoma in nude mice. The SAOS-LM6 cell line, a metastatic derivative of the p53 null SAOS-2 line, expresses high levels of p53 protein after in vitro transfection with PEI-p53 complexes as determined by ELISA, and transfection with both p53wt and the p53 variant, p53-CD(1-366) in vitro, results in a marked inhibition of SAOS-LM6 cell proliferation. Aerosol delivery of plasmid DNA containing either the p53 gene or a p53-CD(1-366) variant gene formulated with PEI to mice resulted in highly significant reductions in the numbers and size of tumors (P<.001), the total number of tumor foci in the lungs (P<.001) and the size of individual tumor nodules in treated animals compared to untreated, PEI only-treated and PEI-CAT-treated control animals. The different tissues examined did not reveal any signs of toxicity or inflammation after repeated exposure to PEI-DNA. The aerosol delivery of PEI-based formulations of p53 or synthetic p53 variant genes represents a promising new strategy for the treatment of established human osteosarcoma lung metastases. The noninvasive nature of aerosol delivery coupled with low toxicity also make this therapeutic approach potentially appropriate for combination therapy with either radio- or chemotherapy.

Interleukin (IL)-12 and IL-12 gene transfer up-regulate Fas expression in human osteosarcoma and breast cancer cells.

Expression of Fas (CD95, APO-1), a cell surface receptor capable of inducing ligand-mediated apoptosis, is involved in tissue homeostasis and elimination of targeted cells by natural killer and T cells. Corruption of this pathway, such as reduced Fas expression, can allow tumor cells to escape elimination and promote metastatic potential. In this study, the status of Fas expression has been examined in the parental SAOS human osteosarcoma cells that do not metastasize and in selected variants that cause lung metastases in 16 weeks (LM2) or 8 weeks (LM6) after i.v. injection into nude mice. Fas expression correlated with the metastatic potentials of the three cell lines. Northern and fluorescence-activated cell-sorting analyses indicated that LM6 cells expressed Fas at a lower level than seen in the parental cells. Infection of the LM6 cells with an adenoviral vector containing the murine interleukin (IL)-12 gene (AD:mIL-12) or treatment with recombinant murine IL-12 resulted in a dose-dependent up-regulation of FAS: The up-regulation of Fas by IL-12 was also demonstrated in human etoposide-resistant MDA-MB-231 breast cancer cells. [(3)H]Thymidine growth inhibition studies indicated that the cell surface Fas induced after IL-12 exposure was functional and able to mediate cell death on cross-linking with anti-FAS: We also demonstrate that this effect is independent of IFN-gamma. Whereas these cell lines are sensitive to IFN-gamma, incubation with IFN-gamma does not increase susceptibility to Fas-mediated cell death, nor do these cells produce IFN-gamma with or without IL-12 treatment. We hypothesize that expression of Fas may play a role in the elimination of metastatic tumor cells in the lung, an organ in which Fas ligand is expressed. The antitumor activity of IL-12 may be secondary in part to its ability to up-regulate Fas expression on tumor cells, which subsequently increases immune-mediated destruction of osteosarcoma cells.

E1A sensitizes HER2/neu-overexpressing Ewing's sarcoma cells to topoisomerase II-targeting anticancer drugs.

Overexpression of the HER2/neu oncogene is associated with tumorigenicity and drug resistance in many types of cancer. Three different human Ewing's sarcoma cell lines (TC71, RD, and A4573) were found to express high levels of the HER2/neu protein. Transduction of TC71 cells with the E1A gene using an adenoviral vector (Ad-E1A) down-regulated HER2/neu overexpression in those cells and increased cytostasis. E1A-induced apoptosis was demonstrated by both flow cytometric analysis and Western blot analysis using a poly(ADP-ribose) polymerase antibody. After transduction of the E1A gene into these cells, the sensitivity of these cells to VP-16 (etoposide) was enhanced 18-fold and to Adriamycin 5-fold. However, no change was seen in cisplatin sensitivity. E1A also significantly increased topoisomerase IIalpha protein expression, indicating that the up-regulation of topoisomerase IIalpha may be one of the mechanisms by which E1A enhanced the sensitivity to topoisomerase II-targeting anticancer drugs, such as VP-16 and Adriamycin, but not cisplatin. In summary, these studies demonstrated that Ad-E1A can down-regulate HER2/neu overexpression and up-regulate topoisomerase IIalpha expression in human Ewing's sarcoma cells, increasing their apoptosis rate and enhancing their sensitivity to VP-16 and ADRIAMYCIN:

Intranasal therapy with an adenoviral vector containing the murine interleukin-12 gene eradicates osteosarcoma lung metastases.

The purpose of these studies was to determine the effect of adenovirus-mediated interleukin-12 (IL-12) gene transfer on the growth and development of osteosarcoma (OS) lung metastases in nude mice. A nude mouse model was produced by repetitive cycling of human SAOS OS cells through the lung. The resultant SAOS-LM6 cell line produced microscopic lung metastases by 5-6 weeks after i.v. injection of the tumor cells, with visible lung metastases present 8 weeks after injection. Transfection of SAOS-LM6 cells with a plasmid containing the murine IL-12 gene resulted in a decrease in metastatic potential. Animals injected with IL-12-transfected clones had fewer metastases compared with mice injected with SAOS-LM6 cells transfected with a control plasmid. Furthermore, nasal delivery of an adenoviral vector containing the murine IL-12 gene resulted in the inhibition of pulmonary metastases. Together, these data indicate that IL-12 may be an effective agent against OS and that nasal delivery may offer a unique way to deliver the gene to the local tumor environment, potentially decreasing systemic toxic effects.

9-Nitrocamptothecin liposome aerosol treatment of melanoma and osteosarcoma lung metastases in mice.

The response rates of relapsed osteosarcoma and melanoma pulmonary metastases to traditional i.v. chemotherapeutic regimens have been disappointing. Direct drug delivery of chemotherapy to the lungs could increase the drug concentration in the tumor area and may offer a new therapeutic approach for these patients. Previous studies demonstrated that drugs delivered to the respiratory tract in liposomal formulation resulted in high pulmonary drug concentration, reduced systemic toxicity, and reduced dosage requirements compared with parenteral and oral administration. To determine whether this approach has utility against pulmonary metastases, the efficacy of aerosol therapy with liposome-encapsulated 9-nitrocamptothecin (L-9NC) was determined using two different experimental lung metastasis models. C57BL/6 mice were treated the day after the i.v. injection of B16 melanoma cells with aerosol L-9NC for 1 h (153 microg 9-nitrocamptothecin/kg) for 5 days per week for up to 3 weeks. Aerosol L-9NC treatment resulted in a reduction in lung weights (P = 0.005) and number of tumor foci (P < 0.001). Visible tumor nodules were fewer and smaller in the 9-nitrocamptothecin-treated group than in untreated control mice (P < 0.001). Using a newly developed human osteosarcoma experimental metastasis model in nude mice, we demonstrated that aerosol L-9NC was also effective against established lung metastases. Aerosol therapy initiated on the ninth week after i.v. tumor injection and continued for 8 or 10 weeks produced highly significant reductions in the number of animals with both visible and microscopic disease (P < 0.02), the total number of tumor foci in the lungs (P < 0.005), and the size of the individual tumor nodules (P < 0.02). These data suggest that L-9NC aerosol therapy may offer significant advantage over existing methods in the treatment of melanoma and osteosarcoma pulmonary metastases.

ImmTher, a lipophilic disaccharide derivative of muramyl dipeptide, up-regulates specific monocyte cytokine genes and activates monocyte-mediated tumoricidal activity.

ImmTher, a liposome-encapsulated lipophilic disaccharide tripeptide derivative of muramyl dipeptide, previously showed activity against liver and lung colorectal metastases in a phase I trial. The purpose of the current studies was to investigate whether ImmTher could up-regulate specific cytokine gene expression and protein production, as well as activate the tumoricidal or cytostatic activity of human monocytes. ImmTher induced the expression and production of interleukin(IL)-1alpha IL-1beta, IL-6, IL-8, IL-12, macrophage chemotactic and activating factor, and tumor necrosis factor alpha but not IL-2 or IL-10. Cytostatic or cytotoxic monocyte activity was stimulated against human Ewing's sarcoma, osteosarcoma, and melanoma cells but not breast cancer cells. Production and secretion of these cytokine proteins may play a role in the antitumor activity of ImmTher.

Interferon-alpha enhances the sensitivity of human osteosarcoma cells to etoposide.

The purpose of these studies was to determine whether interferon-alpha (IFN-alpha) could enhance the sensitivity of human osteosarcoma cells to the cytotoxic actions of etoposide (VP-16). Cytostasis was determined using a [3H]thymidine incorporation assay, whereas cytotoxicity was quantified by a colony-formation assay. Low concentrations (0.1-5 U/ml) of IFN-alpha enhanced the cytostatic activity of VP-16 against MG-63, SAOS-2, and TE-85 osteosarcoma cells. The cytostatic activity of 1 microM VP-16 rose from 11% to 64%, 9% to 31%, and 10% to 71%, respectively, in the three cell lines when IFN-alpha was present. Survival fraction was also decreased when the osteosarcoma cells were treated with VP-16 + IFN-alpha as compared to either agent alone. The interaction between these two agents was determined to be synergistic rather than additive by interaction index analysis. Similar effects on cytostasis and cytotoxicity were observed when IFN-alpha was combined with Adriamycin but not cisplatin, actinomycin D, vinblastine, or amsacrine. VP-16 uptake was enhanced 12-fold in the presence of IFN-alpha, but this did not appear to translate into an increase in topoisomerase-II (topo-II)-DNA complex formation as quantified by the sodium dodecyl sulfate-KCl precipitation assay. We also could not detect alterations in topo-II expression, topo-II protein production, or cell cycle kinetics that have been shown to correlate with increased VP-16 cell sensitivity. Therefore, at this time the mechanism of enhanced cell sensitivity to the combination treatment remains unclear.

A nude mouse model of human osteosarcoma lung metastases for evaluating new therapeutic strategies.

The purpose of these studies was to develop a metastatic osteosarcoma nude mouse model to evaluate the in vivo efficacy of new therapeutic compounds. Human SAOS-2 osteosarcoma cells (10(6) cells) were injected i.v. into nude mice. Cells isolated from a rare pulmonary metastases 6 months later were established (SAOS-LM1) in culture and re-injected. This procedure was repeated 5 additional times to produce the SAOS-LM6 cell line. Visible pulmonary nodules were present 8 weeks following i.v. injection of 10(6) SAOS-LM6 cells as compared to 17 weeks using SAOS-LM2 cells. Microscopic SAOS-LM6 pulmonary metastases were demonstrated at 6 weeks. Administration of adriamycin on week 9 resulted in regression of macroscopic SAOS-LM6 lung tumors. The ability of the model to be used to evaluate the effectiveness of a biologic agent against microscopic disease was also verified. It was concluded that this model can assess therapeutic efficacy and therefore, may have a role in investigating the potential of novel approaches aimed at eliminating pulmonary metastatic osteosarcoma.

Phase II study of recombinant interleukin 1alpha and etoposide in patients with relapsed osteosarcoma.

A Phase II trial using interleukin 1alpha (IL-1alpha) and etoposide for patients with relapsed osteosarcoma (OS) was undertaken to assess the feasibility and tolerability of combination therapy with biotherapy and chemotherapy. Nine patients with histologically proven relapsed OS were treated with IL-1alpha immediately followed by etoposide daily for 5 days every 3 weeks. Surgical resection of lung metastasis or peripheral tumor was performed after two or three cycles. We observed three partial responses; disease was stable in another case. One case could not be evaluated. The side effects associated with combination therapy were as predicted from known side effects of the individual agents; however, more profound neutropenia was observed. Four patients exhibited clinical signs of capillary leak syndrome, i.e., hypotension, edema, and weight gain. The etiology of the capillary leak was unclear, because serum IL-1alpha, IL-2, tumor necrosis factor, and nitric oxide levels could not be used to predict which patients would develop capillary leak. Histological analysis of tumor specimens obtained after two or more courses of therapy showed changes consistent with a response to a biological response modifier: peripheral fibrosis surrounded the metastasis with infiltration of chronic and acute inflammatory cells. Because the response of relapsed OS to any type of salvage regimen has been poor, we interpret the clinical response of this therapy as good. However, the significant side effects associated with this therapy must also be taken into consideration before deciding to use this combination therapy. It is unfortunate that the study was stopped early due to halted production of IL-1alpha. If this agent is again manufactured for clinical use, we conclude that additional evaluation in patients with relapsed OS is warranted.

Altered monocyte chemotactic and activating factor gene expression in human glioblastoma cell lines increased their susceptibility to cytotoxicity.

A cDNA encoding for human monocyte chemotactic and activating factor (MCAF) was ligated into the retroviral vector pLXSN. These pMCAF-LXSN and antisense p-antiMCAF-LXSN vectors were transfected into HBT20 and HBT28 human brain tumor cells. HBT28 cells constitutively express high amounts of MCAF, whereas HBT20 cells express much less MCAF. HBT20 cells transfected with pMCAF-LXSN (HBT20-MCAF) showed significantly higher MCAF mRNA expression and MCAF protein production than the HBT20-parent or HBT20 cells transfected with control vector (HBT20-LXSN). In contrast, supernatant from HBT28 cells transfected with p-antiMCAF-LXSN (HBT28-antiMCAF) contained less MCAF than HBT28-parent, HBT28-LXSN, and HBT28-MCAF cells. Activated human monocytes killed HBT20-MCAF cells more efficiently compared with HBT20-parent, HBT20-LXSN, and HBT20-antiMCAF cells (P< 0.02), whereas HBT28-antiMCAF cells were killed more efficiently by activated monocytes compared with HBT28-parent, HBT28-LXSN, and HBT28-MCAF cells (P< 0.05). Cultured supernatants from activated monocytes plus HBT20-MCAF cells or from activated monocytes plus HBT28-antiMCAF cells inhibited the growth of HBT20 and HBT28 cells, respectively. Altered MCAF expression can therefore enhance the ability of activated monocytes to kill brain tumor cells. This increased cytotoxicity is partially dependent upon the basal state of MCAF in the individual tumor cells.

Phase Ia/Ib trial of anti-GD2 chimeric monoclonal antibody 14.18 (ch14.18) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) in metastatic melanoma.

We performed a phase Ia/Ib trial of chimeric anti-GD2 monoclonal antibody 14.18 (ch14.18) in combination with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) to determine the maximum tolerated dose as well as immunologic and biologic responses to the regimen. Sixteen patients with metastatic malignant melanoma received escalating doses of ch14.18 (15-60 mg/m2) administered intravenously for 4 h on day 1. Twenty-four hours later, subcutaneous injections of rhGM-CSF were administered daily for a total of 14 days. Significant side effects were related to ch14.18 infusion and consisted of moderate to severe abdominal and/or extremity pain, blood pressure changes, headache, nausea, diarrhea, peripheral nerve dysesthesias, myalgias, and weakness. Dose-limiting toxicity was observed at 60 mg/m2 and consisted of severe hypertension, hypotension, and atrial fibrillation in one patient each, respectively. Significant increases in white blood cell count, granulocyte count, eosinophil count, and monocyte count occurred after rhGM-CSF treatment. Significant enhancement of in vitro and in vivo monocyte and neutrophil tumoricidal activity and antibody-dependent cellular cytotoxicity along with significant elevations in C-reactive protein and neopterin were observed. Despite these immunological and biological changes, no antitumor activity was seen. In short, the combination of ch14.18 and rhGM-CSF resulted in toxicity similar to that observed with ch14.18 alone without improvement in tumor response.

Combination therapy with ifosfamide and liposome-encapsulated muramyl tripeptide: tolerability, toxicity, and immune stimulation.

A phase IIb trial using liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (L-MTP-PE) in combination with ifosfamide (IFX) for patients with relapsed osteosarcoma was undertaken to determine (a) the tolerability of the combination therapy, (b) if L-MTP-PE increased the toxicity of IFX, and (c) whether IFX altered or suppressed the in vivo immune response to L-MTP-PE. Patients had histologically proven osteosarcoma and pulmonary metastases that either developed during adjuvant chemotherapy or were present at diagnosis, persisted despite chemotherapy, and recurred following surgical excision. Stratum A patients were rendered clinically free of disease within 4 weeks of study entry prior to receiving combination therapy. IFX was administered at 1.8 g/m2 for 5 days every 21 days for up to eight cycles. L-MTP-PE was administered twice weekly for 12 weeks, then once weekly for 12 weeks. Once cycle of combination therapy was defined as 5 days of IFX and 3 weeks of L-MTP-PE therapy. Stratum B patients had measurable disease at study entry that was judged to be amenable to surgical resection. Stratum B patients received three cycles of combination therapy prior to surgery to judge clinical and histologic response. Postoperatively, patients received an additional five cycles. A total of nine patients were entered into the protocol: six on stratum A and three on stratum B. Serial blood samples were collected and assayed for cytokine levels (tumor necrosis factor-alpha [TNF alpha], interleukin-6 [IL-6], IL-8, neopterin, C-reactive protein). In addition, peripheral blood monocyte tumoricidal activity was evaluated pre- and post-combination therapy. Complete blood counts with differential and platelet counts were followed weekly. No increase in the toxic side effects of IFX was demonstrated when administered with L-MTP-PE nor were delays in IFX administration due to neutropenia experienced. The toxic side effects of L-MTP-PE were also not increased. Elevations of serum C-reactive protein, plasma neopterin, IL-6, IL-8, and TNF alpha following combination therapy were similar to those observed in patients treated with L-MTP-PE alone. Monocyte-mediated tumoricidal activity was elevated 24 and 72 h following L-MTP-PE and IFX therapy, similar to what has been reported following L-MTP-PE alone. Tumor specimens obtained from stratum B patients showed the histologic characteristics consistent with a "chemotherapy effect," i.e., dead, amorphous, acellular osteoid with cell drop-out.(ABSTRACT TRUNCATED AT 400 WORDS)