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Background The pathophysiological alterations in response to shunt correction in patients with pulmonary arterial hypertension associated with congenital heart disease (CHD-PAH) is still not clear. Purpose To explore the dynamic plasma metabolite profiling and its relationship with clinical characteristics in patients with CHD-PAH during the perioperative period of defect repair. Methods Plasma samples from 13 patients with CHD-PAH were harvested at four time points: before cardiopulmonary bypass (CPB) after anesthesia (Pre), immediately after CPB (T0), 24 h (T24), and 48 h (T48) after defect repair. The untargeted metabolomics strategy based on UPLC Q-TOF MS was used to detect the metabolites. Clinical measures were recorded at indicated time points. Results The sample distribution at four time points was well separated (Figure 1A, B). 193 metabolites were distinguished at different time points according to Variable Important in Projection (VIP) score (Figure 1C), enriched in pathways such as carnitine synthesis, phospholipid biosynthesis and oxidation of branched chain fatty acids (Figure 2A). 17 metabolites alterations were significantly correlated with gradients in mean pulmonary arterial pressure (mPAP) at T48 versus Pre, indicative of the response to defect correction (Figure 2B). Intriguingly, 4 out of the 17 (23.5%) metabolites including propionylcarnitine, butenylcarnitine, isobutyryl-L-carnitine and hexanoylcarnitine were enriched in oxidation of branched chain fatty acids. They were increased at both T24 and T48 (all P Conclusions This is the first study to show the altered metabolic profiles of CHD-PAH patients in perioperative period of defect repair. Metabolites that respond to shunt correction could be a suitable non-invasive marker and would be of great value in disease monitoring and evaluating future therapeutic interventions. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): 13th Five-Year Plan–Precise Medicine–Key Research and Development Program–Clinical Cohort of Rare Disease; National Natural Science Foundation of China Figure 1. Overall analysis of metabolitesFigure 2. Shunt correction associated metabolites

Objective To evaluate the efficacy of a “vacuum sealing drainage (VSD) – artificial dermis implantation (ADI) – thin partial thickness skin grafting (TSG)” sequential therapy for deep and infected wounds in children. Materials and methods Fifty-three pediatric patients with deep and infected wounds were treated with sequential VSD-ADI-TSG therapy. The efficacy of this treatment was compared with that of the surgical debridement-change dressings-thin partial thickness skin grafting previously performed on 20 patients. Survival of tissue grafts, color and flexibility, subcutaneous fullness and scar formation of the graft site were examined and compared. Results The sequential therapy combined the advantages of the VSD treatment, in reducing tissue necrosis and infection on the wound surfaces and promoting the growth of granulation tissue, with the enhancement of grafting by artificial dermis. Compared with the 20 controls, skin grafted on the artificial dermis was more smooth and glossy, while the textures of the region were more elastic, and the scars were significantly lighter in Vancouver scale. Conclusion The sequential VSD-ADI-TSG therapy is a simple and effective treatment for children with deep and infected wounds. Level of evidence IV.

Osteosarcoma is a malignant bone tumor with high incidence. The prognosis of osteosarcoma is very poor when it is diagnosed with metastasis. Numerous studies have demonstrated that aberrant expressions of microRNAs are involved in cancer initiation and development. However, the potential role of miR-214 in osteosarcoma remains largely unrevealed. The current study investigated the relationship between the miR-214 and TNF receptor-associated factor 3 (TRAF3) in osteosarcoma tissues and cell lines. We also aimed to evaluate the potential roles of miR-214 on the occurrence and metastasis in osteosarcoma and verify its effect on the regulation of TRAF3.The miR-214 expression and TRAF3 expression in osteosarcoma tissue samples and cell line were measured using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Followed by transfection assays, transwell assay was conducted to detect the migration and invasion abilities of osteosarcoma cells. Subsequently, Western blotting and luciferase reporter assay were performed in osteosarcoma cells to confirm the target of miR-214.The results showed that miR-214 expression levels were significantly increased not only in osteosarcoma tissues but also in osteosarcoma cell lines as compared with adjacent normal tissues and matched cell lines, respectively. On the contrary, the TRAF3 expression levels in osteosarcoma tissues and cell lines were frequently decreased compared to the control group. Moreover, TRAF3 was identified as a direct target of miR-214 and the inverse relationship between them was also observed in osteosarcoma tissues. Additionally, we found that miR-214 restoration could significantly promote osteosarcoma cell invasion and migration via targeting TRAF3.MicroRNA-214 functioned as an oncogene in osteosarcoma via targeting TRAF3, which may provide new insights into osteosarcoma prevention and treatment.

It is imperative to research the treatment strategy for infections caused by multi-drug resistant (MDR) bacteria, as there are increasing reports showing that more and more patients are decimated by the infections of MDR bacteria and the development of antimicrobial drugs is in downturn. Current researches mainly focus on the following three aspects: developing new antimicrobial agents with the aid of basic scientific achievements in finding new antibacterial targets, achieving antimicrobial purpose by specific lysis of host bacteria with phages of high specificity, and killing bacteria potently by destroying its cytomembrane using broad-spectrum antimicrobial peptides.

The aim of this study was to investigate cytokine messenger RNA (mRNA) expression by peripheral blood mononuclear cells (PBMCs) from renal recipients experiencing acute rejection by blocking OX40-OX40L interactions with recombinant human OX40-Fc fusion protein (rhOX40Fc) in vitro.PBMCs were isolated from 20 recipients experiencing acute rejection episodes (rejection group) and 20 recipients with stable graft function (stable group). Levels of Th1 (interferon [IFN]-γ) and Th2 (interleukin [IL]-4) mRNA expressions by PBMCs were measured using real-time reverse transcriptase-polymerase chain reactions.IFN-γ mRNA expression levels were significantly higher in the rejection than the stable group (P .05). Levels of IL-4 mRNA expression were not significantly different. Among the rejection group, rhOX40Fc reduced significantly the expression of IFN-γ and IL-4 mRNA by anti-CD3-monoclonal antibody stimulated PBMCs (P .05, and P .01, respectively).Blocking of the interaction between OX40 and OX40L in vitro inhibited production of Thl and Th2 type cytokines.

Host defenses against experimental listeriosis in mice involve neutrophils, macrophages, NK cells, and alpha beta T cells. Recently gamma delta T cells have also been implicated in antilisterial resistance. However, their specific role has remained unclear. Here we show that efficient resistance to infection by this bacterium depends on the functions of both alpha beta and gamma delta T cells in both primary and secondary responses. We also present evidence that these functions are complementary. In the livers of alpha beta T cell-depleted mice, bacteria grow to large numbers within hepatocytes but are infrequently found extracellularly. Granulomatous lesions are more frequent and somewhat larger than in normal controls, but remain focal. Neutrophils are absent from liver lesions in these mice. In contrast, the livers of gamma delta T cell-depleted mice contain many extracellular bacteria, but do not show hepatocytes containing large numbers of Listeria. Liver lesions in gamma delta T cell-depleted mice are far more extensive than in normal controls or in alpha beta T cell-depleted mice, and contain large numbers of neutrophils. Particularly in secondary listeriosis, gamma delta T cell-depleted mice show vast coalescent areas of necrotic liver parenchyma within 48 h after infection. Because the bacterial numbers in gamma delta T cell-depleted mice remain lower than in alpha beta T cell-depleted mice, increased mortality in the former may be in part caused by liver failure. We conclude that gamma delta T cells are required to control inflammatory reactivity and to prevent excessive liver damage during the immune response to Listeria monocytogenes.

The juxtaposition of maternal and fetal tissues in the hemochorial placenta has led to speculation that maternal recognition of fetal Ags present on trophoblasts might play an important role in reproductive biology. We report here for the first time such recognition of trophoblasts by T lymphocyte hybridomas representative of certain cells present in the maternal decidua. Trophoblast recognition is TCR dependent and is mediated by members of the V gamma 1+ subset of gamma delta T lymphocytes, a population that we previously have shown to be associated with heat shock protein-60 reactivity. Recognition occurred in experiments in which trophoblast clones or freshly prepared trophoblasts were used and requires cell-cell interaction. Although the maternal-fetal immune relationship typically has been cast in terms of an allograft, the T cell recognition of trophoblasts as described herein is not MHC-restricted, inasmuch as freshly prepared trophoblasts from beta 2-microglobulin-deficient mice were found to be stimulatory. Furthermore, the trophoblast ligand that mediates this recognition is probably a conserved mammalian molecule, because a human trophoblast cell line is also stimulatory. Our findings suggest a novel form of T cell recognition, and may provide an enhanced understanding of the maternal-fetal immune relationship.

Mouse gamma delta T cell hybridomas from various strains that express a TCR-V gamma 1/V delta 6 respond weakly to an autologous Ag and more strongly to a short segment of the mycobacterial heat shock protein-60 (HSP-60). However, V gamma 1/V delta 6 hybridomas derived from AKR mice show greatly reduced or absent responses to these stimuli. To determine whether the lack of response in these AKR hybridomas is caused by polymorphisms found in the expressed AKR gamma and TCR-delta genes or, instead, stems from other genes in AKR, we crossed an AKR mouse with a responder mouse, C57BL/10 (B10), and prepared hybridomas from F1 progeny. Expression of an AKR V gamma 1-J gamma 4-C gamma 4 gene correlated with nonresponsiveness, whereas expression of a B10 V gamma 1-J gamma 4-C gamma 4 gene in most hybridomas ensured responses to both self Ag and the HSP-60 peptide. An allelic difference in the expressed V gamma 6 gene was irrelevant to these responses. Moreover, transfection of a functional B10 V gamma 1-J gamma 4-C gamma 4 gene into an F1 hybridoma variant that had lost the AKR version of this gene restored responses. The allelic gamma gene products differ at nine amino acids in the V region, and three amino acids in the C region. In addition, the AKR C gamma 4 region contains a 16-amino acid insertion. We propose that amino acid differences among those encoded by the AKR V gamma 1-J gamma 4-C gamma 4 gene are responsible for the lack of response, and reduce the ability of the TCR-gamma delta to bind the relevant Ag.

Hybridomas representing the V gamma 1-positive subset of murine gamma delta T cells secrete lymphokines in response to synthetic peptides representing a short segment of the mycobacterial 60-kDa heat shock protein (HSP-60). Here we show the TCR dependency of this response by transfection of productively rearranged TCR genes derived from an HSP-60 reactive gamma delta T cell hybridoma. We also have defined structural requirements for the stimulatory peptide. The smallest HSP-60 peptide capable of stimulating these hybridomas is seven amino acids long, representing positions 181-187, and having the sequence FGLQLEL. Amino acid-substituted derivatives of this peptide, and another containing the same core, p180-190, revealed amino acids essential for stimulatory activity. Phenylalanine in position 181 and leucine in position 183 seem to be required for stimulation of all HSP-60 reactive cells, whereas others are only required by some. Clonal differences in the responses to these peptides provide indirect evidence for cognate TCR-peptide interactions. The smallest stimulatory peptide, p181-187, represents an area not well conserved among HSP-60 molecules of other species, and stimulates a mycobacteria-specific response unlike the earlier observed cross-reactive responses of the same hybridomas with longer HSP-60 peptides derived from mycobacteria and other species (our manuscript in preparation). We propose that the TCR-dependent multiclonal gamma delta T cell response to HSP-60 peptides and derivatives, which in some ways resembles superantigen responses and in other ways resembles responses to conventional Ag, may be a separate, third type of Ag response by T cells.

Murine V gamma 5 and V gamma 6+ T lymphocytes develop sequentially in the thymus during fetal and newborn life, giving rise to intraepithelial lymphocytes of the epidermis and female reproductive tract. In analyzing a panel of gamma delta T cell hybridomas derived from various tissues, we found that all V gamma 6+ cells tested (n = 25) expressed V gamma 5-C gamma 1 mRNA, whereas none of a panel of V gamma 6- cells analyzed (n = 33) expressed this mRNA. V gamma 6 mRNA was rare in V gamma 5+ cells (1 of 9), and absent in all other gamma delta T cells (n = 10). These findings suggest that most, if not all, V gamma 6-TCR+ lymphocytes arise from precursor cells that have previously rearranged the V gamma 5 gene, and indicate that V gamma 5 and V gamma 6+ lymphocytes belong to a common, distinct lineage that does not give rise to other gamma delta T cells.

To evaluate the effect of tissue flap pedicled with the superficial temporal artery in repairing deformity of the head and face in children.From October 1986 to December 1996, 13 children with deformity of the head and face were repaired by this tissue flap. Among them, there were congenital deformity in 9 cases, burned scar in 3 cases and infection scar in 1 case. Among the flaps, 1 was temporal skin flap, 3 were temporal flap with hairbearing scalp, 1 was frontal skin flap, and 8 were posterio-uricular superficial fascia flap and skin flap. The area of tissue flap was ranged from 5.0 cm x 1.2 cm to 10.0 cm x 5.0 cm. The length of the pedicle was 5-8 cm.All tissue flaps healed with first intention. Followed up for 6 months to 12 years, the appearance and function of tissue flaps were satisfactory.The tissue flap pedicled with the superficial temporal artery is suitable to repair many kinds of deformities of the head and face in children. It has the advantages of good blood supply, closely acceptor area, easy operation and satisfactory appearance.

The mode of within-locus gene action in most genomic regions is termed as the major genomic mode, i.e., it is the within-locus allelic effects in most regions of the genome. Determining whether dominance or overdominance is the major genomic mode is important for two long-standing evolutionary genetics issues: 1. How is the genetic variation in most genomic regions maintained? 2. What is the major mechanism for heterosis? Many efforts have been made, but almost all of them suffer some explanational difficulties. Here we propose an alternative inference approach. It is based on the existent theoretical results on the correlation of the recombination rate and the level of neutral variation in different genomic regions. Positive and negative correlation suggest dominance and overdominance, respectively, as the major genomic mode. Zero correlations imply either few selected sites or about equal composition and distribution of dominant and overdominant regions in the genome, depending on the data distribution. This approach not only avoids all the problems associated with earlier approaches, but it is also particularly useful in organisms where controlled breeding is difficult. Well-corroborated data in Drosophila and recently emerging data in mice and humans all suggest dominance as the major genomic mode.

The receptive field (RF) properties of visual neurons extracellularly recorded from the nucleus lentiformis mesencephali (nLM) in pigeons (Columba livia) were quantitatively analyzed using a workstation computer. These cells were actively spontaneous, and direction-and velocity-selective. Using spatial gratings as visual stimuli, these cells could be divided into three groups: uni- (74%), bi- (17%), and omnidirectional (9%) cells in terms of their directionality. On the basis of their velocity selectivity, they could be named slow cells (84%), preferring low velocity (0.1-11 degrees/s), and fast cells (14%), preferring rapid motion (34-67 degrees/s), with one cell (2%) responding maximally to an intermediate velocity of 18 degrees/ s. These two properties were correlated in the way that all unidirectionals were slow cells, omnidirectionals were fast cells, and bidirectionals were either slow or fast cells including the intermediate cell. Using small targets as visual stimuli, it was found that the majority of cells examined had RFs that each consisted of an excitatory RF (ERF) and an inhibitory RF (IRF) that overlapped. The unidirectionals were mainly of this type of RF structure, whereas the omnidirectionals apparently had ERFs alone. The direction preference of ERF was opposite to that of IRF for unidirectional cells tested, whereas they were perpendicular to each other for one bidirectional cell. The overall responses of these cells resulted from interaction between excitation and inhibition induced by directionally different motion. Under certain conditions, visual responses of a particular cells to a small target moving through its ERF were equal in responsive strength to those to whole-field gratings swept over the screen. It was suggested that optokinetic nystagmus produced by wholefield gratings results from population activity of large group(s) of neurons in some optokinetic nuclei, at least one of which is nLM.

To assess the efficacy of Artesunate on falciparum malaria.A randomized controlled study on the treatment of 90 uncomplicated falciparum malaria patients was carried out with 400 mg of artesunate tablet as a total dose over 3 days, 600 mg over 5 days and 800 mg over 7 days.All patients were cured. Fever clearance time (FCT) and parasite clearance time(PCT) among the three groups were similar. Parasite recrudescence rate within 28 days was 39.3% (11/ 28) in 3 day group, 6.9% (2/29) in 5 day group and 3.4% (1/29) in 7 day group (comparing 5 day group with 3 day group, P0.005, comparing 7 day group with 3 day group, P0.005).It indicated that parasite recrudescence rate may be effectively decreased by prolonging treatment courses.

Background: Inflammation and angiogenesis in the tumor microenvironment play a role in human breast cancer. We previously showed that tumors with high M2 macrophages and high microvessel density (MVD) have a shorter overall survival. The antitumor effect of Toll-like receptor 7 (TLR7) has been previously described whereby its activation can alter the tumor microenvironment to adopt an antitumorigenic (Th-1) immune response and inhibit angiogenesis, with a potential immune mediated benefit in breast cancer. To our knowledge, there are no studies that evaluated TLR7 and its relationship to macrophages and angiogenesis in human breast cancer. Methods: Breast tumors were obtained from the University of Chicago Breast Cancer SPORE tissue bank under IRB approved protocols. Tissue microarrays were constructed and molecular subtype was assigned based on immunohistochemical (IHC) staining into the following groups: luminal A (ER+, PR+, HER2-), luminal B (ER+, HER2+ or ER+, PR-), HER2-like (ER-, HER2+) and basal-like (ER-, HER2-, EGFR+ and/or CK5/6+). Macrophage density was determined using double staining with CD68/CD163. Microvessel density (MVD) was measured by IHC staining using anti-CD34. Staining quantification was performed by a trained pathologist and scoring performed independently by two pathologists. Based on median distribution, we assigned M2 macrophage content and MVD into high vs. low, and the TLR7 expression into faint vs. strong. To evaluate the association between M2 macrophages and TLR7, Spearman's rho correlation coefficients were calculated. Survival analysis was done using Tarone ware statistics. Results: After removal of duplicate cores, we had 100 tumors available for scoring. There was an equal distribution of M2 macrophage content, MVD, and TLR7 expression (p = 0.074, p = 0.908, p = 0.145 respectively). The tumor subtypes were 53.2% luminal A, 34.0% basal-like, 7.4%, luminal B, and 5.3% Her2-like. We found a significant correlation between high number of M2 macrophages and high MVD (spearman's rho correlation coefficient = 0.343, p = 0.004). There was an association between M2 macrophages and TLR7 expression (Pearson χ2 = 4.425, p = 0.025). Compared to tumors with faint TLR7 expression, those with strong TLR7 expression exhibited a longer overall survival (Tarone ware statistic 5.053, p = 0.025) and this continued to be significant when adjusted for M2 macrophage content (Tarone ware statistic = 4.049, p = 0.044) and for MVD (Tarone ware statistic = 4.042, p = 0.044). There was no statistically significant association between TLR7 expression and molecular subtypes of breast cancer. Conclusion: Tumor associated macrophages (M2 macrophages) and angiogenesis (high MVD) are known to contribute to worse outcomes in human breast cancer. Our results suggest that activation of TLR7 can have a protective effect in human breast cancer even in tumors harboring features of more aggressive disease. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-01-04.

Acquired inhibitors of factor V are rare causes of clinical bleeding, whose severity ranges from mild to life-threatening. Optimal treatment of patients with factor V inhibitors is uncertain. We report on our successful treatment approach in a patient with spontaneous, life-threatening intracranial bleeding caused by a factor V inhibitor. The patient deteriorated after initial treatment with fresh-frozen plasma and platelet transfusions. He was subsequently treated with a combination of plasma exchange and chemotherapy, which led to complete recovery. Our experience suggests that plasma exchange may be life-saving in cases of severe bleeding caused by factor V inhibitors. The use of plasmapheresis in conjunction with chemotherapy is an efficacious and well-tolerated treatment and should be considered in patients with factor V inhibitors.

Twenty-seven patients with gametocytes of Plasmodium falciparum (PF) were divided into groups A, B, and C. A daily dose of 1200 mg artemisinin was given for 5 days to group A, a state dose of 750 mg of mefloquine to group B and a single dose of 750 mg mefloquine combined with 45 mg primaquine to group C. After treatment, the gametocyte count was taken daily, and infectivity of the gametocytes to Anopheles dirus via membrane feeding was also studied. Results showed that in group A, the density of gametocyte and infectivity were significantly reduced on days 4, 7, 14 and 21 after treatment; In group B, the gametocytes were significantly reduced on days 7, 14 and 21 and infectivity was significantly cut down on days 14 and 21 after medication. In group C, gametocytes disappeared in 5 out of 9 patients with failure of infecting mosquitoes in all 9 patients on day 4 after treatment. These indicate that artemisinin can effectively influence the infectivity of gametocytes of PF. Artemisinin is much better in blocking the transmission of PF malaria than mefloquine.

Diagnosis and treatment of 95 patients with chronic subdural hematoma (CSDH) are presented. There were 92 males, 56 patients (59%) over 51 years old and 66 of them with history of head injury. The majority of our patients appeared increased intracranial pressure. Mental confusion and incontinence of urine and stool usually occurred in senile patients. Ultrasonic and electroencephalography examination were helpful in location of the hematoma. Cerebral angiography could make actual diagnosis. CT scan were more useful in diagnoses of CSDH and may discover multiple hematomas. If CSDH image were iso-dense on CT scan cerebral angiography should be performed. In the early years extirpation of CSDH including the capsule was performed. Now an irrigation drainages by cranial bur hole was performed usually. 5 patients had complication. The mechanism and prevention of CSDH are discussed.

The DNA of an organism can be digested into smaller fragments, stored individually as clones in phage, for example, to create a clone library, and retrieved later, when needed. The original ordering of fragments is lost in the process of creating the library. Hence, it is important to be able to place clones in order according to their position along chromosome(s), and this process is referred to as "in vitro reconstruction" or "contig mapping" of an organismal genome. Clones in the phage library can be assigned binary call numbers by scoring each clone for hybridization (0 or 1) with a battery of short manufactured DNA sequences called synthetic oligonucleotides or with restriction enzyme digests of each clone. Those clones with similar call numbers are placed close together in the ordered library. We address the design question of how many clones and probes to use to carry out in vitro reconstruction of an organism's chromosomes. This physical mapping problem is placed in the context of coverage problems in geometrical probability. Various statistics are developed to summarize how an ordered library covers a chromosome, the extent of clone overlap, and the similarity between clone call numbers. Several tests for whether clones overlap are given, together with their power properties. A simulation study is used to determine how robust some of the tests for clone overlap are to model violations. Tables are presented for researchers to choose the number of clones and probes on the basis of both power and technical considerations surrounding the hybridization experiments.

Peritoneal elicited macrophages (PEM) from mammary tumor-bearing mice have a decreased capacity to become cytotoxic against syngeneic, allogeneic, and xenogeneic target cells upon in vitro stimulation with LPS, as compared with PEM of normal mice. A regulatory mechanism other than PG release is suggested because the addition of both indomethacin and LPS to macrophage cultures from tumor-bearing mice caused no changes in their cytotoxic capability. Because tumor products have been implicated in the down-regulation of immune responses, we investigated whether pretreatment with supernatants from the tumor cell line DA-3, derived from the in vivo mammary adenocarcinoma D1-DMBA-3, affects the cytolytic capacity of macrophages. This treatment inhibits, in a dose-dependent fashion, the ability of stimulated normal PEM to kill target cells. Partial purification of DA-3 cell line supernatant showed that most of the inhibitory activity was exerted by factors with a molecular mass greater than 10 kDa and less than 30 kDa. However, slight inhibition could also be observed with fractions containing molecules less than 10 kDa. The data suggest that more than one factor released by the mammary tumor cells may be involved in the down-regulation of macrophage-mediated cytotoxicity. Because the DA-3 cells constitutively produce granulocyte-macrophage CSF (GM-CSF), which has a molecular mass of 27 kDa, we pretreated PEM from normal mice in vitro with rGM-CSF for 24 h. This resulted in a dose-dependent decrease in their capacity to kill tumor target cells upon LPS stimulation. Furthermore, PEM from normal mice injected with rGM-CSF for 25 days displayed a profound decrease in their cytolytic ability against DA-3 targets upon in vitro stimulation with increasing amounts of LPS. The pretreatment of PEM from normal mice with a combination of DA-3 cell supernatants and specific anti-GM-CSF partially neutralized the inhibitory effect of the DA-3 supernatant on macrophage tumoricidal capability. These results indicate that tumor-derived GM-CSF is an important factor involved in the decreased macrophage cytotoxicity during mammary adenocarcinoma progression.

Using a dimethylbenzanthracene-induced immunogenic nonmetastatic murine mammary adenocarcinoma in BALB/c mice, our previous work has shown that splenocytes from tumor bearers have reduced responses to both mitogens and Ag including tumor-associated Ag. NK and cytotoxic T cell activities are also reduced in splenocytes of tumor bearers. Mac-1+2+ macrophages induced in mammary tumor bearers are capable of down-regulating lymphocyte responses to mitogens and tumor-associated Ag by cell to cell contact interaction and increased PGE2 production. We have found that the tumor constitutively releases a granulocyte-macrophage (GM)-CSF-like factor in vivo and in vitro, which may be responsible for the systemic increase in cells of the macrophage lineage in tumor-bearing mice. A tumor cell line established from the in vivo tumor expresses and releases GM-CSF as shown by Northern and Western blot analyses. Daily i.p. injections for 3 wk of 10,000 U of rGM-CSF into normal mice induces hemopoietic and immunologic alterations similar to those observed in tumor bearers. Mac-1+ and/or Mac-2+ macrophages can also be detected in the spleens and bone marrow of the mice treated with rGM-CSF. Additionally, splenocytes from rGM-CSF-treated mice have reduced responses to mitogens and their peritoneal exudate cells can cause in vitro down-regulation of proliferative responses of lymphocytes from normal mice. The suppression can be partially reversed by the addition of indomethacin to the cultures suggesting that PGE2 may contribute to the effect. rGM-CSF enhances the in vitro release of PGE2 by the spleen, bone marrow, and peritoneal cells of normal mice. These data indicate that the high levels of GM-CSF constitutively produced by the tumor may be responsible for the hemopoietic changes and immunologic alterations observed in tumor-bearing mice.

Forty patients with a history of recurrent attacks of asthma after ingesting certain foods are described. All of these individuals had positive intracutaneous skin tests to at least one of the food allergens which was suggested by their history to cause an attack. In four of these patients, a bronchoprovocational challenge was performed with an aerosolized extract of the potential offending food. Clinical manifestations, possible mechanisms, and diagnostic measures for food-induced asthma are discussed.

Using an immunogenic nonmetastatic murine mammary adenocarcinoma (D1-DMBA-3) induced in BALB/c mice by dimethylbenzanthracene, we have previously shown that splenocytes from tumor bearers have depressed lymphocyte responses to mitogens and antigens, including tumor-associated antigens. In addition, they display decreased natural killer and T-cell cytotoxic activities. Macrophages from tumor-bearing mice appear to be responsible for the suppression of T- and B-cell responses to concanavalin A, lipopolysaccharide, and tumor-associated antigens observed in tumor bearers. The appearance of these macrophages in the spleen tightly parallels the progressive growth of the tumor and the concomitant immunosuppression. Simultaneously high levels of macrophage progenitors were observed in blood, bone marrow, lung, and liver. A significant increase of colony-stimulating activity of the granulocyte-macrophage lineage was detected in the sera from tumor-bearing mice. Higher levels of this colony-stimulating activity (CSA) were detected in tumor cystic fluid as compared with the levels in serum. A tumor cell line established in vitro from the D1-DMBA-3 in vivo tumor produces high levels of a factor with CSA in culture supernatant fluids. Partial purification of the CSA from the tumor cell line supernatants was achieved using CentriCell ultrafiltration and SephacrylS-300 chromatography. These studies revealed that the molecular weight of the colony-stimulating-like factor is 32,000 to 35,000. The morphology of the colonies obtained in cultures using this factor is similar to that of the colonies that develop in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) but not with macrophage colony-stimulating factor (M-CSF). CSA from tumor cell supernatants was neutralized by antiserum to GM-CSF but not with anti-M-CSF or anti-granulocyte colony-stimulating factor (G-CSF). Macrophages from bone marrow or peritoneal exudates from normal mice cultured with tumor supernatant for 2 to 3 days strongly inhibit normal splenocyte responses to concanavalin A and lipopolysaccharide. The data suggest that the tumor releases a GM-CSF that alters the hemopoietic system and induces or expands macrophages, which exert a suppressive function on the immune system of tumor-bearing mice.

Eighteen asymptomatic Artemisia pollen asthma patients with normal pulmonary function were selected for a double-blind trial of oral immunotherapy. Each patient had a positive skin test to Artemisia pollen extract and also a positive bronchial challenge response to the same extract. The patients were randomly assigned to active treatment or placebo group and received intensive oral administration of Artemisia pollen extract over a 50-day course. The nine patients who received the active treatment ingested a cumulative dose of 396,652 PNU and showed a significant decrease in serum-specific IgE antibodies (P less than .05) and a significant reduction in bronchial sensitivity to the same extract (P less than .01). The changes in these two variables correlated well (r = .8787, P less than .01). The nine patients who received the placebo showed no significant changes in serum-specific IgE or bronchial sensitivity to Artemisia pollen extract. Follow-up of two cases with the same extract showed that the reductions in serum-specific IgE as well as bronchial sensitivity induced by oral immunotherapy were maintained for 3 months.

The DNA of an organism can be digested into smaller fragments, stored individually as clones in phage, for example, to create a clone library, and retrieved later, when needed. The original ordering of fragments is lost in the process of creating the library. Hence, it is important to be able to place clones in order according to their position along chromosome(s), and this process is referred to as "in vitro reconstruction" or "contig mapping" of an organismal genome. Clones in the phage library can be assigned binary call numbers by scoring each clone for hybridization (0 or 1) with a battery of short manufactured DNA sequences called synthetic oligonucleotides or with restriction enzyme digests of each clone. Those clones with similar call numbers are placed close together in the ordered library. We address the design question of how many clones and probes to use to carry out in vitro reconstruction of an organism's chromosomes. This physical mapping problem is placed in the context of coverage problems in geometrical probability. Various statistics are developed to summarize how an ordered library covers a chromosome, the extent of clone overlap, and the similarity between clone call numbers. Several tests for whether clones overlap are given, together with their power properties. A simulation study is used to determine how robust some of the tests for clone overlap are to model violations. Tables are presented for researchers to choose the number of clones and probes on the basis of both power and technical considerations surrounding the hybridization experiments.

Sodium crompromate (SCP) is an isomer of disodium cromoglycate (DSCG). The two chemicals are pharmacologically similar but with different chemical synthesis. A comparative double-blind latin square study of the preventive effect of SCP and DSCG versus a placebo aerosol was carried out on a group of nine cases of exercise-induced asthma (EIA). Compared with the placebo, both SCP and DSCG were able to prevent asthma under exercise provocation, as monitored by chest auscultation and FEV1. There was no significant difference in the percentage fall of FEV1 on challenge. This suggests that SCP has a similar prophylactic effect as that of DSCG on EIA.

The hematoporphyrin derivative YHPD, a China-made product, has been clinically used in photodynamic therapy of tumors as a good photosensitizing drug. The NMR study on the structure of its major components is reported here. In terms of high performance liquid chromatography (HPLC) four major components A, B, C and D were isolated. The NMR results showed that the component A is O-acetylhematoporphyrin, B and C are two isomers of vinyldeuteroporphyrin. The spectra of 2-dimensional homonuclear correlation NMR, 2-dimensional NOE (nuclear overhauser enhancement), 13C-NMR and off-resonance as well as FAB (fast atom bombarding) mass spectrum of component D indicate that it is a protoporphyrin dimer linked by carbon-carbon bond. This finding may provide a chemical basis for understanding the difference in biological activity between YHPD and other foreign commercial HPD, as well as the composition of clinically used alkali-treated HPD and its effective component.

The thymic compartment of mammary tumor bearing mice is severely affected with increasing tumor growth. T cell functions in peripheral organs are greatly impaired or absent in such animals. A study of the thymus in relation to tumorigenesis revealed a profound decrease in size accompanied by a disrupture of the normal thymic architecture. Although a strong splenic suppressor cell activity can be detected in this animal model, splenectomy did not prevent thymic involution. Injection of tumor associated factors into normal mice resulted in a thymic atrophy similar to that seen in tumor bearing mice. These findings indicate that the observed thymic involution may not be only due to stress-related phenomena but also a direct effect of the tumor.