Changes in serum inflammatory factors, adiponectin, intestinal flora and immunity in patients with non-small cell lung cancer.

OBJECTIVE: The aim of this study was to explore the changes in the body state of patients with non-small cell lung cancer (NSCLC), including intestinal flora, serum inflammatory factors, immunity and adiponectin. PATIENTS AND METHODS: A total of 18 NSCLC patients (disease group) and 16 healthy people from the Medical Center (control group) were selected as research objects. The levels of immune molecules immunoglobulin A (IgA), IgG and IgM, and inflammatory factors interleukin-2 (IL-2), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and IL-6 were detected via enzyme-linked immunosorbent assay (ELISA). The level of adiponectin was determined using the quantitative kit. In addition, the changes in intestinal flora were analyzed. RESULTS: The overall survival time of NSCLC patients was significantly affected by IL-2 (p=0.0026), CRP (p=0.03), TNF-α (p=0.014) and IL-6 (p=0.00018). It can be seen that these inflammatory factors may play important roles in the progression of NSCLC. The levels of TNF-α (p=0.037), IL-2 (p=0.043) and CRP (p=0.000) in the peripheral blood serum were significantly higher in disease group than control group. Meanwhile, the levels of IgA (p=0.040) and IgG (p=0.000) in the peripheral blood serum were significantly higher in disease group than control group. However, no significant difference was observed in the level of IgM between the two groups (p>0.05). The expression of adiponectin gene (ADIPOQ) could remarkably affect the overall survival rate of NSCLC patients, and patients with high expression of ADIPOQ exerted significantly better prognosis (p=0.017). The level of serum adiponectin was evidently higher in control group than that in disease group (p<0.05). According to the linear discriminant analysis (LDA) score of the intestinal flora in both groups, the abundance of some intestinal flora (Enterobacter and Lachnospiraceae) was markedly higher in disease group than control group (p<0.05). However, the abundance of Bifidobacteria, Pediococcus and Lactobacillus was remarkably higher in control group than disease group (p<0.05). Correlation analysis indicated that Lactobacillus was positively correlated with Bifidobacteria (r=0.44, p=0.000), whereas was negatively correlated with Enterobacter (r=-0.22, p=0.024). Furthermore, Enterobacter was negatively associated with Bifidobacteria (r=-0.15, p=0.038) and Streptococcus (r=-0.12, p=0.046). CONCLUSIONS: Serum inflammatory factors, adiponectin, intestinal flora and immunity may play important roles in the development of NSCLC.

Correlations of IL-6 and IL-10 gene polymorphisms with childhood acute lymphoblastic leukemia.

OBJECTIVE: The aim of this study was to investigate the correlations between interleukin-6 (IL-6) and IL-10 gene polymorphisms with childhood acute lymphoblastic leukemia. PATIENTS AND METHODS: Specimens were collected from 200 children with acute lymphoblastic leukemia (disease group) and 200 normal children (control group) in our hospital. DNA was extracted from peripheral blood nucleated cells in both groups to detect the gene polymorphisms rs2069830 and rs2069836 of IL-6, as well as rs3024489 and rs3024493 of IL-10. Then, the content of serum IL-6 and IL-10 was determined via enzyme-linked immunosorbent assay (ELISA). RESULTS: It was found that there were differences in the distribution of alleles of IL-6 gene polymorphism rs2069830 (p=0.000) and IL-10 gene polymorphism rs3024493 (p=0.007) between the disease group and control group. The frequency of T allele of IL-6 gene polymorphism rs2069830 was higher, while that of IL-10 gene polymorphism rs3024493 was lower in the disease group. Besides, the differences in the distribution of genotypes of IL-6 gene polymorphism rs2069830 (p=0.000) and IL-10 gene polymorphism rs3024493 (p=0.000) were also observed between the disease group and control group. Moreover, the disease group had higher frequencies of TT genotype of IL-6 gene polymorphism rs2069830 and TA genotype of IL-10 gene polymorphism rs3024493. The frequencies of dominant model of IL-6 gene polymorphism rs2069830 (p=0.048) and recessive model of IL-10 gene polymorphism rs3024493 (p=0.000) in the disease group were different from those in the control group. In addition, the frequency of CC + CT dominant model of IL-6 gene polymorphism rs2069830 was lower, and the frequency of TA + AA recessive model of IL-10 gene polymorphism rs3024493 was higher in the disease group. There were differences in haplotypes CG (p=0.001), CT (p=0.007), and TG (p=0.000) of IL-6 gene, as well as haplotypes AA (p=0.002) and AT (p=0.005) of IL-10 gene between disease group and control group. Furthermore, the content of IL-6 in the serum was associated with the genotypes of IL-6 gene polymorphism rs2069830 (p<0.05), whereas the children with acute lymphoblastic leukemia carrying CT genotype had remarkably higher content of serum IL-6. The genotypes of IL-6 gene polymorphism rs2069830 was notably related to white blood cell (WBC) (p=0.002), and the WBC level was higher in children with CT genotype. The genotypes of IL-10 gene polymorphism rs3024489 had prominent correlations with platelet (PLT) (p=0.043), and the children with AA genotype had a higher PLT level. In addition, the genotypes of IL-10 gene polymorphism rs3024493 were evidently correlated with hemoglobin, which was significantly higher in children carrying TA genotype. CONCLUSIONS: The gene polymorphisms of IL-6 and IL-10 are significantly correlated with the susceptibility to and pathogenesis of childhood acute lymphoblastic leukemia.

A high-mobility two-dimensional electron gas at the spinel/perovskite interface of γ-Al2O3/SrTiO3.

The discovery of two-dimensional electron gases at the heterointerface between two insulating perovskite-type oxides, such as LaAlO(3) and SrTiO(3), provides opportunities for a new generation of all-oxide electronic devices. Key challenges remain for achieving interfacial electron mobilities much beyond the current value of approximately 1,000 cm(2) V(-1) s(-1) (at low temperatures). Here we create a new type of two-dimensional electron gas at the heterointerface between SrTiO(3) and a spinel γ-Al(2)O(3) epitaxial film with compatible oxygen ions sublattices. Electron mobilities more than one order of magnitude higher than those of hitherto-investigated perovskite-type interfaces are obtained. The spinel/perovskite two-dimensional electron gas, where the two-dimensional conduction character is revealed by quantum magnetoresistance oscillations, is found to result from interface-stabilized oxygen vacancies confined within a layer of 0.9 nm in proximity to the interface. Our findings pave the way for studies of mesoscopic physics with complex oxides and design of high-mobility all-oxide electronic devices.

Expression of beclin 1, an autophagy-related protein, in human cervical carcinoma and its clinical significance.

PURPOSE: To investigate the impact of beclin 1 on prognosis of cervical cancer, we determined the expression of beclin 1 in cervical cancer, cervical intraepithelial neoplasia (CIN) and normal cervical tissues. METHODS: A total of 122 cases of cervical cancer, 35 cases with CIN and 31 cases with uterine fibroids were collected at the Cancer Center of Sun Yat University to determine the expression of beclin 1. RESULTS: Beclin 1 positive rate in normal cervical tissues, CIN tissues and cervical cancers was 83.9%, 74.3% and 53.3%, respectively, and it was significantly different between the three groups (p < 0.01). Beclin 1 expression was negatively correlated with cervical cancer differentiation, lymph node metastasis, recurrence and death (p < 0.05). The negative expression is the risk factor affecting overall survival (p < 0.05) and progression-free survival (PFS) (p < 0.05). Multivariate analysis showed that beclin 1 negative expression was an independent risk factor of PFS time. CONCLUSIONS: Beclin 1 may play a role in the occurrence and development of cervical cancer. Beclin 1 positive expression in patients indicates a better prognosis.

Prediction model of pelvic lymph node metastasis in early stage cervical cancer and its clinical value.

AIM: This study was designed to investigate the risk factors of pelvic lymph node metastasis in early stage cervical cancer in order to establish a prediction model for this metastasis and to explore the feasibility of conservative surgery. METHODS: The records of 207 stage IB-IIA cervical cancer patients were retrospectivly analyzed. The risk factors of pelvic lymph node metastasis were analyzed using univariate and multivariate methods. The prediction model for pelvic lymph node metastasis was established by logistic regression. RESULTS: Without preoperative adjuvant therapy, the metastatic rate of pelvic lymph node in stage IB-IIA cervical cancer was 25.1%. The serum SCCAg, the tumor diameter, the depth of cervical stroma invasion, and the cervical canal involvement were revealed as the risk factors of pelvic lymph node metastasis by univariate analysis (P<0.05). Multivariate analysis showed that the serum SCCAg and the depth of cervical stroma invasion were the independent risk factors of pelvic lymph node metastasis (P<0.05, OR = 6.917, 2.227). The patients were divided into three groups according to different independent risk factors: the low-risk group, the medium-risk group, and the high-risk group, which showed metastatic rates of pelvic lymph node of 5.7%, 16.9%, and 48.7%, respectively (P<0.001). A prediction model for pelvic lymph node metastasis was established as follows: Logti(P) = -2.534 + serum SCCAg×1.934 + depth of cervical stroma invasion×0.801. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of this prediction model were 53.8%, 83.9 %, 52.8%, 84.4%, and 76.3%, respectively. CONCLUSION: The serum SCCAg and the depth of cervical stroma invasion were the independent risk factors of pelvic lymph node metastasis in early stage cervical cancer. The proposed prediction model may help to improve the conservative surgery for early stage cervical cancer.

Molecular pathways that modify tumor radiation response.

Aberrant expression of signal transduction molecules in pathways controlling cell survival, proliferation, death, or differentiation are a common feature of all tumors. The identification of the molecules that are involved allows the development of novel tumor-specific strategies. Not surprisingly, targeting these pathways often also results in radiosensitization. The efficacy of such directed therapies may, however, be limited by the heterogeneity and the multiple mutations that are associated with the cancerous state. A more robust alternative may be to target global mechanisms of cellular control. The ubiquitin/proteasome degradation pathway is one candidate for such therapeutic intervention. This pathway is the main posttranscriptional mechanism that controls levels of many short-lived proteins involved in regulation of cell cycle progression, DNA transcription, DNA repair, and apoptosis. Many of these proteins are involved in various malignancies and/or radiation responses. In recent years, proteasome inhibitors have gained interest as a promising new group of antitumor drugs. PS-341, a reversible inhibitor of proteasome chymotryptic activity, is currently being tested in phase I clinical trials. In this study, we show that proteasome inhibition by PS-341 can alter cellular radiosensitivity in vitro and in vivo, in addition to having direct antitumor effects.

[Altered behaviour and expression of Fos in rats born in hypergravity and their re-adaptation to the normal gravity].

Changes in behaviour relevant to the vestibular system were studied in Long-Evans rats which were fertilized, born and housed in 2 acceleration of gravity for 4 months and thereafter exposed to 1 acceleration of gravity, and expression of Fos protein in the brain stem was examined. Data from the hypergravity rats were compared respectively with those from the rotation group and the labyrinthectomized group. Static and locomotion modes of the hypergravity rats were changed, tension of extensor was enhanced and the abilities in locomotion equalization and orientation in swimming and air-righting response were reduced. The adaptation process varied with different behaviours. The time for recovery of the ability of orientating in swimming was the longest, taking more than 1 month. The Fos protein expression provides a useful tool for mapping brain functional activities after sensory stimulation, showing a low basal level in normal and labyrinthectomized groups. The hypergravity rats, on the other hand, exhibited more Fos-positive cells in the superior colliculus, inferior colliculus, periaqueductal gray, raphe dorsal nucleus and solitary nucleus. In contrast, the inferior olivary nuclei, locus coeruleus and vestibular nuclei were not strongly labeled. These spatial patterns of Fos expression suggest that a decrease in gravity-inertial force may activate a neural pathway different from the vestibulo-olivar pathways activated by an increase in gravity-inertial force.

Induction of c-fos and junB mRNA following in vivo brain irradiation.

Although radiotherapy is a front line treatment for brain tumors, little is known about the in vivo molecular responses of brain to irradiation. In this study, expression of c-fos, c-jun and junB immediate-early genes were followed in mouse brain after irradiation. C-fos and junB, but not c-jun, mRNA was induced within 15 min in unanesthetized irradiated mice. Induction was transient and lasted < 4 h. The response was dose-dependent with increases in c-fos and junB mRNA levels after dose of > or = 2 and 7 Gy, respectively. Anesthesia of mice with pentobarbitol delayed the increases in mRNA expression and the response was attenuated. Pre-treatment of mice with dexamethasone, in a schedule which suppressed acute-phase gene expression after brain irradiation, did not significantly change c-fos and junB induction. Our results show that c-fos and junB responses occur in the brain in response to irradiation and that they can be modified by pentobarbital treatment but suggest that there is no direct correlation between the level of mRNA expression and later expression of cytokines or other acute-phase response genes.

Delayed molecular responses to brain irradiation.

The chance of life-threatening complications occurring late after brain irradiation limits the efficacy of this form of cancer therapy. The molecular and cellular events that trigger radiation-induced brain damage are still unknown, but since they have the potential to serve as valuable targets for therapeutic intervention they are worth delineating. In this murine study, the effect of irradiation on the expression of molecules which are known to contribute to brain damage in other model systems was examined. Expression of genes encoding cytokines (TNF-alpha/beta, IL-1 alpha/beta, IL-2, IL-3, IL-4, IL-5, IL-6 and IFN-gamma), cytokine receptors (TNF-Rp55 and p75, IL-1R- p60 and p80, IFN-gamma R, and IL-6R), the cell adhesion molecule (ICAM-1), inducible nitric oxide synthetase (iNOS), anti-chymotrypsin (EB22/5.3), and the gliotic marker (GFAP) was evaluated over a 6-month period using a sensitive RNase protection assay (RPA). We had previously demonstrated that within 24 h of brain irradiation there is an acute transitory molecular response involving TNF-alpha, IL-1, ICAM-1, EB22/5.3 and GFAP. This study shows re-elevation of TNF-alpha, EB22/5.3 and GFAP mRNA levels at 2-3 months, but only TNF-alpha mRNA was overexpressed at 6 months. These time points are when neurological abnormalities are seen after higher doses. The data suggest that TNF-alpha may be involved in late brain responses to irradiation and could contribute to clinical symptoms.

Induction of acute phase gene expression by brain irradiation.

PURPOSE: To investigate the in vivo acute phase molecular response of the brain to ionizing radiation. METHODS AND MATERIALS: C3Hf/Sed/Kam mice were given midbrain or whole-body irradiation. Cerebral expression of interleukins (IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6), interferon (IFN-gamma), tumor necrosis factors (TNF-alpha and TNF-beta), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthetase (iNOS), von Willebrand factor (vWF), alpha 1-antichymotrypsin (EB22/5.3), and glial fibrillary acidic protein (GFAP) was measured at various times after various radiation doses by ribonuclease (RNase) protection assay. The effects of dexamethasone or pentoxifylline treatment of mice on radiation-induced gene expression were also examined. RESULTS: Levels of TNF-alpha, IL-1 beta, ICAM-1, EB22/5.3 and to a lesser extent IL-1 alpha and GFAP, messenger RNA were increased in the brain after irradiation, whether the dose was delivered to the whole body or only to the midbrain. Responses were radiation dose dependent, but were not found below 7 Gy; the exception being ICAM-1, which was increased by doses as low as 2 Gy. Most responses were rapid, peaking within 4-8 h, but antichymotrypsin and GFAP responses were delayed and still elevated at 24 h, by which time the others had subsided. Pretreatment of mice with dexamethasone or pentoxifylline suppressed radiation-induced gene expression, either partially or completely. Dexamethasone was more inhibitory than pentoxifylline at the doses chosen. CONCLUSIONS: The initial response of the brain to irradiation involves expression of inflammatory gene products, which are probably responsible for clinically observed early symptoms of brain radiotherapy. This mechanism explains the beneficial effects of the clinical use of steroids in such circumstances.

Lack of differential radiosensitization of hypoxic cells in a mouse tumor at low radiation doses per fraction by cisplatin.

Korbelik and Skov (Radiat. Res. 119, 145-156, 1989) have reported that cis-diamminedichloroplatinum (II) (cisplatin) shows substantial preferential radiosensitization of hypoxic cells in vitro at low radiation doses (1-4 Gy), and that the interaction seen with low doses of radiation is greatly diminished at high radiation doses. If such an interaction occurred with fractionated irradiation in vivo, it would be extremely important to radiation therapy, since the sensitizer enhancement ratios achievable in the low-dose region are higher than those achievable with current hypoxic cell radiosensitizers. We have tested this possibility in an experimental mouse tumor using fractionated irradiation under conditions in which the response of the tumor was determined by either its aerobic or its hypoxic cells. RIF-1 tumors were irradiated with 10 fractions of 1-4 Gy every 12 h with cisplatin given either as 12 mg/kg once before the first radiation dose or as 1.2 mg/kg at various times prior to each radiation dose. The tumors were irradiated with or without a clamp applied 2-3 min before each radiation dose. The effectiveness of the treatments was assayed by regrowth delay. Cisplatin caused a similar regrowth delay when used alone in both clamped and nonclamped tumors and produced a similar additive or supra-additive interaction when used with the 10 fractionated radiation schedule whether the tumors were hypoxic or aerobic. Our data suggest that cisplatin does not show any preferential radiosensitization of hypoxic cells with low-dose multifraction irradiation in this tumor, although a clear schedule-dependent interaction between the drug and radiation was seen for both aerobic and hypoxic tumors.

scid mutation in mice confers hypersensitivity to ionizing radiation and a deficiency in DNA double-strand break repair.

C.B-17 severe combined immunodeficient (scid) mice carry the scid mutation and are severely deficient in both T cell- and B cell-mediated immunity, apparently as a result of defective V(D)J joining of the immunoglobulin and T-cell receptor gene elements. In the present studies, we have defined the tissue, cellular, and molecular basis of another characteristic of these mice: their hypersensitivity to ionizing radiation. Bone marrow stem cells, intestinal crypt cells, and epithelial skin cells from scid mice are 2- to 3-fold more sensitive when irradiated in situ than are congenic BALB/c or C.B-17 controls. Two independently isolated embryo fibroblastic scid mouse cell lines display similar hypersensitivities to gamma-rays. In addition, these cell lines are sensitive to cell killing by bleomycin, which also produces DNA strand breaks, but not by the DNA crosslinking agent mitomycin C or UV irradiation. Measurement of the rejoining of gamma-ray-induced DNA double-strand breaks by pulsed-field gel electrophoresis indicates that these animals are defective in this repair system. This suggests that the gamma-ray sensitivity of the scid mouse fibroblasts could be the result of reduced repair of DNA double-strand breaks. Therefore, a common factor may participate in both the repair of DNA double-strand breaks as well as V(D)J rejoining during lymphocyte development. This murine autosomal recessive mutation should prove extremely useful in fundamental studies of radiation-induced DNA damage and repair.

Enhancement of the antitumor effect of flavone acetic acid by the bioreductive cytotoxic drug SR 4233 in a murine carcinoma.

Flavone acetic acid (FAA, NSC 347512) is a new anticancer drug currently undergoing clinical investigation. Although the precise mechanism for its broad spectrum of activity against transplanted murine solid tumors is unknown, it has been reported that FAA reduces tumor blood flow and produces hemorrhagic necrosis. We have confirmed this finding with the murine transplanted carcinoma SCCVII: 200 mg/kg FAA reduced tumor blood flow to 20-30% of normal for 1-2 days as determined by rubidium 86 extraction. In an attempt to exploit the tumor hypoxia produced by FAA, we have combined it with the novel bioreductive drug SR 4233, a benzotriazine dioxide with high selective toxicity for hypoxic cells. Marked enhancement of the antitumor effect of FAA (200 mg/kg) was observed when it was combined with SR 4233 (0.1 and 0.2 mmol/kg). This was seen using tumor cell survival, regrowth delay, and histological endpoints, with the best results obtained when the two agents were injected simultaneously. These data suggest that targeting bioreductive cytotoxic agents to tumors by producing tumor hypoxia may be a valid way of increasing the tumor cell killing of these agents.

Establishment, characteristics, and utilization of a new in vivo-in vitro system.

Radiotherapy and chemotherapy are two important methods for malignant tumor treatment. To research radiobiological response in therapy, we have established a better experimental method in contrast to the traditional ones such as TCD50, regrowth delay, cell survival curve, etc, all with their limitations. A new mouse tumor in vivo-in vitro system LA795 Vv-Vt has been developed for studies on radiobiology. Such a system could be used to study the in vivo response of a solid tumor by the in vitro cloning assay. For the purpose of increasing the PE in vitro, LA795 Vv-Vt tumor line was purified through culturing the cells as a clonogenic spheroid. The spheroids were then injected into the flank of mouse subcutaneously for tumor growth. The in vivo-in vitro system LA795 Vv-Vt is an excellent model dissecting and analyzing the various factors which affect tumor development and determine the response of tumor to specific agent and regimens.

[In vivo experiments, in vitro assays in experimental oncology: establishment of LA 795 Vv-Vt].

In vivo experiment, in vitro assays (in vivo-in vitro system) is a kind of experimental technique which is different from both in vitro cell line and tumor line. The new model can be grown and studied either as an animal tumor or a cell culture. The specimen could be assayed for cell survival in vitro. This model can be used to study the response of malignant cells to the treatment in a precision and depth that is impossible in tumors grown in vivo. LA 795 Vv-Vt is the first in vivo-in vitro system developed in China. It was established with lung adenocarcinoma of T-739 mouse. The tumor cells could grow freely both in vivo and in vitro with a plating efficiency of 20%. Characteristics of LA 795 Vv-Vt tumor and cell in culture were described and compared. In order to estimate the radiation response of different doses, cell survival curves of tumors irradiated under oxic and hypoxic conditions were drawn and compared. The oxygen enhance ratio was 2.98. The experiments indicate that in vivo-in vitro system has a good dependent relation in dose and effect and is worth extensive application.