SSAO inhibitors suppress hepatocellular tumor growth in mice.

Vascular adhesion protein-1 (VAP-1) is both an endothelial adhesion molecule involved in leukocytes emigration, and an oxidase belonging to the family of semicarbazide-sensitive amine oxidases (SSAOs). The enzyme activity of VAP-1 plays an important role in the migration of myeloid-derived suppressor cells (MDSCs) into tumor site, and SSAO inhibitors can block the function of VAP-1. The effects of SSAO inhibitors on leukocyte infiltration and tumor progression were evaluated in H22 hepatocellular carcinoma-bearing C57BL/6 mice. Tumor weight and volume were measured after SSAO inhibitor treatment. Then, MDSCs recruitment and neo-angiogenesis were determined using immunostaining. SSAO inhibitors significantly blocked the catalytic activity of VAP-1 in tumor, attenuated tumor progression, and reduced neo-angiogenesis. CD11b(+) and Gr-1(+) MDSCs, which normally infiltrate into tumors, were significantly diminished in tumor-bearing mice treated with SSAO inhibitors. The present study demonstrated that SSAO inhibitors might have an anti-tumor effect on hepatocellular carcinoma by inhibiting recruitment of CD11b(+) and Gr-1(+) cells and hindering angiogenesis, which could be attributed to impairing the catalytic activity of VAP-1.

Lysine-specific demethylase 1 in breast cancer cells contributes to the production of endogenous formaldehyde in the metastatic bone cancer pain model of rats.

BACKGROUND: Bone cancer pain seriously affects the quality of life of cancer patients. Our previous study found that endogenous formaldehyde was produced by cancer cells metastasized into bone marrows and played an important role in bone cancer pain. However, the mechanism of production of this endogenous formaldehyde by metastatic cancer cells was unknown in bone cancer pain rats. Lysine-specific demethylase 1 (LSD1) is one of the major enzymes catalyzing the production of formaldehyde. The expression of LSD1 and the concentration of formaldehyde were up-regulated in many high-risk tumors. OBJECTIVE: This study aimed to investigate whether LSD1 in metastasized MRMT-1 breast cancer cells in bone marrows participated in the production of endogenous formaldehyde in bone cancer pain rats. METHODOLOGY/PRINCIPAL FINDINGS: Concentration of the endogenous formaldehyde was measured by high performance liquid chromatography (HPLC). Endogenous formaldehyde dramatically increased in cultured MRMT-1 breast cancer cells in vitro, in bone marrows and sera of bone cancer pain rats, in tumor tissues and sera of MRMT-1 subcutaneous vaccination model rats in vivo. Formaldehyde at a concentration as low as the above measured (3 mM) induced pain behaviors in normal rats. The expression of LSD1 which mainly located in nuclei of cancer cells significantly increased in bone marrows of bone cancer pain rats from 14 d to 21 d after inoculation. Furthermore, inhibition of LSD1 decreased the production of formaldehyde in MRMT-1 cells in vitro. Intraperitoneal injection of LSD1 inhibitor pargyline from 3 d to 14 d after inoculation of MRMT-1 cancer cells reduced bone cancer pain behaviors. CONCLUSION: Our data in the present study, combing our previous report, suggested that in the endogenous formaldehyde-induced pain in bone cancer pain rats, LSD1 in metastasized cancer cells contributed to the production of the endogenous formaldehyde.

[Determination of serum copper and zinc in different chemical forms by graphite furnace atomic absorption spectrometry with ethanol-EDTA precipitation method].

A simple and reliable method was developed for the determination of serum copper and zinc in different chemical forms by graphite furnace atomic absorption spectrometry (GFAAS) with ethanol-EDTA precipitation. The serum and ethanol were mixed with volume ratio 1 : 2. The mixture was incubated at 70 degrees C and ultra-centrifuged to precipitate proteins. Zinc and copper can be released from albumin after EDTA treating. Thus, macroglobulin-zinc, ceruloplasmin-copper, and albumin-bound zinc or copper could be determined by two proposed precipitation steps. The determination limit of copper (3sigma) was 1.2 microg x L(-1) and the recovery was 92.3%-104%, while the determination limit of zinc (3sigma) was 0.098 microg x L(-1), and the recovery was 90%-107%. This two-step precipitation method can be used to determine serum zinc and copper in various chemical forms in tumor, Wilson patients and heathy human.

[Determination of trace free copper and zinc in serum samples by graphite furnace atomic absorption spectrometry after two-step precipitation of protein].

A method was developed for the determination of trace free copper and zinc in serum sample by GFAAS after two-step precipitation. The serum and ethanol were mixed with volume ratio of 1 : 2. The mixture was subsequently denatured at 70 degrees C and centrifuged to precipitate proteins. The determination, limit of copper (3sigma) was 1.2 microg x L(-1) and the recovery was 92.3% - 104%, and the determination. Limit of zinc(3sigma) was 0.098 microg x L(-1) and the recovery was 90%-107%. This two-step precipitation method can be used to determine non-protein-bound copper in tumor patients and healthy people.

[Determination of trace lead in water samples and salt samples by graphite furnace atomic absorption spectrometry after cloud point extraction].

A method was developed for the determination of trace lead in water samples and salt samples by GFAAS after cloud point extraction. The parameters of extraction system such as pH, the concentrations of the extractant and the surfactant, and the time for cloud point extraction were optimized. Under the optimized conditions, the detection limits of lead were 0.000 5 microg x g(-1) for salt, and 0.01 microg x L(-1) for water, respectively. The proposed method was applied to the determination of lead in water samples and salt samples, and satisfactory results were obtained.

[Study of protein conformation in solution by tyrosine residues RRS spectrum].

Due to the interference caused by the emission of tryptophan residue, it is hard to use fluorospectrophotometry to detect the spectrometric changes of the tyrosine residue when protein conformation is changed. When the concentration of protein in solution is relatively high, tyrosine residue has a characteristic scattering peak when excited with its K-band wavelength of light. In the present study, the authors established a method for detecting protein conformation changes in solution through the changes (peak height and wavelength shift) of the characteristic scattering peak of tyrosine residue. The method may be used for detecting protein conformation changes in solution caused by the changes of electrolyte, pH, and temperature.

[Determination of trace cobalt in water samples by graphite furnace atomic absorption spectrometry after cloud point].

In buffer solution of citric acid monohydrate-disodium hydrogen phosphate at pH 5-7.5, cobalt water samples were chelated by 1-(2-pyridylazo)-2-naphthol (PAN) to form Co-PAN. After water-bath at 66 degrees C for two hours, Co-PAN is extracted into Triton X-100 nonionic surfactant phase and separated from bulk water. Extracted cobalt content was measured by graphite furnace atomic absorption spectrometry. The surfactant phase separated was treated with 0.5% HNO3-0.1% Pd(NO3)2 to remove background interference at 1200 degrees C temperature. The pre-concentration of cobalt in water samples permitted the detection of 0.003 microg x L(-1) (10sigma) with the enhancement factor of 100. The recoveries were 90.5%-106%. The proposed method was applied to the determination of cobalt in water samples and satisfactory results were obtained.

Intratumoral injection of arsenic to enhance antitumor efficacy in human esophageal carcinoma cell xenografts.

To enhance the therapeutic efficacy of anticancer agents and to reduce systemic side effects, it was decided to study the effect of arsenic trioxide directly on solid tumors to observe the anticancer effect of arsenic on tumors and the distribution of arsenic in tumors and other organs. Esophageal carcinoma cells were heterotransplanted in severe combined immunodeficient (SCID) mice in both laterals of the abdominal wall. When both lateral tumors had grown to approximately 10x8x5 mm3, tumor-bearing mice were used for 2 experiments. The right tumors were treated with intratumoral injection of As2O3 in 1, 5 and 10 micro g per day for 10 days sequent. The left tumors were treated with phosphate buffer solution as controls. To explore the distribution of As2O3 remaining in tumor and some organs, a single intratumoral injection of As2O3 was studied with quantitative measurement of arsenic by means of atomic absorption spectrometry. The results revealed that on the 17th day after the 1st injection As2O3-treated tumors were suppressed markedly compared to that of the contrarily lateral tumor accompanied by marked apoptosis and necrosis in tumor cells. The tumor growth inhibition (TGI) was 13.56, 62.37 and 76.92% in 1, 5 and 10 micro g group, respectively. There were no pathological changes in heart, lung, spleen, liver, kidney or brain after arsenic administration. Distribution of As2O3 revealed that As2O3 remained at higher concentration in arsenic-treated tumor tissue than in other organs. Our data suggest that intratumoral delivery of As2O3 efficiently suppresses growth of transplanted esophageal carcinoma without systemic side effects. The protocol of As2O3 intratumoral injection will be its potential clinical utility for therapy of solid tumors.

[Determination of arsenic in As2O3-treated esophagoscope-transplant mouse tissue by graphite furnace atomic absorption spectrometry].

As2O3 was injected into tumour tissue of esophagoscope-transplant mouse. The concentration of As diffusing into other tissues was investigated. A method was proposed for the determination of As in tissue samples of mouse by graphite furnace atomic absorption spectrometry. After wet digestion (water-bath at 80 degrees C) with 2:1:1 (psi) of HNO3-H2SO4-HClO4, the digested tissue samples of mouse were diluted with 0.2% Triton X-100-0.4% AgNO3. The matrix-matching calibration curve of non-interference was established with standard addition method. The relative standard deviation was 3.2%-8.7%. The limit of detection was 1.57 microg x L(-1). The recoveries were 81.7%-105% Arsenic concentrations in mouse liver, kidney, brain, left-chest, and right-chest were determined after injection of arsenic at different times.