Animals ; Biomarkers, Tumor ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Neoplasm Proteins ; secretion ; Neoplasms ; etiology ; pathology ; secretion ; Proteome ; secretion ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Adenosine Triphosphate ; metabolism ; Colorectal Neoplasms ; diagnosis ; genetics ; DNA Methylation ; Diphosphates ; metabolism ; Glioma ; diagnosis ; genetics ; Humans ; Isocitrate Dehydrogenase ; genetics ; Janus Kinase 2 ; genetics ; Mutation ; Myeloproliferative Disorders ; diagnosis ; genetics ; Polymorphism, Single Nucleotide ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins p21(ras) ; Sequence Analysis, DNA ; methods ; Templates, Genetic ; ras Proteins ; genetics

U251 cell is a sensitive cell line to serum, which stops at G0 phase of cell cycle in no-serum medium, and recovers growth when the serum is added into no-serum medium. The cell can express corresponding proteins in different phase of cell cycle. Therefore it is very signification for the study of cell cycle regulation mechanism that explores these proteins. In this paper, the mouse antibody phage display library was added into the bottle in which the serum starvation U251 cells had been cultured, and the special antibody phages were absorbed. Then the absorbed antibody phages were amplified by adding E. coli TG1 and helper phage M13K07. Amplified antibody phages were added into bottle in which the serum cultured cell after serum starvation (follow named as serum recovered cells) were incubated, so that the cell absorbed the no-special antibody phages for the serum starvation cell and the special antibody phages were in supernatant. The remaining no-special antibody phages in the supernatant were discarded by repeating above program 3-4 times. The pure special antibody phages were gotten, and amplified by adding the host cell E. coli TG1 and helper phage M13K07. Then the host bacterium infected special antibody phage was spread on the plate medium with ampicillin, and the monoclonal antibody phages were gotten. Using same as above program, the monoclonal antibody phages absorbed specially for serum recovered U251 cells were obtained when the serum recovered cells instead of serum starvation cells and serum starvation cells instead of serum recovered cells. In this study, ninety-six positive monoclonal antibody phages that absorbed specially the serum starvation cells and eighty-two positive monoclonal antibody phages that absorbed specially the serum recovered cells were obtained. By using cell immunochemistry assay, two special signification antibodies were obtained. one (No.11) was the strong response in serum starvation cells, the other (No.2) was the strong response in serum recovered cells. The antibody No.2 had the distinctive response to the serum recovered cells in different incubation time (15min, 30min, 1h, 2h, 4h, 8h, 12h and 48h) after serum starvation. The results showed that No.2 antibody would be useful to research the factors of cell cycle regulation and apply to tumor diagnosis.
Antibodies, Neoplasm ; genetics ; immunology ; isolation & purification ; Bacteriophages ; genetics ; Brain Neoplasms ; immunology ; pathology ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Glioma ; immunology ; pathology ; Humans ; Peptide Library

Objective To investigate the effect of seafood intake on the urinary iodine level in women for exploring an alternative to iodine supplementation.Methods Healthy pregnant women and non-pregnant women, aged 20～40 years,were selected during their health examination in local women'S health care in 2006.The types of seafood and its intake frequency were recorded from these women.and urine and kitchen salt samples were collected for iodine determination.Results A total of 198 women including 148 pregnant and 50 non-pregnant women were recmitod for this study;they had a median level of urine iodine of 87.51 mg/L.The median levels of urine iodine of83.49,91.52,166.45μg/L in three group women classified as hardly,seldom and often intake of see food showed significant difference(X2=6.202,P<0.05).Urine iodine level in non-pregnant women taking seafood (90.94μg/L)was higher than that in pregnant women(84.79μg/L),the difference being statistically significant (U=3318.00,P<0.05).The urine iodine in pregnant women with seldom intake of seafood(94.46 μg/L)was significantly higher than that in the hardly intake women(83.28 μg/L),the difference being statistically significant (U=1257.5,P<0.05).During late period of gestation,the urinary iodine in the women ofthree statUS of hardly. Seldom and often intake of seafood were 81.93,97.97 and 140.18 μg/L,respective,with significant differences among them.Conclusions A certain amount of seafood taken every week Can increase urine iodine levels,and a direct relationship Was observed.Therefore,we suggest that it is necessary to advocate taking seafood to pregnant women for prevention of cretinism,particularly in the air.as where iodized salt was difficult to implement.

OBJECTIVETo study the clinicopathologic features of sclerosing angiomatoid nodular transformation of spleen and its differential diagnosis.

METHODSThe clinicopathologic characteristics and immunophenotype of 4 cases of sclerosing angiomatoid nodular transformation of spleen were studied.

RESULTSHistologically, all cases were characterized by multiple angiomatoid nodules of various sizes in a fibrosclerotic stroma. The nodules were round and sometimes convoluted. They were composed of slit-like, irregular-shaped or slightly dilated vascular spaces lined by plump endothelial cells and interspersed with a population of spindly or ovoid cells. Immunohistochemical study showed a heterogeneous staining pattern, with the lining cells of the small capillaries expressing CD34 and those of the sinusoid-like structures expressing CD8. CD31 highlighted both the lining cells and interspersed cells, resulting in a complex meshwork. The lining cells were also focally positive for CD68. Smooth muscle actin revealed conglomerates of spindly shaped cells around and between the vascular channels. These spindly shaped cells in the intervening stroma were focally positive for actin, but negative for desmin, CD21 and CD35.

CONCLUSIONSSclerosing angiomatoid nodular transformation is a rarely encountered benign lesion of the spleen, which should be distinguished from other angiomatoid tumors and tumor-like lesions.

Adult ; Angiomatosis ; metabolism ; pathology ; surgery ; Antigens, CD34 ; metabolism ; CD8 Antigens ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Hamartoma ; pathology ; Hemangioma ; pathology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Sclerosis ; pathology ; Spleen ; pathology ; Splenectomy ; Splenic Diseases ; metabolism ; pathology ; surgery ; Splenic Neoplasms ; pathology

OBJECTIVESTo construct a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 (MMP2) and to study its inhibitory effects on the invasiveness and migratory capacity of hepatocellular carcinoma (HCC) cell line HepG2 in vitro.

METHODSTotal RNA was extracted from HCC. Then a 500 bp fragment at the 5' end of the human MMP2 cDNA sequence was synthesized by polymerase chain reaction (PCR) and was reversely inserted into the multiclone site (MCS) of the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.coli BJ5183 and the recombinant adenoviral plasmid carrying the antisense MMP2 gene was constructed. The adenovirus (Ad-MMP2AS) was packaged and amplified in the HEK 293 cells and the viral titer was checked by GFP. Using the Boyden chamber model, the influence of Ad-MMP2AS on the invasion ability of HepG2 cells was determined in vitro.

RESULTSThe recombinant adenovirus vector carrying antisense MMP2 was constructed successfully and a strong green fluorescence was observed in HepG2 cells under a fluorescence microscope. The viral titer was 1 x 10(8); Ad-MMP2AS can effectively inhibit the penetrating capacity of HepG2 cells through Matrigel in vitro.

CONCLUSIONThe recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and migratory capacity of HepG2 in vitro and may have potential in treating HCC.

Adenoviridae ; genetics ; Carcinoma, Hepatocellular ; pathology ; Genetic Vectors ; Humans ; Liver Neoplasms ; pathology ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; pharmacology ; Neoplasm Invasiveness ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo study the extent and incidence of sensorineural hearing loss (SNHL) after radiotherapy (RT).

METHODSTwenty-eight patients with diagnosed nasopharyngeal carcinoma (NPC) were selected. The pure-tone audiography, auditory brain stem evoked response (ABR), impedance audiometry and evoked otoacoustic emissions (EOAE) recordings were performed before RT, 1 month, 1, 2 and 5 years after RT.

RESULTSAt 1 month after RT, there were 7.1 and 25.7 dB increased mean bone conduction (BC) thresholds at speech (0.5 - 4.0 kHz) and at high frequency (8.0 kHz), and their BC thresholds were statistically significant increase than those before RT, respectively (P < 0.001). At 1 year after RT, there were 17.6 and 28.1 dB increased respectively, and their thresholds were statistically significant increase than those at pre-irradiation (P < 0.001). There were also significant increases in thresholds than those at 1 month of post-irradiation (P <0.001 or P < 0.05). At 2 years after RT, 21 and 27.4 dB were increased at respective those two frequencies, and there was a statistically difference only at speech frequencies when compared with those at 1 year after RT (P < 0.05). At 5 years after RT, 26.7 and 35.8 dB were increased at these two frequencies, and there were significant increases in threshold than those before, 1 month, 1 and 2 years after RT, respectively (P < 0.001). From 1 month to 5 years after RT, 37. 5% to 94. 7% of ears had a BC hearing threshold of at least 15 dB losses at speech frequency, whereas the percentage at high frequency was 85.4 to 97.4%. Up to 63.2% and 73.7% of ears had 30 dB SNHL at least at speech and high frequency, respectively. Furthermore, the degree of mean threshold loss was greater at high frequency than at speech frequency. The mean value of wave I, III and V latency, and I -V interpeak latency intervals of ABR had no significant difference between at 1 month after RT and before RT (P > 0.05). The wave I , III and V latency, and I - V interpeak latency intervals at 1 year and 2 years were significantly prolonged when compared with those before and 1 month after RT (P < 0.05), but there were no significant difference between 1 year and 2 years after RT (P > 0.05). The wave I, III and V latency, and I -V interpeak latency intervals at 5 years after RT were also significantly longer than those before RT (P < 0.001). There were significant difference in wave I , III and V latency (P < 0.05), and no significant difference in wave I - V interpeak latency intervals (P > 0. 05) between 5 years after RT and 1 year or 2 years after RT. Seven of 10 ears at 1 year after RT and 4 of 7 ears at 5 years after RT had normal EOAE, but they all had abnormal ABR response.

CONCLUSIONSSNHL in NPC patients start soon after completion of RT, especially more commonly in high frequency. The incidence and the extent of hearing loss are increased with time of follow-up. The hearing impairment could occur in the cochlea and/or the retrocochlear auditory pathway, which show that the sensitivity of radiation damage may be different in different patient and anatomic site of auditory system.

Adult ; Audiometry ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Loss, Sensorineural ; etiology ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; radiotherapy ; Otoacoustic Emissions, Spontaneous ; Radiotherapy ; adverse effects

The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Protein-Tyrosine Kinases ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Recombination, Genetic ; Retroviridae ; genetics

OBJECTIVETo study the anti-tumor effects of Newcastle disease virus (NDV) strain D817 on human colon carcinoma model in nude mice.

METHODSThe nude mouse model of human colon carcinoma was established by subcutaneous inoculation of human colon cancer LOVO cells. The tumor-bearing mice were given PBS, 5-Fu, high-dose NDV D817, moderate-dose NDV D817 or low-dose NDV D817 via caudal vein injection. The tumor size and weight of mice were measured. The liver damages were examined by histopathology. Apoptosis and necrosis of tumor cells were detected by flow cytometry. The endotumoral content of TNF-alpha was detected using a mouse TNF-alpha ELISA kit. The live virus was detected by hemagglutination (HA) test.

RESULTSThe moderate-dose NDV D817 inhibited the tumor growth more apparently than 5-Fu. The tumor growth inhibition rate reached to 48.1%. The liver damage and the weight change caused by NDV were less severe. NDV D817 made an increased apoptosis index and induced production of TNF-alpha. Live virus was not detected in important organs except in the tumor of nude mice by HA test.

CONCLUSIONIn the anti-tumor process in nude mice bearing xenografts of human colon carcinoma, a suitable dose of NDV D817 is more safe and effective.

Animals ; Apoptosis ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; therapy ; Humans ; Liver ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Newcastle disease virus ; physiology ; Random Allocation ; Tumor Burden ; Tumor Necrosis Factor-alpha ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo study the effects of strontium ranelate on osteoporosis induced by simulated weightlessness in male rats.

METHODSTwenty-seven male SD rats were randomly divided into 3 groups, namely the normal control group (group A) and two groups of weightlessness simulated by tail suspension (groups B and C). The rats in group C were treated with strontium ranelate, and those in the other two groups were given the same dose of normal saline for 28 consecutive days.

RESULTSThe rats in group B showed significantly lower levels of alkaline phosphatase (ALP), bone mineral density (BMD) and bone mineral content (BMC) than those in group A (P<0.05), but serum calcium and phosphonium concentrations underwent no significant changes in the 3 groups (P>0.05). In the rats of group B, the trabeculae of the femur became thinner, fragile, and discontinuous with reduced quantity as compared with those in group A. The rats in group C had greater number of the trabeculae than those in group B with decreased resorption surface and more regular arrangement of the collagen fibers.

CONCLUSIONStrontium ranelate may produce beneficial effect on the bone microstructure, reduce bone loss and stimulate bone formation in male rats subjected to simulated weightlessness.

Animals ; Male ; Organometallic Compounds ; therapeutic use ; Osteoporosis ; etiology ; prevention & control ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thiophenes ; therapeutic use ; Weightlessness Simulation ; adverse effects