OBJECTIVETo investigate whether the major histocompatibility complex class I chain-related gene A gene (MICA) polymorphism and serum soluble MICA level were associated with the occurrence and development of colorectal cancer.

METHODSDNA samples from 117 colorectal cancer patients and 113 healthy individuals from Yangzhou in Jiangsu province were genotyped by using the polymerase chain reaction (PCR) and sequence-specific primer (SSP) method and PCR based sequencing. In addition, polymorphism at position 129 was also analyzed by PCR-SSP. Serum levels of soluble MICA were measured by a sandwich ELISA method.

RESULTSNeither the extracellular nor the transmembrane region polymorphisms of MICA gene were associated with the occurrence and the different stages of colorectal cancer. In contrast, the frequency of the methionine residue at position 129 was significantly decreased in the patient group. Soluble MICA levels in sera were increased in the late stages of colorectal cancer.

CONCLUSIONAlthough there was no genetic susceptibility attributed to MICA gene polymorphism with regard to development of colorectal cancer, serum levels of soluble MICA may be a diagnostic marker of advanced stages.

Colorectal Neoplasms ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Histocompatibility Antigens Class I ; blood ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

The activity of nano carbon fullerene lipidosome (NCFL) against influenza virus HINI in vitro was studied by observing the cytotoxicities and its activity rendered by different intensities of lighting with various periods of time. Rimantadine hydrochloride was used as the positive control drug. By using microcultural technique, the morphological changes of cells were observed and by using the gentian violet staining, antiviral activity of the NCFL against influenza virus was assayed. The results showed that: (1) The maximal concentration of the NCFL was 7μg/mL and the 50% toxic concentration (TC50) was 13.54μg/mL respectively; (2) NCFL had a significant activity of directly killing the influenza virus, while the activities in antiadsorption and antireplication were not obvious; (3) There was a dose-activity relationship between the dosages of NCFL and the direct killing effect against the influenza virus, and the periods of lighting-time could influence the activity partly. It was concluded that NCFL had a significant activity of directly killing the influenza virus.

To establish SP2/0 cell line H-2(d) stably expressing bcr-abl fusion gene fragment, the bcr-abl fusion gene was subcloned into retroviral vector pLXSN from pGEMbcr-abl. The recombinant retroviral vector pLXSNbcr-abl was transfected into PT67 packaging cells with the help of lipofectamine. The positive clones were selected out and cultured after G418 selection. Then viral supernatant was collected to determine viral titer, the viral titer was 2 x 10(7) CFU/ml. The SP2/0 cells were infected with the collected viral supernatant. The results showed that after G418 selection, the bcr-abl fusion gene was integrated into the chromosome of SP2/0 cells infected stably, with recombinant retrovirus and expressed in SP2/0 cells confirmed by PCR and RT-PCR respectively. In conclusion, the mouse tumor cell lines expressing the bcr-abl fusion protein were successfully established and would be used as a experimental cell model for anti-CML immunotherapy.
Animals ; Cell Line ; Cell Line, Tumor ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Mice ; Mice, Inbred BALB C ; Multiple Myeloma ; genetics ; pathology ; NIH 3T3 Cells ; Peptide Fragments ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods

OBJECTIVETo study the specific immune response induced by a recombinant eukaryotic expression plasmid encoding bcr-abl fusion gene fragment so as to explore new immunotherapy in mouse.

METHODSA recombinant eukaryotic vector pVbcr-abl expression cDNA fragment of bcr-abl fusion gene was constructed and used to immunize BALB/c mice. Serum level of bcr-abl specific antibody was detected by enzyme-linked immunosorbent assay (ELISA). Twenty days later the immunized mice were subcutaneously inoculated SP2/0/bcr-abl cells. The survival time, tumor growth time and lymphocytic infiltration were observed. T cells infiltration into tumor tissue was analyzed by immunohistochemistry. Changes of T cell subset in the spleen of mice was analyzed by fluorescent-activated cell sorting (FACS) and the cytotoxicity T lymphocyte (CTL) activity in spleen by lactate dehydrogenase (LDH)-release assay.

RESULTSThe eukaryotic expression vector pVbcr-abl was constructed successfully, and highly expressed the cDNA fragment of bcr-abl fusion gene. The BALB/c mice immunized with the vector could generate the specific antibody and CTL, resulting in a specific immunoprotection. There were dramatic differences in the tumor-forming time, tumor ulcer appearing time and tumor-growing speed between the immunized and the control groups. The mice had longer survival time in the immunized group than in the control group. There were a large amount of CD3(+) T cells infiltration in tumor tissue of the immunized mice. The spleen cells from the immunized mice had higher CTL activity with a alteration of T cell subset, the CD4(+)/CD8(+) ratio being 1.54 +/- 0.29, higher than that of control group (1.18 +/- 0.30).

CONCLUSIONThe recombinant eukaryotic expression plasmid pVbcr-abl can induce in vivo not only the generation of specific antibody, but also high level of specific CTL activity, resulting in killing the SP2/0/bcr-abl tumor cells directly and inhibiting the tumor growth.

Animals ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Genetic Vectors ; Immunotherapy ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Random Allocation ; Transfection

To study the influence of vaccine of bcr-abl fusion gene fragment on inoculated SP2/0/bcr-abl tumor cells in mice, BALB/c mice were immunized with pVbcr-abl, pVbcr-abl/mIL7 plasmids, respectively, then SP2/0/bcr-abl cells expressing the fragment of bcr-abl fusion gene were inoculated subcutaneously into the groin of BALB/c mice in order to observe the effect of vaccine on growth of inoculated SP2/0/bcr-abl tumor cells. The results showed that there were distinct differences on the time of tumor growth, the time of tumor ulceration, tumor volume and survival time of mice bearing tumor between two immunized groups and two control groups (blank and vacant plasmid groups). The mice immunized with pVbcr-abl/mIL7 lived longer as compared to mice immunized with pVbcr-abl. The tissue of inoculated tumor was more compact, tumor organ was larger, tumor form was irregular in 2 control groups, while the tissue of inoculated tumor was looser, tumor volume was smaller, and with mass inflammatory infiltration in two immunized groups. Moreover, the metastatic tumor cells were found in the livers of control groups, but not observed in two immunized groups. It is concluded that the protection occurred in immunized mice which inhibited the growth of SP2/0/bcr-abl tumor cell in vivo.
Animals ; Cancer Vaccines ; immunology ; metabolism ; Cell Line, Tumor ; Female ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Multiple Myeloma ; genetics ; immunology ; pathology ; Neoplasm Transplantation ; Random Allocation ; Vaccines, DNA ; immunology

OBJECTIVETo investigate the influence of mIL-7 on the immune response induced by vaccine of bcr-abl fusion gene fragment in mouse.

METHODSBALB/c mice were immunized by i. m. injection of pVbcr-abl/mIL-7 and pVbcr-abl, respectively. The specific antibody to p210bcr-abl protein was assayed by ELISA. The CTL activity of spleen cells from the immunized mice was assessed with LDH release test.

RESULTSThe pVbcr-abl/mIL-7 and pVbcr-abl-immunized BALB/c mice elicited higher specific antibodies to p210bcr-abl protein. The specific antibody level of former group was higher than that in latter group, but the difference was statistically not significant. The spleen cells from the immunized mice showed more effective CTL activity than that from control group. The cytotoxic activity of spleen CTLs induced by pVbcr-abl/mIL-7 immunized mice exceeded that of pVbcr-ab-immunized mice.

CONCLUSIONThe mIL-7 may influence the growth and differentiation of T cells, promote some T cells migrating into tumor tissue and up-regulate the specific cellular immune response. The results of this study provided an useful experimental basis for preclinical research on gene vaccine for chronic myeloid leukemia.

Animals ; Antibodies ; blood ; Cancer Vaccines ; genetics ; immunology ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; immunology ; Humans ; Interleukin-7 ; biosynthesis ; genetics ; immunology ; K562 Cells ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Spleen ; cytology ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccination ; Vaccines, DNA ; immunology

OBJECTIVETo compare the incidence of metabolic disorders (MS) in patients with primary aldosteronism (PA) and essential hypertension (EH).

METHODSMS prevalence was observed in 200 EH patients (male 104) and 220 PA patients (male 117) hospitalized to our hospital from August 2005 to March 2007.

RESULTS(1) The prevalence of MS in PA group was significantly higher than that of EH group (47.3% vs. 31.5%, P = 0.009). (2) Blood pressure was significantly higher in PA group than that of EH [SBP: (150.67 +/- 15.45) mm Hg vs. (145.69 +/- 17.13) mm Hg, P = 0.042; DBP: (93.03 +/- 10.51) mm Hg vs. (85.83 +/- 14.44) mm Hg, P = 0.037]. (3) Incidences of abdominal obesity (86.8% vs. 78.5%, P = 0.024) and insulin resistance (insulin sensitivity index: 42.42 +/- 16.11 vs. 49.58 +/- 22.43, P = 0.008) were significantly higher in PA group than in EH group.

CONCLUSIONThe prevalence of MS in hospitalized PA patients was significantly higher than that of EH patients characterized by prevalent abdominal obesity, insulin resistant and severe hypertension.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Hyperaldosteronism ; epidemiology ; metabolism ; Hypertension ; epidemiology ; metabolism ; Incidence ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged

OBJECTIVETo study the genetic polymorphism of 15 short tandem repeat (STR) (D2S1338ì D3S1358ì D5S818ì D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, CSF1PO, TPOX, TH01, vWA, FGA) in Maonan minority of Guangxi province.

METHODSThe allele frequencies and the genotype of 15 STR loci were analyzed in 143 unrelated individuals in Maonan minority of Guangxi by PCR-STR and genescan.

RESULTSThere were 130 STR alleles and 390 genotypes in the 15 STR of Maonan minority, with allele frequencies ranging from 0.0035 to 0.5385. The average heterozygosity was 0.7697, the discrimination power was higher than 0.8 except for that of TPOX, the accumulative discrimination power was more than 0.999999999, and the probability of paternity exclusion was more than 0.99999918.

CONCLUSIONThe 15 STR loci of Maonan minority in Guangxi possesses the characteristics of high genetic diversity, except for the TPOX locus. They can be employed in minority genetics investigation, individual and paternity test in forensic medicine.

Adult ; Aged ; Aged, 80 and over ; Alleles ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo evaluate the performance of 18F-FDG three-head tomography with coincidence imaging and serum tumor marker assays in identifying lung lesions in 104 patients with abnormal findings on chest X-ray or computer tomography.

METHODSA prospective evaluation of 18F-FDG coincidence imaging and the measurement of 3 serum markers for lung cancer ( carcinoembryonic antigen, CYFRA21-1 and neuron specific enolase) were performed within one week in 104 inpatients with suspected lung malignancy. All images were analyzed visually. It was considered positive for malignancy if the 18F-FDG uptake was increased relative to that in the adjacent lung tissue, and was focal. The serum tumor marker test was considered positive for malignancy if the serum level of at least one marker was elevated.

RESULTS66 patients were proven to have lung cancer by pathology, and 38 patients had benign lung diseases. The sensitivity, specificity, accuracy of 18F-FDG coincidence imaging and serum tumor markers in assessing lung cancers were 80. 0% , 77. 2% , 77. 9% and 56. 0% , 60. 9%, 64. 4% , respectively. 18F-FDG coincidence images in assessing lung lesions showed significantly higher sensitivity, specificity and accuracy than serum tumor markers. Four patients with lung cancer had negative findings on 18F-FDG coincidence images but showed positive serum markers.

CONCLUSION18F-FDG coincidence imaging is a powerful tool for evaluating patients with lung lesions suggestive of malignancy. Although the determination of serum marker levels is less accurate than 18F-FDG coincidence imaging, the combination of a positive 18F-FDG coincidence result and positive tumor markers may be helpful in improving the diagnosis of lung cancers.

Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm ; blood ; Biomarkers, Tumor ; blood ; Carcinoembryonic Antigen ; blood ; Carcinoma, Small Cell ; blood ; diagnosis ; diagnostic imaging ; Carcinoma, Squamous Cell ; blood ; diagnosis ; diagnostic imaging ; Female ; Fluorodeoxyglucose F18 ; Humans ; Keratin-19 ; Keratins ; blood ; Lung Neoplasms ; blood ; diagnosis ; diagnostic imaging ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; blood ; Plasma Cell Granuloma, Pulmonary ; blood ; diagnosis ; diagnostic imaging ; Positron-Emission Tomography ; Prospective Studies ; Radiopharmaceuticals ; Sensitivity and Specificity ; Tuberculosis, Pulmonary ; blood ; diagnosis ; diagnostic imaging