Recent studies have revealed that sox4 gene expresses abnormally in many kinds of tumor tissues and it probably involves in tumorigenesis,development and metastasis of cancer.Regulating proliferation,differentiation,apoptosis and other process may participate in the work mechanisms of sox4 gene.Therefore,further studies about the relationship between sox4 gene and tumor would provide new ideas of exploring special diagnosis markers and novel targets for tumor therapy.

Objective To study the clinical and prognostic value of CK19 mRNA-positive circulating tumor cells in early breast cancer patients. Methods We analyzed the peripheral blood in 50 patients with early breast cancer after surgery and before the initiation of any adjuvant treatment for the presence of CK19 mRNA-positive circulating tumor cells using a nest reverse polymerase chain reaction assay. All patients were followed up. Results CK19 mRNA-positive cells were detected in 40.0 %(20/50) of patients with early breast cancer, 12.5 %(3/24) of patients with breast benign lesions, but 5 %(1/20) in healthy individuals (P =0.017,P =0.004); 11 to 20 of them relapsed during the follow-up period (P =0.002). There was no significant association between the detection of CK19 mRNA-positive cell and the patients' menstrual status, tumor stage, tumor size, etc (P ＞0.05). Detection of peripheral-blood CK19 mRNA-positive cells was associated with reduced median relapse-free interval in early breast cancer patients (P =0.007). Conclusion CK19 mRNA is one of the molecular markers for the detection of circulating tumor cells in early breast cancer. Detection of peripheral blood CK19 mRNA-positive cells might be an important predictive value as a marker of relapse in early breast cancer patients.

Objective To assess the effect of extracorporeal shock waves(ESWs)as a treatment for ⅢB chronic prostatitis(CP).Methods Forty-six men with ⅢB CP were randomly divided into an experimental group (n=34)and a control group(n=12).The patients in experimental group received low energy ESW treatment,20000 impulses in 10 sessions over 2 weeks.The patients in control group received sham ESW treatment without shock waves energy under the same other conditions as in experimental group.Pain,urination and quality of life/impact were assessed with National Institutes of Health-chronic prostatitis symptom index(NIH-CPSI).Both groups were assessed at baseline,post-treatment and at a 4-week follow-up.Resuits The total NIH-CPSI scores,pain scores and quality of life/impact scores in experimental group decreased significantly post-treatment(P＜0.01),but urination scores did not(P＞0.05).Similar decreases of these scores were also found in control group post-treatment.The total NIH-CPSl scores and pain scores maintained at a lower level at the 4-week follow-up in experimental group,but the scores returned to the level as pre-treatment in control group.The effectiveness rates and prominent effectiveness rates in experimental group were significantly higher than those in control group post-treatment and at the 4-week follow-up(all P＜0.05). Conclusions ESWs was effective in the treatment for ⅢB CP.After ESWs treatment pain alleviated,symptoms reducea and quality of life improved.

Objective To investigate the way of inducing dendritic cells from precursor in human cord blood and its role in antitumor immunity.Methods Cord blood was collected under sterile condition and the cord blood mononuclear cells were separated by centrifugal in density gradient.CBMCs were cultured with GM-CSF+IL-4+TNF-?and cell phenotype was analyzed with CD1a、CD83 antibody by using indirect immunofluorescence assays.The effects of DCs pulsed with tumoral antigens on cytotoxic T lymphocytes(CTLs) inducement and growth inhibition of YTMLC cells were assayed.Results Our results indicated that DCs precursors in human cord blood can be induced to differentiate in the medium containing GM-CSF、IL-4 and TNF-?.The cells with typical morphological properties of DCs were observed at the 7th day.At that time,(20.8?1.62)%CD1a+ cells were obtained.After incubation with tumor cytolysis antigen,the DCs can activate the CTLs to become tumor specialized CTLs,which had shown significantly inhibition on growth of YTMLC tumor cell line.Conclusion The precursors in human cord blood can be induced to functional DCs which activate T lymphocyte to become tumor specialized CTLs.

ObjectiveTo identify mutations ofGATA4 andGATA6 genes in children with isolated congenital atrial septal defect (ASD).Methods From November 2012 to November 2013, 101 patients with ASD (99 unrelated patients and one twin) who were submitted to catheter-based intervention and 100 ethnicity-matched children without congenital heart disease, blood disorders and chromosomal abnormalities were enrolled. The blood was collected. The coding regions and lfanking regions of theGATA4 andGATA6 genes were ampliifed by polymerase chain reaction and sequenced using the dideoxvnucleotide chain termination technique, and then compared with the normal sequence in the Genbank.Results Two novel heterozygous missense GATA6mutations, c. G145A and c. G151A, were identiifed in 2 unrelated ASD patients, which were not present in the controls. These two mutations predicted the conversion of glycine into serine at amino acid residue 49 (G49S) and glutamate into lysine at amino acid residue 52 (K52E). A heterozygous missenseGATA6 mutation c.43 G>C, which caused a conversion from glycine to arginine, was found in 9 ASD patients and 7 controls. A single nucleotide polymorphism c.99G>T, which did not cause amino acid conversion inGATA4 gene, was found.ConclusionsGATA6 gene is an important transcription factor in heart development. The mutation ofGATA6 gene may cause the change of its transcriptional activity, and lead to ASD.

GATA6 transcription factor belongs to the GATA family and contains 2 conserved zinc ifnger DNA binding domains. GATA6 not only presents in embryonic tissues but also found in heart, lung and pancreas and is essential for the maintenance of their function.The present review focuses on the critical roles of GATA6 in heart development and atrial septal defect to provide theoretical basis for diagnosis and treatment of atrial septal defect.

BACKGROUND AND OBJECTIVEThe mouse lung cancer orthotopic model includes spontaneous lung cancer model and endotracheal transplanted model, and etc. The spontaneous lung cancer needs longer time and does not ensure the rate of the generation of the tumor; as for endotracheal transplanted model, the position and size of the tumor are instable. In this study, the 3LL cell line was orthotopically transplanted into the lung of the C57BL/6 mice, compare to the heterotopic model, to discuss their stability and transfer-characteristics. And this study was also to optimize the method of establishing lung cancer orthotopic animal model.

METHODSDifferent quantity of 3LL cells were inoculated into the left oxter of C57BL/6 mice to establish the heterotopic model; or suspended with Matrigel then inoculated into the left lung of C57BL/6 mice to establish orthotopic model. The survival-time of the mice was examined. The tissue was collected for the subsequent histology assay after euthanizing the mice. Microvessels density (MVD) was observed and counted by immunohistological chemistry. CD44v was detected by flow cytometry.

RESULTSTTumor-form-rate of the heterotopic group were 100%, 66.7%, 16.7%, respectively, and had no macroscopic transfer. Tumor-form-rate of the orthotopic group were 100%, 100%, 83.3%, respectively, and had widespread transfer in contralateral chest and the lung. The median survival time of the orthotopic group (38, 35, 23 days) were less than the heterotopic group (82, 72, 50 days). MVD of the orthotopic group (120.2 +/- 9.73) was higher than the heterotopic group (92.6 +/- 7.12). The expression of CD44v of orthotopic (26.46 +/- 1.56)% was higher than the heterotopic group (23.13 +/- 1.02)%.

CONCLUSIONThe lung cancer orthotopic model which established by 3LL cells transplanted into the lung of the mice is simple, dependable, repeatable and has stronger transfer characteristics than the heterotopic model.

Animals ; Carcinoma, Lewis Lung ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Lung Neoplasms ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Random Allocation

Rhesus macaque (Macaca mulatta) is the best model to study of human immunodeficiency virus (HIV) infection and to develop acquired immunodeficiency syndrome (AIDS) vaccine. The crystal structure of its major histocompatibility antigen complex (MHC) is helpful to understand the mechanism of HIV immune evasion. In this study, we cloned the light chain (beta2m) of MHC class I allele of rhesus macaques, Mamu-A*02, and inserted it into pET21a(+) vector. We transfected the recombinant plasmid pET21a(+)-Mamu-beta2m and pET21a(+)-Mamu-alpha into BL21(DE3). Mamu-A*02 and beta2m were expressed in the form of inclusion bodies in BL21 (DE3). We co-refolded the inclusion bodies of Mamu-alpha and Mamu-beta2m with SIV nonapeptide YY9 and obtained the correct refolded protein complex. Then we purified the protein complex by the gel filtration and anion-exchange column. With hanging-drop method, we screened and optimized for the protein crystal. We managed to collect a X-ray diffraction with the resolution to 2.8 angstroms in the condition of 0.1 mol/L BIS-TRIS (pH5.5), 2.0 mol/L(NH4)2SO4. This crystal belong to perpendicular space group P2(1)2(1)2(1), with unit-cell parameters a = 128.99 angstroms, b = 129.01 angstroms, c = 129.03 angstroms. This data is available for the structure determination.
Animals ; Crystallography, X-Ray ; Epitopes ; immunology ; Escherichia coli ; genetics ; metabolism ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Macaca mulatta ; Oligopeptides ; biosynthesis ; genetics ; Simian Immunodeficiency Virus ; immunology

OBJECTIVETo evaluate the efficacy and safety of celecoxib in treating inflammatory(Type IIIA) chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS-IIIA type).

METHODSSixty-four patients with diagnosed CP/CPPS-IIIA were randomized equally into two groups, Group A treated with celecoxib 200 mg daily(qd), while Group B with 200 mg twice a day(bid), both for 6 weeks. The white blood cell (WBC) count in expressed prostate secretion(EPS) and National Institutes of Health Chronic Prostatitis Symptom Index(NIH-CPSI) were assessed and compared at baseline(0 week) and at 2, 4, 6 weeks or the endpoint.

RESULTSThe mean number of WBC in EPS and the mean NIH-CPSI total scores were decreased gradually after treatment from baseline in both groups. The mean number of WBC of in EPS of either group at the endpoint was decreased by 46.2% and 69.4% respectively(Group A vs Group B) compared with the baseline level. The mean NIH-CPSI total scores of the two groups were decreased respectively by 5.6 and 8.3 points (Group A vs Group B). In terms of the above two parameters, Group B, responded better than Group A to the treatment. The differences observed above were statistically significant(all P < 0.05). No serious adverse event presented.

CONCLUSIONCelecoxib is effective and safe for patients with CP/CPPS(IIIA). The dosage of 200 mg twice a day is more efficacious than that of 200 mg daily.

Adult ; Celecoxib ; Chronic Disease ; Cyclooxygenase Inhibitors ; therapeutic use ; Humans ; Male ; Middle Aged ; Prostatitis ; drug therapy ; Pyrazoles ; Sulfonamides ; therapeutic use

OBJECTIVETo study the effect of targeted Sox4 gene-knock-down on the growth of xenografts of Xuanwei female lung cancer cell line XWLC-05 cells in nude mice.

METHODSRecombinant plasmid pGFP-V-RS-Sox4 shRNA was constructed and transfected into XWLC-05 cells. Real-time quantitative PCR and Western blot were applied to confirm the effect of Sox4 gene-knock-down. XWLC-05 cells stably transfected with the plasmids were inoculated into nude mice to establish the xenograft model. The nude mouse status, tumor formation and tumor growth were observed, and the tumor inhibition rate was calculated. CT scan was performed to assess the metastasis of xenografts. Immunohistochemical staining was applied to detect Sox4 and ki-67 protein expression.

RESULTSRecombinant plasmid pGFP-V-RS-A-Sox4 shRNA which can effectively knocking-down Sox4 gene was successfully constructed and the stable transfected cells were selected by puromycin-screening. The success rate of tumor cell inoculation was 100% in the mice of all groups except those inoculated with saline. The body weight of all mice inoculated with parental XWLC-05 cells (blank control), pGFP-V-RS-scram shRNA trsfected XWLC-05 cells (negative control), and pGFP-V-RS-Sox4 shRNA transfected XWLC-05 cells was increased to a varying degree, but there was no significant difference among the groups (P > 0.05 ). The growth of xenografts was significantly inhibited after silencing the Sox4 gene expression when compared with that of the blank and negative controls (P < 0.05) . The volume of removed tumors of the Sox4 gene-inhibited mice was (2.30 ± 0.34) cm(3) , significantly smaller than that of the negative control (3.99 ± 0.45) cm(3) and the blank control (4.03 ± 0.42) cm(3) (P < 0.05) . The weight of removed tumors of Sox4 gene-inhibited mice was (0.86 ± 0.14) g, significantly lower than that of the negative control (1.84 ± 0.27) g and blank control (1.86 ± 0.22) g, (P < 0.05). Immunohistochemical staining showed that Sox4 and ki-67 proteins mainly expressed in cell nuclei. The staining was significantly decreased in xenografts of Sox4-inhibited mice when compared with the negative and blank controls (P < 0.05). No distant metastasis was found in any mouse by CT imaging and pathological examination during the observation period.

CONCLUSIONSThe xenograft model of Xuanwei female lung cancer cell line XWLC-05 cells in nude mice is successfully established. Knocking-down of Sox4 gene can suppress the xenograft tumor growth.

Animals ; Female ; Gene Knockdown Techniques ; Heterografts ; Humans ; Lung Neoplasms ; genetics ; pathology ; Mice ; Mice, Nude ; SOXC Transcription Factors ; metabolism