Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

AIMTo study the chemical constituents in the mycelia of Hypomyces sp..

METHODSSilica gel column chromatography was employed for the isolation and purification. Chemical and spectral methods were used to determine the structures of the isolated compounds.

RESULTSTwo compounds were isolated and identified as: hypomycin C (I) and hypomycin D (II).

CONCLUSIONCompounds I and II are new compounds.

Chromatography, Gel ; methods ; Fermentation ; Hypocreales ; chemistry ; Molecular Conformation ; Molecular Structure ; Mycelium ; chemistry ; Perylene ; analogs & derivatives ; chemistry ; isolation & purification ; Quinones ; chemistry ; isolation & purification

OBJECTIVETo investigate the expressions of TopI gene in small cell lung cancer cell line H446, and explore the influence of TopI on the chemosensitivity of the cell line to topotecan (TPT).

METHODSWestern blot was performed to detect the TopI expression in H446 cells. Lipofectamine 2000 was used for the transient transfection of H446 cells by siRNA, and the transfection efficacy was detected. TopI mRNA was analyzed by quantitative RT-PCR and TopI protein was detected by Western blot to selected effective siRNA. The drug-sensitivity to topotecan (TPT) was evaluated by MTT assay.

RESULTSTopI gene was expressed in H446 cells. Lipofectamine 2000 mediated the siRNA effectively (88.67%). Compared with its parental cells, RT-PCR results showed that TopI mRNAs in transfected cells were reduced by (95.7 +/- 1.6)%, (90.8 +/- 1.6)%, (96.1 +/- 2.7)% and (96.3 +/- 1.8)%, respectively, and decreased significantly at protein level. By MTT assay, the inhibition rate of TPT to H446 cells transfected by siRNA was lower than that of control group at same concentrations (P < 0.01).

CONCLUSIONsiRNAs can silence the expression of TopI and decrease the drug-sensitivity of H446 cells to TPT.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type I ; genetics ; metabolism ; Down-Regulation ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Topotecan ; pharmacology ; Transfection

The study was aimed to examine the B cell activating factor promoter polymorphism of the TNF family (BAFF)-871 C/T in patients with immune thrombocytopenic purpura (ITP) and to explore its correlation with ITP and the relationship between the blood platelet count of newly diagnosed patients with ITP and genotypes of BAFF-871 C/T polymorphisms. Alleles specific polymerase chain reaction (ASP-PCR) and agarose gel electrophoresis were used to identify polymorphisms -871 C/T of BAFF promotor in 133 ITP patients and 117 healthy controls, and determine the genotype of subjects. Meantime, the frequency of genotype and alleles were analyzed. The results indicated that out of 133 patients with ITP, 33.1% patients exhibited C/C, 42.1% patients were heterozygous C/T, and 24.8% patients were homozygous T/T. The corresponding frequencies in 117 healthy controls were 55.6% C/C, 33.3% C/T and 11.1% T/T. The allele frequency of T in ITP patients and healthy controls were 45.9% and 27.4% respectively. There was significant difference in the BAFF-871 C/T genotypic frequency between the ITP patients and healthy controls (p < 0.05). The allele frequency of T in ITP patients was higher than that in healthy controls. There was no significant difference in the blood platelet counts between the various genotype (p > 0.05). It is concluded that the polymorphism -871 C/T of BAFF promoter is correlated with the pathogenesis of ITP. However, there is no significant difference in blood platelet counts between the various genotype, so the polymorphism -871 C/T of BAFF promoter can not be referred as the analysis index for evaluating the severity of ITP.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alleles ; B-Cell Activating Factor ; genetics ; Case-Control Studies ; Child ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Purpura, Thrombocytopenic, Idiopathic ; genetics ; Young Adult

OBJECTIVETo analyze the polymorphisms of B cell activating factor (BAFF) gene and the plasma levels of BAFF in patients with idiopathic thrombocytopenic purpura (ITP), and to investigate their roles in the pathogenesis of ITP.

METHODSAlleles specific polymerase chain reaction (AS-PCR) and agarose gel electrophoresis were used to identify polymorphisms -871C/T of BAFF promotor in 133 ITP patients and 117 healthy controls. The plasma levels of BAFF were assayed by ELISA.

RESULTSIn ITP group, the frequency of C/C, C/T and T/T was 33.1%, 42.1% and 24.8%, respectively, the corresponding frequency in control group was 55.6%, 33.3% and 11.1%, respectively. The allele frequency of T in ITP and control groups was 45.9% and 27.4%, respectively. There was a significant difference in the BAFF -871C/T genotypic frequency between the ITP and control groups (P < 0.05). BAFF antigen in untreated ITP, treated patients and controls was 875.86 pg/ml, 502.59 pg/ml and 736.88 pg/ml, respectively, being also a significant difference among the three groups (P < 0.05). BAFF antigen in homozygous T/T was higher than that in homozygous C/C and heterozygous C/T, but the difference was not statistically significant (P > 0.05).

CONCLUSIONSOver expression of BAFF may be a risk factor for ITP patients. There is a correlation of the BAFF promotor polymorphism -871C/T with ITP, but the polymorphism does not affect the expression of BAFF.

B-Cell Activating Factor ; genetics ; Gene Frequency ; Humans ; Interleukin-4 ; Polymorphism, Genetic ; Purpura, Thrombocytopenic, Idiopathic ; immunology

Acrylamide is a common chemical material, extensively used in industry and scientific experiments. Recently, it has been reported that starchy food cooked at high temperature can produce acrylamide. Acrylamide monomer has several toxic effects and the extensive concern for its toxicity has arisen with the finding of acrylamide formation in some processed foods. Researches have shown that acrylamide monomer can cause reproductive toxicity, including toxic effects on male reproductive behavior, male reproductive endocrine function and spermatogenesis. The mechanisms may include the effects of acrylamide on Leydig cells, the formation of motor protein/ chromosomal/DNA alkylation and damage by oxidative stress.
Acrylamide ; toxicity ; Animals ; Genitalia, Male ; drug effects ; physiology ; Male ; Sexual Behavior, Animal ; drug effects ; physiology ; Spermatogenesis ; drug effects

BACKGROUNDGene chip array can differentiate isolated mycobacterial strains using various mycobacterium specific probes simultaneously. Gene chip array can evaluate drug resistance to isoniazid and rifampin of tuberculosis strains by detecting drug resistance related gene mutation. This technique has great potential for clinical application. We performed a retrospective study to investigate the capability of gene chip array in the rapid differentiation of species and detection of drug resistance in mycobacterium, and to evaluate its clinical efficacy.

METHODSWe selected 39 patients (54 clinical mycobacterium isolates), used gene chip array to identify the species of these isolates and detect drug resistance to isoniazid and rifampin in Mycobacterium tuberculosis isolates. Meanwhile, these patients' clinical data were analyzed retrospectively.

RESULTSAmong these 39 patients whose mycobacterium culture were positive, 32 patients' isolates were identified as Mycobacterium tuberculosis, all of them were clinical infection. Seven patients' isolates were identified as non-tuberculosis mycobacterium. Analyzed with their clinical data, only two patients were considered as clinical infection, both of them were diagnosed as hematogenous disseminated Mycobacterium introcellulare infection. The other five patients' isolates were of no clinical significance; their clinical samples were all respiratory specimens. Clinical manifestations of tuberculosis and non-tuberculous mycobacterial infections were similar. Isoniazid resistance was detected in two tuberculosis patients, while rifampin resistance was detected in one tuberculosis patient; there was another patient whose Mycobacterium tuberculosis isolate was resistant to both isoniazid and rifampin (belongs to multidrug resistance tuberculosis). The fact that this patient did not respond to routine anti-tuberculosis chemotherapy also confirmed this result.

CONCLUSIONSGene chip array may be a simple, rapid, and reliable method for the identification of most mycobacterial species and detection of drug resistance in Mycobacterium tuberculosis. It is useful in diagnosis, treatment, and hospital infection control of mycobacterial infections, and it may have a great potential for clinical application.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antitubercular Agents ; therapeutic use ; Female ; Humans ; Isoniazid ; therapeutic use ; Male ; Middle Aged ; Mycobacterium ; classification ; genetics ; pathogenicity ; Mycobacterium tuberculosis ; genetics ; pathogenicity ; Oligonucleotide Array Sequence Analysis ; methods ; Rifampin ; therapeutic use ; Tuberculosis, Multidrug-Resistant ; genetics ; Young Adult

OBJECTIVETo investigate the clinical value of real-time tissue elastography (RTE) in the diagnosis of breast cancer.

METHODSOne hundred and twenty patients with breast lumps (135 lesions) were examined with B-mode imaging, color Doppler flowing imaging (CDFI) and RTE. The elastogram was graded using 5-score evaluating method. The postoperative pathological diagnosis was used as gold standard, and the sensitivity, specificity and accuracy of RTE and two-dimensional ultrasonography combined with RTE in diagnosis of breast cancer were calculated.

RESULTWhen the score >4 was set for cut-off criteria of malignancy, the sensitivity, specificity and accuracy of RTE was 85.45%, 83.75% and 84.4%, respectively. While two-dimensional ultrasonography combined with RTE was used, the sensitivity, specificity and accuracy increased up to 100%, 95% and 97%, respectively.

CONCLUSIONRTE combined with two-dimensional ultrasonography can improve the validity in the diagnosis of malignant breast lesions.

Adolescent ; Adult ; Aged ; Breast Neoplasms ; diagnostic imaging ; pathology ; Computer Systems ; Diagnosis, Differential ; Elasticity ; Female ; Humans ; Image Enhancement ; instrumentation ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography, Mammary ; instrumentation

OBJECTIVESTo determine changes of insulin-like growth factor-1 (IGF-1) and IGF binding protein-3 (IGFBP-3) in serum samples from benign prostatic hyperplasia (BPH) patients and to evaluate the value of these molecules as possible etiologic factors for BPH.

METHODSThe serum IGF-1 and IGFBP-3 levels were measured with immunoradiometric assay(IRMA) in 64 cases of BPH and in 30 healthy subjects as controls. The cases of BPH were divided into 3 groups according to the prostate volume(PV). There were 18 cases(PV < or = 30 ml) in group A, 24 cases(PV31 approximately 50 ml) in group B, 22 cases(PV > or = 50 ml) in group C.

RESULTSThere were no statistical differences between the levels of Both IGF-1 and IGFBP-3 in BPH groups and healthy groups (both P > 0.05), but there were statistical differences among three groups of BPH. Both IGF-1 and IGFBP-3 levels in group C of BPH were significantly higher than those in group A (both P < 0.05). A positive correlation between the serum levels of IGF-1 and PV displayed(r = 0.58), as well as IGFBP-3 (r = 0.48).

CONCLUSIONSTogether these observations implicate IGF-1 and IGFBP-3 as important factors during the progression of BPH. It shows the value of non-operation treatment for this disease.

Aged ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; analysis ; Male ; Prostatic Hyperplasia ; blood ; Radioimmunoassay

Chromosomes, Human, Pair 22 ; Humans ; Infant, Newborn ; Male ; Trisomy ; pathology