Objective To evaluate the value of ultrasound elastography score and strain ratio index in the differential diagnosis of thyroid solid nodules. Methods Eighty-six thyroid solid nodules of 69 patients were divided into 2 groups based on their pathologic results: benign nodules group and malignant nodules group.Compared with pathological results, each thyroid solid nodules' elastographic scores and strain ratio index were respectively analyzed. The sensitivity, specificity and accuracy of elasticity score and strain ratio index were calctlated.The receiver operator characteristic (ROC) curves were drawn according to the results, to choose the best cut-off value.Results There were 67 nodules in benign nodules group and 19 nodules in malignant nodules group. The diameter of benign nodules was (2.05 ± 0.21)cm, malignant nodules was (1.62 ± 0.12)cm. There were no significant differences in the elasticity score and strain ratio index between benign and malignant nodules(t= - 1.736, P ＞ 0.05). Malignant lesions had a higher score and strain ratio index(median 3, 3.97, respectively) than benign ones(median 2, 1.66,respectively, Z = - 5.290, - 5.450, all P ＜ 0.05). If the cut-off value of the elastographic scores was equal or more than 3, the sensitivity, specificity and accuracy were 84.2%(16/19) ,86.6%(58/67) and 86.0%(74/86), respectively,with an area under ROC curve of 0.854. If the cut-off value of the strain ratio index was equal or more than 2.50,the sensitivity, specificity and accuracy were 89.4%(17/19),86.6%(58/67) and 87.2%(75/86), respectively, with an area under ROC curve of 0.911. Conclusion Elasticity score and strain ratio index are helpful in differentiating benign and malignant lesions of thyroid with high clinical value.

OBJECTIVERecombinant virus pulsated dendritic cells (DCs) may affect their survival, growth and maturity. This study is to test the infection efficiency of recombinant adeno-associated virus carrying hepatitis B core antigen (rAAV-HBV-c) to DCs and the growth and maturity of them.

METHODSPeripheral blood mononuclear cells (PBMCs) were isolated from healthy blood donors. Adherent monocytes were pulsed by rAAV-HBV-c and 293 lysate as controls on the first day of isolation. DCs were cultivated in AIM-V media with 1000 u/ml granulocyte macrophage stimulating factor (GM-CSF), 1000 u/ml interleukin-4 (IL-4) and 50 ng/ml tumor necrosis factor-alpha (TNFalpha) separately in vitro. DCs were examined at different times and the expressions of several clusters of differentiations (HLADR, CD14, CD80, CD83, CD86) were studied using FACS after being cultured for 7 days. The transcription and expression of HBV-C gene were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and intracellular staining fluorescence activated cell sorter (FACS), respectively.

RESULTSThe rAAV-HBV-c infected and uninfected monocytes gradually matured and their morphology had no significant differences. The CDs expressed on the surfaces of the two groups of DCs were also similar (HLADR: 96.1% vs. 94.5%; CD86: 87.7% vs. 89.8%; CD83: 75.6% vs. 78%; CD80: 52% vs. 54.3%; CD14: 6.4% vs. 4.5%). HBV-C gene mRNA expression was measured using RT-PCR and 89.5% of the rAAV-HBV-c infected DCs showed their protein expression using FACS.

CONCLUSIONrAAV-HBV-c can effectively pulse DCs without affecting the growth and maturity of them.

Cells, Cultured ; DNA, Recombinant ; genetics ; Dendritic Cells ; cytology ; immunology ; Dependovirus ; genetics ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Recombination, Genetic

OBJECTIVESAdeno-associated virus (AAV) Rep78 is known for its inhibitory effects on replication of several viruses and oncogenes transformations. The study was to investigate the effect of Rep78 on hepatitis B virus C (HBV-C) gene and the mechanism of it.

METHODSHBV-C promoter and HBV-C gene with its promoter were amplified by PCR and labeled with 32P-ATP. Electrophoretic mobility shift assay (EMSA) and in vitro transcription were utilized to detect the binding of MBP-Rep78 with HBV-C promoter and the transcription of HBV-C gene.

RESULTSEMSA showed that by increasing the amount of Rep78 protein from 0.1 microg to 1.0 microg, the binding bands got stronger in a dose-dependent manner. In addition, Rep78 antibody was used to certify the specificity of this binding. The compound of Rep78, Rep78 antibody and HBV-C promoter were seen as super shift bands in EMSA. Meanwhile, HBV-C gene transcription was significantly inhibited by in vitro transcription which meant that Rep78 could not only bind with HBV-C promoter, but also could inhibit the transcription of HBV-C gene.

CONCLUSIONAAV Rep78 could inhibit the transcription of HBV-C gene through its binding with HBV-C promoter.

DNA-Binding Proteins ; genetics ; Dependovirus ; genetics ; Gene Expression Regulation, Viral ; Hepatitis B virus ; genetics ; Humans ; Promoter Regions, Genetic ; genetics ; Transcription, Genetic ; Viral Proteins ; genetics

OBJECTIVETo elucidate the effects of sodium butyrate on rat hepatic oval cell differentiation in vitro.

METHODSHepatic oval cells were isolated from rats fed with a choline-deficient diet supplemented with 0.1% (w/w) ethonine for 4 to 6 weeks. The cultured hepatic oval cells were identified by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). After hepatic oval cells were treated with sodium butyrate, the morphological changes were studied through Giemsa staining and the albumin expression level was tested by Western blot.

RESULTSImmunohistochemical results showed the isolated cells were positive for both mature hepatocyte marker albumin and bile duct cell marker cytokeratin-19. Furthermore, RT-PCR results showed that the cells expressed stem cell marker c-kit, but not hematopoietic stem cell marker CD34. In short, the isolated cells were rat hepatic oval cells. 0.75 mmol/L sodium butyrate induced obvious phenotype changes of hepatic oval cells, including enlargement of the oval cells, a decrease in nucleus to cytoplasm ratio, and a 50% increase in the number of binucleated cells. Western blot results showed that 0.75 mmol/L sodium butyrate markedly raised the expression of albumin.

CONCLUSIONSodium butyrate, a differentiation promoting agent, can induce rat hepatic oval cells (liver progenitor cells) to differentiate into mature hepatocytes in vitro.

Animals ; Butyrates ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hepatocytes ; cytology ; Liver ; cytology ; Rats ; Stem Cells ; cytology

OBJECTIVESElevated tissue inhibitor of metalloproteinase-1 (TIMP-1) expression contributes to excess extracellular matrix in liver fibrosis. This study was designed to construct two recombinant adeno-associated viruses (AAV) carrying antisense RNA and small interfering RNA (siRNA) of TIMP-1 (rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo), and then to compare the suppression of TIMP-1 gene expression on rat hepatic stellate cell (HSC)-T6 cells infected by these two types of viruses in vitro.

METHODSAntisense RNA amplified by rat HSC-T6 and U6 promoter followed by the annealing siRNA were cloned into the AAV vector (pdl6-95/neo) and packed in 293 cells to construct the recombinants rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo. Rat HSC-T6 cells were infected with these recombinant AAVs and selected by using G418, and real-time PCR after reverse transcription and Western blot were performed to detect the transcription and expression level of TIMP-1 gene in these cells.

RESULTSThe results of PCR, restrictive enzyme digestion and gene sequencing confirmed that the pdl6-95/ANTI-TIMP-1/neo and pdl6-95/siRNA-TIMP-1/neo had been reconstructed successfully. After they had been packed in 293 cells to form rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo, they were used to infect HSC-T6. Thirty days after the infection, the transcription level of TIMP-1 in HSC-T6 cells infected by rAAV/siRNA-TIMP-1/neo decreased dramatically compared with the mock control and normal HSC-T6 cells (P less than 0.01), and the expression level of TIMP-1 gene in HSC-T6 cells decreased significantly (60%), while the transcription and expression level of TIMP-1 in HSC-T6 cells infected by rAAV/ANTI-TIMP-1/neo had no significant difference with mock control and normal HSC-T6 cells (P more than 0.05).

CONCLUSIONRNA interference can exert a suppression of TIMP-1 gene in rat HSC, and when this function combines with AAV infection, it can suppress the specific gene expression for a long time by chromosomal integration.

Animals ; Cells, Cultured ; Dependovirus ; genetics ; Genetic Vectors ; Hepatic Stellate Cells ; metabolism ; RNA, Antisense ; genetics ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo investigate the polymorphism of DXS6801, DXS6809, DXS7423, DXS7424, DXS9902 five loci in Hebei Han population.

METHODSThe PCR products were detected by the polyacrylamide gel electrophresis and DNA sequencing analysis.

RESULTSAmong 114 irrelative males and 118 irrelative females from Hebei Han population, 31 alleles were found in the 5 loci. One hundred and one haplotypes of the male were detected and the haplotype diversity reached 0.9975.

CONCLUSIONThe five loci are relatively abundant in polymorphic information for identification and paternity test. And the obtained data of Hebei Han population can be applied to the X-short tandem repeat genetic data bank.

Alleles ; Base Sequence ; China ; Chromosomes, Human, X ; genetics ; Female ; Gene Frequency ; Haplotypes ; Humans ; Male ; Microsatellite Repeats ; genetics ; Molecular Sequence Data ; Polymorphism, Genetic ; genetics

OBJECTIVETo evaluate the clinical efficacy of ultrasound-guided lauromacrogol sclerotherapy for benign thyroid cysts and analyze the factors affecting the efficacy.

METHODSUltrasound-guided lauromacrogol sclerotherapy was performed in 97 patients with a total of 99 benign thyroid cysts. The changes in cystic volume and other thyroid parameters were evaluated at 1, 3, 6, and 12 months after sclerotherapy. According to changes in the cystic volume, the efficacy of sclerotherapy was defined as therapeutic failure (with a volume reduction <50%), treatment success (volume reduction ≥50%) and cure (volume reduction ≥90%). The factors of affecting the efficacy of sclerotherapy was analyzed using COX regression.

RESULTSThe mean cystic volume at 1, 3, 6 and 12 months after sclerotherapy were reduced from the baseline volume of 12.08∓11.56 cmto 5.63∓8.51 cm, 5.96∓8.42 cm, 3.80∓5.50 cmand 2.85∓3.98 cm, respectively, with an average cystic volume reduction rate of (70.02∓33.72)%. Therapeutic success was achieved 82 of the 99 cysts (82.83%) and cure was achieved 63cysts (63.64%) at 12 months after the procedure. A second sclerotherapy was performed for 13 cysts which did not show a volume reduction at 1-3 months after the initial procedure. A disease course of over 12 months was an independent risk factor for a second sclerotherapy (23.7% [9/38] vs 6.6% [4/61], OR=4.473 [1.238-16.169], P=0.022). The efficacy of sclerotherapy was related to cystic cavity separation, cystic fluid viscosity, cystic/solid ratio and cystic wall thickness. COX regression analysis revealed that cystic cavity separation (HR=2.25, 95%CI: 1.19-4.25) and cystic fluid viscosity (HR=2.02, 95%CI: 1.19-3.43) were the major factors affecting the treatment efficacy.

CONCLUSIONUltrasound-guided lauromacrogol sclerotherapy is effective and safe for treatment of benign thyroid cysts, and the maximal treatment effect can be achieved at 6 months after sclerotherapy and in cases of uncomplicated cysts with non-viscous cystic fluid, no solid cystic cavity separation and a disease course of less than 12 months.

OBJECTIVETo evaluate visual inspection with Lugol's iodine (VILI) in cervical cancer screening program and to provide evidence for designing a cervical cancer screening algorithm in high risk areas of existing low-resource settings to reduce the incidence and mortality of cervical cancer.

METHODSWomen in Yangcheng county, Shanxi province were screened with VILI, colposcopy, liquid-based cytology test and human papilloma virus (HPV) DNA test. The efficacy of different screening tests was compared by Youden's index based on the pathology as the gold standard.

RESULTSIn the population being screened, the mean age was 40.80 +/- 10.75 years old. Based on pathological findings, 4.35% (32/735) of the subjects had >or= CIN (cervical intraepithelial neoplasia) II. The sensitivity and specificity for the VILI test (>or= positive) were 53.13 and 82.19, while 56.25 and 79.09 were for colposcopy (>or= low grade dysplasia) respectively. Comparing by the Youden's indexs, there was no statistically significant difference (P > 0.05) between VILI and colposcopy. However, statistical significant difference (P < 0.05) was found between VILI and liquid-based cytology test and HPV DNA test. In addition, there was no statistically significant difference (P > 0.05) found between the experienced doctors and the newly-trained doctors working in the field station.

CONCLUSIONWith low sensitivity when using microscope but low cost of equipments, VILI can be one of the primary screening tests in China's rural area with low-resource settings if the screening frequency is to be increased.

Adult ; Cervical Intraepithelial Neoplasia ; diagnosis ; epidemiology ; China ; epidemiology ; Early Detection of Cancer ; economics ; methods ; Female ; Health Resources ; Humans ; Incidence ; Iodides ; Program Evaluation ; Rural Health ; Sensitivity and Specificity ; Socioeconomic Factors ; Uterine Cervical Neoplasms ; diagnosis ; epidemiology

OBJECTIVETo investigate the distribution of the polymorphism of the Y-chromosomal loci DYS438, DYS439, GATA A7.1 and GATA A7.2 among Han population in Hebei province.

METHODSWith the use of PCR followed by polyacrylamide gel electrophoresis and silver staining, the allele frequencies of these loci in 164 unrelated men of Han population were investigated.

RESULTSFour, five, five, four alleles were observed at the loci DYS438, DYS439, GATA A7.1 and GATA A7.2 respectively; the frequencies of these alleles ranged from 0.0359 to 0.6587, from 0.0179 to 0.4107, from 0.0122 to 0.4146 and from 0.0476 to 0.5238 respectively; the probability discrimination of these loci were 0.5121, 0.6811, 0.6679 and 0.6327 respectively. Seventy different haplotypes were found at these loci. Thirty-six different haplotypes appeared only once. The power of discrimination of these four loci was 0.9480.

CONCLUSIONThe results demonstrate that these loci(DYS438, DYS439, GATA A7.1 and GATA A7.2) are good genetic markers with high determination power and can be applied to individual identification, especially in paternity test and the detection of mixed samples.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Chromosomes, Human, Y ; Gene Frequency ; Genetic Markers ; Genetics, Population ; Genotype ; Haplotypes ; Humans ; Male ; Paternity ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

OBJECTIVEHigh short-term successful rate was reported for catheter ablation in patients with paroxysmal atrial fibrillation (AF), we analyzed the long-term outcome (success rate, anticoagulation therapy and embolism event, anti-arrhythmic therapy and death post procedure) of catheter ablation for paroxysmal AF in this study.

METHODSFrom January 2000 to December 2004, 106 consecutive patients with drug-refractory paroxysmal AF underwent catheter ablation and were followed-up for (60.7 + or - 11.8) months. Segmental pulmonary vein isolation (SPVI) was routinely performed by radiofrequency energy under the guidance of circular mapping catheter. The patients were followed up with 24 h-holter, ECG, telephone or letter. Data on recurrence of AF, the anticoagulation medication and the incidence of embolism, anti-arrhythmic therapy were obtained.

RESULTSThere were 9 patients lost to follow up. In the remaining 97 patients [65 males, (54.8 + or - 11.2) years old], 3 cases died from cancer, sinus rhythm was maintained in 68 patients (Group S, 72.3%) and AF recurrence evidenced in 26 patients (Group R, 27.7%). In Group S, 56 patients (82.4%) discontinued anticoagulation medication, and 12 patients continued to take aspirin. There was no embolism event in Group S during follow-up. In Group R, 1 patient continued to take warfarin; 11 patients continued to take aspirin and 2 patients suffered from cerebral embolism. Anticoagulation medication was discontinued in 14 patients (53.8%) and 1 patient suffered form cerebral embolism. The incidence of embolism event in Group R is significantly higher than in Group S (P < 0.01). More patients discontinued anti-arrhythmic medication in Group S than in Group R (80.9% vs. 56.0%, P < 0.05).

CONCLUSIONCatheter ablation is associated with satisfactory long-term success rate, reduced anti-arrhythmia medication, improved quality of life in patients with paroxysmal AF.

Adult ; Aged ; Atrial Fibrillation ; therapy ; Catheter Ablation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies