The aim of the present study was to work on the efficiency of differential diagnosis of native hapten-gel diffusion assay (NH-GD) on the background of vaccination with S2 or Rev.1.The conditions of NH-GD assay was firstly optimized,its sensitivity,specificity,repeatability and ability of differential diagnosis were determined respectively,and its test result was compared with that of fluorescence polarization assay (FPA).The results showed NH-GD assay with good specificity and repeatability could differentiate Brucella-infected antibody from vaccinated antibody after vaccination with S2 or 122 days after vaccination with Rev.1.And the result of NH-GD assay was highly consistent with that of FPA,which was simple to operate and needed a few simple equipment.Therefore,NH-GD assay was a good method for sheep brucellosis surveillance in China and especially suitable for application in grass-roots areas.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Objective To explore the effect of levetiracetam(LEV)on cognitive function and quality of life of patients with partial seizure.Methods There were two phase for this study.Phase one was a randomized,double-blind,placebo-controlled study for 16 weeks.Phase two was an open label period for 24 weeks.Medicated patients underwent test on the cognitive function and Health Related Quality of Life (HRQOL)at the baseline respectively at the end of 16 weeks and at the end of 40 weeks.Results(1) Compared the result at the end of 16 weeks with the baseline,LEV group used less time to do Wisconsin card sorting test(WCST,reduce scores -110.3),delayed logical memory were improved(reduce scores 3.4,P

The prevalence of thyroid cancer has shown an upward trend in China in recent years. Advances in thyroid ultrasound and fine needle puncture cytology have improved the accuracy of the preoperative diagnosis of thyroid cancer. Also,the application of endoscopy-assisted techniques and intraoperative nerve monitoring technology and the further understanding of thyroid lymph node metastasis have made the thyroid surgeries safer and less invasive. This article summarizes the recent advances in the surgical therapy of thyroid cancer.
Carcinoma, Papillary ; surgery ; Humans ; Iodine Radioisotopes ; therapeutic use ; Molecular Targeted Therapy ; Neoadjuvant Therapy ; Thyroid Neoplasms ; surgery ; therapy

Objective To investigate the expression of vascular endothelial growth factor(VEGF)in synovium of rats with adjuvant arthritis and the relationship between the histopathologic score and the expression of VEGF.Methods Adjuvant arthritis was established in Wistar rats by inoculating complete Freund's adjuvant(CFA). We calculated the arthropathologic score and the expression of VEGF mRNA and protein at different stages after CFA inoculation.Results In model group the arthropathologic score and expression of VEGF protein in synovium increased significantly all the time (P

To investigate the differentiation of bone mesenchymal stem cells(BMSCs) into chon-drocytes by miRNA-206 and its mechanism in osteoarthritis(OA). Methods From January, 2017 to July, 2018, rat BMSCs were isolated, and their CD90 and CD45 were detected by flow cytometry. Transfection of miRNA-206 or miRNA-206 inhibitors into BMSCs using lentiviral vectors, dexamethasone induction for 14 d, then use alician blue staining and type II collagen immunostaining to detect chondrogenic differentiation. MTT assay was used to detect the proliferation of mesenchymal stem cells. Western blot analysis was used to detect the Aggrecan, Col II, Sox9 and Runx2 markers in chondroblast cells. The expression level of the marker gene of Sox9 mRNA in chondroblasts were detected by RT-PCR.OA rat models were treated with lentiviral vectors transfected with miRNA-206 or miRNA-206 inhibitors, and Aggrecan, Col II, Sox9, Runx2 which were the markers of chondrogenesis were detected by Western blot. Results The purity of isolated BMSCs was (80.7±3.9)%. BMSCs transfected with miRNA-206 could promote cell proliferation and increase chondrogenic differentiation. Western blot results showed that the expression of Aggre-can, Col II and Sox9 was increased in the miRNA-206 transfection group, and the expression of Runx2 was down-regulate. Meanwhile, RT-PCR results showed that miRNA-206 can up-regulate the expression of the chondroblast marker gene Sox9 mRNA in BMSCs.Compared with the OA group, miRNA-206 could increase the expression of Aggre-can, Col II and Sox9 signaling proteins in cartilage tissue (P＜0.05), and down-regulate the expression level of Runx2 (P＜0.05). Conclusion The miRNA-206 can positively regulate the differentiation of BMSCs into chondrocytes, increase the ability of cell proliferation, up-regulate the expression of Aggrecan, Col II and Sox9, and down-regulate Runx2.The miRNA-206 increase chondrogenic capacity in rat models of osteoarthritis.

Objective To investigate the polymorphism of human cytomegalovirus (HCMV) UL144 gene of the low passage clinical isolates in Guangzhou and explore the role of UL144 gene in HCMV pathogenicity. Methods The clinical isolates of HCMV were obtained from the urine sample collected from those infants with intra-uterus HCMV infection in Guangzhou. The virus genome DNA was extracted. According to the genome sequence of Toledo, primers for UL144 gene were designed and used to amplify the complete open reading frames (ORF) of the UL144 gene in our 3 different clinical isolates. These ORFs of the UL144 gene were cloned into pMD18-T vector and their sequences were confirmed by sequencing. Bioinformatics methods were used subsequently to analyze the polymorphisms of these genes in different stains. Results Three HCMV low passage clinical isolates were successfully isolated, named D2, D3 and D52. As shown by PCR, all of these three strains contained UL144 ORF region. Three complete ORFs were amplified in total and their sequences were submitted to GenBank (Accession No.: DQ180368, DQ180382 and DQ180355). In D2, D3 and D52 isolates, their UL144 ORFs consisted of 531 nucleotides. DNA sequences were quite conservative,all variability were base substitution, and the amino acid sequences were high conservative, the rate of amino acid variability was 1.1%. There were no additional or deleted sites of posttranslational modification of UL144 protein in all clinical isolates. There were some differences in the secondary structure among different isolates. The isoelectric point of UL144 protein of all clinical isolates was 8.97. Conclusions All DNA and deduced amino acid sequences of UL144 gene share great similarity among Guangzhou HCMV clinical strains regardless of their polymorphism. It implies that maybe UL144 gene plays an important role in congenital infection.

OBJECTIVETo establish a digital model allowing three-dimensional visualization of the structures involved in the anterior cervical segment approach.

METHODSBased on the imaging data obtained from CT angiography (CTA), magnetic resonance myelography (MRM) and continuous magnetic resonance imaging (MRI) of a healthy volunteer (scanning from the center of the head to the inferior border of the T3 level), image segmentation and reconstruction for the skeleton, arteries, veins, and spinal cord was conducted semi-automatically using the Mimics software according to the different thresholds of the tissues. The cervical plexus, brachial plexus and muscles of the neck were reconstructed with the Nerves pipe editor and the Med CAD module to establishing the three-dimensional model for displaying the structures involved in the anterior cervical segment approach.

RESULTSA three-dimensional digital model of the structures involved in the anterior cervical segment anterior approach was established, which allowed the display of anatomical relations of the skeletal structure, aorta, superior vena cava, thyroid gland, hyoid bone, laryngeal cartilages, trachea, lung, 12 neck muscle groups, as well as the spinal cord, spinal nerves, cervical plexus, brachial plexus, and intervertebral disk of the neck.

CONCLUSIONThe three-dimensional model established can allow the visualization of the important structures for the anterior cervical segment approach, and provides a medical teaching platform for anatomy and surgical training.

Adult ; Cervical Vertebrae ; anatomy & histology ; diagnostic imaging ; surgery ; Computer-Aided Design ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Models, Anatomic ; Neck ; anatomy & histology ; Tomography, Spiral Computed ; methods

Objective The aim of this study was to investigate the effect of TSP-1 gene on angiogenesis in human osteosar-coma and its mechanism of action. Methods MG-63 cells were transfected with constructing pBPLV-shRNA-TSP-1 vector and pBPLV-TSP-1 expression vector. Cell viability was measured by CCK8,and its invasive ability was measured by Transwell assay. The expression of CD36 in intracells was detected by immunofluorescence. The expression levels of TSP-1,CD36,p38MAPK,VEGF, VEGFR-1,EGF and PDGF were detected in MG-63 cells by qRT-PCR and Western blot. Results The cell viability and inva-sion ability were significantly increased after transfected pBPLV-TSP-1 vector compared with the empty vector group(P<0. 05), and significantly decreased after transfected pBPLV-shRNA-TSP-1 vector( P<0. 05). The expression of TSP-1,EGF,P38, PDGF,VEGF and VEGFR -1 at mRNA and protein levels was significantly increased after transfection pBPLV -TSP -1 ( P <0. 05),and significantly decreased after transfection pBPLV-shRNA-TSP-1 vector(P<0. 05). Conclusion TSP-1 gene can promote the proliferation and invasion of MG-63 cells,and promote the formation of human osteosarcoma,indicating its mechanism related to the increase of growth factors EGF,VEGF,PDGF and activation of P38-MAPK pathway.

OBJECTIVESTo investigate S gene mutations in HBsAg/HBsAb double positive chronic hepatitis B patients.

METHODSHBV S gene from 8 patients (Group A) with HBsAg (+)/HBsAb (+) and 9 patients (Group B) with HBsAg (+)/HBsAb (-)was amplified by polymerase chain reaction (PCR) and sequencing. Both the distribution of genotype and serotype and the rate of MHR region were compared by Fisher's exact test. The mutation rate of both the DNA level and amino acid level was compared by t test.

RESULTSNo significant difference in distribution of genotypes was found between the two groups (P=0.153). In group A, 2 were genotype B, 6 were genotype C; In group B, 6 were genotype B, 3 were genotype C. No significant difference in distribution of serotypes was found between the two groups, either (P=0.218). In group A, 2 were adw, 5 were adr, 1 was ayr; In group B, 6 were adw, 3 were adr. The mutation rate of Pre-S1 region at both the DNA level (2.29% vs 1.80%, t=2.66, P more than 0.05) and the amino acid level (2.66% vs 1.59%, t=1.39, P>0.05) was not significantly different between these two groups; the mutation rate of Pre-S2 region in group A patients was significantly higher than that in group B at the DNA level (1.74% vs 0.91%, t=4.68, P<0.01), but not higher at the amino acid level (3.18% vs 2.05%, t=1.85, P>0.05), the mutation rate of S region in group A patients was significantly higher than that in group B at both the DNA level (2.13% vs 0.81%, t=6.00, P<0.01) and the amino acid level (4.37% vs 1.52%, t=5.32, P<0.01). Amino acid substitutions were found both within and beyond the MHR region. The rate of "a" determinant mutations in these two groups was also found to be significantly different (P<0.05).

CONCLUSIONHigher HBV S gene mutation rate exists in HBsAg/HBsAb double positive patients than that in HBsAg (+)/HBsAb (-) patients.

Adult ; Aged ; Base Sequence ; DNA, Viral ; blood ; genetics ; Female ; Genes, Viral ; Genetic Variation ; Genotype ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Young Adult