Objective To investigate the therapeutic effect of CTLA4Ig gene on experimental autoimmune thyroiditis (EAT) by the use of portable synthetic costimulatory molecules of cytotoxic T lymphocyte antigen 4 (CTLA-4) antagonist (CTLA4Ig) eukaryotic expression vector.Methods Thirty C57BL/6J female mice were divided into three groups,named EAT model group (EAT,n =10),CTLA4Ig-treatment group (CTLA4Ig-EAT,n =10) and control group(n =10).At 28 day after first immunization,plasmids mixture with pCI or pCI/CTLA4Ig were injected into thyroid tissues of EAT and CTLA4Ig-EAT by surgery,respectively.Serum,thyroid tissues and spleens were collected as samples.Thyroid autoantibody and expression of interleukin (Th)1,Th2 related cytokinesby were measured by real-time quantitative PCR and ELISA.Results Compared with EAT group,the expression of CTLA-4 in thyroid of CTLA4Ig-EAT group was elevated double folds (P =0.038),and the expression of Th1 cytokine interferon γ and intercellular adhesion molecule-1 decreased significantly (P =0.016,0.042).Meanwhile,Th2 cytokine IL-4 was increased after CTLA4Ig treatment (P =0.044).The same changes were seen in spleen tissues and serum.There was no significant difference in terms of TPOAb between EAT and treated group.Conclusion Local thyroid injection of CTLA4Ig gene shows the therapeutic effect to same degree on EAT through adjusting the underlying Th1/Th2 imbalance.

Objective: To analyze the VP1 gene characteristics of Coxsackievirus A16 (CV-A16) isolated in(hand, foot and mouth disease, HFMD) patients of Jinan 2017. Methods: The samples collected from patients with HFMD in Jinan city in 2017 were analyzed and some of the CV-A16 nucleic acid positive samples were choosen with simple random sampling method and pretreated to inoculate RD cells. The whole VP1 gene sequences of CV-A16 stains which had obvious cytopathic effect in RD cells were amplified and sequenced. MEGA5.2 software was used to constructed phylogenetic tree and Lasergene software was used to analysis the homology consistency of nucleotide and amino acid of VP1 gene. Results: All the 10 CV-A16 strains isolated from Jinan were located 3 clusters belonged to B1b genotypes, with a nucleotide similarity of 94.9%~100% and deduced amino acid similarity of 99.3%~100%. The Jinan CV-A16 strains were nearest related with Shanghai strains KX871333(nucleotide similarity: 99.7%), Henan strains KM260134(99.0%), Jiangsu strains KP751580、KR138327(98.7%) and Guangdong strains MH004016(98.5%). Compared with some reference strains from our country, amino acid mutation were identified at positions 14, 59, and 145 in VP1 proteins of Jinan CV-A16 strains. Conclusions CV-A16 strains B1b genotypes were the strains prevailed in Jinan city, and 3 amino acid substitutions in VP1 proteins were happened in CV-A16 strains isolated from Jinan.

OBJECTIVETo evaluate the correlation between the mean of time blood flow velocity (TVmean) of feeding artery around hepatocellular carcinoma (HCC) and the microvessel density (MVD) in relation to tumor cell differentiation, and to evaluate the usefulness of TVmean as a noninvasive preoperative marker of biologic characteristic of HCC.

METHODSTo measure TVmean of feeding arteries around the HCC in 45 patients before operation and TVmean of minute arteries in hepatic tissue in 45 normal subjects by ultrasonographic examination. Tumor cell differentiation grade was determined by routine histopathological staining. Tumor MVD was measured by immunohistochemical staining with monoclonal antibodies against F VIII RAg of excised HCC. Correlationship between TVmean of feeding arteries and MVD of HCC was studied.

RESULTSA linear correlation between TVmean of feeding arteries around the tumors of HCC and MVD of HCC tissue (r = 0.794, y = 18.2764 + 1.5544x) was obtained. With TVmean > or = 21.3 cm/s, the positive and negative predictive values of poorly differentiated HCC were 97.4% and 87.5%, respectively. With TVmean < 21.3 cm/s, those of well differentiated HCC were 88.5% and 97.3%, respectively. The degree of reliability of calculating HCC cell differentiation according to TVmean was 89.5%.

CONCLUSIONTumor growth and cellular differentiation, the two major factors affecting prognosis of HCC, may be evaluated by TVmean of feeding arteries around the tumor. It offers a useful preoperative information for predicting prognosis of HCC patients.

Adult ; Aged ; Arterioles ; pathology ; Blood Flow Velocity ; Capillaries ; pathology ; Carcinoma, Hepatocellular ; blood supply ; pathology ; physiopathology ; Female ; Humans ; Liver Neoplasms ; blood supply ; pathology ; physiopathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology

Objective: To analyse the genomic characteristics of Novirus(NoV) GII.4 genotype JN010 strain isolated form Jinan in 2017. Methods: Seven pairs of primers were designed and used to amplify the JN010 genome. Sequence analyses, alignment and phylogenetic trees of ORF1 and ORF2 genes were performed using the software Lasergene7.1 and MEGA5.2. At the same time the major protein VP1 amino acid mutations were analyzed. Results: The 7 516 bp complete genome sequence of JN010 strain was obtained, the most mutation sites were located in P2 subdomain of VP1. Two substitutions I293N and H373N of VP1 were locate neighboring epitope A, and R297H mutation happened within epitope A and the site A that binding with histo-blood group antigens(HBGAs). The JN010 strain was GII.Pe/GII.4 genotype and genetically closest to the strains found in Osaka of Japan(GenBank accession number LC066046) and the strains in Zhongshan city of Guangdong (GenBank accession number KY407064) respectively according to ORF1 and ORF2 gene homologous and phylogenetic analysis. Conclusions The NoV GII.4 variant strain JN010 has occurred mutations in the key site of the epitope A and site A that bind with HBGAs, and maybe affect its antigenicity and interaction with HBGAs.

Regeneration of periodontal tissue is the most promising method for restoring periodontal structures. To find a suitable bioactive three-dimensional scaffold promoting cell proliferation and differentiation is critical in periodontal tissue engineering. The objective of this study was to evaluate the biocompatibility of a novel porcine acellular dermal matrix as periodontal tissue scaffolds both in vitro and in vivo. The scaffolds in this study were purified porcine acellular dermal matrix (PADM) and hydroxyapatite-treated PADM (HA-PADM). The biodegradation patterns of the scaffolds were evaluated in vitro. The biocompatibility of the scaffolds in vivo was assessed by implanting them into the sacrospinal muscle of 20 New Zealand white rabbits. The hPDL cells were cultured with PADM or HA-PADM scaffolds for 3, 7, 14, 21 and 28 days. Cell viability assay, scanning electron microscopy (SEM), hematoxylin and eosin (H&E) staining, immunohistochemistry and confocal microscopy were used to evaluate the biocompatibility of the scaffolds. In vitro, both PADM and HA-PADM scaffolds displayed appropriate biodegradation pattern, and also, demonstrated favorable tissue compatibility without tissue necrosis, fibrosis and other abnormal response. The absorbance readings of the WST-1 assay were increased with the time course, suggesting the cell proliferation in the scaffolds. The hPDL cells attaching, spreading and morphology on the surface of the scaffold were visualized by SEM, H&E staining, immnuohistochemistry and confocal microscopy, demonstrated that hPDL cells were able to grow into the HA-PADM scaffolds and the amount of cells were growing up in the course of time. This study proved that HA-PADM scaffold had good biocompatibility in animals in vivo and appropriate biodegrading characteristics in vitro. The hPDL cells were able to proliferate and migrate into the scaffold. These observations may suggest that HA-PADM scaffold is a potential cell carrier for periodontal tissue regeneration.
Absorbable Implants ; Acellular Dermis ; Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Culture Techniques ; Cell Movement ; physiology ; Cell Proliferation ; Cell Shape ; physiology ; Cell Survival ; physiology ; Durapatite ; chemistry ; Fibrosis ; Humans ; Microscopy, Electron, Scanning ; Muscle, Skeletal ; surgery ; Necrosis ; Periodontal Ligament ; cytology ; surgery ; Rabbits ; Regeneration ; physiology ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

We constructed a Brain-computer interface-based mental speller which realizes user-computer interaction. The feature signals of user's intention are embedded in spontaneous EEG background. Single-trial feature estimation should be used on this online occasion instead of the grand average usually used in cognitive or clinical experiments. To demonstrate this technique beyond laboratories, fewer EEG recording channels are preferred. A unique paradigm, which is called imitating-natural-reading, was exploited to induce visual evoked potentials. We explored the single-trial estimation of VEP recorded in single channel using support vector machine on three subjects, and obtained satisfactory data, the classification accuracy being 92.1%, 94.1% and 91.5%, respectively. These results put forward a significant step fowards the ultimate realization of our mental speller.
Adult ; Algorithms ; Brain ; physiology ; Electroencephalography ; Evoked Potentials, Visual ; physiology ; Humans ; Male ; Signal Processing, Computer-Assisted

PURPOSE: The purpose of this study was to explore the potential early diagnostic value of serum microRNA-381(miRNA-381) in patients with gastric cancer (GC). MATERIALS AND METHODS: Patients with advanced gastric cancer (AGC) and early gastric cancer (EGC), as well as healthy individuals, were enrolled in this study. Expression of miRNA-381 in serum was detected using real-time quantitative PCR. Electrochemiluminescence analysis was used to investigate the expression of classic tumor markers, including carbohydrate antigen (CA) 199, CA724, and carcinoembryonic antigen. Finally, receiver operating characteristic curve and Kaplan-Meier analysis were used to determine the value of miRNA-381 in clinical diagnosis of GC. RESULTS: miRNA-381 was differentially expressed among the study groups. AUC analysis showed that the sensitivity and specificity of serum miRNA-381 in the diagnosis of GC were superior to those of other tumor markers. Furthermore, low levels of miRNA-381 expression were positively correlated with lymph node metastasis and AGC. Finally, Kaplan-Meier survival analysis showed that down-regulation of miRNA-381 was associated with lymph node metastasis and the development of GC. CONCLUSION: miRNA381, which was down-regulated in GC, might be a novel early diagnosis marker for patients with GC.
Area Under Curve ; Biomarkers, Tumor ; Carcinoembryonic Antigen ; Diagnosis ; Down-Regulation ; Early Diagnosis ; Humans ; Kaplan-Meier Estimate ; Lymph Nodes ; Neoplasm Metastasis ; Polymerase Chain Reaction ; ROC Curve ; Sensitivity and Specificity ; Stomach Neoplasms

OBJECTIVE: To investigate the mechanism of inflflammatory-mediated toll-like receptor 4 (TLR4)-p38 mitogen-activated protein kinase (p38 MAPK) pathway in Kupffer cells (KCs) of non-alcoholic steatohepatitis (NASH) rats and the intervention effect of soothing Gan (Liver) and invigorating Pi (Spleen) recipes on this pathway. METHODS: After 1 week of acclimatization, 120 Sprague-Dawley male rats were randomly divided into 8 groups using a random number table (n=15 per group): normal group, model group, low-dose Chaihu Shugan Powder (, CHSG) group (3.2 g/kg), high-dose CHSG group (9.6 g/kg), low-dose Shenling Baizhu Powder (, SLBZ) group (10 g/kg), high-dose SLBZ (30 g/kg) group, and low- and highdose integrated recipe (L-IR, H-IR) groups. All rats in the model and treatment groups were fed with a high-fat diet (HFD). The treatments were administrated by gastrogavage once daily and lasted for 26 weeks. The liver tissues were detected with hematoxylin-eosin (HE) and oil red O staining. Levels of liver lipids, serum lipids and transaminases were measured. KCs were isolated from the livers of rats to evaluate the mRNA expressions of TLR4 and p38 MAPK by real-time flfluorescence quantitative polymerase chain reaction, and proteins expressions of TLR4, p-p38 MAPK and p38 MAPK by Western blot. Levels of inflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin (IL)-1 and IL-6 in KCs were measured by enzyme-linked immunosorbent assay. RESULTS: After 26 weeks of HFD feeding, HE and oil red O staining showed that the NASH model rats successfully reproduced typical pathogenesis and histopathological features. Compared with the normal group, the model group exhibited significant increases in body weight, liver weight, liver index, serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol, and aspartate aminotransferase as well as TC and TG levels in liver tissues, and significant decrease in serum level of high-density lipoprotein cholesterol (Plt;0.05 or Plt;0.01), while those indices were significantly ameliorated in the H-IR group (Plt;0.05 or Plt;0.01). Higher levels of TNF-α, IL-1 and IL-6 in KCs were observed in the model group compared with the normal group (Plt;0.01). Significant decreases in TNF-α, IL-1 and IL-6 were observed in the H-SLBZ, H-IR and L-IR groups compared with the model group (Plt;0.05 or Plt;0.01). The mRNA expressions of TLR4 and p38 MAPK and protein expressions of TLR4, p38 MAPK and p-p38 MAPK in KCs in the model group were significantly higher than the normal group (Plt;0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group (Plt;0.05 or Plt;0.01). CONCLUSION Inflflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38MAPK signaling pathway in KCs for the anti-inflflammatory effect of soothing Gan and invigorating Pi recipes.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Kupffer Cells ; drug effects ; physiology ; MAP Kinase Signaling System ; drug effects ; Male ; Medicine, Chinese Traditional ; Non-alcoholic Fatty Liver Disease ; drug therapy ; physiopathology ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; physiology ; p38 Mitogen-Activated Protein Kinases ; physiology

Acute Disease ; Adult ; Humans ; Iliac Artery ; Lower Extremity ; blood supply ; Male ; Nephrotic Syndrome ; complications ; Thrombosis ; etiology