A set of primers amplified the VP1 gene of foot-and-mouth disease vims (FMDV) was designed and synthesized. A reverse transcription-polymerase chain reaction (RT-PCR) technique detected the RNA of FMDV was established after selecting the best purification method, reagents and reaction conditions. Samples of fresh milk, lymph node, spinal cord, vesicular skin, milk powder, cotton swab, mouse and meat in daughter-house were detected by RT-PCR, positive rates were41.4% (24/58), 13.33% (2/15), 20% (1/5), 100% (1/1), 100% (1/1), 37.5% (12/32), 100% (2/2) and 10% - 70%, respectively. However, positive rate of cockroach detected by RT-PCR was 0. The results showed that the established FMDV RT-PCR technique provided a more sensitive, specific and reliable method for diagnosis and epizootic study of the foot-and-mouth disease.

Animals ; Behavior, Animal ; Reproduction ; physiology ; Social Behavior ; Tupaiidae ; physiology

OBJECTIVETo investigate the effects of catecholamine hormone on the blood and brain of heroin addicts.

METHODSRats were divided into three groups and treated with the glucose (control group), the heroin (im) (heroin group), and the combination of the intramuscular injection of reserpine and heroin (reserpine group). Changes in the levels of the dopamine (DA), cAMP, and cGMP were detected by the radioimmunoassay (RIA) method in the blood and brain tissue.

RESULTSNo significant withdrawal symptoms were observed in the reserpine group. Compared with the control and heroin groups, the blood cAMP levels were increased by 35.36% and 15.53% in the reserpine group, respectively; the cAMP levels in the midbrain ventral tegmental area (VTA), prefrontal cortex (PFC), and hippocampus (Hipp) were increased by 24.08% & 8.53%, 15.66% & 8.13%, and 21.95% & 8.40%, respectively. While compared to the control and heroin groups, the DA levels of the PFC, Hipp, striatum, and nucleus accumbens (NAc) were significantly reduced in the reserpine group, decreasing by 74.09% & 82.86%, 81.06% & 82.23%, 91.62% & 86.55% and 84.35% & 90.63%, respectively. The concentrations of cGMP of the brain tissues in the reserpine group were lower than those in the control group. In addition, the neural electrophysiological testing showed that the electroencephalogram (EEG), electrocardiogram (ECG), and muscle spindle discharge diagram of rats in both the reserpine and heroin groups were apparently changed.

CONCLUSIONCatecholamine hormone plays an important role in heroin addiction.

Animals ; Brain ; drug effects ; metabolism ; Catecholamines ; physiology ; Cyclic AMP ; blood ; metabolism ; Cyclic GMP ; blood ; metabolism ; Dopamine ; blood ; metabolism ; Heroin Dependence ; metabolism ; physiopathology ; Male ; Rats ; Rats, Wistar

Objective To evaluate the feasibility of reconstructing horizontal periodontal bone defects by tissue engineering based on bone marrow stromal cells(BMSCs)as seed cells and enamel matrix proteins(EMPs)as growth factors. Methods Two healthy rhesus monkeys were selected, and BMSCs were isolated from iliac marrow and serial subcultivation was conducted. The cells of induced BMSCs at passage 3 were harvested and mixed with Bio-oss collagen. The models of horizontal periodontal bone defects were established surgically in each buccal side of the posterior teeth, and were divided into four groups (blank control group, material group, cells/material group and cells/material/EMPs group). The histological and Micro-CT observation were carried out 8 weeks later. Results In the blank control group, the defects were filled with fibrous connective tissue. There was newly-formed alveolar bone in the material group. In the cells/material group, periodontal regeneration could be observed, while the newly-formed cementum was irregular and less in quantity. In the cells/material/EMPs group, the amount of newly-formed alveolar bone was larger, and the newly-formed cementum was continuous and regular. Conclusion The tissue engineering technique of BMSCs as seed cells in combination with EMPs induction can significantly promote the regeneration of periodontal tissue.

Objective :To explore correlation between diabetic peripheral neuropathy (DPN) and cardiovascular disea-ses (CVD) in patients with type 2 diabetes mellitus (T2DM).Methods :A total of 375 T2DM patients ,who were treated in our community health service center from Jan 2014 to Jul 2016 ,were selected .Demographic characteris-tics ,systolic blood pressure (SBP) ,blood lipids ,blood glucose ,electromyogram and CVD were collected in all sub-jects using questionnaire .According to complicated CVD or not ,patients were divided into CVD group (n＝54) and non-CVD group (n＝321).Correlation among all indexes and CVD risk were analyzed .Multifactor Logistic regres-sion analysis was used to analyze independent risk factors for CVD risk .Results :Among the 375 T2DM patients ,54 cases (14. 4%) were complicated with CVD .Compared with non-CVD group ,there were significant rise in age≥65 years ,abnormal fasting blood glucose (FBG) , abnormal 2h postprandial blood glucose (2hPG) , abnormal glycosy-lated hemoglobin A1c (HbA1c) and SBP in CVD group , P＜0.05 or ＜0.01 .Spearman correlation analysis indica-ted that age ,FBG ,2hPG ,DPN ,SBP and HbA1c were significant positively correlated with CVD risk (r＝0.612～0.735 , P＝0.001 all).Multifactor Logistic regression analysis indicated that DPN , abnormal FBG and abnormal 2hPG were independent risk factors for CVD (OR＝1.248～2.023 , P＜0.05 all).Conclusion : Diabetic peripheral neuropathy is one of the independent risk factors for CVD in T2DM patients .Attention should be paid to preven-tion ,screening and early treatment of CVD in these patients in clinic .

OBJECTIVETo investigate the incidence and the gene mutation frequencies and patterns of β-thalassemia (β-Thal) in ethnic Han children in Chongqing city.

METHODA total of 1726 children were screened by using automatic hemocytic analyzer, cellulose acetate electrophoresis and fetal hemoglobin alkali denaturation test. Samples with mean corpuscular volume (MCV) < 80 fl, cell hemoglobin content (MCH) < 27 pg and hemoglobin A2 (HbA2) levels >3.3%, fetal hemoglobin (HbF) >2% for β-Thal screening indicators. The positive samples of screening indicators were detected and identified by PCR-reverse dot blot method for 18 common β-Thal mutations in Chinese populations, unknown mutations samples were subjected to DNA sequencing analysis of the β-globin gene.

RESULTTwenty-five cases of β-Thal carriers were observed from the 1726 samples, with 24 cases of β-Thal heterozygote and one case of double heterozygote. Therefore, the β-Thal carrier rate was 1.51%. After 1726 peripheral venous blood samples analyzed by hematological parameters, 164 positive cases of β-Thal screening indicators were found, with the positive rate being 9.50% (164/1726). A total of 6 different gene mutations were detected, the four most common mutations were as the following: CD41-42, IVS-II-654, CD17 and beta E. These four mutations as the major types in this area accounted for 88.00% of all the mutations. In addition, one rare mutation of 5 'UTR; + (43 -40) was found, and one case of the hemoglobin variant of Hb Zurich was reported in Chinese people for the first time.

CONCLUSIONChongqing is a high risk region of the β-Thal. Epidemiological Data from the research was useul for the genetic counseling and the prevention of β-Thal major.

Asian Continental Ancestry Group ; genetics ; Blood Chemical Analysis ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Counseling ; Hemoglobins ; analysis ; genetics ; Hemoglobins, Abnormal ; analysis ; genetics ; Heterozygote ; Humans ; Infant ; Male ; Mutation ; genetics ; Prevalence ; beta-Globins ; genetics ; beta-Thalassemia ; epidemiology ; ethnology ; genetics

MicroRNAs (miRNAs) have been demonstrated to play an important role in regulation of the immunoinflammatory response; however, the function of miRNAs in periodontal inflammation has not been investigated. The objective of this study was to explore the properties of miRNAs in periodontal inflammation by comparing miRNA profiles of inflamed and healthy gingival tissues. Gingival tissues were obtained from 10 periodontitis patients and 10 healthy subjects. After RNA extraction, miRNA profiles were analyzed by microarray, and expression levels of selected miRNAs were confirmed by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). Analyses using two computational methods, Targetscan and MicroRNA.org, were combined to identify common targets of these miRNAs. Finally, the individual miRNA expression levels of three toll-like receptor (TLR)-related miRNAs from inflamed and healthy gingival tissues were evaluated by RT-PCR. Ninety-one miRNAs were found to be upregulated and thirty-four downregulated over two-fold in inflamed gingival tissue compared with those in healthy gingival tissue. Twelve selected inflammatory-related miRNAs, hsa-miR-126*, hsa-miR-20a, hsa-miR-142-3p, hsa-miR-19a, hsa-let-7f, hsa-miR-203, hsa-miR-17, hsa-miR-223, hsa-miR-146b, hsa-miR-146a, hsa-miR-155, and hsa-miR-205 showed comparable expression levels by microarray and real-time quantitative RT-PCR analyses. In addition, the putative inflammation targets of these miRNAs were predicted, and three that were tested (hsa-miRNA-146a, hsa-miRNA-146b, and hsa-miRNA-155), showed significant differences between inflamed and healthy gingiva. This remarkable difference in miRNA profiles between periodontal diseased and healthy gingiva implicates a probable close relationship between miRNAs and periodontal inflammation. The data also suggest that the regulation of TLRs in periodontal inflammation may involve miRNA pathways.
Adult ; Case-Control Studies ; Chronic Periodontitis ; genetics ; metabolism ; Computational Biology ; methods ; Female ; Gene Expression Profiling ; Gingiva ; metabolism ; Humans ; Male ; MicroRNAs ; biosynthesis ; genetics ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA ; Toll-Like Receptors ; genetics

A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.
Gene Products, gag ; genetics ; Gene Products, pol ; genetics ; Human T-lymphotropic virus 1 ; genetics ; isolation & purification ; Humans ; Molecular Probes ; Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

OBJECTIVETo study the effect of perforator flaps of medial head of gastrocnemius muscle on repairing popliteal fossa scar wounds.

METHODSTen patients with scar in popliteal fossa hospitalized from January 2005 to January 2010 were repaired with perforator flaps of medial head of gastrocnemius muscle. The scar was resulted from burn in 8 patients, and from operation in 2. The duration of the scar was 3 months to 11 years, and area of the scar ranging from 6 cm × 3 cm to 10 cm × 6 cm. Ultrasonic Doppler was used to detect the musculocutaneous perforator vessel of the medial sural artery at the position 10 to 17 cm from the fold of the popliteal fossa and 2 to 5 cm from the posterior midline before surgery. Then flap transplantation surgery was performed. Donor site wounds with width less than 5 cm were sutured directly, and those wider than 5 cm were repaired with skin transplantation.

RESULTSAll the flaps survived. Flap size ranged from 7 cm × 5 cm to 12 cm × 7 cm. All patients were followed up for 3 to 30 months, and the flaps were found to have a good appearance. Patients could walk with heavy load without lameness. The function of knee joint of the affected limb was the same as that of the opposite limb. No obvious depression was observed in donor sites.

CONCLUSIONSThe perforator flaps of medial head of gastrocnemius muscle can be used to repair the popliteal fossa scar wound with satisfactory result.

Adolescent ; Adult ; Child ; Cicatrix ; surgery ; Female ; Humans ; Knee ; Male ; Muscle, Skeletal ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; Young Adult

OBJECTIVETo investigate the prognostic factors for patients with gastrointestinal stromal tumors (GIST).

METHODSFrom 2000 to 2003, clinical data of 41 cases with GIST were reviewed retrospectively. The clinicopathologic diagnosis was determined by immunochemistry. The relationships of the prognosis with mitotic counts, tumor size and location,range of tumor resection were analyzed.

RESULTSThe patients with GIST had pathological section of high expression in CD117, CD34 and vimentin (92.7%, 82.9%, 78%, respectively). Patients with tumor location in intestine, tumor size > 5 cm,mitotic counts > 5/50HPF, incomplete resection had poorer outcome, compared with those with tumor location in stomach and colon,tumor size < or = 5 cm,mitotic counts < or = 5/50HPF and complete resection (all P< 0.05).

CONCLUSIONComplete gross resection can improve prognosis for patients with GIST. Tumors with mitotic counts > or = 5/50HPF, tumor size more than 5 cm and tumor location in intestine are poor prognostic factors.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; diagnosis ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult