Serial analysis of gene expression (SAGE) is an effective method of determining gene expression profiles of tissues and organs under different conditions. In this paper, the detail protocol of SAGE was introduced and some modified procedure of SAGE was reviewed.
Gene Expression Profiling ; methods

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain ; drug effects ; pathology ; Disease Models, Animal ; Estrogens ; pharmacology ; Female ; Hypoxia-Ischemia, Brain ; drug therapy ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Wistar

Objective To evaluate the role and significance of Treg/Th17 cells imbalance in pathogenesis and recurrence mechanism of condyloma acuminatum (CA).Methods 52 patients with CA were selected as study group (CA group,27 with initial occurrence of CA,25 with recurrence of CA),30 healthy persons were as control group,flow cytometry was used to detect the proportion of Treg cells and Th17 cells in the peripheral blood,the expression level of Foxp3 mRNA and RORyt mRNA in peripheral blood mononuclear cells was determined by real-time quantitative polymerase chain reaction.Results The proportion of Treg cells and the expression level of Foxp3 mRNA in peripheral blood in CA group was higher than that in control group,recurrence CA group was higher than initial occurrence CA group,difference was significant(both P＜0.05);the proportion of Th17 cells and expression level of RORyt mRNA in CA group was significantly lower than that in control group,proportion of Th17 cells in recurrence CA group was lower than initial occurrence CA group,there was significant difference (both P＜0.05).The proporation of Treg/Th17 in CA group was higher than that in healthy controls(4.60[3.20,8.68] vs 1.39[1.05,2.05],P＜0.05),recurrence CA group was higher than initial occurrence CA group (8.19[4.21,10.81] vs 3.52 [2.47,4.85],P＜0.05).Conclusion There is an imbalance between Treg cells and Th17 cells in patients with CA,especially in patients with reccurrence of CA,the imbalance of Treg/Th17 cells may play an important role in the pathogenesis and recurrence mechanism of CA.

OBJECTIVE: To investigate the polymorphism of 11 canine STR loci. METHODS: A fluorescent multiplex system with 11 STR loci (PEZ1, PEZ2, PEZ3, PEZ5, PEZ6, PEZ8, PEZ12, FH2010, FH2054, FH2132 and FH2611) was constructed independently and performed to amplify 105 samples from dogs. The character of these loci was analyzed with the PCR data. RESULTS: The distributions of genotypes and allele frequencies of 11 STR loci were obtained. The total power of discrimination for the 11 loci in canine population was 0.9999999 and the cumulative probability of exclusion was 0.9330621. The observed heterozygosity and polymorphism information content (PIC) were 0.502 and 0.640, respectively. CONCLUSION Each of the eleven canine STR loci has a high genetic polymorphism and can be applied for the parentage testing and individual identification. The fluorescent multiplex system is a reliable method in forensic application.
Alleles ; Animals ; Dogs/genetics* ; Forensic Genetics ; Gene Frequency ; Genotype ; Microsatellite Repeats/genetics* ; Polymorphism, Genetic

Adult ; Anastomosis, Surgical ; Esophagus ; surgery ; Humans ; Male ; Stents ; Stomach ; surgery ; Tracheoesophageal Fistula ; surgery

Child ; Child, Preschool ; Electroencephalography ; Female ; Humans ; Male ; Monitoring, Physiologic ; Prognosis ; Status Epilepticus ; physiopathology ; therapy

OBJECTIVETo probe in the possible acting mechanism of Bizhongxiao Decoction (BZXD) for treatment of early active rheumatoid arthritis (RA) by way of observing the two-dimensional gel electrophoresis map of proteins in peripheral blood mononuclear cells (PBMCs) of healthy persons and RA patients (intervened or un-intervened with BZXD), analyzing the differential proteins and seeking out the RA associated proteins.

METHODSEighteen patients with early active RA were randomized into the BZXD group and the methotrexate (MTX) group, nine in each group, they were treated with BZXD (contained 15 Chinese herbs, as Herba Hedyotis diffusae, Herba Sarcandrae glabrae, Radix Salviae miltiorrhizae, Caulis Trachelosperi, Rhizoma Drynariae, Semen Coicis, etc.) and MTX combined with nimesulide Tablets respectively, three months as a treatment course, and their blood samples were collected for observation. Besides, blood samples from 9 healthy persons were taken as normal controls. PBMCs were isolated from blood using lymphozytes separation medium, and total protein in the cells was extracted through immobilized pH gradient two-dimensional gel electrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed using PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Then partial proteins were validated by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe 2-DE protein profile of PBMCs from healthy persons and RA patients before and 3 months after treatment were obtained, and 23 differential protein spots were found, 14 from 18 differential protein spots were successfully identified, of which 8 proteins were up-regulated and 6 proteins were down-regulated in RA patients as compared with control. After 3-month treatment, 5 differentially expressed proteins showed more obvious in the BZXD group than in the MTX group. RT-PCR verified that the expression of ApoA-I in all the three groups was consistent with the outcomes of 2-DE.

CONCLUSIONSSome differentially expressed proteins exist in the PBMCs of RA patients, which may play a potential role in the pathogenesis of RA; BZXD may treat RA by way of regulating the expression of some differential proteins in patients.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; drug therapy ; Blood Proteins ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Methotrexate ; therapeutic use ; Middle Aged ; Phytotherapy ; Proteome ; analysis ; Proteomics ; methods

OBJECTIVETo study the effect of Tongluo Xingnao effervescent tablets on learning and memory capacity and expression of Na(+)-K(+)-ATPase in hippocampus of rats with chronic cerebral ischemia-induced learning and memory dysfunction model.

METHODThe 2-VO method was used to establish sd rat model learning and memory dysfunction induced by chronic cerebral ischemia. The 50 rats in the successfully established model were randomly divided into the model control group, the Dihydroergotoxine Mesylate tablets group (0.7 mg x kg(-1), Tongluo Xingnao effervescent tablets high dose (7.56 g x kg(-1)), middle dose (3.78 g x kg(-1)) and low dose (1.59 g x kg(-1)) groups and the sham operation group (n = 10) as the control group. The groups were orally given 10 ml x kg(-1) x d(-1) drugs for consecutively 90 days. On the 86th day, Morris water maze was adopted for them. On the 90th day, a leaning and memory capacity test was held. The brain tissues were fixed with 10% formaldehyde and observed for pathomorphism after routine slide preparation and staining. The expression of hippocampal Na(+)-K(+)-ATPase was detected with immunohistochemistry and image quantitative analysis.

RESULTCompared with the model group, all of Tongluo Xingnao effervescent tablets groups showed significant decrease in the escape latency at the 5th day in the Morris water maze, and notable increase in the frequency of the first quadrant dwell, the frequency passing the escape platform and the frequency entering effective area (p < 0.05). According to the pathomorphological detection, the control group showed a significantly higher pathological score than the sham operation group (p < 0.01), the middle dose group showed a significantly lower pathological score than the model group (p < 0.05). According to the immunohistochemistical detection, the model control group showed a remarkably lower mean OD value of Na(+)-K(+)-ATPase than the sham operation group (p < 0.05), high and middle dose groups showed a significantly higher mean od value than the model control group (p < 0.01).

CONCLUSIONTongluo Xingnao effervescent tablets can improve the learning and memory capacity, reduce pathological changes of hippocampal tissues of rats with chronic cerebral ischemia-induced learning and memory dysfunction model, and promote the expression of Na(+)-K(+)-ATPase in hippocampus.

Animals ; Brain Ischemia ; drug therapy ; enzymology ; genetics ; psychology ; Chronic Disease ; drug therapy ; psychology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hippocampus ; drug effects ; enzymology ; Humans ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; genetics ; metabolism ; Tablets ; administration & dosage

Lactate dehydrogenase-C4 isozyme has been widely studied as the sperm-specific antigen in the research of the contraceptive vaccine. Developmental and testis-specific expression of the LDH-C gene requires mechanisms for activation in germ cells and repression in somatic cells. LDH-C promoter, several transcription factors and methylation of CpG dinucleotides have been proved of governing action on LDH-C gene transcription. The strategy is to induce antibodies in the female reproductive tract against LDH-C antigens at a sufficient level to block fertilization. Chemically modified LDH-C4 offers a potential application in the induction of infertility of homologous species in marked contrast to native LDH-C4. In addition, advances are being made in researches on the male contraceptive vaccine and synthetic peptide immunocontraceptive.
Animals ; Humans ; Isoenzymes ; genetics ; immunology ; L-Lactate Dehydrogenase ; genetics ; immunology ; Male ; Mice ; Papio ; Rabbits ; Spermatozoa ; enzymology ; Transcription, Genetic ; Vaccines, Contraceptive ; immunology

OBJECTIVETo understand the status of standard medication of antiretroviral therapy and identify its correlationship with preventing mother-to-child transmission of HIV (PMTCT).

METHODSQualitative and quantitative methods for measuring of standard medication towards antiretroviral therapy of PMTCT were used in a cross-sectional study of 167 pregnant women who had accessed to PMTCT services and 57 physicians providing service from January 2005 to June 2007 in 5 HIV epidemic counties in Yunnan province.

RESULTSA total of 167 mothers were recruited, of which 65.87% (110/167) demonstrated for standard medication. The percentage of rational selection of antivirus-therapy regimen was 88.02% (147/167). HIV positive mothers with rational using medicine during labor covered 81.37% (131/161). 87.42% (146/167) of women demonstrated good compliance. The main correlations with the standard medication level of HIV positive pregnant women and their infants were as follows: diagnose time (OR = 2.617; 95% CI: 1.184 - 5.783), place of delivery (OR = 0.064; 95% CI: 0.007 - 0.607), minorities (OR = 0.344; 95% CI: 0.162 - 0.730), understanding of HIV women for antiretroviral therapy (OR = 6.843; 95% CI: 1.449 - 32.312), and doctors' cognition about the regimen for antiretroviral therapy was not enough; 5 key knowledge points (purpose of PMTCT, the consequence of not requiring standard medication, the effect of knowing regimens, the relationship of CD4 levels and selected regimens, the side effect of the relevant medicine) score rate was 47.72% (136/285).

CONCLUSIONThe proportions of standard medication in PMTCT antiretroviral therapy were not high. Standard medication might be associated with multiple factors from doctors, patients and society. It is necessary to make some steps to improve standard medication of antiretroviral therapy in PMTCT.

Anti-HIV Agents ; therapeutic use ; China ; Cross-Sectional Studies ; Female ; HIV Infections ; drug therapy ; prevention & control ; transmission ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control ; Pregnancy ; Pregnancy Complications, Infectious ; drug therapy