Female ; Hemophilia A ; complications ; Humans ; Middle Aged ; Myasthenia Gravis ; complications

OBJECTIVETo investigate the effects of serotonin (5-HTIA) receptors in the hippocampal dentate gyrus (DG) on active avoidance learning in rats.

METHODSTotally 36 SD rats were randomly divided into control group, antagonist group and agonist group(n = 12). Active avoidance learning ability of rats was assessed by the shuttle box. The extracellular concentrations of 5-HT in the DG during active avoidance conditioned reflex were measured by microdialysis and high performance liquid chromatography (HPLC) techniques. Then the antagonist (WAY-100635) or agonist (8-OH-DPAT) of the 5-HT1A receptors were microinjected into the DG region, and the active avoidance learning was measured.

RESULTS(1) During the active avoidance learning, the concentration of 5-HT in the hippocampal DG was significantly increased in the extinction but not establishment in the conditioned reflex, which reached 164.90% ± 26.07% (P <0.05) of basal level. (2) The microinjection of WAY-100635 (an antagonist of 5-HT1A receptor) into the DG did not significantly affect the active avoidance learning. (3) The microinjection of 8-OH-DPAT(an agonist of 5-HT1A receptor) into the DG significantly facilitated the establishment process and inhibited the extinction process during active avoidance conditioned reflex.

CONCLUSIONThe data suggest that activation of 5-HT1A receptors in hipocampal DG may facilitate active avoidance learning and memory in rats.

8-Hydroxy-2-(di-n-propylamino)tetralin ; pharmacology ; Animals ; Avoidance Learning ; Dentate Gyrus ; physiology ; Piperazines ; pharmacology ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT1A ; physiology ; Serotonin ; physiology ; Serotonin Receptor Agonists ; pharmacology

OBJECTIVETo investigate the optimal extraction process of polysaccharides from S. fusiforme by enzymatic treatment.

METHODThe optimum extraction conditions were obtained by the experiment with the orthogonal design. The content of polysaccharides of S. fusiforme was determined by spectraphotometry.

RESULTThe amount of enzyme and temperature significantly affected total polysaccharides of S. fusiforme.

CONCLUSIONThe optimum extraction conditions include the addition of 1. 2 x 10 (4) U x 100 g(-1) enzyme into water at pH 4. 5, and the subsequent treatment for 10 min while the temperature is maintained at 45 degrees C.

Cellulase ; metabolism ; Hydrogen-Ion Concentration ; Polysaccharides ; analysis ; isolation & purification ; metabolism ; Sargassum ; chemistry ; Technology, Pharmaceutical ; methods ; Temperature

AIMTo study the feasibility of long-term potentiation(LTP) recording in the CA1 area of the rat in vivo with electrodes-binding technique.

METHODSAnesthetizing Wistar rats with urethane and fixing the animal on the stereotaxic device for acute surgery; implanting cannula into lateral cerebral ventricle; inserting self-made bound stimulating/recording electrodes into hippocampal CA1 area; recording basal field excitatory postsynaptic potential (fEPSP) and tetanus-induced long term potentiation (LTP).

RESULTSfEPSPs were reliably induced by using the stimulating/recording electrodes-binding technique, and the appearance rate of fEPSP was nearly 100%; basal fEPSP recording was very stable, lasting for long time enough to finish all experiment; high frequency stimulation (HFS) successfully induced LTP, which maintained more than three hours, the inductivity is about 67%; paired-pulse facilitation (PPF) recording was also stable; intracerebroventricular (i c v) injection of amyloid beta suppressed HFSinduced LTP evidently.

CONCLUSIONThe electrodes-binding technique for recording hippocampal LTP in vivo is quite simple and convenient. The experimental resource can be saved, and the rates of fEPSP appearance and LTP induction are kept high. Therefore, it is promising for this technique to be one electrophysiological auxiliary method in the research of learning and memory.

Animals ; Electric Stimulation ; methods ; Electrodes ; Excitatory Postsynaptic Potentials ; physiology ; Feasibility Studies ; Hippocampus ; physiology ; Long-Term Potentiation ; physiology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study the subcellular localization of severe fever with thrombocytopenia syndrome virus (SFTSV) in macrophages and understand the replication and assembly mechanism of SFTSV in host cells.

METHODSUsing two types of human macrophage cell lines THP-1 and U937, the study analyzed the intracellular colocalization of SFTSV with Golgi apparatus and endoplasmic reticulum by immunefluorescence staining and confocal microscopy.

RESULTSSFTSV infected macrophage cell lines THP-1 and U937. Immunofluorescence staining showed that the SFTSV nuclear protein colocalized with Golgi apparatus and closely surrounded by endoplasmic reticulum in the perinuclear region.

CONCLUSIONThe results suggested that Golgi complex and endoplasmic reticulum are probably the sites for formation and maturation of SFTSV viral particles.

Bunyaviridae ; isolation & purification ; Cell Line, Tumor ; Endoplasmic Reticulum ; virology ; Fever ; virology ; Golgi Apparatus ; virology ; Humans ; Macrophages ; virology ; Thrombocytopenia ; virology

BACKGROUNDMounting evidence suggests that tumors are histologically heterogeneous and are maintained by a small population of tumor cells termed cancer stem cells. CD133 has been identified as a candidate marker of cancer stem cells in laryngeal carcinoma. This study aimed to analyze the chemoresistance of CD133(+) cancer stem cells.

METHODSThe response of Hep-2 cells to different chemotherapeutic agents was investigated and the expression of CD133 was studied. Fluorescence-activated cell sorting analysis was used to identify CD133, and the CD133(+) subset of cells was separated and analyzed in colony formation assays, cell invasion assays, chemotherapy resistance studies, and analyzed for the expression of the drug resistance gene ABCG2.

RESULTSAbout 1% - 2% of Hep-2 cells were CD133(+) cells, and the CD133(+) proportion was enriched by chemotherapy. CD133(+) cancer stem cells exhibited higher potential for clonogenicity and invasion, and were more resistant to chemotherapy. This resistance was correlated with higher expression of ABCG2.

CONCLUSIONSThis study suggested that CD133(+) cancer stem cells are more resistant to chemotherapy. The expression of ABCG2 could be partially responsible for this. Targeting this small population of CD133(+) cancer stem cells could be a strategy to develop more effective treatments for laryngeal carcinoma.

AC133 Antigen ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Antigens, CD ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Carcinoma ; genetics ; metabolism ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Flow Cytometry ; Fluorouracil ; pharmacology ; Glycoproteins ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; metabolism ; Paclitaxel ; pharmacology ; Peptides ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Alveolar soft part sarcoma is a rare, aggressive malignancy of uncertain histological origin with a propensity for vascular invasion and distant metastasis. The case presented involves a 31-year-old woman with alveolar soft part sarcoma in the tongue root. The clinical features, pathogenesis, diagnosis and treatment were discussed.
Adult ; Female ; Humans ; Sarcoma, Alveolar Soft Part ; Tongue ; Tongue Neoplasms

OBJECTIVETo screen for the differential protein peaks of renal cell carcinoma (RCC) using magnetic beads-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

METHODSSerum proteins were profiled by magnetic beads (WCX) from 62 RCC patients and 37 patients with benign renal space-occupying lesions. Protein peaks were identified by MALDI-TOF-MS. Data were analyzed with Biomarker Wizard 3.1 and Biomarker Patterns Software 5.0. Diagnostic model for RCC was constructed based on 47 RCC cases and 26 patients with benign renal space-occupying lesions. The remaining 26 cases were evaluated with blind method.

RESULTSSeven differential protein peaks related to RCC were identified (Pβ0.05). The diagnostic model for RCC constructed by the differential protein peaks (m/z 2945.35, 15340.8, 6984.51, and 5819.23) generated excellent separation between the RCC and control groups, with a sensitivity of 83.0% and the specificity of 84.6%. As validated by blind method, the model had a sensitivity of 80.0% and a specificity of 81.8%.

CONCLUSIONDifferential protein peaks for RCC can be identified in serum by magnetic beads-based MALDI-TOF-MS, which is also valuable for the establishment of a RCC diagnostic model with a high sensitivity and specificity.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Carcinoma, Renal Cell ; blood ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Proteomics ; methods ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo establish decision tree and logistic regression classification models for diagnosing pancreatic adenocarcinoma (PaCa) and for screening serum biomarkers related to evaluation of different stages and curative effects.

METHODSSerum samples obtained from subjects with pancreatic adenocarcinoma (n = 58) and normal pancreas (n = 51) were applied to strong anion exchange chromatography (SAX2) chips for protein profiling by SELDI-TOF-MS to screen multiple serum biomarkers. Biomarker Wizard software and several statistical methods including algorithm of decision tree, logistic regression and ROC curves were used to construct the decision tree or logistic regression classification models.

RESULTSAverage of 61 mass peaks were detected at the molecular range of 2000-30,000, ten decision trees with the highest cross validation rate were chosen to construct the classification models, which can differentiate PaCa from normal pancreas with a sensitivity of 83.3% and a specificity of 100%. Logistic regression was used to achieve the AUC (0.976 +/- 0.011, P < 0.001) with a sensitivity of 77.6% - 91.4% and a specificity of 92.2% - 100%. Six mass peaks were combined by logistic regression to achieve the AUC 0.897 +/- 0.054, 0.978 +/- 0.021 and 0.792 +/- 0.107 (P < 0.05) in the three groups (patients at stage I and II, stage II and III, stage III and IV). One mass peak (M/Z 4,016) was screened (P < 0.05) significantly between the preoperative and postoperative PaCa samples and the intensity decreased weeks after operation.

CONCLUSIONDecision tree and logistic regression classification models of the mass peaks screened by SELDI-TOF-MS serum profiling can be used to differentiate pancreatic adenocarcinoma from normal pancreas, and is superior to CA 199. The detected mass peaks are helpful for the evaluation of curative effect and prognosis of pancreatic adenocarcinoma.

Adenocarcinoma ; blood ; diagnosis ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Area Under Curve ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Chromatography, Ion Exchange ; methods ; Decision Trees ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; blood ; diagnosis ; pathology ; surgery ; Prognosis ; Protein Array Analysis ; Proteomics ; ROC Curve ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics