Objective To investigate the exact protocol eliciting the hippocampal CA1 long-term depression (LTD) of rats in vivo and the effect of amyloid β-protein (Aβ) on the LTD.Method By applying test stimulation to Schaffer collateral in hippocampal CA1 region in rats,recorded the in vivo field excitatory postsynaptic potentials (fEPSPs) ;further,observed the induction of LTD with different low frequency stimulation (LFS) and investigated the effect of Aβ25-35 on the LTD.Results Prolonged LFS (1,5 and 10 Hz) but not paired-pulse stimulus (PPS) effectively elicited the LTD in the hippocampal CA1 region,with significantly decreased amplitude of fEPSPs after LFS ; 1 Hz 900 pulses group induced a stronger LTD,being (63.7 ± 3.8) ％ at 120 min post-LFS,lower (P ＜ 0.05) than (75.1 ± 3.2) ％ in 600 pulses group ; different frequencies (1,5 and 10 Hz) of LFS with same pulses induced similar degree of LTD,the amplitude of fEPSPs were (63.7 ± 3.8) ％,(61.2 ± 3.6) ％ and (59.8 ± 3.9) ％ respectively,without significant differences between any two groups (P ＞ 0.05) ; after applying 12.5 nmol and 25 nmol Aβ25-35,the amplitude of fEPSPs decreased to (63.2 ± 3.8) ％ and (46.8 ± 3.9) ％,respectively,and lower and than that in control ((73.9 ± 3.0) ％,P ＜ 0.05).Conclusion Prolonged LFS effectively induced in vivo hippocampal LTD of rats,which provides an important electrophysiological protocol for the study of synaptic plasticity; Aβ25-35 injection dont affect the baseline synaptic transmission,but dose-dependently enhance the in vivo hippocampal LTD of rats,indicating that Aβ-induced LTD facilitation may be involved the early impairment of learning and memory in Alzheimer's disease.

Objective To study the relationship between asthma and mRNA expression of β_2AR and IL-12 gene;to determine the expression level of β_2AR, IL-12 related to CysLT_1-receptor;to analyze the correlation between the severity of asthma and the level of the genes expression. Methods White blood cells was drawn from peripheral blood mononuelear cells of asthmatic children. DNA was taken and related genes were analyzed for half quantitative analysis by D-congeal glue analyzer software. Results The level of IL-12. β_2AR mRNA expression in the asthma group is obviously lower than healthy controls (P < 0.01) ;when asthma attacks, the expression of IL-12 is positively correlated to that of β_2AR (r = 0.34, P = 0.001) and negatively correlated to that of CysLT_1-receptor (r = -0.92, P = 0.001), the expression of β_2AR is negatively correlated to the expression of CysLT_1-receptor (r = -0.85, P = 0.003). The express of IL-12 is not related to the severity of asthma (P = 0.16), while there was a negative correlation between the expression of β_2AR and the severity of asthma (P = 0.003). Conclusions The expression level of IL-12, β_22AR in the asthmatic children is obviously lower than normal;The expression level of IL-12 is positively correlated to β_2AR in PBMC of asthmatic children;The expression level of CysLT_1-receptor is negatively correlated to IL-12 and β_2AR in the PBMC of asthmatic children;The expression of β_2AR is negatively correlated to the severity of asthma.

Objective To study effects of yinlingⅠon cell viability and oxidative injury of lead-exposed cell.Methods After lead exposure Vero cell was treated with yinlingⅠof different concentrations.The cell viablitity was measured by methyl thiaxiolyl tetrazolium(MTT) method and the superoxide dismutase(SOD)and malondialdehyde(MDA) activity in cell suspensions were measured.Results When Vero cell lived in 125 ?mol/L lead acetate surrounding,the MDA concentration increased,but the viability of cell and the SOD content in the Vero cell suspension decreased.YinlingⅠcould increase the viability of lead-exposed cell during a certain extent;in the most non-toxicity concentration yinlingⅠcould elevate the SOD content in the Vero cell suspension,reduced the MDA concentration and resisting lead toxication in vitro.Conclusion YinlingⅠhas the protective effects on the cell viability and oxidative injury of lead-exposed cell.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Objective To discuss real time three-dimensional reconstruction techniques of spiral CT in the detection of tracheoesophageal groove lymph node metastasis and its diagnostic value.Methods Spiral CT RT3D reconstruction of the supraclaviculrar region were applied to 45 patients with scanning and post-processing on the workstation.Mediastinum,color in chest,clip planes 3D images were obtained by adjusting the CT value threshold and encoding pseudo color techniques.Results RT3D images can correctly reveal the macroscopic morphology of lymphatic metastasis and it's pressure to the trachea and carotid artery.There were significant differennces of visualization capabilities of radiological signs among different 3D techniques.Conclusion With the utilization of multiple methods,RT3D reconstruction can play an important role in detecting tracheoesophageal groove lymph node metastasis of esophageal cancer.It is an important complement to the axial CT images and the basis of clinical treatment.

Objective To investigate the correlation between the quantitative results,the semi-quantitative scores from ultrasound elastography and the distribution of myofibroblasts (MFS)in breast tumor,and to analyze the value of quantitative results and the semi-quantitative score from ultrasound elastography in the diagnosis of breast tumor.Methods Thirty eight patients with breast lesions underwent ultrasound elasticity examinations,tissue dispersion quantitative analysis technique was used to assess the 11 characteristic quantities and the corresponding strain ratios in all lesions,and the score of ultrasonic elastography was evaluated.38 cases were divided into benign and malignant group according to pathological diagnosis results.The expression levels of CD34 andα-SMA protein in breast tissue were examined by immunohistochemistry.The distribution patterns of MFS in breast tumor were analyzed. The correlation between the quantitative results, the semi-quantitative scores from ultrasound elastography and the distribution of MFS in breast tumor was studied.Results The expression level of CD34 in malignant group was significantly higher than that in benign group, while the expression level of α-SMA was significantly higher than that in benign group;the differences in the expression levels of CD34 andα-SMA between benign and malignant breast tumor patients were statistically significant (P < 0.01 ).The masses observed in malignant group by elastography were shown in blue,while most of the masses in benign group were shown in green.There were statistically significant differences between two groups in elastography scores (P < 0.05 ). There was a anegative correlation between the CD34 expression in tumor tissue and the score of ultrasound elastography (r=-0.423 7,P =0.027 3).There was a negative correlation between the CD34 expression and the score of ultrasound elastography (r=-0.423 7,P =0.027 3),while the positive correlation was found between theα-SMA expression and the score of ultrasound elastography (r=0.397 0,P =0.014 2).The CD34 expression was significantly correlated with the average relative strain value,entropy,area ratio of low-strain region,kurtosis, skewness and inverse difference moment (P <0.05).Among the 11 characteristics,CD34 was positively correlated with the average relative strain value (r=0.385 6,P =0.016 8)and entropy (r=0.380 5,P =0.018 5);CD34 was negatively correlated with area ratio of low-strain region (r = - 0.351 7,P = 0.030 4),kurtosis (r =-0.427 7,P =0.007 4),skewness (r=-0.394 6,P =0.014 2),inverse difference moment (r = -0.344 3, P =0.034 3),angular second moment (r = - 0.484 9,P = 0.002 0)and strain ration (r = - 0.379 0,P =0.047 5);CD34 was not correlated with standard deviation,complexity and contrast (P > 0.05).The α-SMA espression was positively correlated with kurtosis (r = 0.356 9,P = 0.027 8),skewness (r = 0.323 0,P =0.047 9),area ratio of low-strain region (r=0.382 0,P =0.021 6)and strain ratio (r=0.403 3,P =0.012 0). Conclusion The features and its scores of ultrasound elastography are correlated with the distribution of MFS in breast tumor,suggesting that the ultrasound elastography is very informative and helpful in the diagnosis of breast tumor.

The role of flavonoids of Echinps latifolius (FELT) in Wnt signaling was investigated in adjuvant arthritis (AA) rats. The therapeutic effects of FELT on AA rats were detected by rat arthritis score and MTT. The effect of FELT gavage treatment on the Wnt signaling key gene β-catenin, C-myc and cyclin D1 in synovium from AA rats was detected by Real-time qPCR, and the effects of FELT gavage treatment on the upstream negative regulation gene SFRP 1,2,4,5 in synovium from AA rats were detected by Real-time qPCR. The results showed that FELT gavage treatment significantly inhibited arthritis score and MTT values in AA rats, significantly inhibited the expression of the Wnt signaling gene β-catenin, C-myc and cyclin D1, significantly up-regulated the expression of the up- stream negative regulation gene SFRP 1,2,4. FELT has a better therapeutic effect for AA rats.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Asteraceae ; chemistry ; Disease Models, Animal ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Flavonoids ; administration & dosage ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Synovial Membrane ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of epirubicin hydrochloride (EPI) in rat plasma, daunorubicin hydrochloride was used as internal standard. The plasma samples were deproteinated with methanol, and separation was performed on a reversed-phase CAPCELL PAK C18 column (3.0 mm x 50 mm, 3 microm). The mobile phase contained methanol-0.1% formic acid (80:20). Detection was carried out by multiple reaction monitoring on a HP1200-6410 QQQ LC/MS system. Different preparations of EPI solution, EPI-LIP (EPI-liposome) and EPI-LTSL (EPI-thermosensitive liposome) was administered in rats by i.v with the same dosage (12 mg kg(-1)). The pharmacokinetic model and parameters were fitted and calculated by the DAS ver2.0 software. The calibration curve was linear in the range of 0.01-50 microg mL(-1). The limit of quantification was 0.01 microg mL(-1). RSDs of intra- and interbatch precisions were all less than 11.9%. The average extract recovery was 89.3% and 92.1%, respectively. The pharmacokinetics of EPI in rats with all preparations were fitted to three compartments, which all fast distributed and slowly eliminated. The t1/2 alpha, t1/2 beta, t1/2 gamma, AUC(0-infinity), and MRT(0-infinity) of EPI-LTSL group were 7.5, 1.3, 12.6, 12.9, 3.7 times those of EPI solution group; and 1.6, 1.4, 12.3, 2.9, 2.6 times those of EPI-LIP group. Moreover, the CL of the latter two groups was about 13.4 times of the former EPI-LTSL group. EPI-LTSL can significantly improve AUC and prolong the circulation time of EPI in rat plasma.
Animals ; Antibiotics, Antineoplastic ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, Liquid ; Drug Carriers ; Epirubicin ; administration & dosage ; blood ; pharmacokinetics ; Liposomes ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Tandem Mass Spectrometry

To study the effect of pulchinenoside (PULC) on the Frizzled (FZD) expression of adjuvant arthritis ( AA) rats. AA rats were prepared through the toe injection with complete Freund's adjuvant to culture fibroblast-like synoviocytes (FLS). The effect of the oral administration with PULC on the FZD8 expression was detected by the real time qPCR. The effect of FZD8 knockout on the expressions of IL-1, IL-6, IL-8 were detected by MTT and ELISA. The role of miR-375 in the abnomal expression of FZD8 was detected by the real time qPCR. The results showed signfiicant decrease in the FZD8 expression among AA rats, FLS proliferation ater FZD8 knockout and IL-1, IL-6, IL-8 expressions and notable increase in miR-375 expression after the oral administration with PULC. The up-regulated miR-375 expression can inhibit the FZD8 expression. PULC may inhibit the FZD8 expression by up-regulating the miR-375 expression.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism ; Saponins ; administration & dosage

The role of pulchinenoside (PULC) in the regulation of MeCP2 expression was investigated in RA model rats. Adjuvant arthritis rats were used as RA model rats, and fibroblast-like synoviocytes (FLS) from the RA model rats were cultured. The effect of 100 mg x kg(-1) PULC gavage treatment on the MeCP2 expression and the effect of MeCP2 siRNA on the expression of SFRP2 and β-catenin were detected by real time qPCR and Western blotting. The role of PULC in the FLS proliferation was detected by MTT. The results showed that the MeCP2 expression was down-regulated, the SFRP2 expression was up-regulated and the FLS proliferation was inhibited in FLS after therapy. MeCP2 siRNA significantly inhibited the MeCP2 expression, up-regulated the SFRP2 expression and inhibited the β-catenin expression in FLS from RA model rats. PULC may increase the SFRP2 expression, inhibit the Wnt signaling and inhibit the FLS proliferation in FLS from the RA model rats by inhibiting the MeCP2 expression.
Animals ; Arthritis, Rheumatoid ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Fibroblasts ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Methyl-CpG-Binding Protein 2 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; cytology ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; genetics ; metabolism